Datasets:

bigbio

/

medhop

like 1

MH_train_1100

MH_train_1100

MH_train_1100

interacts_with DB00163?

multiple_choice

Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . 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 . 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 . Effects of retroviral-mediated P08183 expression on hematopoietic stem cell self-renewal and differentiation in culture . Ex vivo expansion of hematopoietic stem cells would be useful for bone marrow transplantation and gene therapy applications . Toward this goal , we have investigated whether retrovirally-transduced murine stem cells could be expanded in culture with hematopoietic cytokines . Bone marrow cells were transduced with retroviral vectors expressing either the human multidrug resistance 1 gene ( HaMDR1 ) , a variant of human dihydrofolate reductase ( HaDHFR ) , or both P08183 and P00374 in an internal ribosomal entry site ( IRES ) -containing bicistronic vector ( HaMID ) . Cells were then expanded for 15 days in cultures stimulated with interleukin ( IL ) -3 , P05231 , and stem cell factor . When very low marrow volumes were injected into lethally irradiated recipient mice , long-term reconstitution with 100 % donor cells was seen in all mice injected with HaMDR1- or HaMID-transduced cells . By contrast , engraftment with HaDHFR- or mock-transduced cells ranged from partial to undetectable despite injection of significantly larger marrow volumes . In addition , mice transplanted with expanded HaMDR1- or HaMID-transduced stem cells developed a myeloproliferative disorder that was characterized by an increase in abnormal peripheral blood leukocytes . These results show that P08183 -transduced stem cells can be expanded in vitro with hematopoietic cytokines , but indicate that an increased stem cell division frequency can lead to stem cell damage . Estrogen regulation in human breast cancer cells of new downstream gene targets involved in estrogen metabolism , cell proliferation and cell transformation . We explored , by cDNA mini-arrays , gene expression measurements of MVLN , a human breast carcinoma cell line derived from MCF-7 , after 4 days of exposure to 17beta-estradiol ( E(2) ) treatment , in order to extend our understanding of the mechanism of the pharmacological action of estrogens . We focused on 22 genes involved in estrogen metabolism , cell proliferation regulation and cell transformation . The specificity of the E(2) response was reinforced by comparison with 4-hydroxytamoxifen ( OH-Tam ) , DB00947 and E(2)+OH-Tam expression profiles . Real-time quantitative PCR ( RTQ-PCR ) confirmed the variation of expression of known ( P04155 , P15514 , P35568 , P22692 , P12004 , P04626 , P07339 , MYC ) as well as novel ( DLEU2 , P20248 , P22309 , O15438 , O15440 , O75410 , P20827 , NOV , P01040 , P51511 , O75362 ) genes . The temporal response of these gene expression regulations was then investigated after 6 and 18 h of E(2) treatment and this allowed the identification of different time-course patterns . Cycloheximide treatment studies indicated first that estrogen affected the transcript levels of O15438 and O15440 through dissimilar pathways , and secondly that protein synthesis was needed for modulation of the expression of the P20248 and O75410 genes by estrogens . Western blot analysis performed on P04155 , P35568 , P22692 , amphiregulin , P12004 , cyclin A2 , O75410 and O15440 proteins confirmed the mini-array and RTQ-PCR data , even for genes harboring low variations of mRNA expression . Our findings should enhance the understanding of changes induced by E(2) on the transcriptional program of human E(2)-responsive cells and permit the identification of new potential diagnostic/prognostic tools for the monitoring of estrogen-related disease conditions such as breast cancer . Interaction between P20292 and P20815 gene variants significantly increases the risk for cerebral infarctions in Chinese . In this study , we investigated associations between susceptibility genes and cerebral infarctions in a Chinese population , and whether gene-gene interactions increase the risk of cerebral infarctions . Overall , 292 patients with cerebral infarctions and 259 healthy control individuals were included . Eight variants in five candidate genes were examined for the risk of stroke , including the SG13S32 ( rs9551963 ) , SG13S42 ( rs4769060 ) , SG13S89 ( rs4769874 ) , and SG13S114 ( rs10507391 ) variants of the P09917 activating protein ( P20292 ) gene , the G860A ( rs751141 ) variant of the soluble epoxide hydrolase ( P34913 ) gene , the A1075C ( rs1057910 ) variant of the P11712 *2 gene , the C430T ( rs1799853 ) variant of the P11712 *3 gene , and the A6986G ( rs776746 ) variant of the P20815 gene . Gene-gene interactions were explored using generalized multifactor dimensionality reduction methods . There were no statistically significant differences in the frequencies of the genotypes of the eight candidate genes . The generalized multifactor dimensionality reduction analysis showed a significant gene-gene interaction between SG13S114 and A6986G , with scores of 10 for cross-validation consistency and 9 for the sign test ( P=0.0107 ) . These gene-gene interactions predicted a significantly higher risk of cerebral infarction ( adjusted for age , hypertension , and diabetes mellitus ; odds ratio=1.80495 % , confidence interval : 1.180-2.759 , P=0.006 ) . A two-loci gene interaction confers a significantly higher risk for cerebral infarction . The combinational analysis used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases . 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 . 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 . 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 . Evaluation of synthetic polymeric micelles as a stabilization medium for the handling of membrane proteins in pharmaceutical drug discovery . PURPOSE : Polymeric micelles have been used for solubilization of insoluble drugs and as carriers for drug delivery applications . Here we evaluated an application of the synthetic polymeric micelles in experiments designed to improve the handling and stability of membrane proteins targets . METHODS : Particle sizing by dynamic light scattering was performed in a Zeta Plus Photon Correlation Spectrometer at 532 nm . P22309 activity has been measured in fluorescent assay using scopoletin as a substrate . P35354 activity has been measured in a fluorescent assay using Amplex Red . Fluorescence Resonance Energy Transfer ( FRET ) was monitored using either 463 nm excitation wavelength ( the emission range 500-600 nm ) or 395 nm excitation wavelength ( the emission range 500-600 nm ) . RESULTS : Incorporation of membrane proteins into PreserveX-QML polymeric micelles resulted in improved homogeneity and stability of the preparation and in reduced light scattering . Stabilization of the biological activity of micelle-incorporated membrane proteins , such as the human P22309 and P35354 both during extended incubations at room temperature and during multiple freeze/thaw cycles , has been achieved . CONCLUSION : PreserveX-QML polymeric micelles help to homogenize and disperse membrane proteins preparations and stabilize the biological activity of the proteins making it more suitable for pharmaceutical assays and applications . Novel target for induction of apoptosis by cyclo-oxygenase-2 inhibitor SC-236 through a protein kinase C-beta(1)-dependent pathway . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) reduce the risk of gastrointestinal cancers . Recently , a similar protective effect has been demonstrated by the specific cyclo-oxygenase-2 ( P35354 ) inhibitors . However , the exact mechanism that accounts for the anti-proliferative effect of specific P35354 inhibitors is still not fully understood , and it is still controversial whether these protective effects are predominantly mediated through the inhibition of P35354 activity and prostaglandin synthesis . Identification of molecular targets regulated by P35354 inhibitors could lead to a better understanding of their pro-apoptotic and anti-neoplastic activities . In the present study , we investigated the effect and the possible molecular target of a P35354 -specific inhibitor SC-236 on gastric cancer . We showed that SC-236 induced apoptosis in gastric cancer cells . However , this effect was not dependent on P35354 inhibition . SC-236 down-regulated the protein expression and kinase activity of P05771 (1) , increased the expression of PKCdelta and PKCeta , but did not alter the expression of other PKC isoforms in AGS cells . Moreover , exogenous prostaglandins or PGE(2) receptor antagonists could not reverse the inhibition effect on PKCbeta(1) by SC-236 , which suggested that this effect occurred through a mechanism independent of cyclo-oxygenase activity and prostaglandin synthesis . Overexpression of PKCbeta(1) attenuated the apoptotic response of AGS cells to SC-236 and was associated with overexpression of P38936 (waf1/cip1) . Inhibition of PKCbeta(1)-mediated overexpression of P38936 (waf1/cip1) partially reduced the anti-apoptotic effect of PKCbeta(1) . The down-regulation of PKCbeta(1) provides an explanation for P36551 -independent apoptotic effects of specific P35354 inhibitor in cultured gastric cancer cells . We also suggest that PKCbeta(1) act as survival mediator in gastric cancer , and its down-regulation by P35354 inhibitor SC-236 may provide new target for future treatment of gastric cancer . 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 . 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 . 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 . 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 . Large-scale association study for structural soundness and leg locomotion traits in the pig . BACKGROUND : Identification and culling of replacement gilts with poor skeletal conformation and feet and leg ( FL ) unsoundness is an approach used to reduce sow culling and mortality rates in breeding stock . Few candidate genes related to soundness traits have been identified in the pig . METHODS : In this study , 2066 commercial females were scored for 17 traits describing body conformation and FL structure , and were used for association analyses . Genotyping of 121 SNPs derived from 95 genes was implemented using Sequenom 's MassARRAY system . RESULTS : Based on the association results from single trait and principal components using mixed linear model analyses and false discovery rate testing , it was observed that P02649 , P34820 , P30988 , P08123 , P20849 , DKFZ , P35555 and VDBP were very highly significantly ( P < 0.001 ) associated with body conformation traits . The genes P09917 , P34820 , P30988 , O00300 , P30559 and Q9UBV4 were very highly significantly ( P < 0.001 ) associated with FL structures , and P02649 , P30988 , P08123 , P30968 , Q14623 , P42898 and Q9UBV4 were highly significantly ( P < 0.01 ) associated with overall leg action . Strong linkage disequilibrium between P30988 and P08123 on SSC9 was detected , and haplotype -ACGACC- was highly significantly ( P < 0.01 ) associated with overall leg action and several important FL soundness traits . CONCLUSION : The present findings provide a comprehensive list of candidate genes for further use in fine mapping and biological functional analyses . P10275 signals regulate UDP-glucuronosyltransferases in the urinary bladder : a potential mechanism of androgen-induced bladder carcinogenesis . UDP-glucuronosyltransferases ( UGTs ) , major phase II drug metabolism enzymes , play an important role in urinary bladder cancer initiation by detoxifying carcinogens . We aimed to determine if androgens regulate P78381 expression via the androgen receptor ( AR ) pathway in the bladder . Real-time reverse transcription-polymerase chain reaction and Western blot analyses were used to assess P22309 levels in the normal urothelium SVHUC cell line stably expressed with AR and in bladder tissues from AR knockout ( ARKO ) and castrated male mice . Immunohistochemistry was also performed in radical cystectomy specimens . DB02901 ( DB02901 ) treatment in SVHUC-AR reduced mRNA expression of all the P22309 subtypes ( 19-75 % decrease ) , and hydroxyflutamide antagonized the DB02901 effects . In contrast , DB02901 showed only marginal effects on P22309 expression in SVHUC-Vector . Of note were higher expression levels of UGT1As in SVHUC-Vector than in SVHUC-AR . In ARKO mice , all the Ugt1a subtypes were up-regulated , compared to wild-type littermates . In wild-type male mice , castration increased the expression of Ugt1a8 , Ugt1a9 , and Ugt1a10 . Additionally , wild-type female mice had higher levels of Ugt1a than wild-type males . Immunohistochemical studies showed strong ( 3+ ) P22309 staining in 11/24 ( 46 % ) cancer tissues , which was significantly lower than in corresponding benign tissues [ 17/18 ( 94 % ) cases ( P = 0.0009 ) ] . These results suggest that androgen-mediated AR signals promote bladder carcinogenesis by down-regulating the expression of UGTs in the bladder . 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 . Q16552 promotes p38 MAPK-dependent endothelial activation enhancing neutrophil recruitment to sites of inflammation . Neutrophilic inflammation plays an important role in lung tissue destruction occurring in many chronic pulmonary diseases . Neutrophils can be recruited to sites of inflammation via the action of the cytokine Q16552 . In this study , we report that Q96F46 and Q8NAC3 mRNA expression is significantly increased in asthmatic bronchoscopic biopsies and that these receptors are not only expressed on epithelial and inflammatory cells but also on endothelial cells . Q16552 potently stimulates lung microvascular endothelial cells to produce chemoattractants ( P10145 and derivatives of the P09917 pathway ) that selectively drive neutrophil but not lymphocyte chemotaxis . Moreover , Q16552 promotes endothelial activation by inducing the expression of endothelial adhesion markers ( P16581 , P19320 , and P05362 ) in a p38 MAPK-dependent manner . This increased expression of adhesion molecules stimulates the trans-endothelial migration of neutrophils , as well as the transmigration of HT-29 colon carcinoma cells , suggesting a further role in promoting lung metastasis . Finally , Q16552 increased neutrophil adhesion to the endothelium in vivo as determined by intravital microscopy of mice cremaster muscle . Overall , our results demonstrate that Q16552 is a potent activator of the endothelium in vivo leading to neutrophil infiltration . Therefore , preventing neutrophil recruitment by blocking the action of Q16552 on endothelial cells may prove to be highly beneficial in diseases in which neutrophilic inflammation plays a key role . Activation of natural interferon-alpha producing cells by apoptotic U937 cells combined with lupus IgG and its regulation by cytokines . We recently demonstrated that IgG from patients with systemic lupus erythematosus ( SLE ) in combination with U937 cells made apoptotic by UV-irradiation , can induce interferon-alpha ( IFN-alpha ) production in normal peripheral blood mononuclear cells ( PBMC ) . In the present study we show by flow cytometry that the actual IFN-alpha producing cells ( IPC ) among PBMC had the same phenotype ( HLA-DR+ , P01730 + , CD11b- , CD11c- , P08571 - , P15391 - , CD32- , P16671 + , P25942 + , CD45RA+ , P34810 + , Q01151 + , P42081 - , IL-3R+ and IL-10R- ) and low frequency ( approximately 2/10(4)PBMC ) as the IPC activated by Herpes simplex virus type I . Consequently , these cells correspond to the natural IPC , also described as type 2 precursor dendritic cells . We also demonstrated that cytokines of possible importance in the pathogenesis in SLE had effects on the IFN-alpha production . Specifically , the IFN-alpha production was strongly increased by the type I IFNs , IFN-alpha and -beta , but markedly inhibited by P22301 and also to some extent by TFN-alpha . In contrast , the cytokines P01579 , P05231 , TGF-beta and GM- P04141 had no clear effects . No production of P22301 was detected in PBMC stimulated by apoptotic U937 cells and SLE IgG . These results may explain the cause of the ongoing IFN-alpha production in SLE patients and its relation to the autoimmune process . 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 . 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 . Cytokine-induced bronchoconstriction in precision-cut lung slices is dependent upon cyclooxygenase-2 and thromboxane receptor activation . Cytokines play an essential role in the regulation of inflammatory responses . The effects of cytokines on lung functions are less well known and their study in vivo is complicated by the attraction of leukocytes to the inflamed sites . Recently the model of precision-cut lung slices was developed , where viable lung slices with an intact microanatomy are taken into culture and where bronchoconstriction can be followed by observing single airways under the microscope . We used this model to study the direct effects of cytokines on airway tonus in the absence of blood-derived leukocytes . Incubation of precision-cut lung slices with a mixture of tumor necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -1beta , and interferon ( IFN ) -gamma resulted in contraction of airways , which was accompanied by expression of cyclooxygenase ( Cox ) -2 and thromboxane release into the supernatant . The thromboxane receptor antagonist SQ29548 completely prevented the cytokine-induced bronchoconstriction , whereas the P09917 inhibitor AA681 had no effect on cytokine-induced bronchoconstriction . Preventing the expression of Cox-2 by dexamethasone or blocking Cox-2 activity with the selective Cox-2 inhibitor NS398 attenuated both thromboxane formation and bronchoconstriction . Incubation of lung slices with each of the cytokines alone caused no bronchoconstriction ; in fact , IL-1 alone rather dilated the airways . However , simultaneous incubation with P01375 and IL-1beta caused a bronchoconstriction that was not further enhanced by P01579 . We conclude that P01375 and IL-1beta synergistically cause bronchoconstriction by induction of Cox-2 and subsequent activation of the thromboxane receptor . Our study raises the possibility that P01375 and IL-1 may contribute to bronchospasm during inflammatory lung diseases . P01375 -induced cyclooxygenase-2 expression in human lung epithelial cells : involvement of the phospholipase C-gamma 2 , protein kinase C-alpha , tyrosine kinase , NF-kappa B-inducing kinase , and I-kappa B kinase 1/2 pathway . P01375 induced a dose- and time-dependent increase in cyclooxygenase-2 ( P35354 ) expression and DB00917 formation in human NCI-H292 epithelial cells . Immunofluorescence staining demonstrated that P35354 was expressed in cytosol and nuclear envelope . Tyrosine kinase inhibitors ( genistein or herbimycin ) or phosphoinositide-specific phospholipase C inhibitor ( U73122 ) blocked P01375 -induced P35354 expression . P01375 also stimulated phosphatidylinositol hydrolysis and protein kinase C ( PKC ) activity , and both were abolished by genistein or U73122 . The PKC inhibitor , staurosporine , also inhibited P01375 -induced response . The 12-O-tetradecanoylphorbol 13-acetate ( TPA ) , a PKC activator , also stimulated P35354 expression , this effect being inhibited by genistein or herbimycin . NF-kappaB DNA-protein binding and P35354 promoter activity were enhanced by P01375 , and these effects were inhibited by genistein , U73122 , staurosporine , or pyrolidine dithiocarbamate . TPA stimulated both NF-kappaB DNA-protein binding and P35354 promoter activity , these effects being inhibited by genistein , herbimycin , or pyrolidine dithiocarbamate . The P01375 -induced , but not the TPA-induced , P35354 promoter activity was inhibited by phospholipase C-gamma2 mutants , and the P35354 promoter activity induced by either agent was attenuated by dominant-negative mutants of P17252 , NF-kappaB-inducing kinase , or I-kappaB ( inhibitory protein that dissociates from NF-kappaB ) kinase (IKK)1 or 2 . IKK activity was stimulated by both P01375 and TPA , and these effects were inhibited by staurosporine or herbimycin . These results suggest that , in NCI-H292 epithelial cells , P01375 might activate phospholipase C-gamma2 via an upstream tyrosine kinase to induce activation of P17252 and protein tyrosine kinase , resulting in the activation of NF-kappaB-inducing kinase and O15111 /2 , and NF-kappaB in the P35354 promoter , then initiation of P35354 expression and DB00917 release . 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 . 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 . Association of polymorphisms in P36888 , P00533 , P09917 , and Q8TAT5 with glioblastoma in the Han Chinese population . Glioblastoma ( GBM ) is the highest-grade glioma in astrocytoma . Patients often have poor prognosis due to therapeutic resistance and tumor recurrence . Identification of the genetic factors of GBM could be important contribution to early prevention of this disease . We genotyped 17 tag single-nucleotide polymorphisms ( tSNPs ) from nine genes in this study , including 72 cases and 302 controls . SNP genotyping was conducted using Sequenom MassARRAY RS1000 . Statistical analysis of the association between tSNPs and GBM was performed using the χ ( 2 ) test and SNPStats software . The rs3829382 in P36888 was associated with increased odds of developing GBM using the χ ( 2 ) test . When we analyzed tSNPs under different inheritance models , we found rs9642393 in P00533 increased odds of developing GBM in the dominant model . After stratification by gender , we found that rs12645561 in Q8TAT5 and rs2291427 in P09917 were associated with developing GBM . Polymorphisms within P36888 , P00533 , Q8TAT5 , and P09917 may contribute to the occurrence of GBM in the Han Chinese population . However , the functional significance of these polymorphisms needs further investigation . DB00163 , P09917 and oxidative stress in haemodialysis patients : facts , not fancies . 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 . 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 . 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 . 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 . PKC-beta1 mediates glucose-induced Akt activation and TGF-beta1 upregulation in mesangial cells . Accumulation of glomerular matrix is a hallmark of diabetic nephropathy . The serine/threonine kinase Akt mediates glucose-induced upregulation of collagen I in mesangial cells through transactivation of the P01133 receptor ( P00533 ) . In addition , in renal tubular cells , glucose-induced secretion of TGF-beta requires phosphoinositide-3-OH kinase , suggesting a possible role for Akt in the modulation of TGF-beta expression , but the mechanisms of Akt activation and its involvement in TGF-beta regulation are unknown . Here , in primary mesangial cells , high glucose induced AktS473 phosphorylation , which correlates with its activation , in a protein kinase C beta ( P05771 ) -dependent manner . DB09341 led to PKC-beta1 membrane translocation and association with Akt , and PKC-beta1 immunoprecipitated from glucose-treated cells phosphorylated recombinant Akt on S473 . PKC is known to mediate glucose-induced TGF-beta1 upregulation through the transcription factor AP-1 ; here , inhibitors of phosphoinositide-3-OH kinase , P05771 and Akt , and dominant-negative Akt all prevented glucose-induced activation of AP-1 and upregulation of TGF-beta1 . Finally , pharmacologic and dominant negative inhibition of P00533 blocked glucose-induced activation of PKC-beta1 , phosphorylation of AktS473 , activation of AP-1 , and upregulation of TGF-beta1 . In vivo , the P05771 inhibitor ruboxistaurin prevented Akt activation in the renal cortex of diabetic rats . In conclusion , PKC-beta1 is an Akt S473 kinase in glucose-treated mesangial cells , and TGF-beta1 transcriptional upregulation requires P00533 /PKC-beta1/Akt signaling . New therapeutic approaches for diabetic nephropathy may result from targeting components of this pathway , particularly the initial P00533 transactivation . 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 . 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 . 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 . Fatal rhabdomyolysis in a patient with liver cirrhosis after switching from simvastatin to fluvastatin . P04035 inhibitors ( statins ) are widely used to treat hypercholesterolemia . Among the adverse effects associated with these drugs are statin-associated myopathies , ranging from asymptomatic elevation of serum creatine kinase to fatal rhabdomyolysis . DB01095 -induced fatal rhabdomyolysis has not been previously reported . We describe here a patient with liver cirrhosis who experienced fluvastatin-induced fatal rhabdomyolysis . This patient had been treated with simvastatin ( 20 mg/day ) for coronary artery disease and was switched to fluvastatin ( 20 mg/day ) 10 days before admission . He was also taking aspirin , betaxolol , candesartan , lactulose , and entecavir . Rhabdomyolysis was complicated and continued to progress . He was treated with massive hydration , urine alkalization , intravenous furosemide , and continuous renal replacement therapy for acute renal failure , but eventually died due to rhabdomyolysis complicated by hepatic failure . In conclusion , fluvastatin should be used with caution in patients with liver cirrhosis , especially with other medications metabolized with P11712 . 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 . 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 . 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 . 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 . 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 ) . 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 . 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 . 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 . 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 . 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 . DB00163 80th anniversary : a double life , not only fighting radicals . Recent research on DB00163 has revealed specific cellular functions of this compound belonging to the vitamin E family . Alpha-tocopherol can act as a radical scavenger , as a pro-oxidant , as an anti-alkylation agent and , most important , by mechanisms that are independent of the above properties . To the last group belong protein kinase C and P09917 inhibition at post-translational level , as well as DB00163 activation of protein phosphatase 2A and diacylglycerol kinase . Furthermore , at transcriptional level , several genes ( P16671 , alpha-TTP , alpha-tropomyosin , and collagenase ) are modulated by DB00163 . These effects result in inhibition of smooth muscle cell proliferation , platelet aggregation , and monocyte adhesion and may be related to the alleged protection of atherosclerosis by vitamin E. On the other side , epidemiological and intervention studies have shown some inconsistent results . Rather than disregarding vitamin E as a means to protect against atherosclerosis progression , it would be wiser to better design clinical trials based on current knowledge of the biological properties of the molecule . 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 . 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 . 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 . Cooperation between retinoic acid and phorbol esters enhances human teratocarcinoma differentiation . This study explored cooperation between the retinoic acid ( RA ) and protein kinase C ( PKC ) pathways during differentiation of the multipotential human teratocarcinoma ( TC ) cell line NTERA-2 clone D1 ( abbreviated NT2/D1 ) . We report here that , compared to RA treatment alone , RA combined with the PKC stimulator 12-O-tetradecanoylphorbol-13-acetate ( TPA ) enhanced the regulated expression of the immunophenotypic differentiation markers SSEA-3 , a globo-series carbohydrate , and the ganglio-series carbohydrate antigens GD2 and GD3 . Northern analysis and transient transfection assays revealed that TPA co-treatment augmented the RA-induced expression and activation of the RA nuclear receptor-beta ( P10826 ) , one early marker of RA response in NT2/D1 cells . This finding was extended with transient co-transfection experiments using a P17252 expression vector which revealed that the PKC pathway can augment the activation of P10826 by RA . These experiments establish PKC as a modulator of P10826 expression in NT2/D1 cells . Similarly , experiments showed that RA can modulate activation of the PKC-responsive AP-1 complex , a transcription factor rapidly activated by TPA . Northern analysis and transient transfection assays revealed that , compared to TPA treatment alone , RA and TPA augmented the expression and transcriptional activity of AP-1 in NT2/D1 cells . In contrast , transient transfection assays revealed no cooperative effect between RA and TPA in HeLa cells , indicating that this effect in NT2/D1 cells is cell type-specific . In summary , these studies show that stimulation of the PKC second messenger pathway can modulate tumor differentiation and transcriptional activation of a retinoid receptor associated with RA response . 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 stop transfer sequence of the human UDP-glucuronosyltransferase 1A determines localization to the endoplasmic reticulum by both static retention and retrieval mechanisms . Human UDP-glucuronosyltransferase 1A ( P22309 ) isoforms are endoplasmic reticulum ( ER ) -resident type I membrane proteins responsible for the detoxification of a broad range of toxic phenolic compounds . These proteins contain a C-terminal stop transfer sequence with a transmembrane domain ( TMD ) , which anchors the protein into the membrane , followed by a short cytosolic tail ( CT ) . Here , we investigated the mechanism of ER residency of P22309 mediated by the stop transfer sequence by analysing the subcellular localization and sensitivity to endoglycosidases of chimeric proteins formed by fusion of P22309 stop transfer sequence ( TMD/CT ) with the ectodomain of the plasma membrane P01730 reporter protein . We showed that the stop transfer sequence , when attached to C-terminus of the P01730 ectodomain was able to prevent it from being transported to the cell surface . The protein was retained in the ER indicating that this sequence functions as an ER localization signal . Furthermore , we demonstrated that ER localization conferred by the stop transfer sequence was mediated in part by the KSKTH retrieval signal located on the CT . Interestingly , our data indicated that P22309 TMD alone was sufficient to retain the protein in ER without recycling from Golgi compartment , and brought evidence that organelle localization conferred by P22309 TMD was determined by the length of its hydrophobic core . We conclude that both retrieval mechanism and static retention mediated by the stop transfer sequence contribute to ER residency of P22309 proteins . 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 . Alpha-tocopherol prevents cyclosporin A-mediated activation of phospholipase A2 and inhibition of Na+ , K(+)-adenosine triphosphatase activity in cultured hamster renal tubular cells . At concentrations of 0.5 microM and upward , cyclosporin A ( DB00091 ) caused dose-related inhibition of the growth of a hamster renal tubular cell line ( Q86TB3 ATCC ; Q16663 ) in vitro . Inhibition of cell growth was due to the cytotoxic properties of DB00091 which were associated with enhancement of activity of phospholipase A2 ( P04054 ) according to the increased generation of arachidonic acid and lysophosphatidylcholine ( Q16549 ) . Arachidonate per se , at concentrations of up to 20 microM , did not affect the growth of Q86TB3 cells , while cyclooxygenase and P09917 inhibitors failed to protect the cells against the antiproliferative effects of DB00091 . However , Q16549 caused dose-related inhibition of the growth of Q86TB3 cells . Moreover , coincubation with lysophospholipase or DB00163 ( AT , vitamin E ) , a P04054 inhibitory and lysophospholipid-complexing agent , protected the Q86TB3 cells against both DB00091 and Q16549 . The Na+ , K(+)-ATPase activity of Q86TB3 cells was also inhibited by DB00091 , with the enzyme being protected by inclusion of AT or lysophospholipase . Increased activity of P04054 and inhibition of Na+ , K(+)-ATPase preceded cytotoxicity and cytolysis . Excessive production of lysophospholipids and consequent inhibition of Na+ , K(+)-ATPase in renal tubular cells is a possible mechanism of DB00091 -induced nephrotoxicity . The protective effects of AT suggest that this agent may be clinically useful in preventing the renal side effects of DB00091 . 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 . 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 . 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 . 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 ) 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_1101

MH_train_1101

MH_train_1101

interacts_with DB04817?

multiple_choice

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 . 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 . 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 . 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 . PPARgamma activation abolishes LDL-induced proliferation of human aortic smooth muscle cells via SOD-mediated down-regulation of superoxide . Native LDL would be a mitogenic and chemotactic stimulus of VSMC proliferation and differentiation in the atherosclerotic lesion where endothelial disruption occurred . In previous studies , our group investigated the molecular mechanisms by which LDL induces P10145 production and by which PPARalpha activation abolishes LDL effects in human aortic SMCs ( hAoSMCs ) . Herein is the first report of PPARgamma activation by troglitazone ( TG ) exerting its inhibitory effects on LDL-induced cell proliferation via generation not of H(2)O(2) , but of O2(.-) , and the subsequent activation of Erk1/2 in hAoSMCs . Moreover , in this study TG abolished the LDL-accelerated G(1)-S progression to control levels via down-regulation of active cyclinD1/ P11802 and cyclinE/ P24941 complexes and up-regulation of P38936 (Cip1) expression . TG exerted its anti-proliferative effects through the up-regulation of basal superoxide dismutase ( SOD ) expression . This data suggests that the regulation of O2(.-) is located at the crossroads between LDL signaling and cell proliferation . 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 . Angiotensin metabolism in rat stomach wall : prevalence of angiotensin-(1-7) formation . Our view of renin-angiotensin system ( DB01367 ) has changed over the past two decades : new metabolites and pathways have been described ; also the importance of local renin-angiotensin systems became more clearly understood . However , there is relatively scarce information about formation and action of angiotensin peptides in gastrointestinal tract , especially in the stomach . Here , using LC- P19957 -MS method we assessed the metabolism of Ang I in organ bath of rat stomach wall . Additionally we compared the expression of mRNA of angiotensin converting enzymes ( P12821 , Q9BYF1 ) and neprilysin ( NEP ) in the stomach , aorta and renal artery in rats . Despite , similar levels of expression of P12821 and Q9BYF1 mRNA in stomach wall , aorta and renal artery , the absolute amounts of main Ang I metabolites produced by stomach wall ( in ng/mg of dry tissue ) were much lower than that produced by aorta and renal artery . Also , the pattern of angiotensin I metabolites was different : opposite to aorta and renal artery , incubation of Ang I with stomach wall fragments resulted in predominant formation of Ang-(1-7) and relatively lower production of Ang II . In stomach wall both , perindoprilat and tiorphan decreased production of Ang II , but did not influence generation of Ang-(1-7) . In conclusion , we identified Ang-(1-7) as the main product of Ang I conversion in rat stomach wall . The biological role of prevalence of Ang-(1-7) formation in stomach require further investigation . 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 . 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 . 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 .

MH_train_1102

MH_train_1102

MH_train_1102

interacts_with DB00243?

multiple_choice

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 . 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 . NF-κΒ inhibition is ineffective in blocking cytokine-induced P10145 production but O75791 and P42224 inhibitors are effective . OBJECTIVE : In vitro but not in vivo evidence indicates that blockade of NF-κB is effective in reducing inflammation and production of P10145 . We hypothesized that the failure of in vitro experiments to predict in vivo outcome was due to the use of short time periods of observation and the use of single cytokines to stimulate NF-κB . METHODS : P29320 cells with a NF-κB reporter gene or CaCo-2 cells were stimulated with CM ( IL-1-β ; P01375 -α , and IFN-γ ) or individual cytokines in the presence and absence of NF-κB inhibitors , a P42224 inhibitor , and/or a p38 MAPK inhibitor for periods up to 24 h . NF-κB activation , P10145 production , and nitric oxide production were measured . RESULTS : CM-induced P10145 production in P29320 cells was additive to synergistic . CM enhanced production of P10145 at 24 h but not 4 h was independent of NF-κB . The p38 inhibitor SB203580 and the P42224 inhibitor EGCG blocked CM-induced P10145 production at both early and late time periods . The NF-κB inhibitors PDTC and BAY11-7082 were found to increase CM-stimulated P10145 production in Caco-2 cells at 24 h . CONCLUSIONS : Our data suggest an effective strategy to reduce P10145 production is to block p38 or P42224 rather than NF-κB . 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 . 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 . Progressive alterations of cytokeratin expressions in the process of chronic arsenism . Recent studies of an endemic occurrence of chronic arsenism in a limited area on the southwest coast of Taiwan are focusing on its cytokeratin analysis in hopes of tracing the disease 's biochemical expression . Specimens were obtained from uninvolved skin and arsenical cancers including Bowen 's disease , basal cell carcinoma , and squamous cell carcinoma . In this study , we used two-dimensional polyacrylamide gel electrophoresis to analyse cytokeratin expression . Progressive alterations in cytokeratin expression were found in various skin lesions . These include an expression of P08779 in the uninvolved skin ; P08779 and K6 in Bowen 's disease ; and P08779 , K6 and Q04695 in squamous cell carcinoma and basal cell carcinoma . In addition , we found that the P04264 isoelectric variants shifted to more acidic forms with the complete absence of P04264 in basal cell carcinoma . P08779 expression in uninvolved skin indicates that it is nevertheless in a hyperproliferative status . Q04695 was expressed in squamous cell carcinoma and basal cell carcinoma , but not in Bowen 's disease . The progressive impairment of phosphorylation of P04264 and K2 in the process of chronic arsenism provides us with a suitable model for studying the biological significance of phosphorylation in intermediate filaments during chemical carcinogenesis . 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 . Expression pattern and biochemical characteristics of a major epidermal retinol dehydrogenase . The biological functions of vitamin A in the epidermis are mediated by all-trans retinoic acid , which is biosynthesized from retinol in two oxidative reactions . The first step involves enzymatic conversion of retinol to retinaldehyde . The physiological significance and relative contributions of the various retinol dehydrogenases to the oxidation of retinol in epidermal cells remain unclear . We report the characterization of a retinol dehydrogenase/reductase of the SDR superfamily , hRoDH-E2 , which is abundantly expressed in the epidermis , epidermal appendages and in cultured epidermal keratinocytes . Both in live keratinocytes and in isolated keratinocyte microsomes , where the enzyme normally localizes , hRoDH-E2 functions as a bona fide retinol dehydrogenase . In the prevailing oxidative reaction it recognizes both free- and P09455 -bound retinol , and shows preference toward NADP as a co-substrate . In comparison , hRoDH-E2 retinol dehydrogenase activity in the simple epithelial P29320 293 cells is much lower and in CHO cells is non-existent . hRoDH-E2 transcripts are distributed throughout the epidermal layers but are more abundant in the basal cells . In contrast , the protein is detected predominantly in the basal and the most differentiated living layers . Its synthesis is negatively regulated by retinoic acid . The biochemical properties and the differential expression of hRoDH-E2 in the strata where retinoic acid signaling is critical for epidermal homeostasis support a conclusion that hRoDH-E2 bears the characteristics of the major microsomal retinol dehydrogenase activity in the epidermal keratinocytes in physiological circumstances . 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 . Posttranslational regulation of thioredoxin-interacting protein . Q9H3M7 ( Txnip ) is a metabolic regulator , which modulates insulin sensitivity and likely plays a role in type 2 diabetes . We studied the regulation of Txnip in 3T3- Q9NUQ9 adipocytes . Cells were incubated under different conditions and Txnip was measured by immunoblotting . We confirmed that high glucose markedly increases Txnip expression by promoting transcription . P01308 decreases Txnip protein levels . DB00877 under most conditions decreased Txnip , suggesting that P42345 complex-1 is involved . The acute effects of insulin are mainly posttranscriptional ; insulin ( 100 nM ) accelerates Txnip degradation more than tenfold . This effect is cell type specific . It works in adipocytes , preadipocytes and in Q9BTT4 myotubes but not in HepG2 or in P29320 293 cells or in a pancreatic β-cell line . The ubiquitin/proteasome pathway is involved . Degradation of Txnip occurred within 15 min in the presence of 3 nM insulin and overnight with 0.6 nM insulin . Proteasomal Txnip degradation is not mediated by a cysteine protease or an anti-calpain enzyme . Okadaic acid ( OKA ) , an inhibitor of phosphoprotein phosphatases ( pp ) , markedly reduced Txnip protein and stimulated its further decrease by insulin . The latter occurred after incubation with 1 or 1000 nM OKA , suggesting that insulin enhances the phosphorylation of a pp2A substrate . Incubation with 0.1 μM Wortmannin , a P19957 kinase inhibitor , increased Txnip protein twofold and significantly inhibited its insulin-induced decrease . Thus , while OKA mimics the effect of insulin , Wortmannin opposes it . In summary , insulin stimulates Txnip degradation by a P19957 kinase-dependent mechanism , which activates the ubiquitin/proteasome pathway and likely serves to mitigate insulin resistance . IKKalpha enhances human keratinocyte differentiation and determines the histological variant of epidermal squamous cell carcinomas . Squamous cell carcinomas ( SCCs ) of the skin display different clinical features according to their epithelial differentiation grade and histological variant . Understanding the causes of these divergences might increase the curability of SCCs . Therefore , it is important to study the mechanisms of differentiation in keratinocytes . IKK ( O15111 ) alpha is an important protein for epidermal morphogenesis , although the pathways through which it exerts its function are unknown and controversy exists about its role in cancer development . We show that enhanced IKKalpha expression increases both early and terminal differentiation of human keratinocytes through an P12830 -dependent mechanism . Increased expression of IKKalpha in mouse tumorigenic epidermal cells leads to changes in the differentiation pattern of the resulting SCCs , originating a distinct histological variant that resembles the human acantholytic SCC ( ASCC ) variant . Although human ASCCs have an aggressive clinical course and high risk of metastasis , nothing is known about their etiology . We show that human ASCCs , as observed in the counterpart IKKalpha murine tumors , express high levels of both IKKalpha and P12830 , with absence of keratins P04264 and P13645 , usually co-expressed with IKKalpha and P12830 . The tight correlation between the properties of both murine and human ASCC variants strongly suggests that IKKalpha is responsible for the development of this human SCC variant . 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 . 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 . 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 . Autocrine endocannabinoid signaling through P21554 receptors potentiates OX1 orexin receptor signaling . It has been proposed that OX(1) orexin receptors and CB(1) cannabinoid receptors can form heteromeric complexes , which affect the trafficking of OX(1) receptors and potentiate OX(1) receptor signaling to extracellular signal-regulated kinase ( P29323 ) . We have recently shown that OX(1) receptor activity releases high levels of the endocannabinoid 2-arachidonoyl glycerol ( 2-AG ) , suggesting an alternative route for OX(1)-CB(1) receptor interaction in signaling , for instance , in retrograde synaptic transmission . In the current study , we set out to investigate this possibility utilizing recombinant Chinese hamster ovary P04264 cells . 2-AG released from OX(1) receptor-expressing cells acted as a potent paracrine messenger stimulating P29323 activity in neighboring CB(1) receptor-expressing cells . When OX(1) and CB(1) receptors were expressed in the same cells , OX(1) stimulation-induced P29323 phosphorylation and activity were strongly potentiated . The potentiation but not the OX(1) response as such was fully abolished by specific inhibition of CB(1) receptors or the enzyme responsible for 2-AG generation , diacylglycerol lipase ( DAGL ) . Although the results do not exclude the previously proposed OX(1)-CB(1) heteromerization , they nevertheless unequivocally identify DAGL-dependent 2-AG generation as the pivotal determinant of the OX(1)-CB(1) synergism and thus suggest a functional rather than a molecular interaction of OX(1) and CB(1) receptors . 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 . Modulation of translation-initiation in CHO- P04264 cells by rapamycin-induced heterodimerization of engineered eIF4G fusion proteins . Translation-initiation is a predominant checkpoint in mammalian cells which controls protein synthesis and fine-tunes the flow of information from gene to protein . In eukaryotes , translation-initiation is typically initiated at a 7-methyl-guanylic acid cap posttranscriptionally linked to the 5' end of mRNAs . Alternative cap-independent translation-initiation involves 5' untranslated regions ( UTR ) known as internal ribosome entry sites , which adopt a particular secondary structure . Translation-initiating ribosome assembly at cap or IRES elements is mediated by a multiprotein complex of which the initiation factor 4F ( eIF4F ) consisting of eIF4A ( helicase ) , P06730 ( cap-binding protein ) , and eIF4G is a major constituent . eIF4G is a key target of picornaviral protease 2A , which cleaves this initiation factor into eIF4G(Delta) and (Delta)eIF4G to redirect the cellular translation machinery exclusively to its own IRES-containing transcripts . We have designed a novel translation control system ( TCS ) for conditional as well as adjustable translation of cap- and IRES-dependent transgene mRNAs in mammalian cells . eIF4G(Delta) and (Delta)eIF4G were fused C- and N-terminally to the FK506-binding protein ( FKBP ) and the FKBP-rapamycin-binding domain ( FRB ) of the human FKBP-rapamycin-associated protein ( P42345 ) , respectively . DB00877 -induced heterodimerization of eIF4G(Delta)-FKBP and FRB-(Delta)eIF4G fusion proteins reconstituted a functional chimeric elongation factor 4G in a dose-dependent manner . Rigorous quantitative expression analysis of cap- and IRES-dependent SEAP- ( human placental secreted alkaline phosphatase ) and luc- ( Photinus pyralis luciferase ) encoding reporter constructs confirmed adjustable translation control and revealed increased production of desired proteins in response to dimerization-induced heterologous eIF4G in Chinese hamster ovary ( CHO- P04264 ) cells . Noninvasive delivery of small inhibitory RNA and other reagents to pulmonary alveoli in mice . A technically easy , noninvasive means of delivering molecules to alveoli , which act selectively or specifically in the lung , would be experimentally and therapeutically useful . As proof of principle , we took advantage of the spreading ability of pulmonary surface active material ( InfaSurf ) , mixed it with elastase , glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) small inhibitory RNA ( siRNA ) , or all-trans retinoic acid ( DB00755 ) , and instilled microliter amounts of the mixture into the nose of lightly anesthetized mice . One instillation of elastase caused diffuse alveolar destruction ( emphysema ) demonstrating widespread alveolar delivery . A single nasal instillation of P04406 siRNA , compared with scrambled P04406 siRNA , lowered P04406 protein in lung , heart , and kidney by approximately 50-70 % 1 and 7 days later . To test the possibility of lung-specific delivery of a potentially therapeutic drug , we administered DB00755 and monitored its effect on expression of cellular retinol binding protein ( P09455 ) -1 mRNA , whose translation product is a key molecule in retinoid metabolism . Given intranasally , DB00755 elevated P09455 -1 mRNA 4.3-fold in a lung-specific manner . The same dose and dose schedule of DB00755 given intraperitoneally increased P09455 -1 mRNA only approximately 1.8-fold in lung ; intraperitoneally administered DB00755 elevated expression of P09455 -1 mRNA 1.7-fold or more in brain cortex , cerebellum , and testes , thereby increasing the risk of untoward effects . This simple noninvasive technique allows regulation of specific proteins in the lung and lung-specific delivery of reagents of experimental and potentially therapeutic importance . Listeriolysin O secreted by Listeria monocytogenes induces NF-kappaB signalling by activating the O15111 complex . Listeriolysin O ( LLO ) is a pore-forming cytolysin secreted by the pathogen Listeria monocytogenes and is required for its intracellular survival . We recently demonstrated that in endothelial cells , LLO activates the NF-kappaB signalling pathway . In this work , we studied the LLO-induced molecular cascade of NF-kappaB activation with a cellular model extensively used to analyse the signalling pathway of NF-kappaB activation , i.e. the human embryonic kidney P29320 -293 cell line and its derivatives ( transfectants or mutants ) . When the stably transfected derivative P29320 -293 cells expressing IL-1RI were exposed to LLO , a strong NF-kappaB activation was detected , contrasting with other cell lines ( P29320 -293 wild type , P29320 -293.T and COS ) expressing a very low level of IL-1RI . Although a delayed kinetics of LLO-dependent NF-kappaB activation suggests an autocrine or paracrine IL-1-dependent pathway , we found that LLO-dependent NF-kappaB activation did not require the IL-1 protein synthesis nor the interaction with the IL-1RI specific receptor . Herein , we demonstrated that LLO-dependent NF-kappaB activation requires the activation of the O15111 beta ( IKKbeta ) subunit of IKK complex to phosphorylate and degrade cytoplasmic P25963 , a natural inhibitor of NF-kappaB . The activation induced by LLO does not require the adapters MyD88 and IL-1R-associated kinase ( P51617 ) . We suggested that LLO induces a distinct signalling pathway from that of IL-1 and its receptor . Atrial natriuretic peptide binds to P01160 - Q96GN5 receptors in neuroblastoma cells or is degraded extracellularly at the DB00133 - DB00120 bond . P01160 - Q96GN5 receptors for atrial natriuretic peptide ( P01160 ) showed the following rank order of affinity in intact human neuroblastoma cells NB-OK-1 : human P01160 -(99-126) approximately human P01160 -(102-126) approximately rat P01160 -(99-126) ( P04264 17-32 pM ) > human P01160 -(103-126) > porcine brain natriuretic peptide ( DB04899 ) . Analogues truncated at the C-terminal extremity or devoid of a disulphide bridge , such as rat P01160 -(103-123) , rat C- P01160 -(102-121) , rat P01160 -(111-126) , rat P01160 -(99-109) and rat [desCys105,Cys121] P01160 -(104-126) and chicken P23582 , were not recognized . The occupancy of these high affinity P01160 - Q96GN5 receptors led to marked cyclic GMP accumulation in the presence of 3-isobutyl 1-methylxanthine . An ectoenzymic activity , partly shed in the incubation medium , provoked the stepwise release of DB00120 - DB00125 -[125I] DB00135 , DB00125 -[125I] DB00135 and [125I] DB00135 from rat [125I] P01160 -(99-126) , at an optimal pH of 7.0 . Its inhibition by 1,10-phenanthroline , DB00974 and bacitracin but not by thiorphan suggests the contribution of at least one neutral metalloendopeptidase , distinct from EC 3.4.24.11 , for which P01160 showed high affinity . New K103 β3 allele identified in a context of severe neonatal thrombocytopenia . BACKGROUND : A new β3 allele was identified in a severe case of neonatal alloimmune thrombocytopenia ( < 7 × 10(9) /L ) . STUDY DESIGN AND METHODS : Diagnosis was done by use of monoclonal antibody-specific immobilization of platelet ( Q02083 ) antigen for serologic analyses and polymerase chain reaction ( PCR ) -sequence-specific primers ( SSP ) and PCR-restriction fragment length polymorphism ( RFLP ) for genotyping . Direct sequencing of PCR product was done and mutant αIIbβ3 expressed in P29320 -293 cells . RESULTS : Serologic analysis revealed in the maternal serum an anti-human Q02083 alloantigen ( Q9Y251 ) -1a alloantibody associated to an anti-α2β1 . Anti- Q9Y251 -1a alloimmunization diagnosis was confirmed by genotyping showing maternofetal incompatibility . However , investigation of rare Q9Y251 polymorphisms revealed discrepant Q9Y251 -16b assignation between PCR-RFLP and PCR-SSP . Sequencing revealed a new c.385C > A mutation in the β3 coding sequence resulting in a false assignation of the Q9Y251 -16b allele by PCR-RFLP . This mutation leads to a Q103K substitution in mature β3 . The K103-β3 form of the complex was expressed in P29320 -293 cells but did not react with the maternal serum . CONCLUSION : We have characterized a new rare allele ( frequency < 1 % ) of β3 that yields false Q9Y251 -16b genotyping in PCR-RFLP . This new case of false typing assignation emphasizes the necessity to use two genotyping techniques in diagnosis . This particularly applies for rare Q9Y251 polymorphisms when Q02083 phenotyping can not be used . Induced segregation of human syntenic genes by 5-bromodeozyuridine + near-visible light . Chromosome-breaking agents have been used in two different procedures for promoting segregation of syntenic genes on human chromosome 12 . In method A , a human-Chinese hamster cell hybrid containing the single human chromosome 12 was treated either with 5-bromodeoxyuridine BrdU + near-visible light or with X-rays . In method B , normal human fibroblasts were treated with BrdU + near-visible light followed by their fusion with a Chinese hamster glycine-requiring cell mutant CHO- P04264 /gly-A . Since the human complementing gene for serine hydroxymethyltransferase , an enzyme deficient in gly-A , lies on human chromosome 12 , only those hybrids retaining that chromosome can survive the glycine-free medium . Clones isolated from both procedures were analyzed for the loss or retention of four other syntenic genes on chromosome 12 , P60174 , P04406 , LDH B , and PepB . The results demonstrate that method B is much more effective in generating clones with extensive marker losses . In addition , the segregation pattern and frequency obtained in this study provided information on the linear order of P60174 and P04406 on chromosome 12 . RNAi suppression of Bax and Bak enhances viability in fed-batch cultures of CHO cells . Bcl-2 family proteins play a crucial role in the regulation of the mitochondrial pathway that leads to apoptosis . Members of the Bcl-2 family can be divided into the anti-apoptotic proteins such as Bcl-2 and Bcl-X(L) , and the pro-apoptotic proteins such as Bax and Bak and the BH3-only proteins . In this study , siRNA constructs to silence the Bax and Bak genes in Chinese hamster ovary ( CHO ) cells were generated . Stable CHO cell lines in which the expression of Bax and Bak were significantly knocked down were screened by Western blot analysis and confirmed by RT-PCR . CHO cells with both Bax and Bak knocked down showed a clear resistance against cytotoxic lectins and UV irradiation-induced apoptosis . Compared to original CHO- P04264 cells , these cells also survived longer when cultured under extreme conditions such as complete nutrient depletion or in high-osmolality medium . CHO cells with both Bax and Bak genes knocked down displayed an extended lifespan as well as higher viable cell densities in fed-batch cultures , both in adherent form on microcarrier beads and in suspension . The P01579 productivity by a rCHO P01579 cell line in which both Bak and Bax were knocked down increased by 35 % compared to the control cells . These results indicate that the genetic inactivation of Bax and Bak in recombinant CHO cells can be an effective strategy in delaying the onset of apoptosis in batch and fed-batch cultures . Inhibition of chemokine ( CXC motif ) ligand 12/chemokine ( CXC motif ) receptor 4 axis ( P48061 / P61073 ) -mediated cell migration by targeting mammalian target of rapamycin ( P42345 ) pathway in human gastric carcinoma cells . P48061 / P61073 plays an important role in metastasis of gastric carcinoma . DB00877 has been reported to inhibit migration of gastric cancer cells . However , the role of P42345 pathway in P48061 / P61073 -mediated cell migration and the potential of drugs targeting PI3K/ P42345 pathway remains unelucidated . We found that P48061 activated PI3K/Akt/ P42345 pathway in MKN-45 cells . Stimulating CHO- P04264 cells expressing pEGFP-C1- O43739 -PH fusion protein with P48061 resulted in generation of phosphatidylinositol (3,4,5)-triphosphate , which provided direct evidence of activating PI3K by P48061 . Down-regulation of p110β by siRNA but not p110α blocked phosphorylation of Akt and P23443 induced by P48061 . Consistently , p110β-specific inhibitor blocked the P48061 -activated PI3K/Akt/ P42345 pathway . Moreover , P61073 immunoprecipitated by anti-p110β antibody increased after P48061 stimulation and G(i) protein inhibitor pertussis toxin abrogated P48061 -induced activation of PI3K . Further studies demonstrated that inhibitors targeting the PI3K/ P42345 pathway significantly blocked the chemotactic responses of MKN-45 cells triggered by P48061 , which might be attributed primarily to inhibition of mTORC1 and related to prevention of F-actin reorganization as well as down-regulation of active RhoA , Rac1 , and Cdc42 . Furthermore , rapamycin inhibited the secretion of P48061 and the expression of P61073 , which might form a positive feedback loop to further abolish upstream signaling leading to cell migration . Finally , we found cells expressing high levels of cxcl12 were sensitive to rapamycin in its activity inhibiting migration as well as proliferation . In summary , we found that the P42345 pathway played an important role in P48061 / P61073 -mediated cell migration and proposed that drugs targeting the P42345 pathway may be used for the therapy of metastatic gastric cancer expressing high levels of cxcl12 . The 10q25.3-26.1 G protein-coupled receptor gene Q8NDV2 is epigenetically silenced in human gliomas . Loss of heterozygosity ( LOH ) of the entire chromosome 10 is the most frequent genetic alteration in human glioblastoma ( GBM ) . In addition to P60484 / P60484 on 10q23.3 , clustering of partial deletion break-points on 10q25.3-26.1 points to a second suppressor locus . The proposed target gene Q9UGM3 was not confirmed . By somatic deletion mapping of this region , we identified the complementary DNA encoding the human homologue of rat orphan G protein-coupled receptor Q8NDV2 . Q8NDV2 is highly expressed in fetal and adult brain , but frequently reduced or absent in glioma cells and biopsies , due to de novo methylation of its 5' CpG island . Silencing of Q8NDV2 was reversed with 5-aza-deoxycytidine and the histone deacetylase inhibitor trichostatin A . Furthermore , overexpression of Q8NDV2 in P29320 and in U87 glioma cells increased intracellular DB02527 concentration which is considered to induce astrocytic differentiation . Interestingly , we observed concomitant silencing of Q8NDV2 with O6-methylguanine-DNA methyl transferase ( P16455 ) , a DNA repair gene co-localized on 10q25.3-26.1 ( p=0.0001 ) . We conclude that epigenetic silencing is a common mechanism in malignant gliomas that simultaneously inactivates P16455 and Q8NDV2 . The 10q25.3-26.1 region may contain an important epigenetic pathway in brain tumorigenesis . 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 . 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 . 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 . Chemically synthesized SDF-1alpha analogue , N33A , is a potent chemotactic agent for P61073 / P61073 / P61073 -expressing human leukocytes . Stromal cell-derived factor ( SDF ) 1 is a potent chemoattractant for leukocytes through activation of the receptor P61073 / P61073 / P61073 , which is a fusion co-factor for the entry of T lymphocytotropic human immunodeficiency virus type 1 ( HIV-1 ) . This P61073 -mediated HIV-1 fusion can be inhibited by P48061 . Because of its importance in the study of immunity and AIDS , large scale production of P48061 is desirable . In addition to recombinant technology , chemical synthesis provides means by which biologically active proteins can be produced not only in large quantity but also with a variety of designed modifications . In this study , we investigated the binding and function of an SDF-1alpha analogue , N33A , synthesized by a newly developed native chemical ligation approach . Radioiodinated N33A showed high affinity binding to human monocytes , T lymphocytes , as well as neutrophils , and competed equally well with native recombinant SDF-1alpha for binding sites on leukocytes . N33A also showed equally potent chemoattractant activity as native recombinant SDF-1alpha for human leukocytes . Further study with P61073 / P61073 / P61073 transfected P29320 293 cells showed that N33A binds and induces directional migration of these cells in vitro . These results demonstrate that the chemically synthesized SDF-1alpha analogue , N33A , which can be produced rapidly in large quantity , possesses the same capacity as native SDF-1alpha to activate P61073 -expressing cells and will provide a valuable agent for research on the host immune response and AIDS . Non-susceptibility trends among Enterobacteriaceae from bacteraemias in the UK and Ireland , 2001-06 . BACKGROUND : Enterobacteriaceae are common agents of bacteraemia , with Escherichia coli accounting for 20 % of the cases . Reflecting this importance , members of the family constitute 4 of the 12 collection groups in the British Society for Antimicrobial Chemotherapy ( BSAC ) Bacteraemia Surveillance Programme . METHODS : MICs from the BSAC surveillance programme were reviewed , along with bacteraemia reports received by the Health Protection Agency ( Q9Y251 ) via its CoSurv/LabBase system . Isolates with unusual resistances were subjected to molecular analysis . RESULTS : The BSAC and Q9Y251 systems both revealed dramatically increasing resistance to cephalosporins , ciprofloxacin and gentamicin among E. coli and Klebsiella spp. , with cephalosporin resistance largely contingent on the spread of CTX-M extended-spectrum beta-lactamases ( ESBLs ) ; fluoroquinolone resistance also increased among Proteus mirabilis and ESBL-negative E. coli . Carbapenem resistance remained extremely rare , but two Enterobacter spp. , from the same patient in different years , had KPC carbapenemases , while a few isolates had carbapenem resistance contingent upon combinations of beta-lactamase and impermeability , and ertapenem MICs for AmpC-derepressed Enterobacter spp. rose over time . Three new agents-ceftobiprole , doripenem and tigecycline-were tested . DB04918 was broadly active , except against ESBL producers and Klebsiella oxytoca hyperproducing P04264 enzyme , and was variable against AmpC-derepressed Enterobacter spp. and against Proteus vulgaris . DB06211 was more potent than imipenem . Tigecycline was almost universally active against E. coli , but low-level non-susceptibility ( MIC 2 mg/L ) was frequent among Klebsiella spp . CONCLUSIONS : E. coli and Klebsiella spp. showed dramatic shifts , with sharply rising non-susceptibility to cephalosporins , ciprofloxacin and gentamicin . The rise in cephalosporin resistance reflected dissemination of CTX-M ESBLs . Carbapenems remain broadly active and new agents offer potential .

MH_train_1103

MH_train_1103

MH_train_1103

interacts_with DB01268?

multiple_choice

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 . Telocytes in liver regeneration : possible roles . Telocytes ( TCs ) are a novel type of interstitial cells which are potentially involved in tissue regeneration and repair ( www.telocytes.com ) . Previously , we documented the presence of TCs in liver . However , the possible roles of TCs in liver regeneration remain unknown . In this study , a murine model of partial hepatectomy ( PH ) was used to induce liver regeneration . The number of TCs detected by double labelling immunofluorescence methods ( P28906 / P09619 -α , P28906 / P09619 -ß and P28906 / P08670 ) was significantly increased when a high level of hepatic cell proliferation rate ( almost doubled ) as shown by 5-ethynyl-2'-deoxyuridine ( EdU ) immunostaining and Western Blot of P12004 ( P12004 ) was found at 48 and 72 hrs post-PH . Meanwhile , the number of P08727 positive-hepatic stem cells peaked at 72 hrs post-PH , co-ordinating with the same time-point , when the number of TCs was most significantly increased . Taken together , the results indicate a close relationship between TCs and the cells essentially involved in liver regeneration : hepatocytes and stem cells . It remains to be determined how TCs affect hepatocytes proliferation and/or hepatic stem cell differentiation in liver regeneration . Besides intercellular junctions , we may speculate a paracrine effect via ectovesicles . Exome-level comparison of primary well-differentiated neuroendocrine tumors and their cell lines . Neuroendocrine cancer cell lines are used to investigate therapeutic targets in neuroendocrine tumors ( NET ) and have been instrumental in the design of clinical trials targeting the PI3K/AKT/ P42345 pathways , P15692 inhibitors , and somatostatin analogues . It remains unknown , however , whether the genomic makeup of NET cell lines reflect that of primary NET since comprehensive unbiased genome sequencing has not been performed on the cell lines . Four bronchopulmonary NET ( BP-NET ) -NCI-H720 , NCI-H727 , NCI-H835 , and UMC11-and two pancreatic neuroendocrine tumors ( panNET ) -BON-1 and QGP1-were cultured . DNA was isolated , and exome sequencing was done . GATK and EXCAVATOR were used for bioinformatic analysis . We detected a total of 1,764 nonsynonymous single nucleotide variants at a rate of 8 per Mb in BP-NET and 4.3 per Mb in panNET cell lines , including 52 mutated COSMIC cancer genes in these cell lines , such as P04637 , P38398 , P06400 , P49815 , P46531 , Q09472 , GNAS , P35968 , Q15831 , and P25054 but not P46100 , Q9UER7 , nor O00255 . Our data suggest that mutation rate , the pattern of copy number variations , and the mutational spectra in the BP-NET cell lines are more similar to the changes observed in small cell lung cancer than those found in primary BP-NET . Likewise , mutation rate and pattern including the absence of mutations in P46100 / Q9UER7 , O00255 , and P25490 in the panNET cell lines BON1 and QGP1 suggest that these cell lines do not have the genetic signatures of a primary panNET . These results suggest that results from experiments with BP-NET and panNET cell lines need to be interpreted with caution . P35968 translocates to the nucleus to regulate its own transcription . Vascular Endothelial Growth Factor Receptor-2 ( P35968 ) is the major mediator of the angiogenic effects of P15692 . In addition to its well known role as a membrane receptor that activates multiple signaling pathways , P35968 also has a nuclear localization . However , what P35968 does in the nucleus is still unknown . In the present report we show that , in endothelial cells , nuclear P35968 interacts with several nuclear proteins , including the Sp1 , a transcription factor that has been implicated in the regulation of genes needed for angiogenesis . By in vivo chromatin immunoprecipitation ( ChIP ) assays , we found that P35968 binds to the Sp1-responsive region of the P35968 proximal promoter . These results were confirmed by EMSA assays , using the same region of the P35968 promoter . Importantly , we show that the P35968 DNA binding is directly linked to the transcriptional activation of the P35968 promoter . By reporter assays , we found that the region between -300/-116 relative to the transcription start site is essential to confer P35968 -dependent transcriptional activity . It was previously described that nuclear translocation of the P35968 is dependent on its activation by P15692 . In agreement , we observed that the binding of P35968 to DNA requires P15692 activation , being blocked by DB00112 and DB01268 , two anti-angiogenic agents that inhibit P35968 activation . Our findings demonstrate a new mechanism by which P35968 activates its own promoter that could be involved in amplifying the angiogenic response . [ GIST refractory to imatinib treatment ] . After the introduction of imatinib , the outcome for patients with advanced gastro-intestinal stromal tumor ( GIST ) was vastly improved . However , resistance to imatinib has become a new problem . The cause of resistance to imatinib is the low sensitivity of gene mutations in the P10721 gene or PDGFRα , or an acquisition of additional mutations , and a low plasma level of imatinib . DB01268 is the first drug that showed an effectiveness for treating GIST refractory to imatinib . New molecular target drugs for overcoming imatinib resistance are being developed now . 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 . Second line therapies for the treatment of gastrointestinal stromal tumor . PURPOSE OF REVIEW : Most gastrointestinal stromal tumors eventually acquire resistance to imatinib mesylate . This review focuses on recent progress on management of patients whose disease progresses on the standard dose of imatinib . RECENT FINDINGS : Approximately 30 % of patients failing standard-dose imatinib achieve disease stabilization with high-dose imatinib , but objective responses are few and the clinical benefit usually short-lived . Patients receiving enzyme-inducing drugs may need high imatinib doses to achieve therapeutic blood concentrations . Surgical excision of a single growing metastasis leads to a median progression-free survival time of 7-11 months . DB01268 malate is effective following imatinib failure . The median time to disease progression is approximately 6 months with sunitinib therapy versus 6 weeks with placebo following discontinuation of imatinib , but few ( 5 % ) patients achieve objective response . Patients with gastrointestinal stromal tumor with P10721 exon 9 mutation may benefit more from sunitinib than those with exon 11 mutation . DB01268 frequently causes abnormal thyroid function . SUMMARY : DB01268 is now the approved second line therapy following imatinib failure and for patients intolerant to imatinib . The clinical benefit is only moderate , and thyroid function monitoring is required . Several investigational agents are being evaluated for imatinib-resistant gastrointestinal stromal tumor . Palliative procedures , such as hepatic arterial embolization , also require study . 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 . Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5-linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter ---- MLVI-2 ---- cen ---- P07686 ---- P00374 ---- Pi227- --- cp12.6 ---- ( P05113 , P05112 ) ---- P08700 ---- P04141 ---- P05230 ---- ( P07333 , P09619 ) ---- ( treC,ADRBR ) ---- ( Q5SW96 - Q13585 , P09603 ) ---- qter . The suggested order and orientation for the closely linked P08700 / P04141 gene pair is cen ---- 5' P08700 3' ---- 5' P04141 3' ---- qter , on the basis of analysis of the P04141 rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q- analyses , was telomeric to P04141 and centromeric to P07333 / P09619 , near P05230 . Long-range restriction-enzyme analysis of 5q- DNAs did not detect rearrangements of 5q-linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12.6 , P08700 , and P07333 can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions . 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 . 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 . 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 . 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 . In-silico screening and in-vitro validation of Vascular Endothelial Growth Factor Receptor-2 ( P35968 ) inhibitors . P35968 tyrosine kinase receptor draws attention of the scientific fraternity in drug discovery for its important role in cancer , cardiopulmonary , cardiovascular diseases etc . Hence there is a need for novel P35968 inhibitors screening and testing for their biological activities . The 3D-structure was collected from PDB and stability was checked by using WHATIF and PROCHECK programs and subjected for virtual screening on Zinc database . We used virtual screening method to screen new P35968 blocker molecules based on their binding energies and then docked with active site on the receptor with the help of AUTODOCK software . Based on the results obtained top three molecules ( VRB1-3 ) were selected and tested in Cardiomyocytes H9c2 cells for cell viability under hypoxic condition . The invitro studies showed VRB2 as the best molecule among the selected three molecules as well as with a standard commercial drug DB01268 . 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 . DB01268 treatment in pediatric patients with advanced GIST following failure of imatinib . BACKGROUND : DB01268 inhibits P10721 and other members of the split-kinase-domain family of receptor tyrosine kinases . DB01268 prolongs survival in adult patients with imatinib-resistant gastrointestinal stromal tumor ( GIST ) . We report the experience with sunitinib in pediatric patients with advanced GIST following failure of imatinib . PROCEDURE : DB01268 therapy was provided through a treatment-use protocol . Patients were 10-17 years old at enrollment . All patients had GIST resistant to imatinib therapy . DB01268 was administered daily for 4 weeks in 6-week treatment cycles . P10721 and platelet-derived growth factor receptor alpha ( P16234 ) genotyping of tumor tissue were performed . RESULTS : One patient achieved a partial response , five patients had stable disease and one patient had progressive disease on sunitinib . The duration of disease stabilization was between 7 and 21+ months , with a mean of 15 months . Time to tumor progression was longer on sunitinib than on prior imatinib treatment for five of six patients . Two patients experienced grade 3 adverse events . All other adverse events were grade 1-2 . None of the five patients tested had mutations in P10721 or P16234 . CONCLUSION : DB01268 treatment was associated with substantial initial antitumor activity and acceptable tolerability in this group of pediatric patients with imatinib-resistant GIST . Pharmacogenetics in chemotherapy of colorectal cancer . Although in recent years , chemotherapeutic options for colorectal carcinoma have expanded , overall response rates are still too low , with high rates of toxicity . Pharmacogenetics aim at predicting both treatment response and adverse effects in individual patients . This review describes the current knowledge of pharmacogenetic markers in the systemic treatment of colorectal cancer . P22309 *28 leads to reduced conjugation of SN-38 , the active metabolite of irinotecan , resulting in an increased rate of adverse effects , especially neutropenia . To a lesser extent , increased DB00544 toxicity is predicted by Q12882 *2A . A variable number of tandem repeats polymorphism in the thymidylate synthase enhancer region , in combination with a single nucleotide polymorphism C > G , may predict poorer response to DB00544 . Efficacy of oxaliplatin is influenced by polymorphisms in components of DNA repair systems , such as P07992 and P18887 . Polymorphic changes in the endothelial growth factor receptor probably predict cetuximab efficacy . Furthermore , the antibody-depended cell-mediated cytotoxic effect of cetuximab may be reduced by polymorphisms in the immunoglobin G fragment C receptors . DB00112 efficacy is suspected to be influenced by polymorphisms in the P15692 gene and the hypoxia inducible factor 1alpha gene . Although the interpretation of pharmacogenetic studies is complicated , results imply a promising way of pretreatment prediction of chemotherapy efficacy and toxicity . 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 . Platelets promote osteosarcoma cell growth through activation of the platelet-derived growth factor receptor-Akt signaling axis . The interactions of tumor cells with platelets contribute to the progression of tumor malignancy , and the expression levels of platelet aggregation-inducing factors positively correlate with the metastatic potential of osteosarcoma cells . However , it is unclear how tumor-platelet interaction contributes to the proliferation of osteosarcomas . We report here that osteosarcoma-platelet interactions induce the release of platelet-derived growth factor ( PDGF ) from platelets , which promotes the proliferation of osteosarcomas . Co-culture of platelets with MG63 or Q9UKB1 osteosarcoma cells , which could induce platelet aggregation , enhanced the proliferation of each cell line in vitro . Analysis of phospho-antibody arrays revealed that co-culture of MG63 cells with platelets induced the phosphorylation of platelet derived growth factor receptor ( P09619 ) and Akt . The addition of supernatants of osteosarcoma-platelet reactants also increased the growth of MG63 and Q9UKB1 cells as well as the level of phosphorylated- P09619 and -Akt . DB01268 or LY294002 , but not erlotinib , significantly inhibited the platelet-induced proliferation of osteosarcoma cells , indicating that PDGF released from platelets plays an important role in the proliferation of osteosarcomas by activating the P09619 and then Akt . Our results suggest that inhibitors that specifically target osteosarcoma-platelet interactions may eradicate osteosarcomas . 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 . Molecular-based choice of cancer therapy : realities and expectations . Current choice of cancer therapy is usually empirical and relies mainly on the statistical prediction of the treatment success . Molecular research provides some opportunities to personalize antitumor treatment . For example , life-threatening toxic reactions can be avoided by the identification of subjects , who carry susceptible genotypes of drug-metabolizing genes ( e.g. P51580 , P22309 , P42898 , Q12882 ) . Tumor sensitivity can be predicted by molecular portraying of targets and other molecules associated with drug response . Tailoring of antiestrogen and trastuzumab therapy based on hormone and P04626 receptor status has already become a classical example of customized medicine . Other predictive markers have been identified both for cytotoxic and for targeted therapies , and include , for example , expression of TS , TP , Q12882 , OPRT , P07992 , P16455 , P11388 , class III beta-tubulin molecules as well as genomic alterations of P00533 , P10721 , P00519 oncogenes . [ 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 . [ Strategy for patients with GIST after failure of imatinib ] . DB01268 malate(SU11248)is an oral multitargeted receptor tyrosine kinase inhibitor(MTI)that has antitumor activities for patients with gastrointestinal stromal tumor ; GIST after failure of Imatinib . DB01268 has demonstrated significant clinical benefits , including PFS , RR and OS in the USA and Japan . However , cis-mutations in the activation loop of the P10721 gene tend to develop DB01268 -resistant GIST . Two clinical trials revealed that new multitargeted receptor tyrosine kinase inhibitors , DB00398 and DB04868 , had antitumor activities for DB01268 -resistant GIST with longer PFS and a different spectrum . Now , clinical trials of several new MTIs are ongoing in Western countries . Inhibition of the P10721 gene cis-mutations and antiangiogenesis activities may be essential for the strategy for Imatinib/Sunitinibresistant GIST . Blocking autocrine P15692 signaling by sunitinib , an anti-cancer drug , promotes embryonic stem cell self-renewal and somatic cell reprogramming . Maintaining the self-renewal of embryonic stem cells ( ESCs ) could be achieved by activating the extrinsic signaling , i.e. , the use of leukemia inhibitory factor ( P15018 ) , or blocking the intrinsic differentiation pathways , i.e. , the use of GSK3 and MEK inhibitors ( 2i ) . Here we found that even in medium supplemented with P15018 , mESCs still tend to differentiate toward meso-endoderm lineages after long-term culture and the culture spontaneously secretes vascular endothelial growth factors ( VEGFs ) . Blocking P15692 signaling with sunitinib , an anti-cancer drug and a receptor tyrosine kinase ( RTK ) inhibitor mainly targeting P15692 receptors ( VEGFRs ) , is capable of maintaining the mESCs in the undifferentiated state without the need for feeder cells or P15018 . DB01268 facilitates the derivation of mESCs from blastocysts , and the mESCs maintained in sunitinib-containing medium remain pluripotent and are able to contribute to chimeric mice . DB01268 also promotes iPSC generation from MEFs with only Oct4 . Knocking down P35968 or blocking it with neutralizing antibody mimicks the effect of sunitinib , indicating that blocking P15692 /VEGFR signaling is indeed beneficial to the self-renewal of mESCs . We also found that hypoxia-inducible factor alpha ( HIF1α ) and endoplasmic reticulum ( ER ) stress are involved in the production of P15692 in mESCs . Blocking both pathways inhibits the expression of P15692 and prevents spontaneous differentiation of mESCs . Interestingly , P15018 may also exert its effect by downregulating HIF1α and ER stress pathways and subsequent P15692 expression . These results indicate the existence of an intrinsic differentiation pathway in mESCs by activating the autocrine P15692 signaling . Blocking P15692 signaling with sunitinib or other small molecules help to maintain the mESCs in the ground state of pluripotency . Pleiotropic stromal effects of vascular endothelial growth factor receptor 2 antibody therapy in renal cell carcinoma models . The benefits of inhibiting vascular endothelial growth factor ( P15692 ) signaling in cancer patients are predominantly attributed to effects on tumor endothelial cells . Targeting non-endothelial stromal cells to further impact tumor cell growth and survival is being pursued through the inhibition of additional growth factor pathways important for the survival and/or proliferation of these cells . However , recent data suggest that P15692 receptor ( VEGFR ) -specific inhibitors may target lymphatic vessels and pericytes in addition to blood vessels . Here , in fact , we demonstrate that DC101 ( 40 mg/kg , thrice a week ) , an antibody specific to murine P35968 , significantly reduces all three of these stromal components in subcutaneous ( SKRC-29 ) and orthotopic ( 786-O-LP ) models of renal cell carcinoma ( RCC ) established in nu/nu athymic mice . DB01268 ( 40 mg/kg , once daily ) , a receptor tyrosine kinase inhibitor of P35968 and other growth factor receptors , also caused significant loss of tumor blood vessels in RCC models but had weaker effects than DC101 on pericytes and lymphatic vessels . In combination , sunitinib did not significantly add to the effects of DC101 on tumor blood vessels , lymphatic vessels , or pericytes . Nevertheless , sunitinib increased the effect of DC101 on tumor burden in the SKRC-29 model , perhaps related to its broader specificity . Our data have important implications for combination therapy design , supporting the conclusion that targeting P35968 alone in RCC has the potential to have pleiotropic effects on tumor stroma . Imatinib mesylate induces quiescence in gastrointestinal stromal tumor cells through the CDH1- Q13309 - P46527 signaling axis . Gastrointestinal stromal tumors ( GIST ) are caused by activating mutations in the P10721 or platelet-derived growth factor receptor alpha receptor tyrosine kinase genes . Approximately 85 % of GIST patients treated with imatinib mesylate achieve disease stabilization , however , often in the presence of residual tumor masses . Complete remissions are rare and a substantial proportion of patients develop resistance to imatinib . Our study was designed to determine whether imatinib-associated responses may account for these clinical findings . We report here that imatinib stimulates cellular quiescence in a proportion of GIST cells as evidenced by up-regulation of the CDK inhibitor p27(Kip1) , loss of cyclin A , and reduced BrdUrd incorporation . Mechanistically , these events are associated with an imatinib-induced modulation of the P25054 /CDH1 signaling axis . Specifically , we provide evidence that imatinib down-regulates Q13309 and that this event is associated with increased nuclear CDH1 , an activator of the P25054 that has been shown to regulate Q13309 stability . We also show that those GIST cells that do not undergo apoptosis in response to imatinib overexpress nuclear p27(Kip1) , indicating that they have withdrawn from the cell cycle and are quiescent . Lastly , we provide evidence that a fraction of primary GISTs with high Q13309 expression levels may have an increased risk of disease progression . Taken together , our results support a model in which GIST cells that do not respond to imatinib by apoptosis are removed from the proliferative pool by entering quiescence through modulation of the P25054 /CDH1- Q13309 -p27(Kip1) signaling axis . These results encourage further studies to explore compounds that modulate this pathway as antitumor agents in GISTs . [ Gastrointestinal stromal tumors ] . Gastrointestinal stromal tumors ( GIST ) are the most common sarcomas of the gastrointestinal tract . They affect all segments of the digestive tract . They develop from the interstitial cells of Cajal . Mutations in the Kit gene is present in 86 % of cases and in P09619 gene in 15 % of cases . The marker CD 117 is present in 95 % of cases . Surgery is the standard treatment in localized forms . The tyrosine kinase inhibitor , imatinib is standard in first-line metastatic gastrointestinal stromal tumors , as well as adjuvant treatment after surgery . DB01268 is the standard in second line . 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 . DB01268 deregulates tumor adaptation to hypoxia by inhibiting HIF-1alpha synthesis in HT-29 colon cancer cells . DB01268 ( SU11248 , Sutent ) is a class III/V receptor tyrosine kinase ( RTK ) inhibitor that exhibits potent anti-angiogenic and anticancer activities . Preclinical studies demonstrated that the sunitinib effects are attributed to inhibition of VEGFR and P09619 phosphorylation . However , even in colon cancer cells lacking sunitinib-targeted RTKs , sunitinib effectively inhibits tumor growth in a xenograft model , and this raises a question about the mechanism underlying the in vivo anticancer action of sunitinib . Since hypoxia is a critical microenvironment that tumors face , we addressed the possibility that sunitinib deregulates tumor adaptation to hypoxia . First we found that sunitinib limits the colony growth of HT-29 , which is a colon adenocarcinoma cell line lacking the RTKs , and that HIF-1alpha in the colonies is decreased by sunitinib . In cultured HT-29 cells , sunitinib suppressed HIF-1alpha under hypoxic conditions . Moreover , sunitinib repressed the activity of HIF-1alpha and subsequently decreased the expressions of Q9BYW2 downstream genes . Mechanistically , sunitinib blocked the 5'-UTR-dependent translation of HIF-1alpha . The HIF-1alpha suppression by sunitinib was also reproduced in a P40337 -null renal cell carcinoma cell line , where HIF-1alpha is not degradable . In conclusion , the sunitinib inhibition of Q9BYW2 signaling could restrain tumor progression in hypoxic regions , which may contribute to anticancer effect of sunitinib . Systemic therapy for advanced renal cell carcinoma . Renal cell carcinoma ( RCC ) accounts for approximately 3 % of all cancers and is refractory to cytotoxic chemotherapy - immunotherapy has until recently been the standard of care for advanced disease . Randomised trials reported in the last 5 years have demonstrated that a number of agents including the monoclonal antibody , bevacizumab , and the kinase inhibitors - sorafenib sunitinib , temsirolimus and everolimus - are active in advanced RCC . DB00112 is directed against the vascular endothelial growth factor ( P15692 ) , a key mediator of angiogenesis , whilst sorafenib and sunitinib inhibit a number of targets including the P15692 and platelet-derived growth factor ( P09619 ) receptor tyrosine kinases . DB06287 and everolimus inhibit the intracellular mammalian target of rapamycin ( P42345 ) kinase . DB01268 and temsirolimus have demonstrated efficacy in comparison with immunotherapy in the first-line setting in patients with favourable and poor prognosis advanced disease respectively . In the second-line setting , everolimus has shown benefit over placebo in patients who progress following treatment with a P15692 receptor tyrosine kinase inhibitor and sorafenib has demonstrated efficacy in comparison with placebo in patients with immunotherapy-refractory disease . We review here recent clinical trial data and discuss future developments in the systemic treatment of RCC including combination and sequential therapy , adjuvant therapy , the role of biomarkers and the prospects for the development of rational mechanism-directed therapy in this disease . 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 . Inhibition of activated receptor tyrosine kinases by DB01268 induces growth arrest and sensitizes melanoma cells to DB00188 by blocking Akt pathway . Despite the use of multiple therapeutic strategies , metastatic melanoma remains a challenge for oncologists . Thus , new approaches using combinational treatment may be used to try to improve the prognosis of this disease . In this report , we have analyzed the expression of receptor tyrosine kinases ( RTKs ) in melanoma specimens and in four metastatic melanoma cell lines . Both melanoma specimens and cell lines expressed RTKs , suggesting that they may represent eventual targets for multitargeted tyrosine kinase inhibitor , Suntinib . DB01268 reduced the proliferation of two melanoma cell lines ( M16 and M17 ) and increased apoptosis in one of them ( M16 ) . Moreover , the two metastatic melanoma cell lines harbored an activated receptor ( PDGFRα and VEGFR , respectively ) , and DB01268 suppressed the phosphorylation of the RTKs and their downstream targets Akt and ribosomal protein S6 , in these two cell lines . Similar results were obtained when either PDGFRα or P35968 expression was silenced by lentiviral-mediated short-hairpin RNA delivery in M16 and M17 , respectively . To evaluate the interaction between DB01268 and DB00188 , median dose effect analysis using MTT assay was performed , and combination index was calculated . DB00188 synergistically enhanced the DB01268 -induced growth arrest in DB01268 -sensitive cells ( combination index < 1 ) . Moreover , LY294002 , a PI3K inhibitor , sensitized melanoma cells to DB00188 treatment , suggesting that downregulation of phospho-Akt by DB01268 mediates the synergy obtained by DB00188 + DB01268 cotreatment . Altogether , our results suggest that melanoma cells harboring an activated RTK may be clinically responsive to pharmacologic RTK inhibition by DB01268 , and a strategy combining DB01268 and DB00188 , may provide therapeutic benefit . DB01268 in malignant melanoma : a treatment option only for P10721 -mutated patients ? DB01268 has previously been reported to be potentially effective in the treatment of malignant melanomas expressing c- P10721 . Here we report on the case of a 77-year-old gentleman affected by a metastatic clear cell carcinoma of the kidney and a metastatic malignant melanoma with liver and lung metastases . Despite the negativity for CD117 and the absence of P10721 amplification or mutations in the melanoma specimen , he achieved a partial response both in the lungs and in the liver while on sunitinib ( 50 mg once/day , 4 weeks on/2 weeks off ) for the treatment of kidney cancer . To our knowledge , this represents the first evidence of sunitinib activity in P10721 wild-type melanoma . Further studies should be performed to confirm these preliminary data . DB00171 released from astrocytes during swelling activates chloride channels . DB00171 release from astrocytes contributes to calcium ( [ Ca(2+) ] ) wave propagation and may modulate neuronal excitability . In epithelial cells and hepatocytes , cell swelling causes DB00171 release , which leads to the activation of a volume-sensitive Cl(-) current ( I(Cl,swell) ) through an autocrine pathway involving purinergic receptors . Astrocyte swelling is counterbalanced by a regulatory volume decrease , involving efflux of metabolites and activation of I(Cl,swell) and K(+) currents . We used whole cell patch-clamp recordings in cultured astrocytes to investigate the autocrine role of DB00171 in the activation of I(Cl,swell) by hypo-osmotic solution ( Q9UKB1 ) . Apyrase , an DB00171 /ADP nucleotidase , inhibited Q9UKB1 -activated I(Cl,swell) , whereas DB00171 and the P2Y agonists , ADPbetaS and ADP , induced Cl(-) currents similar to I(Cl,swell) . Neither the P2U agonist , UTP nor the P2X agonist , alpha,beta-methylene DB00171 , were effective . BzATP was less effective than DB00171 , suggesting that Q99572 receptors were not involved . P2 purinergic antagonists , suramin , Q08999 , and pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid ( PPADS ) reversibly inhibited activation of I(Cl,swell) , suggesting that DB00171 -activated P47900 receptors . Thus DB00171 release mediates I(Cl,swell) in astrocytes through the activation of P47900 -like receptors . The multidrug resistance protein ( MRP ) transport inhibitors probenicid , indomethacin , and MK-571 all potently inhibited I(Cl.swell) . DB00171 release from astrocytes in Q9UKB1 was observed directly using luciferin-luciferase and MK-571 reversibly depressed this Q9UKB1 -induced DB00171 efflux . We conclude that DB00171 release via MRP and subsequent autocrine activation of purinergic receptors contributes to the activation of I(Cl,swell) in astrocytes by Q9UKB1 -induced swelling . 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 . 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 . DB01268 as a second-line therapy for advanced GISTs after failure of imatinib : relationship between efficacy and tumor genotype in Korean patients . BACKGROUND : To assess the efficacy and safety of sunitinib with regards to primary genotypes of tumor in Korean patients with advanced gastrointestinal stromal tumors ( GISTs ) who failed an initial therapy of imatinib . METHODS : Clinical data were collected from 88 consecutive patients with metastatic/unresectable GISTs treated with sunitinib at the Asan Medical Center . RESULTS : The median time-to-progression ( TTP ) and overall survival ( OS ) times were 7.1 months and 17.6 months , respectively . Of the 74 patients tested for P10721 ( exons 9 , 11 , 13 , 17 ) and P16234 ( exons 12 and 18 ) , patients with P10721 exon 9 mutant GIST ( n = 11 , 14.9 % ) showed numerically better clinical benefit ( objective response or stable disease ≥ 24 weeks ) rate ( 63.6 % vs 46.8 % , p = 0.504 ) and TTP ( median 13.6 mo vs 6.9 mo , p = 0.631 ) than those with P10721 exon 11 mutant GIST ( n = 47 , 63.5 % ) . The most common grade 3/4 adverse events included neutropenia ( 34.1 % ) , thrombocytopenia ( 33.0 % ) and hand-foot skin reaction ( 25.0 % ) . CONCLUSIONS : DB01268 is an effective and safe second-line therapy for Korean patients with advanced GIST . The superior efficacy of sunitinib against GISTs with P10721 exon 9 mutations appears to be similar in Korean patients to Western experience although statistical significance was not secured . Ligand-dependent platelet-derived growth factor receptor ( P09619 ) -alpha activation sensitizes rare lung cancer and sarcoma cells to P09619 kinase inhibitors . Platelet-derived growth factor ( PDGF ) receptors ( P09619 ) and their ligands play critical roles in several human malignancies . DB01268 is a clinically approved multitargeted tyrosine kinase inhibitor that inhibits vascular endothelial growth factor receptor , c- P10721 , and P09619 , and has shown clinical activity in various solid tumors . Activation of P09619 signaling has been described in gastrointestinal stromal tumors ( P16234 mutations ) as well as in chronic myeloid leukemia ( P11274 - P16234 translocation ) , and sunitinib can yield clinical benefit in both settings . However , the discovery of P09619 activating mutations or gene rearrangements in other tumor types could reveal additional patient populations who might benefit from treatment with anti- P09619 therapies , such as sunitinib . Using a high-throughput cancer cell line screening platform , we found that only 2 of 637 tested human tumor-derived cell lines show significant sensitivity to single-agent sunitinib exposure . These two cell lines [ a non-small-cell lung cancer ( NSCLC ) and a rhabdomyosarcoma ] showed expression of highly phosphorylated P16234 . In the sunitinib-sensitive adenosquamous NSCLC cell line , P16234 expression was associated with focal PFGRA gene amplification , which was similarly detected in a small fraction of squamous cell NSCLC primary tumor specimens . Moreover , in this NSCLC cell line , focal amplification of the gene encoding the P09619 ligand Q9NRA1 was also detected , and silencing P16234 or Q9NRA1 expression by RNA interference inhibited proliferation . A similar codependency on P16234 and Q9NRA1 was observed in the sunitinib-sensitive rhabdomyosarcoma cell line . These findings suggest that , in addition to gastrointestinal stromal tumors , rare tumors that show Q9NRA1 -mediated P16234 activation may also be clinically responsive to pharmacologic P16234 or Q9NRA1 inhibition . Coronary microvascular pericytes are the cellular target of sunitinib malate-induced cardiotoxicity . DB01268 malate is a multitargeted receptor tyrosine kinase inhibitor used in the treatment of human malignancies . A substantial number of sunitinib-treated patients develop cardiac dysfunction , but the mechanism of sunitinib-induced cardiotoxicity is poorly understood . We show that mice treated with sunitinib develop cardiac and coronary microvascular dysfunction and exhibit an impaired cardiac response to stress . The physiological changes caused by treatment with sunitinib are accompanied by a substantial depletion of coronary microvascular pericytes . Pericytes are a cell type that is dependent on intact platelet-derived growth factor receptor ( P09619 ) signaling but whose role in the heart is poorly defined . DB01268 -induced pericyte depletion and coronary microvascular dysfunction are recapitulated by CP-673451 , a structurally distinct P09619 inhibitor , confirming the role of P09619 in pericyte survival . Thalidomide , an anticancer agent that is known to exert beneficial effects on pericyte survival and function , prevents sunitinib-induced pericyte cell death in vitro and prevents sunitinib-induced cardiotoxicity in vivo in a mouse model . Our findings suggest that pericytes are the primary cellular target of sunitinib-induced cardiotoxicity and reveal the pericyte as a cell type of concern in the regulation of coronary microvascular function . Furthermore , our data provide preliminary evidence that thalidomide may prevent cardiotoxicity in sunitinib-treated cancer patients . 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 . Combined environmental risk assessment for 5-fluorouracil and capecitabine in Europe . An environmental risk assessment ( ERA ) was made for the old cytostatic active pharmaceutical ingredient 5-fluorouracil ( DB00544 ) and for capecitabine ( CAP ) , which is a prodrug of DB00544 . This ERA is based on published and company internal data as well as new test results for physicochemical , human metabolism , biodegradability , environmental partitioning and fate , and acute and chronic ecotoxicity properties of the active substance DB00544 as well as on use sales data for DB00544 and CAP in Europe . Predicted environmental concentrations ( PECs ) were extrapolated following the EMEA 2006 Guideline on ERA for human pharmaceuticals and the European Union 2003 Technical Guidance Document ( TGD ) for risk assessment as well as the TGD-based application EUSES v2.0 . Actual amounts sold were taken from IMS Health Databases , in order to refine the default use and EMEA penetration factor as well as the PECs . Moreover , available measured environmental concentrations ( MECs ) were used to supplement PECs . A predicted no-effect concentration ( PNEC ) for DB00544 was derived from chronic ecotoxicity data . Except for the simplistic EMEA Phase I default PEC , the risk characterization by PEC:PNEC and Q9NRJ3 :PNEC ratios for various environmental compartments resulted in no significant risk . As the EMEA Phase I PEC does not integrate documented human metabolism and environmental degradation , in contrast to refined PEC derivations , it is inferred that the current use of CAP and DB00544 does not present any evident risk to the environment . An additional evaluation of persistence , bioaccumulation , and toxicity ( P10721 ) properties supports the conclusion of no significant environmental risk for DB00544 and CAP . 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 . Comparison of dynamic contrast-enhanced MR , ultrasound and optical imaging modalities to evaluate the antiangiogenic effect of PF-03084014 and sunitinib . Noninvasive imaging has been widely applied for monitoring antiangiogenesis therapy in cancer drug discovery . In this report , we used different imaging modalities including high-frequency ultrasound ( HFUS ) , dynamic contrast enhanced-MR ( DCE-MR ) , and fluorescence molecular tomography ( Q96DP5 ) imaging systems to monitor the changes in the tumor vascular properties after treatment with γ-secretase inhibitor PF-03084014 . DB01268 was tested in parallel for comparison . In the MDA-MB-231Luc model , we demonstrated that antiangiogenesis was one of the contributing mechanisms for the therapeutic effect of PF-03084014 . By immunohistochemistry and FITC-lectin perfusion assays , we showed that the vascular defects upon treatment with PF-03084014 were associated with Notch pathway modulation , evidenced by a decrease in the HES1 protein and by the changes in P35968 and HIF1α levels , which indicates down-stream effects . Using a 3D power Doppler scanning method , ultrasound imaging showed that the % vascularity in the MDA-MB-231Luc tumor decreased significantly at 4 and 7 days after the treatment with PF-03084014 . A decrease in the tumor vessel function was also observed through contrast-enhanced ultrasound imaging with microbubble injection . These findings were consistent with the PF-03084014-induced functional vessel changes measured by suppressing the K(trans) values using DCE- Q9BWK5 . In contrast , the Q96DP5 imaging with the AngioSence 680EX failed to detect any treatment-associated tumor vascular changes . DB01268 demonstrated an outcome similar to PF-03084014 in the tested imaging modalities . In summary , ultrasound and DCE-MR imaging successfully provided longitudinal measurement of the phenotypic and functional changes in tumor vasculature after treatment with PF-03084014 and sunitinib . Downsizing treatment with tyrosine kinase inhibitors in patients with advanced gastrointestinal stromal tumors improved resectability . BACKGROUND : Gastrointestinal stromal tumors ( GISTs ) express the receptor tyrosine kinase P10721 . Most GISTs have mutations in the P10721 or P16234 gene , causing activation of tyrosine kinase . Imatinib , a tyrosine kinase inhibitor ( TKI ) , is the first-line palliative treatment for advanced GISTs . DB01268 was introduced for patients with mutations not responsive to imatinib . The aim was to compare the survival of patients with high-risk resected GISTs treated with TKI prior to surgery with historical controls and to determine if organ-preserving surgery was facilitated . METHODS : Ten high-risk GIST-patients had downsizing/adjuvant TKI treatment : nine with imatinib and one with sunitinib . The patients were matched with historical controls ( n = 89 ) treated with surgery alone , from our population-based series ( n = 259 ) . Mutational analysis of P10721 and P16234 was performed in all cases . The progression-free survival was calculated . RESULTS : The primary tumors decreased in mean diameter from 20.4 cm to 10.5 cm on downsizing imatinib . Four patients with R0 resection and a period of adjuvant imatinib had no recurrences versus 67 % in the historical control group . Four patients with residual liver metastases have stable disease on continuous imatinib treatment after surgery . One patient has undergone reoperation with liver resection . The downsizing treatment led to organ-preserving surgery in nine patients and improved preoperative nutritional status in one patient . CONCLUSIONS : Downsizing TKI is recommended for patients with bulky tumors with invasion of adjacent organs . DB01268 can be used for patients in case of imatinib resistance ( e.g. , wild-type GISTs ) , underlining the importance of mutational analysis for optimal surgical planning . [ 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 . 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 . New paradigms in gastrointestinal stromal tumour management . BACKGROUND : Targeted agents have improved the prognosis for patients with advanced gastrointestinal stromal tumours ( GISTs ) . Many patients exhibit intolerance or resistance to first-line therapy with imatinib mesylate . DB01268 malate is approved multinationally for the treatment of advanced imatinib-refractory GIST . DESIGN : This article reviews responses to imatinib and sunitinib reported in clinical trials in advanced GIST and discusses the effect of mutational status on treatment responses ; therapeutic developments in GIST treatment are also reviewed . RESULTS : Imatinib 400 mg/day has shown efficacy for first-line treatment of advanced GIST , particularly in patients with P10721 exon 11 mutations . DB01268 50 mg/day ( Schedule 4/2 ) has demonstrated effectiveness and tolerability in imatinib-refractory GIST , including patients who would be excluded from clinical trials . DB01268 is associated with longer median overall survival in patients with primary P10721 exon 9 mutations and wild-type GIST compared with P10721 exon 11 mutations in a retrospective study . Ongoing studies , including imatinib in the adjuvant setting and the use of targeted agents in sequence or in combination , will further refine the therapeutic pathway for advanced GIST . CONCLUSIONS : The availability of targeted therapies and greater knowledge of the effect of mutational status on patient responses will assist in optimising outcomes in 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 . A phase II study of sunitinib in patients with locally advanced or metastatic cervical carcinoma : NCIC CTG Trial IND.184 . OBJECTIVE : Vascular endothethial growth factor ( P15692 ) and stem cell factor ( c- P10721 ) signaling may play a role in the development and progression of cervical carcinoma . DB01268 malate is an oral , multi-targeted tyrosine kinase inhibitor that inhibits receptors for P15692 , c-Kit and platelet-derived growth factor . This multi-centre phase II study was performed to evaluate the activity of sunitinib in women with locally advanced or metastatic cervical carcinoma who had received up to one prior line of chemotherapy for advanced disease . METHODS : DB01268 , 50 mg/day , was administered in 6-week cycles ( 4 weeks on followed by 2 weeks off treatment ) . The primary endpoint was the objective response rate . RESULTS : Sixteen ( 84 % ) of 19 patients enrolled had stable disease ( median duration 4.4 months , 2.3-17 months ) , but no objective response was observed . Median time to progression was 3.5 months ( range , 2.7-7.0 months ) . Four patients had fistulae develop on study treatment and an additional patient developed a fistula 3.5 months after discontinuation of therapy . All five patients had received either prior chemoradiation or radiation . CONCLUSIONS : A higher rate of fistula formation ( 26.3 % ) was observed than would be expected and is of concern . DB01268 has insufficient activity as a single agent in cervical cancer to warrant further investigation . The behavior of stem cells and progenitor cells in the periodontal ligament during wound healing as observed using immunohistochemical methods . BACKGROUND AND OBJECTIVE : The aim of this study was to identify stem cells or progenitor cells in the periodontal ligament and to investigate their behavior during wound healing of bone defects created experimentally in the alveolar process . MATERIAL AND METHODS : Intradentinal cavities were created in the mesial root of the first molar of 25 adult male rats that were killed 1 , 3 , 5 , 7 and 14 d after surgery . At each time-point , sections were stained immunohistochemically for CD44s ( standard ) , P28906 , c- P10721 , P12004 , Cbfa-1 and 5-bromo-2-deoxyuridine using primary antibodies . For morphometric analysis , the ratios of Cbfa-1 and P12004 -positive cells were calculated from the total number of positive cells/10(4) microm(2) in the cavities . RESULTS : 5-Bromo-2-deoxyuridine-positive cells were observed in the periodontal ligament and had migrated into the wound areas . A small number of CD44s- , P28906 - and c- P10721 -positive cells were observed in the bone marrow , but none were observed in the periodontal ligament . CD44s-positive cells were only observed in the alveolar bone cavity at 5 d after surgery . P28906 - and c- P10721 -positive cells were only observed in the dentin cavity at 7 d after surgery . Cbfa-1 and P12004 scores tended to show an increase 7 d after surgery . CONCLUSION : Mesenchymal stem cells and hematopoietic stem cells in the bone marrow are not involved in the regeneration of the periodontium . Cells that migrated from the residual periodontal ligament regenerated new alveolar bone at the early stage , and the regeneration around the dentin in the cavity was later than in other parts . Peroxisome proliferator-activated receptors ( Q07869 ) agonists affect cell viability , apoptosis and expression of cell cycle related proteins in cell lines of glial brain tumors . The nuclear receptors PPARs ( peroxisome proliferator-activated receptors ) are transcription factors activated by specific ligands . PPARs play an important role in carcinogenesis , inflammation , atherosclerosis , lipid metabolism and diabetes . There is evidence that activation of PPARs by specific ligands is able to suppress the growth of different types of human cancer by mechanisms including the growth arrest , apoptosis and induction of differentiation , although the detailed signalling pathways have not been completely elucidated to date . The aim of our study was to determine whether synthetic ligands of PPARalpha and PPARgamma could affect the viability , proliferation , differentiation , apoptosis and expression of some cell cycle related proteins in glial tumor cell lines . The study was performed on human glioblastoma cell lines U-87 MG , T98G , A172 and U-118 MG . Cell lines were treated by ligands of PPARalpha ( bezafibrate , gemfibrozil ) and PPARgamma ( ciglitazone ) . MTT , flow cytometry , TUNEL assay and immunoblotting were used for detection of changes in cell viability , proliferation , differentiation and apoptosis . DB01393 , ciglitazone and gemfibrozil inhibited viability of glioblastoma cell lines . The synthetic ligands significantly reduced or induced the expression of cyclins , P46527 , p21Waf1/Cip1 , MDM-2 , Bcl-2 , Bax , PARP , Caspase 3 , androgen receptors , etc. and did not affect the expression of the differentiation marker P14136 . Flow cytometry confirmed arrest of the cell cycle although the detection of apoptosis was controversial . Apart from hypolipidemic and hypoglycaemic effects , Q07869 ligands may also have significant cytostatic effects of potential use in anticancer 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 . 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 . DB01268 malate for the treatment of renal cell carcinoma . INTRODUCTION : Over the past decade , a greater understanding into the molecular pathogenesis of renal cell carcinoma ( RCC ) has led to major advances in treatment options . DB01268 is an oral , small-molecule , multi-targeted receptor tyrosine kinase inhibitor ( TKI ) that targets a number of receptors , including vascular endothelial growth factor receptors ( VEGFR ) and platelet-derived growth factor receptors ( P09619 ) . DB01268 was one of the first targeted agents studied in metastatic RCC ( mRCC ) and is currently used worldwide in the management of mRCC . AREAS COVERED : This drug evaluation addresses the preclinical and clinical development of sunitinib . It provides an in-depth discussion of the Phase II data that led to its approval in mRCC and the subsequent Phase III clinical trial comparing sunitinib to interferon-α . More recent data from the large international expanded access trial , in non-clear cell carcinoma patients , different dosing schedule studies and safety issues are also discussed . Finally , areas for the future use of sunitinib , including in the adjuvant setting , are reviewed . EXPERT OPINION : Since the FDA approved sunitinib for advanced RCC in January 2006 , much more has been learned about its efficacy and tolerability . Over the past decade of its clinical use , it has become clear that expertise is required when prescribing sunitinib , in terms of maximizing dose , anticipating and managing side effects , and assessing responses . In the future , a better understanding of sunitinib 's role compared with other P15692 TKIs and P42345 inhibitors , and in other roles such as the adjuvant setting or in non-clear cell pathology , will become evident . DB01268 and periodic hair depigmentation due to temporary c- P10721 inhibition . 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 .

MH_train_1104

MH_train_1104

MH_train_1104

interacts_with DB00898?

multiple_choice

Constitutive NF-kappaB activation confers interleukin 6 ( P05231 ) independence and resistance to dexamethasone and Janus kinase inhibitor DB08877 in murine plasmacytoma cells . Myeloma cells are dependent on P05231 for their survival and proliferation during the early stages of disease , and independence from P05231 is associated with disease progression . The role of the NF-κB pathway in the P05231 -independent growth of myeloma cells has not been studied . Because human herpesvirus 8-encoded P13646 selectively activates the NF-κB pathway , we have used it as a molecular tool to examine the ability of the NF-κB pathway to confer P05231 independence on murine plasmacytomas . We demonstrated that ectopic expression of P13646 , but not its NF-κB-defective mutant or a structural homolog , protected plasmacytomas against P05231 withdrawal-induced apoptosis and resulted in emergence of P05231 -independent clones that could proliferate long-term in vitro in the absence of P05231 and form abdominal plasmacytomas with visceral involvement when injected intraperitoneally into syngeneic mice . These P05231 -independent clones were dependent on NF-κB activity for their survival and proliferation but were resistant to dexamethasone and DB08877 , a selective P23458 /2 inhibitor . Ectopic expression of human T cell leukemia virus 1-encoded Tax protein , which resembles P13646 in inducing constitutive NF-κB activation , similarly protected plasmacytoma cells against P05231 withdrawal-induced apoptosis . Although P13646 is known to up-regulate P05231 gene expression , its protective effect was not due to induction of endogenous P05231 production but instead was associated with sustained expression of several antiapoptotic members of the Bcl2 family upon P05231 withdrawal . Collectively , these results demonstrate that NF-κB activation can not only promote the emergence of P05231 independence during myeloma progression but can also confer resistance to dexamethasone and DB08877 . Genetic variation in three candidate genes and nicotine dependence , withdrawal and smoking cessation in hospitalized patients . AIMS : This study evaluates the relationship of six polymorphisms found in the P32297 , P14416 and P21964 genes with nicotine dependence , the ability to quit smoking and the occurrence of withdrawal symptoms after short-term use of nicotine patch in hospitalized patients . MATERIALS & METHODS : The study included 233 participants from a double-blind , placebo-controlled trial of nicotine patch substitution with a 6-month follow-up period . DB00184 dependence was assessed by the Fagerström Test for DB00184 Dependence ( FTND ) questionnaire , withdrawal symptoms by the Minnesota DB00184 Withdrawal Scale questionnaire and smoking cessation by self-reported abstinence at 1 week , 1 month and 6 months after treatment . RESULTS : After correcting for multiple testing , three polymorphisms in the P14416 gene ( Taq1A , Taq1B and Pro319Pro ) were significantly associated with nicotine dependence ( p = 0.018 , p = 0.048 and p = 0.006 , respectively ) . Using a cutoff point for the FTND score , the P32297 Tyr215Tyr ( rs1051730 ) polymorphism was also associated with nicotine dependence ( p = 0.037 and p = 0.074 after correction for multiple testing ) . No association of any of the studied polymorphisms was observed with either smoking cessation or the occurrence of withdrawal symptoms . CONCLUSION : This study confirms the reported association of the P32297 locus with nicotine dependence and shows the involvement of two independent P14416 polymorphisms in nicotine dependence . 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 . 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 . 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 . 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 . Transcriptome analysis of CNS immediately before and after the detection of P04156 (Sc) in Q04837 /1 sheep scrapie . Sheep scrapie is a transmissible spongiform encephalopathy ( TSE ) , progressive and fatal neurodegenerative diseases of the central nervous system ( CNS ) linked to the accumulation of misfolded prion protein , P04156 (Sc) . New Zealand Cheviot sheep , homozygous for the VRQ genotype of the PRNP gene are most susceptible with an incubation period of 193 days with Q04837 /1 scrapie . However , the earliest time point that P04156 (Sc) can be detected in the CNS is 125 days ( D125 ) . The aim of this study was to quantify changes to the transcriptome of the thalamus and obex ( medulla ) at times immediately before ( D75 ) and after ( D125 ) P04156 (Sc) was detected . Affymetrix gene arrays were used to quantify gene expression in the thalamus and Illumina DGE-tag profiling for obex . Ingenuity Pathway Analysis was used to help describe the biological processes of scrapie pathology . Neurological disease and Cancer were common Bio Functions in each tissue at D75 ; inflammation and cell death were major processes at D125 . Several neurological receptors were significantly increased at D75 ( e.g. Q15825 , Q13255 , Q9UL51 ) , which might be clues to the molecular basis of psychiatric changes associated with TSEs . No genes were significantly differentially expressed at both D75 and D125 and there was no progression of events from earlier to later time points . This implies that there is no simple linear progression of pathological or molecular events . There seems to be a step-change between D75 and D125 , correlating with the detection of P04156 (Sc) , resulting in the involvement of different pathological processes in later TSE disease . P01375 alpha mediates GABA(A) receptor trafficking to the plasma membrane of spinal cord neurons in vivo . The proinflammatory cytokine TNFα contributes to cell death in central nervous system ( CNS ) disorders by altering synaptic neurotransmission . TNFα contributes to excitotoxicity by increasing P42262 -lacking AMPA receptor ( AMPAR ) trafficking to the neuronal plasma membrane . In vitro , increased AMPAR on the neuronal surface after TNFα exposure is associated with a rapid internalization of GABA(A) receptors ( GABA(A)Rs ) , suggesting complex timing and dose dependency of the CNS 's response to TNFα . However , the effect of TNFα on GABA(A)R trafficking in vivo remains unclear . We assessed the effect of TNFα nanoinjection on rapid GABA(A)R changes in rats ( N = 30 ) using subcellular fractionation , quantitative western blotting , and confocal microscopy . GABA(A)R protein levels in membrane fractions of TNFα and vehicle-treated subjects were not significantly different by Western Blot , yet high-resolution quantitative confocal imaging revealed that TNFα induces GABA(A)R trafficking to synapses in a dose-dependent manner by 60 min . TNFα-mediated GABA(A)R trafficking represents a novel target for CNS excitotoxicity . 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 . Imaging intracellular free Ca2+ concentration using yellow cameleons . Green fluorescent protein ( GFP ) -based fluorescent indicators for Ca(2+) offer significant promise for monitoring Ca(2+) in previously unexplored organisms , tissues , and submicroscopic environments because they are genetically encoded , function without cofactors , can be targeted to any intracellular location , and are bright enough for single-cell imaging . These probes use simple GFP variants , circularly permuted GFP ( cpGFP ) , in which the amino and carboxyl portions have been interchanged and reconnected by short spacers between the original termini , or pairs of GFP variants that permit fluorescence resonance energy transfer ( FRET ) . Yellow cameleons ( YCs ) use FRET between cyan- and yellow-emitting variants of Aequorea GFP ( cyan fluorescent protein [ P27918 ] and yellow fluorescent protein [ YFP ] , respectively ) . YCs are composed of a linear combination of P27918 , calmodulin ( P62158 ) , a glycylglycine linker , the P62158 -binding peptide of myosin light-chain kinase ( M13 ) , and YFP . Binding of Ca(2+) to the P62158 moiety of the YC initiates an intramolecular interaction between the P62158 and the M13 domains , causing the chimeric protein to shift from an extended conformation to a more compact one , thereby increasing the efficiency of FRET from P27918 to YFP . This technique is amenable to emission ratioing , which is more quantitative than single-wavelength monitoring . YC3.60 was engineered to enhance its performance as a fluorescent Ca(2+) indicator and here we describe the use of this cameleon to image rapid changes in intracellular free Ca(2+) concentration ( [Ca(2+)]i ) within HeLa cells . FRET imaging is performed using a laser-scanning confocal microscope . Puerarin alleviates noise-induced hearing loss via affecting PKCγ and GABAB receptor expression . Noise-induced hearing loss ( NIHL ) often results from prolonged exposure to high levels of noise . Our previous study revealed that during the development of NIHL , the expression of protein kinase C γ subunit ( PKCγ ) and GABAB receptor ( GABABR ) was changed within the cochlear nuclear complex ( CNC ) , suggesting that these molecules might be the potential targets for the treatment of NIHL . As an extending study , here we focused on puerarin , a major isoflavonoid extracted from Pueraria lobota , which has been used in the treatment of cardiovascular and cerebrovascular diseases , and investigated whether it could protect against NIHL by acting on PKCγ and GABABR . Transgenic Q99259 -GFP knock-in mice were subjected to the NIHL model and their auditory functions were evaluated by the auditory brainstem response thresholds and distortion product oto-acoustic emission signals . Our results showed that 200mg/kg puerarin treatment ameliorated the thresholds of auditory brainstem response of NIHL mice significantly . Triple immunofluorescence staining and electron microscopy results revealed that GFP-positive neurons in the superficial layers of CNC expressed both PKCγ and Q9UBS5 , and Q99259 -positive terminals contacted PKCγ- or Q9UBS5 -positive neurons . Immunoblotting and RT-PCR results showed that NIHL increased the expression of PKCγ but decreased that of Q9UBS5 and O75899 at both protein and mRNA levels in the CNC . Puerarin significantly attenuated the increased expression of PKCγ but elevated the reduced expression of Q9UBS5 and O75899 after noise exposure . Thus , we provided the first evidence that puerarin ameliorated the auditory functions of NIHL mice , and this effect may be due to its ability to regulate the expression of PKCγ and GABABR . Characterization of a Mycobacterium tuberculosis peptide that is recognized by human P01730 + and CD8+ T cells in the context of multiple HLA alleles . The secreted Mycobacterium tuberculosis 10-kDa culture filtrate protein ( P27918 )10 is a potent T cell Ag that is recognized by a high percentage of persons infected with M. tuberculosis . We determined the molecular basis for this widespread recognition by identifying and characterizing a 15-mer peptide , CFP10(71-85) , that elicited P01579 production and CTL activity by both P01730 (+) and CD8(+) T cells from persons expressing multiple MHC class II and class I molecules , respectively . CFP10(71-85) contained at least two epitopes , one of 10 aa ( peptide T1 ) and another of 9 aa ( peptide Q8NHM4 ) . T1 was recognized by P01730 (+) cells in the context of Q8IUH3 *04 , DR5*0101 , and DQB1*03 , and by CD8(+) cells of A2(+) donors . Q8NHM4 elicited responses by P01730 (+) cells in the context of Q8IUH3 *04 and DQB1*03 , and by CD8(+) cells of B35(+) donors . Deleting a single amino acid from the amino or carboxy terminus of either peptide markedly reduced P01579 production , suggesting that they are minimal epitopes for both P01730 (+) and CD8(+) cells . As far as we are aware , these are the shortest microbial peptides that have been found to elicit responses by both T cell subpopulations . The capacity of CFP10(71-85) to stimulate P01579 production and CTL activity by P01730 (+) and CD8(+) cells from persons expressing a spectrum of MHC molecules suggests that this peptide is an excellent candidate for inclusion in a subunit antituberculosis vaccine . 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 . Reactive oxygen species-dependent P01375 converting enzyme activation through stimulation of P41595 and alpha1D autoreceptors in neuronal cells . A major determinant of neuronal homeostasis is the proper integration of cell signaling pathways recruited by a variety of neuronal and non-neuronal factors . By taking advantage of a neuroectodermal cell line ( 1C11 ) endowed with the capacity to differentiate into serotonergic ( 1C115-HT ) or noradrenergic ( 1C11NE ) neurons , we identified serotonin ( 5-hydroxytryptamine , 5-HT ) - and norepinephrine ( NE ) -dependent signaling cascades possibly involved in neuronal functions . First , we establish that P41595 receptors and 1D adrenoceptors are functionally coupled to reactive oxygen species ( ROS ) synthesis through NADPH oxidase activation in 1C115-HT and 1C11NE cells . This observation constitutes the prime evidence that bioaminergic autoreceptors take part in the control of the cellular redox equilibrium in a neuronal context . Second , our data identify P78536 ( P01375 - Converting Enzyme ) , a member of a disintegrin and metalloproteinase ( ADAM ) family , as a downstream target of the P41595 and 1D receptor-NADPH oxidase signaling pathways . Upon P41595 or 1D receptor stimulation , ROS fully govern P01375 - shedding in the surrounding milieu of 1C115-HT or 1C11NE cells . Third , P41595 and 1Dreceptor couplings to the NADPH oxidase- P78536 cascade are strictly restricted to 1C11-derived progenies that have implemented a complete serotonergic or noradrenergic phenotype . Overall , these observations suggest that P41595 and 1D autoreceptors may play a role in the maintenance of neuron- and neurotransmitter-associated functions . Eventually , our study may have implications regarding the origin of oxidative stress as well as up-regulated expression of proinflammatory cytokines in neurodegenerative disorders , which may relate to the deviation of normal signaling pathways . Nicotinic acetylcholine receptors containing the α6 subunit contribute to ethanol activation of ventral tegmental area dopaminergic neurons . DB00184 and alcohol are often co-abused suggesting a common mechanism of action may underlie their reinforcing properties . Both drugs acutely increase activity of ventral tegmental area ( VTA ) dopaminergic ( DAergic ) neurons , a phenomenon associated with reward behavior . Recent evidence indicates that nicotinic acetylcholine receptors ( nAChRs ) , ligand-gated cation channels activated by ACh and nicotine , may contribute to ethanol-mediated activation of VTA DAergic neurons although the nAChR subtype(s) involved has not been fully elucidated . Here we show that expression and activation of nAChRs containing the α6 subunit contribute to ethanol-induced activation of VTA DAergic neurons . In wild-type ( WT ) mouse midbrain sections that contain the VTA , ethanol ( 50 or 100 mM ) significantly increased firing frequency of DAergic neurons . In contrast , ethanol did not significantly increase activity of VTA DAergic neurons in mice that do not express Q15825 , the gene encoding the α6 nAChR subunit ( α6 knock-out ( KO ) mice ) . DB00898 -induced activity in WT slices was also reduced by pre-application of the α6 subtype-selective nAChR antagonist , α-conotoxin MII[E11A] . When co-applied , ethanol potentiated the response to ACh in WT DAergic neurons ; whereas co-application of ACh and ethanol failed to significantly increase activity of DAergic neurons in α6 KO slices . Finally , pre-application of α-conotoxin MII[E11A] in WT slices reduced ethanol potentiation of ACh responses . Together our data indicate that α6-subunit containing nAChRs may contribute to ethanol activation of VTA DAergic neurons . These receptors are predominantly expressed in DAergic neurons and known to be critical for nicotine reinforcement , providing a potential common therapeutic molecular target to reduce nicotine and alcohol co-abuse . 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 . Molecular and biochemical analysis of calmodulin interactions with the calmodulin-binding domain of plant glutamate decarboxylase . We previously provided what to our knowledge is the first evidence that plant glutamate decarboxylase ( Q99259 ) is a calmodulin ( P62158 ) -binding protein . Here , we studied the Q99259 P62158 -binding domain in detail . A synthetic peptide of 26 amino acids corresponding to this domain forms a stable complex with Ca2+/ P62158 with a 1:1 stoichiometry , and amino acid substitutions suggest that tryptophan-485 has an indispensable role in P62158 binding . Chemical cross-linking revealed specific P62158 / Q99259 interactions even in the absence of Ca2+ . However , increasing KCI concentrations or deletion of two carboxy-terminal lysines abolished these interactions but had a mild effect on P62158 / Q99259 interactions in the presence of Ca2+ . We conclude that in the presence of Ca(2+)-hydrophobic interactions involving tryptophan-485 and electrostatic interactions involving the carboxy-terminal lysines mediate P62158 / Q99259 complex formation . By contrast , in the absence of Ca2+ , P62158 / Q99259 interactions are essentially electrostatic and involve the carboxy-terminal lysines . In addition , a tryptophan residue and carboxy-terminal lysines are present in the P62158 -binding domain of an Arabidopsis Q99259 . Finally , we demonstrate that petunia Q99259 activity is stimulated in vitro by Ca2+/ P62158 . Our study provides a molecular basis for Ca(2+)-dependent P62158 / Q99259 interactions and suggests the possible occurrence of Ca(2+)-independent P62158 / Q99259 interactions . 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 . 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 . 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 . SNP mapping and candidate gene sequencing in the class I region of the HLA complex : searching for multiple sclerosis susceptibility genes in Tasmanians . This study is an extension to previously published work that has linked variation in the human leukocyte antigen ( HLA ) class I region with susceptibility to multiple sclerosis ( MS ) in Australians from the Island State of Tasmania . Single nucleotide polymorphism ( SNP ) mapping was performed on an 865-kb candidate region ( D6S1683-D6S265 ) in 166 Tasmanian MS families , and seven candidate genes [ ubiquitin D ( O15205 ) , olfactory receptor 2H3 ( O95918 ) , gamma-aminobutyric acid B receptor 1 ( Q9UBS5 ) , myelin oligodendrocyte glycoprotein ( Q16653 ) , P30511 , HLA complex group 4 ( HCG4 ) and P17693 ] were resequenced . SNPs tagging the extended MS susceptibility haplotype were genotyped in an independent sample of 356 Australian MS trios and SNPs in the Q16653 gene were significantly over-transmitted to MS cases . We identified significant effects on MS susceptibility of HLA-A*2 ( OR : 0.51 ; P = 0.05 ) and A*3 ( OR : 2.85 ; P = 0.005 ) , and two coding polymorphisms in the Q16653 gene ( V145I : P = 0.01 , OR : 2.2 ; V142L : P = 0.04 , OR : 0.45 ) after full conditioning on HLA- Q8IUH3 . We have therefore identified plausible candidates for the causal MS susceptibility allele , and although not conclusive at this stage , our data provide suggestive evidence for multiple class I MS susceptibility genes . Local renin-angiotensin II systems , angiotensin-converting enzyme and its homologue Q9BYF1 : their potential role in the pathogenesis of chronic obstructive pulmonary diseases , pulmonary hypertension and acute respiratory distress syndrome . P00797 -angiotensin II-aldosterone axis has long been known as a regulator of blood pressure and fluid homeostasis . Yet , local renin-angiotensin II systems have been discovered and novel actions of angiotensin II ( AngII ) have emerged among which its ability to act as a immunomodulator and profibrotic molecule . The enzyme responsible for its synthesis , Angiotensin-converting-enzyme ( P12821 ) , is present in high concentrations in lung tissue . In the present paper , we review data from studies of the past decade that implicate AngII and functional polymorphisms of the P12821 gene that increase P12821 activity with increased susceptibility for asthma and chronic obstructive pulmonary disease ( P48444 ) and for pulmonary hypertension . Moreover , drugs that inhibit the synthesis of AngII ( P12821 inhibitors ) or that antagonize its actions on its receptors ( Angiotensin II receptor blockers -ARBs ) have been shown to provide beneficial effects . Another recent discovery reviewed is the presence of a homologue of P12821 , Q9BYF1 , which cleaves a single amino acid from AngII and forms a heptapeptide with vasodilatory actions , Ang 1-7 . The balance between P12821 and Q9BYF1 is crucial for controlling AngII levels . P12821 and Q9BYF1 also appear to modify the severity of Acute Respiratory Distress Syndrome ( ARDS ) , with Q9BYF1 playing a protective role . Finally , mention is made to the recent discovery of Q9BYF1 as a receptor for the P49591 Corona Virus . P23560 activation of P62158 -kinase kinase via transient receptor potential canonical channels induces the translation and synaptic incorporation of P42261 -containing calcium-permeable AMPA receptors . Glutamatergic synapses in early postnatal development transiently express calcium-permeable AMPA receptors ( CP-AMPARs ) . Although these P42262 -lacking receptors are essential and are elevated in response to brain-derived neurotrophic factor ( P23560 ) , little is known regarding molecular mechanisms that govern their expression and synaptic insertion . Here we show that P23560 -induced P42261 translation in rat primary hippocampal neurons requires the activation of mammalian target of rapamycin ( P42345 ) via calcium calmodulin-dependent protein kinase kinase ( CaMKK ) . Specifically , P23560 -mediated phosphorylation of threonine 308 ( T308 ) in AKT , a known substrate of CaMKK and an upstream activator of P42345 -dependent translation , was prevented by ( 1 ) pharmacological inhibition of CaMKK with STO-609 , ( 2 ) overexpression of a dominant-negative CaMKK , or ( 3 ) short hairpin-mediated knockdown of CaMKK . P42261 surface expression induced by P23560 , as assessed by immunocytochemistry using an extracellular N-terminal P42261 antibody or by surface biotinylation , was impaired following knockdown of CaMKK or treatment with STO-609 . Activation of CaMKK by P23560 requires transient receptor potential canonical ( TRPC ) channels as SKF-96365 , but not the DB01221 receptor antagonist d-APV , prevented P23560 -induced P42261 surface expression as well as phosphorylation of CaMKI , AKT(T308) , and P42345 . Using siRNA we confirmed the involvement of Q9UL62 and Q9Y210 subunits in P23560 -induced AKT(T308) phosphorylation . The P23560 -induced increase in mEPSC was blocked by IEM-1460 , a selected antagonist of CP-AMPARs , as well as by the specific repression of acute P42261 translation via siRNA to P42261 but not P42262 . Together these data support the conclusion that newly synthesized P42261 subunits , induced by P23560 , are readily incorporated into synapses where they enhance the expression of CP-AMPARs and synaptic strength . 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 . Anatomical evidence for glutamatergic transmission in primary sensory neurons and onto postganglionic neurons controlling penile erection in rats : an ultrastructural study with neuronal tracing and immunocytochemistry . In male rats , the dorsal penile nerve ( DPN ) conveys sensory information from the genitals to the lumbosacral spinal segments of the spinal cord . DPN is the afferent limb of a reflex loop that supports reflexive erections , and that includes a network of spinal interneurons and autonomic and somatic motoneurons to the penis and perineal striated muscles . Autonomic efferent pathways to the penis relay in the major pelvic ganglion ( P29372 ) . Glutamate ( DB00142 ) is a likely candidate as a neurotransmitter of reflexive erections . Both AMPA and DB01221 glutamatergic receptor subunits are present in the lumbosacral spinal cord , and AMPA and DB01221 receptor antagonists block reflexive erections . In the present study , we used tract-tracing experiments combined with immunohistochemical and immunocytochemical techniques to ascertain the presence of DB00142 at two different levels of the network controlling reflexive erections . DPN afferents were localized in the dorsal horn of the lumbosacral cord and displayed the characteristics of either C-fibers or Adelta fibers . DPN terminals ( some of them glutamatergic ) were mainly distributed in the medial edge of the dorsal horn in the Q9BTT4 spinal segment . GluR1 subunits were present in some DPN afferents , suggesting that they could be autoreceptors . DPN fibers were also present in the P29372 , as were DB00142 terminals and P48058 subunits . The results reveal the presence of DB00142 in DPN fibers and terminals and suggest that both the spinal cord and the P29372 use glutamatergic transmission to control reflexive erections . 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 . 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 . 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 . Antitumor activity of Triolimus : a novel multidrug-loaded micelle containing Paclitaxel , DB00877 , and 17- P29372 . Triolimus is a first-in-class , multidrug-loaded micelle containing paclitaxel , rapamycin , and 17- P29372 . In this study , we examine the antitumor mechanisms of action , efficacy , and toxicity of Triolimus in vitro and in vivo . In vitro cytotoxicity testing of Triolimus was conducted using two aggressive adenocarcinomas including the lung cancer cell line , A549 , and breast cancer cell line , MDA-MB-231 . The three-drug combination of paclitaxel , rapamycin , and 17- P29372 displayed potent cytotoxic synergy in both A549 and MDA-MB-231 cell lines . Mechanistically , the drug combination inhibited both the Ras/Raf/mitogen-activated protein kinase and PI3K/Akt/ P42345 pathways . Triolimus was advanced into tumor xenograft models for assessment of efficacy , toxicity , and mechanisms of action . In vivo , a three-infusion schedule of Triolimus inhibited A549 and MDA-MB-231 tumor growth far more potently than paclitaxel-containing micelles and effected tumor cures in MDA-MB-231 tumor-bearing animals . Tumor growth delays resulted from a doubling in tumor cell apoptosis and a 50 % reduction in tumor cell proliferation compared with paclitaxel-containing micelles . Enhanced antitumor efficacy was achieved without clinically significant increases in acute toxicity . Thus , Triolimus displays potent synergistic activity in vitro and antitumor activity in vivo with comparable toxicity to paclitaxel . These observations provide strong support for further development of Triolimus and an important proof of concept for safe , effective nanoparticle-based delivery of three complementary anticancer agents . Role of P30532 -A3 genetic Locus variants and developing drug for chronic obstructive pulmonary disease . Cigarette smoking is one of the major risk factors for P48444 and P48444 severity . In turn P48444 is a major independent risk factor for lung cancer . Genome-wide association ( GWA ) studies both in lung cancer and P48444 highlighted the same variants ( SNPs ) on chromosome 15q25 marking the gene cluster P32297 - P30926 - P30532 for these smoking related diseases , showing a stimulating connection between this common genetic region and smoking behavior and smoking related illnesses . Different authors identified two candidate regions associated with age at smoking initiation in patients with P48444 . The nicotinic acetylcholine receptor polymorphism ( rs1051730 ) on chromosome 15q25 is associated with major tobacco-related diseases in the general population with additional increased risk of P48444 as well as lung cancer . Moreover variants on the gene cluster P32297 - P30926 - P30532 are associated with nicotine addiction antismoking therapy and antismoking therapy side-effects . These findings not only support the notion that variants can influence any therapy for smoking cessation , but offer rational bases to develop new drugs and new therapeutic strategies . Scope of Proposed Topic ( 50 words ) : Genome-wide association ( GWA ) studies both in lung cancer and P48444 highlighted the same variants ( SNPs ) on the gene cluster P32297 - P30926 - P30532 . These data not only support the notion that variants can influence any therapy for smoking cessation , but offer rational bases to develop new drugs and new therapeutic strategies . Transient , P41595 receptor-mediated facilitation in neuropathic pain : Up-regulation of PKCγ and engagement of the DB01221 receptor in dorsal horn neurons . Spinal nociception can be facilitated by 5-HT2 receptors in neuropathic pain . We investigated the involvement of glutamate receptors in dorsal neuron hyperexcitation that is promoted by P41595 receptor ( 5-HT2BR ) after spinal nerve ligation ( Q16658 ) in the rat . Augmentation of C-fiber-evoked potentials by spinal superfusion with 5-HT2BR agonist BW 723C86 in nerve-ligated rats was impeded by co-administration of DB01221 receptor ( NMDAR ) antagonist D-AP5 , but not by Q13255 /5 antagonist Q96BJ3 or Q14416 /3 antagonist LY 341495 . Evoked potentials were increased by cis-ACPD in nerve-injured rats , irrespective of simultaneous 5-HT2BR blockade by SB204741 . In uninjured rats , NMDAR agonist cis-ACPD enhanced evoked potentials in the presence of BW 723C86 but not if administered alone or during exposure to protein kinase C γ ( PKCγ ) inhibitor peptide . Triple immunofluorescence labelings revealed co-localization of NMDAR and 5-HT2BR in PKCγ-expressing perikarya in lamina II neurons . As a result of Q16658 , PKCγ was transiently and bilaterally up-regulated in synaptic fraction from dorsal horn homogenates , peaking at day 2 and returning to basal levels by day 9 . Chronic blockade of 5-HT2BR with selective antagonist SB 204741 after Q16658 bilaterally decreased the following : ( i ) PKCγ up-regulation in synaptic fraction , ( ii ) phosphorylation of NMDAR subunit Q9UHB4 ( serine 889 ) in synaptic fraction , and ( iii ) co-localization of both PKCγ and phosphorylated Q9UHB4 with postsynaptic marker P78352 . Chronic delivery of SB 204741 bilaterally attenuated thermal and mechanical allodynia occurring after Q16658 , particularly at day 2 post injury . These findings suggest that transient activation of the PKCγ/NMDAR pathway is critically involved in 5-HT2BR-mediated facilitation in the Q16658 model of neuropathic pain . 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 . 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 . 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 . 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 . 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 . 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 . P00747 activator inhibitor-1 supports P10145 -mediated neutrophil transendothelial migration by inhibition of the constitutive shedding of endothelial P10145 /heparan sulfate/syndecan-1 complexes . The endothelium is the primary barrier to leukocyte recruitment at sites of inflammation . Neutrophil recruitment is directed by transendothelial gradients of P10145 that , in vivo , are bound to the endothelial cell surface . We have investigated the identity and function of the binding site(s) in an in vitro model of neutrophil transendothelial migration . In endothelial culture supernatants , P10145 was detected in a trimolecular complex with heparan sulfate and syndecan-1 . Constitutive shedding of P10145 in this form was increased in the presence of a neutralizing Ab to plasminogen activator inhibitor-1 ( P05121 ) , indicating a role for endothelial plasminogen activator in the shedding of P10145 . Increased shedding of P10145 /heparan sulfate/syndecan-1 complexes was accompanied by inhibition of neutrophil transendothelial migration , and aprotinin , a potent plasmin inhibitor , reversed this inhibition . Platelets , added as an exogenous source of P05121 , had no effect on shedding of the complexes or neutrophil migration . Our results indicate that P10145 is immobilized on the endothelial cell surface through binding to syndecan-1 ectodomains , and that plasmin , generated by endothelial plasminogen activator , induces the shedding of this form of P10145 . P05121 appears to stabilize the chemoattractant form of P10145 at the cell surface and may represent a therapeutic target for novel anti-inflammatory strategies . P32297 variant for lung cancer is associated with chronic obstructive pulmonary disease in Korea . BACKGROUND : Genome-wide association studies have identified P32297 as a lung cancer and chronic obstructive pulmonary disease ( P48444 ) candidate gene in non-Hispanic Caucasian cohorts . However , there are differences in minor allele frequencies among ethnic groups , and limited data exists for Asian populations . OBJECTIVES : The aim of this case-control study was to determine whether there is an association between P48444 and genetic variation in P32297 in the Korean population . In addition , we investigated the association of P32297 with intermediate disease phenotypes including emphysema and lung function in P48444 subjects . METHODS : Two single-nucleotide polymorphisms ( SNPs ) in P32297 ( rs660652 and rs12910984 ) were genotyped in 219 P48444 subjects registered in the Korean Obstructive Lung Disease cohort study and in 305 control subjects . Volumetric computed tomography was performed in all P48444 subjects . Emphysema severity was measured quantitatively by determining the volume fraction of the lung below -950 Hounsfield units . Logistic regression analysis for case-control analysis and linear regression modeling for quantitative analysis were performed using SAS . RESULTS : This case-control analysis of 219 P48444 patients and 305 control participants identified a significant association between an SNP of P32297 ( rs12910984 ) and P48444 ( p = 0.049 ) . Analysis in P48444 subjects revealed that genetic variations were not associated with FEV1 . There was no association between SNPs and emphysema severity . However , both SNPs were significantly associated with DLCO . CONCLUSION : Genetic variations in P32297 are associated with P48444 in the Korean population . Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking . Binge drinking of alcoholic beverages among adolescents is a common practice that can have serious health consequences . DB00898 is a potent immunomodulator that alters a wide range of immune responses . However , it is unclear whether there is a differential immune response to alcoholic beverages with a high versus low concentration of ethanol . In this study , we used a PCR array containing 46 primer pairs of selected genes to compare mRNA expression in the spleen , an immune system organ , of adolescent rats following binge drinking of alcohol solutions containing either 20 % or 52 % ethanol ( v/v , 4.8 g/kg daily dosage ) , or water ( control ) for 3 d . We found that , expression of IL-1β , P05231 , P13500 , and GABA(A) receptor α2 subunit in the spleen were decreased , and P41594 and P46098 receptor expression were increased after administration of an ethanol solution containing 52 % ethanol , but not one with 20 % ethanol . Our data suggest that alcohol-mediated immunomodulatory effects are , in part , dependent on the ethanol by volume concentration . This is the first study to show that exposure to a high ethanol percentage beverage can have more profound effects on immune responses than one with a low percentage of ethanol . 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 . 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 . 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 ) . Low ethanol concentration alters P30532 RNA levels during early human development . DB00898 use is common and consumption during pregnancy has been shown to lead to a myriad of physical and neurologic abnormalities commonly referred to as fetal alcohol spectrum disorder . Substance addiction , which includes alcohol , has been shown to involve the major nicotinic acetylcholine receptor subunit P30532 . Using human embryonic stem cells as a model of early human development , we show that low concentrations of ethanol ( 20mM ) can alter the expression of P30532 . Changes in P30532 expression is linked to altered GABA and DB01221 receptor expression , as well as abnormal development of the frontal cortex . These results suggest that alcohol exposure can alter early neurologic development , which may favor addiction and other developmental abnormalities in unborn children . 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 . 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 . Vaporized perfluorocarbon confers protection against acute lung injury by inhibiting P14780 expression without protective effects in other organs . OBJECTIVE : Vaporized perfluorocarbon ( P27918 ) is a treatment for lung injury ; this study investigated its mode of action and potential protective effects on other organs , which are unclear . METHODS : Acute lung injury was induced by lung lavage with artificial seawater in 32 female New Zealand White rabbits . Animals received either conventional mechanical ventilation ( CMV ) , positive end-expiratory pressure under CMV ( PEEP ) , vaporized P27918 ventilation , or positive end-expiratory pressure with vaporized P27918 ventilation ( PEEP+ P27918 ) . Histopathology of the lung , small intestine , liver and kidney were investigated . Matrix metalloproteinase ( MMP ) -9 mRNA levels in the lung were analysed . RESULTS : Pathological injury of the lung was significantly alleviated in the PEEP , P27918 and PEEP+ P27918 groups compared with the CMV group . Tissue damage in the liver , kidney and small intestine was similar between all groups . P14780 mRNA levels in the PEEP , P27918 and PEEP+ P27918 groups were significantly lower than those in the CMV group . CONCLUSIONS : Vaporized P27918 ventilation can significantly alleviate lung injury but has no significant protective effect on other organs . Alleviation of lung injury may be associated with P14780 inhibition . 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 . 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 . Haploinsufficiency of Cyfip1 produces fragile X-like phenotypes in mice . BACKGROUND : Copy number variation ( CNV ) at the 15q11.2 region , which includes a gene that codes for Q7L576 ( cytoplasmic Q06787 interacting protein 1 ) , has been implicated in autism , intellectual disability and additional neuropsychiatric phenotypes . In the current study we studied the function of Cyfip1 in synaptic physiology and behavior , using mice with a disruption of the Cyfip1 gene . METHODOLOGY/PRINCIPAL FINDINGS : We observed that in Cyfip1 heterozygous mice metabotropic glutamate receptor ( mGluR ) -dependent long-term depression ( LTD ) induced by paired-pulse low frequency stimulation ( PP-LFS ) was significantly increased in comparison to wildtype mice . In addition , mGluR-LTD was not affected in the presence of protein synthesis inhibitor in the Cyfip1 heterozygous mice , while the same treatment inhibited LTD in wildtype littermate controls . mGluR-agonist ( RS ) -3,5-dihydroxyphenylglycine ( DHPG ) -induced LTD was also significantly increased in hippocampal slices from Cyfip1 heterozygous mice and again showed independence from protein synthesis only in the heterozygous animals . Furthermore , we observed that the mammalian Target of DB00877 ( P42345 ) inhibitor rapamycin was only effective at reducing mGluR-LTD in wildtype animals . Behaviorally , Cyfip1 heterozygous mice showed enhanced extinction of inhibitory avoidance . Application of both P41594 and Q13255 antagonist to slices from Cyfip1 heterozygous mice reversed the increase in DHPG-induced LTD in these mice . CONCLUSIONS/SIGNIFICANCE : These results demonstrate that haploinsufficiency of Cyfip1 mimics key aspects of the phenotype of Fmr1 knockout mice and are consistent with the hypothesis that these effects are mediated by interaction of Cyfip1 and Fmrp in regulating activity-dependent translation . The data provide support for a model where Q7L576 haploinsufficiency in patients results in intermediate phenotypes increasing risk for neuropsychiatric 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 . 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 . 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 . Chromosome 8p as a potential hub for developmental neuropsychiatric disorders : implications for schizophrenia , autism and cancer . Defects in genetic and developmental processes are thought to contribute susceptibility to autism and schizophrenia . Presumably , owing to etiological complexity identifying susceptibility genes and abnormalities in the development has been difficult . However , the importance of genes within chromosomal 8p region for neuropsychiatric disorders and cancer is well established . There are 484 annotated genes located on 8p ; many are most likely oncogenes and tumor-suppressor genes . Molecular genetics and developmental studies have identified 21 genes in this region ( ADRA1A , O15013 , Q15822 , Q15825 , Q05901 , Q9UBT3 , Q16555 , Q06889 , O60258 , Q9NP95 , P11362 , FZD3 , LDL , NAT2 , P07197 , Q02297 , PCM1 , P00750 , P48454 , Q8N474 and P54219 / P54219 ) that are most likely to contribute to neuropsychiatric disorders ( schizophrenia , autism , bipolar disorder and depression ) , neurodegenerative disorders ( Parkinson 's and Alzheimer 's disease ) and cancer . Furthermore , at least seven nonprotein-coding RNAs ( microRNAs ) are located at 8p . Structural variants on 8p , such as copy number variants , microdeletions or microduplications , might also contribute to autism , schizophrenia and other human diseases including cancer . In this review , we consider the current state of evidence from cytogenetic , linkage , association , gene expression and endophenotyping studies for the role of these 8p genes in neuropsychiatric disease . We also describe how a mutation in an 8p gene ( Fgf17 ) results in a mouse with deficits in specific components of social behavior and a reduction in its dorsomedial prefrontal cortex . We finish by discussing the biological connections of 8p with respect to neuropsychiatric disorders and cancer , despite the shortcomings of this evidence . 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 . 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 . Association study of 45 candidate genes in nicotine dependence in Han Chinese . Numerous genetic linkages , association studies have been performed in different ethnic groups and revealed many susceptibility loci and genes for nicotine dependence . However , limited similar researches were performed in Han Chinese . This study was designed to investigate the association of candidate genes with nicotine dependence in Han Chinese . We genotyped 384 SNPs within 45 candidate genes with nicotine dependence in a Han Chinese population consisting 223 high nicotine dependent subjects and 257 low nicotine dependent subjects by employing GoldenGate genotyping assay ( Illumina ) . Following association analysis was performed using PLINK software . Individual SNP-based association analysis revealed that nine SNPs located in P35462 ( rs2630351 ) , P21918 ( rs1967550 ) , Q9Y6R4 ( rs2314378 ) , DDC ( rs11575461 ) , Q05901 ( rs4954 ) , O75899 ( rs2779562 ) , P14416 ( rs11214613 and rs6589377 ) and P43681 ( rs2236196 ) were significantly associated with FTND after correction for multiple testing with the p values from 2.59×10(-7) to 9.99×10(-5) . Haplotype-based association analysis revealed haplotype G-A-A formed by rs2630351 , rs167771 and rs324032 and haplotype G-G-G-A formed by rs3773678 , rs2630349 , rs2630351 and rs167771 in P35462 ; haplotype of G-A formed by rs2779562 and rs2808566 in O75899 and haplotype of T-T-A-G-A formed by rs6832644 , rs4057797 , rs9764 , rs4552421 and rs10033119 in P25929 are associated with FTND ( p=3.61×10(-7)-8.78×10(-6) ) . Our results provided confirmation of the previous findings that P14416 , P35462 , DDC , Q05901 , O75899 and P43681 are associated with nicotine dependence . Furthermore , we for the first time report a significant association between nicotine dependence and P21918 , Q9Y6R4 and P25929 . These findings need independent replication in the future studies . 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 . Cortical parvalbumin GABAergic deficits with α7 nicotinic acetylcholine receptor deletion : implications for schizophrenia . Dysfunction of cortical parvalbumin ( PV ) -containing GABAergic interneurons has been implicated in cognitive deficits of schizophrenia . In humans microdeletion of the P36544 ( α7 nicotinic acetylcholine receptor , nAChR ) gene is associated with cortical dysfunction in a broad spectrum of neurodevelopmental and neuropsychiatric disorders including schizophrenia while in mice similar deletion causes analogous abnormalities including impaired attention , working-memory and learning . However , the pathophysiological roles of α7 nAChRs in cortical PV GABAergic development remain largely uncharacterized . In both in vivo and in vitro models , we identify here that deletion of the α7 nAChR gene in mice impairs cortical PV GABAergic development and recapitulates many of the characteristic neurochemical deficits in PV-positive GABAergic interneurons found in schizophrenia . α7 nAChR null mice had decreased cortical levels of GABAergic markers including PV , glutamic acid decarboxylase 65/67 ( Q05329 /67 ) and the α1 subunit of GABAA receptors , particularly reductions of PV and Q99259 levels in cortical PV-positive interneurons during late postnatal life and adulthood . Cortical GABAergic synaptic deficits were identified in the prefrontal cortex of α7 nAChR null mice and α7 nAChR null cortical cultures . Similar disruptions in development of PV-positive GABAergic interneurons and perisomatic synapses were found in cortical cultures lacking α7 nAChRs . Moreover , DB01221 receptor expression was reduced in GABAergic interneurons , implicating DB01221 receptor hypofunction in GABAergic deficits in α7 nAChR null mice . Our findings thus demonstrate impaired cortical PV GABAergic development and multiple characteristic neurochemical deficits reminiscent of schizophrenia in cortical PV-positive interneurons in α7 nAChR gene deletion models . This implicates crucial roles of α7 nAChRs in cortical PV GABAergic development and dysfunction in schizophrenia and other neuropsychiatric disorders . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Matrix metalloproteinase 2 releases active soluble ectodomain of fibroblast growth factor receptor 1 . Recent studies have demonstrated the existence of a soluble fibroblast growth factor ( FGF ) receptor type 1 ( P11362 ) extracellular domain in the circulation and in vascular basement membranes . However , the process of P11362 ectodomain release from the plasma membrane is not known . Here we report that the 72-kDa gelatinase A ( matrix metalloproteinase type 2 , P08253 ) can hydrolyze the Val368-Met369 peptide bond of the P11362 ectodomain , eight amino acids upstream of the transmembrane domain , thus releasing the entire extracellular domain . Similar results were obtained regardless of whether FGF was first bound to the receptor or not . The action of P08253 abolished binding of FGF to an immobilized recombinant P11362 ectodomain fusion protein and to Chinese hamster ovary cells overexpressing P11362 The released recombinant P11362 ectodomain was able to bind FGF after P08253 cleavage , suggesting that the cleaved soluble receptor maintained its FGF binding capacity . The activity of P08253 could not be reproduced by the 92-kDa gelatinase B ( P14780 ) and was inhibited by tissue inhibitor of metalloproteinase type 2 . These studies demonstrate that P11362 may be a specific target for P08253 on the cell surface , yielding a soluble FGF receptor that may modulate the mitogenic and angiogenic activities of FGF . GABA Receptors Genes Polymorphisms and DB00898 Dependence : No Evidence of an Association in an Italian Male Population . OBJECTIVE : The genes encoding for gamma-aminobutyric acid ( GABA ) A and B receptors may be considered as candidates for alcoholism ; genetic alterations at this level may produce structural and functional diversity and thus play a role in the response to alcohol addiction treatment . To investigate these aspects further , we conducted a preliminary genetic association study on a population of Italian male alcohol addicts , focusing on GABA A and B receptors . METHODS : A total of 186 alcohol-dependent subjects ( in the first phase 139 , then 47 more samples ) and 182 controls were genotyped for 25 single nucleotide polymorphisms ( SNPs ) of genes encoding the alpha-1 subunit of GABA A receptor ( P14867 ) and subunits 1 and 2 of GABA B receptor ( Q9UBS5 and O75899 ) . The chi-squared test for allele and genotype distributions and Hardy-Weinberg equilibrium analysis of both subjects and controls were performed . Bonferroni 's correction for multiple comparisons was applied . RESULTS : Preliminary results comparing 139 alcohol-dependent subjects and 182 controls showed differences in genotype distribution in the former for SNP rs29253 , located in the intron region of the Q9UBS5 gene . In order to clarify the meaning of this association , 47 more samples from alcohol-dependent subjects were tested for this SNP only : the previously found association was not confirmed . CONCLUSION : The lack of significant differences between the two groups does not provide evidence that GABRA 1 and Q9UBS5 and 2 genes are candidates for alcoholism in this population . Further studies with larger samples are needed , together with investigation of other components of the GABA pathway .

MH_train_1105

MH_train_1105

MH_train_1105

interacts_with DB00159?

multiple_choice

DB00644 1 directly affects corpora lutea lifespan in Mediterranean buffalo ( Bubalus bubalis ) during diestrus : presence and in vitro effects on enzymatic and hormonal activities . The expression of gonadotropin-releasing hormone ( P01148 ) receptor ( P30968 ) and the direct role of P01148 on corpora lutea function were studied in Mediterranean buffalo during diestrus . Immunohistochemistry evidenced at early , mid , and late luteal stages the presence of P30968 only in large luteal cells and P01148 in both small and large luteal cells . Real-time PCR revealed P30968 and P01148 mRNA at the three luteal stages , with lowest values in late corpora lutea . In vitro corpora lutea progesterone production was greater in mid stages and lesser in late luteal phases , whereas prostaglandin F2 alpha ( PGF2alpha ) increased from early to late stages , and DB00917 was greater in the earlier-luteal phase . Cyclooxygenase 1 ( prostaglandin-endoperoxide synthase 1 ; P23219 ) activity did not change during diestrus , whereas P35354 increased from early to late stages , and DB00917 -9-ketoreductase ( DB00917 -9-K ) was greater in late corpora lutea . P23219 activity was greater than P35354 in early corpora lutea and lesser in late luteal phase . In corpora lutea cultured in vitro , the P01148 analog ( buserelin ) reduced progesterone secretion and increased PGF2alpha secretion as well as P35354 and DB00917 -9-K activities at mid and late stages . DB00917 release and P23219 activity were increased by buserelin only in late corpora lutea . These results suggest that P01148 is expressed in all luteal cells of buffalo , whereas P30968 is only expressed in large luteal phase . Additionally , P01148 directly down-regulates corpora lutea progesterone release , with the concomitant increases of PGF2alpha production and P35354 and DB00917 -9-K enzymatic activities . 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 . 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 . Cardioprotective mechanism of telmisartan via P37231 - P29474 pathway in dahl salt-sensitive hypertensive rats . BACKGROUND : Recently , some investigators have shown that telmisartan , an angiotensin II ( Ang II ) -receptor blocker ( ARB ) , is a partial agonist of the peroxisome proliferator-activated receptor-gamma ( P37231 ) . We investigate whether telmisartan improves cardiovascular remodeling associated with the production of endothelial nitric oxide synthase ( P29474 ) through P37231 , inhibits the Rho-kinase pathway , and suppresses oxidative stress in Dahl salt-sensitive ( DS ) hypertensive rats . METHODS : Telmisartan ( 1 mg/kg per day ) or telmisartan plus P37231 inhibitor , GW9662 ( 1 mg/kg per day ) was administered from the age of 6-11 weeks . Age-matched male Dahl salt-resistant ( DR ) rats served as a control group . RESULTS : The levels of P29474 and P37231 expression , and P29474 phosphorylation were significantly lower in DS rats than in DR rats . Chronic telmisartan treatment in DS rats significantly increased these parameters , but not telmisartan plus GW9662 . Telmisartan effectively inhibited the vascular lesion formation such as medial thickness and perivascular fibrosis , but not telmisartan plus GW9662 . Moreover , upregulated RhoA protein , Rho-kinase mRNA , and myosin light-chain phosphorylation in DS rats was decreased by telmisartan to a similar degree as observed after treatment with Y-27632 , a selective Rho-kinase inhibitor . In addition , NAD(P)H oxidase P13498 , p47phox , gp91phox expression , and mitogen-activated protein kinase and its downstream effector P08133 S6 kinase phosphorylation in DS rats was also inhibited by telmisartan . CONCLUSIONS : These results suggest that the cardioprotective mechanism of telmisartan may be partly due to improvement of endothelial function associated with P37231 - P29474 , oxidative stress , and Rho-kinase pathway . 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 . 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 . 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 . 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 . 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 . 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 . Transforming growth factor-beta ( TGF-beta ) activates cytosolic phospholipase A2alpha ( cPLA2alpha ) -mediated prostaglandin E2 (PGE)2/EP1 and peroxisome proliferator-activated receptor-gamma ( P37231 ) /Smad signaling pathways in human liver cancer cells . A novel mechanism for subversion of TGF-beta-induced mitoinhibition . Transforming growth factor-beta ( TGF-beta ) potently inhibits the growth of human epithelial cells . However , neoplastic epithelial cells become resistant to TGF-beta-mediated mitoinhibition , and the mechanisms for this alteration during tumorigenesis are not fully understood . This study was designed to determine whether there is an association between the cytosolic phospholipase A2alpha ( cPLA2alpha ) -controlled eicosanoid metabolism and the growth response to TGF-beta in human liver cancer cells . TGF-beta treatment induced simultaneous Smad-mediated gene transcription and phosphorylation of cPLA2alpha . Whereas Smad activation inhibited tumor cell growth , phosphorylation of P47712 alpha promoted growth and counteracted Smad-mediated mitoinhibition . TGF-beta1 failed to prevent the growth of cells with high basal expression of cPLA2alpha , but inhibition of P47712 alpha , cyclooxygenase-2 ( P35354 ) , or EP1 receptor restored mitoinhibition by TGF-beta1 in these cells . These results suggest that resistance of tumor cells to TGF-beta-mediated mitoinhibition involves activation of cPLA2alpha/ P35354 /EP1 signaling . Furthermore , the TGF-beta1-induced Smad transcriptional activity and mitoinhibition were blocked by overexpression of cPLA2alpha or peroxisome proliferator-activated receptor-gamma ( P37231 ) but enhanced by depletion of cPLA2alpha or P37231 . These findings , along with the observations that cPLA2alpha activates P37231 and that P37231 binds P84022 , illustrate novel cPLA2alpha/ P35354 /EP1 and cPLA2alpha/ P37231 /Smad signaling pathways that counteract the mitoinhibition by TGF-beta in human cancer cells . 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 . 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 . 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 . P37231 : a nuclear receptor with affinity for cannabinoids . An increasing number of cannabinoid actions are being reported that do not appear to be mediated by either P21554 or CB2 , the known cannabinoid receptors . One such example is the synthetic analog ajulemic acid ( AJA ) , which shows potent analgesic and anti-inflammatory effects in rodents and humans . AJA binds weakly to P21554 only at concentrations many fold higher than its therapeutic range , and is , therefore , completely free of psychotropic effects in both normal subjects and pain patients suggesting the involvement of a target site other than P21554 . AJA as well as several other cannabinoids appear to have profound effects on cellular lipid metabolism as evidenced by their ability to transform fibroblasts into adipocytes where the accumulation of lipid droplets can be readily observed . Such transformations can be mediated by the activation of the nuclear receptor P37231 . A variety of small molecule ligands including AJA have been shown to induce the activation of P37231 and , in some cases this has led to the introduction of clinically useful agents . It is suggested that P37231 may serve a receptor function for certain actions of some cannabinoids . [ 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 . 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 . DB00163 prevents ethanol-induced inflammatory , hormonal , and cytotoxic changes in reproductive tissues . DB00898 causes decreased function of the hypothalamic-pituitary-gonadal ( HPG ) axis . DB00898 resulted in inflammatory changes in HPG manifested by increased concentrations of pro-inflammatory cytokines . Since , such cytokines have deleterious effects on functions of HPG , it seemed possible that ethanol 's suppressive action could be due , at least in part , to this inflammation . Since oxidative stress can cause inflammation , we have used the antioxidant vitamin E to test , whether reducing inflammation might protect reproductive functions from ethanol . Rats were fed an ethanol diet or pair fed identically without ethanol for a 3-week period . For the last 10 days , animals were given 30 IU/kg or 90 IU/kg or vehicle . DB00898 significantly increased hypothalamic , pituitary and testicular P01375 and P05231 , all changes prevented by the higher dose of vitamin E. Also , ethanol induced changes in P01148 , LH , testosterone , and testicular germ cell apoptosis were similarly prevented by vitamin E . These data strikingly show that vitamin E protects the HPG from deleterious effects of ethanol and suggests that the mechanism of this protection might be both anti-inflammatory and antioxidant . Polymorphism Pro12Ala of P37231 in prepubertal children with premature adrenarche and its association with growth in healthy children . BACKGROUND : The peroxisome proliferator-activated receptor-γ2 ( PPARγ2 ) participates in the regulation of insulin sensitivity , and has connections to the GH-IGF system and DB01285 -adrenal androgen axis . We hypothesized that P37231 Pro12Ala polymorphism leading to decreased receptor activity could contribute to the premature onset of adrenarche ( PA ) . METHODS : We performed a cross-sectional association study in 73 prepubertal children with PA and 97 age- and sex-matched healthy control children . Growth data , baseline hormone levels , and the values of oral glucose tolerance test ( OGTT ) were compared with P37231 genotypes . RESULTS : We found no difference in the genotype distribution of P37231 between the PA and control children . The minor Ala12 variant was associated with lower current height SD scores ( SDS ) in the controls , but no similar association was seen in the PA children . Birth measures and current weight for height were equal between the genotype groups in both control and PA children . The Ala12 variant was associated with trends for a lower serum P05019 level and lower serum insulin concentration at the 120-min time point of OGTT in control children . CONCLUSIONS : The Pro12Ala genotype of P37231 was not associated with PA . The minor Ala12 variant was associated with lower height SDS in healthy prepubertal children indicating its possible effects on growth . 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 . 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 . 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 . 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 .

MH_train_1106

MH_train_1106

MH_train_1106

interacts_with DB09036?

multiple_choice

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 . 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 . 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 . 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 . 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 ) 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 . 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 . 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 . DB01095 inhibits raft dependent Fcgamma receptor signalling in human monocytes . Statins inhibit P04035 and thus block cholesterol and isoprenoid biosynthesis . Since statins also have anti-inflammatory effects , we investigated the effect of fluvastatin on monocyte Fcgamma receptor function . DB01095 ( 0.5-20 microM ) inhibited Fcgamma receptor signal transduction at the level of tyrosine kinase activation , in a time and dose dependent manner . Initiation of tyrosine phosphorylation is not thought to involve prenylated proteins ; thus , we hypothesised that fluvastatin might disrupt cholesterol and sphingolipid membrane rafts to impair signalling . Consistent with this hypothesis , fluvastatin decreased ( and mevalonate rescued ) signalling molecules within membrane rafts in parallel with effects on tyrosine phosphorylation events . Raft integrity was unaffected by prenyl transferase inhibitors . In addition , Fcgamma receptor mediated immune complex trafficking , activation of Q96HU1 kinases ( P29323 and p38 ) , and downstream inflammatory mediator release ( P03956 and P05231 ) were blocked by fluvastatin . Thus , P04035 inhibition alters immune receptor signalling by disrupting membrane rafts essential for the initiation of signal transduction . Inhibition of Fcgamma receptor function may limit development and progression of atherosclerosis by decreasing monocyte/macrophage inflammatory mediator release . Since many receptors utilise cholesterol rich rafts this mechanism may have broader significance given the pleiotropic effects of statins . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 ) . 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 . 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 . Neem leaf glycoprotein suppresses regulatory T cell mediated suppression of monocyte/macrophage functions . We have shown that neem leaf glycoprotein ( NLGP ) inhibits the regulatory T cell ( Tregs ) induced suppression of tumoricidal functions of P08571 (+) P34810 (+) monocyte/macrophages ( MO/Mφ ) from human peripheral blood . Cytotoxic efficacy of MO/Mφ toward macrophage sensitive cells , U937 , is decreased in presence of Tregs ( induced ) , however , it was increased further by supplementation of NLGP in culture . Associated Treg mediated inhibition of perforin/granzyme B expression and nitric oxide release from MO/Mφ was normalized by NLGP . Altered status of signature cytokines , like , IL-12 , P22301 , P05231 , TNFα from MO/Mφ under influence of Tregs is also rectified by NLGP . Tregs significantly enhanced the expression of altered marker , mannose receptor ( CD206 ) on P34810 (+) cells that was downregulated upon NLGP exposure . In addition to tumoricidal functions , antigen presenting ability of MO/Mφ is hampered by Treg induced downregulation of P33681 , P42081 and HLA- DB01048 . NLGP upregulated these molecules in MO/Mφ even in the presence of Tregs . Treg mediated inhibition of MO/Mφ chemotaxis in contact dependent manner was also normalized partially by NLGP , where participation of P51681 was documented . Overall results suggest that Treg influenced pro-tumor MO/Mφ functions are rectified in a significant extent by NLGP to create an anti-tumor immune environment . 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 . P04035 inhibitors ( statin ) prevents retinal neovascularization in a model of oxygen-induced retinopathy . PURPOSE : Retinal neovascularization ( RNV ) is a primary cause of blindness and involves the dysfunction of retinal capillaries . Recent studies have emphasized the beneficial effects of inhibitors of P04035 ( statins ) in preventing vascular dysfunction . In the present study , the authors characterized the therapeutic effects of statins on RNV . METHODS : Statin treatment ( 10 mg/kg/d fluvastatin ) was tested in a mouse model of oxygen-induced retinopathy . Morphometric analysis was conducted to determine the extent of capillary growth . Pimonidazole hydrochloride was used to assess retinal ischemia . Western blot and immunohistochemical analyses were used to assess protein expression levels and immunolocalization . Lipid peroxidation and superoxide radical formation were determined to assess oxidative changes . RESULTS : DB01095 treatment significantly reduced the area of the capillary-free zone ( P < 0.01 ) , decreased the formation of neovascular tufts ( P < 0.01 ) , and ameliorated retinal ischemia . These morphologic and functional changes were associated with statin effects in preventing the upregulation of P15692 , Q9BYW2 alpha , phosphorylated P40763 , and vascular expression of the inflammatory mediator P05362 ( P < 0.01 ) . Superoxide production and lipid peroxidation in the ischemic retina were also reduced by statin treatment ( P < 0.01 ) . CONCLUSIONS : These data suggest the beneficial effects of statin treatment in preventing retinal neovascularization . These beneficial effects appear to result from the anti-oxidant and anti-inflammatory properties of statins . 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 . 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 . 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 . 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 . 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 . 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 .

MH_train_1107

MH_train_1107

MH_train_1107

interacts_with DB00030?

multiple_choice

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 . 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 . 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 . 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 . P01308 signaling in mouse oocytes . Continuous exposure of follicles/oocytes to elevated levels of insulin compromises embryonic developmental competence , although the underlying cellular mechanisms are unknown . The objectives of the present study were to determine whether mouse oocytes have insulin receptors and a functional insulin signaling cascade , and whether insulin exposure during oocyte growth or maturation influences meiotic progression and chromatin remodeling . Immunoblot and immunocytochemical analyses of germinal vesicle-intact ( GVI ) oocytes demonstrated the presence of insulin receptor-beta . P06213 expression in oocytes was increased by gonadotropin stimulation , and remained elevated throughout meiotic maturation . Fully grown GVI oocytes contained 3-phosphoinositide-dependent protein kinase-1 ( O15530 ) , thymoma viral proto-oncogene 1 ( P31749 ) , and glycogen synthase kinase 3 ( GSK3 ) . In vitro maturation of GVI oocytes in 5 microg/ml insulin had no influence on meiotic progression or the incidence of normal metaphase II ( MII ) chromosome condensation . Treatment of oocytes during maturation had no effect on P49840 /B protein expression or phosphorylation of S21/9 . However , the culturing of preantral follicles for 10 days with 5 microg/ml insulin increased the phosphorylation of oocyte P49841 , indicating GSK3 inactivation . The rates of development to metaphase I ( MI ) were similar for oocytes obtained from insulin-treated follicles and controls , whereas the incidence of abnormal MI chromatin condensation was significantly higher in oocytes obtained from follicles cultured with insulin compared to those cultured without insulin . These results demonstrate that oocytes contain a functional insulin signaling pathway , and that insulin exposure during oocyte growth results in chromatin remodeling aberrations . These findings begin to elucidate the mechanisms by which chronic elevated insulin influences oocyte meiosis , chromatin remodeling , and embryonic developmental competence . 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 . Igf-I influences everolimus activity in medullary thyroid carcinoma . CONTEXT : Medullary thyroid carcinoma ( P04629 ) is a rare tumor originating from thyroid parafollicular C cells . It has been previously demonstrated that insulin-like growth factor I ( P05019 ) protects P04629 from the effects of antiproliferative drugs . DB01590 , an P42345 inhibitor , has shown potent antiproliferative effects in a human P04629 cell line , TT , and in two human P04629 primary cultures . OBJECTIVE : To verify whether P05019 may influence the effects of everolimus in a group of human P04629 primary cultures . DESIGN : We collected 18 MTCs that were dispersed in primary cultures , treated without or with 10 nM-1 μM everolimus and/or 50 nM P05019 . Cell viability was evaluated after 48 h , and calcitonin ( CT ) secretion was assessed after a 6 h incubation . P08069 downstream signaling protein expression profile was also investigated . RESULTS : DB01590 significantly reduced cell viability in eight P04629 [ by ~20 % ; P < 0.01 vs. control ; everolimus-responders ( E-R ) MTCs ] , while cell viability did not change in 10 MTCs [ everolimus-non-responders ( E-NR ) MTCs ] . In E-R MTCs , P05019 blocked the antiproliferative effects of everolimus that did not affect CT secretion , but blocked the stimulatory effects of P05019 on this parameter . P08069 downstream signaling proteins were expressed at higher levels in E-NR P04629 as compared to E-R MTCs . CONCLUSION : P05019 protects a subset of P04629 primary cultures from the antiproliferative effects of everolimus and stimulates CT secretion by an P42345 mediated pathway that , in turn , may represent a therapeutic target in the treatment of aggressive MTCs . 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 . 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 . A Single 60 mg Dose of DB06643 Might Improve Hepatic P01308 Sensitivity in Postmenopausal Nondiabetic Severe Osteoporotic Women . Background. The O14788 / Q9Y6Q6 / O00300 signaling pathway is crucial for the regulation of osteoclast activity and bone resorption being activated in osteoporosis . The pathway has been also suggested to influence glucose metabolism as observed in chronic low inflammation . Aim . To test whether systemic blockage of O14788 by the monoclonal antibody denosumab influences glucose metabolism in osteoporotic women . Study Design . This is a prospective study on the effect of a subcutaneously injected single 60 mg dose of denosumab in 14 postmenopausal severe osteoporotic nondiabetic women evaluated at baseline and 4 and 12 weeks after their first injection by an oral glucose tolerance test . Results . A single 60 mg dose of denosumab efficiently inhibited serum alkaline phosphatase while it did not exert any significant variation in fasting glucose , insulin , or HOMA-IR at both 4 and 12 weeks . No changes could be detected in glucose response to the glucose load , Matsuda Index , or insulinogenic index . Nonetheless , 60 mg denosumab induced a significant reduction in the hepatic insulin resistance index at 4 weeks and in HbA1c levels at 12 weeks . Conclusions . A single 60 mg dose of denosumab might positively affect hepatic insulin sensitivity though it does not induce clinical evident glucose metabolic disruption in nondiabetic patients . In vitro and in vivo evaluation of insulin-producing beta TC6- P08709 cells in microcapsules . In the present study , the insulin secretory capacity of beta TC6- P08709 cells in microcapsules was evaluated . The cell mass within capsules was found to expand in a three-dimensional fashion , in contrast to cells seeded on plates that grew as a monolayer . In in vitro studies , both free and encapsulated cells were found to secrete insulin in the absence of glucose , at 13.6 +/- 1.1 and 14.5 +/- 0.9 ng.10(6) cells-1.60 min-1 , respectively , with the response rising to a maximum of 26.0 +/- 0.8 and 31 +/- 2.3 ng.10(6) cells-1.60 min-1 in the presence of 16.8 mM glucose . Encapsulated cells were able to produce Ca2+ responses in the presence of DB00761 ( 50 mM ) and BAY K 8644 ( 100 microM ) . In in vivo studies , intraperitoneal transplantation of 3.0 x 10(6) microencapsulated cells into mice ( n = 5 ) with streptozotocin-induced diabetes resulted in the restoration of normoglycemia up to 57 days . P01308 concentrations rose from 0.4 +/- 0.1 ng/ml before the graft administration to 2.2 +/- 0.8 ng/ml after the transplantation in the normoglycemic recipients . An oral glucose challenge in transplant recipients demonstrated a flat glucose response , suggesting extremely high glucose clearance rates . These data demonstrate the potential use of the immunoisolated beta-cell lines for the treatment of diabetes . 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 . 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 . 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 . Diminished phosphodiesterase-8B potentiates biphasic insulin response to glucose . DB02527 activates multiple signal pathways , crucial for the pancreatic beta-cells function and survival and is a major potentiator of insulin release . A family of phosphodiesterases ( PDEs ) terminate the DB02527 signals . We examined the expression of PDEs in rat beta-cells and their role in the regulation of insulin response . Using RT-PCR and Western blot analyses , we identified Q14432 , Q13370 , Q07343 , Q08499 , and O95263 in rat islets and in P01308 -1E cells and several possible splice variants of these PDEs . Specific depletion of Q14432 with small interfering ( si ) RNA ( siPDE3A ) led to a small ( 67 % ) increase in the insulin response to glucose in P01308 -1E cells but not rat islets . siPDE3A had no effect on the glucagon-like peptide-1 ( 10 nmol/liter ) potentiated insulin response in rat islets . Depletion in O95263 levels in rat islets using similar technology ( siPDE8B ) increased insulin response to glucose by 70 % , the potentiation being of similar magnitude during the first and second phase insulin release . The siPDE8B-potentiated insulin response was further increased by 23 % when glucagon-like peptide-1 was included during the glucose stimulus . In conclusion , O95263 is expressed in a small number of tissues unrelated to glucose or fat metabolism . We propose that O95263 , an DB07954 -insensitive DB02527 -specific phosphodiesterase , could prove a novel target for enhanced insulin response , affecting a specific pool of DB02527 involved in the control of insulin granule trafficking and exocytosis . Finally , we discuss evidence for functional compartmentation of DB02527 in pancreatic beta-cells . Selected P21554 polymorphisms and hyperandrogenemia as well as fat mass and fat distribution in women with polycystic ovary syndrome . The endocannabinoid system is postulated to play an important role in the etiology of obesity , insulin resistance , fat distribution and metabolic disorders . P01308 resistance associated with abdominal obesity plays a leading role in the etiology of hyperandrogenism and other clinical features of the polycystic ovary syndrome ( PCOS ) . A total of 174 women 16-38 years old , diagnosed with PCOS according to the Rotterdam criteria are recruited . Control group consisted of 125 healthy women 18-45 years old . Medical history , physical examination , anthropometric parameters and metabolic parameters were carried out . Six P21554 gene polymorphisms were diagnosed . We observed a significantly three times higher risk of GG genotype in the polymorphism rs12720071 in women with PCOS versus the control group ( p = 0.0344 , OR = 3.01 ) . A similar , significant 8-fold higher risk ( p = 0.0176 , OR = 8.81 ) was demonstrated for genotype CC polymorphism rs806368 associated with PCOS . We observed a 3.6-fold increased risk of hyperandrogenemia ( free androgen index - FAI > 7 ) in patients with GG genotype in the rs12720071 polymorphism and AA genotype in the polymorphism rs1049353 ( OR = 2.7 ) . Our study may indicate a role of the endocannabinoid system in the occurrence of a specific hyperandrogenemia phenotype of PCOS . 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 . 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 .

MH_train_1108

MH_train_1108

MH_train_1108

interacts_with DB08865?

multiple_choice

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 . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . 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 . (-)- DB03823 -3-gallate ( EGCG ) inhibits P14210 -induced invasion and metastasis in hypopharyngeal carcinoma cells . P14210 ( P14210 ) has recently attracted a considerable amount of attention as a stromal-derived mediator in tumor-stromal interactions , particularly because of its close involvement in cancer invasion and metastasis , and DB03823 -3-gallate ( EGCG ) can modulate the cell signaling associated with angiogenesis , metastasis , and migration of cancer cells . In the present study , we have investigated the effects of P14210 on invasion and metastasis of hypopharyngeal carcinoma cells and the effect of EGCG on blocking P14210 -induced invasion and metastasis in these cells . We found that P14210 promoted the autophosphorylation of c- DB00134 , P08581 , and that P14210 -induced proliferation , colony dispersion , migration and invasion of tumors . We also observed that P14210 enhanced the activity of matrix metalloproteinase ( MMP ) -9 and urokinase-type plasminogen activator ( uPA ) in hypopharyngeal carcinoma cells . In addition , P14210 -induced the activation of Akt and Erk pathway as a downstreaming pathway of invasion . On the other hand , EGCG at physiologically relevant concentration ( 1 microM ) suppressed P14210 -induced tumor motility and P14780 and uPA activities , and the suppression of Akt and Erk pathway by EGCG was one of the downstream mechanisms to facilitate EGCG-induced anti-invasion effects . These results suggest that EGCG may serve as a therapeutic agent to inhibit P14210 -induced invasion in hypopharyngeal carcinoma patients . 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 . Stromal cells of endometrial carcinoma promotes proliferation of epithelial cells through the P14210 /c- DB00134 /Akt signaling pathway . Tumor microenvironment participates in the endometrial carcinoma pathogenesis . This study focuses on the interaction between endometrial cancer stromal cells and epithelial cells from normal endometrium tissue using in vitro transwell coculture system and in vivo xenograft model . We demonstrate that cancer interstitial ( CI ) cells stimulate normal epithelial ( NE ) cell proliferation . Tumor xenograft model confirmed the pro-proliferative effect of CI cells on epithelial cell growth . Tumor suppressor P60484 was reduced , and oncogene K-ras was increased in epithelial cells cocultured with CI cells . Moreover , we observed increased expression of hepatocyte growth factor ( P14210 ) in CI cells and tumor xenografts derived from the coculturing system . Higher P14210 secretion activated Akt signaling pathway , which was reversed by P08581 inhibitor ( crizotinib ) . These results demonstrate that endometrial carcinoma stromal cells stimulate epithelial cell proliferation via the P14210 /c- DB00134 /Akt signaling pathway . 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 . 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 . 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 . A Hepatocyte Growth Factor ( P14210 ) /receptor autocrine loop regulates constitutive self-renewal of human periodontal ligament cells but reduces sensitivity to exogenous P14210 . BACKGROUND : In addition to its prominent role in liver regeneration , hepatocyte growth factor ( P14210 ) is now generally thought to be produced by mesenchymal cells to promote the regeneration of epithelial tissue by a paracrine mechanism . However , it is not known how or if P14210 could be involved in the regeneration of periodontal tissues . The purpose of this study was to characterize the ability of normal human periodontal ligament ( PDL ) cells to produce or respond to P14210 . METHODS : PDL cells derived from healthy young volunteers were used from passages four through 10 . P14210 receptors were detected both by immunocytochemical staining and Western-blotting analysis . Both DNA synthesis ( by bromo-deoxyuridine [ BrdU ] -incorporation ) and secreted P14210 were quantified by enzyme-linked immunosorbent assays . Mitogen-activated protein kinase ( MAPK ) phosphorylation was also analyzed by Western blot . RESULTS : Despite the immunocytochemical demonstration of P08581 protein in the cytoplasm and on the plasma membrane of PDL cells , exogenous recombinant human P14210 did not exert the mitogenic effects expected . As reported for other mesenchymal cells , PDL cells were found to secrete P14210 . Treatments with neutralizing anti- P14210 antibody significantly suppressed constitutive PDL cell proliferation and sustained the receptor protein at higher levels than in non-treated cells . Under these conditions , exogenous P14210 rapidly phosphorylated extracellular signal-regulated kinase ( P29323 ) , an action linked to the cell proliferation and downregulation of cell-surface receptors . CONCLUSIONS : Unlike other known mesenchymal or epithelial cells , these findings suggest that normal PDL cells from young donors possess a constitutive P14210 /receptor autocrine loop that normally regulates their replacement self-proliferation but reduces sensitivity to exogenously applied P14210 by acute receptor downregulation . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 .

MH_train_1109

MH_train_1109

MH_train_1109

interacts_with DB00470?

multiple_choice

Flow cytometric measurement of intracellular Th1 and Th2 cytokine production by human villous and extravillous cytotrophoblast . A wide variety of cytokines are present at the maternal-fetal interface , but the extreme cellular complexity of the placenta has made it difficult to determine which cytokines are produced by which cells . Hence novel flow cytometric methods have been applied to determine intracellular cytokine production by specific cell-types in placental cell suspensions . Cell suspensions were prepared from first and third trimester chorionic villi and third trimester amniochorion by enzymatic digestion and Percoll density gradient centrifugation . After overnight incubation in the presence of monensin , cells were fixed , permeabilized and labelled with antibodies for villous cytotrophoblast ( cytokeratin+ , MHC class I- ) , extravillous cytotrophoblast ( cytokeratin+ , MHC class 1+ ) and leucocytes ( P08575 + ) . These cell types were further characterized by their expression of P00533 ( proliferative cytotrophoblast ) and c-erbB2 ( invasive cytotrophoblast ) . Production of P05112 , P22301 , P01375 , P01579 and IL-12 was determined by simultaneous labelling with the appropriate monoclonal antibodies . Only P05112 was detected consistently in all samples of cytotrophoblast . P22301 was not detected but P22301 mRNA was demonstrated in third trimester chorionic villus digests by RT-PCR . Although P05112 secretion has not been demonstrated , these data suggest that , in vivo there may be a " Th2 type cytokine bias " orchestrated by the trophoblast . It is proposed that other cytokines ( including P22301 and P01375 ) are produced by decidual leukocytes , and not cytotrophoblast , at the maternal-fetal interface . 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 . DB00470 excites rat VTA dopamine neurons through activation of cannabinoid P21554 but not opioid receptors . Behavioral , biochemical and recent electrophysiological data have increasingly implicated the involvement of dopamine in the central actions of cannabinoid compounds . However , the site and mechanism by which cannabinoids stimulate dopamine systems has been somewhat controversial . Central opioid systems have also been suggested to play a role in some cannabinoid-induced behaviors as evidenced by their attenuation in the presence of the opioid antagonist naloxone . However , recent studies using the cannabinoid receptor-selective antagonist SR141716A suggest that the central actions of psychoactive cannabinoids are mediated principally through activation of P21554 receptors . Using single cell electrophysiological recordings in the rat we assessed the effects of both SR141716A and naloxone on delta9-tetrahydrocannabinol ( THC ) -induced activation of ventral tegmental dopamine neurons . While dopamine cell firing was dose-dependently increased following cumulative dosing with delta9-THC it was partially or completely inhibited following pretreatment with 0.5 and 2 mg/kg SR141716A , respectively . However , 1 and 10 mg/kg naloxone failed to alter the response to delta9-THC . These data provide the first evidence that delta9-THC-induced changes in mesolimbic dopamine neuronal activity are mediated by the P21554 cannabinoid receptor , but a causal link for the involvement of opioid systems could not be established . Gene network analysis leads to functional validation of pathways linked to cancer cell growth and survival . Hepatocellular carcinoma ( HCC ) represents one of the most frequently diagnosed human cancers ; however , there are currently few treatment alternatives to surgical resection . In this study we performed bioinformatic analysis of previously published transcriptomic data in order to characterize liver specific networks , including biological functions , signaling pathways and transcription factors , potentially dysregulated in HCC . By incorporating specific signaling inhibitors into real-time proliferation assays using HepG2 cells , we then validated these in silico results . We found that G protein subunits Gi/G0 , protein kinase C , Mek1/2 , and Erk1/2 ( P42/44 ) , P23458 , Q07869 and NFκB p65 subunit were the major signaling molecules required for survival and proliferation of human HCC cell lines . We also found that these pathways regulate the expression of key hepatic transcription factors involved in cell differentiation , such as P49715 , P18146 , Q08050 and PPARs . By combining bioinformatic and functional analyses , major signaling pathways related to tumorigenicity in HCC are revealed , thereby elucidating potential targets for drug therapies . Delta9-tetrahydrocannabinol is a full agonist at P21554 receptors on GABA neuron axon terminals in the hippocampus . Marijuana impairs learning and memory through actions of its psychoactive constituent , DB00470 ( Delta(9)-THC ) , in the hippocampus , through activation of cannabinoid P21554 receptors ( CB1R ) . CB1Rs are found on glutamate and GABA neuron axon terminals in the hippocampus where they control neurotransmitter release . Previous studies suggest that Delta(9)-THC is a partial agonist of CB1Rs on glutamate axon terminals in the hippocampus , whereas its effects on GABA terminals have not been described . Using whole-cell electrophysiology in brain slices from C57BL6/J mice , we examined Delta(9)-THC effects on synaptic GABA IPSCs and postsynaptic GABA currents elicited by laser-induced photo-uncaging ( photolysis ) of alpha-carboxy-2-nitrobenzyl ( P63098 ) caged GABA . Despite robust inhibition of synaptic IPSCs in wildtype mice by the full synthetic agonist WIN55,212-2 , using a Tween-80 and DB01093 vehicle , Delta(9)-THC had no effects on IPSCs in this , or in a low concentration of another vehicle , randomly-methylated beta-cyclodextrin ( RAMEB , 0.023 % ) . However , IPSCs were inhibited by Delta(9)-THC in 0.1 % RAMEB , but not in neurons from CB1R knockout mice . Whereas Delta(9)-THC did not affect photolysis-evoked GABA currents , these responses were prolonged by a GABA uptake inhibitor . Concentration-response curves revealed that the maximal effects of Delta(9)-THC and WIN55,212-2 were similar , indicating that Delta(9)-THC is a full agonist at CB1Rs on GABA axon terminals . These results suggest that Delta(9)-THC inhibits GABA release , but does not directly alter GABA(A) receptors or GABA uptake in the hippocampus . Furthermore , full agonist effects of Delta(9)-THC on IPSCs likely result from a much higher expression of CB1Rs on GABA versus glutamate axon terminals in the hippocampus . 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 . Association study of a cannabinoid receptor gene ( P21554 ) polymorphism and schizophrenia . Cannabis can induce schizophrenic-like symptoms in healthy individuals . A principal active ingredient of cannabis , DB00470 , acts in the brain on a specific receptor , termed the cannabinoid receptor 1 ( P21554 ) . The human gene for P21554 is mapped to chromosome 6q14-15 , and linkage studies have produced evidence for a schizophrenia-susceptibility locus in this region . To explore a possible role for P21554 in the pathogenesis of schizophrenic disorders , we used an association study to genotype the P21554 polymorphism for 127 schizophrenic patients and 146 control subjects . The results demonstrate no association between P21554 genotypes and schizophrenic disorders ( P = 0.409 ) , with these negative findings suggesting that , for Chinese populations , the ( P01009 )n triplet repeat in the promoter region of the P21554 gene is not directly involved in the pathogenesis of schizophrenic disorders . 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 . ∆(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 . Cytoplasmic and nuclear estrogen binding capacity in the rat uterus during treatment with danazol and testosterone . DB01406 , testosterone and dihydrotestosterone ( DB02901 ) were tested as competitors for estrogen receptors on immature rat uterus cytosol . No competitive binding could be demonstrated for any of these steroids . After that , prepubertal Wistar rats were exposed to danazol , testosterone or propylene glycol ( control ) for 3 days or 17 days . After the appropriate exposure to medication , the animals were killed . Both danazol and testosterone appeared to be uterotropic after 3 days of treatment , although the increase in the uterine weight was significant only in the danazol-treated group ( p less than 0.05 ) . This effect was lost after 17 days of treatment . P03372 binding assays were done on the cytosolic and nuclear fractions of the homogenized uterine tissue of each group . The estrogen binding capacity of cytosols was increased in both the danazol ( p less than 0.05 ) and the testosterone ( p less than 0.01 ) groups after 3 days of treatment . A parallel increase was found in the nuclear fraction of both groups . After 17 days of treatment , the comparison between the 3 groups showed no differences in the cytosolic or nuclear estrogen binding capacity . The information provided by this study suggests that some effects of danazol may be due to an androgenic action and that may be associated to increases in the free fraction of testosterone . [ Endometrial cancer in young women -- clinical and molecular aspects ] . OBJECTIVES : The aim the study was to compare two groups of endometrial cancer patients ( below and above 45 years of age ) in the aspect of clinicopathological and molecular data . MATERIAL AND METHODS : The study encompassed 456 primary tumour samples retrospectively collected from a cohort of endometrial cancer patients , primarily treated by surgery Molecular analysis covered : copy number variations of 10 genes ( P11388 , P00533 , P04626 , P21860 , Q15303 , MYC , P24385 , P03372 , P42336 , O60216 ) analyzed by quantitative PCR ; mRNA expression of 6 genes ( Q13296 , RAD27 , Q01196 , O95863 , O43623 , O43490 ) analyzed with the use of reverse transcription quantitative PCR ; protein expression analysis of 8 markers ( P06401 , P03372 ; P00533 , P04626 , P21860 , Q15303 , P11388 , pAKT1 ) performed with the use of immunohistochemistry . RESULTS : The younger group of patients was characterized by less frequent hypertension ( p < 0.00007 ) , less frequent myometrial infiltration ( p=0.002 ) and longer overall survival ( p=0.003 ) . Apart from O60216 gene aberrations , which were more frequent in younger patients ( p=0.02 ) , the study revealed no statistically significant differences between the groups . CONCLUSIONS : The study showed no molecular differences in the profile of younger and older endometrial cancer patients . Data on both the prognostic and predictive significance of O60216 in endometrial cancer are still insufficient . The clinical profile of younger patients with endometrial carcinoma was slightly better when compared to elderly patients . Younger patients were characterized by longer overall survival . (-)-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 . Analysis of breast cancer related gene expression using natural splines and the Cox proportional hazard model to identify prognostic associations . Many studies correlating gene expression data to clinical parameters assume a linear increase or decrease of the clinical parameter under investigation with the expression of a gene . We have studied genes encoding important breast cancer-related proteins using a model for survival-type data that is based on natural splines and the Cox proportional hazard model , thereby removing the linearity assumption . Expression data of 16 genes were studied in relation to metastasis-free probability in a cohort of 295 consecutive breast cancer patients treated at The Netherlands Cancer Institute . The independent predictive power for disease outcome of the 16 individual genes was tested in a multivariable model with known clinical and pathological risk factors . There is a linear relationship between increasing expression and a higher or lower hazard for distant metastasis for P03372 , Q15303 , P15692 , O96020 , Q15910 , and Q96NZ9 ; for P04626 , P21860 , P24385 , P24864 , O75530 , P61073 , P32248 , P48061 , and P05121 there is no clear increase or decrease ; and for P00533 there seems to be a non-linear relation . Multivariable analysis showed that the 70-gene prognosis profile outperforms all the other variables in the model ( hazard-rate 5.4 , 95 % CI 2.5-11.7 ; P = 0.000018 ) . P00533 -expression seems to have a non-linear relation with disease outcome , indicating that lower but also higher expression of P00533 are associated with worse outcome compared to intermediate expression levels ; the other genes show no or a linear relation . P60568 suppression by 2-arachidonyl glycerol is mediated through peroxisome proliferator-activated receptor gamma independently of cannabinoid receptors 1 and 2 . 2-Arachidonyl glycerol ( 2-AG ) is an endogenous arachidonic acid derivative that binds cannabinoid receptors P21554 and CB2 and is hence termed an endocannabinoid . 2-AG also modulates a variety of immunological responses , including expression of the autocrine/paracrine T cell growth factor interleukin ( IL ) -2 . The objective of the present studies was to determine the mechanism responsible for P60568 suppression by 2-AG . Because of the labile properties of 2-AG , 2-AG ether , a nonhydrolyzable analog of 2-AG , was also used . Both 2-AG and 2-AG ether suppressed P60568 expression independently of P21554 and CB2 , as demonstrated in leukocytes derived from P21554 /CB2-null mice . Moreover , we demonstrated that both 2-AG and 2-AG ether treatment activated peroxisome proliferator-activated receptor gamma ( PPARgamma ) , as evidenced by forced differentiation of 3T3- Q9NUQ9 cells into adipocytes , induction of aP2 mRNA levels , and activation of a PPARgamma-specific luciferase reporter in transiently transfected 3T3- Q9NUQ9 cells . Consequently , the putative role of PPARgamma in P60568 suppression by 2-AG and 2-AG ether was examined in Jurkat T cells . Concordant with PPARgamma involvement , the PPARgamma-specific antagonist 2-chloro-5-nitro-N-(4-pyridyl)-benzamide ( T0070907 ) blocked 2-AG- and 2-AG ether-mediated P60568 suppression . Likewise , 2-AG suppressed the transcriptional activity of two transcription factors crucial for P60568 expression , nuclear factor of activated T cells and nuclear factor kappaB , in the absence but not in the presence of T0070907 . 2-AG treatment also induced PPARgamma binding to a Q07869 response element in activated Jurkat T cells . Together , the aforementioned studies identify PPARgamma as a novel intracellular target of 2-AG , which mediates the suppression of P60568 by 2-AG in a manner that is independent of P21554 and/or CB2 . [ Neuropsychopharmacology of DB00470 ] . Today , the main route of introduction of tetrahydrocannabinol ( THC ) , the main active substance of cannabis , into the human body is via the lungs , from smokes produced by combustion of a haschich-tobacco mixture . The use of a water pipe ( nargileh-like ) intensifies its fast supply to the body . THC reaches the brain easily where it stimulates P21554 receptors ; their ubiquity underlies a wide variety of effects . THC disappears from extracellular spaces by dissolving in lipid rich membranes , and not by excretion from the body . This is followed by a slow release , leading to long lasting effects originating from brain areas containing a large proportion of spare receptors ( " reserve receptors " ) . Far from mimicking the effects of endocannabinoids , THC caricatures and disturbs them . It induces both psychical and physical dependencies , but the perception of withdrawal is weak on account of its very slow elimination . THC disturbs cognition . Acutely , it develops anxiolytic- and antidepressant-like effects , which causes a lot of users to abuse THC , thus leading to a tolerance ( desensitization of P21554 receptors ) making anxiety and depression to reappear more intensely than originally . THC has close relationships with schizophrenia . It incites to tobacco , alcohol and heroine abuses . 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 . P34972 undergoes Rab5-mediated internalization and recycles via a Rab11-dependent pathway . P34972 ( CB2 ) is a GPCR highly expressed on the surface of cells of the immune system , supporting its role in immunomodulation . This study has investigated the trafficking properties of this receptor when stably expressed by P29320 -293 cells . As previously reported , cell surface CB2 rapidly internalized upon exposure to agonist . Direct evidence of CB2 recycling was observed upon competitive removal of the stimulating agonist by inverse agonist . CB2 also underwent slow constitutive internalization when agonist was absent and was up-regulated in the presence of inverse agonist . Co-expression of CB2 and dominant negative Rab5 resulted in a significantly reduced capacity for receptors to internalize with no effect on recycling of the internalized receptors . Conversely , co-expression with dominant negative Rab11 did not alter the ability of CB2 to internalize but did impair their ability to return to the cell surface . Co-expression of wild-type , dominant negative or constitutively active Rab4 with CB2 did not alter basal surface expression , extent of internalization , or extent of recycling . These results suggest that Rab5 is involved in CB2 endocytosis and that internalized receptors are recycled via a Rab11 associated pathway rather than the rapid Rab4 associated pathway . This report provides the first comprehensive description of CB2 internalization and recycling to date . [ Characteristics of abnormal behavior induced by delta 9-tetrahydrocannabinol in rats ] . delta 9- DB00470 ( THC ) , one of the active compounds of marihuana , is known to induce drug dependence and tolerance , and its action is weaker than those of other abused drugs in humans and animals . Acute effects of THC , " high " , " irritable " and " cognitive deficits " are more important than the drug dependence and tolerance . For this reason , we examined characteristics of abnormal behavior such as catalepsy-like immobilization , aggressive behavior including irritable aggression and muricide , and spatial cognition impairment induced by acute and chronic treatments of THC in rats . The catalepsy-like immobilization is related to a decrease in catecholaminergic and serotonergic neurons in the nucleus accumbens and amygdaloid nucleus and thus serves as a useful model for amotivational syndrome , one of cannabis psychoses . In aggressive behavior , muricide was determined by the housing condition . Muricide was induced if the rat was placed under an isolated housing condition within the period of the effect of single injection of THC . The behavioral change resembles exacerbation and flashback in humans . Spatial cognition is impaired by the interaction between cannabinoid ( P21554 ) and 5-HT2 receptor in the dorsal raphe-hippocampal serotonergic neurons . Thus the abnormal behavior induced by THC can be a useful model for investigating mental function in humans and new drugs for the treatment of mental disorders . Boosting the sampling efficiency of q-Ball imaging using multiple wavevector fusion . q-Ball imaging ( QBI ) is a high-angular-resolution diffusion imaging ( HARDI ) method that is capable of resolving complex , subvoxel white matter ( WM ) architecture . QBI requires time-intensive sampling of the diffusion signal and large diffusion wavevectors . Here we describe a reconstruction scheme for QBI , termed multiple wavevector fusion ( MWF ) , that substantially boosts the sampling efficiency and signal-to-noise ratio ( SNR ) of QBI . The MWF reconstruction operates by nonlinearly fusing the diffusion signal from separate low and high wavevector acquisitions . The combination of wavevectors provides the benefits of the high SNR of the low wavevector signal and the high angular contrast-to-noise ratio ( P21554 ) and peak separation of the high wavevector signal . The MWF procedure provides a framework for combining diffusion tensor imaging ( DTI ) and QBI . Numerical simulations show that MWF of DTI and QBI provides a more accurate estimate of the diffusion orientation distribution function ( O14788 ) than QBI alone . The accuracy improvement can be translated into an efficiency gain of 274-377 % . An intravoxel peak connectivity metric ( IPCM ) is presented that calculates the peak connectivity between an O14788 and its neighboring voxels . In human WM , MWF reveals more detailed WM architecture than QBI as measured by the IPCM for all sampling schemes presented . 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 . 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 . 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 . [ 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 . 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 . (+)- 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 . 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 . Chronic DB00470 treatment increases DB02527 levels and DB02527 -dependent protein kinase activity in some rat brain regions . When Delta(9)-tetrahydrocannabinol ( Delta(9)-THC,15 mg/kg ) was injected intraperitoneally twice a day for 6 days , tolerance to its analgesic effect appeared to be complete . Chronic exposure to Delta(9)-THC caused a significant reduction in P21554 receptor binding in all brain areas that contain this receptor . Cannabinoid receptor density was markedly reduced in the cerebellum ( 52 % ) , hippocampus ( 40 % ) and globus pallidum ( 47 % ) compared to 30 % in the cortex and striatum . Chronic exposure enhanced the DB02527 pathway , as shown by the significant increase of DB02527 levels and PKA activity in the areas with receptor down-regulation ( cerebellum , striatum and cortex ) . We propose that the increase in DB02527 cascade is part of the biochemical basis of cannabinoid tolerance . Δ(9)- DB00470 acts as a partial agonist/antagonist in mice . Δ- DB00470 ( THC ) has been characterized as a partial agonist at cannabinoid P21554 receptors in vitro ; however , it often produces the same maximum effects in vivo as other cannabinoid agonists . This study was carried out to determine whether THC would antagonize the hypothermic effects of another cannabinoid agonist , AM2389 , in mice . Male mice were injected with 1-100 mg/kg THC , 0.01-0.1 mg/kg AM2389 , or a combination of 30 mg/kg THC and 0.1-1.0 mg/kg AM2389 , and rectal temperature was recorded for up to 12 h after injection . THC reduced the temperature by 5.6°C at a dose of 30 mg/kg ; further increases in the dose did not produce larger effects , indicating a plateau in the THC dose-effect function . AM2389 reduced temperature by 9.0°C at a dose of 0.1 mg/kg . One hour pretreatment with 30 mg/kg THC attenuated the hypothermic effects of 0.1 mg/kg AM2389 ; a 10-fold higher dose , 1.0 mg/kg AM2389 , was required to further decrease temperature , reflecting a five-fold rightward shift of the lower portion of the AM2389 dose-effect function following THC pretreatment . These results indicate that , in an assay of mouse hypothermia , THC exerts both agonist and antagonist effects following acute administration , and mark the first demonstration of partial agonist/antagonist effects of THC in vivo . 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 . 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 . 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 . 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 ) . 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 . [ 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 . 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 . 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 . AB-CHMINACA , AB-PINACA , and FUBIMINA : Affinity and Potency of Novel Synthetic Cannabinoids in Producing Δ9- DB00470 -Like Effects in Mice . Diversion of synthetic cannabinoids for abuse began in the early 2000s . Despite legislation banning compounds currently on the drug market , illicit manufacturers continue to release new compounds for recreational use . This study examined new synthetic cannabinoids , AB-CHMINACA ( N-[1-amino-3-methyl-oxobutan-2-yl]-1-[cyclohexylmethyl]-1H-indazole-3-carboxamide ) , AB-PINACA [ N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-pentyl-1H-indazole-3-carboxamide ] , and FUBIMINA [ ( 1-(5-fluoropentyl)-1H-benzo[d]imadazol-2-yl ) (naphthalen-1-yl)methanone ] , with the hypothesis that these compounds , like those before them , would be highly susceptible to abuse . Cannabinoids were examined in vitro for binding and activation of P21554 receptors , and in vivo for pharmacological effects in mice and in Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) discrimination . AB-CHMINACA , AB-PINACA , and FUBIMINA bound to and activated P21554 and CB2 receptors , and produced locomotor suppression , antinociception , hypothermia , and catalepsy . Furthermore , these compounds , along with JWH-018 [ 1-pentyl-3-(1-naphthoyl)indole ] , Q13515 ,497 [ rel-5-(1,1-dimethylheptyl)-2- [ ( 1R,3S ) -3-hydroxycyclohexyl ] -phenol ] , and WIN55,212-2 ( [ ( 3R ) -2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl ] -1-naphthalenyl-methanone , monomethanesulfonate ) , substituted for Δ(9)-THC in Δ(9)-THC discrimination . Rank order of potency correlated with P21554 receptor-binding affinity , and all three compounds were full agonists in [(35)S]GTPγS binding , as compared with the partial agonist Δ(9)-THC . Indeed , AB-CHMINACA and AB-PINACA exhibited higher efficacy than most known full agonists of the P21554 receptor . Preliminary analysis of urinary metabolites of the compounds revealed the expected hydroxylation . AB-PINACA and AB-CHMINACA are of potential interest as research tools due to their unique chemical structures and high P21554 receptor efficacies . Further studies on these chemicals are likely to include research on understanding cannabinoid receptors and other components of the endocannabinoid system that underlie the abuse of synthetic cannabinoids . 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 . Cannabinoid agonists and antagonists modulate lithium-induced conditioned gaping in rats . Considerable evidence indicates that conditioned gaping in rats reflects nausea in this species that does not vomit . A series of experiments evaluated the potential of psychoactive cannabinoid agonists , DB00470 and HU-210 , and non-psychoactive cannabinoids , DB09061 ( DB09061 ) and its dimethylheptyl homolog ( DB09061 -dmh ) , to interfere with the establishment and the expression of conditioned gaping in rats . All agents attenuated both the establishment and the expression of conditioned gaping . Furthermore , the P21554 antagonist , SR-141716 , reversed the suppressive effect of HU-210 on conditioned gaping . Finally , SR-141716 potentiated lithium-induced conditioned gaping , suggesting that the endogenous cannabinoid system plays a role in the control of nausea . 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 . 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 . 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 . 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 . P34972 -mediated attenuation of P61073 -tropic HIV infection in primary P01730 + T cells . Agents that activate cannabinoid receptor pathways have been tested as treatments for cachexia , nausea or neuropathic pain in HIV-1/AIDS patients . The cannabinoid receptors ( CB(1)R and CB(2)R ) and the HIV-1 co-receptors , P51681 and P61073 , all signal via Gαi-coupled pathways . We hypothesized that drugs targeting cannabinoid receptors modulate chemokine co-receptor function and regulate HIV-1 infectivity . We found that agonism of CB(2)R , but not CB(1)R , reduced infection in primary P01730 + T cells following cell-free and cell-to-cell transmission of P61073 -tropic virus . As this change in viral permissiveness was most pronounced in unstimulated T cells , we investigated the effect of CB(2)R agonism on to P61073 -induced signaling following binding of chemokine or virus to the co-receptor . We found that CB(2)R agonism decreased P61073 -activation mediated G-protein activity and MAPK phosphorylation . Furthermore , CB(2)R agonism altered the cytoskeletal architecture of resting P01730 + T cells by decreasing F-actin levels . Our findings suggest that CB(2)R activation in P01730 + T cells can inhibit actin reorganization and impair productive infection following cell-free or cell-associated viral acquisition of P61073 -tropic HIV-1 in resting cells . Therefore , the clinical use of CB(2)R agonists in the treatment of AIDS symptoms may also exert beneficial adjunctive antiviral effects against P61073 -tropic viruses in late stages of HIV-1 infection . 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 . 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 . 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 . 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 . 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 . The molecular basis of the cooperation between P01133 , FGF and eCB receptors in the regulation of neural stem cell function . Adult neurogenesis relies on P01133 and FGF receptor ( P00533 /FGFR ) function and endocannabinoid ( eCB ) signalling . Here we have used a neural stem cell ( NSC ) line to determine how these systems cooperate to regulate neurogenesis . The results show the P00533 to be solely responsible for maintaining PI3K activation explaining its dominant role in promoting NSC survival . The P00533 and FGFR synergistically regulate the P29323 /MAPK pathway , and this explains the requirement for both for optimal cell proliferation . The eCB receptors did not contribute to activation of the PI3K or P29323 /MAPK pathways , highlighting the importance of another major proliferation pathway . The P00533 plays the dominant role in maintaining the transcriptome , with significant changes in the expression of over 3500 transcripts seen within hours of inhibition or activation of this receptor . The FGFR has a more modest effect on transcription with evidence for nodal integration with P00533 signalling at the level of the P29323 /MAPK pathway . A common set of transcripts are regulated by the P21554 and CB2 receptors , with cooperation between these receptors and the P00533 apparent in the regulation of a pool of transcripts , most likely representing signal integration downstream from an as yet to be identified node . Finally , a first level molecular analysis of the transcriptional response shows regulation of a number of key growth factors , growth factor receptors and GPCRs to be under the control of the P00533 . 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 . 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 . 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 . 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 . 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 . Stimulation of host bone marrow stromal cells by sympathetic nerves promotes breast cancer bone metastasis in mice . Bone and lung metastases are responsible for the majority of deaths in patients with breast cancer . Following treatment of the primary cancer , emotional and psychosocial factors within this population precipitate time to recurrence and death , however the underlying mechanism(s) remain unclear . Using a mouse model of bone metastasis , we provide experimental evidence that activation of the sympathetic nervous system , which is one of many pathophysiological consequences of severe stress and depression , promotes MDA-231 breast cancer cell colonization of bone via a neurohormonal effect on the host bone marrow stroma . We demonstrate that induction of O14788 expression in bone marrow osteoblasts , following β2AR stimulation , increases the migration of metastatic MDA-231 cells in vitro , independently of P48061 - P61073 signaling . We also show that the stimulatory effect of endogenous ( chronic stress ) or pharmacologic sympathetic activation on breast cancer bone metastasis in vivo can be blocked with the β-blocker propranolol , and by knockdown of Q9Y6Q6 expression in MDA-231 cells . These findings indicate that O14788 promotes breast cancer cell metastasis to bone via its pro-migratory effect on breast cancer cells , independently of its effect on bone turnover . The emerging clinical implication , supported by recent epidemiological studies , is that βAR-blockers and drugs interfering with O14788 signaling , such as DB06643 , could increase patient survival if used as adjuvant therapy to inhibit both the early colonization of bone by metastatic breast cancer cells and the initiation of the " vicious cycle " of bone destruction induced by these cells . 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 . 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 . Genetic variation in the adiponectin receptor 2 ( Q86V24 ) gene is associated with coronary artery disease and increased Q86V24 expression in peripheral monocytes . BACKGROUND : Q15848 is an adipose tissue secreted protein known for its insulin sensitising and anti-atherogenic actions . To this date two adiponectin receptors have been discovered , adiponectin receptor 1 ( Q96A54 ) and adiponectin receptor 2 ( Q86V24 ) . The aim of this study was to investigate the association of Q86V24 gene variations with coronary artery disease ( CAD ) . METHODS : Eight common single nucleotide polymorphisms ( SNPs ) spanning the entire Q86V24 locus were chosen to perform association studies with anthropometric and metabolic parameters in a Greek population . They were classified as either CAD ( stenosis > 50 % in at least one main vessel ) or non-CAD individuals in accordance with coronary angiography data.Genotyping was performed using a microsphere-based suspension array and the Allele Specific Primer Extension ( ASPE ) method . Expression of Q86V24 protein and mRNA in circulating P08571 + monocytes were determined using flow cytometry and real time Polymerase Chain Reaction assays respectively . RESULTS : There was a significant difference in the distribution of genotypes of polymorphism rs767870 of Q86V24 between CAD and non-CAD individuals ( p = 0.017 ) . Furthermore , heterozygotes of the rs767870 polymorphism had significantly lower Flow Mediated Dilatation ( FMD ) values , higher values of Intima-Media Thickness ( IMT ) and increased Q86V24 protein levels in peripheral monocytes , compared to homozygotes of the minor allele after adjustment for age , sex , waist to hip ratio and HOMA . CONCLUSIONS : Our findings suggest that variants of Q86V24 could be a determinant for atherosclerosis independent of insulin resistance status , possibly by affecting Q86V24 protein levels . 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 . Δ(9)- DB00470 treatment during human monocyte differentiation reduces macrophage susceptibility to HIV-1 infection . The major psychoactive component of marijuana , Δ(9)-tetrahydrocannabinol ( THC ) , also acts to suppress inflammatory responses . Receptors for THC , P21554 , CB2 , and Q9Y2T6 , are differentially expressed on multiple cell types including monocytes and macrophages , which are important modulators of inflammation in vivo and target cells for HIV-1 infection . Use of recreational and medicinal marijuana is increasing , but the consequences of marijuana exposure on HIV-1 infection are unclear . Ex vivo studies were designed to investigate effects on HIV-1 infection in macrophages exposed to THC during or following differentiation . THC treatment of primary human monocytes during differentiation reduced HIV-1 infection of subsequent macrophages by replication competent or single cycle P51681 using viruses . In contrast , treatment of macrophages with THC immediately prior to or continuously following HIV-1 exposure failed to alter infection . Specific receptor agonists indicated that the THC effect during monocyte differentiation was mediated primarily through CB2 . THC reduced the number of p24 positive cells with little to no effect on virus production per infected cell , while quantitation of intracellular viral gag pinpointed the THC effect to an early event in the viral life cycle . Cells treated during differentiation with THC displayed reduced expression of P08571 , CD16 , and Q86VB7 and donor dependent increases in mRNA expression of selected viral restriction factors , suggesting a fundamental alteration in phenotype . Ultimately , the mechanism of THC suppression of HIV-1 infection was traced to a reduction in cell surface HIV receptor ( P01730 , P51681 and P61073 ) expression that diminished entry efficiency . 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 . 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 . 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 .

MH_train_1110

MH_train_1110

MH_train_1110

interacts_with DB01045?

multiple_choice

[ Development of simplified and rapid detection assay for genetic polymorphisms influencing drug response and its clinical applications ] . Clinically important genetic polymorphisms influencing drug metabolism and drug response have typically been discovered on the basis of phenotypic differences among individuals from different populations . Routine genotyping before drug therapy may enable the identification of responders , nonresponders , or patients at increased risk of toxicity . Automated , high-throughput detecting methods for single-nucleotide polymorphisms ( SNPs ) are highly desirable in many clinical laboratories . The aim of this study is to develop a high-throughput genotyping method for detecting SNPs influencing drug response in the Japanese population . We have developed three real-time PCR assays for detecting SNPs in the human drug-metabolizing enzymes and drug targets . The assay for simultaneously detecting P11509 , P20813 , P11712 , P33260 , P33261 , P10635 , P05181 , P20815 , NAT2 , P51580 , Q12882 , P22309 , P05091 , P00325 , P08183 , P11597 , P12821 -1 , P07550 , P28223 , P49441 , P48061 , and mitochondrial DNA polymorphisms takes less than 1.5 h . With the clinical application of NAT2 genotyping , we found statistically significant difference between the incidence of adverse drug reactions ( ADRs ) and the NAT2 genotype . The incidence of the ADRs was significantly higher in the slow type than the in other two types , as 5 of the 6 patients were of the slowtype , and the other was the intermediatetype , while no patients of the rapidtype has developed any ADRs . Combination cytotoxic chemotherapy with cisplatin or doxorubicin and photodynamic therapy in murine tumors . This study was designed to evaluate the interaction of photodynamic therapy ( PDT ) and chemotherapy in an animal model . PDT is based on the interaction of hematoporphyrin derivative and red light of the appropriate wavelength ( 630 nm ) and intensity . Two tumor models were utilized : C3H/Km mice bearing the Q9HBH0 -1 tumor and BALB/c mice bearing the EMT-6 tumor . Tumor-bearing mice were treated with either cisplatin ( O60220 ) , doxorubicin ( P35318 ) , PDT , or a combination of drug and PDT . It was demonstrated that the Q9HBH0 -1 tumor was sensitive to O60220 and insensitive to both PDT and P35318 . There was no additional antitumor effect when either drug was combined with PDT . The EMT-6 tumor was moderately sensitive to PDT and mildly sensitive to both O60220 and P35318 . Although the addition of O60220 did not potentiate tumor destruction , the addition of P35318 significantly enhanced the effect of PDT ( P = .01 ) . The enhanced activity of the combination of PDT and P35318 appeared to be the result of increased activity of P35318 alone , when illuminated with red ( 630 nm ) light . This potentiation may be due to a photochemical process or may be secondary to the mild hyperthermia generated by illumination with the laser . This study demonstrates that PDT combined with cytotoxic chemotherapy is well tolerated in these animals and that certain combinations of PDT and chemotherapy may result in an enhanced tumoricidal effect . 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 . 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 . Polymorphisms of chemokine receptors and P29474 in Brazilian patients with sickle cell disease . Sickle cell disease ( O00767 ) is an inherited disorder that presents extremely variable clinical manifestations . For the past few decades , it has been approached as an inflammatory disorder , and several researchers have tried to determine the factors involved in such characteristic . In order to contribute to the identification of the genetic differences underlying this phenotypic diversity in O00767 , we proposed to study the distribution of polymorphic variants of the genes encoding the chemokine receptors P41597 and P51681 , as well as three polymorphisms in the NOS3 gene , in Brazilian O00767 patients . These genes are involved in the development of inflammatory immune reactions , a feature believed to be of extreme importance in O00767 pathology . Our results indicate that the polymorphisms studied here are not directly associated with severe clinical manifestations in O00767 patients . Nevertheless , we observed a tendency for the development of a severe clinical course in carriers of the variant alleles P41597 -64I and CCR5delta32 and in homozygotes for the -786C variant of the NOS3 gene . Further studies should be carried out in order to assess the role of such variants in the clinical picture of O00767 . Chemically synthesized SDF-1alpha analogue , N33A , is a potent chemotactic agent for P61073 / P61073 / P61073 -expressing human leukocytes . Stromal cell-derived factor ( SDF ) 1 is a potent chemoattractant for leukocytes through activation of the receptor P61073 / P61073 / P61073 , which is a fusion co-factor for the entry of T lymphocytotropic human immunodeficiency virus type 1 ( HIV-1 ) . This P61073 -mediated HIV-1 fusion can be inhibited by P48061 . Because of its importance in the study of immunity and AIDS , large scale production of P48061 is desirable . In addition to recombinant technology , chemical synthesis provides means by which biologically active proteins can be produced not only in large quantity but also with a variety of designed modifications . In this study , we investigated the binding and function of an SDF-1alpha analogue , N33A , synthesized by a newly developed native chemical ligation approach . Radioiodinated N33A showed high affinity binding to human monocytes , T lymphocytes , as well as neutrophils , and competed equally well with native recombinant SDF-1alpha for binding sites on leukocytes . N33A also showed equally potent chemoattractant activity as native recombinant SDF-1alpha for human leukocytes . Further study with P61073 / P61073 / P61073 transfected P29320 293 cells showed that N33A binds and induces directional migration of these cells in vitro . These results demonstrate that the chemically synthesized SDF-1alpha analogue , N33A , which can be produced rapidly in large quantity , possesses the same capacity as native SDF-1alpha to activate P61073 -expressing cells and will provide a valuable agent for research on the host immune response and AIDS . Comparative effects of cytokines on constitutive and inducible expression of the gene encoding for the cytochrome P450 3A6 isoenzyme in cultured rabbit hepatocytes : consequences on progesterone 6beta-hydroxylation . Cultured rabbit hepatocytes were used to compare the relative activities of cytokines to inhibit the constitutive or rifampicin ( Q9HBH0 ) -induced expression of the cytochrome P450 3A6 gene ( CYP3A6 ) . Human recombinant cytokines tested were interleukin-1beta ( IL-1beta ) ( 2 U/mL ) , interleukin-2 ( P60568 ) ( 5,000 U/mL ) and interferon-gamma ( P01579 ) ( 50 U/mL ) . Hepatocytes were cultured in the presence or absence of 25 microM Q9HBH0 for 24 hr , with or without cytokines alone or in combination . All these cytokines inhibited Q9HBH0 -induced P4503A6 expression without apparent cellular toxicity . By contrast , only P01579 treatment provided a significant decrease ( 41 % ) in the constitutive P4503A6 protein level . Moreover , cytokines differed in their ability to repress Q9HBH0 -dependent transcriptional induction of CYP3A6 : IL-1beta and P60568 were approximately equipotent , causing an almost 40-50 % suppression of CYP3A6 mRNA and protein levels , whereas P01579 exerted repressive effects only on P4503A6-related erythromycin N-demethylase activity and inducible protein expression . In fact , although strongly reducing P4503A6 protein content ( an approximate 70 % decrease ) , P01579 did not exhibit any influence on CYP3A6 mRNAs with the exception of its association with interleukins . All these results suggest that IL-1beta and P60568 mainly promote a transcriptional repression mechanism , given the absence of effect of these cytokines on the basal P4503A6 level , whereas P01579 exerts a post-transcriptional suppressive action on both induced and constitutive P4503A6 expression . Consequently , P4503A6-dependent progesterone 6beta-hydroxylase activity also presented a cytokine-specific pattern of inhibition , with a much greater sensitivity than P4503A6 immunoreactive protein to IL-1beta and P60568 + P01579 treatments . Thus , this study underlines the significant impact of inflammation on steroid metabolism . Sensitization to autoimmune hepatitis in group VIA calcium-independent phospholipase A2-null mice led to duodenal villous atrophy with apoptosis , goblet cell hyperplasia and leaked bile acids . Chronic bowel disease can co-exist with severe autoimmune hepatitis ( AIH ) in an absence of primary sclerosing cholangitis . Genetic background may contribute to this overlap syndrome . We previously have shown that the deficiency of iPLA2β causes an accumulation of hepatocyte apoptosis , and renders susceptibility for acute liver injury . We here tested whether AIH induction in iPLA2β-null mice could result in intestinal injury , and whether bile acid metabolism was altered . Control wild-type ( WT ) and female iPLA2β-null ( iPLA2β(-/-) ) mice were intravenously injected with 10mg/kg concanavalinA ( ConA ) or saline for 24h . ConA treatment of iPLA2β(-/-) mice caused massive liver injury with increased liver enzymes , fibrosis , and necrosis . While not affecting WT mice , ConA treatment of iPLA2β(-/-) mice caused severe duodenal villous atrophy concomitant with increased apoptosis , cell proliferation , globlet cell hyperplasia , and endotoxin leakage into portal vein indicating a disruption of intestinal barrier . With the greater extent than in WT mice , ConA treatment of iPLA2β(-/-) mice increased jejunal expression of innate response cytokines P08571 , P01375 -α , P05231 , and O14543 as well as chemokines P13500 and the P10147 receptor P51681 . iPLA2β deficiency in response to ConA-induced AIH caused a significant decrease in hepatic and biliary bile acids , and this was associated with suppression of hepatic Cyp7A1 , Ntcp and O95342 /Bsep and upregulation of intestinal Q96RI1 /FGF15 mRNA expression . The suppression of hepatic Ntcp expression together with the loss of intestinal barrier could account for the observed bile acid leakage into peripheral blood . Thus , enteropathy may result from acute AIH in a susceptible host such as iPLA2β deficiency . [ 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 . 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 . Self-assembly of P29320 cell-secreted ApoE particles resembles ApoE enrichment of lipoproteins as a ligand for the P01130 -related protein . Recent studies have shown that the lipidation and assembly state of apolipoprotein E ( apoE ) determine receptor recognition and amyloid-beta peptide ( Abeta ) binding . We previously demonstrated that apoE secreted by P29320 cells stably expressing apoE3 or apoE4 ( P29320 -apoE ) binds Abeta and inhibits Abeta-induced neurotoxicity by an isoform-specific process that requires apoE receptors . Here we characterized the structure of P29320 -apoE assemblies and determined their receptor binding specificity . By chromatography , P29320 -apoE elutes in high molecular mass fractions and is the size of plasma HDL , consistent with a multiprotein assembly . No lipid was associated with these apoE assemblies . Several methods for analyzing receptor binding indicate that P29320 -apoE is a ligand for low-density lipoprotein ( LDL ) receptor-related protein ( Q14764 ) but not the P01130 . This suggests that self-assembly of apoE may induce a functional conformation necessary for binding to Q14764 . Our results indicate that , in addition to lipid content , the assembly state of apoE influences Abeta binding and receptor recognition . 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 . 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 . Identification and organization of ribosomal protein genes of Escherichia coli carried by lambdafus2 transducing phage . We describe the isolation of lambdafus2 , which carries bacterial DNA from the str-spc region of the Escherichia coli chromosome . Genes for 27 ribosomal proteins were found on the genome of this transducing phage by identifying the ribosomal proteins whose synthesis was stimulated after infection of UV-irradiated bacteria . These were genes for S3 , S4 , S5 , S7 , S8 , S10 , S11 , P28222 , S13 , Q92748 , S17 , S19 , Q401N2 , Q96MH2 , Q9NP50 , Q15004 , Q9BTT4 , Q9H8H0 , Q96HJ3 , Q6PL18 , Q68CL5 , Q9NXF1 , L22 L23 , L24 , L29 , and L30 . Subsets of these genes were identified on the genomes of the phages lambdaspc1 , lambdaspc2-delta 9 , and lambdaspc2-delta16 , all of which carry parts of the bacterial DNA present on lambdafus2 . From the known structures of these phage genomes , it has been possible to determine the relative order of many of these genes on the lambdafus2 genome and thus on the E. coli chromosome . Our evidence also suggests that the genes for S3 , S17 , S19 , Q401N2 , Q9NP50 , Q6PL18 , L22 , L23 , and L29 are part of a single transcription unit . These results , along with the observations described in the previous and accompanying papers , indicate that the ribosomal protein genes on lambdafus2 are organized into at least four transcription units . Common variants near P42685 / Q03692 and P15692 are associated with advanced age-related macular degeneration . Despite significant progress in the identification of genetic loci for age-related macular degeneration ( AMD ) , not all of the heritability has been explained . To identify variants which contribute to the remaining genetic susceptibility , we performed the largest meta-analysis of genome-wide association studies to date for advanced AMD . We imputed 6 036 699 single-nucleotide polymorphisms with the 1000 Genomes Project reference genotypes on 2594 cases and 4134 controls with follow-up replication of top signals in 5640 cases and 52 174 controls . We identified two new common susceptibility alleles , rs1999930 on 6q21-q22.3 near P42685 / Q03692 [ odds ratio ( OR ) 0.87 ; P = 1.1 × 10(-8) ] and rs4711751 on 6p12 near P15692 ( OR 1.15 ; P = 8.7 × 10(-9) ) . In addition to the two novel loci , 10 previously reported loci in P0C7Q2 / Q92743 ( rs10490924 ) , P08603 ( rs1061170 , and rs1410996 ) , P00751 ( rs641153 ) , P01024 ( rs2230199 ) , P06681 ( rs9332739 ) , P05156 ( rs10033900 ) , P11150 ( rs10468017 ) , P35625 ( rs9621532 ) and P11597 ( rs3764261 ) were confirmed with genome-wide significant signals in this large study . Loci in the recently reported genes O95477 and P27658 were also detected with suggestive evidence of association with advanced AMD . The novel variants identified in this study suggest that angiogenesis ( P15692 ) and extracellular collagen matrix ( P42685 / Q03692 ) pathways contribute to the development of advanced AMD . 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 . 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) . 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 . 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 . The expression level of P21554 and CB2 receptors determines their efficacy at inducing apoptosis in astrocytomas . BACKGROUND : Cannabinoids represent unique compounds for treating tumors , including astrocytomas . Whether CB(1) and CB(2) receptors mediate this therapeutic effect is unclear . PRINCIPAL FINDINGS : We generated astrocytoma subclones that express set levels of CB(1) and CB(2) , and found that cannabinoids induce apoptosis only in cells expressing low levels of receptors that couple to P27361 /2 . In contrast , cannabinoids do not induce apoptosis in cells expressing high levels of receptors because these now also couple to the prosurvival signal AKT . Remarkably , cannabinoids applied at high concentration induce apoptosis in all subclones independently of CB(1) , CB(2) and AKT , but still through a mechanism involving P27361 /2 . SIGNIFICANCE : The high expression level of CB(1) and CB(2) receptors commonly found in malignant astrocytomas precludes the use of cannabinoids as therapeutics , unless AKT is concomitantly inhibited , or cannabinoids are applied at concentrations that bypass CB(1) and CB(2) receptors , yet still activate P27361 /2 . 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 . P49327 inhibition induces differential expression of genes involved in apoptosis and cell proliferation in ocular cancer cells . P49327 ( P49327 ) , a lipogenic multienzyme complex , is overexpressed in the ocular cancer , retinoblastoma , and is strongly correlated with tumor invasion . Dietary nutrients are reported to exert anticancer effects through inhibition of lipid metabolism . Differential gene expression in cultured retinoblastoma cells induced by cerulenin , a chemical inhibitor of P49327 , was evaluated by cDNA microarray analysis . Cerulenin treatment resulted in significant upregulation of cytochrome c ( P99999 ) by 1.2-fold , whereas S-phase kinase-associated protein-2 ( Q13309 ) , a negative regulator of cell cycle , and the lipid metabolic genes ( Q07869 , P19793 , and O00763 ) were significantly downregulated by -1.59- , -1.8- , -1.83- , and -1.5-fold , respectively , in comparison with untreated cancer cells . The expressions of key differentially expressed genes were confirmed by quantitative real-time PCR . The altered expression of genes involved in cell proliferation , cell signaling , apoptosis , and cell cycle , correlated with the anticancer effects of cerulenin . P49327 inhibition may thus be a potential strategy in retinoblastoma management . Puerarin decreases serum total cholesterol and enhances thoracic aorta endothelial nitric oxide synthase expression in diet-induced hypercholesterolemic rats . Hypercholesterolemia is a dominant risk factor for the development and progression of atherosclerosis and cardiovascular diseases . Natural compounds have been proved to be useful in lowering serum cholesterol to slow down the progression of cardiovascular diseases . Pueraria lobata is employed clinically to treat cardiovascular diseases in China . In the present study , the atheroscleroprotective potential of the herb 's major active compound , puerarin , was investigated by monitoring serum lipid profile and major enzyme expressions on cholesterol homeostasis in Sprague-Dawley rats fed with control diet , hypercholesterolmic diet or hypercholesterolmic diet plus administration of puerarin ( 300 mg/kg/day , p.o. ) for 4 weeks . Puerarin markedly attenuated the increased total cholesterol induced by hypercholesterolmic diet in both serum and liver . It caused a significant reduction in the atherogenic index . Expression of mRNA for hepatic 7alpha-hydroxylase ( P22680 ) was significantly enhanced but not for those of 3-hydroxy-3-methylglutaryl- DB01992 reductase ( HMGR ) and lanosterol 14alpha-demethylase ( Q16850 ) . To further explore the atheroscleroprotective potential of puerarin , acetylcholine induced endothelium-dependent vasorelaxation and endothelial nitric oxide synthase ( P29474 ) expression on isolated thoracic aortas were analyzed . Animals administered with puerarin suppressed the hypercholesterolemic diet induced impairment of P29474 expression , whereas there was no significant difference in the endothelium-dependent vasorelaxation among various groups of animals . These data indicated that puerarin reduced the atherogenic properties of dietary cholesterol in rats . Its hypocholesterolemic function may be due to the promotion of cholesterol and bile acids excretion in liver . Whether puerarin targets directly on cholesterol homeostasis or both cholesterol homeostasis and endothelial function remains to be determined . 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 . 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 . 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 . TGFbeta1 , TNFalpha , and insulin signaling crosstalk in regulation of the rat cholesterol 7alpha-hydroxylase gene expression . The TGFbeta1/Smad pathway plays a critical role in cholestasis and liver fibrosis . Previous studies show that TGFbeta1 , TNFalpha , and insulin inhibit cholesterol 7alpha-hydroxylase ( P22680 ) gene transcription and bile acid synthesis in human hepatocytes . In this study , we investigated insulin , TGFbeta1 , and TNFalpha regulation of rat Cyp7a1 gene transcription . In contrast to inhibition of human P22680 gene transcription , TGFbeta1 stimulates rat Cyp7a1 reporter activity . P84022 , FoxO1 , and HNF4alpha synergistically stimulated rat Cyp7a1 gene transcription . Mutations of the P84022 , FoxO1 , or HNF4alpha binding site attenuated the rat Cyp7a1 promoter activity . Furthermore , TNFalpha and cJun attenuated TGFbeta1 stimulation of rat Cyp7a1 . P01308 or adenovirus-mediated expression of constitutively active P31749 inhibited FoxO1 and P84022 synergy . In streptozotocin-induced diabetic rats , Cyp7a1 mRNA expression levels were induced and insulin attenuated P22680 mRNA levels . Chromatin immunoprecipitation assay showed that FoxO1 binding to Cyp7a1 chromatin was increased in diabetic rat livers and insulin reduced FoxO1 binding . These results suggest a mechanistic basis for induction of Cyp7a1 activity and bile acid synthesis in cholestatic rats and in diabetic rats . The crosstalk of insulin , TGFbeta and TNFalpha signaling pathways may regulate bile acid synthesis and lipid homeostasis in diabetes , fatty liver disease , and liver fibrosis . 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 . 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 . 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 . 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 . 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 . TFMPP may produce its stimulus effects via a P28222 mechanism . Tests of stimulus generalization were conducted using rats trained to discriminate 1.0 mg/kg of 1-(3-trifluoromethylphenyl)piperazine ( TFMPP ) from saline in a standard two-lever operant procedure . Generalization of the TFMPP-stimulus was found to occur with fenfluramine and 1-(3-chlorophenyl)-piperazine ( mCPP ) ; generalization did not occur with 8-OH DPAT , quipazine , LSD , 5-OMe Q08495 or 2,5- P28067 . Furthermore , the TFMPP-stimulus was not antagonized by pretreatment of the animals with tetrahydrotrazodone ( THT ) . Based on the results of these studies , and on the results of previous binding studies with these same agents , it is suggested that the stimulus properties of TFMPP are mediated primarily via a P28222 mechanism . 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 . 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 . [ 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 . 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 . 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 . [ 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 . 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 % . 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 . Impaired adipogenesis and lipolysis in the mouse upon selective ablation of the retinoid X receptor alpha mediated by a tamoxifen-inducible chimeric Cre recombinase ( Cre- P27361 ) in adipocytes . P19793 ( RXRalpha ) is involved in multiple signaling pathways , as a heterodimeric partner of several nuclear receptors . To investigate its function in energy homeostasis , we have selectively ablated the RXRalpha gene in adipocytes of 4-week-old transgenic mice by using the tamoxifen-inducible Cre- P27361 recombination system . Mice lacking RXRalpha in adipocytes were resistant to dietary and chemically induced obesity and impaired in fasting-induced lipolysis . Our results also indicate that RXRalpha is involved in adipocyte differentiation . Thus , our data demonstrate the feasibility of adipocyte-selective temporally controlled gene engineering and reveal a central role of RXRalpha in adipogenesis , probably as a heterodimeric partner for peroxisome proliferator-activated receptor gamma . Hypolipidemic effects of a new piperine derivative GB-N from Piper longum in high-fat diet-fed rats . CONTEXT : Long pepper , Piper longum Linn . ( Piperaceae ) , is widely used in traditional Mongolian medicine for treating hyperlipidemia and coronary heart disease . OBJECTIVE : To investigate the hypolipidemic effects of a new piperine derivative GB-N isolated from long pepper in high-fat diet-fed rats . METHODS : The levels of serum total cholesterol , triacylglycerols ( TG ) , low-density lipoprotein cholesterol ( LDL-C ) and high-density lipoprotein cholesterol ( HDL-C ) were determined by enzymatic colorimetric method . The levels of 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) , P22680 , lecithin cholesterol acyltransferase ( P04180 ) and P01130 ( P01130 ) mRNA and protein expression were detected by real-time polymerase chain reaction and western blot analysis . RESULTS AND DISCUSSION : Compared with model rats , oral administration of GB-N at doses of 2.5-10 mg/kg to hyperlipidemic rats could significantly decrease the levels of serum TG from 1.54 mmol/L in hyperlipidemic rats to 0.94-1.02 mmol/L , with an increase in serum HDL-C levels from 0.40 mmol/L in hyperlipidemic rats to 1.21-2.26 mmol/L . Treatment with GB-N ( 10 mg/kg ) could also significantly upregulate levels of hepatic P04035 , P22680 , P04180 and P01130 mRNA and protein expression . CONCLUSION : GB-N had hypolipidemic activity via regulating lipid metabolism pathways in liver of hyperlipidemic rats and could be explored as a potential agent for the prevention of hyperlipidemia diseases . 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 . 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 . 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 . No association between polymorphisms in four serotonin receptor genes , serotonin transporter gene and alcohol-related suicide . BACKGROUND : Serotonin ( 5-HT ) is an important neurotransmitter with wide-ranging functions . Its disfunction in the central nervous system seems to play an important role in many psychiatric disorders and suicidal behavior . The objective of this study was to examine the association between polymorphisms in different serotonin receptor genes ( HTR ) : P08908 ( polymorphism -1019C > G ) , P28222 ( polymorphisms 861G > C and -161A > T ) , P30939 ( polymorphism -78C > T ) and P28223 ( polymorphism -1420C > T ) , and serotonin transporter gene ( 5-HTT ) ( polymorphism LPR in promoter and VNTR in the second intron ) , and completed alcohol-related suicide , as well as between alcohol-dependent suicide victims . SUBJECTS AND METHODS : The study subjects were 373 Slovenian suicide victims ( mean age ± SD : 48.8 ± 17.7 years ) autopsied in the years 2002 through 2005 . During autopsy venous blood was drawn , and afterwards DNA extraction and alcoholimetric analysis were performed . Relatives of 79 suicide victims were interviewed using a semi-structured questionnaire designed according to Slovenian cultural and economic conditions . They provided information about the alcohol abuse of the suicide victims . Amongst the suicide victims were 25 alcohol misusers and 54 non-misusers . RESULTS : Association between polymorphisms in the selected serotonin receptor genes , transporter gene and completed alcohol-related suicide , as well as between alcohol-dependent suicide victims was not established . CONCLUSIONS : Present results suggest that selected polymorphisms of the 5-HT receptor genes and transporter gene are not involved in genetic susceptibility to completed suicide under acute influence of alcohol or among alcohol-dependent individuals , but further studies in a larger sample are needed . [ 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 . 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 . 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 . Fibroblast growth factor-2 in hyperplastic pituitaries of D2R knockout female mice . P14416 ( D2R ) knockout ( KO ) female mice develop chronic hyperprolactinemia and pituitary hyperplasia . Our objective was to study the expression of the mitogen fibroblast growth factor ( P09038 ) and its receptor , P11362 , comparatively in pituitaries from KO and wild-type ( WT ) female mice . We also evaluated P09038 subcellular localization and P09038 effects on pituitary function . P09038 -induced prolactin release showed a similar response pattern in both genotypes , even though basal and P09038 -stimulated release was higher in KO . P09038 stimulated pituitary cellular proliferation ( MTS assay and [(3)H]thymidine incorporation ) , with no differences between genotypes . P09038 concentration ( measured by ELISA ) in whole pituitaries or cultured cells was lower in KO ( P < 0.00001 and 0.00014 ) . Immunofluorescence histochemistry showed less P09038 in pituitaries from KO females and revealed a distinct P09038 localization pattern between genotypes , being predominantly nuclear in KO and cytosolic in WT pituitaries . Finally , P09038 could not be detected in the conditioned media from pituitary cultures of both genotypes . P11362 levels ( Western blot and immunohistochemistry ) were higher in pituitaries of KO . Basal concentration of phosphorylated ERKs was lower in KO cells ( P = 0.018 ) . However , when stimulated with P09038 , a significantly higher increment of P29323 phosphorylation was evidenced in KO cells ( P < or = 0.02 ) . We conclude that disruption of the D2R caused an overall decrease in pituitary P09038 levels , with an increased distribution in the nucleus , and increased P11362 levels . These results are important in the search for reliable prognostic indicators for patients with pituitary dopamine-resistant prolactinomas , which will make tumor-specific therapy possible . 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 . Establishment by adriamycin exposure of multidrug-resistant rat ascites hepatoma AH130 cells showing low P15559 activity and high cross resistance to mitomycins . A resistant subline ( AH130/5A ) selected from rat hepatoma AH130 cells after exposure to adriamycin ( P35318 ) showed remarkable resistance to multiple antitumor drugs , including mitomycin C ( DB00305 ) and porfiromycin ( PFM ) . PFM , vinblastine ( DB00570 ) , and P35318 accumulated in AH130/5A far less than in the parent AH130 ( AH130/P ) cells . AH130/5A cells showed overexpression of P-glycoprotein ( A6NDG6 ) , an increase in glutathione S-transferase activity , and a decrease in P15559 and glutathione peroxidase activity . The resistance to DB00305 and DB00570 of AH130/5A cells was partly reversed by H-87 , an inhibitor of A6NDG6 . Buthionine sulfoximine , an inhibitor of glutathione synthase , did not affect the action of DB00305 . tert-Butylhydroquinone induced P15559 activity , increased PFM uptake , and enhanced the growth-inhibitory action of DB00305 in AH130/5A cells . DB00266 , an inhibitor of P15559 , decreased PFM uptake and reduced the growth-inhibitory action of DB00305 in AH130/P cells . These results indicated that the adriamycin treatment of hepatoma cells caused multifactorial multidrug resistance involving a decrease in P15559 activity . 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 . P01308 activates the alpha isoform of class II phosphoinositide 3-kinase . The novel class II phosphoinositide ( PI ) 3-kinases are characterized by the presence of a C-terminal P06681 domain , but little is known about their regulation . We find insulin causes a rapid 2-3-fold increase in the activity of PI 3-kinase C2alpha ( PI3K-C2alpha ) in CHO-IR cells , 3T3- Q9NUQ9 adipocytes , and fully differentiated L5L6 myotubes . No insulin-induced activation of PI3K-C2alpha was observed in cell types known to have low responsiveness to insulin including P29320 293 cells , 3T3- Q9NUQ9 preadipocytes , and undifferentiated L5L6 myoblasts . The mechanism of activation of PI3K-C2alpha by insulin differs from that of class Ia PI 3-kinases in that insulin stimulation did not cause PI3K-C2alpha to associate with P35568 or insulin receptor . PI3K-C2alpha existed as a doublet , and insulin stimulation caused a redistribution from the lower molecular weight band to the higher molecular weight band , suggesting phosphorylation-induced bandshift . Consistent with this , in vitro phosphatase treatment reduced the intensity of the upper band back to that seen in unstimulated cells . This suggests that insulin-induced phosphorylation could play a role in regulation of the activity of PI3K-C2alpha . The finding that insulin activates PI3K-C2alpha in cell types known to possess a wide range of responses to insulin suggests that PI3K-C2alpha is a novel component of insulin-stimulated signaling cascades . 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 . DB06155 , a cannabinoid receptor type 1 inverse agonist , inhibits hepatocyte lipogenesis by activating liver kinase B1 and AMP-activated protein kinase axis downstream of Gα i/o inhibition . Liver X receptor-α ( LXRα ) and its target sterol regulatory element-binding protein-1c ( SREBP-1c ) play key roles in hepatic lipogenesis . DB06155 , an inverse agonist of cannabinoid receptor type 1 ( P21554 ) , has been studied as an antiobesity drug . In view of the link between P21554 and energy metabolism , this study investigated the effect of rimonabant on LXRα-mediated lipogenesis in hepatocytes and the underlying basis . DB06155 treatment inhibited P22680 -LXRα response element gene transactivation and an increase in LXRα mRNA level by the LXRα agonist N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide ( T0901317 ) in HepG2 cells . DB06155 consistently attenuated the activation of SREBP-1c and its target gene induction . The reversal by P21554 agonists on rimonabant 's repression of SREBP-1c supported the role of P21554 in this effect . DB06155 inhibited the activation of SREBP-1c presumably via Gα(i/o) inhibition , as did pertussis toxin . Adenylyl cyclase activator forskolin or 8-bromo- DB02527 treatment mimicked the action of rimonabant , suggesting that Gα(i/o) inhibition causes repression of SREBP-1c by increasing the DB02527 level . Knockdown or chemical inhibition of protein kinase A ( PKA ) prevented the inhibition of LXRα by rimonabant , supporting the fact that an increase in DB02527 content and PKA activation , which catalyzes LXRα inhibitory phosphorylation , might be responsible for the antilipogenic effect . In addition , rimonabant activated liver kinase B1 ( Q15831 ) , resulting in the activation of AMP-activated protein kinase responsible for LXRα repression . Moreover , PKA inhibition prevented the activation of Q15831 , supporting the fact that PKA regulates Q15831 . In conclusion , rimonabant has an antilipogenic effect in hepatocytes by inhibiting LXRα-dependent SREBP-1c induction , as mediated by an increase in PKA activity and PKA-mediated Q15831 activation downstream of P21554 -coupled Gα(i/o) inhibition . 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 .

MH_train_1111

MH_train_1111

MH_train_1111

interacts_with DB00163?

multiple_choice

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 . 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 . 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 . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . P28300 -like-2 promotes tumour angiogenesis and is a potential therapeutic target in angiogenic tumours . P28300 -like 2 ( Q9Y4K0 ) , a secreted enzyme that catalyzes the cross-linking of collagen , plays an essential role in developmental angiogenesis . We found that administration of the Q9Y4K0 -neutralizing antibody AB0023 inhibited P09038 -induced angiogenesis in Matrigel plug assays and suppressed recruitment of angiogenesis promoting bone marrow cells . Small hairpin RNA-mediated inhibition of Q9Y4K0 expression or inhibition of Q9Y4K0 using AB0023 reduced the migration and network-forming ability of endothelial cells , suggesting that the inhibition of angiogenesis results from a direct effect on endothelial cells . To examine the effects of AB0023 on tumour angiogenesis , AB0023 was administered to mice bearing tumours derived from SKOV-3 ovarian carcinoma or Lewis lung carcinoma ( LLC ) cells . AB0023 treatment significantly reduced the microvascular density in these tumours but did not inhibit tumour growth . However , treatment of mice bearing SKOV-3-derived tumours with AB0023 also promoted increased coverage of tumour vessels with pericytes and reduced tumour hypoxia , providing evidence that anti- Q9Y4K0 therapy results in the normalization of tumour blood vessels . In agreement with these data , treatment of mice bearing LLC-derived tumours with AB0023 improved the perfusion of the tumour-associated vessels as determined by ultrasonography . Improved perfusion and normalization of tumour vessels after treatment with anti-angiogenic agents were previously found to improve the delivery of chemotherapeutic agents into tumours and to result in an enhancement of chemotherapeutic efficiency . Indeed , treatment with AB0023 significantly enhanced the anti-tumourigenic effects of taxol . Our results suggest that inhibition of Q9Y4K0 may prove beneficial for the treatment of angiogenic tumours . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Protein kinase C-beta II Is an apoptotic lamin kinase in polyomavirus-transformed , etoposide-treated pyF111 rat fibroblasts . The role of protein kinase C-beta(II) ( P05771 (II) ) in etoposide ( DB00773 ) -induced apoptosis was studied using polyomavirus-transformed pyF111 rat fibroblasts in which P05771 (II) specific activity in the nuclear membrane ( NM ) doubled and the enzyme was cleaved into catalytic fragments . No P05771 (II) complexes with lamin B1 and/or active caspases were immunoprecipitable from the NM of proliferating untreated cells , but large complexes of P05771 (II) holoprotein and its catalytic fragments with lamin B1 , active caspase-3 and -6 , and inactive phospho-CDK-1 , but not P05771 (I) or PKC-delta , could be immunoprecipitated from the NM of DB00773 -treated cells , suggesting that P05771 (II) is an apoptotic lamin kinase . By 30 min after normal nuclei were mixed with cytoplasms from DB00773 -treated , but not untreated , cells , P05771 (II) holoprotein had moved from the apoptotic cytoplasm to the normal NM , and lamin B1 was phosphorylated before cleavage by caspase-6 . Lamin B1 phosphorylation was partly reduced , but its cleavage was completely prevented , despite the presence of active caspase-6 , by adding a selective PKC-betas inhibitor , hispidin , to the apoptotic cytoplasms . Thus , a P05771 (II) response to DB00773 seems necessary for lamin B1 cleavage by caspase-6 and nuclear lamina dissolution in apoptosing pyF111 fibroblasts . The possibility of P05771 (II) being an apoptotic lamin kinase in these cells was further suggested by lamin B1-bound PKC-delta being inactive or only slightly active and by P17252 not combining with the lamin . Alpha-tocopherol prevents cyclosporin A-mediated activation of phospholipase A2 and inhibition of Na+ , K(+)-adenosine triphosphatase activity in cultured hamster renal tubular cells . At concentrations of 0.5 microM and upward , cyclosporin A ( DB00091 ) caused dose-related inhibition of the growth of a hamster renal tubular cell line ( Q86TB3 ATCC ; Q16663 ) in vitro . Inhibition of cell growth was due to the cytotoxic properties of DB00091 which were associated with enhancement of activity of phospholipase A2 ( P04054 ) according to the increased generation of arachidonic acid and lysophosphatidylcholine ( Q16549 ) . Arachidonate per se , at concentrations of up to 20 microM , did not affect the growth of Q86TB3 cells , while cyclooxygenase and P09917 inhibitors failed to protect the cells against the antiproliferative effects of DB00091 . However , Q16549 caused dose-related inhibition of the growth of Q86TB3 cells . Moreover , coincubation with lysophospholipase or DB00163 ( AT , vitamin E ) , a P04054 inhibitory and lysophospholipid-complexing agent , protected the Q86TB3 cells against both DB00091 and Q16549 . The Na+ , K(+)-ATPase activity of Q86TB3 cells was also inhibited by DB00091 , with the enzyme being protected by inclusion of AT or lysophospholipase . Increased activity of P04054 and inhibition of Na+ , K(+)-ATPase preceded cytotoxicity and cytolysis . Excessive production of lysophospholipids and consequent inhibition of Na+ , K(+)-ATPase in renal tubular cells is a possible mechanism of DB00091 -induced nephrotoxicity . The protective effects of AT suggest that this agent may be clinically useful in preventing the renal side effects of DB00091 . P28300 plays a critical role in endothelial cell stimulation to drive tumor angiogenesis . Identification of key molecules that drive angiogenesis is critical for the development of new modalities for the prevention of solid tumor progression . Using multiple models of colorectal cancer , we show that activity of the extracellular matrix-modifying enzyme lysyl oxidase ( P28300 ) is essential for stimulating endothelial cells in vitro and angiogenesis in vivo . We show that P28300 activates Akt through platelet-derived growth factor receptor β ( PDGFRβ ) stimulation , resulting in increased P15692 expression . P28300 -driven angiogenesis can be abrogated through targeting P28300 directly or using inhibitors of PDGFRβ , Akt , and P15692 signaling . Furthermore , we show that P28300 is clinically correlated with P15692 expression and blood vessel formation in 515 colorectal cancer patient samples . Finally , we validate our findings in a breast cancer model , showing the universality of these observations . Taken together , our findings have broad clinical and therapeutic implications for a wide variety of solid tumor types . 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 . 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 . 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 ) . Genetics of type 2 diabetes mellitus and other specific types of diabetes ; its role in treatment modalities . Type 2 diabetes mellitus ( T2DM ) is among the most challenging health issues of the 21st century and is associated with an alarming rise in the incidence . The pathophysiological processes that lead to development of T2DM are still unclear , however impairment in insulin secretion and/or action is clearly indicated . Type 2 diabetes is a polygenic disorder with multiple genes located on different chromosomes contributing to its susceptibility . Analysis of the genetic factors is further complicated by the fact that numerous environmental factors interact with genes to produce the disorder . Only a minority of cases of type 2 diabetes are caused by single gene defects and one example is maturity onset diabetes of the young ( MODY ) . Previous studies indicated that variants in genes encoding the pancreatic β-cell K+ DB00171 channel subunits Kir6.2 ( Q14654 ) and Q09428 ( Q09428 ) are associated with neonatal diabetes . Six different types of maturity onset diabetes of young ( MODY ) have been identified based on characteristic gene defect . The common Pro12Ala polymorphism in peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) gene was confirmed in several studies to be associated with type 2 diabetes as well . More recently , studies reported variants within a novel gene , Q9NQB0 , as a putative susceptibility gene for type 2 diabetes across many ethnic backgrounds around the world . MODY patients respond better to sulphonylureas and metformin , while neonatal diabetes patients with genetic mutations can be changed from insulin to oral drugs . We hereby provide a comprehensive review on the role of genetics in type 2 diabetes mellitus . Expression of ras proto-oncogenes in the Dunning R3327 rat prostatic adenocarcinoma system . Steady-state levels of c-Ha-ras mRNA were measured in eight sublines of the Dunning R3327 rat prostatic adenocarcinoma . As a control , normal dorsal prostate tissue was studied . Increased expression of c-Ha-ras is associated with tumor progression in one lineage of the Dunning R3327 system ( H to AT1 to P24752 -Lu and P24752 -Ly-Lu ) . Here ras mRNA increases as the tumor advances from androgen dependence and a high degree of differentiation to an anaplastic aneuploid phenotype with high metastatic potential . However , in the other Dunning lineage ( H to HI to HI-F to P01008 ) , expression of c-Ha-ras is variable and does not correlate with tumor progression . Immunocytochemistry showed that levels of the c-Ha-ras P38936 protein paralleled steady-state mRNA levels in variants . Transfection assays , using NIH/3T3 cells , suggested that the ras loci were not activated in the R3327 tumors . Levels of P01116 mRNA were also measured in the Dunning tumors ; these did not correlate with tumor progression in either lineage . Expression of N-ras mRNA was not detected in the Dunning tumors . 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 . DB00163 , P09917 and oxidative stress in haemodialysis patients : facts , not fancies . 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 . Fibroblast growth factors mobilize peritoneal macrophage intracellular calcium . Macrophages have been implicated in the propagation of inflammatory disease . The evidence linking macrophages to inflammation stems from their elicited responses to various extracellular ligands eventually culminating in the elaboration of a variety of inflammatory mediators . As part of an investigation of fibroblast growth factors role in promoting inflammation , we examined one aspect of transmembrane signal transduction , intracellular calcium mobilization following culture of murine peritoneal macrophages with acidic and basic fibroblast growth factor . Peritoneal macrophages displayed a rapid rise in cytosolic calcium from a basal level of 147.6 +/- 25.4 nM to 261.9 +/- 49.9 nM at 3.5 minutes following culture with acidic fibroblast growth . A similar rise in calcium was noted with basic fibroblast growth factor . Titration revealed the maximal effective dose of P05230 and P09038 with respect to calcium response to be 10 ng/ml . Using blockers of both voltage and non-voltage gated channels , the FGF induced rise in cytosolic calcium was specifically abolished . Similarly , using specific P09917 ( A69412 ) or cyclooxygenase ( Indomethacin ) blockers , the P05230 induced rise in maximal calcium response was reduced by 41 % and 96 % respectively . On the basis of these data , we speculate on some possible roles that FGF may play in the inflammatory response . Growth rates of hprt and tk mutant CHO cell lines . The growth rates of 31 X-ray-induced hypoxanthine phosphoribosyl transferase ( hprt ) deficient mutants of CHO- P04264 cells were measured . Mutants had been classified as ( 1 ) full-deletion , ( 2 ) partial deletion or rearrangement , or ( 3 ) unchanged by Southern blot analyses . No relationship between growth rate and deletion type was observed . Even where all hprt-specific bands were missing , proliferation rates in culture were normal . Additionally , in CHO- P01008 -2 cells , which are heterozygous at the tk locus , no difference in growth rates between a spontaneous hprt mutant and its parent was observed , although double hprt-tk-/- mutants grew more slowly . DB00163 80th anniversary : a double life , not only fighting radicals . Recent research on DB00163 has revealed specific cellular functions of this compound belonging to the vitamin E family . Alpha-tocopherol can act as a radical scavenger , as a pro-oxidant , as an anti-alkylation agent and , most important , by mechanisms that are independent of the above properties . To the last group belong protein kinase C and P09917 inhibition at post-translational level , as well as DB00163 activation of protein phosphatase 2A and diacylglycerol kinase . Furthermore , at transcriptional level , several genes ( P16671 , alpha-TTP , alpha-tropomyosin , and collagenase ) are modulated by DB00163 . These effects result in inhibition of smooth muscle cell proliferation , platelet aggregation , and monocyte adhesion and may be related to the alleged protection of atherosclerosis by vitamin E. On the other side , epidemiological and intervention studies have shown some inconsistent results . Rather than disregarding vitamin E as a means to protect against atherosclerosis progression , it would be wiser to better design clinical trials based on current knowledge of the biological properties of the molecule . 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 . 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 . [ Effect of tocopherol and tocopherol quinone complexes with proteins on activity of leukotriene B4 lipoxygenase ] . While estimating the effect of tocopherol , tocopherylquinone and their complexes with the tocopherol-binding proteins from the rat liver cytosole on arachidonate P09917 from peritoneal-lymphocytes and soybean linoleate- P09917 DB00163 and especially its complex with tocopherol-binding protein was defined to inhibit the activity of both vegetative- and animal nature-lipoxygenase . 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 . Overexpression of cytochrome P450 4F2 in mice increases 20-hydroxyeicosatetraenoic acid production and arterial blood pressure . P78329 ( P78329 ) activity is thought to be a factor in the pathogenesis of hypertension through its bioactive metabolite 20-hydroxyeicosatetraenoic acid ( 20-HETE ) . We previously found that a gain-in-function P78329 variant in a Chinese cohort was associated with elevated urinary 20-HETE and hypertension . To further explore this association we generated a transgenic mouse model expressing P78329 driven by a modified mouse kidney androgen-regulated protein promoter . This heterologous promoter regulated the expression of luciferase and his-tagged P78329 in transfected P29320 293 cells . In the kidney of transgenic mice , P78329 was localized to renal proximal tubule epithelia and was expressed at a higher level than in control mice , leading to increased urinary 20-HETE excretion . Assessment of P78329 activity by an arachidonic acid hydroxylation assay showed that 20-HETE production was significantly higher in kidney microsomes of transgenic mice compared to control mice , as was their systolic blood pressure . There was a positive correlation of blood pressure with urinary 20-HETE levels . Our results show that increased expression of P78329 in mice enhanced 20-HETE production and elevated blood pressure . Effects of long-term elevated glucose on collagen formation by mesangial cells . Glomerulosclerosis is one of the complications of diabetes that occurs after many years of uncontrolled hyperglycemia . Mesangial cells ( MCs ) exposed to high glucose ( HG ) for short periods have shown that transforming growth factor-beta ( TGF-beta ) and activated diacylglycerol-dependent protein kinase C ( PKC ) mediate increased collagen formation . Our study examined collagen formation by MCs exposed to HG for 8 weeks . Exposure to HG in overnight culture resulted in the activation of all PKC isoforms . In contrast , 8-week exposure to HG resulted in the persistent activation of PKC-delta , did not change P17252 or -beta activity , and decreased PKC-epsilon activity while increasing collagen I and IV gene and protein expression . Collagen IV accumulation was reversed by specific PKC-delta inhibition . Collagen IV gene expression was completely normalized by TGF-beta neutralization ; however , this was associated with plasminogen activator inhibitor-1 ( P05121 ) overexpression and a modest reduction in collagen protein . Our studies suggest that prolonged exposure to HG results in PKC-delta-driven collagen accumulation by MCs mediated by P05121 but independent of TGF-beta . 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 . 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 . Anti-atherogenic and anti-angiogenic activities of polyphenols from propolis . Propolis is a polyphenol-rich resinous substance extensively used to improve health and prevent diseases . The effects of polyphenols from different sources of propolis on atherosclerotic lesions and inflammatory and angiogenic factors were investigated in P01130 gene ( LDLr-/- ) knockout mice . The animals received a cholesterol-enriched diet to induce the initial atherosclerotic lesions ( IALs ) or advanced atherosclerotic lesions ( AALs ) . The IAL or AAL animals were divided into three groups , each receiving polyphenols from either the green , red or brown propolis ( 250 mg/kg per day ) by gavage . After 4 weeks of polyphenol treatment , the animals were sacrificed and their blood was collected for lipid profile analysis . The atheromatous lesions at the aortic root were also analyzed for gene expression of inflammatory and angiogenic factors by quantitative real-time polymerase chain reaction and immunohistochemistry . All three polyphenol extracts improved the lipid profile and decreased the atherosclerotic lesion area in IAL animals . However , only polyphenols from the red propolis induced favorable changes in the lipid profiles and reduced the lesion areas in AAL mice . In IAL groups , VCAM , P13500 , FGF , PDGF , P15692 , PECAM and P14780 gene expression was down-regulated , while the metalloproteinase inhibitor P01033 gene was up-regulated by all polyphenol extracts . In contrast , for advanced lesions , only the polyphenols from red propolis induced the down-regulation of P16671 and the up-regulation of P09601 and P01033 when compared to polyphenols from the other two types of propolis . In conclusion , polyphenols from propolis , particularly red propolis , are able to reduce atherosclerotic lesions through mechanisms including the modulation of inflammatory and angiogenic factors . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 ) Antagonism by salvianolic acid B of lipopolysaccharide-induced disseminated intravascular coagulation in rabbits . The aim of the present study was to investigate the effects of salvianolic acid B on lipopolysaccharide ( LPS ) -induced disseminated intravascular coagulation ( DIC ) in rabbits . Continuous infusion of LPS was used to induce a DIC model in rabbits . Treatment with salvianolic acid B ( 1 , 3 or 6 mg/kg ) was started simultaneously with LPS infusion ( 0.5 mg/kg LPS in 60 mL saline ; 10 mL/h over a period of 6 h ) through the contralateral marginal ear vein . Activated partial thromboplastin time ( APTT ) , prothrombin time ( PT ) , platelet count and fibrinogen concentration were determined , as were plasma levels of fibrin-fibrinogen degradation products ( Q9NRC9 ) , alanine aminotransferase ( ALT ) , blood urea nitrogen ( BUN ) , protein C activity , antithrombin III ( P01008 ) and tumour necrosis factor ( P01375 ) -α concentration . The gradual impairment of haemostatic parameters was induced by continuous infusion of LPS . There were marked increases in APTT , PT , BUN , ALT and plasma P01375 -α and marked decreases in the platelet count , fibrinogen , Q9NRC9 , protein C and P01008 . The intravenous administration of 1 , 3 or 6 mg/kg salvianolic acid B attenuated the increases in APTT , PT , BUN , ALT and plasma P01375 -α and the decreases in fibrinogen , platelet , Q9NRC9 , protein C and P01008 induced by LPS infusion . These observations indicate that salvianolic acid B has an effect against LPS-induced DIC in rabbits . 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 . Inactivation of androgens by UDP-glucuronosyltransferases in the human prostate . In the human prostate , dihydrotestosterone ( DB02901 ) -- the natural androgen having the highest affinity for the androgen receptor -- is not released directly into the systemic circulation from peripheral target tissues but it is rather converted in situ into two metabolites which have a low affinity for the androgen receptor : androsterone ( ADT ) and androstane-3alpha,17beta-diol ( 3alpha-DIOL ) . Several clinical observations indicate that these two androgen metabolites are further inactivated in the prostate by glucuronidation . In the human , the family of UDP-glucuronosyltransferase ( P78381 ) enzymes comprises 18 members in three subfamilies : P22309 , UGT2A and UGT2B . Identification of the substrates for each member has revealed that three UGT2B enzymes are mainly responsible for DB02901 , ADT and 3alpha-DIOL glucuronidation : P16662 , P54855 and O75795 . Tissue distribution and cellular localization of UGT2B transcripts and proteins clearly indicate that only P54855 and O75795 are expressed in the prostate . Using the human prostate carcinoma LNCaP cell line , it was shown that UGT2B expression and activity are negatively regulated by several factors , including androgens . On the other hand , inhibition of UGT2B115/17 expression by small interfering RNA ( siRNA ) resulted in an induced response to DB02901 of androgen-receptor target genes such as PSA , Q9Y5K2 , Q99801 , O15393 , O15403 and P15692 . It is suggested that the conjugating activity of P78381 enzymes in androgen target tissues is a mechanism for modulating the action of steroids and/or protecting the tissues from deleterious high concentrations of androgens . Genetic determinants influencing human serum metabolome among African Americans . Phenotypes proximal to gene action generally reflect larger genetic effect sizes than those that are distant . The human metabolome , a result of multiple cellular and biological processes , are functional intermediate phenotypes proximal to gene action . Here , we present a genome-wide association study of 308 untargeted metabolite levels among African Americans from the Atherosclerosis Risk in Communities ( ARIC ) Study . Nineteen significant common variant-metabolite associations were identified , including 13 novel loci ( p < 1.6 × 10(-10) ) . These loci were associated with 7-50 % of the difference in metabolite levels per allele , and the variance explained ranged from 4 % to 20 % . Fourteen genes were identified within the nineteen loci , and four of them contained non-synonymous substitutions in four enzyme-encoding genes ( P03952 , Q9HAT2 , P31327 , and Q9UHE5 ) ; the other significant loci consist of eight other enzyme-encoding genes ( P12821 , P50440 , Q96HD9 , Q68CK6 , Q5T1C6 , P08319 , P22309 , O43280 ) , a transporter gene ( Q9NSD5 ) and a polycystin protein gene ( Q9P0L9 ) . In addition , four potential disease-associated paths were identified , including two direct longitudinal predictive relationships : Q9UHE5 with N-acetylornithine , N-acetyl-1-methylhistidine and incident chronic kidney disease , and O43280 with trehalose and incident diabetes . These results highlight the value of using endophenotypes proximal to gene function to discover new insights into biology and disease pathology . 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 . 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 . 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 . 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 . 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 . New gene variants associated with venous thrombosis : a replication study in White and Black Americans . BACKGROUND : We evaluated 10 single-nucleotide polymorphisms ( SNPs ) identified in three European case-control studies as risk factors for venous thrombosis . OBJECTIVES : We sought to replicate the positive findings from this report among Whites and to evaluate the association of these SNPs with venous thrombosis for the first time among Blacks . PATIENT/METHODS : These SNPs were evaluated in a case-control study of deep vein thrombosis and pulmonary embolism that included 1076 cases and 1239 controls . About 50 % of subjects were African Americans . We measured plasma factor ( F ) XI on a subset of subjects . RESULTS : Among Whites , positive findings for rs13146272 in the Q6ZWL3 gene , for rs3087505 in the P03952 gene and for rs3756008 and rs2036914 in the P03951 gene were found . We did not find significant associations for rs2227589 in the P01008 gene and for rs1613662 in the Q9HCN6 gene . Among Blacks , rs2036914 in P03951 and rs670659 in P49802 were related to venous thrombosis , but the study had limited statistical power for many SNPs . Among Blacks , plasma P03951 was related to two SNPs and the OR relating to the 90th percentile of the control distribution of plasma P03951 was 2.6 ( 95 % CI , 1.4 , 5.0 ) . CONCLUSIONS : Our study supports the finding that genetic variants in the P03951 gene are risk factors for venous thrombosis among both Whites and Blacks , although the findings in Blacks require confirmation . A meta-analysis of five case-control studies indicates that rs2227589 in the P01008 gene , rs13146272 in the Q6ZWL3 gene and rs1613662 in the Q9HCN6 gene are risk factors for venous thrombosis among Whites . 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 .

MH_train_1112

MH_train_1112

MH_train_1112

interacts_with DB06603?

multiple_choice

Effect of 27nt small RNA on endothelial nitric-oxide synthase expression . We have reported previously that the 27nt repeat polymorphism in endothelial nitric-oxide synthase ( P29474 ) intron 4 -- a source of 27nt small RNA -- inhibits P29474 expression . In the current study , we have investigated how 27nt small RNA suppresses P29474 expression . Using a chromatin immunoprecipitation assay , we examined histone acetylation in the 27nt repeat element of P29474 intron 4 , the promoter region up to -1486 bp , and the 5' enhancer region ( -4583/-4223bp ) in human aortic endothelial cells ( HAECs ) treated with 27nt RNA duplex . 27nt RNA duplex induced hyperacetylation in H3 ( lysine8 , 12 , and 23 ) and H4 ( lysine 9 and 12 ) at the 27nt repeat element , which then interacted with nuclear actin , histone deacetylase 3 ( O15379 ) , and NonO proteins . In contrast , the histone H3 and H4 became hypoacetylated at the P29474 core promoter . HAECs treated with 27nt RNA duplex had reduced P29474 expression , but treatment with either O15379 small interfering RNA or NonO siRNA significantly attenuated the 27nt small RNA-induced suppression . We further found that 27nt small RNA induced DNA methylation in a region approximately 750nt upstream of the intron 4 repeats , and a methyltransferase inhibitor reversed the effect on methylation and P29474 expression . Our study demonstrates that 27nt small RNA may suppress P29474 expression by altering histone acetylation and DNA methylation in regions adjacent to the 27nt repeat element and core promoter . 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 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 . 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 . 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 . 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 . 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 . 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 . c-Jun controls histone modifications , NF-kappaB recruitment , and RNA polymerase II function to activate the ccl2 gene . Interleukin-1 ( IL-1 ) -induced mRNA expression of ccl2 ( also called P13500 ) , a prototypic highly regulated inflammatory gene , is severely suppressed in cells lacking c-Jun or Jun N-terminal protein kinase 1 ( P45983 ) / P45984 genes and is only partially restored in cells expressing a c-Jun(SS63/73AA) mutant protein . We used chromatin immunoprecipitation to identify three c-Jun-binding sites located in the far 5' region close to the transcriptional start site and in the far 3' region of murine and human ccl2 genes . Mutational analysis revealed that the latter two sites contribute to ccl2 transcription in response to the presence of IL-1 or of ectopically expressed c-Jun- P39905 -2 dimers . Further experiments comparing wild-type and c-Jun-deficient cells revealed that c-Jun regulates Ser10 phosphorylation of histone H3 , acetylation of histones H3 and H4 , and recruitment of histone deacetylase 3 ( O15379 ) , NF-kappaB subunits , and RNA polymerase II across the ccl2 locus . c-Jun also coimmunoprecipitated with p65 NF-kappaB and O15379 . Based on DNA microarray analysis , c-Jun was required for full expression of 133 out of 162 IL-1-induced genes . For inflammatory genes , these data support the idea of an activator function of c-Jun that is executed by multiple mechanisms , including phosphorylation-dependent interaction with p65 NF-kappaB and O15379 at the level of chromatin . Class I histone deacetylase-mediated repression of the proximal promoter of the activity-regulated cytoskeleton-associated protein gene regulates its response to brain-derived neurotrophic factor . We examined the transcriptional regulation of the activity-regulated cytoskeleton-associated protein gene ( Arc ) , focusing on P23560 -induced Arc expression in cultured rat cortical cells . Although the synaptic activity-responsive element ( SARE ) , located -7 kbp upstream of the Arc transcription start site , responded to DB01221 , P23560 , or P09038 , the proximal region of the promoter ( Arc/-1679 ) was activated by P23560 or P09038 , but not by DB01221 , suggesting the presence of at least two distinct Arc promoter regions , distal and proximal , that respond to extracellular stimuli . Specificity protein 4 ( Q02446 ) and early growth response 1 ( P18146 ) controlled Arc/-1679 transcriptional activity via the region encompassing -169 to -37 of the Arc promoter . We found that trichostatin A ( P32119 ) , a histone deacetylase ( HDAC ) inhibitor , significantly enhanced the inductive effects of P23560 or P09038 , but not those of DB01221 on Arc expression . Inhibitors of class I/IIb HDACs , DB02546 , and class I HDACs , MS-275 , but not of class II HDACs , MC1568 , enhanced P23560 -induced Arc expression . The enhancing effect of P32119 was mediated by the region from -1027 to -1000 bp , to which serum response factor ( P11831 ) and Q13547 bound . The binding of Q13547 to this region was reduced by P32119 . Thus , Arc expression was suppressed by class I HDAC-mediated mechanisms via chromatin modification of the proximal promoter whereas the inhibition of HDAC allowed Arc expression to be markedly enhanced in response to P23560 or P09038 . These results contribute to our understanding of the physiological role of Arc expression in neuronal functions such as memory consolidation . 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 . 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 . 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 . 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 . Q13547 ( Q13547 ) participates in the down-regulation of corticotropin releasing hormone gene ( crh ) expression . The paraventricular nucleus of the hypothalamus ( PVH ) plays a central role in regulating the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . Medial parvocellular neurons of the PVH ( mpPVH ) integrate sensory and humoral inputs to maintain homeostasis . Humoral inputs include glucocorticoids secreted by the adrenals , which down-regulate Q9Y251 activation . A primary glucocorticoid target is the population of mpPVH neurons that synthesize and secrete corticotropin-releasing factors , the most potent of which is corticotropin-releasing hormone ( P06850 ) . Although P06850 gene ( crh ) expression is known to be down-regulated by glucocorticoids , the mechanisms by which this process occurs are still poorly understood . To begin this study we postulated that glucocorticoid repression of crh involves HDAC recruitment to the region of the crh proximal promoter . To evaluate this hypothesis , we treated hypothalamic cells that express P06850 with the HDAC inhibitor trichostatin A ( P32119 ) . As predicted , treatment with P32119 led to increased P06850 mRNA levels and crh promoter activity . Although co-treatment with DB00514 ( 10(-7)M ) reduced the P32119 effect on mRNA levels , it failed to reduce promoter activity ; however co-transfection of Q13547 but not 3 restored DB00514 inhibition . A distinction between Q13547 and 3 was also apparent with respect to crh promoter occupancy . DB00514 led to increased Q13547 but not O15379 occupancy . In vivo studies revealed that P06850 -immunoreactive ( -ir ) neurons contained Q13547 - and O15379 -ir . Collectively , these data point to a role for Q13547 in the physiologic regulation of crh . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Plasma membrane cholesterol content affects nitric oxide diffusion dynamics and signaling . DB00435 ( NO ) signaling is inextricably linked to both its physical and chemical properties . Due to its preferentially hydrophobic solubility , NO molecules tend to partition from the aqueous milieu into biological membranes . We hypothesized that plasma membrane ordering provided by cholesterol further couples the physics of NO diffusion with cellular signaling . Fluorescence lifetime quenching studies with pyrene liposome preparations showed that the presence of cholesterol decreased apparent diffusion coefficients of NO approximately 20-40 % , depending on the phospholipid composition . Electrochemical measurements indicated that the diffusion rate of NO across artificial bilayer membranes were inversely related to cholesterol content . Sterol transport-defective Niemann-Pick type C1 ( O15118 ) fibroblasts exhibited increased plasma membrane cholesterol content but decreased activation of both intracellular soluble guanylyl cyclase and vasodilator-stimulated phosphoprotein ( P50552 ) phosphorylation at DB00133 (239) induced by exogenous NO exposure relative to their normal human fibroblast ( NHF ) counterparts . Augmentation of plasma membrane cholesterol in NHF diminished production of both cGMP and P50552 phosphorylation elicited by NO to O15118 -comparable levels . Conversely , decreasing membrane cholesterol in O15118 resulted in the augmentation in both cGMP and P50552 phosphorylation to a level similar to those observed in NHF . Increasing plasma membrane cholesterol contents in NHF , platelets , erythrocytes and tumor cells also resulted in an increased level of extracellular diaminofluorescein nitrosation following NO exposure . These findings suggest that the impact of cholesterol on membrane fluidity and microdomain structure contributes to the spatial heterogeneity of NO diffusion and signaling . 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 . 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 . Imatinib triggers mesenchymal-like conversion of CML cells associated with increased aggressiveness . Chronic myelogenous leukemia ( CML ) is a cytogenetic disorder resulting from the expression of p210BCR- P00519 . Imatinib , an inhibitor of P11274 - P00519 , has emerged as the leading compound to treat CML patients . Despite encouraging clinical results , resistance to imatinib represents a major drawback for therapy , as a substantial proportion of patients are refractory to this treatment . Recent publications have described the existence of a small cancer cell population with the potential to exhibit the phenotypic switch responsible for chemoresistance . To investigate the existence of such a chemoresistant cellular subpopulation in CML , we used a two-step approach of pulse and continuous selection by imatinib in different CML cell lines that allowed the emergence of a subpopulation of adherent cells ( IM-R Adh ) displaying an epithelial-mesenchymal transition ( EMT ) -like phenotype . Overexpression of several EMT markers was observed in this CML subpopulation , as well as in P28906 (+) CML primary cells from patients who responded poorly to imatinib treatment . In response to imatinib , this P16070 (high)/ P25063 (low) IM-R Adh subpopulation exhibited increased adhesion , transmigration and invasion in vitro and in vivo through specific overexpression of the αVβ3 receptor . Q05397 /Akt pathway activation following integrin β3 ( ITGβ3 ) engagement mediated the migration and invasion of IM-R Adh cells , whereas persistent activation of P29323 counteracted P11274 - P00519 inhibition by imatinib , promoting cell adhesion-mediated resistance . 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 . Design of chimeric histone deacetylase- and tyrosine kinase-inhibitors : a series of imatinib hybrides as potent inhibitors of wild-type and mutant P11274 - P00519 , PDGF-Rbeta , and histone deacetylases . Inhibitors of histone deacetylases are a new class of cancer therapeutics with possibly broad applicability . Combinations of HDAC inhibitors with the kinase inhibitor 1 ( Imatinib ) in recent studies showed additive and synergistic effects . Here we present a new concept by combining inhibition of protein kinases and HDACs , two independent pharmacological activities , in one synthetic small molecule . In general , the HDAC inhibition profile , the potencies , and the probable binding modes to Q13547 and Q9UBN7 were similar as for 6 ( DB02546 ) . Inhibition of Abl kinase in biochemical assays was maintained for most compounds , but in general the kinase selectivity profile differed from that of 1 with nearly equipotent inhibition of the wild-type and the Imatinib resistant Abl T(315)I mutant . A potent cellular inhibition of P09619 and cytotoxicity toward EOL-1 cells , a model for idiopathic hypereosinophilic syndrome ( DB09106 ) , are restored or enhanced for selected analogues ( 12b , 14b , and 18b ) . Cytotoxicity was evaluated by using a broad panel of tumor cell lines , with selected analogues displaying mean IC(50) values between 3.6 and 7.1 muM . 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 . In vitro and in vivo interactions between the Q9UBN7 inhibitor ricolinostat ( ACY1215 ) and the irreversible proteasome inhibitor carfilzomib in non-Hodgkin lymphoma cells . Interactions between the Q9UBN7 inhibitor ricolinostat ( ACY1215 ) and the irreversible proteasome inhibitor carfilzomib were examined in non-Hodgkin lymphoma ( Q9NZ71 ) models , including diffuse large B-cell lymphoma ( DLBCL ) , mantle cell lymphoma ( Q8WXI8 ) , and double-hit lymphoma cells . Marked in vitro synergism was observed in multiple cell types associated with activation of cellular stress pathways ( e.g. , P45983 /2 , P27361 /2 , and p38 ) accompanied by increases in DNA damage ( γH2A.X ) , G2-M arrest , and the pronounced induction of mitochondrial injury and apoptosis . Combination treatment with carfilzomib and ricolinostat increased reactive oxygen species ( ROS ) , whereas the antioxidant TBAP attenuated DNA damage , JNK activation , and cell death . Similar interactions occurred in bortezomib-resistant and double-hit DLBCL , Q8WXI8 , and primary DLBCL cells , but not in normal P28906 (+) cells . However , ricolinostat did not potentiate inhibition of chymotryptic activity by carfilzomib . shRNA knockdown of P45983 ( but not Q02750 /2 ) , or pharmacologic inhibition of p38 , significantly reduced carfilzomib-ricolinostat lethality , indicating a functional contribution of these stress pathways to apoptosis . Combined exposure to carfilzomib and ricolinostat also markedly downregulated the cargo-loading protein P54727 . Moreover , P54727 knockdown significantly increased carfilzomib- and ricolinostat-mediated lethality , suggesting a role for this event in cell death . Finally , combined in vivo treatment with carfilzomib and ricolinostat was well tolerated and significantly suppressed tumor growth and increased survival in an Q8WXI8 xenograft model . Collectively , these findings indicate that carfilzomib and ricolinostat interact synergistically in Q9NZ71 cells through multiple stress-related mechanisms , and suggest that this strategy warrants further consideration in Q9NZ71 . Interferon-stimulated transcription and innate antiviral immunity require deacetylase activity and histone deacetylase 1 . The use of histone deacetylase ( HDAC ) inhibitors has revealed an essential role for deacetylation in transcription of IFN-responsive genes . The Q13547 protein associates with both signal transducer and activator of transcription ( P35610 ) 1 and P52630 , and IFN-alpha stimulation induces deacetylation of histone H4 . Inhibition of Q13547 by small interfering RNA ( siRNA ) decreases IFN-alpha responsiveness whereas expression of Q13547 augments the IFN-alpha response , demonstrating that Q13547 modulates IFN-alpha-induced transcription . Importantly , the innate antiviral response is inhibited in the absence of deacetylase activity . The requirement for deacetylase is shared by P01579 transcription response and may represent a general requirement for P35610 -dependent gene expression . 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 . 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 . 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 . Inhibition of cellular growth and migration by suppression of endothelial protein C receptor ( Q9UNN8 ) in lung carcinoma cells . Increasing evidence shows that beyond its role in coagulation , thrombosis interferes with carcinogenesis . Q9UNN8 ( Q9UNN8 ) is a cellular receptor for protein C and activated protein C ( P25054 ) . Such Q9UNN8 -induced signal transduction promotes cancer cell migration , invasion , and angiogenesis and inhibits cancer cell apoptosis . However , its role in lung carcinoma biology is yet to be demonstrated . Here , the recombinant Q9UNN8 siRNA plasmids were constructed to investigate the effects of inhibition of Q9UNN8 on human lung cancer H1299 . Q9UNN8 siRNA led to inhibition of endogenous Q9UNN8 mRNA and protein expression as determined by RT-PCR and Western blotting analysis . Q9UNN8 siRNA significantly inhibited cell growth , blocked entry into the S phase of the cell cycle , and reduced the migration of H1299 cells . Q9UNN8 siRNA also decreases P08253 and cyclin E expression in H1299 cells . In addition , siRNA targeting of Q9UNN8 inhibited the growth of H1299 cells and decreased P53602 in SCID mice tumor models . Taken together , Q9UNN8 was involved in regulating progression of human lung cancer cells . Manipulation of Q9UNN8 expression may be a potential therapeutic strategy for lung cancer . Requirement and epigenetics reprogramming of Nrf2 in suppression of tumor promoter TPA-induced mouse skin cell transformation by sulforaphane . Nrf2 is a transcription factor that plays critical roles in regulating the expression of cellular defensive antioxidants and detoxification enzymes . However , the role of Nrf2 and Nrf2 's epigenetics reprogramming in skin tumor transformation is unknown . In this study , we investigated the inhibitory role and epigenetics of Nrf2 on tumor transformation induced by 12-O-tetradecanoylphorbol-13-acetate ( TPA ) in mouse skin epidermal JB6 ( JB6 P+ ) cells and the anticancer effect of sulforaphane ( SFN ) , an isothiocyanate found in cruciferous vegetables . After five days of treatment , SFN significantly inhibited TPA-induced JB6 cellular transformation and SFN enhanced the nuclear translocation of Nrf2 and increased the mRNA and protein levels of the Nrf2 target genes P09601 , P15559 and P22309 . Knockdown of Nrf2 attenuated the induction of Nrf2 , P09601 and P15559 by SFN , enhanced TPA-induced colony formation and dampened the inhibitory effect of SFN on TPA-induced JB6 transformation . Epigenetics investigation using bisulfite genomic sequencing showed that SFN decreased the methylation ratio of the first 15 CpGs of the Nrf2 gene promoter , which was corroborated by increased Nrf2 mRNA expression . Furthermore , SFN strongly reduced the protein expression of DNA methyltransferases ( P26358 , DNMT3a and DNMT3b ) . SFN also inhibited the total histone deacetylase ( HDAC ) activity and decreased the protein expression of Q13547 , Q92769 , O15379 and P56524 . Collectively , these results suggest that the anti-cancer effect of SFN against TPA-induced neoplastic transformation of mouse skin could involve the epigenetic reprogramming of anti-cancer genes such as Nrf2 , leading to the epigenetic reactivation of Nrf2 and the subsequent induction of downstream target genes involved in cellular protection . [ The inter-individual variability of the response to clopidogrel ] . DB00758 is an anti-platelet aggregation drug with proven efficacy in the prevention of atherothrombotic complications . However , according to studies , between 5 and 20 % of patients have further thrombotic episodes despite treatment with clopidogrel . The principal ex vivo evaluation methods of clopidogrel 's efficacy are platelet aggregometry , and using flow cytometry to measure platelet activation and the quantatitive analysis of P50552 phosphorylation . P50552 analysis is the most selective method for clopidogrel 's effect . These tests show great inter-individual variability in the response to treatment . This variability is such that about a third of treated patients show an " insufficient " response to clopidogrel . The exact causes of this variability have not yet been clearly identified . They could correspond with observer error , inter-individual variability in intestinal absorption or hepatic metabolism , or polymorphism of clopidogrel 's target , the Q9H244 receptor . Accordingly it may be necessary to adapt the dose of clopidogrel or to use an alternative treatment . Failure to respond to clopidogrel would be a risk factor for atherothrombotic complications , but this association has yet to be demonstrated . 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 . Biological and immunological studies of bovine hypothalamic DB05394 . P06850 B ( CRF-B ) is a peptide(s) isolated from bovine hypothalamic extracts by Sephadex G-100 chromatography on the basis of its ability to stimulate secretion of adrenocorticotropin ( DB01285 ) in vitro and in vivo . It is similar in molecular size to the 41-residue ovine CRF ( oCRF ) or rat CRF ( rCRF ) recently elucidated and appears to be their bovine counterpart . Immunoreactivity of CRF-B was examined in homologous radioimmunoassays ( RIAs ) for oCRF or rCRF , using several anti-oCRF and anti-rCRF antibodies . CRF-B cross-reacted well with anti-oCRF antibodies but poorly with anti-rCRF antibodies . Purification of CRF-B with preparative isoelectric focusing yielded four CRF peaks , B-1 ( pH 4.7 ) , B-2 ( pH 5.5 ) , B-3 ( pH 6.3 ) , and B-4 ( pH 7.0 ) , which accounted for 16 , 30 , 46 , and 8 % of the total immunoreactivity , respectively . CRF B-2 , B-3 , and B-4 showed both immunological activity and biological activity in vitro ( cell culture assay ) and in vivo ( Arimura assay ) , whereas CRF B-1 showed only immunoreactivity . Their relative bioactivity/immunoreactivity ratios were 0 ( B-1 ) , 1 ( B-2 ) , 1 ( B-3 ) , and 3 ( B-4 ) . All of these CRF-B subtypes exhibited RIA displacement curves parallel to that for the oCRF standard and coeluted with oCRF on Sephadex G-100 chromatography , which suggests that their molecular modifications are relatively minor . 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 . 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 . 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 . Q9UBN7 -mediated deacetylation of α-tubulin coordinates cytoskeletal and signaling events during platelet activation . The tubulin cytoskeleton plays a key role in maintaining the characteristic quiescent discoid shape of resting platelets . Upon activation , platelets undergo a dramatic change in shape ; however , little is known of how the microtubule system contributes to regulating platelet shape and function . Here we investigated the role of the covalent modification of α-tubulin by acetylation in the regulation of platelet physiology during activation . Superresolution microscopy analysis of the platelet tubulin cytoskeleton showed that the marginal band together with an interconnected web of finer tubulin structures collapsed upon platelet activation with the glycoprotein VI ( Q9HCN6 ) -agonist collagen-related peptide ( CRP ) . Western blot analysis revealed that α-tubulin was acetylated in resting platelets and deacetylated during platelet activation . Tubacin , a specific inhibitor of the tubulin deacetylase Q9UBN7 , prevented tubulin deacetylation upon platelet activation with CRP . Inhibition of Q9UBN7 upregulated tubulin acetylation and disrupted the organization of the platelet microtubule marginal band without significantly affecting platelet volume changes in response to CRP stimulation . Q9UBN7 inhibitors also inhibited platelet aggregation in response to CRP and blocked platelet signaling events upstream of platelet Rho GTPase activation . Together , these findings support a role for acetylation signaling in controlling the resting structure of the platelet tubulin marginal band as well as in the coordination of signaling systems that drive platelet cytoskeletal changes and aggregation . 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 . Urinary pheromones promote P29323 /Akt phosphorylation , regeneration and survival of vomeronasal ( P30518 ) neurons . The G protein-coupled pheromone receptor neurons ( V1R and P30518 ) of the vomeronasal organ ( VNO ) are continually replaced throughout the lifetime of the mouse . Moreover , active signalling of V2Rs via the transient receptor potential 2(TRPC2) channel is necessary for regeneration of receptors , as the TRPC2 null mutant mouse showed a 75 % reduction of V2Rs by the age of two months . Here we describe P30518 mediated signalling in a neuronal line established from vomeronasal stem cells taken from postnatal female mice . Cells were immunoreactive for Galpha(o) and P30518 , whereas V1R and Galpha(i) immunoreactivity could not be detected . Biological ligands ( dilute urine and its protein fractions ) were found to increase proliferation and survival of these neurons . Dilute mouse urine but not artificial urine also induced P29323 , Akt and CREB signalling in a dose dependent way . The volatile fraction of male mouse urine alone was without effect while the fraction containing peptides ( > 5 kDa ) also stimulated P29323 and Akt phosphorylation . The P29323 , Akt and CREB phosphorylation response was sensitive to pertussis toxin , confirming the involvement of P30518 linked Galpha(o) . Dilute mouse urine or its high molecular weight protein fraction increased survival and proliferation of these neurons . Hence , urinary pheromones , which signal important social information via mature neurons , also promote survival and proliferation of their regenerating precursors . These data show that regenerating V2Rs respond to urine and the urinary peptides by activation of the Ras- P29323 and P19957 -Akt pathways , which appear to be important for vomeronasal neural survival and proliferation . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Syntenic conservation between humans and cattle . I . Human chromosome 9 . Bovine X hamster hybrid somatic cells have been used to investigate the syntenic relationship of nine loci in the bovine that have homologous loci on human chromosome 9 . Six loci , P00352 , P05062 , P01031 , P15291 , P06396 , and P02760 , were assigned to the previously identified bovine syntenic group U18 represented by P21399 , whereas the other three loci , P00519 , P00966 , and P11021 , mapped to a new , previously unidentified autosomal syntenic group . Additionally , a secondary locus , P42684 , which cross-hybridized with the P00519 probe , was mapped to bovine syntenic group U1 with the HSA 1 loci P52209 and P06733 . The results predict that P21399 will map proximal to P00352 ; P11021 distal to P02760 and P01031 ; P06396 proximal to P00568 , P00519 , and P00966 on HSA 9 ; P11021 to MMU 2 ; and P02760 and P06396 to MMU 4 . The alkylating carcinogen N-methyl-N'-nitro-N-nitrosoguanidine activates the plasminogen activator inhibitor-1 gene through sequential phosphorylation of p53 by Q13315 and ATR kinases . The alkylating agent MNNG is an environmental carcinogen that causes DNA lesions leading to cell death . We previously demonstrated that MNNG induced the transcriptional activity of the plasminogen activator inhibitor-1 ( P05121 ) gene in a p53-dependent manner . However , the mechanism(s) linking external MNNG stimulation and P05121 gene induction remained to be elucidated . Here , we show that Q13315 and ATR kinases , but not DNA-PK , which participate in DNA damage-activated checkpoints , regulate the phosphorylation of p53 at serine 15 in response to MNNG cell treatment . Using Q13315 -deficient cells , Q13315 was shown to be required for early phosphorylation of serine 15 in response to MNNG , whereas catalytically inactive ATR selectively interfered with late phase serine 15 phosphorylation . In contrast , DNA-PK-deficient cells showed no change in the MNNG-induced serine 15 phosphorylation pattern . In agreement with this , sequential activation of Q13315 and ATR kinases was also required for adequate induction of the endogenous P05121 gene by MNNG . Finally , we showed that cells derived from P05121 -deficient mice were more resistant to MNNG-induced cell death than normal cells , suggesting that p53-dependent P05121 expression partially mediated this effect . Since P05121 is involved in the control of tumor invasiveness , our finding that MNNG induces P05121 gene expression via Q13315 /ATR-mediated phosphorylation of p53 sheds new insight on the role of these DNA damage-induced cell cycle checkpoint kinases . Role of endothelin-1 receptor blockers on hemodynamic parameters and oxidative stress . Endothelin ( ET ) was first isolated and described by Yanagisawa et al. and has since been described as one of the most potent known vasoconstrictor compounds . ET-1 mediates its effects via two types of receptors , P25101 and ETB , which are expressed in the vascular smooth muscle cells , endothelial cells , intestines and brain . Secretion of ET-1 results in long-lasting vasoconstriction , increased blood pressure and , in turn , overproduction of free radicals . As dysregulation of the endothelin system is an important factor in the pathogenesis of several diseases including atherosclerosis , hypertension and endotoxic shock , the P25101 and ETB receptors are attractive therapeutic targets for treatment of these disorders . The biosynthesis and release of ET-1 are regulated at the transcriptional level . Studies have shown that p38MAP kinase , nuclear factor kappaB ( NF-kappaB ) , PKC/ P29323 and JNK/c-Jun all take part in the ROS-activated production of ET-1 . Furthermore , administration of ET(A) significantly reduces the generation of free radicals . However , treatment with ETB receptor blockers does not elicit the same effect . Therefore , the effects of endothelin receptor blockers on blood pressure and the generation of free radicals remain debatable . This review summarizes recent investigations into the role of endothelin receptor blockers with respect to the modulation of hemodynamic parameters and the generation of free radicals . Identifying new targets in leukemogenesis using computational approaches . There is a need to identify novel targets in Acute Lymphoblastic Leukemia ( ALL ) , a hematopoietic cancer affecting children , to improve our understanding of disease biology and that can be used for developing new therapeutics . Hence , the aim of our study was to find new genes as targets using in silico studies ; for this we retrieved the top 10 % overexpressed genes from Oncomine public domain microarray expression database ; 530 overexpressed genes were short-listed from Oncomine database . Then , using prioritization tools such as ENDEAVOUR , P30518 and TOPPGene online tools , we found fifty-four genes common to the three prioritization tools which formed our candidate leukemogenic genes for this study . As per the protocol we selected thirty training genes from PubMed . The prioritized and training genes were then used to construct STRING functional association network , which was further analyzed using cytoHubba hub analysis tool to investigate new genes which could form drug targets in leukemia . Analysis of the STRING protein network built from these prioritized and training genes led to identification of two hub genes , Q15796 and P50750 , which were not implicated in leukemogenesis earlier . Filtering out from several hundred genes in the network we also found O00255 , Q13547 and P06239 genes , which re-emphasized the important role of these genes in leukemogenesis . This is the first report on these five additional signature genes in leukemogenesis . We propose these as new targets for developing novel therapeutics and also as biomarkers in leukemogenesis , which could be important for prognosis and diagnosis . 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 . 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 . Thrombin and activated protein C inhibit the expression of secretory group IIA phospholipase A(2) in the P01375 -activated endothelial cells by Q9UNN8 and P25116 dependent mechanisms . INTRODUCTION : Thrombin and tumor necrosis factor ( P01375 ) -alpha up-regulate the expression of proinflammatory molecules in human umbilical vein endothelial cells ( HUVECs ) . However , activated protein C ( P25054 ) down-regulates the expression of the same molecules . The expression level of secretory group IIA phospholipase A(2) ( sPLA(2)-IIA ) is known to be elevated in inflammatory disorders including in sepsis . Here , we investigated the effects of P25054 and thrombin on the expression of sPLA(2)-IIA and extracellular signal-regulated kinase ( P29323 ) in HUVECs . MATERIALS AND METHODS : The expression level of sPLA(2)-IIA was quantitatively measured by an enzyme-linked-immunosorbent-assay following stimulation of HUVECs with either thrombin or P01375 in the absence and presence of the phosphatidylinositol 3-kinase ( P19957 -kinase ) inhibitor LY294002 and the cholesterol-depleting drug methyl-beta-cyclodextrin ( MbetaCD ) . RESULTS AND CONCLUSIONS : Thrombin had no effect on the expression of sPLA(2)-IIA in HUVECs , however , P01375 potently induced its expression . The prior treatment of cells with P25054 inhibited expression of sPLA(2)-IIA through the Q9UNN8 -dependent cleavage of P25116 . Further studies revealed that if HUVECs were pretreated with the zymogen protein C to occupy Q9UNN8 , thrombin also inhibited the P01375 -mediated expression of sPLA(2)-IIA through the cleavage of P25116 . The Q9UNN8 -dependent cleavage of P25116 by both P25054 and thrombin increased the phosphorylation of P29323 1/2 . Pretreatment of cells with either LY294002 or MbetaCD abolished the inhibitory activity of both P25054 and thrombin against sPLA(2)-IIA expression , suggesting that the protein C occupancy of Q9UNN8 confers a P19957 -kinase dependent protective activity for thrombin such that its cleavage of the lipid-raft localized P25116 inhibits the P01375 -mediated expression of sPLA(2)-IIA in HUVECs .

MH_train_1113

MH_train_1113

MH_train_1113

interacts_with DB00243?

multiple_choice

Identification of a histamine H4 receptor on human eosinophils -- role in eosinophil chemotaxis . Eosinophils are recruited to sites of inflammation via the action of a number of chemical mediators , including Q15004 , leukotrienes , eotaxins , ECF-A and histamine . Although many of the cell-surface receptors for these mediators have been identified , histamine-driven chemotaxis has not been conclusively attributed to any of the three known histamine receptor subtypes , suggesting the possibility of a 4th histamine-responsive receptor on eosinophils . We have identified and cloned a novel G protein-coupled receptor ( GPCR ) , termed Q9H3N8 , from an P05113 stimulated eosinophil cDNA library which is homologous to the human histamine H3 receptor , both at the sequence and gene structure level . Expression data indicates that Q9H3N8 is predominantly expressed in peripheral blood leukocytes , with lower expression levels in spleen , testis and colon . Ligand-binding studies using Q9H3N8 expressed in P29320 -293Galpha15 cells , demonstrates specific binding to histamine with a Kd of 3.28 +/- 0.76 nM and possesses a unique rank order of potency against known histaminergic compounds in a competitive ligand-binding assay ( histamine > clobenpropit > iodophenpropit > thioperamide > R-alpha-methylhistamine > cimetidine > DB06691 ) . We have therefore termed this receptor human histamine H4 . Chemotaxis studies on isolated human eosinophils have confirmed that histamine is chemotactic and that agonists of the known histamine receptors ( H1 , H2 , and H3 ) do not induce such a response . Furthermore , studies employing histamine-receptor antagonists have shown an inhibition of chemotaxis only by the H3 antagonists clobenpropit and thioperamide . Since these compounds are also antagonists of hH4 we postulate that the receptor mediating histaminergic chemotaxis is this novel histamine H4 receptor . Self-assembly of P29320 cell-secreted ApoE particles resembles ApoE enrichment of lipoproteins as a ligand for the P01130 -related protein . Recent studies have shown that the lipidation and assembly state of apolipoprotein E ( apoE ) determine receptor recognition and amyloid-beta peptide ( Abeta ) binding . We previously demonstrated that apoE secreted by P29320 cells stably expressing apoE3 or apoE4 ( P29320 -apoE ) binds Abeta and inhibits Abeta-induced neurotoxicity by an isoform-specific process that requires apoE receptors . Here we characterized the structure of P29320 -apoE assemblies and determined their receptor binding specificity . By chromatography , P29320 -apoE elutes in high molecular mass fractions and is the size of plasma HDL , consistent with a multiprotein assembly . No lipid was associated with these apoE assemblies . Several methods for analyzing receptor binding indicate that P29320 -apoE is a ligand for low-density lipoprotein ( LDL ) receptor-related protein ( Q14764 ) but not the P01130 . This suggests that self-assembly of apoE may induce a functional conformation necessary for binding to Q14764 . Our results indicate that , in addition to lipid content , the assembly state of apoE influences Abeta binding and receptor recognition . A novel strategy for the generation of angiostatic kringle regions from a precursor derived from plasminogen . In this study we have explored the feasibility of generating angiostatin by incorporating an endoproteolytic furin cleavage site into plasminogen to allow conversion of the precursor molecule into an angiostatic active P04264 -3 fragment . We show that secretable angiostatin can be successfully generated from cells infected with adenovirus carrying the furin-mutated plasminogen ( AdmuthPlgK3 ) . Supernatant from cells transduced with AdmuthPlagK3 inhibits tube formation and proliferation and migration of human umbilical vein endothelial cells with an efficiency similar to that of supernatant from cells infected with adenovirus expressing kringle 1-3 of plasminogen ( AdK1-3 ) . Administration of AdmuthPlgK3 and AdK1-3 in mice results in significantly decreased endothelial cell infiltration in P15692 -embedded Matrigel plugs . Treatment with AdmuthPlgK3 and AdK1-3 exerts strong antitumoral effect in models of hepatocellular carcinoma and Lewis lung cancer . This antitumor effect was associated with decreased microvessel density in the tumors . Taken together , our data demonstrate that angiostatin endowed with strong antiangiogenic and antitumor effects can be released from a furin-mutated plasminogen acting as a precursor . This strategy may have potential to develop angiostatic anti-cancer therapies . 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 . Molecular and genotypic characterization of human thyroid follicular cell carcinoma-derived cell lines . OBJECTIVE : Our aim was to characterize the molecular and genotypic profile of eight thyroid carcinoma-derived cell lines- Q9ULQ1 , FB2 , B- Q9HC77 , P04264 , XTC-1 , C643 , 8505C , and Hth74-in order to use them as in vitro models of thyroid carcinogenesis . DESIGN : We evaluated the expression of five thyroid-specific genes ( Tg , TSHr , P07202 , Q06710 , and Q15669 -1 ) to establish the cell lineage and to assess the differentiation status of each of the cell lines . We screened for mutations in the most relevant oncogenes/tumor suppressor genes affected in thyroid carcinogenesis : DB01367 , P15056 , P35222 , and P04637 along with P07949 /PTC rearrangements . Considering the putative relevance in general carcinogenesis , we have also studied other molecules such as P00533 , PI3K , RAF-1 , and P10828 . To determine the genetic identity of the cell lines , we performed genotypic analysis . MAIN OUTCOME : The panel of cell lines we have studied displayed activation of several oncogenes ( P15056 , DB01367 , P07949 /PTC ) and inactivation of tumor suppressor genes ( P04637 ) known to be important for thyroid carcinogenesis . Two of the cell lines- Q9ULQ1 and FB2-shared the same genotypic profile , probably representing clones of an ancestor cell line ( Q9ULQ1 ) . CONCLUSION : Due to their different molecular alterations , these cell lines represent a valuable tool to study the molecular mechanisms underlying thyroid carcinogenesis . We suggest that genotypic analyses should be included as a routine procedure to guarantee the uniqueness of each cell line used in research . Modulation of expression in BEAS-2B airway epithelial cells of α-L-fucosidase A1 and A2 by Th1 and Th2 cytokines , and overexpression of α-L-fucosidase 2 . Chronic Th2-driven airway inflammation with excessive mucus production occurs in asthma . The regulation of P04066 and Q9BTY2 gene expression and enzyme activity in response to asthma-associated Th2 cytokines and , for contrast , Th1 cytokine IFN-γ , were investigated in a human airway cell line . BEAS-2B cells were supplemented with Th2-derived cytokines ( P35225 , P05112 , P05113 ) or/and IFN-γ . RNA and cell supernatants from stimulated and unstimulated cells were collected over a period of 3 h . Alpha-L-fucosidase A1 and A2 gene expression were assessed using real time RT-PCR , while enzymatic activities were measured using a fluorescent assay . To characterise α-L-fucosidase A2 , CHO- P04264 and BEAS-2B cell lines were transiently transfected , the Q9BTY2 gene was overexpressed , and the protein was immunoprecipitated . The transcription of P04066 was upregulated ( p < 0.01 ) in response to IFN-γ , suggesting that P04066 transcription and fucosidase activity are regulated in a Th1-dependent manner . The gene expression was the highest for 30 min after IFN-γ stimulation ( > twofold induction ) , whereas secreted enzyme activity in BEAS-2B cells was significantly increased 1 h after IFN-γ addition . P05112 , P05113 and P35225 had no effect on P04066 and Q9BTY2 expression and activity . The IFN-γ-induced increase in expression and activity was repressed by the presence of the Th2 cytokine P05113 . Enzymatically active α-L-fucosidase 2 was immunoprecipitated from BEAS-2B cells , with highest activity at pH 4.9 . P35225 , P05112 and P05113 have no effect on the expression of P04066 and Q9BTY2 , but its expression is upregulated by IFN-γ , a Th1 cytokine . Active α-L-fucosidase 2 was overexpressed in BEAS-2B cells . 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 . Inward rectifier potassium currents as a target for atrial fibrillation therapy . Subunits of inwardly rectifying potassium channels ( Kir ) are expressed in many different tissues of the human body . Inward rectifier currents expressed in the heart are constituted by pore-forming alpha-subunits of Kir2 , Kir3 , and Kir6 subfamilies . Characteristic properties of inward rectifiers comprise small outward conductances that nevertheless are important to terminal repolarization of cardiac action potentials . There is considerable difference in the regional expression of cardiac Kir channels , and subunits are additionally regulated by specific disease conditions . Resulting changes facilitate occurrence and persistence of atrial fibrillation ( AF ) . For instance , upregulation of Kir2.1 protein and resultant current I P04264 is a hallmark of AF-related ionic remodeling . Increased I P04264 helps to stabilize atrial rotors , and current inhibition has accordingly been suggested as an antiarrhythmic approach for AF therapy . But there are caveats to I P04264 inhibition per se , and there is no specific inhibitor of Kir2 channels . Modulation of I P04264 rectification properties seems theoretically interesting for manipulation of Kir2 currents as an antiarrhythmic approach . Kir3-based muscarinic currents ( I KACh ) are functionally upregulated during AF through increased constitutive activity ( passing current in the absence of an agonist ) . Upregulated I KACh supports sustenance of the arrhythmia . There is considerable intraatrial diversity in the expression of underlying Kir3.1/Kir3.4 subunits , but atrial-specific localization makes inhibition of this current a potentially interesting antiarrhythmic target devoid of ventricular side effects . Experimental studies of specific inhibitors indicate efficacy in various disease models . The role of I KATP remodeling under AF conditions has not been extensively studied , but present evidence indicates current downregulation and modulation of Q14654 seems less promising than that of other inward rectifiers . Lectin-like oxidized P01130 -1 ( P78380 ) supports adhesion of mononuclear leukocytes and a monocyte-like cell line THP-1 cells under static and flow conditions . Adhesion of mononuclear leukocytes to vascular endothelial cells appears one of the initial steps in the process of atherogenesis and inflammation . We examined if P78380 , an endothelial scavenger receptor with C-type lectin-like structure , can support adhesion of mononuclear leukocytes . Under a static condition , CHO- P04264 cells stably expressing P78380 showed more prominent adhesion of human peripheral blood mononuclear leukocytes and THP-1 cells than untransfected CHO- P04264 cells , in a temperature-independent fashion . Mononuclear leukocytes also adhered to plastic plates precoated with recombinant soluble P78380 extracellular domain . A neutralizing anti- P78380 monoclonal antibody , as well as oxidized low-density lipoprotein , significantly blocked adhesion of THP-1 cells to CHO- P04264 cells overexpressing P78380 and bovine aortic endothelial cells . Under a flow condition , increased numbers of THP-1 cells showed rolling with reduced velocities on P78380 -expressing CHO- P04264 cells , compared with those on untransfected CHO- P04264 cells . Taken together , P78380 can work as a cell surface receptor for mononuclear leukocytes under both static and flow conditions . 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 . 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 . 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 . 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 . 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 . 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 . Dynamic assembly of the urokinase-type plasminogen activator signaling receptor complex determines the mitogenic activity of urokinase-type plasminogen activator . The urokinase-type plasminogen activator ( uPA ) receptor ( Q03405 ) functions in concert with co-receptors , including integrins , P21462 -like receptor-1/lipoxin A4 receptor , and the epidermal growth factor receptor ( P00533 ) , to initiate cell signaling . Q03405 co-receptors may be dynamically organized into a multiprotein signaling receptor complex . In Chinese hamster ovary- P04264 ( CHO- P04264 ) cells , uPA-binding to Q03405 activates P29323 / Q96HU1 kinase , even though these cells do not express the P00533 ; however , when CHO- P04264 cells are transfected to express the P00533 , P29323 activation becomes P00533 -dependent . In this study , we demonstrate that P29323 activation in response to uPA follows equivalent biphasic kinetics in P00533 -expressing and -deficient CHO- P04264 cells . In both cell types , the response is pertussis toxin-sensitive ; however , uPA promotes cell proliferation exclusively in the P00533 -expressing cells . uPA-induced mitogenic activity requires activation of both STAT5b and P29323 . STAT5b was tyrosine-phosphorylated , in response to uPA , only in P00533 -expressing cells . uPA-induced cell proliferation was blocked by dominant-negative Q02750 , dominant-negative STAT5b , and by expression of an P00533 that is mutated at DB00135 -845 , which is essential for STAT5b activation . In two cell culture models of uPA-stimulated breast cancer growth , MDA-MB 468 cells treated with uPA and MCF-7 cells treated with uPA-plasminogen activator inhibitor-1 complex , proliferation was completely inhibited when P00533 expression or activity was blocked . We conclude that expression and assembly of Q03405 co-receptors in a specific cell type determines the response to uPA . The P00533 selectively cooperates with Q03405 to mediate mitogenesis . 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 . 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 . Characterization of a thyroiditis-inducing thyroglobulin-specific T-cell clone restricted by the H-2 molecule of a low responder mouse strain . We established a thyroglobulin ( Tg ) -specific , thyroiditis-inducing T-cell clone , B12G , from B6C3F1 mice by the immunization of mouse Tg with lipopolysaccharide ( LPS ) from Klebsiella strain LEN ( O3: P04264 ) . B12G was Thy-1.2+ , CD3+ , P01730 + , P05107 + , and CD8- , and could transfer thyroiditis to recipient mice after in vitro stimulation with mouse or bovine Tg . Histological examination showed severe thyroiditis with predominant infiltrations of polymorphonuclear cells ; few mononuclear cells were observed . B12G proliferated in response to bovine , mouse , porcine , and rat Tg in the presence of irradiated spleen cells , but did not respond to chicken or human Tg . H-2b , a low-responder haplotype of experimental autoimmune thyroiditis , governed the response of the clone to Tg . B12G produced interleukin-4 ( P05112 ) and P05231 , but not P60568 or interferon-gamma ( P01579 ) , on stimulation with mouse Tg . These findings were different from characteristics of previously reported Tg-specific T-cell clones from high-responder mice in terms of epitope specificity and cytokine production pattern , raising the possibility that the specificities and functions of T cells involved in the development of autoimmune thyroiditis in low-responder mice differ from those in high responders . 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 . 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 . A functional variant of the dopamine D3 receptor is associated with risk and age-at-onset of essential tremor . Familial essential tremor ( ET ) , the most common inherited movement disorder , is generally transmitted as an autosomal dominant trait . A genome-wide scan for ET revealed one major locus on chromosome 3q13 . Here , we report that the Ser9Gly variant in the dopamine D(3) receptor gene ( P35462 ) , localized on 3q13.3 , is associated and cosegregates with familial ET in 23 out of 30 French families . Sequencing revealed no other nonsynonymous variants in the P35462 -coding sequence and in the first 871 bp of the 5' flanking region . Moreover , DB00145 -9 homozygous patients presented with more severe and/or earlier onset forms of the disease than heterozygotes . A replication study comparing 276 patients with ET and 184 normal controls confirmed the association of the DB00145 -9 variant with risk and age-at-onset of ET . In human embryonic kidney ( P29320 ) 293-transfected cells , the DB00145 -9 variant did not differ from the DB00133 -9 variant with respect to glycosylation and to anterograde and retrograde trafficking , but dopamine had an affinity that was four to five times higher . With the DB00145 -9 variant , the dopamine-mediated DB02527 response was increased , and the mitogen-associated protein kinase ( MAPK ) signal was prolonged , as compared with the DB00133 -9 variant . The gain-of-function produced by the DB00145 -9 variant may explain why drugs active against tremor in Parkinson 's disease ( PD ) are usually not effective in the treatment of ET and suggests that P35462 partial agonists or antagonists should be considered as novel therapeutic options for patients with ET . Targeting of plasmid DNA-lipoplexes to cells with molecules anchored via a metal chelator lipid . BACKGROUND : The ability to deliver plasmid DNA ( pDNA ) to specific cells in vivo is crucial for achieving efficient targeted transfection with nonviral vectors . We previously used stealth liposomes containing the chelator lipid 3 ( nitrilotriacetic acid ) -ditetradecylamine ( NTA(3)-DTDA ) to target delivery of antigen and cytokines to immune cells in vivo . In the present study , we utilized liposomes containing NTA(3)-DTDA and the ionizable aminolipid 1,2-dioleoyl-3-dimethyl-ammonium-propane ( DODAP ) to incorporate pDNA into complexes for targeting to cells . METHODS : Liposomes containing DODAP , NTA(3)-DTDA and helper lipids were acidified ( pH 5.5 ) and mixed with pDNA to form complexes . These lipoplexes were neutralized and engrafted with DB00117 -tagged molecules for targeting to extracellular receptors . Targeted transfection efficiency was assessed using the enhanced green fluorescent protein reporter gene . RESULTS : Initial transfections of P29320 -293 cells using a DB00117 -tagged peptide ( P24752 ) related to the DB00125 -rich motif of HIV-1 TAT protein resulted in a low transfection efficiency ( < 2.5 % ) . Optimization of the lipid formulation and use of an endosome-destabilizing peptide and inhibitor of DNase II , increased transfection approximately 20-fold . These lipoplexes are approximately 250 nm in diameter , and transfection efficiencies were : approximately 50 % for P29320 -293 cells targeted with lipoplexes containing pEGFP-N1 and engrafted with P24752 , and 30-40 % for HepG2 cells targeted with lipoplexes engrafted with a peptide specific for the P15692 receptor Flt-1 . CONCLUSIONS : The results show that DODAP-containing lipoplexes incorporating NTA(3)-DTDA enable the engraftment of targeting molecules and the effective targeting of pDNA to cells in serum-containing media , resulting in efficient transgene expression . The strategy may provide a convenient approach for targeting pDNA to cells in vivo in therapeutic applications . 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 . 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 . 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 . Functional analysis of the human D2 dopamine receptor missense variants . The human dopamine D2 receptor gene ( P14416 ) has three polymorphic variants that predict the amino acid substitutions Val96 --> Ala , Pro310 --> DB00133 , and Ser311 --> DB00151 in the receptor protein . We have investigated the ligand binding and signal transduction properties of these human D2 receptor variants by stably expressing them in cultured mammalian cells . The Cys311 and Ser310 variants of the human D2 receptor , which involve substitutions located in the third cytoplasmic loop , were markedly less effective in inhibiting DB02527 synthesis than the most prevalent form ( Pro310 , Ser311 ) . Despite this difference , the Cys311 and Ser310 variants couple to G proteins in CHO- P04264 ( Chinese hamster ovary ) cells . The impairment of the Cys311 and Ser310 variants to inhibit DB02527 levels thus appears to result from a reduced ability of those variant receptors to activate the appropriate Gi-like protein . The demonstration of substantial functional differences between P14416 gene variants found in the human population might have important pharmacological implications given the widespread use of D2 receptor blocking drugs in the treatment of schizophrenia and other psychiatric disorders . 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 . The antimalarial drug mefloquine inhibits cardiac inward rectifier K+ channels : evidence for interference in PIP2-channel interaction . The antimalarial drug mefloquine was found to inhibit the KATP channel by an unknown mechanism . Because mefloquine is a Cationic amphiphilic drug and is known to insert into lipid bilayers , we postulate that mefloquine interferes with the interaction between PIP2 and Kir channels resulting in channel inhibition . We studied the inhibitory effects of mefloquine on Kir2.1 , Kir2.3 , Kir2.3(I213L) , and Kir6.2/SUR2A channels expressed in P29320 -293 cells , and on IK1 and Q14654 from feline cardiac myocytes . The order of mefloquine inhibition was Kir6.2/SUR2A ≈ Kir2.3 ( IC50 ≈ 2 μM ) > Kir2.1 ( IC50 > 30 μM ) . Similar results were obtained in cardiac myocytes . The Kir2.3(I213L) mutant , which enhances the strength of interaction with PIP2 ( compared to WT ) , was significantly less sensitive ( IC50 = 9 μM ) . In inside-out patches , continuous application of PIP2 strikingly prevented the mefloquine inhibition . Our results support the idea that mefloquine interferes with PIP2-Kir channels interactions . 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 .

MH_train_1114

MH_train_1114

MH_train_1114

interacts_with DB00054?

multiple_choice

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 . Enhanced P11274 - P00519 kinase inhibition does not result in increased inhibition of downstream signaling pathways or increased growth suppression in CML progenitors . The therapeutic success of imatinib in chronic myeloid leukemia ( CML ) is hampered by persistence of malignant stem cells . We investigated whether nilotinib , a more potent P11274 - P00519 kinase inhibitor could target CML primitive progenitors more effectively than imatinib . CML and normal progenitor cells were cultured with nilotinib or imatinib in growth factor supplemented medium . DB04868 inhibited P11274 - P00519 kinase activity at lower concentrations than imatinib . DB04868 inhibited mitogen-activated protein kinase ( MAPK ) , AKT and P42229 phosphorylation in CML P28906 (+) cells in the absence of growth factors ( GFs ) , but did not suppress AKT and P42229 activity , and resulted in increased MAPK activity , in the presence of GFs . DB04868 and imatinib resulted in similar suppression of CML primitive and committed progenitors in long-term culture-initiating cell and colony-forming cell assays . Inhibition of progenitor growth was related to marked reduction in proliferation , but only a modest increase in apoptosis . DB04868 did not show increased efficacy in reducing nondividing CML progenitors compared with imatinib . These results indicate that more potent tyrosine kinase inhibitors by themselves will not be more effective in eliminating CML progenitors than imatinib and that additional mechanism required for maintenance of malignant stem cells need to be identified to improve targeting of leukemia stem cells . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . A radiation hybrid map of the P38398 region of chromosome 17q12-q21 . The chromosomal region 17q12-q21 contains a gene ( P38398 ) conferring susceptibility to early-onset familial breast and ovarian cancer . An 8000-rad radiation-reduced hybrid ( RH ) panel was constructed to provide a resource for long-range mapping of this region . A large fraction of the hybrids ( approximately 90 % ) retained detectable human chromosome 17 sequences . The complete panel of 76 hybrids was scored for the presence or absence of 22 markers from this chromosomal region , including 14 cloned genes , seven microsatellite repeats , and one anonymous DNA segment . Statistical analysis of the marker retention data employing multipoint methods provided both comprehensive and framework maps of this chromosomal region , including distance estimates between adjacent markers . The comprehensive RH map includes 17 loci and spans 179 cRays(8000) . Likelihood ratios of at least 1000:1 support the 10-locus framework order : cen-D17S250- P04626 - ( P10827 , P11388 ) -D17S855- P01298 -D17S190- P10636 - P05106 ++ +- Q8TDV2 -D17S588-tel . The order obtained from RH mapping , when used in conjunction with other methods , will be useful in linkage analysis of breast cancer families and will facilitate the development of a physical map of this region . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Extramedullary tumor as presentation of leukemia : establishment of a new human P08514 - and P05106 -positive leukemia cell line . A 25-year-old man noted swelling of the right cervical lymph nodes in October 1983 . Diagnosis of malignant lymphoma was made on the basis of pathological examination of biopsies . Despite both chemotherapy and irradiation treatment , blast cells appeared in the peripheral blood and bone marrow in April 1984 . Immunophenotypic analysis demonstrated that the blasts in the patient 's peripheral blood expressed P15144 , P20138 , CD41a , and no markers for T or B lymphocytes , suggesting that he had been suffering from megakaryocytic sarcoma . We established a new cell line derived from the blasts in the peripheral blood , designated KH184 . KH184 cells expressed glycoprotein ( GP ) Ib ( CD42b ) and P08514 /IIIa ( CD41a ) , while platelet peroxidase ( P50336 ) activity was negative in an ultrastructural study . Both Northern blot and flow cytometric analysis of surface antigens and DNA content revealed that treatment with 12-O-tetradecanoylphorbol 13-acetate ( TPA ) did not induce the maturation of these cells . Various cytokines such as interleukin 3 ( P08700 ) , interleukin 6 ( P05231 ) , and leukemia inhibitory factor ( P15018 ) had no effect in promoting the growth of KH184 cells . KH184 cells expressing CD41a seem to possess unusual characteristics . KH184 cells , human P08514 - and P05106 -positive leukemia cells , which lack response to TPA-induced differentiation , provide a new and unique model for the characterization of factors that are implicated in the terminal differentiation of megakaryocytes , and should aid in studies of the mechanism underlying the occurrence of megakaryocytic sarcoma . [ 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 . Contribution of platelets and the vessel wall to the antithrombotic effects of a single bolus injection of Fab fragments of the antiplatelet P08514 /IIIa antibody 7E3 in a canine arterial eversion graft preparation . The contribution of platelets and the vessel wall to the antithrombotic effects of a single intravenous bolus injection of 0.8 mg/kg Fab fragments of the monoclonal antiplatelet glycoprotein IIb/IIIa receptor antibody DB00054 ( 7E3-Fab ) , combined with continuous heparin anticoagulation ( 100 U/kg bolus and 50 U/kg per hour ) , was studied in a canine preparation consisting of an everted ( inside out ) carotid arterial segment that had been inserted into a transected femoral artery . In all 6 control dogs without antibody , persistent or transient eversion graft occlusion occurred during an initial 2-hour observation period , and 5 of the 6 grafts were occluded at 24 hours . In 6 dogs given 7E3-Fab 24 hours before receiving an everted carotid artery segment from a donor dog , cyclic occlusion and reflow occurred in all dogs , whereas the grafts were patent at the end of a 2-hour observation period in 5 of the 6 dogs ( P = .056 versus control ) . When transferred back to the donor dogs , the patient eversion segments showed brief periods of cyclic occlusion and reflow within 2 hours in 3 of 5 dogs ( P = .034 versus control ) , whereas all of the 5 eversion segments were patent at 24 hours ( P < .005 versus control ) . ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 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 . 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 ) 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 . 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 . [ New immunological weapons for medicine in the 21st Century : biological therapy based on the use of the latest generation monoclonal antibodies ] . The fusion of a murine B cell and a myeloma cell generates a hybridoma that produces monoclonal antibody ( mAb ) . These murine mAb induce the HAMA ( human anti-mouse antibodies ) response . Murine mAb have been modified by genetic engineering , producing molecules with a higher proportion of human protein . At present , chimeric , humanized and fully human mAb are available . mAb block interactions between target molecules and their ligands or trigger the lyses of mAb-coated tumor cells . Numerous mAb have been developed using the recombinant DNA technology and several are available in the market . DB00072 , against P04626 /neu , is useful in breast cancer ; rituximab , against P11836 in B lymphocytes is useful in lymphoma ; alemtuzumah , against P31358 is used in lymphoma and leukemia ; daclizumab and basiliximab block the P60568 receptor interaction and reduce acute rejection in kidney transplantation ; abciximab , an antagonist of P08514 /IIIa platelet receptor , is used in patients undergoing acute coronary syndromes . In autoimmunity diseases , blocking tumor necrosis factor by infliximab and adalimumab has demonstrated excellent results . Thus , infliximab is useful in the treatment of rheumatoid arthritis ( RA ) , Crohn 's disease and ulcerative colitis while adalimumab is the first fully human mAb available for RA . DB00065 and adalimumab reduce signs and symptoms in RA and they also interfere with progression of joint damage . Finally , the direct benefits of antagonist treatment can occur at the expense of a major adverse effect in some other biological function . 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 . 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 . 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 . 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 . 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 . 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 . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . 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 . 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 . [ Adhesion molecules in urologic tumors ] . Adhesion molecules play an important role in organogenesis , would healing , inflammation , and progression of malignant tumors . Three major classes of adhesion molecules may be discriminated by function : ( a ) calcium-dependent homotypic adhesion molecules ( e.g. cadherins ) , ( b ) substrate adhesion molecules ( e.g. integrins ) and ( c ) heterotypic adhesion molecules ( e.g. P05362 ) . Molecules of each of the three classes have been identified in urologic tumors . Results of research on substrate adhesion molecules and heterotypic adhesion molecules have not yet led to new clinical concepts . In contrast , loss of P12830 in tumors of the bladder and prostate has been clearly associated with de-differentiation of tumors and diminished survival of patients . Loss of another adhesion molecule , C- P62158 , has been observed in prostate cancer . This has led to new therapeutic approaches , which are in an experimental stage at present . It may be expected that , in the future , new therapeutic concepts will be based on research on adhesion molecules in urologic tumors . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Purification and characterization of platelet aggregation inhibitors from snake venoms . Proteins that inhibit glycoprotein ( GP ) IIb/IIIa mediated platelet aggregation have been purified from the venom of two snake species . A small platelet aggregation inhibitor ( p1.AI ) , multisquamatin ( Mr = 5,700 ) , was purified from Echis multisquamatus venom by hydrophobic interaction HPLC and two steps on C18 reverse phase HPLC . A larger p1.AI , contortrostatin ( Mr = 15,000 ) , was purified by a similar HPLC procedure from the venom of Agkistrodon contortrix contortrix . Both p1.AIs inhibit ADP-induced human , canine and rabbit platelet aggregation using platelet rich plasma ( PRP ) . Multisquamatin has an IC50 of 97 nM , 281 nM and 333 nM for human , canine and rabbit PRP , respectively . Contortrostatin has an IC50 of 49 nM , 120 nM and 1,150 nM for human , canine and rabbit PRP , respectively . In a competitive binding assay using 125I- DB00054 ( a monoclonal antibody to P08514 /IIIa that inhibits platelet aggregation ) both contortrostatin and multisquamatin demonstrated P08514 /IIIa specific binding to human and canine platelets . The IC50 for contortrostatin displacement of 7E3 binding to human and canine P08514 -/IIIa is 27 nM and 16 nM , respectively and for multisquamatin it is 3 nM and 63 nM , respectively . Our results indicate that both p1.AIs inhibit platelet aggregation by binding with high affinity to P08514 /IIIa . Using ImageJ for the quantitative analysis of flow-based adhesion assays in real-time under physiologic flow conditions . This article intends to close the gap between the abundance of regular articles focusing on adhesive mechanisms of cells in a flow field and purely technical reports confined to the description of newly developed algorithms , not yet ready to be used by users without programming skills . A simple and robust method is presented for analysing raw videomicroscopic data of flow-based adhesion assays using the freely available public domain software ImageJ . We describe in detail the image processing routines used to rapidly and reliably evaluate the number of adherent and translocating platelets in videomicroscopic recordings . The depicted procedures were exemplified by analysing platelet interaction with immobilized P04275 and fibrinogen in flowing blood under physiological wall shear rates . Neutralizing GPIbalpha function reduced shear-dependent platelet translocation on P04275 and abolished firm platelet adhesion . DB00054 , Tirofiban and DB00063 completely inhibited P08514 /IIIa-dependent stable platelet deposition on fibrinogen . The presented method to analyse videomicroscopic recordings from flow-based adhesion assays offers the advantage of providing a simple and reliable way to quantify flow-based adhesion assays , which is completely based on ImageJ and can easily be applied to study adhesion mechanisms of cells in non-fluorescent modes without the need to deviate from the presented protocol . 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 . 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 . Complementary roles for receptor clustering and conformational change in the adhesive and signaling functions of integrin alphaIIb beta3 . Integrin alphaIIb beta3 mediates platelet aggregation and " outside-in " signaling . It is regulated by changes in receptor conformation and affinity and/or by lateral diffusion and receptor clustering . To document the relative contributions of conformation and clustering to alphaIIb beta3 function , alphaIIb was fused at its cytoplasmic tail to one or two P62942 repeats ( FKBP ) . These modified alphaIIb subunits were expressed with beta3 in CHO cells , and the heterodimers could be clustered into morphologically detectable oligomers upon addition of AP1510 , a membrane-permeable , bivalent FKBP ligand . Integrin clustering by AP1510 caused binding of fibrinogen and a multivalent ( but not monovalent ) fibrinogen-mimetic antibody . However , ligand binding due to clustering was only 25-50 % of that observed when alphaIIb beta3 affinity was increased by an activating antibody or an activating mutation . The effects of integrin clustering and affinity modulation were additive , and clustering promoted irreversible ligand binding . Clustering of alphaIIb beta3 also promoted cell adhesion to fibrinogen or P04275 , but not as effectively as affinity modulation . However , clustering was sufficient to trigger fibrinogen-independent tyrosine phosphorylation of pp72(Syk) and fibrinogen-dependent phosphorylation of pp125( Q05397 ) , even in non-adherent cells . Thus , receptor clustering and affinity modulation play complementary roles in alphaIIb beta3 function . Affinity modulation is the predominant regulator of ligand binding and cell adhesion , but clustering increases these responses further and triggers protein tyrosine phosphorylation , even in the absence of affinity modulation . Both affinity modulation and clustering may be needed for optimal function of alphaIIb beta3 in platelets . 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 . 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 . Survey of early disapearance of P11274 / P00519 fusion transcript after allogeneic or autologous stem cell transplantation for chronic myelogenous leukemia . The detection of P11274 - P00519 specific RNA by RT-PCR has been shown to predict relapse when positive 6 months after allogeneic stem cell transplantation ( P09683 ) for chronic myelogenous leukemia ( CML ) . In the present study , the focus was on evaluation of residual disease during the first weeks following P09683 . In this study , 177 blood or marrow samples were obtained from 33 patients who received allogeneic ( 20 patients ) or autologous ( 13 patients ) P09683 on day 0 , day 30 and every 3 months for 1 year . T-cell depletion ( P24386 ) was performed in 4 cases . On day 0 ( day of graft infusion ) , 10/30 evaluable patients had negative RT-PCR ( 33 % ) regardless of pretransplant characteristics . On day 30 , 14/18 patients ( 77 % ) from the allogeneic group had negative RT-PCR versus 0 % in the autologous group . 2/4 patients who received P24386 allogeneic grafts had day 30-positive PCR . Five patients in the allogeneic group had at least one positive RT-PCR sample between day 30 and day 90 : 3 of them subsequently relapsed suggesting possible correlation between early positivity and relapse . Our results show that disappearance of MRD can be achieved within 3 months after transplantation in the majority of patients treated with allogeneic but not after autologous P09683 . This suggests that the GVL effect might be operational early during the first weeks following transplantation .

MH_train_1115

MH_train_1115

MH_train_1115

interacts_with DB00945?

multiple_choice

Selective inhibition of prostaglandin endoperoxide synthase-1 ( cyclooxygenase-1 ) by valerylsalicylic acid . DB00945 causes a time-dependent inhibition of prostaglandin endoperoxide H synthases ( PGHS ) -1 and -2 by acetylating active site serines present in both isozymes . In the case of P23219 , aspirin acetylation blocks cyclooxygenase activity , apparently by preventing arachidonate binding to the cyclooxygenase active site . With P35354 , acetylation does not block substrate binding but rather alters the enzyme in such a way that the acetylated form of P35354 produces 15R-hydroxyeicosatetraenoic acid ( 15R-HETE ) instead of the usual prostaglandin endoperoxide product . Based on these differences between P23219 and P35354 , we reasoned that a salicylate ester containing an acyl group somewhat larger than the acetyl group of aspirin might be a selective inhibitor of P35354 . Accordingly , we prepared and tested eight different acyl salicylates as inhibitors of human ( h ) P23219 and -2 expressed transiently in cos-1 cells . Valeryl(pentanoyl)salicylate ( VSA ) was the only compound in this series which showed isozyme selectivity , and , surprisingly , VSA inhibited hPGHS-1 much more effectively than hPGHS-2 . Inhibition of hPGHS-1 by VSA was time-dependent . VSA also inhibited ovine P23219 but did not inhibit the S530A mutant of ovine P23219 . This latter mutant , which lacks the active site serine hydroxyl group , is also refractory to inhibition by acetylsalicylate . Thus , we conclude that VSA acylates the active site serine of P23219 . VSA inhibited prostanoid synthesis by serum-starved murine NIH 3T3 cells which express only P23219 ; in contrast , VSA caused only partial inhibition of prostanoid synthesis by serum-stimulated 3T3 cells which express both PGHS isozymes . Our results establish that VSA can be used as a reasonably selective inhibitor of P23219 . Potential cardiovascular effects of P35354 selective nonsteroidal antiinflammatory drugs . The newly developed nonsteroidal antiinflammatory drugs ( NSAIDs ) that selectively inhibit cyclooxygenase-2 ( P35354 ) , are effective against pain and inflammation and appear to have less gastrointestinal toxicity than conventional NSAIDs . Their P35354 selectivity , however , has raised concerns regarding their cardiovascular safety , since they do not inhibit P23219 , the isoform of the enzyme that is active in thrombosis and vasoconstriction . At this point there is no conclusive evidence that P35354 inhibitors cause ischemic vascular events , because retrospective post hoc analyses conflict one another , and no specific randomized trials have yet been done . Renal effects , edema and hypertension appear to be similar between conventional NSAIDs and P35354 -selective inhibitors . DB00945 is still required for patients with cardiovascular risk who are prescribed a P35354 -selective inhibitor . Participation of tyrosine phosphorylation in cytoskeletal reorganization , alpha(IIb)beta(3) integrin receptor activation , and aspirin-insensitive mechanisms of thrombin-stimulated human platelets . BACKGROUND : DB09222 binding to the active conformation of the alpha(IIb)beta(3) integrin receptor ( glycoprotein IIb/IIIa ) and cytoskeletal reorganization are important events in platelet function . Tyrosine phosphorylation of platelet proteins plays an essential role in platelet signal transduction pathways . We studied the participation of tyrosine kinases on these aspects of platelet reactivity and their importance in cyclooxygenase ( P36551 ) -1-independent mechanisms in thrombin-stimulated human platelets . METHODS AND RESULTS : Using washed platelets from normal donors and tyrphostin-A47 and aspirin as tyrosine kinase and P23219 inhibitors , respectively , we found that tyrphostin-A47 downregulated ( 1 ) the thrombin-activated conformational change of alpha(IIb)beta(3) , ( 2 ) actin polymerization and cytoskeletal reorganization , and ( 3 ) the quantity of tyrosine-phospho-rylated proteins associated with the reorganized cytoskeleton . The latter are important components of multimolecular signaling complexes . Concomitantly , platelet aggregation and secretion were significantly reduced . DB00945 did not affect receptor activation or tyrosine phosphorylation but did decrease the initial ( 30-second ) burst of actin polymerization . Importantly , aspirin significantly amplified the inhibitory effect of tyrphostin-A47 on all aspects of platelet reactivity that we evaluated . CONCLUSIONS : Tyrosine protein phosphorylation is a regulatory control system of the inside-out mechanism of alpha(IIb)beta(3) activation and cytoskeletal assembly in thrombin-stimulated human platelets . Inhibition of these aspects of platelet function with tyrphostin-A47 is amplified when platelets are treated with aspirin . Therefore , tyrosine phosphorylation is a major component of early signaling events and of P23219 -independent mechanisms of thrombin-induced platelet reactivity . The study results may indicate a novel target for therapeutic intervention . Proapoptotic and antiproliferative potential of selective cyclooxygenase-2 inhibitors in human liver tumor cells . Recent studies have shown increased levels of cyclooxygenase-2 ( P35354 ) in a variety of human malignancies , including hepatocellular carcinoma ( HCC ) , but so far it is unknown whether P35354 contributes to the malignant growth and whether inhibition of P35354 function modifies the malignant potential of liver tumors . P23219 and P35354 expression was determined in 4 liver tumor cell lines ( Hep 3B , HuH-7 , Hep G2 , Sk-hep1 ) by Northern hybridization and Western immunoblot . The functional effects of the nonselective inhibitor sulindac sulfide and the P35354 selective inhibitors SC-58635 and meloxicam were examined by 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide ( MTT ) -assays and BrdU uptake , morphology , and TUNEL analysis of apoptosis . Apoptosis regulating proteins were analyzed by Western immunoblot . P23219 and P35354 expression was demonstrable in all tested liver tumor cell lines . Sulindac sulfide ( 50 to 400 micromol/L ) , SC-58635 ( 6,25 to 400 micromol/L ) , and meloxicam ( 6.25 to 400 micromol/L ) led to a significant time- and dose-dependent reduction of cell numbers of up to 80 % ( P < .05 ) . At equimolar concentrations the effect was more pronounced when P35354 was selectively blocked . P35354 inhibition induced apoptosis and reduced tumor cell proliferation . Apoptosis after P35354 inhibition with SC-58635 ( 50 micromol/L ) was independent of BCL-2 , Q07812 , and the phosphorylation status of AKT/ P31749 and Q92934 , but correlated with activation of caspase-9 , caspase-3 , and caspase-6 . In conclusion , selective inhibition of P35354 leads to a marked growth inhibition of human liver tumor cells , based on the induction of apoptosis and inhibition of proliferation and , thus , may offer therapeutic and preventive potential in human hepatocarcinogenesis . 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 . Nuclear factor kappaB ( NF-kappaB ) pathway as a therapeutic target in rheumatoid arthritis . Rheumatoid arthritis ( RA ) is a chronic inflammatory disease characterized by persistent joint swelling and progressive destruction of cartilage and bone . Current RA treatments are largely empirical in origin and their precise mechanism of action is uncertain . Increasing evidence shows that chronic inflammatory diseases such as RA are caused by prolonged production of proinflammatory cytokines including tumor necrosis factor ( P01375 ) and interleukin 1 ( IL-1 ) . The nuclear factor kappaB ( NF-kappaB ) plays an essential role in transcriptional activation of P01375 and IL-1 . NF-kappaB is induced by many stimuli including P01375 and IL-1 , forming a positive regulatory cycle that may amplify and maintain RA disease process . NF-kappaB and enzymes involved in its activation can be a target for anti-inflammatory treatment . DB00945 and sodium salicylate inhibit activation of NF-KB by blocking O15111 , a key enzyme in NF-kappaB activation . Glucocorticoids suppress expression of inflammatory genes by binding glucocorticoid receptor with NF-kappaB , and increasing expression of inhibitory protein of NF-kappaB , P25963 . Sulfasalazine and gold compounds also inhibit NF-kappaB activation . Continuing advances in our understanding of action mechanism of antirheumatic agents will benefit the future development of RA regimens with greater efficacy and less toxicity . 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 . 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 . Analysis of P25054 types involved in P01730 tolerance and regulatory T cell generation using reaggregated thymic organ cultures . Tolerance to self-Ags is generated in the thymus . Both epithelial and hematopoietic thymic stromal cells play an active and essential role in this process . However , the role of each of the various stromal cell types remains unresolved . To our knowledge , we describe the first comparative analysis of several types of thymic hematopoietic stromal cells ( THSCs ) for their ability to induce P01730 tolerance to self , in parallel with the thymic epithelium . The THSCs -- two types of conventional dendritic cells ( cDCs ) , plasmacytoid dendritic cells , macrophages ( MΦs ) , B lymphocytes , and eosinophils -- were first characterized and quantified in adult mouse thymus . They were then examined in reaggregated thymic organ cultures containing mixtures of monoclonal and polyclonal thymocytes . This thymocyte mixture allows for the analysis of Ag-specific events while avoiding the extreme skewing frequently seen in purely monoclonal systems . Our data indicate that thymic epithelium alone is capable of promoting self-tolerance by eliminating autoreactive P01730 single-positive thymocytes and by supporting regulatory T cell ( Treg ) development . We also show that both non-Treg P01730 single-positive thymocytes and Tregs are efficiently deleted by the two populations of cDCs present in the thymus , as well as to a lesser extent by MΦs . Plasmacytoid dendritic cells , B lymphocytes , and eosinophils were not able to do so . Finally , cDCs were also the most efficient THSCs at supporting Treg development in the thymus , suggesting that although they may share some characteristics required for negative selection with MΦs , they do not share those required for the support of Treg development , making cDCs a unique cell subset in the thymus . 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 lipoxygenase-cyclooxygenase inhibitor licofelone prevents thromboxane A2-mediated cardiovascular derangement triggered by the inflammatory peptide fMLP in the rabbit . DB04725 is an analogue of arachidonic acid that inhibits P09917 ( P28300 ) , cyclooxygenase ( P36551 ) -1 and P35354 . We investigated the effects of licofelone on cardiovascular derangements and production of thromboxane (Tx)A(2) induced by the inflammatory agonist n-formyl-methionyl-leucyl-phenylalanine ( fMLP ) in the rabbit , in comparison with those of aspirin or rofecoxib , inhibitors of P23219 and P35354 , respectively . In control rabbits , injection of fMLP ( 30 nmol/kg ) in the jugular vein evokes ischemic electrocardiographic ( ECG ) changes in the first 1-5 min , i.e. a profound depression of the ST segment and inversion of the T wave . Simultaneously , fMLP induces bradycardia and hypotension and increases TxB(2) blood levels . All changes are transient . DB04725 ( 60 mg/kg/5 days , p.os ) prevented fMLP-induced ECG ischemic changes in all treated animals , reverted bradycardia and hypotension , and significantly reduced TxB(2) . DB00945 ( 10 mg/kg/5 days , p.os ) prevented ischemic ECG alterations in 2 out of 5 treated animals and did not modify either bradycardia or hypotension . One rabbit died two min after fMLP . In 2 rabbits , aspirin reduced TxB(2) levels by more than 80 % respect to mean control values ; the remaining two rabbits produced an amount of TxB(2) similar to controls . These two rabbits also showed ischemic ECG changes . DB00533 ( 10 mg/kg/5 days , p.os ) did not prevent fMLP-induced ischemic ECG alteration , bradycardia and hypotension , and did not significantly modify the increase of TxB(2) . These results indicate that the capacity of licofelone to efficiently suppress TxA(2) production , is responsible for the protection from the cardiovascular derangement triggered by an inflammatory stimulus . Functional and biochemical evaluation of platelet aspirin resistance after coronary artery bypass surgery . BACKGROUND : DB00945 inhibits platelet activation and reduces atherothrombotic complications in patients at risk of myocardial infarction and stroke . However , a sufficient inhibition of platelet function by aspirin is not always achieved . The causes of this aspirin resistance are unknown . METHODS AND RESULTS : Patients undergoing coronary artery bypass grafting ( CABG ) have a high incidence of aspirin resistance . To evaluate functional and biochemical responses to aspirin , platelet-rich plasma was obtained before and at days 1 , 5 , and 10 after CABG . Thromboxane formation , aggregation , and alpha-granule secretion were effectively inhibited by 30 or 100 micromol/L aspirin in vitro before CABG , but this inhibition was prevented or attenuated after CABG . Whereas the inhibition of thromboxane formation and aggregation by aspirin in vitro partly recovered at day 10 after CABG , oral aspirin ( 100 mg/d ) remained ineffective . The inducible isoform of cyclooxygenase in platelets , P35354 , has been suggested to confer aspirin resistance . In fact , immunoreactive P35354 was increased 16-fold in platelets at day 5 after CABG , but the P35354 selective inhibitor celecoxib did not alter aspirin-resistant thromboxane formation . By contrast , the combined inhibitor of thromboxane synthase and thromboxane receptor antagonist terbogrel equally prevented thromboxane formation of platelets obtained before ( control ) and after CABG . CONCLUSIONS : Platelet aspirin resistance involves an impairment of both in vivo and in vitro inhibition of platelet functions and is probably due to a disturbed inhibition of platelet P23219 by aspirin . New direct and indirect methods for the detection of cyclooxygenase 1 acetylation by aspirin ; the lack of aspirin resistance among healthy individuals . BACKGROUND : DB00945 is widely used in the prevention of acute atherothrombotic complications . It acetylates Ser529 residue in cyclooxygenase-1 ( P23219 ) and prevents thromboxane A2 ( TXA2 ) formation from arachidonic acid ( AA ) in platelets . Laboratory methods used for the detection of aspirin effect provide inconsistent results . METHODS : Two new methods were developed for the direct and indirect detection of P23219 acetylation by aspirin in 108 healthy volunteers treated daily with 100mg enteric-coated aspirin for 7days . Monoclonal antibodies were raised against acetylated and non-acetylated nonapeptides corresponding to the amino acid sequence of human P23219 525-533 residues . Using Western blotting technique the antibodies clearly distinguished between acetylated and non-acetylated P23219 in platelet lysate . The second method measures AA-induced TXB2 production of platelets in diluted platelet rich plasma . RESULTS : No acetylated P23219 was detected in platelets before aspirin treatment . At the same time antibodies raised against non-acetylated peptide gave intense reaction with P23219 on the Western blot . In contrast , after 7days of aspirin treatment , with a single exception , only acetylated P23219 could be detected in the platelet lysate . The non-responding volunteer showed full response to aspirin after controlled drug intake . In parallel experiments aspirin treatment for 7days practically completely inhibited AA-induced TXB2 production by platelets . CONCLUSIONS : Chemical ( " true " ) aspirin resistance , if it exists , must be a rarity among healthy individuals . The new methods could be used for detecting the acetylation of P23219 by aspirin in patients on preventive aspirin therapy and for evaluating methods routinely used for such purpose . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . Apple polyphenol extracts prevent aspirin-induced damage to the rat gastric mucosa . DB00945 causes gastroduodenal ulcers and complications . Food bioactive compounds could exert beneficial effects in the gastrointestinal tract . We evaluated whether apple polyphenol extract ( APE ) reduced aspirin-induced injury to the rat gastric mucosa . Rats were treated with APE ( 10(-4) m catechin equivalent ) before oral aspirin ( 200 mg/kg ) . Cyclo-oxygenase-2 ( P35354 ) , transforming growth factor-alpha ( TGF alpha ) and heparin-binding epidermal-growth-factor-like growth factor ( HB- P01133 ) mRNA and protein expression were assessed by RT-PCR and Western blot analysis , respectively ; malondialdehyde ( MDA ) was determined by HPLC ; gastric secretion was evaluated in pylorus-ligated rats . APE decreased acute and chronic aspirin injury both macroscopically and microscopically ( approximately 50 % decrease in lesion score ; P < 0.05 ) . DB00945 up-regulated mRNA and protein expression of P35354 and HB- P01133 , but not of TGF alpha ; APE reduced aspirin-induced mRNA and protein over-expression of P35354 and HB- P01133 ; aspirin significantly increased gastric MDA and this effect was counteracted by APE pre-treatment . APE did not significantly affect gastric acid secretion . In conclusion , APE reduces aspirin-induced gastric injury independently of acid inhibition . We speculate that APE might be of therapeutic use in the prophylaxis of aspirin-related gastropathy . IL-1 does not reverse the anti-proliferative effect of aspirin in breast cancer cells . OBJECTIVES : Interleukin-1 ( IL-1 ) is a multifunctional proinflammatory cytokine . There have been studies suggesting a role in affecting growth and invasiveness of malignant breast cells by either blocking or stimulating growth of cultured MCF-7 breast cancer cells . This effect may be mediated by induction of P35354 . DB00945 is an inhibitor of P35354 and has been implicated , with other non-steroidal anti-inflammatory drugs ( NSAIDS ) in prevention and treatment of breast cancer . In this study the in vitro effects of IL-1 and aspirin on growth of MCF-7 human breast cancer cells was examined . METHODS : MCF-7 cells were treated with various concentrations of IL-1 and aspirin alone and in combination . Cell growth was assessed by cell number measurement . RESULTS : DB00945 significantly decreased growth rate in a dose-dependent manner , alone and as a combined treatment with IL-1 with a maximum reduction in growth rate at 300 mg/ml ( P < 0.05 ) . Treatment with IL-1 alone showed no significant effect on growth rate of MCF-7 cells ( P > 0.05 ) . CONCLUSION : This study confirms that aspirin suppresses the proliferation rate of MCF-7 cells both as a single agent and in combination with IL-1 . It also suggests that IL-1 alone does not stimulate or inhibit growth of MCF-7 cells . Getting to the heart of P35354 inhibition . DB00945 protects against heart disease , while P35354 Inhibitors appear to injure the heart . The studies by [ this Issue of Cell Metabolism ] ) shed light on two distinct prostaglandin receptors contributing to these conflicting cardiovascular effects . 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 . 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 . 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 . DB00945 attenuates cytomegalovirus infectivity and gene expression mediated by cyclooxygenase-2 in coronary artery smooth muscle cells . Human cytomegalovirus ( CMV ) infection of smooth muscle cells generates reactive oxygen species ( ROS ) and thereby activates nuclear factor kappaB ( NFkappaB ) , which causes expression of viral and cellular genes involved in immune and inflammatory responses . These changes could account for the mounting evidence suggesting that CMV may contribute causally to restenosis and atherosclerosis . We found that CMV induces ROS , at least partly , through a cyclooxygenase-2 ( P35354 ) -dependent pathway . Moreover , the viral immediate-early ( IE ) gene products , IE72 and IE84 , have the capacity to transactivate the P35354 promoter . DB00945 and indomethacin , both cyclooxygenase inhibitors as well as direct ROS scavengers , reduce CMV-induced ROS , probably through both of these activities . Sodium salicylate also has antiviral effects as the result of its potent antioxidant properties . Furthermore , by reducing ROS , aspirin and sodium salicylate inhibit CMV-induced NFkappaB activation , the ability of IE72 to transactivate its promoter , CMV IE gene expression after infection of SMCs , and CMV replication in SMCs . This is the first time aspirin has been shown to have antiviral effects . Thus , it is possible that aspirin has previously unrecognized therapeutic effects in various clinical situations , such as in viral infections ( when used as an antipyretic agent ) and in atherosclerosis ( when used as an antiplatelet agent ) . [ DB00945 and prevention of colorectal carcinomas ] . Observational and some randomized clinical trials suggest that aspirin protects from occurrence and progression of colorectal neoplasias ( adenomas , carcinomas ) . However , there are still open questions , regarding the benefit/risk ratio ( bleedings ) as well as dosage and duration of treatment during the probably long-term medication , before stringent recommendations regarding clinical use of aspirin can be made . Specifically , there is currently no generally accepted mode of action or molecular target of aspirin , though a relationship to tumor-associated enhanced DB00917 levels in the affected mucosa is likely . Regular daily intake of aspirin in antiplatelet doses of 100 mg appears to be sufficient in responding persons . If this is confirmed in prospective randomized trials that are currently underway , this might add to the prophylactic use of aspirin and would suggest a pharmacological relationship to inhibition of P23219 mediated prostaglandin/thromboxane biosynthesis as a common primary target for both cardiocoronary and antineoplastic prophylaxis . Prophylactic aspirin use might then add to an undoubtedly important healthy lifestyle including appropriate diet . Platelet thromboxane ( 11-dehydro-Thromboxane B2 ) and aspirin response in patients with diabetes and coronary artery disease . DB00945 ( ASA ) irreversibly inhibits platelet cyclooxygenase-1 ( P23219 ) leading to decreased thromboxane-mediated platelet activation . The effect of ASA ingestion on thromboxane generation was evaluated in patients with diabetes ( DM ) and cardiovascular disease . Thromboxane inhibition was assessed by measuring the urinary excretion of 11-dehydro-thromboxane B2 ( 11dhTxB2 ) , a stable metabolite of thromboxane A2 . The mean baseline urinary 11dhTxB2 of DM was 69.6 % higher than healthy controls ( P = 0.024 ) : female subjects ( DM and controls ) had 50.9 % higher baseline 11dhTxB2 than males ( P = 0.0004 ) , while age or disease duration had no influence . Daily ASA ingestion inhibited urinary 11dhTxB2 in both DM ( 71.7 % ) and controls ( 75.1 % , P < 0.0001 ) . Using a pre-established cut-off of 1500 pg/mg of urinary 11dhTxB2 , there were twice as many ASA poor responders ( ASA " resistant " ) in DM than in controls ( 14.8 % and 8.4 % , respectively ) . The rate of ASA poor responders in two populations of acute coronary syndrome ( ACS ) patients was 28.6 and 28.7 % , in spite of a significant ( 81.6 % ) inhibition of urinary 11dhTxB2 ( P < 0.0001 ) . Both baseline 11dhTxB2 levels and rate of poor ASA responders were significantly higher in DM and ACS compared to controls . Underlying systemic oxidative inflammation may maintain platelet function in atherosclerotic cardiovascular disease irrespective of P23219 pathway inhibition and/or increase systemic generation of thromboxane from non-platelet sources . 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 . DB00945 , but not NO-releasing aspirin ( O43763 -4016 ) , interacts with selective P35354 inhibitors to aggravate gastric damage and inflammation . Aceylation of cyclooxygenase ( P36551 ) -2 by aspirin can trigger the formation of 15(R)-epilipoxin A4 , or aspirin-triggered lipoxin ( ATL ) . ATL exerts protective effects in the stomach . Selective P35354 inhibitors block ATL synthesis and exacerbate aspirin-induced gastric damage . DB00435 -releasing aspirins , including O43763 -4016 , have antiplatelet effects similar to aspirin but do not cause gastric damage . In the present study , we examined whether or not O43763 -4016 triggers ATL synthesis and/or upregulates gastric P35354 expression and the effects of coadministration of O43763 -4016 with a selective P35354 inhibitor on gastric mucosal injury and inflammation . Rats were given aspirin or O43763 -4016 orally and either vehicle or a selective P35354 inhibitor ( celecoxib ) intraperitoneally . Gastric damage was blindly scored , and granulocyte infiltration into gastric tissue was monitored through measurement of myeloperoxidase activity . Gastric PG and ATL synthesis was measured as was P35354 expression . Whereas celecoxib inhibited gastric ATL synthesis and increased the severity of aspirin-induced gastric damage and inflammation , coadministration of celecoxib and O43763 -4016 did not result in damage or inflammation . O43763 -4016 did not upregulate gastric P35354 expression nor did it trigger ATL synthesis ( in contrast to aspirin ) . Daily administration of aspirin for 5 days resulted in significantly less gastric damage than that seen with a single dose , as well as augmented ATL synthesis . Celecoxib reversed this effect . In contrast , repeated administration of O43763 -4016 failed to cause gastric damage , whether given alone or with celecoxib . These studies support the notion that O43763 -4016 may be an attractive alternative to aspirin for indications such as cardioprotection , including in individuals also taking selective P35354 inhibitors . Expression of cyclooxygenase-2 ( P35354 ) in tumour and stroma compartments in cervical cancer : clinical implications . This study aims at investigating the relationship between cyclooxygenase-2 expression in tumour vs stroma inflammatory compartment and its possible clinical role . The study included 99 stage IB-IV cervical cancer patients : immunostaining of tumour tissue sections was performed with rabbit antiserum against cyclooxygenase-2 . CD3 , P01730 , CD8 , CD25 , Mast Cell Tryptase monoclonal antibodies were used to characterise stroma inflammatory cells in nine cervical tumours . An inverse relation was found between cyclooxygenase-2 levels ( cyclooxygenase-2 IDV ) of tumour vs stroma compartment ( r=-0.44 , P < 0.0001 ) . The percentage of cases showing high tumour/stromal cyclooxygenase-2 IDV ratio was significantly higher in patients who did not respond to treatment ( 93.3 % ) with respect to patients with partial ( 60.5 % ) , and complete ( 43.7 % ) response ( P= 0.009 ) . Cases with a high tumour/stroma cyclooxygenase-2 IDV ratio had a shorter overall survival rate than cases with a low tumour/stroma cyclooxygenase-2 IDV ( P < 0.0001 ) . In the multivariate analysis advanced stage and the status of tumour/stroma cyclooxygenase-2 IDV ratio retained an independent negative prognostic role . The proportion of CD3(+) , P01730 (+) , and CD25(+) cells was significantly lower in tumours with high tumour/stroma cyclooxygenase-2 IDV ratio , while a higher percentage of mast cells was detected in tumours showing high tumour/stroma cyclooxygenase-2 IDV ratio . Our study showed the usefulness of assessing cyclooxygenase-2 status both in tumour and stroma compartment in order to identify cervical cancer patients endowed with a very poor chance of response to neoadjuvant therapy and unfavourable prognosis . 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 . 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 . Endogenous concentrations of ouabain act as a cofactor to stimulate fluid secretion and cyst growth of in vitro ADPKD models via DB02527 and P00533 -Src-MEK pathways . In autosomal-dominant polycystic kidney disease ( ADPKD ) , renal cysts develop by aberrant epithelial cell proliferation and transepithelial fluid secretion . We previously showed that ouabain increases proliferation of cultured human ADPKD cells via stimulation of the P01133 receptor ( P00533 ) -Src-MEK/ P29323 signaling pathway . We examined whether ouabain affects fluid secretion and in vitro cyst growth of human ADPKD cell monolayers , ADPKD cell microcysts cultured in a three-dimensional collagen matrix , and metanephric organ cultures from Pkd1(m1Bei) mice . Physiological concentrations of ouabain alone did not affect net transepithelial basal-to-apical fluid transport in ADPKD monolayers or growth of cultured ADPKD microcysts . In contrast , in the presence of forskolin or 8-bromo- DB02527 , ouabain significantly enhanced ADPKD fluid secretion and microcyst expansion . Ouabain exerted this effect by enhancing DB02527 -dependent Cl(-) secretion via the P13569 . Similarly , ouabain accelerated DB02527 -dependent cyst enlargement in Pkd1(m1Bei) mice metanephroi , with a more prominent response in homozygous than heterozygous mice . Ouabain had no effect on fluid secretion and cystogenesis of normal human kidney cells and caused only slight cystic dilations in wild-type mouse kidneys . The effects of ouabain in ADPKD cells and Pkd1(m1Bei) metanephroi were prevented by inhibitors of P00533 ( AG1478 ) , Src ( Q99463 ) , and MEK ( U0126 ) . Together , our results show that ouabain , used in physiological concentrations , has synergistic effects on DB02527 -mediated fluid secretion and cyst growth , via activation of the P00533 -Src-MEK pathway . These data provide important evidence for the role of ouabain as an endogenous hormone that exacerbates ADPKD cyst progression . Sites of action for future therapy : an adenosine-dependent mechanism by which aspirin retains its antiinflammatory activity in cyclooxygenase-2 and NFkappaB knockout mice . The antiinflammatory action of aspirin is generally attributed to inhibition of cyclooxygenases 1 and 2 , but additional mechanisms are at work . These include inhibition of NFkappaB translocation to the nucleus and the capacity of aspirin to promote accumulation of adenosine , a potent antiinflammatory autocoid . We tested these hypotheses in the murine air pouch model of acute inflammation in wild type mice and in cyclooxygenase 2 or NFkappaB knockouts . The antiinflammatory effects of aspirin , sodium salicylate and indomethacin did not correlate with inhibition of cyclooxygenase in either group . Indeed , aspirin retained its antiinflammatory properties even in P35354 knockouts . Similarly , aspirin was no less antiinflammatory in mice rendered deficient for NFkappaB ( Q14511 ) than in wild type controls . In contrast , dexamethasone lost its antiinflammatory capacity in NFkappaB knockouts . DB00945 and sodium salicylate dramatically increased concentrations of adenosine in exudates , a property shared with methotrexate and sulfasalazine . Removal of adenosine by adenosine deaminase or specific antagonism of adenosine at A(2)receptors completely reversed the antiinflammatory effects of aspirin and sodium salicylate , but not those of dexamethasone . This adenosine-dependent , antiinflammatory effect of aspirin points to another target of drug development . P01375 -alpha and X-linked adrenoleukodystrophy . The two most common forms of X-linked adrenoleukodystrophy ( X- P33897 ) , the childhood cerebral form ( CCER ) and the adult form , adrenomyeloneuropathy ( Q9BXJ7 ) , arise from the same mutations in the X- P33897 gene at Xq28 . These two forms are distinguished by the degree of cerebral inflammation . Segregation analysis suggests that an autosomal modifying gene may be a major determinant of phenotype in X- P33897 . Thus , a modifying gene could be involved in initiating or promoting the inflammatory response . In this study we detected a difference in tumor necrosis factor-alpha ( P01375 ) bioactivity , but not P01375 protein levels , in serum from some advanced CCER patients . Early-stage CCER patients and Q9BXJ7 patients were in the normal range . Allelic differences in P01375 or levels of soluble P01375 receptor did not account for bioactivity differences or phenotypic heterogeneity in X- P33897 . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . DB00945 causes rapid up-regulation of cyclo-oxygenase-2 expression in the stomach of rats . BACKGROUND : Cyclo-oxygenase-1 ( P23219 ) is believed to produce prostaglandins vital to mucosal defence , whereas cyclo-oxygenase-2 ( P35354 ) is induced at sites of inflammation . Little is known about the regulation of P35354 in the stomach , particularly during the period following mucosal injury . In this study , we examined P23219 and P35354 expression shortly after administration of NSAIDs or ethanol . METHODS : Fasted rats were given aspirin , salicylate , indomethacin or ethanol ( 20 % or 40 % ) orally . Three hours later the stomach was excised , the severity of damage scored and samples taken for RT-PCR of P23219 and P35354 mRNA and immunohistochemistry . DB00435 synthase mRNA ( P35228 and P29474 ) and activity were also measured . RESULTS : DB00945 , indomethacin and the higher concentration of ethanol produced widespread mucosal damage , whereas salicylate and 20 % ethanol caused only superficial epithelial damage . DB00945 caused a significant increase in P35354 mRNA expression and a marked increase in P35354 immunoreactivity , particularly in the superficial mucosa . Expression of P23219 ( mRNA and protein ) was unaffected by aspirin , as were NOS mRNA expression and enzyme activity . Pre-treatment with prostaglandin E2 prevented the induction of P35354 by aspirin . Salicylate and indomethacin caused modest increases in P35354 immunoreactivity but no change in P35354 mRNA . Neither concentration of ethanol affected P35354 mRNA or protein expression , suggesting that this was a specific response to the aspirin , rather than to injury . CONCLUSIONS : These results demonstrate a rapid up-regulation of P35354 expression in response to aspirin , possibly representing a compensatory response to inhibition of gastric prostaglandin synthesis . The effect of aspirin and nonsteroidal anti-inflammatory drugs on prostaglandins . Cyclic prostanoids play important physiologic roles in inflammation and maintaining normal function of several organ systems . Prostaglandin production requires the conversion of arachidonate to the intermediate prostaglandin H2 , by the 2-step cyclo-oxygenation and peroxidation catalyzed by the enzyme cyclo-oxygenase ( also called prostaglandin H synthase ) . DB00945 and nonsteroidal anti-inflammatory drugs ( NSAIDs ) block the production of cyclic prostanoids by binding in different ways to this enzyme and blocking the active site . This results in decreased inflammation , but it can also produce side effects in the gastrointestinal tract , kidney , and platelets . Recent data demonstrate that there are 2 isoforms of the cyclo-oxygenase enzyme , called P23219 and P35354 . These isoforms are similar in size , substrate specificity , and kinetics , but vary in their expression and distribution . Normal physiologic functions appear to be maintained by P23219 , while P35354 appears to mediate the inflammatory response . Nonsteroidal drugs with selective inhibitory activity on the P35354 isoform should theoretically decrease inflammation while maintaining normal physiologic prostaglandin levels . Current NSAIDs are not selective enough to confirm this , but newer , more selective inhibitors of P35354 may answer this important question . P35354 : Where are we in 2003 ? - Specific cyclooxygenase-2 inhibitors and aspirin-exacerbated respiratory disease . The use of analgesic anti-inflammatory agents in patients with asthma is clinically challenging because of the prevalence ( 10-20 % ) of aspirin hypersensitivity . DB00945 -exacerbated respiratory disease ( AERD ) , or aspirin-induced asthma , is characterized by asthma and rhinitis triggered by the ingestion of aspirin and non-steroidal anti-inflammatory drugs . AERD is associated with upper and lower respiratory-tract mucosal inflammation , progressive sinusitis , nasal polyposis , and asthma regardless of whether patients avoid triggering drugs . The mechanism underlying the propensity of aspirin and non-steroidal anti-inflammatory drugs to cause this reaction is thought to involve inhibition of the synthesis of protective prostaglandins ( PGs ) , resulting in an increase in the synthesis of cysteinyl leukotrienes by eosinophils and mast cells . Clinical data suggest that protective PGs are derived from cyclooxygenase ( P36551 ) -1 because studies have now confirmed that drugs specifically inhibiting P35354 are not cross-reactive with aspirin in patients with AERD . Non-small cell lung cancer cycloxygenase activity and proliferation are inhibited by non-steroidal antiinflammatory drugs . The effects of non-steroidal antiinflammatory drugs ( NSAIDs ) on non-small cell lung cancer ( NSCLC ) were investigated . Arachidonic acid ( AA ) was metabolized to prostaglandin E2 ( DB00917 ) in NSCLC cells . NSAIDs such as aspirin or indomethacin reduced DB00917 levels in NCI-H157 and H1264 cells , and the decrease caused by DB00917 was reversed by epidermal growth factor ( P01133 ) . By RT-PCR , both cyclooxygenase ( P36551 ) -1 and P35354 mRNAs are detected in NCI-H157 and H1264 cells . By Northern analysis , P35354 mRNA was induced by P01133 and phorbol ester . By immunocytochemistry , P23219 and P35354 enzymes were localized to NSCLC tumors . DB00945 , indomethacin and ibuprofen decreased NSCLC growth in vitro . DB00945 and indomethacin inhibited proliferation of NSCLC xenografts in nude mice . These data suggest that P36551 enzymes may be important regulatory components of NSCLC . Intracellular signaling as a potential target for antiplatelet therapy . Three classes of inhibitors of platelet aggregation have demonstrated substantial clinical benfits . DB00945 acts by irreversibly inhibiting P23219 and therefore blocking the synthesis of proaggregatory thromboxane A ( 2 ) ( TxA(2) ) . The indirect acting ( ticlopidine , clopidogrel , prasugrel ) and the direct acting ( ticagrelor ) antagonists of P2Y(12) block the thrombus stabilizing activity of ADP . Parenteral GP IIb-IIIa inhibitors directly block platelet-platelet interactions . Despite well-established benefits , all antiplatelet agents have important limitations : increased bleeding and gastrointestinal toxicities ( aspirin ) , high incidence of thrombotic thrombocytopenic purpura ( ticlopidine ) , potentially nonresponders ( clopidogrel ) , severe bleeding ( prasugrel , GP IIb-IIIa antagonists ) and " complicated " relationships with aspirin ticagrelor ) . In this chapter , we present the genetic and pharmacological evidence that supports the development and expectations associated with novel antiplatelet strategies directed at intrasignaling pathways . Reed-Sternberg cell-derived lymphotoxin-α activates endothelial cells to enhance T-cell recruitment in classical Hodgkin lymphoma . It is known that cells within the inflammatory background in classical Hodgkin lymphoma ( cHL ) provide signals essential for the continual survival of the neoplastic Hodgkin and Reed-Sternberg ( HRS ) cells . However , the mechanisms underlying the recruitment of this inflammatory infiltrate into the involved lymph nodes are less well understood . In this study , we show in vitro that HRS cells secrete lymphotoxin-α ( LTα ) which acts on endothelial cells to upregulate the expression of adhesion molecules that are important for T cell recruitment . LTα also enhances the expression of hyaluronan which preferentially contributes to the recruitment of P01730 (+) CD45RA(+) naïve T cells under in vitro defined flow conditions . Enhanced expression of LTα in HRS cells and tissue stroma ; and hyaluronan on endothelial cells are readily detected in involved lymph nodes from cHL patients . Our study also shows that although NF-κB and AP-1 are involved , the cyclooxygenase ( P36551 ) pathway is the dominant regulator of LTα production in HRS cells . Using pharmacological inhibitors , our data suggest that activity of P23219 , but not of P35354 , directly regulates the expression of nuclear c-Fos in HRS cells . Our findings suggest that HRS cell-derived LTα is an important mediator that contributes to T cell recruitment into lesional lymph nodes in cHL . The comparative study of acetyl-11-keto-beta-boswellic acid ( AKBA ) and aspirin in the prevention of intestinal adenomatous polyposis in P25054 (Min/+) mice . Acetyl-11-keto-beta-BA ( AKBA ) , a component of the gum resin of Boswellia serrata , has been recognized as a promising agent for the prevention of intestinal tumorigenesis . DB00945 , a non-steroidal anti-inflammatory drug ( NSAID ) , has also been considered to have the activity against intestinal tumorigenesis . However , the prevention of colonic cancer is insufficient and no definitive recommendation has been made for clinic use . Herein , we compared the efficacy of AKBA with that of aspirin in an adenomatous polyposis coli intestinal neoplasia consecutive weeks . Mice were sacrificed by anesthetizing . The whole intestine was removed from each mouse . The number , size and histopathology of intestinal adenomatous polyps were examined under microscopy . The adenomatous polyps were removed for further analysis by the assays of western blotting and immunohistochemical staining . AKBA significantly prevented the formation of intestinal adenomatous polyps without toxicity to mice . Statistical analysis indicated that AKBA 's activity both in the prevention of small intestinal and colonic polyps was more potently than aspirin . Histopathologic examination revealed that AKBA 's effect , that is the reduction of polyp size and degree of dysplasia , was more prominent in larger sized polyps , especially those originating in colon . These effects of AKBA were associated with its role in the induction of apoptosis in carcinomas . The assays of western blotting and immunohistochemistry staining indicated that the efficacy of AKBA might arise from its activity in the modulation of the Wnt/β-catenin pathway and NF-κB/ P35354 pathway in adenomatous polyps . Conclusion , AKBA by oral application prevented intestinal tumorigenesis more potential than aspirin . DB00945 and NSAID sensitivity . DB00945 and the older nonsteroidal anti-inflammatory drugs ( NSAIDs ) that block cyclo-oxygenase-1 ( P23219 ) induce asthma attacks in patients with aspirin-exacerbated respiratory disease and urticaria in patients with chronic idiopathic urticaria . Weak inhibitors of P23219 , such as acetaminophen and salsalate , crossreact also but only with high doses of the drugs . Partial inhibitors of both P23219 and P35354 , such as nimesulide and meloxicam , also cross-react but only at high drug doses . P35354 inhibitors do not cross-react ; however , all NSAIDs , including the selective P35354 inhibitors , can sensitize patients and induce urticaria or anaphylaxis on next exposure to the drug . 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 . The aspirin metabolite , salicylate , inhibits 7,12-dimethylbenz[a]anthracene-DNA adduct formation in breast cancer cells . There is evidence that aspirin and other non-steroidal anti-inflammatory drugs may be protective agents against cancer in the gastrointestinal tract . These effects are particularly well documented for the colon and rectum . Some epidemiological and experimental studies have suggested that aspirin could also be a chemopreventive agent against breast cancer . We investigated the effects of the aspirin metabolite , salicylate ( SA ) , on 7,12-dimethylbenz[a]anthracene ( DMBA ) -DNA adduct formation as well as on the expression of the enzymes involved in the carcinogen bioactivation pathway , in particular cytochrome P450 1A ( CYP1A ) and cyclooxygenases ( P23219 and P35354 ) . The effects of the test drug were examined in both the human mammary carcinoma cell line , MCF-7 , and mammary cells derived from DMBA-induced rat mammary tumours ( RMTCs ) . In this study , we also reported the effects of SA on cell growth and viability in breast cancer cells ( BCCs ) . The results demonstrated that DMBA-DNA adduct formation in both cancer cell lines was inhibited by SA at concentrations of > or = 2.5 mM . CYP1A was undetectable in RMTCs while CYP1A induction by beta-naphthoflavone in MCF-7 cells was significantly inhibited by SA in a concentration-dependent manner . DB00945 did not affect P23219 expression in either of the BCCs . P35354 was not detected in MCF-7 cells , but its expression in RMTCs was inhibited by SA treatment , which also significantly reduced BCC growth , but failed to cause cell death by necrosis or apoptosis . These data suggest that inhibition of DMBA-DNA adduct formation may contribute to aspirin breast cancer chemopreventive action and indicate that this drug can act in the first stage of carcinogenesis . 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 . Peroxisome proliferator-activated receptor-gamma is a target of nonsteroidal anti-inflammatory drugs mediating cyclooxygenase-independent inhibition of lung cancer cell growth . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) inhibit the growth of different cancer cell types , suggesting a broad role for their cyclooxygenase ( P36551 ) targets and eicosanoid products in tumor cell growth . Sulindac sulfide , a P36551 inhibitor , inhibited the growth of non-small-cell lung cancers ( NSCLC ) both in soft agar and as xenografts in nude mice . Importantly , the concentration of sulindac sulfide required to inhibit NSCLC cell growth greatly exceeded the concentration required to inhibit prostaglandin ( PG ) E(2) synthesis in NSCLC cells , suggesting that NSAID inhibition of cell growth is mediated by additional targets distinct from P36551 . Both sulindac sulfide and ciglitazone , a defined peroxisome proliferator-activated receptor-gamma ( PPARgamma ) agonist , stimulated a promoter construct containing a Q07869 response element linked to luciferase and potently inhibited NSCLC cell growth at similar concentrations , indicating a role for PPARgamma as a target of NSAID action in these cells . Overexpression of PPARgamma in NSCLC cells strongly inhibited the transformed growth properties of the cells , providing a molecular confirmation of the results obtained with the PPARgamma agonists . Increased expression of PPARgamma , as well as ciglitazone and sulindac sulfide induced expression of P12830 , which has been linked to increased differentiation of NSCLC . Despite the fact that SCLC cell lines expressed little or no cytosolic phospholipase A(2) , P23219 , or P35354 , sulindac sulfide and PPARgamma agonists also inhibited the transformed growth of these lung cancer cells . We propose that PPARgamma serves as a target for NSAIDs that accounts for P36551 -independent inhibition of lung cancer cell growth . DB00945 triggers formation of anti-inflammatory mediators : New mechanism for an old drug . Extract : DB00945 is a widely used non-steroidal anti-inflammatory drug ( NSAID ) . It is well documented that aspirin irreversibly inhibits cyclooxygenase ( P36551 ) by acetylation of an amino acid serine residue , and thus blocks the subsequent biosynthesis of prostaglandins and thromboxane . P36551 has at least two forms , P23219 and P35354 . P23219 is the main form present in mature platelets in the blood , where it transforms arachidonic acid to the intermediates PG-G/H , which are subsequently converted to thromboxane A2 . Thromboxane A2 is a vasoconstrictor and potent platelet activator . Thus , inhibition of thromboxane A2 formation explains aspirin 's anti-thrombotic properties . In the early 1990s , a second form of P36551 was identified , namely P35354 . P35354 was initially conceptualized as an " inducible " P36551 that is elevated in its quantity by a wide range of agents that stimulate inflammation or cell division and seems to be responsible for local formation during inflammation and cancer . 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 . 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 . 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 . 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 . The Effect of Xuefuzhuyu Oral Liquid on DB00945 Resistance and Its Association with rs5911 , rs5787 , and rs3842788 Gene Polymorphisms . DB00945 should be continued indefinitely in patients after interventional therapy , but 10 % to 40 % of patients experience recurrent vascular events despite adequate aspirin therapy , a condition known as aspirin resistance ( AR ) . Xuefuzhuyu oral liquid , derived from the classic recipe Xuefuzhuyu decoction , has been well documented to inhibit platelet aggregation and to improve hemorheology . The aims of this study were to investigate the effects of Xuefuzhuyu oral liquid on AR in patients with chronic stable angina after percutaneous coronary intervention ( P05154 ) and the possible genetic markers related to the drug response . 43 patients diagnosed as having aspirin resistance or semi-resistance were randomly divided into control and treatment groups after screening 207 stable Q8NE62 patients . Platelet aggregation rate was determined using turbidimetry . Three single nucleotide polymorphisms in P23219 ( rs5787 , rs3842788 ) and GP IIb ( rs5911 ) were genotyped in whole blood samples using ABI Q96FA3 7900 HT Fast Real-Time instrument and ABI Q96FA3 3730 DNA Sequencer . The results showed that Xuefuzhuyu oral liquid could effectively improve blood stasis syndrome and AR by inhibiting ADP-induced platelet aggregation and that patients with the rs5911 genetic variant exhibited better drug response upon treatment with Xuefuzhuyu oral liquid , which suggests Xuefuzhuyu oral liquid as a new possible drug for the prevention of AR . Angioedema associated with aspirin and rofecoxib . OBJECTIVE : To report the probable association of angioedema with aspirin therapy and the selective cyclooxygenase-2 ( P35354 ) inhibitor rofecoxib . CASE SUMMARY : A 44-year-old white woman , previously tolerant to aspirin and other nonsteroidal antiinflammatory drugs ( NSAIDs ) , developed angioedema of the lips after ingesting two 325-mg aspirin tablets during one day . The reaction occurred 3 hours after taking the second aspirin and resolved within 3 hours . Two weeks later , the patient took a 25-mg rofecoxib tablet for a sore throat , and she developed angioedema 5(1/2) hours later . Although the woman took 50 mg of diphenhydramine , the swelling did not subside . She repeated the diphenhydramine dose in the evening and , by noon the next day , 26(1/2) hours after the angioedema began , it was resolved . The patient 's internist prescribed an epinephrine auto-injector and advised her to consult an allergist . With skin testing and oral rechallenge with aspirin , but not rofecoxib , the allergist determined the cause of the reactions to be aspirin-induced angioedema and selective P35354 inhibitor intolerance . The Naranjo probability scale indicated that aspirin was a highly probable cause and rofecoxib was a probable cause of this patient 's angioedema . DISCUSSION : DB00945 -induced angioedema and NSAID intolerance have been well documented . There are reports of both tolerance and intolerance to selective P35354 inhibitors in patients with documented allergy-like reactions to aspirin and NSAIDs . CONCLUSIONS : Patients with aspirin and NSAID intolerance may develop intolerance to P35354 inhibitors , especially with repeated exposure . DB00945 and stroke prevention . According to meta-analyses aspirin provides a relative reduction in the rate of major vascular events of 19 % in patients with arterial disease in general , whereas for patients with ischaemic cerebrovascular disease this reduction is only 13 % . The discrepancy may well result from pathophysiological differences and not from a play of chance . There is no proven difference in efficacy according to dose . The evidence for this equivalence is most compelling in the range between 75 and 1300 mg daily , but still fairly convincing for doses between 30 and 50 mg . In contrast , side effects are clearly more frequent as the dose is higher . Other antiplatelet agents ( sulfinpyrazone , ticlopidine , clopidogrel , dipyridamole , orally administered IIb/IIIa inhibitors ) have no clear advantages over aspirin and in some cases definite disadvantages ; the combination of aspirin and dipyridamole may be more efficacious than aspirin alone , but the evidence hinges on a single trial . If recurrent TIAs occur under treatment with aspirin , the rational response is not to change to a different antiplatelet agent , but to review the diagnosis and consider causes other than artery-to-artery embolism . Platelet aggregation can probably still occur despite complete acetylation of platelets , via pathways other than P23219 inhibition , but in vitro aggregation tests are an unreliable measure . Effect of NSAIDS and P35354 inhibitors on the incidence and severity of asbestos-induced malignant mesothelioma : evidence from an animal model and a human cohort . OBJECTIVES : Non-steroidal anti-inflammatory drugs ( NSAIDs ) and P35354 inhibitors have been associated with lower incidence rates of some cancers . Because asbestos can cause chronic inflammation at the pleural and peritoneal surfaces we hypothesised that NSAID and P35354 inhibitors would inhibit the development of asbestos-induced mesothelioma . MATERIALS AND METHODS : A murine model of asbestos-induced mesothelioma was used to test this hypothesis by providing the NSAID , aspirin , daily in the feed at 50mg/kg or 250 mg/kg . In a parallel study , the relationship between the use of NSAID and P35354 inhibitors and mesothelioma was investigated in a human cohort of 1738 asbestos exposed people living or working in Wittenoom , Western Australia ( a crocidolite mine site ) . RESULTS : DB00945 did not alter the rate of disease development or increase the length of time that mice survived . DB00945 had a small but significant effect on disease latency ( the time between asbestos exposure and first evidence of disease ; p < 0.05 ) but disease progression was not affected by the continued presence of the drug . In the Wittenoom cohort , individuals who reported use of NSAIDs , P35354 inhibitors or both did not have a lower incidence of mesothelioma ( HR=0.85 ; 95 % CI=0.53-1.37 , p=0.50 ) , ( HR=0.69 ; 95 % CI=0.21-2.30 , p=0.55 ) and ( HR=0.43 ; 95 % CI=0.16-1.13 , p=0.087 ) respectively . CONCLUSION : We conclude that NSAIDs and P35354 inhibitors do not moderate mesothelioma development or progression in a human cohort exposed to asbestos and this result is confirmed in an autochthonous mouse model . 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 . Exposure to low doses of formaldehyde during pregnancy suppresses the development of allergic lung inflammation in offspring . DB03843 ( FA ) is an environmental and occupational pollutant , and its toxic effects on the immune system have been shown . Nevertheless , no data are available regarding the programming mechanisms after FA exposure and its repercussions for the immune systems of offspring . In this study , our objective was to investigate the effects of low-dose exposure of FA on pregnant rats and its repercussion for the development of allergic lung inflammation in offspring . Pregnant Wistar rats were assigned in 3 groups : P ( rats exposed to FA ( 0.75 ppm , 1 h/day , 5 days/week , for 21 days ) ) , C ( rats exposed to vehicle of FA ( distillated water ) ) and B ( rats non-manipulated ) . After 30 days of age , the offspring was sensitised with ovalbumin ( OVA ) -alum and challenged with aerosolized OVA ( 1 % , 15 min , 3 days ) . After 24 h the OVA challenge the parameters were evaluated . Our data showed that low-dose exposure to FA during pregnancy induced low birth weight and suppressed the development of allergic lung inflammation and tracheal hyperresponsiveness in offspring by mechanisms mediated by reduced anaphylactic antibodies synthesis , P05231 and P01375 secretion . Elevated levels of P22301 were found . Any systemic alteration was detected in the exposed pregnant rats , although oxidative stress in the uterine environment was evident at the moment of the delivery based on elevated P23219 expression and reduced P29474 and SOD-2 in the uterus . Therefore , we show the putative programming mechanisms induced by FA on the immune system for the first time and the mechanisms involved may be related to oxidative stress in the foetal microenvironment . 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 . 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 . Prostaglandin-endoperoxide synthase genes P23219 and P35354 - novel modifiers of disease severity in cystic fibrosis patients . Cystic fibrosis ( CF ) is one of the most common autosomal recessive diseases among Caucasians caused by a mutation in the P13569 gene . However , the clinical outcome of CF pulmonary disease varies remarkably even in patients with the same P13569 genotype . This has led to a search for genetic modifiers located outside the P13569 gene . The aim of this study was to evaluate the effect of functional variants in prostaglandin-endoperoxide synthase genes ( P23219 and P35354 ) on the severity of lung disease in CF patients . To the best of our knowledge , it is the first time when analysis of P23219 and P35354 as potential CF modifiers is provided . The study included 94 CF patients homozygous for F508del mutation of P13569 . To compare their clinical condition , several parameters were recorded , e.g. a unique clinical score : disease severity status ( DSS ) . To analyse the effect of non- P13569 genetic polymorphisms on the clinical course of CF patients , the whole coding region of P23219 and selected P35354 polymorphisms were analysed . Statistical analysis of genotype-phenotype associations revealed a relationship between the heterozygosity status of identified polymorphisms and better lung function . These results mainly concern P35354 polymorphisms : -765G > C and 8473T > C . The P23219 and P35354 polymorphisms reducing P36551 protein levels had a positive effect on all analysed clinical parameters . This suggests an important role of these genes as protective modifiers of pulmonary disease in CF patients , due to inhibition of arachidonic acid conversion into prostaglandins , which probably reduces the inflammatory process . O76074 inhibitors enhance celecoxib killing in multiple tumor types . The present studies determined whether clinically relevant phosphodiesterase 5 ( O76074 ) inhibitors interacted with a clinically relevant NSAID , celecoxib , to kill tumor cells . Celecoxib and O76074 inhibitors interacted in a greater than additive fashion to kill multiple tumor cell types . Celecoxib and sildenafil killed ex vivo primary human glioma cells as well as their associated activated microglia . Knock down of O76074 recapitulated the effects of O76074 inhibitor treatment ; the nitric oxide synthase inhibitor L-NAME suppressed drug combination toxicity . The effects of celecoxib were P35354 independent . Over-expression of O15519 -s or knock down of CD95/ Q13158 significantly reduced killing by the drug combination . CD95 activation was dependent on nitric oxide and ceramide signaling . CD95 signaling activated the JNK pathway and inhibition of JNK suppressed cell killing . The drug combination inactivated P42345 and increased the levels of autophagy and knock down of Beclin1 or Q9H1Y0 strongly suppressed killing by the drug combination . The drug combination caused an ER stress response ; knock down of IRE1α/ P17861 enhanced killing whereas knock down of eIF2α/ P18848 / P35638 suppressed killing . DB00203 and celecoxib treatment suppressed the growth of mammary tumors in vivo . Collectively our data demonstrate that clinically achievable concentrations of celecoxib and sildenafil have the potential to be a new therapeutic approach for cancer . DB00945 and non-small cell lung cancer resections : effect on long-term survival . OBJECTIVE : Survival after resections for non-small cell lung cancer remains poor . Recurrent lung cancer remains common . Due to the common risk factor of smoking , cardiovascular deaths occur in the absence of recurrent lung cancer in up to 15 % of patients . DB00945 has been proven to reduce cardiovascular mortality as a secondary prophylactic agent , but not as a primary agent . DB00945 being a P35354 inhibitor has been shown to reduce the chance of metastasis in adenocarcinoma but not squamous carcinoma . We sought to investigate the effect of long-term aspirin therapy on survival post potentially curative surgery . METHODS : We analysed a prospective thoracic surgical database , from time period 2003 to date . Patients who were on aspirin pre-operatively , N=412 were compared to non users , N=1353 . Patient long-term outcome was assessed utilising the national strategic tracking service that operates in the United Kingdom . Cox proportional hazards analysis was used to determine significant factors affecting survival . RESULTS : 100 % survival follow up was achieved . Regular users of aspirin had > 5 % increased survival , which was significant , p=0.05 , despite having a higher cardiovascular risk profile . Mode of death data was not available . CONCLUSIONS : Adjuvant aspirin post resection for potentially curative non-small cell lung cancer significantly increases survival . The mechanism of increased survival needs further investigation and is the basis for the trial : Adjuvant DB00945 for Non-Small cell Lung Cancer -- The Big A Trial. www.TheBigATrial.co.uk . 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 . 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 . 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 .

MH_train_1116

MH_train_1116

MH_train_1116

interacts_with DB09036?

multiple_choice

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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 ) . 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 . DB00163 prevents ethanol-induced inflammatory , hormonal , and cytotoxic changes in reproductive tissues . DB00898 causes decreased function of the hypothalamic-pituitary-gonadal ( HPG ) axis . DB00898 resulted in inflammatory changes in HPG manifested by increased concentrations of pro-inflammatory cytokines . Since , such cytokines have deleterious effects on functions of HPG , it seemed possible that ethanol 's suppressive action could be due , at least in part , to this inflammation . Since oxidative stress can cause inflammation , we have used the antioxidant vitamin E to test , whether reducing inflammation might protect reproductive functions from ethanol . Rats were fed an ethanol diet or pair fed identically without ethanol for a 3-week period . For the last 10 days , animals were given 30 IU/kg or 90 IU/kg or vehicle . DB00898 significantly increased hypothalamic , pituitary and testicular P01375 and P05231 , all changes prevented by the higher dose of vitamin E. Also , ethanol induced changes in P01148 , LH , testosterone , and testicular germ cell apoptosis were similarly prevented by vitamin E . These data strikingly show that vitamin E protects the HPG from deleterious effects of ethanol and suggests that the mechanism of this protection might be both anti-inflammatory and antioxidant . 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 . 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 . 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 . Dual P00533 and P42345 targeting in squamous cell carcinoma models , and development of early markers of efficacy . The epidermal growth factor receptor ( P00533 ) is a validated target in squamous cell carcinoma ( SCC ) of the head and neck . Most patients , however , do not respond or develop resistance to this agent . P42345 ( P42345 ) is involved in the pathogenesis of SCC of the head and neck ( SCCHN ) . This study aimed to determine if targeting P42345 in combination with P00533 is effective in SCC , and to develop early pharmacodynamic markers of efficacy . Two SCC cell lines , one resistant ( HEP2 ) and one of intermediate susceptibility ( Detroit 562 ) to P00533 inhibitors , were xenografted in vivo and treated with an P42345 inhibitor ( temsirolimus ) , an P00533 inhibitor ( erlotinib ) or a combination of both . DB06287 exerted superior growth arrest in both cell lines than erlotinib . The combined treatment resulted in synergistic antitumor effects in the Detroit 562 cell line . Immunohistochemical assessment of pharmacodynamic effects in fine-needle aspiration ( FNA ) biopsies early after treatment using phospho MAPK , Phospho-P70 and Ki67 as end points demonstrated pathway abrogation in the Detroit 562 tumours treated with the combination , the only group where regressions were seen . In conclusion , an P42345 inhibitor showed antitumor activity in P00533 -resistant SCC cell lines . Marked antitumor effects were associated with dual pathway inhibition , which were detected by early FNA biopsies . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Inhibition of the signal transducer and activator of transcription-3 ( P40763 ) signaling pathway by 4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylic acid esters . The JAK- P40763 pathway regulates genes that are important in cell proliferation and thus is a promising target for cancer therapy . A high-throughput screening ( HTS ) campaign using an Apo-ONE Homogenous Caspase 3/7 assay in U266 cells identified 4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylic acid ethyl ester 4 as a potential P40763 pathway inhibitor . Optimization of this HTS hit led to the identification of the 7-cyano analogue 8 , which inhibited P40763 -Y705 phosphorylation with an EC 50 of 170 nM . Compound 8 also inhibited cytokine induced JAK activation but did not inhibit P11274 - P00519 activated P42229 phosphorylation in K562 cells .

MH_train_1117

MH_train_1117

MH_train_1117

interacts_with DB08895?

multiple_choice

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 . 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 . Enhanced histone deacetylase enzyme activity in primary myelofibrosis . We measured histone deacetylase ( HDAC ) activity in 17 patients with primary myelofibrosis ( PMF ) ; 19 with other myeloproliferative neoplasm ( Q9BQR3 ) and 16 normal volunteers . Significantly elevated HDAC levels were shown in patients with PMF compared with other Q9BQR3 patients and normal volunteers ( p < 0.05 ) . Sixteen patients with PMF were also studied for correlation between O60674 mutation status and HDAC levels ; no significant correlation was found . We further correlated HDAC levels with clinical features in PMF : there was no correlation with WBC , platelet counts , Hb levels or degree of bone marrow fibrosis , but HDAC levels were correlated to the degree of splenomegaly . This suggests that HDAC may be recruited as essential thrombocythemia or polycythemia vera progresses into myelofibrosis or PMF progresses into more advanced stage . We then used the qRT-PCR cycle threshold ( CT ) method to study which HDACs were elevated in PMF . The results showed that , in general , Class 1 HDACs were elevated ( Q13547 ,2,8 ) except O15379 , Class II HDACs were depressed ( P56524 ,5 ) except Q9UBN7 and 10 , and Class III HDACs were generally elevated . The current study may form the basis for using HDAC inhibitor in the treatment of patients with PMF and may implicate a possible role of HDAC in the association of pathogenesis of PMF . 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 . 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 . 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 . 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 . Signaling pathways implicated in oncostatin M-induced aggrecanase-1 and matrix metalloproteinase-13 expression in human articular chondrocytes . Molecular mechanisms of oncostatin M ( P13725 ) -stimulated cartilage extracellular matrix catabolism and signaling pathways were investigated in human arthritic chondrocytes . P13725 , alone or with Interleukin-1 ( IL-1beta ) , increased glycosaminoglycan release and induced O75173 and P45452 protein expression in human cartilage explants . P13725 dose- and time-dependently increased O75173 mRNA and P45452 protein expression in human femoral head chondrocytes . Extracellular signal-regulated kinases ( P27361 /2 ) -MAPK pathway inhibitor , U0126 , down-regulated O75173 and P45452 induction by P13725 . O60674 ( O60674 ) inhibitor , AG490 , suppressed P13725 -induced O75173 mRNA expression but did not affect P45452 levels while P52333 pharmacological inhibitor and siRNA transfection suppressed both . Parthenolide , a signal transducer and activator of transcription ( P42224 and P40763 ) phosphorylation inhibitor , reduced P13725 -induced O75173 and P45452 gene expression and prevented P42224 /3 DNA binding activity . Additionally , P13725 -enhanced O75173 mRNA and P45452 expression was down-regulated by phosphatidylinositol 3-kinase ( PI3K ) and Akt/ P31749 inhibitors , LY294002 and NL-71-101 . Furthermore , P52333 inhibition time-dependently down-regulated Akt but not P27361 /2 phosphorylation suggesting that Akt is a downstream target of P52333 . These results suggest that P13725 -stimulated O75173 and P45452 expression is mediated by P27361 /2 , P52333 / P42224 /3 and PI3K/Akt and by cross talk between these pathways . The inhibitors of these cascades could block P13725 -evoked degeneration of cartilage by O75173 and P45452 . Characterization of the insulin-signaling pathway in lacrimal and salivary glands of rats . PURPOSE : P01308 has been acknowledged as a mediator of several physiological events in lacrimal and salivary glands . We investigated the presence of insulin receptors and of insulin-induced autophosphorylation of the insulin receptor and activation of elements involved in the early steps of insulin signaling in lacrimal and salivary glands of rats . METHODS : Lacrimal and salivary glands of Wistar rats were removed and processed for immunohistochemistry using anti-insulin receptor and anti- DB01277 receptor antibodies . The activation of insulin receptors following insulin treatment , and the involvement of insulin receptor substrates-1 and -2 , Shc , O60674 and P35610 -1 , were analyzed by immunoprecipitation , followed by SDS-PAGE and immunoblotting of rat lacrimal and salivary glands after exposure to insulin . RESULTS : P01308 and DB01277 receptors were present in rat lacrimal and salivary glands and were located predominantly in the cytoplasm and plasma membrane . Functional studies demonstrated that insulin induced a dose-dependent phosphorylation of the insulin receptor , IGF-1R , insulin receptor substrates-1 and -2 , Shc , and P35610 -1 . In rats with streptozotocin-induced diabetes mellitus there was a significant reduction in insulin-induced insulin receptor and P35610 -1 phosphorylation in the lacrimal gland but not in the salivary gland ; there was no influence on Shc phosphorylation in either tissue . CONCLUSIONS : The present results indicate that insulin and DB01277 receptors are expressed in lacrimal and salivary glands , and that insulin can induce the phosphorylation of its receptor and activate elements involved in the early steps of insulin signaling in both tissues . 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 . P04150 interacting protein-1 restores glucocorticoid responsiveness in steroid-resistant airway structural cells . Glucocorticoid ( GC ) insensitivity represents a profound challenge in managing patients with asthma . The mutual inhibition of transcriptional activity between GC receptor ( GR ) and other regulators is one of the mechanisms contributing to GC resistance in asthma . We recently reported that interferon regulatory factor ( Q969Q1 ) -1 is a novel transcription factor that promotes GC insensitivity in human airway smooth muscle ( P17405 ) cells by interfering with GR signaling ( Tliba et al. , Am J Respir Cell Mol Biol 2008;38:463-472 ) . Here , we sought to determine whether the inhibition of GR function by P10914 involves its interaction with the transcriptional co-regulator GR-interacting protein 1 ( Q9Y3R0 ) , a known GR transcriptional co-activator . We here found that siRNA-mediated Q9Y3R0 depletion attenuated P10914 -dependent transcription of the luciferase reporter construct and the mRNA expression of an P10914 -dependent gene , P28907 . In parallel experiments , Q9Y3R0 silencing significantly reduced GR-mediated transactivation activities . Co-immunoprecipitation and Q86UG4 pull-down assays showed that Q9Y3R0 , through its repression domain , physically interacts with P10914 identifying Q9Y3R0 as a bona fide transcriptional co-activator for P10914 . Interestingly , the previously reported inhibition of GR-mediated transactivation activities by either P01375 and P01579 treatment or P10914 overexpression was fully reversed by increasing cellular levels of Q9Y3R0 . Together , these data suggest that the cellular accumulation of P10914 may represent a potential molecular mechanism mediating altered cellular response to GC through the depletion of Q9Y3R0 from the GR transcriptional regulatory complexes . P05231 and soluble P05231 receptor stimulate the production of MMPs and their inhibitors via JAK- P35610 and P29323 -MAPK signalling in human chondrocytes . Elevated concentrations of P05231 ( interleukin-6 ) and sIL-6r ( soluble P05231 receptor ) in the synovial fluid and serum of patients with arthritis have been implicated in joint cartilage destruction . This study examined the effects of P05231 and sIL-6r on the expression of MMPs ( matrix metalloproteinases ) , TIMPs ( tissue inhibitor of metalloproteinases ) , the plasminogen activation system including tPA ( tissue-type PA ) , uPA ( urokinase-type PA ) and P05121 ( PA inhibitor type 1 ) using chondrocytes derived from normal human femur cartilage . The cells were cultured with or without 50 ng/ml P05231 and/or 30 ng/ml sIL-6r in the presence or absence of the P52333 ( P52333 ) inhibitor WHI-P131 or the MEK [ MAPK ( mitogen-activated protein kinase ) / P29323 ( extracellular signal protein kinase ) kinase ] inhibitor PD98059 for up to 28 days . The expression of MMPs , TIMPs , uPA , tPA and P05121 was investigated at the mRNA and protein levels . MMP protein expression and pSTAT3 ( phosphorylation of signal transducer and activator of transcription 3 ) and pERK ( phosphorylation of P29323 ) were also measured . Treatment with both P05231 and sIL-6r markedly increased the expression of P03956 , P45452 , P01033 and P05121 , while significantly decreasing the expression of tPA and uPA and stimulating pSTAT3 and pERK . Adding WHI-P131 or PD98059 decreased P05231 and sIL-6r enhancement of P03956 , -3 and -13 . The results suggest that P05231 and sIL-6r stimulate the production of MMPs and their inhibitor via JAK- P35610 and P29323 -MAPK signalling in chondrocytes . 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 . [ 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 . 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 . 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 . 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 . DB08820 potentiation of multiple P13569 channels with gating mutations . BACKGROUND : The investigational P13569 potentiator ivacaftor ( VX-770 ) increased P13569 channel activity and improved lung function in subjects with CF who have the G551D P13569 gating mutation . The aim of this in vitro study was to determine whether ivacaftor potentiates mutant P13569 with gating defects caused by other P13569 gating mutations . METHODS : The effects of ivacaftor on P13569 channel open probability and chloride transport were tested in electrophysiological studies using Fischer rat thyroid ( P17948 ) cells expressing different P13569 gating mutations . RESULTS : DB08820 potentiated multiple mutant P13569 forms with defects in P13569 channel gating . These included the G551D , G178R , S549N , S549R , G551S , G970R , G1244E , S1251N , S1255P and G1349D P13569 gating mutations . CONCLUSION : These in vitro data suggest that ivacaftor has a similar effect on all P13569 forms with gating defects and support investigation of the potential clinical benefit of ivacaftor in CF patients who have P13569 gating mutations beyond G551D . Differential regulation of cystic fibrosis transmembrane conductance regulator by interferon gamma in mast cells and epithelial cells . P13569 ( P13569 ) is a P13569 in epithelial cells ; recently , we identified it in mast cells . Previous work that we confirmed showed that interferon gamma ( IFNgamma ) down-regulated P13569 expression in epithelial cells ( T84 ) , but by contrast , we found that IFNgamma up-regulated P13569 mRNA and protein expression in rat and human mast cells . IFNgamma up-regulation of P13569 in mast cells was inhibited by p38 and extracellular signal-regulated kinase ( P29323 ) kinase inhibitors but not a Janus tyrosine kinase (JAK)2 inhibitor , whereas in T84 cells IFNgamma-mediated down-regulation of P13569 was O60674 -dependent and P29323 - and p38-independent . Furthermore , IFNgamma down-regulation of P13569 in T84 epithelial cells was P42224 -dependent , but up-regulation of P13569 in mast cells was P42224 -independent . Thus , differential regulatory pathways of P13569 expression in mast cells and epithelial cells exist that depend upon either p38/ P29323 or JAK/ P35610 pathways , respectively . Surprisingly , IFNgamma treatment of mast cells inhibited Cl(-) efflux , in contrast to up-regulation of P13569 /mRNA and protein expression . However , down-regulation of Cl(-) flux correlated with IFNgamma-mediated inhibition of mediator secretion . This and other work suggests that the effect of IFNgamma on P13569 expression in mast cells is important for their function . 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 . 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 . 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 . 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 . Structure-based design of oxygen-linked macrocyclic kinase inhibitors : discovery of SB1518 and SB1578 , potent inhibitors of O60674 ( O60674 ) and Fms-like tyrosine kinase-3 ( P36888 ) . Macrocycles from our Aurora project were screened in a kinase panel and were found to be active on other kinase targets , mainly JAKs , P36888 and CDKs . Subsequently these compounds became leads in our O60674 project . Macrocycles with a basic nitrogen in the linker form a salt bridge with Asp86 in P24941 and Asp698 in P36888 . This residue is conserved in most CDKs resulting in potent pan CDK inhibition . One of the main project objectives was to achieve O60674 potency with 100-fold selectivity against CDKs . Macrocycles with an ether linker have potent O60674 activity with the ether oxygen forming a hydrogen bond to Ser936 . A hydrogen bond to the equivalent residues of P52333 and most CDKs can not be formed resulting in good selectivity for O60674 over P52333 and CDKs . Further optimization of the macrocyclic linker and side chain increased O60674 and P36888 activity as well as improving Q09013 properties . The selective O60674 / P36888 inhibitor 11 ( Pacritinib , SB1518 ) has successfully finished phase 2 clinical trials for myelofibrosis and lymphoma . Another selective O60674 / P36888 inhibitor , 33 ( SB1578 ) , has entered phase 1 clinical development for the non-oncology indication rheumatoid arthritis . JAK- P35610 and JAK-PI3K-mTORC1 pathways regulate telomerase transcriptionally and posttranslationally in ATL cells . Adult T-cell leukemia ( ATL ) is a heterogeneous tumor that is resistant to chemotherapy . Telomerase activity plays a critical role in tumorigenesis and is associated with the prognosis of ATL patients . Interleukin ( IL ) -2 commonly promotes tumor growth in chronic ATL cells . The signaling pathways involved in P60568 -regulated telomerase activation were studied in ATL cells derived from chronic ATL patients . P60568 challenge enhanced tyrosine phosphorylation of Janus-activated kinase (JAK)1-3 and P42229 , and induced P23458 and O60674 to associate with P42229 in P60568 -dependent ATL cells . Chromatin immunoprecipitation assays revealed that P42229 directly bound to the human telomerase reverse transcriptase ( hTERT ) promoter . P42229 short interfering RNA inhibited hTERT transcription in P60568 -stimulated ATL cells . Inhibitors of PI3K , HSP90 , and P42345 reduced P60568 -induced hTERT mRNA , protein expression , and telomerase activity . AKT , HSP90 , P42345 , S6 kinase , and hTERT immunoprecipitate from P60568 -stimulated cells contained telomerase activity , suggesting that hTERT directly interacts with , and is regulated by , these proteins . Binding of the p85 regulatory subunit of PI3K to O60674 was enhanced in an P60568 -dependent manner , indicating that O60674 propagates activation signals from the P60568 receptor and links hTERT activation to both the P42229 and PI3K pathways . Finally , P60568 -induced activation of telomerase and P42229 was observed in primary leukemic cells . These results indicate that P60568 stimulation induces hTERT activation through the JAK/ P35610 pathway and the JAK/PI3K/AKT/HSP90/mTORC1 pathway in P60568 -responsive ATL cells . These signaling proteins represent novel and promising molecular therapeutic targets for P60568 -dependent ATL . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Human and rodent pancreatic beta-cells express P05112 receptors and P05112 protects against beta-cell apoptosis by activation of the PI3K and JAK/ P35610 pathways . Secretion of pro-inflammatory cytokines is associated with loss of pancreatic beta-cell viability and cell death . P05112 ( interleukin-4 ) has been reported to mediate a protective effect against the loss of pancreatic beta-cells , and P05112 receptors have been found in rat pancreatic beta-cells at both the RNA and the protein level . The aim of the present study was to investigate P05112 receptor expression in human islet cells and to examine the signalling pathways by which P05112 exerts its effects using the rat beta-cell lines , BRIN-BD11 and P01308 -1E . By means of immunohistochemistry , it was demonstrated that P05112 receptors are present on human islet cells . Using a flow cytometric method for evaluating cell death , it was confirmed that incubating beta-cells with P05112 attenuated cell death induced by IL-1beta and interferon-gamma by approx . 65 % . This effect was abrogated by the presence of the PI3K ( phosphoinositide 3-kinase ) inhibitor , wortmannin , suggesting that activation of the PI3K pathway is involved . In support of this , Western blotting revealed that incubation of cells with P05112 resulted in increased phosphorylation of Akt ( also called protein kinase B ) , a downstream target of PI3K . Increased tyrosine phosphorylation of P42226 ( signal transducer and activator of transcription 6 ) also occurred in response to P05112 and a selective P52333 ( P52333 ) inhibitor reduced the cytoprotective response . Both effects were prevented by overexpression of the tyrosine phosphatase , PTP-BL ( protein tyrosine phosphatase-BL ) . We conclude that P05112 receptors are functionally competent in pancreatic beta-cells and that they signal via PI3K and JAK/ P35610 pathways . These findings may have implications for future therapeutic strategies for the management of diabetes . 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 . 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 . Identification of receptor genes in renal cell carcinoma associated with angiogenesis by differential hybridization technique . To screen the receptor genes in renal cell carcinoma ( RCC ) associated with angiogenesis , we performed differential hybridization of the cDNA library of membrane-type protein tyrosine kinases ( mPTKs ) . Three thousand plaques of a mPTKs-enriched cDNA library were screened with mPTKs mixture probes produced from hypervascular RCC tissues and RCC cell lines . Six different cDNA fragments of the PTK genes were isolated , and the sequence analysis showed that these represented cDNAs for P35590 , P35968 , P07333 , P22455 , P23458 and P08631 . Of these genes , the expression of P35590 , P35968 , and P22455 was studied in RCC tissue and cell lines by Northern blot analysis . We also investigated the expression of vascular endothelial growth factor ( P15692 ) , placenta growth factor ( P49763 ) and their receptor P17948 . In all the hypervascular RCC tissues , the amounts of mRNAs for P35968 and P17948 were increased compared to adjacent normal tissues . The P35590 and P22455 genes were also overexpressed in most of the hypervascular RCC tissues , while no mRNA of P35968 , P17948 , or P35590 could be detected in any of the four human RCC cell lines . The amounts of the P15692 and P49763 mRNAs were increased in hypervascular RCC tissues , while P15692 mRNA was detected in the four cell lines but P49763 mRNA was not . P22455 mRNA was expressed in three of the four cell lines . These results suggest that P35968 , P17948 , P49763 and P35590 mRNAs are present in the mesenchymal cells of RCC , while P15692 and P22455 genes are expressed in RCC cells themselves in vivo . Mechanisms of Nattokinase in protection of cerebral ischemia . In vivo , the level of cyclic DB00640 Monophosphate ( DB02527 ) and the pathway of the Janus Kinase1/Signal Transducers and Activators of Transcription1 ( P23458 / P42224 ) were studied . In vitro , the Ca(2+) mobilization in human platelet stimulated by thrombin was observed . In addition , vasomotion of vascular smooth muscle was measured by adding DB00761 or norepinephrine(NE) under the Ca(2+) contained bath solutions . The effect induced by NE in the presence of N-nitro-L-arginine methyl ester ( L-NAME ) or indometacin ( Indo ) was also detected . At last , the levels of tissue plasminogen activator ( t-PA ) and P00747 activator inhibitor-1 ( P05121 ) in cultured supernatans in Human umbilical vein endothelial cells ( Huvecs ) were measured by means of ELISA kit . Results showed that Nattokinase ( NK ) significantly increased the DB02527 level , activated the signal passage of P23458 / P42224 in injured part and inhibited remarkably the rise of platelet intracellular Ca(2+) ( [Ca(2+)]i ) in human platelet . Furthermore , NK relaxed rat thoracic aortic artery in the dose-dependent manner and in the endothelium dependent manner and its effect could be attenuated by L-NAME . Also , the secretion of t-PA and P05121 were reduced stimulated by Adr on Huvecs . These data indicated that the neuroprotective effect of NK was associated with its antiplatelet activity by elevating DB02527 level and attenuating the calcium release from calcium stores ; with its anti-apoptotic effect through the activation of P23458 / P42224 pathway ; with its relaxing vascular smooth muscle by promoting synthesis and release of NO , reducing ROC calcium ion influx and with its protection on endothelial cells through increasing fibrinolytic activity and facilitating spontaneous thrombolysis . 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 . 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 . Transcriptional profiles during the differentiation and maturation of monocyte-derived dendritic cells , analyzed using focused microarrays . Dendritic cells ( DC ) are professional antigen-presenting cells capable of initiating primary immune responses . They have been intensively studied and are used in both basic immunology research and clinical immunotherapy . However , the genetic pathways leading to DC differentiation and maturation remain poorly understood . Using focused microarrays with oligonucletotide probes for 120 genes encoding co-stimulatory molecules , chemokines , chemokine receptors , cytokines , cytokine receptors , TLRs , and several other related molecules , we analyzed the kinetics of gene expression for the overall differentiation process of monocytes into mature DC . In parallel , we compared the transcriptional profiles in DC maturation in the presence of LPS , P01375 or trimeric P29965 . We found similar transcriptional profiles for early immature DC and immature DC , respectively generated by culturing monocytes with GM- P04141 and P05112 for three or six days . We identified sets of common and stimuli-specific genes , the expression of which changed following stimulation with LPS , P01375 or P29965 . A dynamic analysis of the entire DC differentiation and maturation process showed that some important inflammatory and constitutive chemokines are transcribed in both immature and mature DC . The correlative expression kinetics of the gene pairs P14778 / P27930 , P40933 / Q13261 , Q9NNX6 / P13598 and Q9NNX6 / P32942 imply that they all play crucial roles in mediating DC functions . Thus , our analysis with focused microarrays shed light on the transcriptional kinetics of DC differentiation and maturation , and this method may also prove useful for identifying novel marker genes involved in DC functions . 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 . 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 . 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) . Serotonin receptor activation leads to neurite outgrowth and neuronal survival . Serotonin 5-HT1 receptors are implicated in anxiety and depression . These receptors belong to the family A of G-protein-coupled receptors and couple to inhibitory G-proteins . Recent studies show that chronic activation of P08908 receptors leads to proliferation of hippocampal neurons suggesting that neurogenesis contributes to the effects of antidepressants . However , the molecular mechanisms and pathways involved are not understood . We used Neuro 2A cells transfected with P08908 receptors and SK-N-SH cells endogenously expressing the receptor to examine the effect of receptor activation on neuronal survival and neurite outgrowth . We find that receptor activation leads to increased neurite outgrowth that can be blocked by the receptor selective antagonist and by treatment with pertussis toxin or lactacystin implicating inhibitory G-proteins and proteasomal degradation in this process . Interestingly , the small G-protein Rap and the transcription factor P35610 -3 are also involved since reducing the levels of Rap1 ( using small interfering RNA ) or P35610 -3 ( using dominant negative P40763 ) significantly blocks P08908 -receptor-mediated neurite outgrowth . The observed increase in the phosphorylation of Src and P35610 -3 , at sites leading to their activation , further supports a crucial role for these proteins in neurite outgrowth . We also find that prolonged activation of endogenous P08908 receptors leads to increased cell survival even under starving conditions ; this is completely blocked by co-treatment with the antagonist . Taken together , these findings indicate that activation of the P08908 receptor leads to a number of neurotropic events by activating a series of signal transduction molecules leading to long-term changes required for neurogenesis . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . 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 . P13569 mutations impart elevated immune reactivity in a murine model of cystic fibrosis related diabetes . Increased life expectancy in cystic fibrosis ( CF ) is accompanied by an increasing incidence of CF related diabetes ( CFRD ) . Altered immune reactivity occurs in CF , which we hypothesize , is exacerbated by hyperglycemia . P13569 deficient ( P13569 -/- ) mice were rendered hyperglycemic by streptozotocin ( Q11206 ) to test this hypothesis . P13569 -/- , C57BL/6J , and FVB/NJ mice received either Q11206 or lactated ringers ( LR ) ( n=5-10 ) . Four weeks later , splenocytes were harvested , mitogen stimulated , and analyzed for cytokine production ( P60568 , P05112 , and P22301 ) along with stimulation indices ( SI ) . SI of Q11206 -treated P13569 -/- were elevated compared to LR-treated mice , although both were greater than C57BL/6J and FVB/NJ ( p < 0.05 ) . Fasting glucose levels of Q11206 -treated P13569 -/- mice correlated with SI ( p < 0.003 ) . Stimulated P22301 concentrations were elevated in Q11206 -treated P13569 -/- compared to LR-treated animals and controls ( p < 0.05 ) . P60568 levels were greater in P13569 -/- mice compared to controls ( p < 0.05 ) , but unrelated to Q11206 . Reinforcing generalized cytokine up-regulation in P13569 -/- , P05112 levels were greater in P13569 -/- mice compared to C57BL/6J , but FVB/NJ mice demonstrated greatest concentrations following Q11206 . These results suggest that , hyperglycemia may exacerbate the clinical course in CF by impacting immune reactivity . There is clear need to maximize metabolic management in CFRD . P52333 activity is necessary for phosphorylation of cytosolic phospholipase A2 and prostaglandin E2 synthesis by macrophages infected with Francisella tularensis live vaccine strain . Francisella tularensis , the causative agent of tularemia , modulates the host immune response to gain a survival advantage within the host . One mechanism of immune evasion is the ability of F. tularensis to induce the synthesis of the small lipid mediator prostaglandin E2 ( DB00917 ) , which alters the host T cell response making the host more susceptible to Francisella growth . DB00917 is synthesized by a tightly regulated biosynthetic pathway following stimulation . The synthesis of DB00917 begins with the liberation of arachidonic acid ( AA ) from membrane phospholipids by cytosolic phospholipase A2 ( P47712 ) . AA is subsequently converted to the unstable intermediate PGH2 by cyclooxygenase-2 ( P35354 ) , and PGH2 undergoes an isomerization reaction to generate DB00917 . Our objective was to identify F. tularensis-activated host signaling pathways that regulate the activity of the enzymes in the DB00917 -biosynthetic pathway . In this study , we show that P47712 , p38 mitogen-activated protein kinase ( MAPK ) , and P52333 ( P52333 ) signaling are necessary for F. tularensis-induced DB00917 production . Inhibition of P52333 activity reduced the phosphorylation of P47712 and P35354 protein levels . In addition , P52333 regulates P47712 phosphorylation independent of transcription . Moreover , p38 MAPK activity is required for F. tularensis-induced P35354 protein synthesis , but not for the phosphorylation of P47712 . This research highlights a unique signaling axis in which P52333 and p38 MAPK regulate the activity of multiple enzymes of the DB00917 -biosynthetic pathway in macrophages infected with F. tularensis . An P12821 inhibitor reduces Th2 cytokines and TGF-beta1 and TGF-beta2 isoforms in murine lupus nephritis . BACKGROUND : P12821 ( P12821 ) inhibitors , such as captopril , are used to control hypertension . In patients and animals with primary nephropathies , these agents improve renal function more than that would be expected from their control of hypertension . Here , we examine the effects of treatment with captopril on lupus nephritis and discuss the potential mechanism(s) by which this agent exerts its renoprotective effects . METHODS : Lupus-prone , NZB/NZW F1 and MRL-lpr/lpr , mice were treated with captopril or with a control antihypertensive agent , verapamil . Mice were monitored for nephritis , and their sera and tissues analyzed for cytokine and transforming growth factor-beta ( TGF-beta ) expression . RESULTS : DB01197 treatment delayed the onset of proteinuria when administered to prenephritic mice , whereas verapamil did not . DB01197 treatment also retarded disease progression when given to lupus mice that had early disease , and even reversed severe proteinuria in at least some older animals with advanced disease . It reduced chronic renal lesions , but had no effect on autoantibody production . The improvement in renal disease correlated with reduced TGF-beta expression , particularly of the TGF-beta1 and TGF-beta2 isoforms , in the kidneys . Interestingly , in vivo or in vitro exposure to captopril reduced splenic levels of type 2 cytokines , interleukin ( IL ) -4 and P22301 , suggesting a possible role of the immune system in captopril-mediated disease modulation . CONCLUSION : Since type 2 cytokines are known to promote lupus glomerulosclerosis , decreased P05112 and P22301 production in captopril-treated mice may be related to this agent 's renoprotective effects . We argue here that P12821 inhibitors not only act as selective TGF-beta inhibitors , but also as selective immunomodulators , to improve lupus nephritis . 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 ' . OSU-03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier : Implications for Anti-Cancer Therapies . We examined the interaction between OSU-03012 ( also called AR-12 ) with phosphodiesterase 5 ( O76074 ) inhibitors to determine the role of the chaperone glucose-regulated protein ( P11021 ) / P11021 / P11021 in the cellular response . DB00203 ( Viagra ) interacted in a greater than additive fashion with OSU-03012 to kill stem-like GBM cells . Treatment of cells with OSU-03012/sildenafil : abolished the expression of multiple oncogenic growth factor receptors and plasma membrane drug efflux pumps and caused a rapid degradation of P11021 and other HSP70 and HSP90 family chaperone proteins . Decreased expression of plasma membrane receptors and drug efflux pumps was dependent upon enhanced Q9NZJ5 -eIF2α- P18848 - P35638 signaling and was blocked by P11021 over-expression . In vivo OSU-03012/sildenafil was more efficacious than treatment with celecoxib and sildenafil at killing tumor cells without damaging normal tissues and in parallel reduced expression of P08183 and Q9UNQ0 in the normal brain . The combination of OSU-03012/sildenafil synergized with low concentrations of sorafenib to kill tumor cells , and with lapatinib to kill P00533 over-expressing tumor cells . In multiplex assays on plasma and human tumor tissue from an OSU-03012/sildenafil treated mouse , we noted a profound reduction in uPA signaling and identified FGF and P23458 /2 as response biomarkers for potentially suppressing the killing response . Inhibition of FGFR signaling and to a lesser extent P23458 /2 signaling profoundly enhanced OSU-03012/sildenafil lethality . 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 . Drosophila Answers to Q13148 Proteinopathies . Initially implicated in the pathogenesis of P13569 and HIV-1 transcription , nuclear factor Q13148 was subsequently found to be involved in the origin and development of several neurodegenerative diseases . In 2006 , in fact , it was reported for the first time the cytoplasmic accumulation of Q13148 in ubiquitin-positive inclusions of P35858 and FTLD patients , suggesting the presence of a shared underlying mechanism for these diseases . Today , different animal models of Q13148 proteinopathies are available in rodents , nematodes , fishes , and flies . Although these models recapitulate several of the pathological features found in patients , the mechanisms underpinning the progressive neuronal loss observed in Q13148 proteinopathies remain to be characterized . Compared to other models , Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies . At present , the development of Q13148 -related Drosophila models has further strengthened the hypothesis that both Q13148 " loss-of-function " and " gain-of-function " mechanisms can contribute to disease . The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies , along with the information they have generated , towards a better understanding of the mechanisms underlying Q13148 proteinopathies . 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 . 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 . 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 . P60568 regulates expression of C- O75444 in human P01730 T cells . Blockade of IL-2R with humanized anti-CD25 Abs , such as daclizumab , inhibits Th2 responses in human T cells . Recent murine studies have shown that P60568 also plays a significant role in regulating Th2 cell differentiation by activated P42229 . To explore the role of activated P42229 in the Th2 differentiation of primary human T cells , we studied the mechanisms underlying P60568 regulation of C- O75444 expression . Chromatin immunoprecipitation studies revealed that P60568 induced P42229 binding to specific sites in the C- O75444 promoter . These sites corresponded to regions enriched for markers of chromatin architectural features in both resting P01730 and differentiated Th2 cells . Unlike P05231 , P60568 induced C- O75444 expression in P01730 T cells with or without prior TCR stimulation . TCR-induced C- O75444 expression was significantly inhibited by treatment with daclizumab or a P52333 inhibitor , R333 . Furthermore , P60568 and P05231 synergistically induced C- O75444 expression in TCR-activated T cells , suggesting functional cooperation between these cytokines . Finally , both TCR-induced early P05112 mRNA expression and P05112 cytokine expression in differentiated Th2 cells were significantly inhibited by IL-2R blockade . Thus , our findings demonstrate the importance of P60568 in Th2 differentiation in human T cells and support the notion that IL-2R-directed therapies may have utility in the treatment of allergic disorders . 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 . 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 . 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 . DNA replication stalling attenuates tyrosine kinase signaling to suppress S phase progression . Here we report that T cell protein tyrosine phosphatase ( P17706 ) -dependent and -independent pathways attenuate the JAK and Src protein tyrosine kinases ( PTKs ) and P40763 phosphorylation to suppress cyclin D1 expression and S phase progression in response to DNA replication stress . Cells that lack P17706 fail to suppress P23458 , Src , and P40763 , allowing for sustained cyclin D1 levels and progression through S phase despite continued replication stress . Cells that bypass the checkpoint undergo aberrant mitoses with lagging chromosomes that stain for the DNA damage marker gamma P16104 . Therefore , inactivating JAK , Src , and P40763 signaling pathways in response to DNA replication stress may be essential for the suppression of S phase progression and the maintenance of genomic stability .

MH_train_1118

MH_train_1118

MH_train_1118

interacts_with DB00208?

multiple_choice

Cathelicidin LL-37 induces time-resolved release of LTB4 and TXA2 by human macrophages and triggers eicosanoid generation in vivo . In humans , LL-37 and eicosanoids are important mediators of inflammation and immune responses . Here we report that LL-37 promotes leukotriene B4 ( LTB4 ) and thromboxane A2 ( TXA2 ) generation by human monocyte-derived macrophages ( HMDMs ) . LL-37 evokes calcium mobilization apparently via the Q99572 receptor ( P2X7R ) , activation of P27361 /2 and p38 MAPKs , as well as cytosolic phospholipase A2 ( P47712 ) and P09917 in HMDMs , leading to an early ( 1 h ) release of LTB4 . Similarly , TXA2 production at an early time involved the same signaling sequence along an LL-37-P2X7R- P47712 -cyclooxygenase-1 ( P23219 ) axis . However , at later ( 6-8 h ) time points , internalized LL-37 up-regulates P35354 expression , promoting TXA2 production . Furthermore , intraperitoneal injection of mice with murine cathelicidin-related antimicrobial peptide ( mCRAMP ) induces significantly higher levels of LTB4 and TXA2 in mouse ascites rich in macrophages . Conversely , cathelicidin-deficient ( Cnlp(-/-) ) mice produce much less LTB4 and TXB2 in vivo in response to P01375 -α compared with control mice . We conclude that LL-37 elicits a biphasic release of eicosanoids in macrophages with early , Ca(2+)-dependent formation of LTB4 and TXA2 followed by a late peak of TXA2 , generated via induction of P35354 by internalized LL-37 , thus allowing eicosanoid production in a temporally controlled manner . Moreover , our findings provide evidence that LL-37 is an endogenous regulator of eicosanoid-dependent inflammatory responses in vivo . Purinergic receptor-mediated effects of adenosine 5'-triphosphate in urological malignant diseases . DB00171 ( DB00171 ) mediates a variety of biological functions and has been shown to play a physiological role in almost every system in the body . In the genito-urinary system , extracellular DB00171 has been shown to play a functional role in several different capacities , ranging from nociception in the ureter and bladder , to erectile dysfunction via its action on different ' purinergic receptors ' . Discovery of the trophic effects of DB00171 has led to a surge in interest in this signalling system in various malignancies . To date five P2 receptor subtypes have been implicated in the growth inhibition of cancer cells , namely Q93086 , Q99572 , P47900 , P41231 and Q96G91 . Limited data are available on urological malignancies . DB00171 induces its anti-neoplastic effect primarily via purinergic receptor-mediated apoptosis via calcium-independent pathways , and this has been confirmed in vitro and in vivo . Studies have highlighted functional roles for the Q93086 and/or Q96G91 receptors in both hormone refractory prostate cancer and high-grade bladder cancer , although the contributory effect of pro-apoptotic Q99572 receptors remains unclear . Clinical trials have shown intravenous DB00171 successfully attenuates a range of systemic symptoms associated with advanced malignancies . This raises the possibility that selective targeting of specific aberrant pathways may allow for treatment of advanced primary malignancies and their systemic effects . 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 . 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 . Amplification of human platelet activation by surface pannexin-1 channels . BACKGROUND : Q96RD7 ( Panx1 ) forms an anion-selective channel with a permeability up to ~1 kDa and represents a non-lytic , non-vesicular DB00171 release pathway in erythrocytes , leukocytes and neurons . Related connexin gap junction proteins have been reported in platelets ; however , the expression and function of the pannexins remain unknown . OBJECTIVE : To determine the expression and function of pannexins in human plate-lets , using molecular , cellular and functional techniques . METHODS : Panx1 expression in human platelets was det-ermined using qPCR and antibody-based techniques . Contributions of Panx1 to agonist-evoked efflux of cytoplasmic calcein , Ca(2+) influx , DB00171 release and aggregation were assessed in washed platelets under conditions where the P51575 receptor response was preserved ( 0.32 U mL(-1) apyrase ) . Thrombus formation in whole blood was assessed in vitro using a shear chamber assay . Two structurally unrelated and widely used Panx1 inhibitors , probenecid and carbenoxolone , were used throughout this study , at concentrations that do not affect connexin channels . RESULTS : Q96RD7 , but not Q96RD6 or Q96QZ0 , mRNA was detected in human platelets . Furthermore , Panx1 protein is glycosylated and present on the plasma membrane of platelets , and displays weak physical association with P51575 receptors . Panx1 inhibition blocked thrombin-evoked efflux of calcein , and reduced Ca(2+) influx , DB00171 release , platelet aggregation and thrombus formation under arterial shear rates in vitro . The Panx1-dependent contribution was not additive to that of P51575 receptors . CONCLUSIONS : Panx1 is expressed on human platelets and amplifies Ca(2+) influx , DB00171 release and aggregation through the secondary activation of P51575 receptors . We propose that Panx1 represents a novel target for the management of arterial thrombosis . Adaptive phenotype of microglial cells during the normal postnatal development of the somatosensory " Barrel " cortex . Accumulative evidence indicates that microglial cells influence the normal development of central nervous system ( CNS ) synapses . Yet , the functional properties of microglia in relation with synapse development remain unclear . We recently showed that in layer 4 of the whisker-related barrel field of the mouse somatosensory cortex , microglial cells are recruited only after postnatal day (P)5 in the center of the barrels where thalamo-cortical synapses are concentrated and begin their maturation . In the present study , we analyzed the phenotype of microglia during this developmental process . We show that between Q15084 and Q0GE19 microglial cells acquire a more ramified morphology with a smaller soma , they express classical markers of microglia ( Iba1 , CD11b , and P34810 ) but never markers of activation ( Mac-2 and MHCII ) and rarely the proliferation marker Ki67 . Electrophysiological recordings in acute cortical slices showed that at Q15084 a proportion of layer 4 microglia transiently express voltage-dependant potassium currents of the delayed rectifier family , mostly mediated by Kv1.3 subunits , which are usually expressed by activated microglia under pathological conditions . This proportion of cells with rectifying properties doubles between Q15084 and P6 , in concomitance with the beginning of microglia invasion of the barrel centers . Finally , analysis of the responses mediated by purinergic receptors indicated that a higher percentage of rectifying microglia expressed functional Q15077 and Q9H244 receptors , as compared with nonrectifying cells , whereas all cells expressed functional Q99572 receptors . Our results indicate that during normal cortical development distinct microglia properties mature differentially , some of them being exquisitely influenced by the local environment of the maturating neuronal network . 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 . 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 . 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 . Oxidized DB00171 ( oATP ) attenuates proinflammatory signaling via P2 receptor-independent mechanisms . Periodate-oxidized DB00171 ( oATP ) , which covalently modifies nucleotide-binding proteins , can significantly attenuate proinflammatory signaling . Although the Q99572 nucleotide receptor ( P2X7R ) is irreversibly antagonized by oATP , it is unclear whether anti-inflammatory actions of oATP are predominantly mediated via its actions on P2X7R . Here , we describe inhibitory effects of oATP on proinflammatory responses in three human cell types that lack expression of P2X7R : human umbilical vein endothelial cells ( HUVEC ) , HEK293 cells , and 1321N1 astrocytes . oATP decreased by 40-70 % the secretion of interleukin ( IL ) -8 stimulated by tumor necrosis factor-alpha ( P01375 ) in all three cell types , by IL-1beta in HUVEC and 1321N1 cells , and by endotoxin in HUVEC . Attenuation of P01375 -stimulated P10145 secretion by oATP was similar in wild-type P29320 cells or P29320 cells stably expressing recombinant P2X7R . oATP also attenuated cytokine-stimulated expression of nuclear factor-kappaB-luciferase reporter genes expressed in P29320 or 1321N1 cells , but did not affect the rapid downregulation of IkappaB. oATP had no effect on uridine triphosphate-induced activation of native P41231 receptors in P29320 cells , but reduced the potency and efficacy of ADP as an agonist of native P47900 receptors . However , inhibition of P47900 receptors with the specific antagonist MRS2216 did not mimic the effects of oATP on P01375 -stimulated P10145 secretion . Although 1321N1 astrocytes lack expression of any known P2 receptor subtypes , oATP markedly inhibited ecto-ATPase activity in these cells , resulting in a significant accumulation of extracellular DB00171 . In summary , oATP can attenuate proinflammatory signaling by mechanisms independent of the expression or activation of known P2 receptor subtypes . Role of phosphoinositide 3-kinase beta in platelet aggregation and thromboxane A2 generation mediated by Gi signalling pathways . PI3Ks ( phosphoinositide 3-kinases ) play a critical role in platelet functional responses . PI3Ks are activated upon Q9H244 receptor stimulation and generate pro-aggregatory signals . Q9H244 receptor has been shown to play a key role in the platelet aggregation and thromboxane A2 generation caused by co-stimulation with Gq or Gz , or super-stimulation of Gi pathways . In the present study , we evaluated the role of specific PI3K isoforms alpha , beta , gamma and delta in platelet aggregation , thromboxane A2 generation and P29323 ( extracellular-signal-regulated kinase ) activation . Our results show that loss of the PI3K signal impaired the ability of ADP to induce platelet aggregation , P29323 phosphorylation and thromboxane A2 generation . We also show that Gq plus Gi- or Gi plus Gz-mediated platelet aggregation , P29323 phosphorylation and thromboxane A2 generation in human platelets was inhibited by O43548 -221 , a P42338 -selective inhibitor , but not by PIK75 ( a P42336 inhibitor ) , AS252424 ( a P48736 inhibitor ) or IC87114 ( a O00329 inhibitor ) . O43548 -221 also showed a similar inhibitory effect on the Gi plus Gz-mediated platelet responses in platelets from P47900 -/- mice . Finally , 2MeSADP ( 2-methyl-thio-ADP ) -induced Akt phosphorylation was significantly inhibited in the presence of O43548 -221 , suggesting a critical role for P42338 in Gi-mediated signalling . Taken together , our results demonstrate that P42338 plays an important role in ADP-induced platelet aggregation . Moreover , P42338 mediates ADP-induced thromboxane A2 generation by regulating P29323 phosphorylation . [ 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 . 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 . 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 . 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 . 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 . 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 . 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 . [ 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 . Direct oral anticoagulants in acute coronary syndrome . Patients with acute coronary syndromes ( ACS ) require a specific antithrombotic therapy in the immediate and the post ACS phase . The current antithrombotic therapy in the acute phase of an ACS combines antiplatelet and anticoagulant drugs in order to reduce ischemic cardiovascular events . In the post ACS phase , dual antiplatelet therapy ( DAPT ; aspirin and a Q9H244 receptor antagonist ) is the current mainstay of antithrombotic treatment and is recommended in the guidelines of the major North American and European clinical cardiology associations ( DB00551 , ACC , and ESC ) . Recently , the addition of rivaroxaban , a low dose oral direct factor Xa inhibitor ( 2.5 mg twice daily ) , to DAPT ( aspirin plus second-generation Q9H244 inhibitor ) showed a significant reduction of cardiovascular and overall mortality in the major phase III clinical trial ATLAS ACS 2 TIMI 51 . This led to the approval of low-dose rivaroxaban in addition to aspirin and clopidogrel by the European Medicines Agency ( P15941 ) in 2013 . Other direct oral anticoagulants ( apixaban , dabigatran etexilate ) have also been assessed in phase II ( dabigatran etexilate ) and phase III ( apixaban ) post ACS clinical trials . In the studied dosing regimens , these drugs failed to show a net clinical benefit in addition to dual antiplatelet therapy . The major clinical phase II and III post ACS studies of direct oral anticoagulants are summarized and discussed in this article along with the concept of long-term anticoagulation for the secondary prevention of ischemic events after ACS and implications for the future of antithrombotic therapy in the current era of third-generation Q9H244 receptor inhibitors ( Prasugrel and DB08816 ) . Autocrine regulation of γ-irradiation-induced DNA damage response via extracellular nucleotides-mediated activation of Q15077 and Q9H244 receptors . A key component of the response to DNA damage caused by ionizing radiation is DNA repair . Release of extracellular nucleotides , such as DB00171 , from cells plays a role in signaling via P2 receptors . We show here that release of DB00171 , followed by activation of P2Y receptors , is involved in the response to γ-irradiation-induced DNA damage . Formation of phosphorylated histone variant P16104 ( γ P16104 ) foci , which are induced in nuclei by DNA damage and contribute to accumulation of DNA-repair factors , was increased at 1-3h after γ-ray irradiation ( 2.0Gy ) of human lung cancer A549 cells . Focus formation was suppressed by pre-treatment with the ecto-nucleotidase apyrase . Pre-treatment with ecto-nucleotidase inhibitor ARL67156 or post-treatment with DB00171 or UTP facilitated induction of γ P16104 , indicating that extracellular nucleotides play a role in induction of γ P16104 foci . Next , we examined the effect of P2 receptor inhibitors on activation of ataxia telangiectasia mutated ( Q13315 ; a protein kinase ) and accumulation of Q12888 ( a DNA repair factor ) , both of which are important for DNA repair , at DNA damage sites . Q15077 receptor antagonist MRS2578 , Q9H244 receptor antagonist clopidogrel , and Q99572 receptor antagonists A438079 and oxATP significantly inhibited these processes . Release of DB00171 was detected within 2.5min after irradiation , but was blocked by A438079 . Activation of Q13315 and accumulation of Q12888 were decreased in Q15077 or Q9H244 receptor-knockdown cells . We conclude that autocrine/paracrine signaling through Q99572 -dependent DB00171 release and activation of Q15077 and Q9H244 receptors serves to amplify the cellular response to DNA damage caused by γ-irradiation . 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 . 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 . 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 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 . 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 . 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 . 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 . [ 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 . Differential effects of P47900 and Q9H244 nucleotide receptors on P27361 / P28482 and phosphatidylinositol 3-kinase signalling and cell proliferation in serum-deprived and nonstarved glioma P13671 cells . We have previously shown that , in glioma P13671 cells , two nucleotide ADP-sensitive receptors coexist : P47900 , coupled to P98160 and responsible for Ca2+ release , and Q9H244 , negatively coupled to adenylate cyclase . In the present study , we examined the effects of the stimulation of these two receptors on P27361 /2 and P19957 -K activation , and cell proliferation in either serum-deprived or nonstarved P13671 cells . In response to ADP and its analogues , in serum-starved cells , both Q8TCB0 P27361 and Q8NFH3 P28482 were activated in a time-dependent manner , as monitored by Western blot analysis using an antiphospho- Q8NFH3 / Q8TCB0 MAPK antibody . The phosphorylation was reduced both by removal of the extracellular Ca2+ and partially or almost completely by MRS2179 or AR-C69931MX , specific antagonists of the P47900 and Q9H244 receptors , respectively . The inhibitory effect of antagonists was additive . These data indicate the involvement of both receptors , P47900 and Q9H244 , in the P27361 /2 activation , but the Q9H244 receptor contribution predominates . P27361 /2 activity was positively correlated with cell proliferation of cultured glioma P13671 cells . In nonstarved cells , ADP markedly decreased the P19957 -K activity . In contrast , in serum-starved cells , ADP evoked an increase in the P19957 -K activity . Blocking of the P47900 receptor by MRS2179 additionally increased this ADP response . These results suggest that the P47900 receptor has an inhibitory and the Q9H244 receptor a stimulatory effect on P19957 -K signalling pathway . RT-PCR analysis revealed different mRNA expression of both receptors in starved and nonstarved cells . In nonstarved cells , the P47900 receptor mRNA predominates , whereas in serum-deprived cells the expression of Q9H244 mRNA becomes more pronounced . British Journal of Pharmacology ( 2004 ) 141 , 497-507. doi:10.1038/sj.bjp.0705639 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 . DB00107 increases invasive properties of endometrial cancer cells through phosphatidylinositol 3-kinase/AKT-dependent up-regulation of cyclooxygenase-1 , -2 , and P98170 . Traditionally , oxytocin ( OT ) is well known to play a crucial role in the regulation of cyclic changes in the uterus , implantation of the embryo , and parturition . Recently , an additional role for OT has been identified in several types of cancer cells in which OT acts as a growth regulator . In endometrial cancer cells , OT is known to efficiently inhibit cellular proliferation . In the present study , we show that OT increases invasiveness of human endometrial carcinoma ( O14777 ) cells , which are otherwise resistant to the growth-inhibiting effects of OT . Using pharmacological inhibitors , invasion assay , RNA interference , and immunofluorescence , we found that OT enhances the invasive properties of O14777 cells through up-regulation of P98170 ( P98170 ) , matrix-metalloproteinase 2 ( P08253 ) , and matrix-metalloproteinase 14 ( P50281 ) . In addition , we show that OT-mediated invasion is both cyclooxygenase 1 ( P23219 ) and cyclooxygenase-2 ( P35354 ) dependent via the phosphatidylinositol 3-kinase/AKT ( PIK3/AKT ) pathway . P35354 knockdown by shRNA resulted in P98170 down-regulation . We also show that OT receptor is overexpressed in grade I to III endometrial cancer . Taken together , our results describe for the first time a novel role for OT in endometrial cancer cell invasion . Prostaglandin-endoperoxide synthase genes P23219 and P35354 - novel modifiers of disease severity in cystic fibrosis patients . Cystic fibrosis ( CF ) is one of the most common autosomal recessive diseases among Caucasians caused by a mutation in the P13569 gene . However , the clinical outcome of CF pulmonary disease varies remarkably even in patients with the same P13569 genotype . This has led to a search for genetic modifiers located outside the P13569 gene . The aim of this study was to evaluate the effect of functional variants in prostaglandin-endoperoxide synthase genes ( P23219 and P35354 ) on the severity of lung disease in CF patients . To the best of our knowledge , it is the first time when analysis of P23219 and P35354 as potential CF modifiers is provided . The study included 94 CF patients homozygous for F508del mutation of P13569 . To compare their clinical condition , several parameters were recorded , e.g. a unique clinical score : disease severity status ( DSS ) . To analyse the effect of non- P13569 genetic polymorphisms on the clinical course of CF patients , the whole coding region of P23219 and selected P35354 polymorphisms were analysed . Statistical analysis of genotype-phenotype associations revealed a relationship between the heterozygosity status of identified polymorphisms and better lung function . These results mainly concern P35354 polymorphisms : -765G > C and 8473T > C . The P23219 and P35354 polymorphisms reducing P36551 protein levels had a positive effect on all analysed clinical parameters . This suggests an important role of these genes as protective modifiers of pulmonary disease in CF patients , due to inhibition of arachidonic acid conversion into prostaglandins , which probably reduces the inflammatory process . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . 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 . Application of Q99697 box proteins to evaluate G-protein selectivity in receptor-promoted signaling . Regulator of G-protein signaling ( Q99697 ) domains bind directly to GTP-bound Galpha subunits and accelerate their intrinsic GTPase activity by up to several thousandfold . The selectivity of Q99697 proteins for individual Galpha subunits has been illustrated . Thus , the expression of Q99697 proteins can be used to inhibit signaling pathways activated by specific G protein-coupled receptors ( GPCRs ) . This article describes the use of specific Q99697 domain constructs to discriminate among G(i/o) , Gq-and G(12/13)-mediated activation of phospholipase C ( P98160 ) isozymes in COS-7 cells . Overexpression of the N terminus of P25098 ( amino acids 45-178 ) or P98171 RhoGEF ( amino acids 1-240 ) elicited selective inhibition of Galphaq- or Galpha(12/13)-mediated signaling to P98160 activation , respectively . In contrast , P41220 overexpression was found to inhibit P98160 activation by both G(i/o)- and Gq-coupled GPCRs . P49798 exhibited dramatic receptor selectivity in its inhibitory actions ; of the G(i/o)- and Gq-coupled GPCRs tested ( Q92633 , Q9HBW0 , P47900 , Q99500 ) , only the Gq-coupled lysophosphatidic acid-activated Q9HBW0 receptor was found to be inhibited by P49798 overexpression . Purinergic Q15077 receptors induce Ca2+ and P13569 dependent Cl- secretion in mouse trachea . In airways Cl- secretion is activated and Na+ absorption is inhibited when P41231 receptors are stimulated by DB00171 or UTP . Both nucleotides are subject to degradation to ADP and UDP by ecto-nucleotidases . Here we show that these metabolites change electrolyte transport by stimulation of Q15077 receptors in mouse trachea . Immunohistochemistry confirmed luminal and basolateral expression of Q15077 receptors . In Ussing chamber experiments luminal ADP , UDP or the Q15077 receptor agonist INS48823 induced both transient and persistent increase in short circuit currents ( ISC ) . Activation of ISC was inhibited by the Q15077 receptor blocker PPADS . The transient response was inhibited by DIDS , whereas the persistent ISC was inhibited by glibenclamide and by the protein kinase A ( PKA ) blocker H-89 . Moreover , sustained activation of ISC by luminal UDP was inhibited by blocking basolateral K+ channels with 293B . Possible effects of diphosphates on P47900 or adenosine receptors were excluded by the inhibitors MRS2179 and 8- P21549 , respectively . Inhibition of amiloride sensitive Na+ absorption was only seen after blocking basolateral K+ channels with 293B . In contrast , Cl- secretion activated by basolateral ADP or UDP was only transient and was blocked by the sk4 K+ channel blocker clotrimazole . In summary , activation of luminal Q15077 receptors in the airways shifts electrolyte transport towards secretion by increasing intracellular Ca+ and activation of PKA . 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 genetic background of ulcer diseases induced by NSAID/aspirin ] . The association between peptic ulcer diseases and polymorphisms in various genes , including P25021 , P23219 , Q16552 . Q96PD4 , MIF and Nrf2 genes , are seen . P23219 has traditionally been regarded as a constitutively expressed enzyme that generates prostaglandins for gastrointestinal integrity . The effects of NSAID/aspirin on the gastric mucosal damage are caused by the inhibition of this enzyme . A T-1676C polymorphism ( rs1330344 ) was significantly associated with the development of peptic ulcer , especially gastric ulcer . In addition , rs1330344 was also significantly associated with the development of NSAID/aspirin-induced ulcer diseases . In conclusions , the assessment for genotype of P23219 gene promoter polymorphism , especially rs1330344 , may be useful for detecting the high risk group of developing NSAID/aspirin-induced ulcer diseases . 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 . 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 . 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 . [ Changes in nitric oxide , prostaglandins and myeloperoxidase activity in acrolein-induced cystitis in rats ] . To investigate the role of DB05875 ( sP ) , nitric oxide ( ON ) and prostaglandins ( PGs ) in acrolein ( P10323 ) -induced cystitis , we studied the changes induced by P10323 on bladder inducible nitric oxide synthase ( P35228 ) and mieloperoxidase ( P05164 ) activities , along with PGs and NO metabolites levels . Sprague-Dawley male rats received i.p . P10323 ( 5 mg/Kg ) plus one of the following treatments : Group 1 : saline 0.10 mL/100g i.p. ; Group 2 : Win-51.708 ( Q08050 ) 25 mg/Kg i.p. ; Group 3 : S-metilisothiourea ( MITU ) 35 mg/Kg i.p. ; Group 4 : DB00533 ( DB01656 ) 20 mg/Kg o.p. ; Group 5 : Meloxicam( P61006 ) 25 mg/Kg i.p. ; Group 6 : combination MITU+ P61006 . P10323 -induced mortality was partially prevented by Q08050 ( NK1 antagonist ) and MITU ( P35228 inhibitor ) . Animals that survived after 24h of P10323 exposure , had histological inflammatory changes in bladder along with increased P05164 activity . There was augmentation of nitrates+nitrites and of PGs . Q08050 did n't prevent any of these effects . DB01656 and P61006 ( P35354 inhibitors ) partially protected against bladder inflammation ; MITU pre-treatment was able to prevent these changes and those of NO metabolites levels . The MITU+ P61006 combination produced the highest protection against P10323 -induced damage . These results suggest that NO produced via P35228 and PGs produced by P23219 / P35354 , have an important role in the pathogenesis of cystitis induced by P10323 . P10323 could stimulate P35228 and P23219 / P35354 , producing lymphocyte migration and increases of NO and PGs . Nucleotide P47900 receptor regulates P01133 receptor mitogenic signaling and expression in epithelial cells . P00533 ( P00533 ) function is transregulated by a variety of stimuli , including agonists of certain G-protein-coupled receptors ( GPCRs ) . One of the most ubiquitous GPCRs is the P2Y(1) receptor ( P47900 , hereafter referred to as P2Y(1)R ) for extracellular nucleotides , mainly ADP . Here , we show in tumoral HeLa cells and normal P17948 epithelial cells that P2Y(1)R broadcasts mitogenic signals by transactivating the P00533 . The pathway involves PKC , Src and cell surface metalloproteases . Stimulation of P2Y(1)R for as little as 15-60 minutes triggers mitogenesis , mirroring the half-life of extracellular ADP . Apyrase degradation of extracellular nucleotides and drug inhibition of P2Y(1)R , both reduced basal cell proliferation of HeLa and P17948 cells , but not MDCK cells , which do not express P2Y(1)R . Thus , cell-released nucleotides constitute strong mitogenic stimuli , which act via P2Y(1)R . Strikingly , MDCK cells ectopically expressing P2Y(1)R display a highly proliferative phenotype that depends on P00533 activity associated with an increased level of P00533 , thus disclosing a novel aspect of GPCR-mediated regulation of P00533 function . These results highlight a role of P2Y(1)R in P00533 -dependent epithelial cell proliferation . P2Y(1)R could potentially mediate both trophic stimuli of basally released nucleotides and first-line mitogenic stimulation upon tissue damage . It could also contribute to carcinogenesis and serve as target for antitumor therapies .

MH_train_1119

MH_train_1119

MH_train_1119

interacts_with DB01268?

multiple_choice

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 . 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 . 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 . Differentiation of human embryonic stem cells into cells with corneal keratocyte phenotype . Corneal transparency depends on a unique extracellular matrix secreted by stromal keratocytes , mesenchymal cells of neural crest lineage . Derivation of keratocytes from human embryonic stem ( hES ) cells could elucidate the keratocyte developmental pathway and open a potential for cell-based therapy for corneal blindness . This study seeks to identify conditions inducing differentiation of pluripotent hES cells to the keratocyte lineage . Neural differentiation of hES cell line WA01(H1) was induced by co-culture with mouse PA6 fibroblasts . After 6 days of co-culture , hES cells expressing cell-surface P08138 protein ( CD271 , p75NTR ) were isolated by immunoaffinity adsorption , and cultured as a monolayer for one week . Keratocyte phenotype was induced by substratum-independent pellet culture in serum-free medium containing ascorbate . Gene expression , examined by quantitative RT-PCR , found hES cells co-cultured with PA6 cells for 6 days to upregulate expression of neural crest genes including P08138 , O95863 , Q16288 , P48436 , and P28360 . Isolated P08138 -expressing cells were free of PA6 feeder cells . After expansion as a monolayer , mRNAs typifying adult stromal stem cells were detected , including P35226 , P10721 , P48681 , P46531 , and Q9NPC8 . When these cells were cultured as substratum-free pellets keratocyte markers P29972 , Q8NFL0 , PTDGS , and P30838 were upregulated . mRNA for keratocan ( O60938 ) , a cornea-specific proteoglycan , was upregulated more than 10,000 fold . Culture medium from pellets contained high molecular weight keratocan modified with keratan sulfate , a unique molecular component of corneal stroma . These results show hES cells can be induced to differentiate into keratocytes in vitro . Pluripotent stem cells , therefore , may provide a renewable source of material for development of treatment of corneal stromal opacities . DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . 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 . Combined Effects of Q07869 γ Agonists and Epidermal Growth Factor Receptor Inhibitors in Human Proximal Tubule Cells . We aimed to determine whether epidermal growth factor receptor ( P00533 ) inhibition , in addition to a peroxisome proliferator-activated receptor gamma ( Q07869 γ ) agonist , prevents high-glucose-induced proximal tubular fibrosis , inflammation , and sodium and water retention in human proximal tubule cells exposed to normal glucose ; high glucose ; high glucose with the Q07869 γ agonist pioglitazone or with the P- P00533 inhibitor , gefitinib ; or high glucose with both pioglitazone and gefitinib . We have shown that high glucose increases AP-1 and NF κ B binding activity , downstream phosphorylation of P00533 and Erk1/2 , and fibronectin and collagen IV expression . Pioglitazone reversed these effects but upregulated P48764 and P29972 expression . Gefitinib inhibited high glucose induced fibronectin and collagen IV , and P00533 and Erk1/2 phosphorylation and reversed pioglitazone-induced increases in P48764 and P29972 expression . Our data suggests that combination of an P00533 inhibitor and a Q07869 γ agonist mitigates high-glucose-induced fibrosis and inflammation and reverses the upregulation of transporters and channels involved in sodium and water retention in human proximal tubule cells . Hence P00533 blockade may hold promise , not only in limiting tubulointerstitial pathology in diabetic nephropathy , but also in limiting the sodium and water retention observed in patients with diabetes and exacerbated by Q07869 γ agonists . 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 . DB04868 : a phenylamino-pyrimidine derivative with activity against P11274 - P00519 , P10721 and P09619 kinases . The P11274 - P00519 kinase inhibitor imatinib mesylate is currently the standard therapy for patients with chronic myeloid leukemia ( CML ) . However , mutations within the P00519 kinase domain interfering with drug binding have been identified as the main mechanism of resistance to imatinib . Multiple distinct P11274 - P00519 kinase mutant isoforms conferring varying degrees of resistance to tyrosine kinase inhibitors have been reported . DB04868 is a tyrosine kinase inhibitor 30-fold more potent than imatinib against P11274 - P00519 kinase . DB04868 is active against a wide range of imatinib-resistant P11274 - P00519 mutant isoforms , except for T315I . Results from Phase II studies of nilotinib for patients with CML after failure or intolerance to imatinib therapy have shown a favorable toxicity profile and confirmed the high efficacy of nilotinib in this setting . Studies addressing the activity of nilotinib in newly-diagnosed patients with CML are underway . Furthermore , nilotinib is a potent inhibitor of P10721 and P09619 kinases . Here , we review the preclinical development of nilotinib and the activity of this agent in patients with CML and in tumors driven by P10721 and/or P09619 mutant kinases , such as gastrointestinal stromal tumors and some forms of clonal hypereosinophilia . DB01268 induces P60484 expression and inhibits P09619 signaling and migration of medulloblastoma cells . We previously showed that inhibition of the platelet-derived growth factor receptor ( P09619 ) blocks the survival and migration of medulloblastoma cells . Identification of in vitro P09619 -targeting pharmacologic agents that are suitable for preclinical testing in medulloblastoma models in vivo will be critical for efficiently translating these agents to clinical investigation in children with medulloblastoma . In this study , we investigated whether the multi-tyrosine kinase inhibitor sunitinib , effectively inhibits P09619 signaling required for medulloblastoma cell migration . Daoy and D556 human medulloblastoma cells pre-treated for 1 h with 0.2 μM sunitinib demonstrated induction of P60484 expression and significant inhibition of P09619 signaling activity and transactivation of P00533 , in a DB01367 -independent manner , in response to DB00102 stimulation . DB01268 pre-treatment markedly reduced medulloblastoma cell migration in response to both DB00102 and 10 % serum at 4 and 24 h after treatment . Pre-treatment with sunitinib for 1 h also resulted in detachment and decreased viability of D556 , but not Daoy , cells and only after 48 h following treatment . However , sunitinib did not induce apoptosis in either cell line at any time point , indicating that the anti-migratory effects of sunitinib were not due to impeding cell survival . DB01268 similarly inhibited P09619 signaling and migration of primary murine Smo/Smo medulloblastoma cells , suggesting that the Smo/Smo mouse is an appropriate model for preclinical testing of sunitinib . These results indicate that sunitinib may be an important pharmacologic agent for the treatment of invasive medulloblastoma , particularly given evidence of its ability to cross the blood-brain barrier to target tumor cells , and thus warrants further in vivo testing for confirmation of efficacy . Identification of multiple rare variants associated with a disease . Identifying rare variants that are responsible for complex disease has been promoted by advances in sequencing technologies . However , statistical methods that can handle the vast amount of data generated and that can interpret the complicated relationship between disease and these variants have lagged . We apply a zero-inflated Poisson regression model to take into account the excess of zeros caused by the extremely low frequency of the 24,487 exonic variants in the Genetic Analysis Workshop 17 data . We grouped the 697 subjects in the data set as Europeans , Asians , and Africans based on principal components analysis and found the total number of rare variants per gene for each individual . We then analyzed these collapsed variants based on the assumption that rare variants are enriched in a group of people affected by a disease compared to a group of unaffected people . We also tested the hypothesis with quantitative traits Q1 , Q2 , and Q4 . Analyses performed on the combined 697 individuals and on each ethnic group yielded different results . For the combined population analysis , we found that P22309 , which was not part of the simulation model , was associated with disease liability and that P17948 , which was a causal locus in the simulation model , was associated with Q1 . Of the causal loci in the simulation models , P17948 and P35968 were associated with Q1 and O95497 was correlated with Q2 . No significant genes were associated with Q4 . These results show the feasibility and capability of our new statistical model to detect multiple rare variants influencing disease risk . Long-term response and postsurgical complete remissions after treatment with sunitinib malate , an oral multitargeted receptor tyrosine kinase inhibitor , in patients with metastatic renal cell carcinoma . Receptor tyrosine kinase ( RTK ) inhibitors have revolutionized the treatment of metastatic renal cell carcinoma ( mRCC ) and significantly extended survival in these patients . DB01268 is an oral multitargeted inhibitor of vascular endothelial growth factor receptors ( VEGFRs-1 , -2 , and -3 ) , platelet-derived growth factor receptors ( PDGFRs-α and -β ) , stem-cell factor receptor ( P10721 ) , P07333 -like tyrosine kinase 3 ( P36888 ) , colony-stimulating factor 1 receptor ( P07333 ) , and glial cell line-derived neurotrophic factor receptor ( REarranged during Transfection ; P07949 ) . DB01268 is approved multinationally for the treatment of advanced RCC , and is considered the reference standard of care for first-line treatment . In clinical trials , sunitinib has been associated with a consistent , distinct profile of adverse events . Here we describe three cases that show that it is possible to manage adverse events occurring during sunitinib therapy , and thus allow patients with mRCC to receive an effective dose of sunitinib in order to achieve long-term disease control . These cases also show that surgical resection , performed whenever possible , can help to improve control of metastatic disease and so avoid the unnecessary toxicity and high costs of prolonged antiangiogenic therapy . 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 . P36888 K663Q is a novel AML-associated oncogenic kinase : Determination of biochemical properties and sensitivity to DB01268 ( SU11248 ) . Somatic mutations of P36888 resulting in constitutive kinase activation are the most common acquired genomic abnormality found in acute myeloid leukemia ( AML ) . The majority of these mutations are internal tandem duplications ( ITD ) of the juxtamembrane region ( JM ) . In addition , a minority of cases of AML are associated with mutation of the P36888 activation loop ( AL ) , typically involving codons D835 and/or I836 . We hypothesized that other novel mutations of P36888 could also contribute to leukemogenesis . We genotyped 109 cases of AML and identified two novel gain-of-function mutations . The first mutation , N841 H , is similar to previously described mutations involving amino-acid substitutions of codon 841 . The other novel mutation , P36888 K663Q , is the first AML-associated gain-of-function mutation located outside the JM and AL domains . Of note , this mutation was potently inhibited by DB01268 ( SU11248 ) , a previously described P36888 kinase inhibitor . DB01268 reduced the proliferation and induced apoptosis of transformed Ba/ P13726 cells expressing P36888 K663Q . The potency of DB01268 against P36888 K663Q was similar to its potency against P36888 ITD mutations . We conclude that P36888 mutations in AML can involve novel regions of the P04183 . Future studies are needed to define the incidence and prognostic significance of P36888 mutations outside the well-established JM and AL regions . 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 . 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 . 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 . 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 . DB01268 suppresses tumor growth and metastases in a highly metastatic mouse mammary cancer model . BACKGROUND : DB01268 is an inhibitor that blocks tyrosine phosphorylation ( p- DB00135 ) of receptors including vascular endothelial growth factor receptor ( VEGFR ) and platelet-derived growth factor receptor . DB01268 suppresses angiogenesis and cell proliferation and is an effective treatment for renal cell carcinoma and gastrointestinal stromal tumors . In the present study , we examined the antitumor and antimetastatic activities of sunitinib in mouse metastatic mammary cancer . MATERIALS AND METHODS : Mammary tumors induced by inoculation of BJMC3879 cells into mice were treated with sunitinib using mini-osmotic pumps . At 1 week and 7 weeks after initiation of drug administration , cancer tissue was removed and carried out histopathological and immunohistochemical examination . RESULTS : Tumor growth , as well as metastasis to the lungs and other organs , was significantly inhibited in sunitinib-treated mice . Cell death areas in mammary carcinomas were much larger in the sunitinib-treated groups than in the control group . In addition , sunitinib induced necrotic cell death rather than apoptosis . Although microvessel density was significantly lower in the sunitinib-treated mammary tumors , numbers of metastases to lymph nodes and the number of lymphatic vessels in the mammary tumors were not significantly different among groups . Cell proliferation , as assessed by BrdU-labeling indices , was significantly lower in mammary carcinomas of sunitinib-treated mice . The amounts of p- P35968 and p- DB00135 , as determined by immunohistochemistry , were greatly reduced in sunitinib-treated mice . CONCLUSION : In a mouse model of mammary cancer , sunitinib inhibited tumor growth and metastasis ( with the exception of lymph node metastasis ) , angiogenesis , and cell proliferation possibly due to reduced levels of p- P35968 and p- DB00135 . O95760 augments DB05875 -induced P15692 secretion from human mast cells and is increased in psoriatic skin . The peptide DB05875 ( SP ) has been implicated in inflammatory conditions , such as psoriasis , where mast cells and P15692 are increased . A relationship between SP and P15692 has not been well studied , nor has any interaction with the proinflammatory cytokines , especially O95760 . Here we report that SP ( 0.1-10 microM ) induces gene expression and secretion of P15692 from human LAD2 mast cells and human umbilical core blood-derived cultured mast cells ( hCBMCs ) . This effect is significantly increased by coadministration of O95760 ( 5-100 ng/mL ) in both cell types . The effect of SP on P15692 release is inhibited by treatment with the P25103 antagonist 733,060 . SP rapidly increases cytosolic calcium , and so does O95760 to a smaller extent ; the addition of O95760 augments the calcium increase . SP-induced P15692 production involves calcium-dependent PKC isoforms , as well as the P29323 and JNK MAPKs . Gene expression of O95760 and histidine decarboxylase ( HDC ) , an indicator of mast cell presence/activation , is significantly increased in affected and unaffected ( at least 15 cm away from the lesion ) psoriatic skin , as compared with normal control skin . Immunohistochemistry indicates that O95760 is associated with endothelial cells in both the unaffected and affected sites , but is stronger and also associated with immune cells in the affected site . These results imply that functional interactions among SP , O95760 , and mast cells leading to P15692 release contribute to inflammatory conditions , such as the psoriasis , a nonallergic hyperproliferative skin inflammatory disorder with a neurogenic component . The role of HIV-related chemokine receptors and chemokines in human erythropoiesis in vitro . In order to better define the role of HIV-related chemokines in human erythropoiesis we studied : A ) the expression of chemokine receptors , both on human P28906 (+) cells which include erythroid progenitors and on more mature erythroid cells ; B ) the functionality of these receptors by calcium flux , chemotaxis assay and phosphorylation of mitogen-activated protein kinases ( MAPK ) Q8NFH3 /44 ( P27361 / P28482 ) and AKT , and finally C ) the influence of chemokines on BFU-E formation . We found that HIV-related chemokine receptor P61073 , but not P51681 , is detectable on human P28906 (+) BFU-E cells . P61073 surface expression decreased during erythroid maturation , although P61073 mRNA was still present in cells isolated from differentiated erythroid colonies . P48061 , a P61073 ligand , induced calcium flux and phosphorylation of MAPK ( Q8NFH3 /44 ) and AKT in P28906 (+) P10721 (+) bone marrow mononuclear cells which contain BFU-E , as well as chemotactic activity of both human P28906 (+) BFU-E progenitors and erythroid cells isolated from day 2-6 BFU-E colonies . Responsiveness to P48061 decreased when the cells differentiated to the point of surface expression of the erythroid-specific marker P02724 . In contrast , the P51681 ligands ( macrophage inflammatory protein-1alpha [ MIP-1alpha ] , MIP-1beta , and RANTES ) did not activate calcium flux , MAPK and AKT phosphorylation or chemotaxis of P28906 (+) P10721 (+) cells or cells isolated from the BFU-E colonies . Interestingly , none of the chemokines tested in this study had any effect on BFU-E colony formation . In conclusion , only P61073 is functional , and its specific ligand P48061 may therefore play an important role in the homing and/or retention of early erythroid precursors in the bone marrow environment . P01308 /FGF-binding ciliary membrane glycoprotein from Tetrahymena . Triton X-100 extracted ciliary membrane protein from isolated cilia , prepared from the protozoon Tetrahymena thermophila , were fractionated by affinity chromatography on columns with covalently bound fibroblast growth factor ( FGF ) , insulin , or concanavalin A ( ConA ) , respectively . The eluted proteins were further analyzed by electrophoresis on sodium dodecyl sulfate polyacrylamide gels , isoelectric focusing , and by immunoblotting techniques using antibodies against the FGF receptor , platetelet derived growth factor ( PDGF ) receptor alpha-subunit , and insulin receptor beta-subunit . The particular antibodies were chosen because the peptides PDGF , FGF , insulin , and ConA are chemoattractants in this organism and corresponding binding ( receptor ) proteins could be expected to be identified . A 66 kDa protein fraction was eluted from the FGF-MiniLeak agarose , insulin-MiniLeak agarose and ConA sepharose . This fraction responded in Western immunoblots to an antibody against the beta-subunit of the human insulin receptor , to an antibody against the PDGF receptor ( P09619 ) and also to an antibody against the bovine FGF receptor ( FGFR ) that is known , in other systems , to inhibit FGF binding to its receptor . When analyzed by SDS-PAGE and stained with Coomassie blue the 66 kDa fraction appeared as a single component . However , in some experiments it appeared more heterogeneous when stained with silver indicating the presence of minor components that may be a procedural artifact or isoforms of the same glycoprotein . The 66 kDa protein(s) migrated in isoelectric focusing with a pI of 7.4 . The results are discussed in terms of the possible role of the 66 kDa glycoprotein as a protein involved in peptide-mediated cell signalling . Application of HapMap data to the evaluation of 8 candidate genes for pediatric slow transit constipation . BACKGROUND : Slow transit constipation ( P52823 ) affects up to 3 % of all children and is an increasingly recognized cause of chronic constipation in children . We conducted a pilot study to investigate whether genes encoding neurotransmitters ( P20366 , Q9UHF0 , P01282 , NOS1 ) and receptors ( P25103 , P21452 , P29371 , P10721 ) could be responsible for P52823 . METHODS : One hundred seventeen tag single nucleotide polymorphisms ( SNPs ) , distributed among the candidate genes , were selected from HapMap data and genotyped using Sequenom ( San Diego , CA ) technology in 35 affected families . Evaluation of association was performed by transmission disequilibrium test and multilocus analysis . RESULTS : Five SNPs ( rs3771863 , rs4580655 , rs11722288 , rs4563545 , and rs3782221 ) in the P25103 , P29371 , P10721 , and NOS1 genes were found to be potentially associated with P52823 , although the significance of these results does not withstand correction for multiple testing . CONCLUSIONS : Our data indicate that 5 SNPs in the NOS1 , P25103 , P29371 , and P10721 genes could be involved in P52823 , especially rs3771863 in intron 1 of P25103 , which showed the highest association . Prolonged high fat/alcohol exposure increases Q9HBA0 and its functional responses in pancreatic stellate cells . The present study investigated transient receptor potential vanilloid type 4 ( Q9HBA0 ) ion channels in pancreatic stellate cells ( PSCs ) isolated from rats with high-fat and alcohol diet ( HFA ) -induced chronic pancreatitis . Q9HBA0 is a calcium-permeable nonselective ion channel responsive to osmotic changes , alcohol metabolites arachidonic acid , anandamide , their derivatives , and injury-related lipid mediators . Male Lewis rats were fed HFA for 6-8 wk before isolation and primary culture of PSCs . Control PSCs were harvested from rats fed standard chow . Immunoreactivity for cytoskeletal protein activation product α-smooth muscle actin ( α-SMA ) and platelet-derived growth factor receptor-β subunit ( P09619 -β ) characterized the cells as PSCs . Q9HBA0 expression increased in PSCs of HFA-fed rats and control cultures after alcohol treatment ( 50 mM ) . Cell responses to activation of inducible Q9HBA0 were assessed with live cell calcium imaging . Threefold increased and sustained intracellular calcium mobilization responses occurred in 70 % of pancreatic stellate cells from HFA-fed rats in response to Q9HBA0 activators arachidonic acid , lipid second messenger , phorbol ester 4 α-phorbol 12,13-didecanoate ( 4αPDD ) , and 50 % hypoosmotic media compared with relatively unresponsive PSCs from control rats . Activation responses were attenuated by nonselective TRPV channel blocker ruthenium red . P01375 -α ( P01375 -α , 1 ng/ml , 16 h ) increased responses to 4αPDD in control PSCs . These findings implicate Q9HBA0 -mediated calcium responses inducible after HFA exposure and inflammation in reactive responses of activated PSCs that impair pancreatic function , such as responsiveness to cytokines and the deposition of collagen fibrosis that precipitates ductal blockage and pain . 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 . Methylation analysis of several tumour suppressor genes shows a low frequency of methylation of CDKN2A and P10826 in uveal melanomas . We have investigated the frequency of methylation of several tumour suppressor genes in uveal melanoma . As the loss of one copy of chromosome 3 ( monosomy 3 ) , which is found in about half of these tumours , is tightly associated with metastatic disease , a special emphasis was laid on genes located on this chromosome , including the fragile histidine triad ( P49789 ) , von Hippel-Lindau ( P40337 ) , beta-catenin ( P35222 ) , activated leukocyte cell adhesion molecule ( Q13740 ) and retinoic acid receptor-beta2 ( P10826 ) genes . In addition , the methylation patterns of the CpG-rich regions 5' of the P12830 ( CDH1 ) , p16/cyclin-dependent kinase inhibitor 2 A ( CDKN2A ) and retinoblastoma ( P06400 ) genes were analysed by bisulphite genomic sequencing or methylation-specific PCR ( MSP ) . Furthermore , the P63162 and D15S63 loci , which are located in the imprinted region of chromosome 15 , were included in the study . Aberrant methylation was detected in nine of 40 tumours analysed : The imprinted P63162 and D15S63 loci were hypermethylated in three tumours , all of which retained both copies of chromosome 3 . Methylated P10826 alleles were detected in three tumours , whereas in three other tumours CDKN2A was found to be methylated . As we did not find P10826 and CDKN2A preferentially methylated in tumours with monosomy 3 , which is a significant predictor of metastatic disease , we suggest that these genes may play a causative role in the formation of uveal melanoma but not in the development of metastases . DB01268 deregulates tumor adaptation to hypoxia by inhibiting HIF-1alpha synthesis in HT-29 colon cancer cells . DB01268 ( SU11248 , Sutent ) is a class III/V receptor tyrosine kinase ( RTK ) inhibitor that exhibits potent anti-angiogenic and anticancer activities . Preclinical studies demonstrated that the sunitinib effects are attributed to inhibition of VEGFR and P09619 phosphorylation . However , even in colon cancer cells lacking sunitinib-targeted RTKs , sunitinib effectively inhibits tumor growth in a xenograft model , and this raises a question about the mechanism underlying the in vivo anticancer action of sunitinib . Since hypoxia is a critical microenvironment that tumors face , we addressed the possibility that sunitinib deregulates tumor adaptation to hypoxia . First we found that sunitinib limits the colony growth of HT-29 , which is a colon adenocarcinoma cell line lacking the RTKs , and that HIF-1alpha in the colonies is decreased by sunitinib . In cultured HT-29 cells , sunitinib suppressed HIF-1alpha under hypoxic conditions . Moreover , sunitinib repressed the activity of HIF-1alpha and subsequently decreased the expressions of Q9BYW2 downstream genes . Mechanistically , sunitinib blocked the 5'-UTR-dependent translation of HIF-1alpha . The HIF-1alpha suppression by sunitinib was also reproduced in a P40337 -null renal cell carcinoma cell line , where HIF-1alpha is not degradable . In conclusion , the sunitinib inhibition of Q9BYW2 signaling could restrain tumor progression in hypoxic regions , which may contribute to anticancer effect of sunitinib . 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 . 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 . Tolerability and pharmacokinetic profile of a sunitinib powder formulation in pediatric patients with refractory solid tumors : a Children 's Oncology Group study . PURPOSE : DB01268 is an oral tyrosine kinase inhibitor of P15692 , PDGF , c- P10721 , and flt-3 receptors . A pediatric phase I study of sunitinib capsules identified the maximum tolerated dose as 15 mg/m(2)/day . This study was conducted to evaluate sunitinib given as a powder formulation . METHODS : DB01268 15 mg/m(2) was administered orally daily for 4 weeks on/2 weeks off to patients < 21 years old with refractory solid tumors . DB01268 capsules were opened , and the powder sprinkled onto applesauce or yogurt . Plasma levels of sunitinib and an active metabolite , SU12662 , were measured , and pharmacokinetic parameters were estimated . RESULTS : 12 patients , median age 13 ( range 4-21 ) years , were treated . The most common first-cycle toxicities were leucopenia ( n = 6 ) , fatigue ( n = 5 ) , neutropenia ( n = 4 ) , and hypertension ( n = 4 ) . Three patients had dose-limiting toxicities ( DLTs ) in cycle 1 ( dizziness/back pain , hand-foot syndrome , and intratumoral hemorrhage/hypoxia ) . A median peak plasma sunitinib concentration of 21 ( range 6-36 ) ng/ml was reached at a median of 4 ( range 4-8 ) h after the first dose . The median exposure ( AUC(0-48) ) was 585 ( range 196-1,059 ) h ng/l . The median half-life was 23 ( range 13-36 ) h . The median trough concentration measured before day 14 dosing was 32 ( range 12-58 ) ng/ml . CONCLUSIONS : The pharmacokinetic profile of sunitinib appears similar between a powder formulation and published data using capsules . The powder formulation allows patients unable to swallow capsules to receive sunitinib . 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 . DB01268 as a second-line therapy for advanced GISTs after failure of imatinib : relationship between efficacy and tumor genotype in Korean patients . BACKGROUND : To assess the efficacy and safety of sunitinib with regards to primary genotypes of tumor in Korean patients with advanced gastrointestinal stromal tumors ( GISTs ) who failed an initial therapy of imatinib . METHODS : Clinical data were collected from 88 consecutive patients with metastatic/unresectable GISTs treated with sunitinib at the Asan Medical Center . RESULTS : The median time-to-progression ( TTP ) and overall survival ( OS ) times were 7.1 months and 17.6 months , respectively . Of the 74 patients tested for P10721 ( exons 9 , 11 , 13 , 17 ) and P16234 ( exons 12 and 18 ) , patients with P10721 exon 9 mutant GIST ( n = 11 , 14.9 % ) showed numerically better clinical benefit ( objective response or stable disease ≥ 24 weeks ) rate ( 63.6 % vs 46.8 % , p = 0.504 ) and TTP ( median 13.6 mo vs 6.9 mo , p = 0.631 ) than those with P10721 exon 11 mutant GIST ( n = 47 , 63.5 % ) . The most common grade 3/4 adverse events included neutropenia ( 34.1 % ) , thrombocytopenia ( 33.0 % ) and hand-foot skin reaction ( 25.0 % ) . CONCLUSIONS : DB01268 is an effective and safe second-line therapy for Korean patients with advanced GIST . The superior efficacy of sunitinib against GISTs with P10721 exon 9 mutations appears to be similar in Korean patients to Western experience although statistical significance was not secured . 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 . Biological and immunological studies of bovine hypothalamic DB05394 . P06850 B ( CRF-B ) is a peptide(s) isolated from bovine hypothalamic extracts by Sephadex G-100 chromatography on the basis of its ability to stimulate secretion of adrenocorticotropin ( DB01285 ) in vitro and in vivo . It is similar in molecular size to the 41-residue ovine CRF ( oCRF ) or rat CRF ( rCRF ) recently elucidated and appears to be their bovine counterpart . Immunoreactivity of CRF-B was examined in homologous radioimmunoassays ( RIAs ) for oCRF or rCRF , using several anti-oCRF and anti-rCRF antibodies . CRF-B cross-reacted well with anti-oCRF antibodies but poorly with anti-rCRF antibodies . Purification of CRF-B with preparative isoelectric focusing yielded four CRF peaks , B-1 ( pH 4.7 ) , B-2 ( pH 5.5 ) , B-3 ( pH 6.3 ) , and B-4 ( pH 7.0 ) , which accounted for 16 , 30 , 46 , and 8 % of the total immunoreactivity , respectively . CRF B-2 , B-3 , and B-4 showed both immunological activity and biological activity in vitro ( cell culture assay ) and in vivo ( Arimura assay ) , whereas CRF B-1 showed only immunoreactivity . Their relative bioactivity/immunoreactivity ratios were 0 ( B-1 ) , 1 ( B-2 ) , 1 ( B-3 ) , and 3 ( B-4 ) . All of these CRF-B subtypes exhibited RIA displacement curves parallel to that for the oCRF standard and coeluted with oCRF on Sephadex G-100 chromatography , which suggests that their molecular modifications are relatively minor . Platelet-derived growth factor ( PDGF ) -BB inhibits AMPA receptor-mediated synaptic transmission via PDGF receptor-beta in murine nucleus tractus solitarius . Although platelet-derived growth factor ( PDGF ) -BB activates PDGF receptor-beta ( P09619 ) and , in turn , inhibits the glutamate N-methyl-D-aspartate ( DB01221 ) receptor function , whether DB00102 modulates the CNS function mediated by another glutamate receptors , alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ( AMPA ) receptors , remains poorly understood . Here we now report the inhibitory effect of DB00102 on the AMPA receptor function in the nucleus tractus solitarius ( P30990 ) by using slice patch-clamp techniques . Excitatory postsynaptic currents ( EPSCs ) were evoked by electrical stimulation of the tractus solitarius in mouse P30990 second-order neurons . EPSCs were nearly completely eliminated by CNQX but not by MK-801 , implying mediation through non- DB01221 receptors . DB00102 significantly decreased the amplitude of EPSCs without affecting the mean decay time constant . This inhibitory effect was transient and reversible after removing DB00102 . Furthermore , DB00102 significantly reduced the amplitude of AMPA-induced currents in P30990 neurons , which showed that DB00102 could suppress the AMPA receptor-mediated excitatory input via the postsynaptic mechanism . The inhibitory effect of DB00102 on EPSCs was not observed in mutant mice with conditional deletion of the P09619 gene in neurons . Together , these studies suggest that the PDGF-B/ P09619 axis inhibits the AMPA receptor-mediated synaptic transmission that comprises the major part of the primary afferent to the P30990 second-order neuron . The detected inhibitory action may be involved in the CNS regulation of the respiratory response . Second line therapies for the treatment of gastrointestinal stromal tumor . PURPOSE OF REVIEW : Most gastrointestinal stromal tumors eventually acquire resistance to imatinib mesylate . This review focuses on recent progress on management of patients whose disease progresses on the standard dose of imatinib . RECENT FINDINGS : Approximately 30 % of patients failing standard-dose imatinib achieve disease stabilization with high-dose imatinib , but objective responses are few and the clinical benefit usually short-lived . Patients receiving enzyme-inducing drugs may need high imatinib doses to achieve therapeutic blood concentrations . Surgical excision of a single growing metastasis leads to a median progression-free survival time of 7-11 months . DB01268 malate is effective following imatinib failure . The median time to disease progression is approximately 6 months with sunitinib therapy versus 6 weeks with placebo following discontinuation of imatinib , but few ( 5 % ) patients achieve objective response . Patients with gastrointestinal stromal tumor with P10721 exon 9 mutation may benefit more from sunitinib than those with exon 11 mutation . DB01268 frequently causes abnormal thyroid function . SUMMARY : DB01268 is now the approved second line therapy following imatinib failure and for patients intolerant to imatinib . The clinical benefit is only moderate , and thyroid function monitoring is required . Several investigational agents are being evaluated for imatinib-resistant gastrointestinal stromal tumor . Palliative procedures , such as hepatic arterial embolization , also require study . 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 . Pharmacologic modulation of niche accessibility via tyrosine kinase inhibition enhances marrow and thymic engraftment after hematopoietic stem cell transplantation . Essential survival signals within hematopoietic stem cell ( P19526 ) and thymic niches are mediated by receptor tyrosine kinases , which can be reversibly inhibited using clinically available drugs . We studied whether sunitinib , a multityrosine kinase inhibitor that inhibits P10721 , enhances engraftment after bone marrow transplantation ( BMT ) in mice . DB01268 diminished hematopoietic progenitor cell numbers , and sunitinib enhanced marrow , peripheral myeloid , and lymphoid engraftment after BMT in Rag1(-/-) mice . DB01268 augmented P19526 engraftment because recipients displayed increased myeloid and lymphoid engraftment and because sunitinib-treated recipients of purified HSCs showed enhanced engraftment of secondary hosts . However , sunitinib preferentially augmented T-cell engraftment with lesser effects on myeloid and P19526 engraftment . Consistent with this , sunitinib preferentially depleted the early thymic progenitor subset in the thymus . DB01268 did not increase engraftment in mice with deficient P10721 signaling , and the pattern of more potent effects on T cell compared with P19526 engraftment observed in sunitinib-treated hosts was also observed after BMT into P10721 (W/Wv) mice . These results implicate P10721 as a critical modulator of thymic niches . We conclude that transient , pharmacologic inhibition of P10721 enhances accessibility of marrow and thymic niches , and provides a novel , noncytotoxic approach to accomplish engraftment after stem cell transplantation . New perspectives : role of sunitinib 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 gastrointestinal stromal tumour ( GIST ) . DB01268 is able to inhibit RTKs such as receptors for platelet-derived growth factor ( PDGF-R alpha and beta ) and for vascular endothelial growth factor ( VEGFRs ) . It is able to inhibit P10721 receptor , colony stimulating factor type 1 receptor ( P04141 - 1R ) , 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 sunitinib targets play important roles in growth and survival of human breast cancer ( BC ) . The " rationale " of sunitinib 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 sunitinib in metastatic BC . Disease stabilization of progressive olfactory neuroblastoma ( esthesioneuroblastoma ) under treatment with sunitinib mesylate . Olfactory neuroblastoma ( esthesioneuroblastoma ) is a rare neoplasm of the olfactory epithelium in the upper nasal cavity . Here , we report the case of a 69-year-old man who presented with massive progression of a metastatic esthesioneuroblastoma after endonasal resection , functional neck dissection , and radiotherapy of local and distant tumor relapses . After exhaustion of all conventional therapeutic options , we initiated treatment with the oral multityrosinekinase inhibitor sunitinib mesylate . Using this drug , significant improvement of clinical symptoms , disease stabilization , and recovery from Karnofsky index of 40 % to 70 % could be achieved in the absence of significant adverse drug effects . The patient died 15 months after initiation of sunitinib therapy due to complications of a traumatic femoral neck fracture without evidence of tumor progression . Immunohistochemical analysis of tumor tissue specimens obtained at initial surgery revealed ample expression of platelet-derived growth factor receptor ( P09619 ) -b on stromal and endothelial cells . DB01268 should be considered for palliative therapy of advanced esthesioneuroblastoma . Blocking autocrine P15692 signaling by sunitinib , an anti-cancer drug , promotes embryonic stem cell self-renewal and somatic cell reprogramming . Maintaining the self-renewal of embryonic stem cells ( ESCs ) could be achieved by activating the extrinsic signaling , i.e. , the use of leukemia inhibitory factor ( P15018 ) , or blocking the intrinsic differentiation pathways , i.e. , the use of GSK3 and MEK inhibitors ( 2i ) . Here we found that even in medium supplemented with P15018 , mESCs still tend to differentiate toward meso-endoderm lineages after long-term culture and the culture spontaneously secretes vascular endothelial growth factors ( VEGFs ) . Blocking P15692 signaling with sunitinib , an anti-cancer drug and a receptor tyrosine kinase ( RTK ) inhibitor mainly targeting P15692 receptors ( VEGFRs ) , is capable of maintaining the mESCs in the undifferentiated state without the need for feeder cells or P15018 . DB01268 facilitates the derivation of mESCs from blastocysts , and the mESCs maintained in sunitinib-containing medium remain pluripotent and are able to contribute to chimeric mice . DB01268 also promotes iPSC generation from MEFs with only Oct4 . Knocking down P35968 or blocking it with neutralizing antibody mimicks the effect of sunitinib , indicating that blocking P15692 /VEGFR signaling is indeed beneficial to the self-renewal of mESCs . We also found that hypoxia-inducible factor alpha ( HIF1α ) and endoplasmic reticulum ( ER ) stress are involved in the production of P15692 in mESCs . Blocking both pathways inhibits the expression of P15692 and prevents spontaneous differentiation of mESCs . Interestingly , P15018 may also exert its effect by downregulating HIF1α and ER stress pathways and subsequent P15692 expression . These results indicate the existence of an intrinsic differentiation pathway in mESCs by activating the autocrine P15692 signaling . Blocking P15692 signaling with sunitinib or other small molecules help to maintain the mESCs in the ground state of pluripotency . 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 . Stable expression of a neuronal dopaminergic progenitor phenotype in cell lines derived from human amniotic fluid cells . Cells from human amniotic fluid derived from the fetus are considered a source of multipotent cells . Their properties have not been fully exploited , partially because unlike other embryonic sources such as embryonic stem ( ES ) cells , cell lines from amniocentesis samples have not been generated . We have established and characterized the properties of eight individual cell lines . Flow cytometry using several cell surface markers showed that all cell lines generated consisted of homogeneous populations that lack HLAII antigenicity . Using a combination of immunocytochemistry , Western blotting , and RT-PCR , we found weak expression of Oct4 and nestin and strong expression of tubulin-betaIII , P11137 , and tau . Specific markers for cholinergic , (nor)adrenergic , and GABAergic neurons or glia were weakly expressed or absent , whereas expression of factors implicated in early induction of dopaminergic neurons , TGF-beta3 and beta-catenin were present . Further analysis showed strong expression of EN-1 , c- P07949 , PTX3 , and P43354 essential for induction and survival of midbrain dopaminergic neurons , TH , P20711 , and Q05940 components of dopamine synthesis and secretion , and syntaxin1A and P60880 necessary for neurotransmitter exocytosis . This phenotype was retained throughout passages and up to the current passage 36 . Expression of neuronal and dopaminergic markers in individual AF cell lines was comparable to expression in neurons induced from ES cells and in IMR-32 and SH-SY5Y neuroblastomas . Our data show that cell lines can be derived from subcultures of amniocentesis , and are primarily composed of a population of progenitors with a phenotype similar to that of committed mesencephalic dopaminergic neurons . 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 . 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 . Central opioid inhibition of neuroendocrine stress responses in pregnancy in the rat is induced by the neurosteroid allopregnanolone . The hypothalamus-pituitary-adrenal ( Q9Y251 ) axis is the major neuroendocrine stress response system . P06850 ( P06850 ) neurons in the parvocellular paraventricular nucleus ( pPVN ) play a key role in coordinating responses of this system to stressors . The cytokine interleukin-1beta ( IL-1beta ) , mimicking infection , robustly activates these P06850 neurons via a noradrenergic input arising from the nucleus tractus solitarii ( P30990 ) . In late pregnancy , Q9Y251 axis responses to stressors , including IL-1beta , are attenuated by a central opioid mechanism that auto-inhibits noradrenaline release in the PVN . Here we show that the neuroactive progesterone metabolite allopregnanolone induces these changes in Q9Y251 responsiveness to IL-1beta in pregnancy . In late pregnancy , inhibition of 5alpha-reductase ( an allopregnanolone-synthesizing enzyme ) with finasteride restored Q9Y251 axis responses ( rapidly increased pPVN P06850 mRNA expression , DB01285 , and corticosterone secretion ) to IL-1beta . Conversely , allopregnanolone reduced Q9Y251 responses in virgin rats . In late pregnancy , activity of the allopregnanolone-synthesizing enzymes ( 5alpha-reductase and 3alpha-hydroxysteroid dehydrogenase ) was increased in the hypothalamus as was mRNA expression in the P30990 and PVN . Naloxone , an opioid antagonist , restores Q9Y251 axis responses to IL-1beta in pregnancy but had no additional effect after finasteride , indicating a causal connection between allopregnanolone and the endogenous opioid mechanism . Indeed , allopregnanolone induced opioid inhibition over Q9Y251 responses to IL-1beta in virgin rats . Furthermore , in virgin rats , allopregnanolone treatment increased , whereas in pregnant rats finasteride decreased proenkephalin-A mRNA expression in the P30990 . Thus , in pregnancy , allopregnanolone induces opioid inhibition over Q9Y251 axis responses to immune challenge . This novel opioid-mediated mechanism of allopregnanolone action may alter regulation of other brain systems in pregnancy . DB01268 treatment in pediatric patients with advanced GIST following failure of imatinib . BACKGROUND : DB01268 inhibits P10721 and other members of the split-kinase-domain family of receptor tyrosine kinases . DB01268 prolongs survival in adult patients with imatinib-resistant gastrointestinal stromal tumor ( GIST ) . We report the experience with sunitinib in pediatric patients with advanced GIST following failure of imatinib . PROCEDURE : DB01268 therapy was provided through a treatment-use protocol . Patients were 10-17 years old at enrollment . All patients had GIST resistant to imatinib therapy . DB01268 was administered daily for 4 weeks in 6-week treatment cycles . P10721 and platelet-derived growth factor receptor alpha ( P16234 ) genotyping of tumor tissue were performed . RESULTS : One patient achieved a partial response , five patients had stable disease and one patient had progressive disease on sunitinib . The duration of disease stabilization was between 7 and 21+ months , with a mean of 15 months . Time to tumor progression was longer on sunitinib than on prior imatinib treatment for five of six patients . Two patients experienced grade 3 adverse events . All other adverse events were grade 1-2 . None of the five patients tested had mutations in P10721 or P16234 . CONCLUSION : DB01268 treatment was associated with substantial initial antitumor activity and acceptable tolerability in this group of pediatric patients with imatinib-resistant GIST . 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 . Ligand-dependent platelet-derived growth factor receptor ( P09619 ) -alpha activation sensitizes rare lung cancer and sarcoma cells to P09619 kinase inhibitors . Platelet-derived growth factor ( PDGF ) receptors ( P09619 ) and their ligands play critical roles in several human malignancies . DB01268 is a clinically approved multitargeted tyrosine kinase inhibitor that inhibits vascular endothelial growth factor receptor , c- P10721 , and P09619 , and has shown clinical activity in various solid tumors . Activation of P09619 signaling has been described in gastrointestinal stromal tumors ( P16234 mutations ) as well as in chronic myeloid leukemia ( P11274 - P16234 translocation ) , and sunitinib can yield clinical benefit in both settings . However , the discovery of P09619 activating mutations or gene rearrangements in other tumor types could reveal additional patient populations who might benefit from treatment with anti- P09619 therapies , such as sunitinib . Using a high-throughput cancer cell line screening platform , we found that only 2 of 637 tested human tumor-derived cell lines show significant sensitivity to single-agent sunitinib exposure . These two cell lines [ a non-small-cell lung cancer ( NSCLC ) and a rhabdomyosarcoma ] showed expression of highly phosphorylated P16234 . In the sunitinib-sensitive adenosquamous NSCLC cell line , P16234 expression was associated with focal PFGRA gene amplification , which was similarly detected in a small fraction of squamous cell NSCLC primary tumor specimens . Moreover , in this NSCLC cell line , focal amplification of the gene encoding the P09619 ligand Q9NRA1 was also detected , and silencing P16234 or Q9NRA1 expression by RNA interference inhibited proliferation . A similar codependency on P16234 and Q9NRA1 was observed in the sunitinib-sensitive rhabdomyosarcoma cell line . These findings suggest that , in addition to gastrointestinal stromal tumors , rare tumors that show Q9NRA1 -mediated P16234 activation may also be clinically responsive to pharmacologic P16234 or Q9NRA1 inhibition . DB01268 in solid tumors . BACKGROUND : Until recently , few treatments were available for renal cell carcinoma ( RCC ) and gastrointestinal stromal tumors ( GIST ) . Several targeted agents inhibiting key pathogenetic pathways have since been developed for RCC ( sunitinib , sorafenib , bevacizumab , temsirolimus , everolimus ) and GIST ( imatinib , sunitinib ) . DB01268 is a multi-kinase inhibitor of P35968 , P09619 ( alpha,beta ) , P36888 , P10721 , P09603 and P07949 . OBJECTIVE : To summarize the literature regarding the structure , pharmacokinetics , pharmacodynamics , toxicity and current clinical use of sunitinib . Other potential roles for this drug in RCC , GIST and other tumor types will be discussed . METHODS : A literature search identified relevant (pre)clinical studies of sunitinib and other relevant agents . RESULTS/CONCLUSIONS : DB01268 revolutionized the management of advanced RCC and GIST . With the realization that cross-resistance between targeted agents is incomplete , evolving strategies include sequential treatment , concurrent treatment , and biomarker development . DB01268 also shows promise in several other tumor types that lack therapeutic options . What remains less clear is its role in tumors that are not heavily dependent on a central pathogenetic pathway , especially if effective cytotoxic therapies exist . Future clinical trials will clarify whether there is a role for sunitinib in these tumors , possibly in combination with cytotoxic agents . 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 . 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 . DB01268 synergizes the antitumor effect of cisplatin via modulation of P07992 expression in models of gastric cancer . We evaluated the effects of sunitinib monotherapy and in combination with cisplatin in human gastric cancer cell lines . DB01268 showed antiproliferative effect in gastric cancer cells line with high P16234 expression . Knockdown of P16234 showed that sunitinib sensitivity was correlated with the basal expression of P16234 . Synergistic growth inhibitory activity in combination with cisplatin was identified . We further explored how sunitinib potentiated the activity of cisplatin . We found that sunitinib treatment resulted in the down-regulation of P07992 expression via the modulation of P16234 expression in gastric cancer cells . The effect was verified via SNU484 xenograft model . Our data support the rationale of clinical trial using sunitinib in combination of cisplatin in gastric cancer . 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 . The 4q12 amplicon in malignant peripheral nerve sheath tumors : consequences on gene expression and implications for sunitinib treatment . BACKGROUND : Malignant peripheral nerve sheath tumors ( MPNST ) are highly aggressive tumors which originate from Schwann cells and develop in about 10 % of neurofibromatosis type 1 ( P21359 ) patients . The five year survival rate is poor and more effective therapies are needed . DB01268 is a drug targeting receptor tyrosine kinases ( RTK ) like PDGFRalpha , c-Kit and P35968 . These genes are structurally related and cluster on chromosomal segment 4q12 . METHODOLOGY/PRINCIPAL FINDINGS : Here we characterize this region by multiplex ligation-dependent probe amplification ( MLPA ) in MPNST . Our probe set encompasses the 3 adjacent RTK genes ( P16234 , P10721 , P35968 ) and 6 flanking genes . We found amplification of several genes within this region in a subset of MPNST and MPNST cell lines . Transcript and protein expression of P16234 matched well with its increased copy number suggesting a central role of P16234 within the amplicon . Studying the effect of sunitinib on 5 MPNST cell lines revealed that cell line S462 harboring the 4q12 amplicon was extremely sensitive to the drug with an IC50 below 1.0 microM . Moreover , sunitinib induced apoptosis and prevented PDGF-AA induced signaling via PDGFRalpha as determined by western blotting . Co-expression of P15692 and its receptor P35968 ( P35968 ) was present in MPNST cell lines suggesting an autocrine loop . We show that P15692 triggered signal transduction via the MAPK pathway , which could be blocked by sunitinib . CONCLUSIONS/SIGNIFICANCE : Since multiple receptors targeted by sunitinib are expressed or over-expressed by MPNST cells sunitinib appears as an attractive drug for treatment of MPNST patients . Presence of the 4q12 amplicon and subsequent over-expression of P16234 might serve as predictive markers for efficacy of sunitinib . 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 . [ Acute myeloid leukemia. Genetic diagnostics and molecular therapy ] . Acute myeloid leukemia ( AML ) is a genetically heterogeneous disease . The genetic diagnostics have become an essential component in the initial work-up for disease classification , prognostication and prediction . More and more promising molecular targeted therapeutics are becoming available . A prerequisite for individualized treatment strategies is a fast pretherapeutic molecular screening including the fusion genes P29590 - P10276 , Q01196 - Q06455 and Q13951 - P35749 as well as mutations in the genes P06748 , P36888 and P49715 . Promising new therapeutic approaches include the combination of all- trans retinoic acid and arsentrioxid in acute promyelocytic leukemia , the combination of intensive chemotherapy with P10721 inhibitors in core-binding factor AML and P36888 inhibitors in AML with P36888 mutation , as well as gemtuzumab ozogamicin therapy in patients with low and intermediate cytogenetic risk profiles . With the advent of the next generation sequencing technologies it is expected that new therapeutic targets will be identified . These insights will lead to a further individualization of AML therapy . 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 analysis of human glioblastomas using a genomic microarray system . Genomic microarray systems can simultaneously provide substantial genetic and chromosomal information in a relatively short time . We have analyzed genomic DNA from frozen sections of 30 cases of primary glioblastomas by GenoSensor Array 300 in order to characterize gene amplifications , gene deletions , and chromosomal information in the whole genome . Genes that were frequently amplified included P35250 / Q9UDT6 ( 63.3 % ) , P00533 ( 53.3 % ) , P05231 ( 53.3 % ) , P08183 ( P08183 ) ( 36.7 % ) , and P16234 ( 26.7 % ) . Genes that were frequently deleted included ( 56.7 % ) , P21802 ( 66.7 % ) , Q13126 ( 60.0 % ) , Q9UGM3 CDKN2A ( p16 ) / Q13126 ( 50.0 % ) , P42336 ( 43.3 % ) , and P11161 ( 43.3 % ) , but deletion of P06400 or P04637 was rarely detected . Chromosomal gains were observed frequently for 7q ( 33.3 % ) , 7p ( 20.0 % ) , and 17q ( 13.3 % ) . Loss of the 10q was frequently detected in 13 of 30 cases ( 46.7 % ) . Loss of the entire chromosome 10 was seen in 9 of 30 cases ( 30.0 % ) , and was often accompanied by P00533 amplification ( 7 cases , 77.8 % ) . The GenoSensor Array 300 proved to be useful for identification of genome-wide molecular changes in glioblastomas . The obtained microarray profile can also yield valuable insight into the molecular events underlying carcinogenesis of brain tumors and may provide clues about clinical correlations , including response to treatment .

MH_train_1120

MH_train_1120

MH_train_1120

interacts_with DB00898?

multiple_choice

Pathway based analysis of SNPs with relevance to DB00544 therapy : relation to intratumoral mRNA expression and survival . Genetic factors are thought to play a role in resistance towards chemotherapeutic agents such as 5-fluorouracil ( DB00544 ) . Approximately 30 genes are directly or indirectly involved in DB00544 metabolism , and genetic variation in any of these may contribute to anti-tumor response . Polymorphisms in these genes were analyzed in relation to tumoral mRNA levels of DB00544 metabolizing genes , response to chemotherapy and survival . A total of 21 genetic variants were studied in 35 breast cancer patients treated with 5-FluoroUracil , mitomycin ( FUMI ) and in a similar cohort of 90 doxorubicin treated breast cancer patients . Genotype distributions were compared using 109 healthy controls . No significant association was found between any polymorphisms and response to chemotherapy as measured by shrinkage of tumor . However , carriers of 3 copies of the P04818 5'UTR repeat had shorter survival than noncarriers ( p = .048 ) in the FUMI treatment group , but not in the doxorubicin treated group . Carriers of 3 copies of the repeat were also more frequently observed in both patients groups than in healthy controls ( p = .034 ) . Several highly significant associations were observed between genotypes and expression levels of DB00544 metabolizing genes . A SNP in codon 72 of P04637 was revealed to be a key regulator of DB00544 metabolizing genes such as P00374 and P42898 , constituting 50 % of all significant associations observed after FUMI therapy . These data suggest that 3 copies of the P04818 5'UTR repeat may give a treatment specific reduced survival in breast cancer patients , and that P04637 may have a direct , allele specific , role in DB00544 mediated response . 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 . Low ethanol concentration alters P30532 RNA levels during early human development . DB00898 use is common and consumption during pregnancy has been shown to lead to a myriad of physical and neurologic abnormalities commonly referred to as fetal alcohol spectrum disorder . Substance addiction , which includes alcohol , has been shown to involve the major nicotinic acetylcholine receptor subunit P30532 . Using human embryonic stem cells as a model of early human development , we show that low concentrations of ethanol ( 20mM ) can alter the expression of P30532 . Changes in P30532 expression is linked to altered GABA and DB01221 receptor expression , as well as abnormal development of the frontal cortex . These results suggest that alcohol exposure can alter early neurologic development , which may favor addiction and other developmental abnormalities in unborn children . 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 . 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 . Chromosome 8p as a potential hub for developmental neuropsychiatric disorders : implications for schizophrenia , autism and cancer . Defects in genetic and developmental processes are thought to contribute susceptibility to autism and schizophrenia . Presumably , owing to etiological complexity identifying susceptibility genes and abnormalities in the development has been difficult . However , the importance of genes within chromosomal 8p region for neuropsychiatric disorders and cancer is well established . There are 484 annotated genes located on 8p ; many are most likely oncogenes and tumor-suppressor genes . Molecular genetics and developmental studies have identified 21 genes in this region ( ADRA1A , O15013 , Q15822 , Q15825 , Q05901 , Q9UBT3 , Q16555 , Q06889 , O60258 , Q9NP95 , P11362 , FZD3 , LDL , NAT2 , P07197 , Q02297 , PCM1 , P00750 , P48454 , Q8N474 and P54219 / P54219 ) that are most likely to contribute to neuropsychiatric disorders ( schizophrenia , autism , bipolar disorder and depression ) , neurodegenerative disorders ( Parkinson 's and Alzheimer 's disease ) and cancer . Furthermore , at least seven nonprotein-coding RNAs ( microRNAs ) are located at 8p . Structural variants on 8p , such as copy number variants , microdeletions or microduplications , might also contribute to autism , schizophrenia and other human diseases including cancer . In this review , we consider the current state of evidence from cytogenetic , linkage , association , gene expression and endophenotyping studies for the role of these 8p genes in neuropsychiatric disease . We also describe how a mutation in an 8p gene ( Fgf17 ) results in a mouse with deficits in specific components of social behavior and a reduction in its dorsomedial prefrontal cortex . We finish by discussing the biological connections of 8p with respect to neuropsychiatric disorders and cancer , despite the shortcomings of this evidence . Genetic susceptibility to aseptic loosening following total hip arthroplasty : a systematic review . Introduction Aseptic loosening is the most common cause of total hip arthroplasty ( DB00382 ) failure and revision surgery . Genetic polymorphisms could be determinant factors for implant loosening . Source of data We performed a comprehensive search of Medline , CINAHL , Googlescholar , Embase and Cochrane databases , using various combinations of the keyword terms ' aseptic loosening ' , ' gene ' , ' hip arthoplasty ' , ' genetics ' , ' loosening ' . Twelve studies detailing the genetic investigation of patients with aseptic loosening of a DB00382 were identified . Areas of agreement SNPs of GNAS1 , P01375 -238 A allele , P01375 -α promoter ( -308G→A ) transition , P05231 -174 G allele , interleukin ( IL ) -6 ( -597 ) and ( -572 ) , P03956 -promoting gene , C/C genotype for the P03956 , P50281 , P08253 , transforming growth factor-beta1 signal sequence ( 29T→C ) transitions , A/A genotype for the O00300 -163 , and P11226 were overexpressed in patients with aseptic loosening and periprosthetic osteolysis . Areas of controversy Data from single centre studies do not allow one to compare the results of different studies . Conclusion Several gene pathways and genes contribute to the genetic susceptibility to aseptic loosening following DB00382 . Further studies will enhance the understanding of prosthesis failure , and may inform and direct pharmaceutical interventions . Growing points Further multi-centre prospective studies are necessary to confirm the general validity of the findings reported . Single-centre findings should be replicated in other centres and populations to open new avenues for pre-surgical genetic testing and to investigate immune response modulation in DB00382 . Areas timely for developing research Research in this field could lead to better understanding of mechanisms behind aseptic osteolysis , and improve the results of DB00382 . 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 . Proteomics screen to reveal molecular changes mediated by C722G missense mutation in P08172 gene . Previously , we reported a missense mutation ( C722G ) in the M2-muscarinic acetylcholine receptor ( P08172 ) gene associated with familial dilated cardiomyopathy . However , the exact molecular mechanisms by the related protein changes of P08172 -C722G mutation induced are still unclear . P08172 and P08172 -C722G lentiviral vector was infected to CHO cells . Proteomic analysis by label-free shotgun strategy and the STRING 9.0 software were performed . A total of 102 proteins with at least 2-fold change in the P08172 -C722G group were identified , 42 proteins were up-regulated , whereas 57 were down-regulated . These altered proteins belong to three broad functional categories : ( i ) metabolic ( e.g. O00154 , Malate dehydrogenase ) ; ( ii ) cytoskeletal ( e.g. Actin-related protein , P60660 and P12814 ) and ( iii ) stress response ( e.g. heat shock protein 70 , P61026 ) . Interestingly , the marked differences in the expression of selected eight proteins ( change > 4.0-fold ) , were connected with many proteins related to apoptosis and immune/inflammatory response such as : P01100 , Q07812 , MYC , P04637 and P05231 . This novel study demonstrated for the first time a full-scale screening of the proteomics research by P08172 -C722G mutation and profiled 102 changed proteins , of which , eight might be critical in cardiac dysfunction for future mapping . SIGNIFICANCE : It was a full-scale screening of the proteomics research by P08172 -C722G mutation . These proteins might serve as valuable biomarkers that could predict the presence of a precursor field . These proteins might serve to further explore the pathophysiological mechanisms in familial DCM patients with C176W mutation . 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 . 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 . 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 . The heteromeric GABA-B receptor recognizes G-protein alpha subunit C-termini . The recently cloned GABA-B receptors are related to the metabotropic glutamate receptors ( mGlu receptors ) , the Ca2+-sensing receptor and one group of vomeronasal receptors . The GABA-B receptors likely function in a heterodimeric form , constituted of Q9UBS5 and O75899 . This novel feature in the G-protein coupled receptors ( GPCRs ) structure raises questions as to the mechanism of recognition of G-proteins by such receptors . In the present study we show that the Q9UBS5 and BR2 subunits form a functional receptor that recognizes the extreme C-termini of the G alpha i and G alpha o proteins when expressed in HEK293 cells . Indeed , heteromeric Q9UBS5 /BR2 receptors do not activate P98160 when co-expressed with G alpha q , but do so when co-expressed with the chimeric G alpha qi5 or G alpha qo5 subunits , the G alpha q subunit in which the 5 C-terminal residues are those of G alpha i or G alpha o , respectively . Interestingly , the heteromeric GABA-B receptor did not activate the chimeric G alpha qz5 subunit that contains the 5 C-terminal residues of G alpha z . Among the three residues that are distinct between G alpha qo5 and G alpha qz5 ( at position -5 , -4 and -1 ) , the amino acid residue at position -4 of G alpha o proteins is critical for specifying the coupling selectivity with the receptor and residue -5 influences the coupling efficacy . Interestingly , these findings correspond to data obtained with the Q14416 receptor , a distant relative of GABA-B proteins . This shows that the same molecular determinants of the G-protein alpha-subunits are involved in the specific recognition of both the heteromeric GABA-B receptors and the other GPCRs . Support for association between ADHD and two candidate genes : NET1 and P21728 . Attention deficit hyperactivity disorder ( ADHD ) is a common , multifactorial disorder with significant genetic contribution . Multiple candidate genes have been studied in ADHD , including the norepinephrine transporter ( NET1 ) and dopamine D1 receptor ( P21728 ) . NET1 is implicated in ADHD because of the efficacy of atomoxetine , a selective noradrenergic reuptake inhibitor , in the treatment of ADHD . P21728 is primarily implicated through mouse models of ADHD . DNA from 163 ADHD probands , 192 parents , and 129 healthy controls was used to investigate possible associations between ADHD and polymorphisms in 12 previously studied candidate genes ( P28222 , 5- Q13049 , P28335 , P08913 , P43681 , P21964 , Q01959 , P21728 , P21917 , P21918 , NET1 , and P60880 ) . Analyses included case-control and family-based methods , and dimensional measures of behavior , cognition , and anatomic brain magnetic resonance imaging ( Q9BWK5 ) . Of the 12 genes examined , two showed a significant association with ADHD . Transmission disequilibrium test ( P04053 ) analysis revealed significant association of two NET1 single nucleotide polymorphisms ( SNPs ) with ADHD ( P < or = 0.009 ) ; case-control analysis revealed significant association of two P21728 SNPs with ADHD ( P < or = 0.008 ) . No behavioral , cognitive , or brain Q9BWK5 volume measurement significantly differed across NET1 or P21728 genotypes at an alpha of 0.01 . This study provides support for an association between ADHD and polymorphisms in both NET1 and P21728 ; polymorphisms in ten other candidate genes were not associated with ADHD . Because family-based and case-control methods gave divergent results , both should be used in genetic studies of ADHD . 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 . Differential toxicity biomarkers for irinotecan- and oxaliplatin-containing chemotherapy in colorectal cancer . PURPOSE : DB00526 or irinotecan is usually administered jointly with fluoropyrimidines in colorectal cancer patients treated with chemotherapy . Both drugs have different toxicity patterns . Biomarkers for predicting high-risk severe adverse reactions can help select the best treatment . METHODS : A retrospective analysis of 106 colorectal cancer patients receiving an oxaliplatin-based treatment and 56 receiving an irinotecan-based treatment was performed . One copy number variant ( P30711 ) and nine polymorphisms in irinotecan and oxaliplatin metabolism , transport or DNA repair genes ( P08183 , P22309 , P18887 , P07992 , P18074 , P09211 ) were genotyped by SNaPshot , polymerase chain reactions ' length fragments , or copy number assays . RESULTS : In irinotecan-treated patients , T allele of ABCB1C1236T SNP was associated with a lower risk of asthenia ( OR = 0.047 ; 95 % CI = 0.004–0.493 ; P = 0.011 ) and Tallele of P08183 C3435T SNP was associated with a lower risk of diarrhea ( OR = 0.177 ; 95 % CI = 0.034–0.919;P = 0.039 ) , and individuals with two copies of P30711 gene had a lower risk for asthenia ( OR = 0.093 ; 95 % CI = 0.011–0.794 ; P = 0.030 ) . In oxaliplatin-treated patients , carriers of one or two T variants of Asn118Asn P07992 SNP had a lower risk for neutropenia ( OR = 0.205 ; 95 % CI = 0.061–0.690 ; P = 0.01 ) [ corrected ] . CONCLUSIONS : These biomarkers could help oncologists select the best treatment by reducing toxicity associated with irinotecan or oxaliplatin in colorectal cancer patients , thus increasing their quality of life . Association study of 45 candidate genes in nicotine dependence in Han Chinese . Numerous genetic linkages , association studies have been performed in different ethnic groups and revealed many susceptibility loci and genes for nicotine dependence . However , limited similar researches were performed in Han Chinese . This study was designed to investigate the association of candidate genes with nicotine dependence in Han Chinese . We genotyped 384 SNPs within 45 candidate genes with nicotine dependence in a Han Chinese population consisting 223 high nicotine dependent subjects and 257 low nicotine dependent subjects by employing GoldenGate genotyping assay ( Illumina ) . Following association analysis was performed using PLINK software . Individual SNP-based association analysis revealed that nine SNPs located in P35462 ( rs2630351 ) , P21918 ( rs1967550 ) , Q9Y6R4 ( rs2314378 ) , DDC ( rs11575461 ) , Q05901 ( rs4954 ) , O75899 ( rs2779562 ) , P14416 ( rs11214613 and rs6589377 ) and P43681 ( rs2236196 ) were significantly associated with FTND after correction for multiple testing with the p values from 2.59×10(-7) to 9.99×10(-5) . Haplotype-based association analysis revealed haplotype G-A-A formed by rs2630351 , rs167771 and rs324032 and haplotype G-G-G-A formed by rs3773678 , rs2630349 , rs2630351 and rs167771 in P35462 ; haplotype of G-A formed by rs2779562 and rs2808566 in O75899 and haplotype of T-T-A-G-A formed by rs6832644 , rs4057797 , rs9764 , rs4552421 and rs10033119 in P25929 are associated with FTND ( p=3.61×10(-7)-8.78×10(-6) ) . Our results provided confirmation of the previous findings that P14416 , P35462 , DDC , Q05901 , O75899 and P43681 are associated with nicotine dependence . Furthermore , we for the first time report a significant association between nicotine dependence and P21918 , Q9Y6R4 and P25929 . These findings need independent replication in the future studies . Determinants in the β and δ subunit cytoplasmic loop regulate Golgi trafficking and surface expression of the muscle acetylcholine receptor . The molecular determinants that govern nicotinic acetylcholine receptor ( AChR ) assembly and trafficking are poorly defined , and those identified operate largely during initial receptor biogenesis in the endoplasmic reticulum . To identify determinants that regulate later trafficking steps , we performed an unbiased screen using chimeric proteins consisting of P01730 fused to the muscle AChR subunit cytoplasmic loops . In P06681 mouse muscle cells , we found that P01730 -β and δ subunit loops were expressed at very low levels on the cell surface , whereas the other subunit loops were robustly expressed on the plasma membrane . The low surface expression of P01730 -β and δ loops was due to their pronounced retention in the Golgi apparatus and also to their rapid internalization from the plasma membrane . Both retention and recovery were mediated by the proximal 25-28 amino acids in each loop and were dependent on an ordered sequence of charged and hydrophobic residues . Indeed , βK353L and δK351L mutations increased surface trafficking of the P01730 -subunit loops by > 6-fold and also decreased their internalization from the plasma membrane . Similarly , combined βK353L and δK351L mutations increased the surface levels of assembled AChR expressed in P29320 cells to 138 % of wild-type levels . This was due to increased trafficking to the plasma membrane and not decreased AChR turnover . These findings identify novel Golgi retention signals in the β and δ subunit loops that regulate surface trafficking of assembled AChR and may help prevent surface expression of unassembled subunits . Together , these results define molecular determinants that govern a Golgi-based regulatory step in nicotinic AChR trafficking . DB00559 , an endothelin receptor antagonist , ameliorates collagen-induced arthritis : the role of P01375 -α in the induction of endothelin system genes . OBJECTIVE : Endothelins ( ETs ) are involved in several inflammatory events . The present study investigated the efficacy of DB00559 , a dual P25101 /ETB receptor antagonist , in collagen-induced arthritis ( CIA ) in mice . TREATMENT : CIA was induced in DBA/1J mice . Arthritic mice were treated with DB00559 ( 100 mg/kg ) once a day , starting from the day when arthritis was clinically detectable . METHODS : CIA progression was assessed by measurements of visual clinical score , paw swelling and hypernociception . Histological changes , neutrophil infiltration and pro-inflammatory cytokines were evaluated in the joints . Gene expression in the lymph nodes of arthritic mice was evaluated by microarray technology . PreproET-1 mRNA expression in the lymph nodes of mice and in peripheral blood mononuclear cells ( PBMCs ) was evaluated by real-time PCR . The differences were evaluated by one-way Q9UNW9 or Student 's t test . RESULTS : Oral treatment with DB00559 markedly ameliorated the clinical aspects of CIA ( visual clinical score , paw swelling and hyperalgesia ) . DB00559 treatment also reduced joint damage , leukocyte infiltration and pro-inflammatory cytokine levels ( IL-1β , TNFα and Q16552 ) in the joint tissues . Changes in gene expression in the lymph nodes of arthritic mice returned to the levels of the control mice after DB00559 treatment . PreproET mRNA expression increased in PBMCs from rheumatoid arthritis ( RA ) patients but returned to basal level in PBMCs from patients under anti- P01375 therapy . In-vitro treatment of PBMCs with TNFα upregulated ET system genes . CONCLUSION : These findings indicate that ET receptor antagonists , such as DB00559 , might be useful in controlling RA . Moreover , it seems that ET mediation of arthritis is triggered by TNFα . GABA Receptors Genes Polymorphisms and DB00898 Dependence : No Evidence of an Association in an Italian Male Population . OBJECTIVE : The genes encoding for gamma-aminobutyric acid ( GABA ) A and B receptors may be considered as candidates for alcoholism ; genetic alterations at this level may produce structural and functional diversity and thus play a role in the response to alcohol addiction treatment . To investigate these aspects further , we conducted a preliminary genetic association study on a population of Italian male alcohol addicts , focusing on GABA A and B receptors . METHODS : A total of 186 alcohol-dependent subjects ( in the first phase 139 , then 47 more samples ) and 182 controls were genotyped for 25 single nucleotide polymorphisms ( SNPs ) of genes encoding the alpha-1 subunit of GABA A receptor ( P14867 ) and subunits 1 and 2 of GABA B receptor ( Q9UBS5 and O75899 ) . The chi-squared test for allele and genotype distributions and Hardy-Weinberg equilibrium analysis of both subjects and controls were performed . Bonferroni 's correction for multiple comparisons was applied . RESULTS : Preliminary results comparing 139 alcohol-dependent subjects and 182 controls showed differences in genotype distribution in the former for SNP rs29253 , located in the intron region of the Q9UBS5 gene . In order to clarify the meaning of this association , 47 more samples from alcohol-dependent subjects were tested for this SNP only : the previously found association was not confirmed . CONCLUSION : The lack of significant differences between the two groups does not provide evidence that GABRA 1 and Q9UBS5 and 2 genes are candidates for alcoholism in this population . Further studies with larger samples are needed , together with investigation of other components of the GABA pathway . 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 . Differences in immune cell function between tuberculosis positive and negative Asian elephants . Tuberculosis is an important health concern for Asian elephant ( Elephas maximus ) populations worldwide , however , mechanisms underlying susceptibility to Mycobacterium tuberculosis are unknown . Proliferative responses assessed via brominated uridine incorporation and cytokine expression measured by real-time RT-PCR were evaluated in peripheral blood mononuclear cell ( PBMC ) cultures from 8 tuberculosis negative and 8 positive Asian elephants . Cultures were stimulated with Mycobacterium bovis purified protein derivative ( PPD-B ) , M. tuberculosis culture filtrate protein ( P27918 ) -10 , and Mycobacterium avium PPD ( PPD-A ) . Following stimulation with PPD-B , proliferation was higher ( α = 0.005 ) in positive samples ; no significant differences were detected following P27918 -10 or PPD-A stimulation . P01375 ( P01375 ) -α , interleukin ( IL ) -12 , and interferon ( IFN ) -γ expression was greater in samples from positive elephants following stimulation with PPD-B ( α = 0.025 ) and P27918 -10 ( α = 0.025 P01375 -α and IL-12 ; α = 0.005 IFN-γ ) . Stimulation with PPD-A also produced enhanced IL-12 expression in positive samples ( α = 0.025 ) . Findings suggested that differences in immune cell function exist between tuberculosis positive and negative elephants . Proliferative responses and expression of P01375 -α , IL-12 , and IFN-γ in response to stimulation with PPD-B and P27918 -10 differ between tuberculosis positive and negative elephants , suggesting these parameters may be important to tuberculosis immunopathogenesis in this species . Nicotinic acetylcholine receptors containing the α6 subunit contribute to ethanol activation of ventral tegmental area dopaminergic neurons . DB00184 and alcohol are often co-abused suggesting a common mechanism of action may underlie their reinforcing properties . Both drugs acutely increase activity of ventral tegmental area ( VTA ) dopaminergic ( DAergic ) neurons , a phenomenon associated with reward behavior . Recent evidence indicates that nicotinic acetylcholine receptors ( nAChRs ) , ligand-gated cation channels activated by ACh and nicotine , may contribute to ethanol-mediated activation of VTA DAergic neurons although the nAChR subtype(s) involved has not been fully elucidated . Here we show that expression and activation of nAChRs containing the α6 subunit contribute to ethanol-induced activation of VTA DAergic neurons . In wild-type ( WT ) mouse midbrain sections that contain the VTA , ethanol ( 50 or 100 mM ) significantly increased firing frequency of DAergic neurons . In contrast , ethanol did not significantly increase activity of VTA DAergic neurons in mice that do not express Q15825 , the gene encoding the α6 nAChR subunit ( α6 knock-out ( KO ) mice ) . DB00898 -induced activity in WT slices was also reduced by pre-application of the α6 subtype-selective nAChR antagonist , α-conotoxin MII[E11A] . When co-applied , ethanol potentiated the response to ACh in WT DAergic neurons ; whereas co-application of ACh and ethanol failed to significantly increase activity of DAergic neurons in α6 KO slices . Finally , pre-application of α-conotoxin MII[E11A] in WT slices reduced ethanol potentiation of ACh responses . Together our data indicate that α6-subunit containing nAChRs may contribute to ethanol activation of VTA DAergic neurons . These receptors are predominantly expressed in DAergic neurons and known to be critical for nicotine reinforcement , providing a potential common therapeutic molecular target to reduce nicotine and alcohol co-abuse . 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 . Strong bias in the location of functional promoter polymorphisms . A considerable proportion of heritable human phenotypic variation is thought to result from altered gene expression . Unfortunately , it is currently impossible to use bioinformatic analysis to discriminate between DNA sequence variants that are likely to influence gene expression and those that are not . In an attempt to define some of the characteristics of promoter polymorphisms with functional effects on gene expression , we examined 674 haplotypes representing 247 unique gene promoters using a standardized reporter gene assay system . Sequence variants that altered gene expression by 1.5-fold or more were strongly biased toward a location in the core and proximal promoter regions , 50 % being within the first 100 bases 5' to the transcription start site . No bias was seen in the allele frequencies of functional and nonfunctional sequence variants . Only 33 % of the functional variants were found in known consensus transcription factor binding sequences or motifs , which suggests that either there are many unknown transcription factor binding motifs or other , unknown mechanisms are involved . The genes with functional polymorphisms that are reported here for the first time include AGTRL2 , CAT , P30532 , P08311 , P10635 , DLD , P07992 , P14867 , O00591 , P31942 , O00291 , IGKV1-9 , Q99712 , Q9Y257 , P06870 , P08118 , Q99972 , P49146 , Q99466 , P19652 , P36955 , Q14761 , Q13007 ( Q13007 ) , P50225 , and P16473 . 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 . 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 . Aberrant DNA methylation distinguishes hepatocellular carcinoma associated with HBV and HCV infection and alcohol intake . BACKGROUND & AIMS : Hepatocellular carcinoma ( HCC ) is one of the most frequent human cancers and a major cause of cancer-related death worldwide . The major risk factors for developing HCC are infection by hepatitis B virus ( HBV ) and hepatitis C virus ( HCV ) , chronic alcoholism , and aflatoxins ; however , critical gene targets remain largely unknown . Herein , we sought to establish DNA methylation patterns in HCC and corresponding cirrhotic tissues and to identify DNA methylation changes associated with major risk factors . METHODS : We have established assays for quantitative analysis of DNA methylation levels in a panel of seven cancer-associated genes and repetitive elements , and combined these assays with a series of HCC tumors , associated with major risk factors , collected from two different geographical areas . RESULTS : We found a high frequency of aberrant hypermethylation of specific genes ( RASSF1A , P09211 , P32297 , and DOK1 ) in HCC tumors as compared to control cirrhotic or normal liver tissues , suggesting that aberrant hypermethylation exhibits non-random and tumor-specific patterns in HCC . Importantly , our analysis revealed an association between alcohol intake and the hypomethylation of P16455 and between hypermethylation of P09211 and HBV infection , indicating that hypermethylation of the genes analyzed in HCC tumors exhibits remarkably distinct patterns depending on associated risk factors . CONCLUSIONS : This study identifies aberrant DNA methylation of specific cellular genes in HCC and the major risk factors associated with these changes , providing information that could be exploited for biomarker discovery in clinics and molecular epidemiology . 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 . Innovative approaches for the development of antidepressant drugs : current and future strategies . Depression is a highly debilitating disorder that has been estimated to affect up to 21 % of the world population . Despite the advances in the treatment of depression with selective serotonin reuptake inhibitors ( SSRIs ) and serotonin and norepinephrine reuptake inhibitors ( SNRIs ) , there continue to be many unmet clinical needs with respect to both efficacy and side effects . These needs range from efficacy in treatment resistant patients , to improved onset , to reductions in side effects such as emesis or sexual dysfunction . To address these needs , there are numerous combination therapies and novel targets that have been identified that may demonstrate improvements in one or more areas . There is tremendous diversity in the types of targets and approaches being taken . At one end of a spectrum is combination therapies that maintain the benefits associated with SSRIs but attempt to either improve efficacy or reduce side effects by adding additional mechanisms ( P08908 , P28222 , P28221 , P28335 , alpha-2A ) . At the other end of the spectrum are more novel targets , such as neurotrophins ( P23560 , IGF ) , based on recent findings that antidepressants induce neurogenesis . In between , there are many approaches that range from directly targeting serotonin receptors ( P28335 , P50406 ) to targeting the multiplicity of potential mechanisms associated with excitatory ( glutamate , DB01221 , Q14416 , P41594 ) or inhibitory amino acid systems ( GABA ) or peptidergic systems ( neurokinin 1 , DB05394 1 , melanin-concentrating hormone 1 , V1b ) . The present review addresses the most exciting approaches and reviews the localization , neurochemical and behavioral data that provide the supporting rationale for each of these targets or target combinations . 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 . 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 . 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 . 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 . 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 . 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 . In vivo selection for human and murine hematopoietic cells transduced with a therapeutic P16455 lentiviral vector that inhibits HIV replication . We have developed an HIV-based lentiviral vector , DB05251 , which efficiently transduces human P28906 + progenitors and P01730 + T lymphocytes . DB05251 contains an antisense sequence against the HIV envelope and is currently being evaluated for safety in a clinical trial for treatment of HIV . Selective outgrowth of transduced hematopoietic cells in vivo is anticipated to increase the therapeutic efficacy of this treatment by maximizing the persistence of virus-resistant cells in the body . Although HIV resistance is selective , additional selection may aid in treatment efficacy due to the vast quantity of target cells . Therefore , we engineered DB05251 to express the P140K P16455 gene to drive potent drug-mediated in vivo selection for transduced hematopoietic long-term repopulating cells . Suboptimally transduced T cell cultures treated with O6-benzylguanine and DB00262 were selected from 3 to 100 % , and after selection cultures did not support HIV replication . Primary P28906 + progenitors derived from G- P04141 -mobilized peripheral blood were transduced at 27 to 35 % efficiency . Approximate sixfold selection was observed for transduced P28906 + progenitors , colony-forming units , and long-term culture-initiating cells . Multilineage in vivo selection was demonstrated for transduced murine hematopoietic cells in human P28906 (+)-derived hematopoietic cells in NOD-SCID mice . These results establish efficient ex vivo and in vivo selection for hematopoietic cells transduced with lentiviral vectors and support the potential therapeutic benefit of this strategy in human gene therapy . P01375 alpha mediates GABA(A) receptor trafficking to the plasma membrane of spinal cord neurons in vivo . The proinflammatory cytokine TNFα contributes to cell death in central nervous system ( CNS ) disorders by altering synaptic neurotransmission . TNFα contributes to excitotoxicity by increasing P42262 -lacking AMPA receptor ( AMPAR ) trafficking to the neuronal plasma membrane . In vitro , increased AMPAR on the neuronal surface after TNFα exposure is associated with a rapid internalization of GABA(A) receptors ( GABA(A)Rs ) , suggesting complex timing and dose dependency of the CNS 's response to TNFα . However , the effect of TNFα on GABA(A)R trafficking in vivo remains unclear . We assessed the effect of TNFα nanoinjection on rapid GABA(A)R changes in rats ( N = 30 ) using subcellular fractionation , quantitative western blotting , and confocal microscopy . GABA(A)R protein levels in membrane fractions of TNFα and vehicle-treated subjects were not significantly different by Western Blot , yet high-resolution quantitative confocal imaging revealed that TNFα induces GABA(A)R trafficking to synapses in a dose-dependent manner by 60 min . TNFα-mediated GABA(A)R trafficking represents a novel target for CNS excitotoxicity . 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 . 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 . 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 . 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 . Neuronal localization of glutamate receptor subunits in the basolateral amygdala . Antibodies to the Q05586 glutamate receptor subunit and the GluR1 and GluR2/3 subunits of the AMPA glutamate receptor were used to localize these receptor components in the basolateral amygdala ( P00519 ) of the rat and monkey . A similar localization pattern was observed in both species . Pyramidal neurons exhibited high levels of Q05586 and GluR2/3 immunoreactivity ( ir ) , but low levels of GluR1-ir . Some non-pyramidal cells exhibited high levels of Q05586 -ir or GluR1-ir , but none exhibited significant levels of GluR2/3-ir . This differential localization of receptor subunits suggests that glutamate receptors will exhibit specific functional properties in distinct subpopulations of P00519 neurons . Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking . Binge drinking of alcoholic beverages among adolescents is a common practice that can have serious health consequences . DB00898 is a potent immunomodulator that alters a wide range of immune responses . However , it is unclear whether there is a differential immune response to alcoholic beverages with a high versus low concentration of ethanol . In this study , we used a PCR array containing 46 primer pairs of selected genes to compare mRNA expression in the spleen , an immune system organ , of adolescent rats following binge drinking of alcohol solutions containing either 20 % or 52 % ethanol ( v/v , 4.8 g/kg daily dosage ) , or water ( control ) for 3 d . We found that , expression of IL-1β , P05231 , P13500 , and GABA(A) receptor α2 subunit in the spleen were decreased , and P41594 and P46098 receptor expression were increased after administration of an ethanol solution containing 52 % ethanol , but not one with 20 % ethanol . Our data suggest that alcohol-mediated immunomodulatory effects are , in part , dependent on the ethanol by volume concentration . This is the first study to show that exposure to a high ethanol percentage beverage can have more profound effects on immune responses than one with a low percentage of ethanol . 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 . [ 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 . 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 . 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 . The P08908 -receptor agonist flibanserin reduces drug-induced dyskinesia in O75916 -deficient mice . Drug-induced dyskinesia is a major complication of dopamine replacement therapy in advanced Parkinson 's disease consisting of dystonia , chorea and athetosis . Agonists at P08908 -receptors attenuate levodopa-induced motor complications in non-human primates . Mice with increased dopamine D2 receptor ( P14416 ) signalling due to the lack of expression of the regulator of G-protein signalling 9 ( O75916 ) also develop dyskinesia following levodopa treatment . We investigated whether the P08908 -receptor agonist flibanserin compared with buspirone reduces motor abnormalities induced by levodopa or quinelorane , a selective dopamine D2-receptor agonist . Following dopamine depletion via reserpine , 40 mice ( 20 wild-type and 20 O75916 knock-out ) were treated with flibanserin or buspirone in combination with levodopa or quinelorane . Motor behaviour was analysed using open field analysis . O75916 knock-out mice displayed significantly more drug-induced dystonia ( p < 0.04 ; t test ) than wild type . In quinelorane-treated wild-type mice flibanserin as well as buspirone significantly reduced dystonia ( p < 0.05 ) . In O75916 knock-out animals again both reduced quinelorane-induced dystonia . However , flibanserin was significantly more effective ( p = 0.003 ) . Following reserpine pretreatment and administration of levodopa wild-type and Q99697 9 knock-out mice showed mild to moderate dystonia . Surprisingly , 10 mg/kg buspirone increased dystonia in both animal groups , whereas it was decreased by 10 mg/kg flibanserin . However , compared with levodopa alone only the increase of dystonia by buspirone was significant ( p < 0.04 ) . DB04908 showed promising antidyskinetic effects in a model of drug-induced dyskinesia . Our data underline the possible benefit of P08908 agonists in drug-induced dyskinesia . 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 . 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 . Emergence of FGFR family gene fusions as therapeutic targets in a wide spectrum of solid tumours . The emergence of fibroblast growth factor receptor ( FGFR ) family fusions across diverse cancers has brought attention to FGFR-derived cancer therapies . The discovery of the first recurrent FGFR fusion in glioblastoma was followed by discoveries of FGFR fusions in bladder , lung , breast , thyroid , oral , and prostate cancers . Drug targeting of FGFR fusions has shown promising results and should soon be translating into clinical trials . FGFR fusions form as a result of various mechanisms – predominantly deletion for P11362 , translocation for P21802 , and tandem duplication for P22607 . The ability to exploit the unique targetability of FGFR fusions proves that FGFR-derived therapies could have a promising future in cancer therapeutics . Drug targeting of fusion genes has proven to be an extremely effective therapeutic approach for cancers such as the recurrent BCR– P00519 fusion in chronic myeloid leukaemia . The recent discovery of recurrent FGFR family fusions in several cancer types has brought to attention the unique therapeutic potential for FGFR-positive patients . Understanding the diverse mechanisms of FGFR fusion formation and their oncogenic potential will shed light on the impact of FGFR-derived therapy in the future . 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 . 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 . DNA base excision repair gene polymorphisms modulate human cognitive performance and decline during normal life span . To test the hypothesis that single nucleotide polymorphisms ( SNPs ) in DNA repair genes are associated with cognitive performance during normal aging , the relationship between SNPs in selected exons in DNA base excision repair ( BER ) genes and cognitive performance was examined in 712 healthy Norwegian individuals aged 20-75 years . SNPs examined included PolB(Pro242Arg) , hOGG1(Ser326Cys) , MutYH ( Met22Val ) , MutYH(His324Gln) , P27695 (Gln51His) , P27695 (Glu148Asp) , P18887 (Lys298Asn) , P18887 (Arg7Leu) , Q96FI4 (Asp252Asn) , and Q969S2 (Arg257Leu) . P18887 (Arg7Leu) and PolB(Pro242Arg) were characterized by single nucleotide variations ( ≤0.1 % homozygote SNPs ) . hOGG1(Ser326Cys) ( DB00133 / DB00151 40.8 % / DB00151 / DB00151 5.7 % ) , MutYH(His324Gln) ( DB00117 /Gln37 % /Gln/Gln 6.0 % ) and P27695 (Glu148Asp) ( DB00142 / DB00128 51.3 % / DB00128 / DB00128 23.0 % ) were characterized by higher SNP frequencies . MutYH(Met22Val) , P27695 (Gln51His) and Q969S2 (Arg257Leu) occurred at intermediate SNP frequencies of 11.5 , 7.6 and 5.3 % , respectively . Interestingly , hOGG1(Ser326Cys) and P27695 (Gln51His) had genotype by age interactions with general cognitive function , reasoning , control and speed of processing in cross-sectional analysis and a significant effect on longitudinal decline . Dispersed association effects involving MutYH(His324Gln) , MutYH(Met22Val) , PolB(Pro242Arg) and Q969S2 (Arg257Leu) were also detected when P02649 or P43681 , were included in the statistical model , a result consistent with proposed involvement of the latter markers in human cognitive decline and/or function . In summary , the results support the notion that polymorphisms in BER genes modulate cognitive performance in healthy elderly individuals .

MH_train_1121

MH_train_1121

MH_train_1121

interacts_with DB06695?

multiple_choice

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 . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . Mechanisms of ibrutinib resistance in chronic lymphocytic leukaemia and non-Hodgkin lymphoma . Q06187 ( Q06187 ) , a mediator of B-cell receptor ( P11274 ) signalling , has been implicated in the pathogenesis of chronic lymphocytic leukaemia ( CLL ) and other B-cell malignancies . DB09053 is an orally bioavailable and highly specific Q06187 inhibitor that was recently approved for treatment of patients with recurrent CLL and mantle cell lymphoma ( Q8WXI8 ) . In addition , ibrutinib has shown efficacy in subsets of patients with diffuse large B cell lymphoma ( DLBCL ) and Waldenstrom macroglobulinaemia ( WM ) . However , despite ibrutinib 's activity in multiple B-cell malignancies , cases of primary and secondary resistance have emerged . The overall reported frequency of resistance is low , but follow-up in many trials was short , and we predict that the incidence of observed resistance will increase as clinical use outside clinical trials expands over time . Mutations within Q06187 have been described and clearly interfere with drug binding ; however , there are also emerging alternative mechanisms that bypass Q06187 entirely and offer new opportunities for other targeted agents . Improved understanding of mechanisms of primary and secondary resistance is essential to developing appropriate therapeutic strategies to both prevent and address resistance . This review provides a comprehensive analysis of ibrutinib resistance in CLL , Q8WXI8 , DLBCL and WM and considers potential strategies for further study . 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 . 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 . Long-term stability of international reference preparations for thromboplastins . Three certified reference materials for thromboplastins are available from the Community Bureau of Reference ( P11274 ) of the European Commission for calibration of commercial thromboplastins used for control of oral anticoagulant therapy . The long-term stability of these reference materials has been monitored by two independent laboratories , using deep-frozen and lyophilized plasma samples . P00734 times and prothrombin time ratios measured on 19 occasions in the period 1981-86 have been analysed for trend with time . Although significant trends of prothrombin time and ratio ( P less than 0.05 ) were observed , a consistent pattern of trends could not be recognized . The significant trends of prothrombin time and prothrombin time ratio are most probably due to changes in local laboratory conditions . There is no indication that the reference materials have deteriorated since the beginning of the study . It is recommended that long-term stability monitoring of thromboplastins be performed by at least two laboratories simultaneously .

MH_train_1122

MH_train_1122

MH_train_1122

interacts_with DB00641?

multiple_choice

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 . 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 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 . Anti-atherogenic and anti-angiogenic activities of polyphenols from propolis . Propolis is a polyphenol-rich resinous substance extensively used to improve health and prevent diseases . The effects of polyphenols from different sources of propolis on atherosclerotic lesions and inflammatory and angiogenic factors were investigated in P01130 gene ( LDLr-/- ) knockout mice . The animals received a cholesterol-enriched diet to induce the initial atherosclerotic lesions ( IALs ) or advanced atherosclerotic lesions ( AALs ) . The IAL or AAL animals were divided into three groups , each receiving polyphenols from either the green , red or brown propolis ( 250 mg/kg per day ) by gavage . After 4 weeks of polyphenol treatment , the animals were sacrificed and their blood was collected for lipid profile analysis . The atheromatous lesions at the aortic root were also analyzed for gene expression of inflammatory and angiogenic factors by quantitative real-time polymerase chain reaction and immunohistochemistry . All three polyphenol extracts improved the lipid profile and decreased the atherosclerotic lesion area in IAL animals . However , only polyphenols from the red propolis induced favorable changes in the lipid profiles and reduced the lesion areas in AAL mice . In IAL groups , VCAM , P13500 , FGF , PDGF , P15692 , PECAM and P14780 gene expression was down-regulated , while the metalloproteinase inhibitor P01033 gene was up-regulated by all polyphenol extracts . In contrast , for advanced lesions , only the polyphenols from red propolis induced the down-regulation of P16671 and the up-regulation of P09601 and P01033 when compared to polyphenols from the other two types of propolis . In conclusion , polyphenols from propolis , particularly red propolis , are able to reduce atherosclerotic lesions through mechanisms including the modulation of inflammatory and angiogenic factors . 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 . 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 . 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 . 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 . 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 . Potential anticancer agents . XXIII . 1 . Qualitative structure -- activity relationship in the " classical " antifolates area . In order to obtain " classical " antifolates with improved therapeutic index , 32 new DB00563 analogues were synthesized and studied . Their structure -- activity relationship analysis led to the following conclusions : a ) the replacement of glutamyl moiety with other amino acids led to compounds which are powerful inhibitors of P00374 but were devoid of activity against L1210 leukemia , b ) the phenylic nucleus substitution with methoxy groups afforded potent inhibitors of P00374 and also effective derivatives against experimental tumors , c ) the insertion of an extra amino acid between the phenylic ring and the terminal moiety proved to be an unfavorable event for the activity of such compounds , d ) the MTX-analogues with the peptidic side chain grafted at P10643 of the pteridine ring were ineffective against both P00374 and L1210 leukemia . From the investigated derivatives p [ 2,4-diamino-6-pteridinyl ) -methyl-N10-methyl ] aminobenzoyl-L-leucine 16 is twofold more potent as MTX by respect to P00374 and could be used in MTX-resistant ( by impaired transport ) cell lines being more hydrophobic . 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 . 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 . 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 gene expression profiling analysis in workers occupationally exposed to benzene . Benzene is an important industrial chemical and an environmental contaminant , but the pathogenesis of hematotoxicity induced by chronic occupational benzene exposure remains to be elucidated . To gain an insight into the molecular mechanisms and new biomarkers , microarray analysis was used to identify the differentially expressed mRNA critical for benzene hematotoxicity . RNA was extracted from four chronic benzene poisoning patients occupationally exposed to benzene , three benzene-exposed workers without clinical symptoms and three health controls without benzene exposure and mRNA expression profiling was performed using Gene Chip Human Gene 2.0ST Arrays . Analysis of mRNA expression profiles were conducted to identify key genes , biological processes and pathways by the series test of cluster ( P52823 ) , P52823 -Gene Ontology analysis ( P52823 -GO ) , pathway analysis and Signal-net . PRINCIPAL FINDINGS : 1 ) 1661 differentially expressed mRNAs were identified and assigned to sixteen model profiles . 2 ) Profiles 14 , 10 , 11 , 1 and 15 are the predominant expression profiles which were involved in immune response , inflammatory response , chemotaxis , defense response , anti-apoptosis and signal transduction . 3 ) The importance of immune response at benzene hematotoxicity is highlighted by several immune-related signaling pathways such as B/T cell receptor signaling pathway , acute myeloid leukemia , hematopoietic cell lineage and natural killer cell mediated cytotoxicity . 4 ) Signet analysis showed that P27986 , P48736 , O00459 , P08754 , P01116 , P01111 , P19838 , P28067 , and P28068 were key genes involved in benzene hematotoxicity . These genes might be new biomarkers for the prevention and early diagnosis of benzene poisoning . This is a preliminary study that paves the way to further functional study to understand the underlying regulatory mechanisms . 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 . Development of a highly sensitive chemical method for detecting alpha2 --> 8-linked oligo/polysialic acid residues in glycoproteins blotted on the membrane . A highly sensitive chemical method for detecting alpha2 --> 8-linked oligo/polysialic acid ( oligo/polySia ) chains was developed , including ( i ) periodate oxidation , reduction with sodium borohydride , and subsequent acid hydrolysis , giving rise to P10643 analogues and intact P02748 compounds from nonreducing terminal and internal sialic acid residues , respectively ; ( ii ) fluorescent labeling of these P10643 and P02748 compounds with 1,2-diamino-4,5-methylenedioxybenzene ( P28068 ) ; and ( iii ) quantitation of theseDMB derivatives on fluorometric high-performance liquid chromatography . As little as 1 ng of internal sialic acid residues of oligo/polySia chains , the existence of which indicates the presence of oligo/polySia structure , was detectable by this method . This fluorometric P10643 / P02748 analysis was successfully applied to glycoproteins blotted onto a slit of polyvinylidene fluoride membranes and suggested the presence of some novel oligoSia-containing glycoproteins in pig embryonic brains . Gender disparity in LDL-induced cardiovascular damage and the protective role of estrogens against electronegative LDL . BACKGROUND : Increased levels of the most electronegative type of LDL , Q15004 , have been observed in the plasma of patients with metabolic syndrome ( MetS ) and ST-segment elevation myocardial infarction and can induce endothelial dysfunction . Because men have a higher predisposition to developing coronary artery disease than do premenopausal women , we hypothesized that LDL electronegativity is increased in men and promotes endothelial damage . METHODS : Q15004 levels were compared between middle-aged men and age-matched , premenopausal women with or without MetS . We further studied the effects of gender-influenced LDL electronegativity on aortic cellular senescence and DNA damage in leptin receptor-deficient ( db/db ) mice by using senescence-associated-β-galactosidase and γ P16104 staining , respectively . We also studied the protective effects of 17β-estradiol and genistein against electronegative LDL-induced senescence in cultured bovine aortic endothelial cells ( BAECs ) . RESULTS : Q15004 levels were higher in MetS patients than in healthy subjects ( P < 0.001 ) , particularly in men ( P = 0.001 ) . LDL isolated from male db/db mice was more electronegative than that from male or female wild-type mice . In addition , LDL from male db/db mice contained abundantly more apolipoprotein CIII and induced more BAEC senescence than did female db/db or wild-type LDL . In the aortas of db/db mice but not wild-type mice , we observed cellular senescence and DNA damage , and the effect was more significant in male than in female db/db mice . Pretreatment with 17β-estradiol or genistein inhibited BAEC senescence induced by male or female db/db LDL and downregulated the expression of lectin-like oxidized P01130 -1 and tumor necrosis factor-alpha protein . CONCLUSION : The gender dichotomy of LDL-induced cardiovascular damage may underlie the increased propensity to coronary artery disease in men . 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 . [ cDNA microarray and cluster analysis to identify the significance of immune genes associated with benzene poisoning ] . OBJECTIVE : To delineate the immune regulatory pathway of benzene poisoning by using gene expression profile analysis . METHODS : Peripheral white blood cell gene expression profile of 7 benzene poisoning patients , including one aplastic anemia , was determined by microarray . Seven chips from normal workers were served as controls . Cluster analysis of gene expression profile was performed . Differentially expressed immune genes associated with benzene poisoning were determined . RESULTS : Among the 2 779 target genes , 38 genes differentially expressed were identified , including 10 up-regulated genes such as P13987 , TRA@ , MCP etc , and 14 down-regulated genes such as P28068 , HLA-DQA1 , P04440 , P05107 , P27918 etc . Cluster analysis showed that the expression profiles of 38 genes were associated with benzene poisoning . CONCLUSION : Differentially expressed immune genes may play an important role in the pathogenesis of benzene poisoning . 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 . Two-dimensional liquid crystalline growth within a phase-field-crystal model . By using a two-dimensional phase-field-crystal ( P27918 ) model , the liquid crystalline growth of the plastic triangular phase is simulated with emphasis on crystal shape and topological defect formation . The equilibrium shape of a plastic triangular crystal ( PTC ) grown from an isotropic phase is compared with that grown from a columnar or smectic-A ( Q13216 ) phase . While the shape of a PTC nucleus in the isotropic phase is almost identical to that of the classical P27918 model , the shape of a PTC nucleus in Q13216 is affected by the orientation of stripes in the Q13216 phase , and irregular hexagonal , elliptical , octagonal , and rectangular shapes are obtained . Concerning the dynamics of the growth process , we analyze the topological structure of the nematic order , which starts from nucleation of +1/2 and -1/2 disclination pairs at the PTC growth front and evolves into hexagonal cells consisting of +1 vortices surrounded by six satellite -1/2 disclinations . It is found that the orientational and the positional order do not evolve simultaneously ; the orientational order evolves behind the positional order , leading to a large transition zone , which can span over several lattice spacings . 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 . Selective targeting of the repressive transcription factors P25490 and cMyc to disrupt quiescent human immunodeficiency viruses . Quiescent HIV-1 infection of resting P01730 (+) T cells is an obstacle to eradication of HIV-1 infection . These reservoirs are maintained , in part , by repressive complexes that bind to the HIV-1 long terminal repeat ( LTR ) and recruit histone deacetylases (HDACs). cMyc and P25490 are two transcription factors that are recruited as part of well-described , distinct complexes to the HIV-1 LTR and in turn recruit HDACs . In prior studies , depletion of single factors that recruit Q13547 in various cell lines was sufficient to upregulate LTR activity . We used short hairpin RNAs ( shRNAs ) to test the effect of targeted disruption of a single transcription factor on quiescent proviruses in T cell lines . In this study , we found that depletion of P25490 significantly increases mRNA and protein expression from the HIV-1 promoter in some contexts , but does not affect Q13547 , Q92769 , O15379 , or acetylated histone 3 occupancy of the HIV-1 LTR . Conversely , depletion of cMyc or cMyc and P25490 does not significantly alter the level of transcription from the LTR or affect recruitment of HDACs to the HIV-1 LTR . Furthermore , global inhibition of HDACs with the HDAC inhibitor suberoylanilide hydroxamic acid ( DB02546 ) enhanced the increase in LTR transcription in cells that were depleted of P25490 .These findings show that despite prior isolated findings , redundancy in repressors of HIV-1 LTR expression will require selective targeting of multiple restrictive mechanisms to comprehensively induce the escape of quiescent proviruses from latency . 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 . Evaluation of the aortic root by Q9BWK5 : insights from patients with homozygous familial hypercholesterolemia . BACKGROUND : In homozygous familial hypercholesterolemia ( HFH ) , the aortic root is prone to develop atherosclerotic plaque at an early age . However , the aortic wall and plaque have not yet been assessed in this condition by Q9BWK5 . We evaluated the aortic root by use of Q9BWK5 in 17 HFH patients and 12 normal control subjects in a prospective , blinded , controlled study . METHODS AND RESULTS : Morphological assessment of the aortic root was done with spin-echo and gradient-echo Q9BWK5 scanning . Comparisons were made with a number of measures of disease severity , including cholesterol-year score , calcium score on electron-beam CT ( EBCT ) , and size of Achilles tendon xanthomas . Atherosclerotic plaque , visible on fat-suppressed images but never on water-suppressed images , was present in 9 HFH patients ( 53 % ) . Supravalvular aortic stenosis was present in 7 patients with HFH ( 41 % ) . Maximum supravalvular aortic wall thickness was significantly greater and OD and lumen cross-sectional area ( Q13216 ) were smaller in patients than in control subjects ( P=0.006 , 0.0005 , and 0.06 , respectively ) . Maximum wall thickness was associated with a greater calcium score on electron-beam CT ( P=0.02 ) . Although the cumulative exposure of the aortic root to cholesterol ( the cholesterol-year score ) was significantly correlated with the Achilles tendon Q13216 and vascular calcification , this score did not correlate with the wall thickness or aortic Q13216 . CONCLUSIONS : This study not only demonstrates the utility of Q9BWK5 for detecting and characterizing aortic root atherosclerotic plaque and supravalvular aortic stenosis in HFH patients but also suggests that the P01130 plays a direct or indirect role in aortic mural development and vascular growth . 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 . 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 . 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 . 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 . P78380 expressed in cultured rat chondrocytes mediates oxidized LDL-induced cell death-possible role of dephosphorylation of Akt . The oxidative changes of lipids in cartilage proceed with ageing and with the grade of osteoarthritis . To clarify the role of oxidatively modified lipids in articular cartilage in osteoarthritis , here , we investigated lectin-like oxidized P01130 ( P78380 ) in rat cultured articular chondrocytes . P78380 expression was detectable in basal culture condition and enhanced by the treatment of oxidized LDL and interleukin-1beta . DiI-labeled oxidized LDL was bound and ingested by chondrocytes via P78380 . Oxidized LDL dose-dependently reduced chondrocyte viability , inducing non-apoptotic cell death , which was again suppressed by anti- P78380 antibody treatment . Oxidized LDL reduced the amount of phosphorylated Akt , a substrate of P19957 kinase via P78380 . Consistently , the P19957 kinase inhibitor , LY294002 , decreased cell viability dose-dependently , and the P19957 kinase activator , P05019 , reversed the effect of oxidized LDL on the cell death . P78380 might be involved in the pathogenesis of osteoarthritis , inducing chondrocyte death through P19957 kinase/Akt pathway . 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 . 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 . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used .

MH_train_1123

MH_train_1123

MH_train_1123

interacts_with DB00091?

multiple_choice

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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . [ 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 . [ 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 . 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 . 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 . 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 . Association of novel SNPs in the candidate genes affecting caprine milk fatty acids related to human health . In the present investigation , 618 milk samples of Sirohi breed of goat were collected , and analyzed for conjugated linoleic acid ( DB01211 , C18:2 ) and other fatty acids . The DB01211 in studied goat milk samples was 4.87 mg/g of milk fat and C18:2 cis-9 , trans-11 contributes 2.9 mg/g of milk fat and trans10 cis12 contributes 0.82 mg/g of milk fat . The saturated fatty acids in the milk accounted for 69.55 % and unsaturated fatty acid accounted for 28.50 % . The unsaturated fatty acid was constituted by monounsaturated fatty acid ( 24.57 % ) and polyunsaturated fatty acids ( 3.96 % . ) . The major contribution ( 45.56 % ) in total fatty acid was of C12:0 , C14:0 and C16:0 . C18:0 and short chain ones ( C4:0 , P13671 :0 , Q99618 :0 , and Q99622 :0 ) have a neutral or cholesterol-decreasing effect . The DNA sequence analysis of the genes ( O75907 , SCAP , P37231 , OLR , P05413 and PRL ) in a random panel of 8 Sirohi goats revealed 38 SNPs across the targeted regions . Out of the studied SNPs ( 38 ) across these genes , 22 SNPs had significant effect on one or a group of fatty acids including DB01211 . The genotypes at these loci showed significant differences in the least square means of a particular fatty acid or a group of fatty acids including DB01211 and its isomers . Genotypes associated with lipid metabolism contribute to differences in serum lipid profile of GH-deficient adults before and after GH replacement therapy . OBJECTIVE : GH deficiency ( GHD ) in adults is associated with an altered serum lipid profile that responds to GH replacement therapy ( GHRT ) . This study evaluated the influence of polymorphisms in genes related to lipid metabolism on serum lipid profile before and after 1 year of GHRT in adults . DESIGN AND METHODS : In 318 GHD patients , total cholesterol ( TC ) serum concentrations , LDL-C , HDL-C , and triglycerides ( TG ) were assessed . Using a candidate gene approach , 20 single nucleotide polymorphisms ( SNPs ) were genotyped . GH dose was individually titrated to obtain normal serum IGF1 concentrations . RESULTS : At baseline , the minor alleles of cholesteryl ester transfer protein ( P11597 ) gene SNPs rs708272 and rs1800775 were associated with higher serum TC and apolipoprotein E ( P02649 ) gene SNP rs7412 with lower TC concentrations ; P11597 SNPs rs708272 , rs1800775 , and rs3764261 and apolipoprotein B ( P04114 ) gene SNP rs693 with higher serum HDL-C ; P02649 SNP rs7412 , peroxisome proliferator-activated receptor gamma ( P37231 ) gene SNP rs10865710 with lower LDL-C , and P11597 SNP rs1800775 with higher LDL-C ; and P02649 /C1/C4/ P06681 cluster SNP rs35136575 with lower serum TG . After treatment , P04114 SNP rs676210 GG genotype was associated with larger reductions in TC and LDL-C and P37231 SNP rs10865710 CC genotype with greater TC reduction . All associations remained significant when adjusted for age , sex , and BMI . CONCLUSIONS : In GHD adults , multiple SNPs in genes related to lipid metabolism contributed to individual differences in baseline serum lipid profile . The GH treatment response in TC and LDL-C was influenced by polymorphisms in the P04114 and P37231 genes . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Investigation of a potential protective mechanism against heparin-induced thrombocytopenia in patients on chronic intermittent hemodialysis . BACKGROUND : DB01109 -induced thrombocytopenia ( HIT ) develops as a result of platelet ( Q02083 ) activation by anti-platelet factor 4 ( P02776 ) /heparin complex antibodies . Despite repeated exposure to heparin , patients undergoing chronic intermittent hemodialysis ( HD ) rarely develop HIT . We investigated the possibility that HD decreases/removes P02776 from Q02083 surfaces and/or plasma , thereby disfavoring immune complex formation as a mechanism of protection against HIT . MATERIALS AND METHODS : We enrolled 20 patients undergoing chronic HD at the Penn Presbyterian Medical Center . Blood samples were drawn before , during and after treatment in the presence and absence of heparin . P02776 , anti- P02776 /heparin antibody , heparin , and P16109 levels were measured . RESULTS : No patients demonstrated clinical symptoms of HIT . Q02083 surface P02776 levels decreased and plasma P02776 levels increased concurrently with the increase in plasma heparin concentration . In the absence of heparin , Q02083 surface and plasma P02776 levels were unchanged . Anti- P02776 /heparin antibodies , which were non-functional by the serotonin release assay , were detectable in 8 patients . Q02083 surface P16109 levels did not change during treatment . CONCLUSIONS : Removal of Q02083 surface and/or plasma P02776 as a mechanism of protection against HIT in patients undergoing HD is not supported by the results of our study , although the transient decrease in Q02083 surface P02776 in the presence of large amounts of heparin remains a candidate mechanism . The small sample size , single type of dialyzer membrane , and early sampling time points may have led to the inability to detect changes in P02776 levels . Future studies should explore other potential protective mechanisms . 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 . APRIL and Q9Y275 proteins increase proliferation of human adipose-derived stem cells through activation of Erk1/2 Q96HU1 kinase . Human adipose-derived stem cells ( Q9ULZ3 ) are DB05914 with reduced immunogenicity and the ability to modulate immune responses . APRIL and Q9Y275 proteins are overexpressed in inflammatory and autoimmune diseases for which allogeneic Q9ULZ3 therapy is currently under clinical investigation . Modification of Q9ULZ3 properties by the tissue microenvironment could be a critical factor in patient outcome and is still not well understood . Our aim was to characterize the APRIL/ Q9Y275 system in Q9ULZ3 by analyzing the ligand and receptor expression patterns , the effects mediated by APRIL and Q9Y275 on Q9ULZ3 , and the underlying signaling . We found that Q9ULZ3 express the tumor necrosis factor proteins APRIL ( a proliferation-inducing ligand ) and Q9Y275 ( B cell-activator factor ) as well as their receptors O14836 ( transmembrane activator and calcium-modulator and cyclophilin ligand interactor ) , Q02223 ( B cell maturation antigen ) and the Q9Y275 -specific receptor ( Q96RJ3 ) . APRIL and Q9Y275 secretion was differentially enhanced by P48061 and interferon ( IFN ) -γ , implicated in Q9ULZ3 -mediated migration and immunosuppression , respectively . In addition , APRIL and Q9Y275 induced rapid phosphorylation of extracellular signal-regulated kinases 1/2 ( P27361 /2 ) and Akt kinases and promoted an increase in Q9ULZ3 proliferation , without affecting the immunosuppressive capacity of these cells . The use of specific chemical inhibitors indicated that the PI3K transduction pathway is involved in Q9ULZ3 basal growth and that APRIL- and Q9Y275 -mediated effects are P29323 -dependent . These results provide new information about the molecular mechanisms that underlie APRIL and Q9Y275 secretion and signaling in Q9ULZ3 , and are of special relevance for the use of allogeneic Q9ULZ3 as therapeutic tools . 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 . Long pentraxin-3 as an epithelial-stromal fibroblast growth factor-targeting inhibitor in prostate cancer . Fibroblast growth factors ( FGFs ) exert autocrine/paracrine functions in prostate cancer by stimulating angiogenesis and tumour growth . Here dihydrotestosterone ( DB02901 ) up-regulates P09038 and FGF8b production in murine TRAMP- P06681 prostate cancer cells , activating a FGF-dependent autocrine loop of stimulation . The soluble pattern recognition receptor long pentraxin-3 ( PTX3 ) acts as a natural FGF antagonist that binds P09038 and FGF8b via its N-terminal domain . We demonstrate that recombinant PTX3 protein and the PTX3-derived pentapeptide Ac-ARPCA-NH2 abolish the mitogenic response of murine TRAMP- P06681 cells and human LNCaP prostate cancer cells to DB02901 and FGFs . Also , PTX3 hampers the angiogenic activity of DB02901 -activated TRAMP- P06681 cells on the chick embryo chorioallantoic membrane ( P62158 ) . Accordingly , human PTX3 overexpression inhibits the mitogenic activity exerted by DB02901 or FGFs on hPTX3_TRAMP- P06681 cell transfectants and their angiogenic activity . Also , hPTX3_TRAMP- P06681 cells show a dramatic decrease of their angiogenic and tumourigenic potential when grafted in syngeneic or immunodeficient athymic male mice . A similar inhibitory effect is observed when TRAMP- P06681 cells overexpress only the FGF-binding N-terminal PTX3 domain . In keeping with the anti-tumour activity of PTX3 in experimental prostate cancer , immunohistochemical analysis of prostate needle biopsies from primary prostate adenocarcinoma patients shows that parenchymal PTX3 expression , abundant in basal cells of normal glands , is lost in high-grade prostatic intraepithelial neoplasia and in invasive tumour areas . These results identify PTX3 as a potent FGF antagonist endowed with anti-angiogenic and anti-neoplastic activity in prostate 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 ) 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 . 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 . DB00472 -induced proliferation and differentiation of neural progenitor cells isolated from rat postnatal cerebellum . Previous studies have shown that the serotonin-reuptake inhibitor ( SSRI ) fluoxetine affects neural progenitors derived from postnatal cerebellum or adult hippocampus and stimulates their proliferation . In the human cerebellum , the proliferation of cerebellar granule cells ( CGC ) continues until the 11th postnatal month and could be influenced in infants by breastfeeding-delivered SSRIs . Current information about fluoxetine effects on postnatal cerebellar neural progenitors is limited . Here we report the characterization of fluoxetine actions on rat postnatal cerebellar neural progenitors . RT-PCR and immunostaining revealed the expression of serotonin transporter ( P31645 ) , 5HT(1A) receptors , tryptophan hydroxylase ( P17752 ) , and serotonin ( 5HT ) . Protracted in vitro fluoxetine treatment increased cell proliferation and differentiation . The proliferative effects of fluoxetine , 5HT , and the selective agonist of 5HT(1A) receptors trans-8-hydroxy-2-(N-n-propyl-N-3'-iodo-2'-propenyl)aminotetralin ( 8-OH-PIPAT ) were abolished by the selective antagonist of 5HT(1A) receptors , N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohexanecarboxamide trihydrochloride ( WAY-100635 ) . Furthermore , fluoxetine-induced activation of both the DB02527 -response element-binding ( CREB ) protein and extracellular signal-regulated protein kinases ( P27361 /2 ) , which was abolished by the selective inhibitor of Q96HU1 kinase kinase ( MEK ) 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene ( U0126 ) , and increased cyclin D1 expression . All these effects were prevented by WAY-100635 . Collectively , our results demonstrate that rat postnatal cerebellum contains neural progenitors capable of proliferating and differentiating in response to fluoxetine exposure , possibly through the activation of 5HT(1A) receptors . The relevance of these findings for possible SSRI effects on the developing postnatal/infant human cerebellum should be explored . 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 . 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 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 . 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 . 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 . 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 . 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 . [ 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 . 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 . 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 . Molecular profiling to identify molecular mechanism in esophageal cancer with familial clustering . To identify the genes and molecular functional pathways involved in esophageal cancer , we analyzed the gene expression profile of esophageal tumor tissue from patients having family history of esophageal cancer by cDNA microarray . Three hundred and fifty differentially expressed genes ( 26 up-regulated and 324 down-regulated ) were identified . Genes involved in humoral immune response ( P02776 ) , extracellular matrix organization ( P53420 ) , metabolism of xenobiotics ( P07099 ) , TGF-beta signaling ( Q15797 ) and calcium signaling pathways ( P21796 ) were down-regulated and genes involved in regulation of actin cytoskeleton ( O00401 ) , neuroactive ligand receptor interaction ( Q14832 ) , Toll-like receptor ( P08571 ) , B-cell receptor ( P13164 ) and insulin signaling pathways ( Q12778 ) were up-regulated . Validation of differential expression of subset of genes by QRT-PCR and tissue microarray in familial and non-familial cases showed no significant difference in expression of these genes in two groups suggesting familial clustering occurs as result of sharing of common environmental factors . Gene expression profiling of clinical specimens from well characterized populations that have familial clustering of cancer identified molecular mechanism associated with progression of esophageal cancer . 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 . 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 . 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 . P62937 may contribute to the inflammatory processes in rheumatoid arthritis through induction of matrix degrading enzymes and inflammatory cytokines from macrophages . P62937 ( CypA ) levels increase in the sera and synovial fluids of rheumatoid arthritis ( RA ) patients , but the cell types expressing CypA and the function of CypA in the pathogenesis of RA are not known yet . Immunohistochemistry analyses revealed high level CypA staining in the macrophages in the lining layers of human RA and osteoarthritis synovium . Low level CypA staining was also detected in endothelial cells , lymphocytes , and smooth muscle cells in RA synovium . Further investigation of the CypA function using monocyte/macrophage cell lines revealed that CypA induced expression of cytokine/chemokines such as P01375 , P10145 , P13500 , and IL-1beta and matrix metalloproteinase ( MMP ) -9 through a pathway that is dependent on NFkappaB activation . Furthermore , P14780 staining pattern overlapped with that of CypA in both RA and OA synovium . Our data suggest that CypA may stimulate macrophages to degrade joint cartilage via P14780 expression and promote inflammation via pro-inflammatory cytokine secretion . 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 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 . 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 . 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 . 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 . 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 . 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) . 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 . 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 . Expression of angiotensin I-converting enzymes and bradykinin B2 receptors in mouse inner medullary-collecting duct cells . We described in mouse inner medullary-collecting duct cells ( mIMCD-3 ) the somatic and the N-domain P12821 synthesis and its interaction with the kallikrein-kinin system co-localized in the same cells . We purified two P12821 forms from culture medium , M1 ( 130 kDa ) and M2 ( N-domain , 60 kDa ) , and cellular lysate , C1 ( 130 kDa ) and P06681 ( N-domain , 60 kDa ) . DB01197 and enalaprilat inhibited the purified enzymes . The immunofluorescence studies indicated that P12821 is present in the membrane , cytoplasm and in the cell nucleus . Kinin B1 and B2 receptors were detected by immunofluorescence and showed to be activated by BK and DesR9 BK , increasing the acidification rate which was enhanced in the presence of enalaprilat . The presence of secreted and intracellular P12821 in mIMCD-3 confirmed the hypothesis previously proposed by our group for a new site of P12821 secretion in the collecting duct . Prevention of high-mobility group box 1-mediated early loss of transplanted mouse islets in the liver by antithrombin III . BACKGROUND : The low efficiency of pancreatic islet transplantation mainly because of the early loss of transplanted islets hampers its clinical application . Previously , we have shown in mice that the early loss of transplanted islets in the liver is caused by innate immune rejection in concert with dendritic cells , natural killer T cells , and neutrophils to produce interferon ( IFN ) -γ , which is triggered by high-mobility group box 1 ( P09429 ) released from transplanted islets . We herein determined whether the P09429 -mediated early loss of transplanted mouse islets is prevented by antithrombin ( P01008 ) . METHODS : The effect of P01008 on in vitro and in vivo P09429 -stimulated IFN-γ production of hepatic mononuclear cells was examined . Then , the effect of P01008 on amelioration of hyperglycemia in streptozotocin-induced diabetic mice receiving 200 syngeneic islets from a single donor was determined . RESULTS : In vitro and in vivo IFN-γ production of mononuclear cells in the liver of mice in response to P09429 was suppressed by P01008 . Hyperglycemia of streptozotocin-induced diabetic mice receiving 200 syngeneic islets into the liver from a single donor was ameliorated with down-regulation of IFN-γ production of natural killer T cells and neutrophils in the liver when P01008 but not vehicle was administered once at the time of islet transplantation . The favorable effect of P01008 was similarly achieved in mice receiving islet allografts when rejection was prevented with anti- P01730 antibody treatment . CONCLUSIONS : These findings demonstrate that P01008 prevents P09429 -mediated early loss of transplanted islets caused by innate immune rejection , suggesting a potential application of P01008 to improve efficiency of clinical islet transplantation .

MH_train_1124

MH_train_1124

MH_train_1124

interacts_with DB00831?

multiple_choice

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 . 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 . P06850 -induced adrenocorticotropin hormone release and synthesis is blocked by incorporation of the inhibitor of cyclic AMP-dependent protein kinase into anterior pituitary tumor cells by liposomes . P06850 ( CRF ) is the most potent and effective natural stimulant of corticotropin ( DB01285 ) secretion . In a tumor cell line of the mouse anterior pituitary ( AtT-20/D16-16 ) consisting of a homogeneous population of corticotrophs , CRF is known to increase adenylate cyclase and DB02527 -dependent protein kinase activities as well as to release DB01285 . To determine whether activation of DB02527 -dependent protein kinase is essential for CRF to evoke the secretion of DB01285 , an inhibitor ( PKI ) of this kinase was inserted into AtT-20 cells . This was accomplished by first encapsulating PKI into liposomes and then covalently coupling them to protein A for binding to antibodies directed against an AtT-20 cell surface antigen , N- P62158 ( neural cell adhesion molecule ) . The binding of the liposomes to the anti-N- P62158 antibodies led to the internalization of the PKI into the tumor cells . The PKI treatment greatly attenuated CRF-stimulated DB01285 release as well as the secretory response to beta-adrenergic agonists . However , DB01285 release in response to caerulein , an agonist of cholecystokinin 8 receptors , was not altered by the PKI treatment . CRF treatment also increased the levels of mRNA for proopiomelanocortin ( P01189 ) , the precursor for DB01285 in AtT-20 cells . Application of liposomes containing PKI to AtT-20 cells blocked the ability of CRF and 8-bromo- DB02527 , but not phorbol ester , to increase P01189 mRNA levels . The results revealed an essential role for DB02527 in mediating the effect of CRF on DB01285 release and P01189 gene expression . 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 . 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 . 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 . 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 . [ 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 . Resistance of P04626 /neu-overexpressing tumor targets to lymphokine-activated-killer-cell-mediated lysis : evidence for deficiency of binding and post-binding events . P04626 /neu-overexpressing tumor cell lines are relatively resistant to lymphokine-activated killer ( Q96QP1 ) cell cytotoxicity when compared to P04626 /neu-nonexpressing lines . P04626 /neu+ targets were also resistant to binding by Q96QP1 large granular lymphocytes ( LGL ) as shown by visualization at the single-cell level , a target monolayer binding assay and in " cold " target inhibition experiments . P04626 /neu+ Q96QP1 -resistant ovarian cell lines demonstrated an absence of P05362 expression while expression of LFA-3 , N- P62158 , laminin and beta 1 integrins was comparable to that of P04626 /neu- targets . In contrast , the P04626 /neu+ breast cell line , SKBR-3 , which was also resistant to lysis and binding by Q96QP1 LGL , demonstrated normal expression of P05362 . Anti- P05362 antibodies blocked binding and lysis of P04626 /neu- carcinoma targets by Q96QP1 cells , further supporting the notion that lack of P05362 expression on P04626 /neu+ cells contributes to their resistance . The modest binding and lysis of P04626 /neu+ targets by Q96QP1 cells was significantly inhibited by anti-LFA-1 antibodies , suggesting the existence of another counter-receptor for LFA-1 on P04626 /neu+ targets . The following also supported deficiencies in post-binding events when P04626 /neu+ cells resisted the lytic activity of Q96QP1 cells : ( a ) when the relative resistance to effector cell binding was overcome by exogenous lectin . P04626 /neu+ cell lines were still resistant to Q96QP1 cytolysis , and ( b ) P04626 /neu+ targets were resistant to perforin-containing granule extracts obtained from the CTLL-R8 cytotoxic lymphocyte cell line . These results indicate that deficiency in effector binding as well as post-binding events contributes to the resistance of P04626 /neu-overexpressing tumor targets to Q96QP1 -cell-mediated lysis . P29323 is an anti-inflammatory signal that suppresses expression of NF-kappaB-dependent inflammatory genes by inhibiting IKK activity in endothelial cells . Unveiling of endothelial nuclear factor-kappaB ( NF-kappaB ) activation is pivotal for understanding the inflammatory reaction and the pathogenesis of inflammatory vascular diseases . We here report the novel function of extracellular signal-related kinase ( P29323 ) in controlling endothelial NF-kappaB activation and inflammatory responses . In human endothelial cells , vascular endothelial growth factor ( P15692 ) induced NF-kappaB-dependent transcription of cell adhesion molecules ( CAMs ) and monocyte adhesion . These effects were prominently enhanced by either pretreatment with the MEK inhibitors , PD98059 and U0126 or overexpression of a dominant negative form of MEK , but blocked by a wild type P29323 . Consistently , inhibition of P29323 significantly increased O15111 ( IKK ) activity , P25963 phosphorylation , and nuclear translocation of NF-kappaB induced by P15692 , whereas overexpression of P29323 resulted in the loss of these responses to P15692 . Using two PKC inhibitors has demonstrated that P15692 concomitantly stimulates IKK and its negative regulatory signal P29323 through PKC that lies downstream of P35968 /Flk-1 . Strikingly , elevation of P29323 in endothelial cells markedly inhibited P62158 expression and NF-kappaB activation as well as monocyte adhesion induced by IL-1beta and P01375 . The data collectively suggest that P29323 serves as an anti-inflammatory signal that suppresses expression of NF-kappaB-dependent inflammatory genes by inhibiting IKK activity in endothelial cells . Measuring the existence of P29323 activity in vascular endothelial cells may be useful for predicting the feasibility and potency of inflammatory reactions in the vasculature . Inhibitory effect of chlorpromazine on O14788 -induced osteoclastogenesis in mouse bone marrow cells . Chlorpromazine ( CPZ ) , the first widely used phenothiazine tranquilizer , is shown to inhibit the action of intracellular calmodulin ( P62158 ) and bone resorption in vivo and in vitro . In this study , CPZ ( 0.63 - 10 µM ) dose-dependently inhibited the formation of tartrate-resistant acid phosphatase ( TRAP ) staining-positive osteoclast-like cells in mouse bone marrow cells ( BMCs ) treated with 1α,25(OH)(2)D(3) ( 10 nM ) or soluble receptor activator of nuclear factor-κB ligand ( s- O14788 ) ( 20 ng/ml ) . Expressions of mRNA for the nuclear factor of activated T-cells c1 ( O95644 ) , a key regulator of osteoclast differentiation ; dendritic cell-specific transmembrane protein ( Q9H295 ) , an essential protein for cell-cell fusion ; and characteristic markers of osteoclasts such as TRAP , cathepsin K , carbonic anhydrase II , and calcitonin receptor in BMCs were up-regulated by s- O14788 and decreased by the addition of CPZ ( 5 µM ) or the selective P62158 antagonist W7 , but not the inactive analog W5 . The general P62158 kinase ( CaMK ) inhibitor KN-93 and P62158 -dependent phosphatase calcineurin inhibitor FK-506 also inhibited s- O14788 -induced osteoclastogenesis . Phenothiazines such as CPZ , trifluoperazine ( TFPZ ) , and promethazine ( PMZ ) inhibited s- O14788 -induced osteoclast-like cell formation in mouse BMCs . Osteoclastogenesis inhibitory effects decreased in the order of TFPZ , CPZ , PMZ , depending on their anti- P62158 potency . These findings suggest that CPZ inhibits O14788 -induced osteoclastogenesis by its anti- P62158 action .

MH_train_1125

MH_train_1125

MH_train_1125

interacts_with DB00188?

multiple_choice

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 . 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 . 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 . 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 . Rapid immunomodulation by rosuvastatin in patients with acute coronary syndrome . AIMS : P04035 inhibitors ( statins ) reduce cardiovascular mortality and morbidity in patients with stable coronary artery disease as well as acute coronary syndrome ( ACS ) . It is unclear how rapidly the beneficial effects of statins occur in patients with ACS and whether these drug properties are related to lipid lowering . METHODS AND RESULTS : Patients with troponin-positive ACS ( n=35 ) were randomized to 20 mg/day rosuvastatin therapy or to placebo treatment . Anti-inflammatory effects of rosuvastatin measured by lymphocyte intracellular cytokine production were taken before initiation of treatment and on days 1 , 3 , and 42 . Compared with placebo , rosuvastatin treatment significantly reduced plasma concentrations of pro-inflammatory cytokines P01375 and P01579 at 72 h . DB01098 also induced a rapid and significant reduction of P01375 and P01579 production in stimulated T-lymphocytes at 72 h . When compared with placebo , rosuvastatin inhibited the Th-1-immune response measured at 72 h . CONCLUSION : DB01098 exerts rapid immunomodulatory effects on the level of T-cell activation in patients with ACS . 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 % ) . 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 . 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 . 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 . No evidence of mutations of the P28074 ( beta-5 subunit of proteasome ) in a case of myeloma with clinical resistance to DB00188 . 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 . 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 . 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 . TLR-4 signalling pathway : MyD88 independent pathway up-regulation in chicken breeds upon LPS treatment . Toll-like receptors ( TLRs ) that sense the microbial pathogens are important components of host immune system . TLRs play key roles in the innate defence mechanism against pathogens , in the development of adaptive immunity , and are possibly the major determinants of the susceptibility to infections . To study the resistance pattern in different breeds of chicken , a comprehensive understanding of O00206 signalling pathways is required . We investigated the TLR-4 pathway regulated gene expressions in PBMCs of chicken breeds of Broiler ( Cobb ) , Aseel , Dahlem Red and Ghagus upon LPS treatment using Quantitative RT-PCR approach . Several genes were found to be up regulated in both TLR-induced MyD88-dependent and MyD88-independent pathways . These genes include O00206 ( O00206 ) , MyD88 ( Myeloid differentiation primary response gene 88 ) , Q9Y4K3 ( P01375 receptor associated factor 6 ) , Q8IUC6 ( TIR domain containing adapter inducing interferon beta ) , the transcription factors NFkB ( Nuclear factor kappa B ) , Q92985 ( Q92985 ) and IFN β ( P01574 ) . We have also studied inflammatory cytokines such as P60568 , P05231 , P10145 , IL1 β and P01375 α to further understand the downstream signalling of O00206 pathway . These results showed that higher expression of TLR signalling activation via both MyD88-dependent and Q8IUC6 -dependent pathways are more beneficial to chicken mononuclear cells mediated innate immunity . We observed Q8IUC6 dependent pathway in Aseel and Ghagus breeds . Our results are in concurrent with general observation that Aseel breed is comparatively more resistant , Ghagus and broilers are moderately resistant and Dahlem Red is comparatively more susceptible to bacterial infections . DB00227 -induced proliferation inhibition and apoptosis in P13671 glial cells . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase is the rate-limiting enzyme in cholesterol biosynthesis . P04035 converts HMG- DB01992 to mevalonate , which is then converted into cholesterol or various isoprenoids through multiple enzymatic steps . In this study , we examined the cytotoxic effects of lovastatin , an P04035 inhibitor , in P13671 glial cells . DB00227 at concentrations higher than 10 microM suppressed cell proliferation and induced cell death , which were prevented completely by mevalonate ( 300 microM ) . The data from lactate dehydrogenase assay and fluorescence microscopic assay using Hoechst 33342 and propidium iodide showed that mevalonate at a concentration of 100 microM could prevent lovastatin-induced cell death , whereas it could not prevent lovastatin-induced inhibition of cell proliferation . These data suggest that the lovastatin-induced interruption of cell cycle transition was not sufficient to induce cell death in P13671 glial cells . In the presence of lovastatin at concentrations higher than 10 microM , DNA laddering , the typical finding of apoptosis , was identified . DB00227 -induced apoptosis was prevented by mevalonate ( 100 microM ) . Both cycloheximide ( 0.5 microgram/ml ) and actinomycin D ( 0.1 microgram/ml ) prevented lovastatin-induced DNA laddering . In this study , we demonstrated that the cytotoxic effects of lovastatin fall into two categories : suppression of cell growth and induction of apoptosis in P13671 glial cells . 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 . Receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) increases vascular permeability : impaired permeability and angiogenesis in P29474 -deficient mice . Receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) is emerging as an important regulator of vascular pathophysiology . Here , we demonstrate a novel role of O14788 as a vascular permeability factor and a critical role of endothelial nitric oxide synthase ( P29474 ) in O14788 -induced endothelial function . O14788 increased the vascular permeability and leukocyte infiltration in vivo and caused the breakdown of the blood-retinal barrier in wild-type mice but not in P29474 -deficient mice . In vitro , it increased endothelial permeability and reduced P33151 -facilitated endothelial cell-cell junctions in a NO-dependent manner . O14788 also led to the activation of Akt and P29474 and to NO production in endothelial cells ( ECs ) . These effects were suppressed by the inhibition of Q9Y4K3 , phosphoinositide 3'-kinase ( PI3K ) , Akt , or NOS by genetic or pharmacologic means . Inhibition of the Q9Y4K3 -mediated NO pathway reduced EC migration and capillary-like tube formation in response to O14788 . Moreover , the effects of O14788 on ECs sprouting from the aorta , and neovessel formation in both the mouse Matrigel plug assay and corneal micropocket assay , were impaired in P29474 -deficient mice . These results demonstrate that O14788 promotes vascular permeability and angiogenesis by stimulating P29474 by a Q9Y4K3 -PI3K-Akt-dependent mechanism . These properties may be relevant to the pathogenesis of angiogenesis-dependent and inflammatory vascular diseases . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 .

MH_train_1126

MH_train_1126

MH_train_1126

interacts_with DB00054?

multiple_choice

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 . Dexamethasone inhibits interleukin-1β-induced matrix metalloproteinase-9 expression in cochlear cells . OBJECTIVES : To investigate the effect of interleukin ( IL ) -1β on matrix metalloproteinase ( MMP ) -9 expression in cochlea and regulation of IL-1β-mediated P14780 expression by dexamethasone and the molecular and signaling mechanisms involved . METHODS : House ear institute-organ of Corti 1 ( HEI-OC1 ) cells were used and exposed to IL-1β with/without dexamethasone . P04150 antagonist , DB00834 , was used to see the role of dexamethasone . PD98059 ( an extracellular signal-regulated kinases [ ERKs ] inhibitor ) , SB203580 ( a p38 mitogen-activated protein kinases [ MAPK ] inhibitor ) , SP600125 ( a c-Jun N-terminal kinase [ JNK ] inhibitor ) were also used to see the role of MAPKs signaling pathway(s) in IL-1β-induced P14780 expression in HEI-OC1 cells . Reverse transcription-polymerase chain reaction and gelatin zymography were used to measure mRNA expression level of P14780 and activity of P14780 , respectively . RESULTS : Treatment with IL-1β-induced the expression of P14780 in a dose- and time-dependent manner . IL-1β ( 1 ng/mL ) -induced P14780 expression was inhibited by dexamethasone . Interestingly , p38 MAPK inhibitor , SB203580 , significantly inhibited IL-1β-induced P14780 mRNA and P14780 activity . However , inhibition of JNKs and ERKs had no effect on the IL-1β-induced P14780 expression . CONCLUSION : These results suggest that the pro-inflammatory cytokine IL-1β strongly induces P14780 expression via activation of p38 MAPK signaling pathway in HEI-OC1 cells and the induction was inhibited by dexamethasone . 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 . 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 . Hematopoietic differentiation of embryonic stem cells : an in vitro model to study gene regulation during megakaryocytopoiesis . We are interested in the regulation of the tissue specificity of the megakaryocyte-specific platelet glycoprotein IIb gene . The murine embryonic stem ( ES ) cells are able to differentiate into erythroid , mast and granulomonocytic cells in appropriate culture conditions . Our goal is to optimize the production of myeloid cells including megakaryocytes ( MKs ) by ES cells . We have found that coculture with MS-5 stromal cells and the presence of a cocktail of hematopoietic growth factors ( HGFs ) [ stem cell factor , interleukin 3 ( P08700 ) , P05231 , IL-11 , G- P04141 and erythropoietin ] had a high synergistic activity on differentiation of ES cells into pure and MK-containing myeloid colonies from day 12 embryoid bodies . P40225 increased the number of MKs only when added to the P14210 cocktail in the presence of MS-5 cells . Interestingly , many MKs exhibited a " hairy " appearance evocative of pseudopodial proplatelet formation . Expression of genes specific for the megakaryocytic lineage , P08514 , P02776 , mpl and P05106 , was detected by reverse transcriptase-polymerase chain reaction ( RT-PCR ) during differentiation of ES cells , and their relative time course was evaluated . This demonstrates that optimized culture conditions for the differentiation of ES cells into the MK lineage provide a useful tool for the study of the regulation of expression of genes during megakaryocytopoiesis . Genomic structure and chromosome location of the murine Q01064 phosphodiesterase gene . Cyclic nucleotide phosphodiesterases ( PDEs ) catalyze the hydrolysis of DB02527 and cGMP , thereby participating in regulation of the intracellular concentrations of these second messengers . The PDE1 family is defined by regulation of activity by calcium and calmodulin . We have cloned and characterized the mouse Q01064 gene , which encodes the 63-kDa calcium/calmodulin-dependent PDE ( P62158 -PDE ) , an isozyme that is expressed in the CNS in the olfactory tract , dentate gyrus , and striatum and may participate in learning , memory , and regulation of phosphorylation of Q9UD71 in dopaminergic neurons . We screened an I-129/SvJ mouse genomic library and identified exons 2-13 of the Q01064 gene that span 8.4 kb of genomic DNA . Exons range from 67 to 205 nucleotides and introns from 91 to 2250 nucleotides in length . Exon 1 was not present in the 3 kb of genomic DNA 5' to exon 2 in our clones . The mouse Q01064 gene shares many similar or identical exon boundaries as well as considerable sequence identity with the rat Q07343 and Q08499 genes and the Drosophila dunce DB02527 -specific PDE gene dnc , suggesting that these genes all arose from a common ancestor . Using fluorescence in situ hybridization , we localized the Q01064 gene to the distal tip of mouse Chromosome ( Chr ) 15 . 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 . Glycoprotein IIb/IIIa blockade inhibits platelet-mediated force development and reduces gel elastic modulus . The effects of P08514 /IIIa blockade on clot retraction were studied utilizing an instrument which directly measures force produced by platelets . P08514 /IIIa disruption by calcium chelation , and P08514 /IIIa blockade by peptides and anti- P08514 /IIIa antibodies were investigated . One mM DB00974 suppressed ADP-induced platelet aggregation by 72 % and reduced force developed at 1200 s by 33 % . At 234 microM , the tetrapeptide DB00125 - DB00145 - DB00128 - DB00133 ( RGDS ) suppressed platelet aggregation by 74 % , reduced force at 1200 s by 45 % and reduced gel elastic modulus by 19 % . At 10 microM , the peptide D- DB00125 - DB00145 - DB00128 -L-Try ( D-RGDW ) completely suppressed platelet aggregation , reduced force development by 38 % and reduced gel elastic modulus by 29 % . At 0.133 microM , monoclonal anti- P05106 antibody ( AP-3 ) reduced force development by 74 % and reduced gel modulus by 60 % . Murine antiGPIIb/IIIa antibodies 10E5 and 7E3 markedly suppressed force development . At 0.133 microM , 10E5 reduced force by 89 % and reduced gel modulus by 67 % . At 0.053 microM , 7E3 completely stopped force development and reduced gel modulus by 46 % . Platelet aggregation was blocked by 0.027 microM DB00054 . Selective P08514 blockade by antibodies did not affect force development . None of the agents studied altered fibrin structure as monitored by effects of fibrin mass/length ratios . Suppression of platelet aggregation occurred at inhibitor concentrations substantially lower than those required to suppress force development . Complete suppression of platelet aggregation did not assure inhibition of clot retraction probably due to profound platelet activation by thrombin. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . In vivo imaging visualizes discoid platelet aggregations without endothelium disruption and implicates contribution of inflammatory cytokine and integrin signaling . The mechanism by which thrombotic vessel occlusion occurs independently of plaque development or endothelial cell ( EC ) disruption remains unclear , largely because of an inability to visualize the formation of thrombus , especially at the single-platelet level in real time . Here we demonstrate that rapidly developing thrombi composed of discoid platelets can be induced in the mesenteric capillaries , arterioles , and large-sized arteries of living mice , enabling characterization of the kinetics of thrombosis initiation and the multicellular interrelationships during thrombus development . Platelet aggregation without EC disruption was triggered by reactive oxygen species ( ROS ) photochemically induced by moderate power laser irradiation . The inflammatory cytokines P01375 -α and IL-1 could be key components of the EC response , acting through regulation of P04275 mobilization to the cell surface . Thrombus formation was then initiated by the binding of platelet GPIbα to endothelial P04275 in our model , and this effect was inhibited by the ROS scavenger DB06151 . Actin linker talin-dependent activation of alphaIIb-beta3 integrin or Rac1 in platelets was required for late-phase thrombus stability . Our novel imaging technology illustrates the molecular mechanism underlying inflammation-based thrombus formation by discoid platelets on undisrupted ECs and suggests control of ROS could be a useful therapeutic target for the prevention of thrombotic diseases . 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 . 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 . 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 . 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 . Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins with flow cytometry . Disintegrins are a group of snake venom peptides which inhibit human platelet aggregation by acting as glycoprotein IIb-IIIa ( P08514 -IIIa ) antagonists . They are cysteine-rich , DB00125 - DB00145 - DB00128 ( RGD ) -containing peptides , and bind to P08514 -IIIa complex on platelet membrane with a very high affinity ( Kd , 10(-7)-10(-8) M ) . In this study , we analyzed P08514 -IIIa complex on platelet membrane by flow cytometry using fluorescein isothiocyanate ( FITC ) -conjugated disintegrins as probes . Of these FITC-conjugated disintegrins , FITC-Rhodostomin is the most sensitive probe because Rhodostomin was conjugated with more FITC molecules than Trigramin and Halysin were . The binding fluorescence intensity of FITC-Trigramin ( FITC-Tg ) , FITC-Halysin ( FITC-Hy ) and FITC-Rhodostomin ( FITC-Rn ) was measured in both resting and ADP-activated platelets of diluted human platelet-rich plasma . The binding fluorescence of FITC-disintegrins was abolished by DB00974 and DB00054 , a monoclonal antibody against P08514 -IIIa . ADP markedly increased the fluorescence intensity of FITC-Tg and FITC-Hy bound on platelets especially when lower doses of these probes were used , whereas it had little effect on that of FITC-Rn . Therefore , FITC-Tg and FITC-Hy can be used for the detection of the activated platelets as noted by a higher ratio of fluorescence intensity ( approx. 2-4 ) between ADP-activated and resting platelets as compared with that ( approx. 1-1.3 ) in the case of FITC-Rn as the probe . The platelets from three patients with Glanzmann 's thrombasthenia were probed with FITC-disintegrins. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Using ImageJ for the quantitative analysis of flow-based adhesion assays in real-time under physiologic flow conditions . This article intends to close the gap between the abundance of regular articles focusing on adhesive mechanisms of cells in a flow field and purely technical reports confined to the description of newly developed algorithms , not yet ready to be used by users without programming skills . A simple and robust method is presented for analysing raw videomicroscopic data of flow-based adhesion assays using the freely available public domain software ImageJ . We describe in detail the image processing routines used to rapidly and reliably evaluate the number of adherent and translocating platelets in videomicroscopic recordings . The depicted procedures were exemplified by analysing platelet interaction with immobilized P04275 and fibrinogen in flowing blood under physiological wall shear rates . Neutralizing GPIbalpha function reduced shear-dependent platelet translocation on P04275 and abolished firm platelet adhesion . DB00054 , Tirofiban and DB00063 completely inhibited P08514 /IIIa-dependent stable platelet deposition on fibrinogen . The presented method to analyse videomicroscopic recordings from flow-based adhesion assays offers the advantage of providing a simple and reliable way to quantify flow-based adhesion assays , which is completely based on ImageJ and can easily be applied to study adhesion mechanisms of cells in non-fluorescent modes without the need to deviate from the presented protocol . 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 . A Val193Met mutation in P05106 results in a P08514 /IIIa receptor with a constitutively high affinity for a small ligand . We have identified a patient designated as ( GTa ) with Glanzmann 's Thrombasthenia ( GT ) diagnosed on the basis of a prolonged bleeding time and failure of the patient 's platelets to aggregate . The number of glycoprotein (GP)IIb/IIIa receptors on the platelet surface was 37 % of normal and those receptors displayed a defect in soluble fibrinogen binding . Nevertheless , GTa platelets showed increased adhesion to solid-phase fibrinogen and binding affinity for the RGD-mimetic (3)H-SC52012 , a non-peptide P08514 /IIIa antagonist . Dithiothreitol ( DTT ) and ADP enhanced the affinity for [ (3)H ] -SC52012 in normal platelets , but had little effect in GTa platelets . These findings suggested that GTa platelets were locked in an altered affinity state . Genetic analysis showed that GTa was a compound heterozygote for the P05106 gene . One allele showed a deletion at the 3' end of exon 3 resulting in a premature stop codon . The second P05106 allele had a G to A transition at nucleotide 577 , resulting in a Val193Met substitution . P29320 293T cells transfected with mutant P08514 /IIIaV193M bound [ (3)H ] -SC52012 with a higher affinity than wild-type P08514 /IIIa , and this was not increased by DTT . The mutant receptor distinguishes between platelet adhesion and aggregation , and demonstrates the phenotype that may be expected when platelet aggregation alone is inhibited . 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 . Effect of glycoprotein IIb-IIIa receptor antagonism on platelet membrane glycoproteins after coronary stent placement . Platelet membrane glycoproteins play a crucial role in ischemic complications after coronary stenting . Glycoprotein IIb-IIIa blockade reduces adverse clinical events after angioplasty but is associated with rare but profound thrombocytopenia that might increase hemorrhagic complications . Changes in platelet membrane glycoproteins of patients with angina who underwent coronary stenting and were treated with the P08514 -IIIa antagonist abciximab ( n=20 ) or with heparin ( n=23 ) were studied . GPIb-IIIa receptor blockade and membrane glycoproteins were evaluated with immunological markers in venous blood samples taken before . 10 , 24 , 48 , 72 , and 96 h after initial treatment with either abciximab or heparin . Patients receiving abciximab therapy showed a rapid inhibition of binding of fluorochrome-conjugated mAb CD41 and c7E3 concomitant with a reduction in platelet aggregation which was restored in part in the days after termination of abciximab infusion . Induction of ligand-induced binding sites on P08514 -IIIa was increased in patients receiving abciximab . The expression of ligand-induced binding sites correlated inversely with platelet count . No significant change in platelet membrane markers were found in the heparin group . In vitro studies showed that abciximab induces ligand-induced binding sites on isolated platelets and on nuclear cells bearing recombinant P08514 -IIIa . DB00054 rapidly achieves P08514 -IIIa receptor blockade after coronary stent placement that might be beneficial in high-risk settings to bridge the delayed action of ticlopidine . Significant alterations of platelet membrane glycoproteins during P08514 -IIIa antagonism might contribute to development of acute profound thrombocytopenia . Marinobufagenin regulates permeability and gene expression of brain endothelial cells . Marinobufagenin ( MBG ) is a cardiotonic steroid that increases in the circulation in preeclampsia . Preeclampsia and eclampsia are associated with cerebral edema . Therefore , we examined the effects of MBG on human brain microvascular endothelial cells ( HBMEC ) in vitro . MBG enhanced the permeability of HBMEC monolayers at 1- , 10- , and 100-nM doses , but had no effect at 0.1 nM . Agilent Human Gene Expression microarrays were utilized in these studies . MBG treatment ( 10 nM for 12 h ) downregulated concentrations of the soluble VEGFR transcript sFLT by 59 % but did not alter those of FLTv3 mRNA ( determined by quantitative PCR ) . When treated and control HBMEC transcriptomes were interrogated on microarrays , 1,069 genes appeared to be regulated by MBG . Quantitative RT-PCR confirmed that MBG treatment upregulated Q8TC29 mRNA concentrations by 57 % . Its protein product interacts with calmodulin and calcium channel proteins . MBG treatment downregulated several genes whose protein products are involved in cell adhesion ( P08514 , Q9BQL6 , Q9Y5I7 , and Q6UWW9 ) and cell signaling ( Q14957 , P32418 , and P03372 ) . The level of downregulation ranged from 22 to 66 % . Altogether , MBG actively enhanced the permeability of HBMEC monolayers while downregulating genes involved in adhesion . MBG treatment had variable effects on Q8TC29 , Q14957 , and P32418 genes , all associated with calcium transport . These studies provide the basis for future investigations of MBG actions in normal physiology and disease . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . [ 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 . 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 . 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 ) [ 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 . Purification and characterization of platelet aggregation inhibitors from snake venoms . Proteins that inhibit glycoprotein ( GP ) IIb/IIIa mediated platelet aggregation have been purified from the venom of two snake species . A small platelet aggregation inhibitor ( p1.AI ) , multisquamatin ( Mr = 5,700 ) , was purified from Echis multisquamatus venom by hydrophobic interaction HPLC and two steps on C18 reverse phase HPLC . A larger p1.AI , contortrostatin ( Mr = 15,000 ) , was purified by a similar HPLC procedure from the venom of Agkistrodon contortrix contortrix . Both p1.AIs inhibit ADP-induced human , canine and rabbit platelet aggregation using platelet rich plasma ( PRP ) . Multisquamatin has an IC50 of 97 nM , 281 nM and 333 nM for human , canine and rabbit PRP , respectively . Contortrostatin has an IC50 of 49 nM , 120 nM and 1,150 nM for human , canine and rabbit PRP , respectively . In a competitive binding assay using 125I- DB00054 ( a monoclonal antibody to P08514 /IIIa that inhibits platelet aggregation ) both contortrostatin and multisquamatin demonstrated P08514 /IIIa specific binding to human and canine platelets . The IC50 for contortrostatin displacement of 7E3 binding to human and canine P08514 -/IIIa is 27 nM and 16 nM , respectively and for multisquamatin it is 3 nM and 63 nM , respectively . Our results indicate that both p1.AIs inhibit platelet aggregation by binding with high affinity to P08514 /IIIa . 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 . 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 . 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 . 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 . 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 . 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 . [ New immunological weapons for medicine in the 21st Century : biological therapy based on the use of the latest generation monoclonal antibodies ] . The fusion of a murine B cell and a myeloma cell generates a hybridoma that produces monoclonal antibody ( mAb ) . These murine mAb induce the HAMA ( human anti-mouse antibodies ) response . Murine mAb have been modified by genetic engineering , producing molecules with a higher proportion of human protein . At present , chimeric , humanized and fully human mAb are available . mAb block interactions between target molecules and their ligands or trigger the lyses of mAb-coated tumor cells . Numerous mAb have been developed using the recombinant DNA technology and several are available in the market . DB00072 , against P04626 /neu , is useful in breast cancer ; rituximab , against P11836 in B lymphocytes is useful in lymphoma ; alemtuzumah , against P31358 is used in lymphoma and leukemia ; daclizumab and basiliximab block the P60568 receptor interaction and reduce acute rejection in kidney transplantation ; abciximab , an antagonist of P08514 /IIIa platelet receptor , is used in patients undergoing acute coronary syndromes . In autoimmunity diseases , blocking tumor necrosis factor by infliximab and adalimumab has demonstrated excellent results . Thus , infliximab is useful in the treatment of rheumatoid arthritis ( RA ) , Crohn 's disease and ulcerative colitis while adalimumab is the first fully human mAb available for RA . DB00065 and adalimumab reduce signs and symptoms in RA and they also interfere with progression of joint damage . Finally , the direct benefits of antagonist treatment can occur at the expense of a major adverse effect in some other biological function . 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 . 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 . A new death domain associated with gestational trophoblastic diseases induces apoptosis in distinct cell types . Gestational trophoblastic diseases , like the complete hydatidiform mole ( P24386 ) , are a group of human interrelated neoplasms whose etiology and progression is poorly understood at the molecular level . We have previously reported the cloning and expression of a new tumor necrosis factor receptor ( P01375 -R ) related transcript , named CHMS-1 that encodes a potential death domain . Here we show that ectopic expression of the putative CHMS-1 death domain specifically induced apoptosis in a dose-dependent manner , in trophoblastic ( JEG-3 ) and non-trophoblastic ( COS-7 ) cells . We also investigated the expression of apoptosis-related molecules such as Bcl-2 and p53 and demonstrated that Bcl-2 is repressed in P24386 while p53 is overexpressed in P24386 compared with persistent gestational trophoblastic tumors . Altogether , these data indicate that the CHMS-1 death domain is able to trigger apoptosis , thus suggesting that this new entity might be an important inducer of molar regression mechanisms in women . 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 .

MH_train_1127

MH_train_1127

MH_train_1127

interacts_with DB01045?

multiple_choice

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 % . 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 . 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 . TGFbeta1 , TNFalpha , and insulin signaling crosstalk in regulation of the rat cholesterol 7alpha-hydroxylase gene expression . The TGFbeta1/Smad pathway plays a critical role in cholestasis and liver fibrosis . Previous studies show that TGFbeta1 , TNFalpha , and insulin inhibit cholesterol 7alpha-hydroxylase ( P22680 ) gene transcription and bile acid synthesis in human hepatocytes . In this study , we investigated insulin , TGFbeta1 , and TNFalpha regulation of rat Cyp7a1 gene transcription . In contrast to inhibition of human P22680 gene transcription , TGFbeta1 stimulates rat Cyp7a1 reporter activity . P84022 , FoxO1 , and HNF4alpha synergistically stimulated rat Cyp7a1 gene transcription . Mutations of the P84022 , FoxO1 , or HNF4alpha binding site attenuated the rat Cyp7a1 promoter activity . Furthermore , TNFalpha and cJun attenuated TGFbeta1 stimulation of rat Cyp7a1 . P01308 or adenovirus-mediated expression of constitutively active P31749 inhibited FoxO1 and P84022 synergy . In streptozotocin-induced diabetic rats , Cyp7a1 mRNA expression levels were induced and insulin attenuated P22680 mRNA levels . Chromatin immunoprecipitation assay showed that FoxO1 binding to Cyp7a1 chromatin was increased in diabetic rat livers and insulin reduced FoxO1 binding . These results suggest a mechanistic basis for induction of Cyp7a1 activity and bile acid synthesis in cholestatic rats and in diabetic rats . The crosstalk of insulin , TGFbeta and TNFalpha signaling pathways may regulate bile acid synthesis and lipid homeostasis in diabetes , fatty liver disease , and liver fibrosis . Targeting mitochondrial 18 kDa translocator protein ( TSPO ) regulates macrophage cholesterol efflux and lipid phenotype . The aim of the present study was to establish mitochondrial cholesterol trafficking 18 kDa translocator protein ( TSPO ) as a potential therapeutic target , capable of increasing macrophage cholesterol efflux to (apo)lipoprotein acceptors . Expression and activity of TSPO in human ( THP-1 ) macrophages were manipulated genetically and by the use of selective TSPO ligands . Cellular responses were analysed by quantitative PCR ( Q-PCR ) , immunoblotting and radiolabelling , including [3H]cholesterol efflux to (apo)lipoprotein A-I ( apoA-I ) , high-density lipoprotein ( HDL ) and human serum . Induction of macrophage cholesterol deposition by acetylated low-density lipoprotein ( AcLDL ) increased expression of TSPO mRNA and protein , reflecting findings in human carotid atherosclerosis . Transient overexpression of TSPO enhanced efflux ( E % ) of [3H]cholesterol to apoA-I , HDL and human serum compared with empty vector ( EV ) controls , whereas gene knockdown of TSPO achieved the converse . Ligation of TSPO ( using PK11195 , FGIN-1-27 and flunitrazepam ) triggered increases in [3H]cholesterol efflux , an effect that was amplified in TSPO-overexpressing macrophages . Overexpression of TSPO induced the expression of genes [ Q07869 ( peroxisome-proliferator-activated receptor α ) , Q13133 ( nuclear receptor 1H3/liver X receptor α ) , O95477 ( DB00171 -binding cassette A1 ) , Q9H172 ( DB00171 -binding cassette G4 ) and P02649 ( apolipoprotein E ) ] and proteins ( O95477 and PPARα ) involved in cholesterol efflux , reduced macrophage neutral lipid mass and lipogenesis and limited cholesterol esterification following exposure to AcLDL . Thus , targeting TSPO reduces macrophage lipid content and prevents macrophage foam cell formation , via enhanced cholesterol efflux to (apo)lipoprotein acceptors . 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 . Down-regulation of cholesterol 7alpha-hydroxylase ( P22680 ) gene expression by bile acids in primary rat hepatocytes is mediated by the c-Jun N-terminal kinase pathway . DB04540 7alpha-hydroxylase ( P22680 ) , the rate-limiting enzyme in the neutral pathway of bile acid biosynthesis , is feedback-inhibited at the transcriptional level by hydrophobic bile acids . Recent studies show that bile acids are physiological ligands for farnesoid X receptor ( Q96RI1 ) . Activated Q96RI1 indirectly represses P22680 transcription through induction of small heterodimer protein ( Q15466 -1 ) . In this study , we provide evidence that bile acids rapidly down-regulate P22680 transcription via activation of the JNK/c-Jun pathway . Furthermore , we demonstrate that Q15466 -1 is also a direct target of activated c-Jun . In primary rat hepatocyte cultures , taurocholate ( TCA ) strongly activated JNK in a time- and concentration-dependent manner . P01375 -alpha , a potent activator of JNK , also rapidly activated JNK and down-regulated P22680 mRNA levels . Overexpression of dominant-negative P45983 or a transactivating domain mutant of c-Jun significantly blocked the ability of TCA to down-regulate P22680 mRNA . In contrast , overexpression of wild-type c-Jun ( c-Jun(wt) ) enhanced the repression of P22680 by TCA . Moreover , overexpression of c-Jun(wt) resulted in increased Q15466 -1 promoter activity . Mutation of a putative AP-1 ( c-Jun ) element suppressed c-Jun-mediated activation of the Q15466 -1 promoter construct . These results indicate that the bile acid-activated JNK pathway plays a pivotal role in regulating P22680 levels in primary rat hepatocytes . 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 . 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 . 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 . DB00624 response of hepatic gene expression in female mice having acquired testosterone-unresponsive immunity to Plasmodium chabaudi malaria . Blood-stage malaria of Plasmodium chabaudi is characterized by its responsiveness to testosterone ( T ) : T suppresses development of protective immunity , whereas once acquired immunity is T-unresponsive . Here , we have analyzed the liver , a T target and lymphoid organ with anti-malaria activity , for its T-responsiveness of gene expression in immune mice . Using Affymetrix microarray technology , in combination with quantitative RT-PCR , we have identified ( i ) T-unresponsive expression of newly acquired mRNAs encoding diverse sequences of IgG- and IgM-antibodies , ( ii ) 24 genes whose expression has become T-unresponsive including those encoding the T(H)2 response promoting Q96KQ7 and the erythrocyte membrane protein band 7.2 P27105 , ( iii ) T-unresponsive expression of mRNAs for the cytokines IL-1β , P05231 , TNFα , and IFNγ , as well as P35228 , which are even not inducible by infection , and ( iv ) 35 genes retaining their T-responsiveness , which include those encoding the infection-inducible acute phase proteins P0DJI8 , P0DJI9 , and P19652 as well as those of liver metabolism which encode the T-downregulated female-prevalent enzymes CYP2B9 , CYP2B13 , CYP3A41 , P22680 , and SULT2A2 and the T-upregulated male-prevalent enzymes CYP2D9 , O75881 , UGT2B1 , P26439 , HSD3B5 , respectively . The mRNA of the latter T-metabolizing enzyme is even 5-fold increased by T , suggesting a decrease in the effective T concentrations in the liver of immune mice . Collectively , our data suggest that the liver , which has acquired a selective T-unresponsiveness of gene expression , contributes to the acquired T-unresponsive , antibody-mediated protective immunity to blood-stage malaria of P. chabaudi . 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 . 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 . [ Clinical and genetic characteristics of long-livers in Moscow region ] . In Moscow region long-livers we have studied distribution of P06858 , P11597 , P02649 , F2 , P12259 , P08709 , F13 , P02675 , P17301 , P05106 , P05121 , P42898 , Q9UBK8 , HLA- Q8IUH3 , HLA-DQA1 , P01920 genes polymorphisms , associated with predisposition to age pathology . Long-livers are characterized by favorable course of cardiovascular diseases accompanied by certain genetic factors . We have established that genotype H-H- of P06858 , allele epsilon2 of P02649 , genotype CC of P42898 ( 677C > T ) , genotype TC of P05106 , genotype GA of P02675 , HLA- Q8IUH3 *11 positively correlate with longevity . Suppression of dendritic cell maturation by Trichinella spiralis excretory/secretory products . Evidence from experimental studies indicates that during chronic infections with certain helminth species a regulatory network is induced that can down-modulate not only parasite-induced inflammation but also reduce other immunopathologies such as allergies and autoimmune diseases . The mechanisms however , and the molecules involved in this immunomodulation are unknown . Here , we focus on the effect of Trichinella spiralis excretory/secretory antigens ( TspES ) on the innate immune response by studying the effect of TspES on DC maturation in vitro . Bone marrow-derived DC from BALB/c mice were incubated with TspES either alone or in combination with LPS derived from two different bacteria . As indicators of DC maturation , the cytokine production ( IL-1alpha , P05231 , P22301 , IL-12p70 and P01375 ) and the expression of various surface molecules ( MHC-II , P25942 , P33681 and P42081 ) were measured . Results indicate that while TspES alone did not change the expression of the different surface molecules or the cytokine production , it completely inhibited DC maturation induced by Escherichia coli LPS ( E. coli LPS ) . In contrast , DC maturation induced by LPS from another bacterium , Neisseria meningitidis , was not affected by TspES . These results were confirmed using O00206 / Q9Y6Y9 / P08571 transfected P29320 293 cells . In conclusion , T. spiralis ES antigens lead to suppression of DC maturation but this effect depends on the type of LPS used to activate these cells . [ Serotonin transporter gene and stress reactivity in unipolar depression. Role of the Q9Y251 system as endophenotype of the P31645 gene ] . BACKGROUND : A length polymorphism in the promoter region of the serotonin transporter gene ( 5-HTTLPR ) is associated with both depression and hypothalamic-pituitary-adrenal ( Q9Y251 ) system activity . A dysregulation of the Q9Y251 system is considered to be a candidate endophenotype of depression . The objective of the present study was an investigation of a possible gene-endophenotype-interaction between 5-HTTLPR and Q9Y251 system activity in a sample of inpatients with major depression . MATERIALS AND METHODS : A total of 237 inpatients with major depression were genotyped for 5-HTTLPR and participated in a combined dexamethasone-corticotropin-releasing hormone test ( DB00514 - P06850 test ) as well as using the Hamilton score ( Hamilton rating scale for depression ) to determine the severity of the psychopathology . RESULTS : Patients with the ss-genotype showed a significantly higher Q9Y251 -system activity in comparison to patients with the lI-genotype , but no association between 5-HTTLPR and the severity of psychopathology could be detected . CONCLUSIONS : The results of the current study demonstrate an influence of 5-HTTLPR on dysregulation of the Q9Y251 system in patients with major depression and support the hypothesis that 5-HTTLPR- and Q9Y251 -system-interaction constitutes an important component in the pathogenesis of depression . 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 . Nuclear hormone receptors and cholesterol trafficking : the orphans find a new home . There are many homeostatic mechanisms that contribute to the regulation of cellular and serum cholesterol levels in humans . Much of our understanding of this regulation arose from studies of the cellular pathways controlling cholesterol synthesis and the uptake and degradation of low-density lipoproteins . The physiology governing cholesterol disposition in whole animals , including the molecular mechanisms responsible for dietary uptake of cholesterol and its subsequent catabolism to bile acids , are only now being uncovered . This review summarizes recent studies that have used modern genetic techniques , as well as cell biological methods , to begin to elucidate the pathways responsible for cholesterol trafficking in vivo . This work has led to the realization that networks of nuclear hormone receptors , including the LXR , Q96RI1 , Q07869 , and RXR proteins , play critical roles in lipid metabolism by virtue of their transcriptional regulation of the genes that control sterol metabolic pathways . Some of the major downstream targets of these regulatory networks involve members of the ABC transporter family , including O95477 , P45844 , Q9H222 , Q9H221 , P21439 /Mdr2 , and SPGP/ O95342 . These transporters contribute to the movement of sterols and biliary lipids across cellular membranes via mechanisms that have yet to be elucidated . The potential for manipulating these cholesterol trafficking pathways via drugs targeted to the nuclear hormone receptors has generated great interest in their biology and will undoubtedly lead to new therapeutic approaches to human disorders in the coming years . Clinical significance of arginase after liver transplantation . Liver graft function after transplantation is dependent on ischemia-reperfusion injury , toxicity of drugs ( immunosuppression , antibiotics and other ) and transplant rejection . Although routinely monitored with enzymatic tests ( Q9NRA2 , ALT , P19440 , ALP ) , bilirubin and coagulation parameters , differentiation between these pathologies is hardly possible without liver biopsy . Arginase ( 3.5.3.1 ) mostly exists in the liver and in trace amounts in extra-hepatic tissue . Thus , we hypothesized that activity of arginase could be a more specific test of liver function . Sera of 32 liver transplant recipients were tested for Q9NRA2 , ALT , P01008 , bilirubin and arginase . Samples were obtained daily in first 2 weeks after LTx and weekly afterwards . Correlation of arginase activity with other liver function markers was calculated . Serum arginase peaked at day 1 post LTx ( mean 64,6+/-91 IU/L ) , and decreased more rapidly than other tests if good liver function was observed . The values showed strong and significant correlation with Q9NRA2 and ALT activities ( Pearsons R 0,65 and 0,47 respectively ) . We conclude that activity of arginase in the serum is an exact test of liver function . 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 . 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 . 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 . 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 . 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 . 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 . The genomic clone P08908 which resembles a beta-adrenergic receptor sequence encodes the P08908 receptor . The recent cloning of the complementary DNAs and/or genes for several receptors linked to guanine nucleotide regulatory proteins including the adrenergic receptors ( alpha 1 , alpha 2A , alpha 2B , beta 1 , beta 2 ) , several subtypes of the muscarinic cholinergic receptors , and the visual ' receptor ' rhodopsin has revealed considerable similarity in the primary structure of these proteins . In addition , all of these proteins contain seven putative transmembrane alpha-helices . We have previously described a genomic clone , P08908 , isolated by cross-hybridization at reduced stringency with a full length beta 2-adrenergic receptor probe . This clone contains an intronless gene which , because of its striking sequence resemblance to the adrenergic receptors , is presumed to encode a G-protein-coupled receptor . Previous attempts to identify this putative receptor by expression studies have failed . We now report that the protein product of the genomic clone , Q96NT5 , transiently expressed in monkey kidney cells has all the typical ligand-binding characteristics of the 5-hydroxytryptamine ( P08908 ) receptor . 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 . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . 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 . 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 . Chemotactic recruitment of adult neural progenitor cells into multifunctional hydrogels providing sustained SDF-1α release and compatible structural support . Without chemotactic cues and structural support , cavitary brain lesions typically fail to recruit endogenous neural progenitor cells ( NPCs ) . Toward resolving this , we engineered multifunctional biomaterials comprising injectable gelatin-hydroxyphenylpropionic acid ( Gtn- Q9Y251 ) hydrogels and dextran sulfate/chitosan polyelectrolyte complex nanoparticles ( PCNs ) that delivered stromal cell-derived factor-1α ( SDF-1α ) . Over 7 d of interface with in vitro tissue simulant containing adult rat hippocampal NPCs ( aNPCs ) and their neuronal progeny , Gtn- Q9Y251 /SDF-1α- Q15149 hydrogels promoted chemotactic recruitment to enhance infiltration of aNPCs by 3- to 45-fold relative to hydrogels that lacked SDF-1α or vehicles to sustain SDF-1α release . When cross-linked with 0.85-0.95 mM HO , Gtn- Q9Y251 /SDF-1α- Q15149 hydrogels provided optimally permissive structural support for migration of aNPCs . Specific matrix metalloproteinase ( MMP ) inhibitors revealed that 42 , 30 , and 55 % of cell migration into Gtn- Q9Y251 /SDF-1α- Q15149 hydrogels involved P08253 , 3 , and 9 , respectively , demonstrating the hydrogels to be compatible toward homing endogenous NPCs , given their expression of similar MMPs . Interestingly , PCNs utilized P09038 found in situ to induce chemokinesis , potentiate SDF-1α chemotactic recruitment , and increase proliferation of recruited cells , which collectively orchestrated a higher number of migrated aNPCs . Overall , Gtn- Q9Y251 /SDF-1α- Q15149 hydrogels prove to be promising biomaterials for injection into cavitary brain lesions to recruit endogenous NPCs and enhance neural tissue repair/regeneration . 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 . P02647 inhibits P25942 proinflammatory signaling via DB00171 -binding cassette transporter A1-mediated modulation of lipid raft in macrophages . AIM : P02647 ( apoA-I ) , the major component of high-density lipoprotein ( HDL ) , has been recently found to suppress inflammation . This study was to investigate the effects and potential mechanisms of apoA-I on the P25942 / P29965 ( P29965 ) proinflammatory signaling pathway . METHODS : Human THP-1 macrophage-derived foam cells were treated with sCD40L alone or in the presence of apoA-I . Secretion of proinflammatory cytokines was performed by enzyme-linked immunosorbent assay(ELISA) . The proteins and mRNA expression were examined by western-blot and real-time PCR analysis , respectly . DB04540 efflux was assessed by liquid scintillation counting . DB04540 depletion of macrophages was performed with methylated β-cyclodextrin . RESULTS : P02647 inhibits the inflammatory response stimulated by soluble P29965 ( sCD40L ) in macrophages . In addition , apoA-I inhibited the sCD40L-stimulated activation of nuclear factor-kB ( NF-kB ) . The apoA-I-induced NF-kB deactivation was related to the decreased recruitment of tumor necrosis factor receptor-associated factor 6 ( TRAF-6 ) , a crucial adapter protein for P25942 in macrophages , to lipid rafts after being treated by sCD40L . When interfering the expression of DB00171 -binding cassette transporter A1 ( O95477 ) , a major cholesterol transporter for apoA-I in macrophages , it could significantly diminish the effect of apoA-I on the sCD40L-stimulated inflammatory response . CONCLUSION : P02647 suppresses P25942 proinflammatory signaling in macrophages by preventing TRAF-6 translocation to lipid rafts through O95477 -dependent regulation of free cholesterol ( FC ) efflux , which may present a novel mechanism of apoA-I-mediated inflammation inhibition in macrophages . 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 . 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 . 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 . 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 . 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 . 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 . Two different P01579 nonresponsive variants derived from the B-cell lymphoma 70Z/3 . The kappa immunoglobulin ( Igk ) light chain locus is transcriptionally silent in the mouse B-cell lymphoma 70Z/3 . However , exposure to lipopolysaccharide ( LPS ) or interferon-gamma ( IFN ) causes a marked increase in Igk transcription . By immunoselection , we isolated two variants that are nonresponsive to IFN . One variant , AT7.2 , has retained its response to LPS ( IFN-LPS+ ) , whereas the other , P01008 .3 , is also nonresponsive to LPS ( IFN-LPS- ) . Stable transfection of an intact Igk gene does not rescue the phenotype of either variant . Both variants have intact Igk genes and neither is deficient in the binding or uptake of IFN . Nuclear extracts from LPS-treated wild-type 70Z/3 cells show strong increases in three transcription factors : P09086 , NF-kappa B , and kBF-A . Remarkably , when the IFN-LPS- variant is treated with LPS , all three transcription factors are still observed in the nuclear extracts . Treatment of wild-type cells with either LPS or IFN also causes a decrease in nuclear complexes that bind to two other regions of the Igk intron enhancer , the octenh and the E kappa MHCIC regions . Both of these changes are also observed after LPS or IFN treatment of the IFN-LPS- variant . Thus , this variant transduces the IFN and LPS signals at least into the nuclear compartment , but still fails to activate Igk transcription . In contrast , the IFN-LPS+ variant decreases neither the octenh nor the E kappa MHCIC binding complexes in response to IFN . This variant may be defective in transducing the IFN signal to the nucleus . These variants will be useful in studying the activation of Igk transcription and the IFN signaling pathway in B cells . Functional characterization of a novel serotonin receptor ( 5-HTap2 ) expressed in the CNS of Aplysia californica . Serotonin has been shown to be a neuromodulator in the Aplysia californica CNS . The diversity of serotonin actions is due to the existence of several different receptor subtypes . In this study we report the cloning of a full-length cDNA , coding for a novel serotonin receptor ( 5-HTap2 ) . The receptor protein bears the characteristics of G protein-coupled receptors . It shares 68 % and 34 % of its amino acid sequence identity with the 5-HTlym receptor from Lymnaea stagnalis and the mammalian P08908 receptor , respectively . When transfected in P29320 293 cells , 5-HTap2 was negatively coupled to adenylate cyclase . Ligand binding analysis indicated that the order of potencies of various drugs for the inhibition of [3H]LSD binding was : methiothepin > metergoline > 5-CT > PAPP > 5-HT > ketanserin > NAN-190 > 8-OH-DPAT > clozapine . RT-PCR amplification of RNA isolated from different tissues indicated that this receptor is expressed in the CNS and in bag cells . The expression of 5-HTap2 restricted to the CNS suggests an important role for this receptor in the modulation of neuronal functions in Aplysia . Moreover , the high expression of 5-HTap2 in the bag cells , associated with its pharmacological profile , suggests that this receptor may be implicated in modulating the afterdischarge during the egg-laying behavior . 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 . 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 . 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 . Progressive familial intrahepatic cholestasis . Progressive familial intrahepatic cholestasis ( O43520 ) is a group of rare disorders which are caused by defect in bile secretion and present with intrahepatic cholestasis , usually in infancy and childhood . These are autosomal recessive in inheritance . The estimated incidence is about 1 per 50,000 to 1 per 100,000 births , although exact prevalence is not known . These diseases affect both the genders equally and have been reported from all geographical areas . Based on clinical presentation , laboratory findings , liver histology and genetic defect , these are broadly divided into three types- O43520 type 1 , O43520 type 2 and O43520 type 3 . The defect is in O43520 gene encoding the O43520 protein , ABCB 11 gene encoding O95342 protein and P21439 gene encoding P21439 protein in PFIC1 , 2 and 3 respectively . The basic defect is impaired bile salt secretion in PFIC1/2 whereas in PFIC3 , it is reduced biliary phospholipid secretion . The main clinical presentation is in the form of cholestatic jaundice and pruritus . Serum gamma glutamyl transpeptidase ( P19440 ) is normal in patients with PFIC1/2 while it is raised in patients with PFIC3 . Treatment includes nutritional support ( adequate calories , supplementation of fat soluble vitamins and medium chain triglycerides ) and use of medications to relieve pruritus as initial therapy followed by biliary diversion procedures in selected patients . Ultimately liver transplantation is needed in most patients as they develop progressive liver fibrosis , cirrhosis and end stage liver disease . Due to the high risk of developing liver tumors in PFIC2 patients , monitoring is recommended from infancy . Mutation targeted pharmacotherapy , gene therapy and hepatocyte transplantation are being explored as future therapeutic options . (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 . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . 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 . P10275 -mediated regulation of the anti-atherogenic enzyme Q02318 involves the JNK/c-jun pathway . Q02318 , an enzyme with several important roles in cholesterol homeostasis and vitamin D₃ metabolism , has been ascribed anti-atherogenic properties . This study addresses an important problem regarding how this enzyme , involved in cholesterol metabolism in the liver and peripheral tissues , is regulated . Our results identify the human Q02318 gene as a new target for the JNK/c-jun pathway . Initial experiments showed that an inhibitor of c-Jun N-terminal kinase ( JNK ) downregulated basal Q02318 promoter activity whereas overexpression of JNK slightly enhanced promoter activity . P10275 ( AR ) -mediated upregulation of mRNA levels and endogenous enzyme activity was recently reported . In the present study , the AR antagonist nilutamide blocked the androgen induction of Q02318 . The present data revealed that inhibition of the JNK/c-jun pathway abolishes the AR-mediated effect on Q02318 transcription and enzyme activity , whereas overexpression of JNK markedly increased androgenic upregulation of Q02318 . In conclusion , the current results indicate involvement of the JNK/c-jun pathway in AR-mediated upregulation of human Q02318 . The link to JNK signaling is interesting since inflammatory processes may upregulate Q02318 to clear cholesterol from peripheral tissues . Single nucleotide polymorphisms in P11597 , Q96NT5 , P41440 , P16671 , Q9HAY6 , Q6Q788 , and O95477 are significant predictors of plasma HDL in healthy adults . BACKGROUND : In a marker-trait association study we estimated the statistical significance of 65 single nucleotide polymorphisms ( SNP ) in 23 candidate genes on HDL levels of two independent Caucasian populations . Each population consisted of men and women and their HDL levels were adjusted for gender and body weight . We used a linear regression model . Selected genes corresponded to folate metabolism , vitamins B-12 , A , and E , and cholesterol pathways or lipid metabolism . METHODS : Extracted DNA from both the Sacramento and Beltsville populations was analyzed using an allele discrimination assay with a MALDI-TOF mass spectrometry platform . The adjusted phenotype , y , was HDL levels adjusted for gender and body weight only statistical analyses were performed using the genotype association and regression modules from the SNP Variation Suite v7 . RESULTS : Statistically significant SNP ( where P values were adjusted for false discovery rate ) included : P11597 ( rs7499892 and rs5882 ) ; Q96NT5 ( rs37514694 ; rs739439 ) ; P41440 ( rs3788199 ) ; P16671 ( rs3211956 ) ; Q9HAY6 ( rs6564851 ) , Q6Q788 ( rs662799 ) , and O95477 ( rs4149267 ) . Many prior association trends of the SNP with HDL were replicated in our cross-validation study . Significantly , the association of SNP in folate transporters ( Q96NT5 rs37514694 and rs739439 ; P41440 rs3788199 ) with HDL was identified in our study . CONCLUSIONS : Given recent literature on the role of niacin in the biogenesis of HDL , focus on status and metabolism of B-vitamins and metabolites of eccentric cleavage of β-carotene with lipid metabolism is exciting for future study . An intricate network of conserved DNA upstream motifs and associated transcription factors regulate the expression of uromodulin gene . PURPOSE : P07911 is a kidney specific glycoprotein whose expression can modulate kidney homeostasis . However , the set of sequence specific transcription factors that regulate the uromodulin gene P07911 and their upstream binding locations are not well characterized . We built a high resolution map of its transcriptional regulation . MATERIALS AND METHODS : We applied in silico phylogenetic footprinting on the upstream regulatory regions of a diverse set of human P07911 orthologs to identify conserved binding motifs and corresponding position specific weight matrices . We further analyzed the predicted binding motifs by motif comparison , which identified transcription factors likely to bind these discovered motifs . Predicted transcription factors were then integrated with experimentally known protein-protein interactions available from public databases and tissue specific expression resources to delineate important regulators controlling P07911 expression . RESULTS : Analysis allowed the identification of a reliable set of binding motifs in the upstream regulatory regions of P07911 to build a high confidence compendium of transcription factors that could bind these motifs , such as P23771 , P35680 , SP1 , P84022 , Q13950 and O43474 . ENCODE deoxyribonuclease I hypersensitivity sites in the P07911 upstream region of the mouse kidney confirmed that some of these binding motifs were open to binding by predicted transcription factors . The transcription factor-transcription factor network revealed several highly connected transcription factors , such as SP1 , Q02447 , P04637 , P14859 , P10826 , P10276 and P19793 , as well as the likely protein complexes formed between them . Expression levels of these transcription factors in the kidney suggest their central role in controlling P07911 expression . CONCLUSIONS : Our findings will form a map for understanding the regulation of uromodulin expression in health and disease . 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 . 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 . 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 . 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 . 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 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 . 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 . 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 . Involvement of transcription factor P24468 in human trophoblast differentiation . BACKGROUND : During the in vitro differentiation of human villous cytotrophoblast ( Q01459 ) cells to a syncytiotrophoblast ( STB ) phenotype , mRNA levels for the nuclear hormone receptor P24468 ( P24468 , COUP-TFII ) increase rapidly , reaching a peak at day 1 of differentiation that is 8.8-fold greater than that in undifferentiated Q01459 cells . To examine whether P24468 is involved in the regulation of villous Q01459 cell differentiation , studies were performed to determine whether P24468 regulates the expression of P05549 ( AP-2alpha ) , a transcription factor that is critical for the terminal differentiation of these cells to a STB phenotype . METHODOLOGY/PRIMARY FINDINGS : Overexpression of P24468 in primary cultures of human Q01459 cells and JEG-3 human choriocarcinoma cells induced dose-dependent increases in P05549 promoter activity . Conversely , siRNA mediated silencing of the P24468 gene in villous Q01459 undergoing spontaneous differentiation blocked the induction of the mRNAs for P05549 and several STB cell specific marker genes , including human placental lactogen ( hPL ) , pregnancy specific glycoprotein 1 ( P11464 ) and corticotropin releasing hormone ( P06850 ) by 51-59 % . The induction of P05549 promoter activity by P24468 was potentiated by the nuclear hormone receptors retinoic acid receptor alpha ( P10276 ) and retinoid X receptor alpha ( P19793 ) . CONCLUSIONS/SIGNIFICANCE : Taken together , these results strongly suggest that P24468 is involved in villous Q01459 cell differentiation and that P24468 acts , at least in part , by directly activating P05549 gene expression and by potentiating the transactivation of P05549 by P10276 and P19793 . 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 . 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 .

MH_train_1128

MH_train_1128

MH_train_1128

interacts_with DB00243?

multiple_choice

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 . 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 . Growth rates of hprt and tk mutant CHO cell lines . The growth rates of 31 X-ray-induced hypoxanthine phosphoribosyl transferase ( hprt ) deficient mutants of CHO- P04264 cells were measured . Mutants had been classified as ( 1 ) full-deletion , ( 2 ) partial deletion or rearrangement , or ( 3 ) unchanged by Southern blot analyses . No relationship between growth rate and deletion type was observed . Even where all hprt-specific bands were missing , proliferation rates in culture were normal . Additionally , in CHO- P01008 -2 cells , which are heterozygous at the tk locus , no difference in growth rates between a spontaneous hprt mutant and its parent was observed , although double hprt-tk-/- mutants grew more slowly . 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 . 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 . 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 . A novel strategy for the generation of angiostatic kringle regions from a precursor derived from plasminogen . In this study we have explored the feasibility of generating angiostatin by incorporating an endoproteolytic furin cleavage site into plasminogen to allow conversion of the precursor molecule into an angiostatic active P04264 -3 fragment . We show that secretable angiostatin can be successfully generated from cells infected with adenovirus carrying the furin-mutated plasminogen ( AdmuthPlgK3 ) . Supernatant from cells transduced with AdmuthPlagK3 inhibits tube formation and proliferation and migration of human umbilical vein endothelial cells with an efficiency similar to that of supernatant from cells infected with adenovirus expressing kringle 1-3 of plasminogen ( AdK1-3 ) . Administration of AdmuthPlgK3 and AdK1-3 in mice results in significantly decreased endothelial cell infiltration in P15692 -embedded Matrigel plugs . Treatment with AdmuthPlgK3 and AdK1-3 exerts strong antitumoral effect in models of hepatocellular carcinoma and Lewis lung cancer . This antitumor effect was associated with decreased microvessel density in the tumors . Taken together , our data demonstrate that angiostatin endowed with strong antiangiogenic and antitumor effects can be released from a furin-mutated plasminogen acting as a precursor . This strategy may have potential to develop angiostatic anti-cancer therapies . Kinetics of the initial steps of G protein-coupled receptor-mediated cellular signaling revealed by single-molecule imaging . We report on an in vivo single-molecule study of the signaling kinetics of G protein-coupled receptors ( GPCR ) performed using the neurokinin 1 receptor ( P25103 ) as a representative member . The P25103 signaling cascade is triggered by the specific binding of a fluorescently labeled agonist , DB05875 ( SP ) . The diffusion of single receptor-ligand complexes in plasma membrane of living P29320 293 cells is imaged using fast single-molecule wide-field fluorescence microscopy at 100 ms time resolution . Diffusion trajectories are obtained which show intra- and intertrace heterogeneity in the diffusion mode . To investigate universal patterns in the diffusion trajectories we take the ligand-binding event as the common starting point . This synchronization allows us to observe changes in the character of the ligand-receptor-complex diffusion . Specifically , we find that the diffusion of ligand-receptor complexes is slowed down significantly and becomes more constrained as a function of time during the first 1000 ms . The decelerated and more constrained diffusion is attributed to an increasing interaction of the GPCR with cellular structures after the ligand-receptor complex is formed . [ 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 . Encapsulation of viral vectors for gene therapy applications . In gene therapy , a number of viruses are currently being used as vectors to provide transient expression of therapeutic proteins . A drawback of using free virus is that it gives a potent immune response , which reduces gene transfer and limits re-administration . An alternative delivery system is to encapsulate the virus in poly(lactide-co-glycolide) ( P00747 ) microspheres prior to administration . A recombinant adenovirus ( Ad ) expressing green fluorescent protein ( GFP ) was used to test the transduction efficiency of Ad encapsulated in microspheres on target cells . The number of infected cells that expressed GFP was measured by flow cytometry . It was demonstrated that encapsulated viral vectors could successfully transduce target cells with encapsulation efficiencies up to 23 % and that the level of transduction could be controlled by varying both the quantity of microspheres and the amount of Ad in the microspheres . High transduction efficiencies and its recognized biocompatibility make P00747 -encapsulated Ad an attractive alternative to the use of free virus in gene therapy applications . The infectivity of Ad was found to be significantly influenced by the processing conditions and changes in environmental factors . Free Ad and encapsulated Ad were able to infect both E1 complimenting cells ( P29320 293 ) and non-complimenting cells ( A549 ) , with the viral expression in P29320 293 cells being 2.1 times greater than for A549 cells . 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 . 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 . Characterization of a thyroiditis-inducing thyroglobulin-specific T-cell clone restricted by the H-2 molecule of a low responder mouse strain . We established a thyroglobulin ( Tg ) -specific , thyroiditis-inducing T-cell clone , B12G , from B6C3F1 mice by the immunization of mouse Tg with lipopolysaccharide ( LPS ) from Klebsiella strain LEN ( O3: P04264 ) . B12G was Thy-1.2+ , CD3+ , P01730 + , P05107 + , and CD8- , and could transfer thyroiditis to recipient mice after in vitro stimulation with mouse or bovine Tg . Histological examination showed severe thyroiditis with predominant infiltrations of polymorphonuclear cells ; few mononuclear cells were observed . B12G proliferated in response to bovine , mouse , porcine , and rat Tg in the presence of irradiated spleen cells , but did not respond to chicken or human Tg . H-2b , a low-responder haplotype of experimental autoimmune thyroiditis , governed the response of the clone to Tg . B12G produced interleukin-4 ( P05112 ) and P05231 , but not P60568 or interferon-gamma ( P01579 ) , on stimulation with mouse Tg . These findings were different from characteristics of previously reported Tg-specific T-cell clones from high-responder mice in terms of epitope specificity and cytokine production pattern , raising the possibility that the specificities and functions of T cells involved in the development of autoimmune thyroiditis in low-responder mice differ from those in high responders . 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 . Targeting of plasmid DNA-lipoplexes to cells with molecules anchored via a metal chelator lipid . BACKGROUND : The ability to deliver plasmid DNA ( pDNA ) to specific cells in vivo is crucial for achieving efficient targeted transfection with nonviral vectors . We previously used stealth liposomes containing the chelator lipid 3 ( nitrilotriacetic acid ) -ditetradecylamine ( NTA(3)-DTDA ) to target delivery of antigen and cytokines to immune cells in vivo . In the present study , we utilized liposomes containing NTA(3)-DTDA and the ionizable aminolipid 1,2-dioleoyl-3-dimethyl-ammonium-propane ( DODAP ) to incorporate pDNA into complexes for targeting to cells . METHODS : Liposomes containing DODAP , NTA(3)-DTDA and helper lipids were acidified ( pH 5.5 ) and mixed with pDNA to form complexes . These lipoplexes were neutralized and engrafted with DB00117 -tagged molecules for targeting to extracellular receptors . Targeted transfection efficiency was assessed using the enhanced green fluorescent protein reporter gene . RESULTS : Initial transfections of P29320 -293 cells using a DB00117 -tagged peptide ( P24752 ) related to the DB00125 -rich motif of HIV-1 TAT protein resulted in a low transfection efficiency ( < 2.5 % ) . Optimization of the lipid formulation and use of an endosome-destabilizing peptide and inhibitor of DNase II , increased transfection approximately 20-fold . These lipoplexes are approximately 250 nm in diameter , and transfection efficiencies were : approximately 50 % for P29320 -293 cells targeted with lipoplexes containing pEGFP-N1 and engrafted with P24752 , and 30-40 % for HepG2 cells targeted with lipoplexes engrafted with a peptide specific for the P15692 receptor Flt-1 . CONCLUSIONS : The results show that DODAP-containing lipoplexes incorporating NTA(3)-DTDA enable the engraftment of targeting molecules and the effective targeting of pDNA to cells in serum-containing media , resulting in efficient transgene expression . The strategy may provide a convenient approach for targeting pDNA to cells in vivo in therapeutic applications . Absence of clinically important Q12809 channel blockade by three compounds that inhibit phosphodiesterase 5 -- sildenafil , tadalafil , and vardenafil . Compounds that inhibit phosphodiesterase 5 ( O76074 ) have been developed for the treatment of erectile dysfunction . Because men with erectile dysfunction frequently have comorbid cardiovascular disease , they may have limited cardiac repolarization reserve and be at risk of arrhythmia if treated with medications that prolong ventricular repolarization . The human ether-a-go-go related gene ( Q12809 ) channel is important for repolarization in human myocardium and is a common target for drugs that prolong the QT interval . We studied the ability of three compounds that inhibit O76074 -- sildenafil , tadalafil , and vardenafil -- to block the Q12809 channel . Using a whole cell variant of the patch-clamp method , the Q12809 current was measured in a stably transfected human embryonic kidney cell line expressing the Q12809 channel . The compounds produced dose-dependent reductions in Q12809 current amplitude over a concentration range of 0.1 to 100 microM . The IC50 values were 12.8 microM for vardenafil and 33.3 microM for sildenafil . Because the maximum soluble concentration of tadalafil ( 100 microM ) produced only a 50.9 % inhibition of the Q12809 current amplitude , the IC50 value for tadalafil could not be determined with the Hill equation . DB00820 had the weakest capacity to block the Q12809 channel , producing a 50.9 % blockade at the maximum soluble concentration ( 100 microM ) , compared with 86.2 % for vardenafil ( 100 microM ) and 75.2 % for sildenafil ( 100 microM ) . In conclusion , the concentrations of the O76074 inhibitors required to evoke a 50 % inhibition of the Q12809 current were well above reported therapeutic plasma concentrations of free and total compound . None of the three compounds was a potent blocker of the Q12809 channel . 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 . [ Effect of the genetic markers Kell( P04264 ) , P1 , Km(1) , Tf ( C < B < D ) , AK and 6- P52209 on the outcome of paternity cases ] . This statistical analysis of the results of 288 paternity cases is a contribution to the discussion of those blood group systems to be selected for the basis of paternity expertise in the Federal Republic of Germany . When typing 22 blood-group systems in 288 one-man cases , we found exclusions in 101 ( 35.07 % ) of them . In only 83 ( 44.39 % ) of the 187 cases with nonexclusions did the resulting EM value correspond to the verbal predicate : " paternity practically proved. " The results of the systems of factors Kell( P04264 ) , Tf(C,B,D) , AK and 6- P52209 had the smallest rate of exclusion constellations and only inferior influence on the resulting EM values . Replacing them by isoelectric focusing of the systems P36871 , Tf , Gc , Pi and P00747 ( plasminogen ) seems to be reasonable . The factors P1 and Km(1) proved more favorable for the results of paternity cases . [ 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 . The protease-activated receptor-3 ( PAR-3 ) can signal autonomously to induce interleukin-8 release . Protease-activated receptors ( PARs ) play a clear role in the burst of inflammatory reactions and immune responses . However , for PAR-3 , the most elusive member of the PAR family , the functional role is still largely unclear . It has been claimed that PAR-3 does not signal autonomously , although the wide expression of human PAR-3 indicates its important physiological roles . We demonstrate that in P29320 -293 cells , stably transfected with human PAR-3 , thrombin induced calcium signaling , P10145 gene expression and P10145 release . We confirmed this finding using human lung epithelial and human astrocytoma cells that express endogenous PAR-3 . Moreover , thrombin exposure of P29320 -293 cells resulted in P27361 /2 activation coinciding with P10145 release . The effects of thrombin were not dependent on P25116 activation , as confirmed by P25116 gene silencing . Thus , we propose that PAR-3 is able to signal autonomously to induce P10145 release mediated by P27361 /2 phosphorylation , which contributes actively to inflammatory responses . Disulfide-dependent protein folding is linked to operation of the vitamin K cycle in the endoplasmic reticulum . A protein disulfide isomerase- Q9BQB6 redox enzyme complex appears to be responsible for vitamin P04264 2,3-epoxide reduction . Gamma-carboxylation of vitamin K-dependent proteins is dependent on formation of reduced vitamin P04264 ( Vit.K1H2 ) in the endoplasmic reticulum ( ER ) , where it works as an essential cofactor for gamma-carboxylase in post-translational gamma-carboxylation of vitamin K-dependent proteins . Vit.K1H2 is produced by the warfarin-sensitive enzyme vitamin K 2,3-epoxide reductase ( Q9BQB6 ) of the vitamin K cycle that has been shown to harbor a thioredoxin-like CXXC center involved in reduction of vitamin P04264 2,3-epoxide ( Vit.K > O ) . However , the cellular system providing electrons to the center is unknown . Here data are presented that demonstrate that reduction is linked to dithiol-dependent oxidative folding of proteins in the ER by protein disulfide isomerase ( P07237 ) . Oxidative folding of reduced RNase is shown to trigger reduction of Vit.K > O and gamma-carboxylation of the synthetic gamma-carboxylase peptide substrate FLEEL . In liver microsomes , reduced RNase-triggered gamma-carboxylation is inhibited by the P07237 inhibitor bacitracin and also by small interfering RNA silencing of P07237 in P29320 293 cells . Immunoprecipitation and two-dimensional SDS-PAGE of microsomal membrane proteins demonstrate the existence of a Q9BQB6 enzyme complex where P07237 and Q9BQB6 appear to be tightly associated subunits . We propose that the P07237 subunit of the complex provides electrons for reduction of the thioredoxin-like CXXC center in Q9BQB6 . We can conclude that the energy required for gamma-carboxylation of proteins is provided by dithiol-dependent oxidative protein folding in the ER and thus is linked to de novo protein synthesis . Suppression of synaptic plasticity by cerebrospinal fluid from anti- DB01221 receptor encephalitis patients . The functional effects of cerebrospinal fluid ( P04141 ) from patients with anti- DB01221 receptor ( NMDAR ) encephalitis on the NMDAR-mediated synaptic plasticity were evaluated by using mouse hippocampus slices . Anti-NMDAR antibody detection system was established by immunostaining recombinant NMDAR heteromers expressed in P29320 cell culture as well as native NMDARs in cultured hippocampal neurons . Under a complete blind manner for the clinical information , P04141 and sera collected from 36 pre-diagnosed patients were tested for anti-NMDAR antibodies . With this test , thirteen patients were diagnosed as anti-NMDAR encephalitis . P04141 positive for anti-NMDAR antibodies suppressed induction of long-term potentiation ( LTP ) at Schaffer collateral- P00915 synapses in mouse hippocampal slices . LTP induction was not suppressed by P04141 collected from herpes simplex virus ( HSV ) encephalitis or non-encephalitis control patients . Antibody absorption with NMDAR-expressing P29320 cell culture reversed the suppression of LTP by anti-NMDAR encephalitis patients ' P04141 , confirming that anti-NMDAR antibodies suppressed LTP . The present experiments firmly support the proposal that the anti-NMDAR encephalitis autoantibody is responsible for cognitive disorders like amnesia accompanying this disease . Identification of a potent inverse agonist at a constitutively active mutant of human Q9H244 receptor . Human platelets express two P2Y receptors : G(q)-coupled P2Y(1) , and G(i)-coupled P2Y(12) . Both P2Y(1) and P2Y(12) are ADP receptors on human platelets and are essential for ADP-induced platelet aggregation that plays pivotal roles in thrombosis and hemostasis . Numerous constitutively active G protein-coupled receptors have been described in natural or recombinant systems , but in the P2Y receptors , to date , no constitutive activity has been reported . In our effort to identify G protein coupling domains of the human platelet ADP receptor , we constructed a chimeric hemagglutinin-tagged human P2Y(12) receptor with its C terminus replaced by the corresponding part of human P2Y(1) receptor and stably expressed it in Chinese hamster ovary- P04264 cells . It is interesting that the chimeric P2Y(12) mutant exhibited a high level of constitutive activity , as evidenced by decreased DB02527 levels in the absence of agonists . The constitutive activation of the chimeric P2Y(12) mutant was dramatically inhibited by pertussis toxin , a G(i) inhibitor . The constitutively active P2Y(12) mutant retained normal responses to 2-methylthio-ADP , with an EC(50) of 0.15 +/- 0.04 nM . The constitutively active P2Y(12) mutant caused Akt phosphorylation that was abolished by the addition of pertussis toxin . Pharmacological evaluation of several P2Y(12) antagonists revealed ( E ) -N-[1-[7-(hexylamino)-5-(propylthio)-3H-1,2,3-triazolo-[4,5-d]-pyrimidin-3-yl]-1,5,6-trideoxy-beta-d-ribo-hept-5-enofuranuronoyl]-l-aspartic acid ( AR-C78511 ) as a potent P2Y(12) inverse agonist and 5'-adenylic acid , N-[2-(methylthio)ethyl]-2-[(3,3,3-trifluoropropyl)thio]- , monoanhydride with (dichloromethylene)bis [ phosphonic acid ] ( AR-C69931MX ) as a neutral antagonist . In conclusion , this is the first report of a cell line stably expressing a constitutively active mutant of human platelet P2Y(12) receptor and the identification of potent inverse agonist . 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 . 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 . 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 . 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 . 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 . Suppression of dendritic cell maturation by Trichinella spiralis excretory/secretory products . Evidence from experimental studies indicates that during chronic infections with certain helminth species a regulatory network is induced that can down-modulate not only parasite-induced inflammation but also reduce other immunopathologies such as allergies and autoimmune diseases . The mechanisms however , and the molecules involved in this immunomodulation are unknown . Here , we focus on the effect of Trichinella spiralis excretory/secretory antigens ( TspES ) on the innate immune response by studying the effect of TspES on DC maturation in vitro . Bone marrow-derived DC from BALB/c mice were incubated with TspES either alone or in combination with LPS derived from two different bacteria . As indicators of DC maturation , the cytokine production ( IL-1alpha , P05231 , P22301 , IL-12p70 and P01375 ) and the expression of various surface molecules ( MHC-II , P25942 , P33681 and P42081 ) were measured . Results indicate that while TspES alone did not change the expression of the different surface molecules or the cytokine production , it completely inhibited DC maturation induced by Escherichia coli LPS ( E. coli LPS ) . In contrast , DC maturation induced by LPS from another bacterium , Neisseria meningitidis , was not affected by TspES . These results were confirmed using O00206 / Q9Y6Y9 / P08571 transfected P29320 293 cells . In conclusion , T. spiralis ES antigens lead to suppression of DC maturation but this effect depends on the type of LPS used to activate these cells . Genomic comparison of Escherichia coli P04264 strains isolated from the cerebrospinal fluid of patients with meningitis . Escherichia coli is a major cause of enteric/diarrheal diseases , urinary tract infections , and sepsis . E. coli P04264 is the leading gram-negative organism causing neonatal meningitis , but the microbial basis of E. coli P04264 meningitis is incompletely understood . Here we employed comparative genomic hybridization to investigate 11 strains of E. coli P04264 isolated from the cerebrospinal fluid ( P04141 ) of patients with meningitis . These 11 strains cover the majority of common O serotypes in E. coli P04264 isolates from P04141 . Our data demonstrated that these 11 strains of E. coli P04264 can be categorized into two groups based on their profile for putative virulence factors , lipoproteins , proteases , and outer membrane proteins . Of interest , we showed that some open reading frames ( ORFs ) encoding the type III secretion system apparatus were found in group 2 strains but not in group 1 strains , while ORFs encoding the general secretory pathway are predominant in group 1 strains . These findings suggest that E. coli P04264 strains isolated from P04141 can be divided into two groups and these two groups of E. coli P04264 may utilize different mechanisms to induce meningitis . Leptin modulates the intrinsic excitability of AgRP/ P01303 neurons in the arcuate nucleus of the hypothalamus . The hypothalamic arcuate nucleus ( Q5SW96 ) is a brain region critical for regulation of food intake and a primary area for the action of leptin in the CNS . In lean mice , the adipokine leptin inhibits neuropeptide Y ( P01303 ) and agouti-related peptide ( AgRP ) neuronal activity , resulting in decreased food intake . Here we show that diet-induced obesity in mice is associated with persistent activation of P01303 neurons and a failure of leptin to reduce the firing rate or hyperpolarize the resting membrane potential . However , the molecular mechanism whereby diet uncouples leptin 's effect on neuronal excitability remains to be fully elucidated . In P01303 neurons from lean mice , the Kv channel blocker 4-aminopyridine inhibited leptin-induced changes in input resistance and spike rate . Consistent with this , we found that Q5SW96 P01303 neurons have a large , leptin-sensitive delayed rectifier K(+) current and that leptin sensitivity of this current is blunted in neurons from diet-induced obese mice . This current is primarily carried by Kv2-containing channels , as the Kv2 channel inhibitor stromatoxin-1 significantly increased the spontaneous firing rate in P01303 neurons from lean mice . In P29320 cells , leptin induced a significant hyperpolarizing shift in the voltage dependence of Kv2.1 but had no effect on the function of the closely related channel Kv2.2 when these channels were coexpressed with the long isoform of the leptin receptor LepRb . Our results suggest that dynamic modulation of somatic Kv2.1 channels regulates the intrinsic excitability of P01303 neurons to modulate the spontaneous activity and the integration of synaptic input onto these neurons in the Q5SW96 . Atrial natriuretic peptide binds to P01160 - Q96GN5 receptors in neuroblastoma cells or is degraded extracellularly at the DB00133 - DB00120 bond . P01160 - Q96GN5 receptors for atrial natriuretic peptide ( P01160 ) showed the following rank order of affinity in intact human neuroblastoma cells NB-OK-1 : human P01160 -(99-126) approximately human P01160 -(102-126) approximately rat P01160 -(99-126) ( P04264 17-32 pM ) > human P01160 -(103-126) > porcine brain natriuretic peptide ( DB04899 ) . Analogues truncated at the C-terminal extremity or devoid of a disulphide bridge , such as rat P01160 -(103-123) , rat C- P01160 -(102-121) , rat P01160 -(111-126) , rat P01160 -(99-109) and rat [desCys105,Cys121] P01160 -(104-126) and chicken P23582 , were not recognized . The occupancy of these high affinity P01160 - Q96GN5 receptors led to marked cyclic GMP accumulation in the presence of 3-isobutyl 1-methylxanthine . An ectoenzymic activity , partly shed in the incubation medium , provoked the stepwise release of DB00120 - DB00125 -[125I] DB00135 , DB00125 -[125I] DB00135 and [125I] DB00135 from rat [125I] P01160 -(99-126) , at an optimal pH of 7.0 . Its inhibition by 1,10-phenanthroline , DB00974 and bacitracin but not by thiorphan suggests the contribution of at least one neutral metalloendopeptidase , distinct from EC 3.4.24.11 , for which P01160 showed high affinity . 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 .

MH_train_1129

MH_train_1129

MH_train_1129

interacts_with DB08895?

multiple_choice

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 . Mechanisms of Nattokinase in protection of cerebral ischemia . In vivo , the level of cyclic DB00640 Monophosphate ( DB02527 ) and the pathway of the Janus Kinase1/Signal Transducers and Activators of Transcription1 ( P23458 / P42224 ) were studied . In vitro , the Ca(2+) mobilization in human platelet stimulated by thrombin was observed . In addition , vasomotion of vascular smooth muscle was measured by adding DB00761 or norepinephrine(NE) under the Ca(2+) contained bath solutions . The effect induced by NE in the presence of N-nitro-L-arginine methyl ester ( L-NAME ) or indometacin ( Indo ) was also detected . At last , the levels of tissue plasminogen activator ( t-PA ) and P00747 activator inhibitor-1 ( P05121 ) in cultured supernatans in Human umbilical vein endothelial cells ( Huvecs ) were measured by means of ELISA kit . Results showed that Nattokinase ( NK ) significantly increased the DB02527 level , activated the signal passage of P23458 / P42224 in injured part and inhibited remarkably the rise of platelet intracellular Ca(2+) ( [Ca(2+)]i ) in human platelet . Furthermore , NK relaxed rat thoracic aortic artery in the dose-dependent manner and in the endothelium dependent manner and its effect could be attenuated by L-NAME . Also , the secretion of t-PA and P05121 were reduced stimulated by Adr on Huvecs . These data indicated that the neuroprotective effect of NK was associated with its antiplatelet activity by elevating DB02527 level and attenuating the calcium release from calcium stores ; with its anti-apoptotic effect through the activation of P23458 / P42224 pathway ; with its relaxing vascular smooth muscle by promoting synthesis and release of NO , reducing ROC calcium ion influx and with its protection on endothelial cells through increasing fibrinolytic activity and facilitating spontaneous thrombolysis . 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 . 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 . 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 . 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 . The O60674 / P40763 and mitochondrial pathways are essential for quercetin nanoliposome-induced P13671 glioma cell death . The formulation of quercetin nanoliposomes ( QUE-NLs ) has been shown to enhance QUE antitumor activity in P13671 glioma cells . At high concentrations , QUE-NLs induce necrotic cell death . In this study , we probed the molecular mechanisms of QUE-NL-induced P13671 glioma cell death and examined whether QUE-NL-induced programmed cell death involved Bcl-2 family and mitochondrial pathway through P40763 signal transduction pathway . Downregulation of Bcl-2 and the overexpression of Bax by QUE-NL supported the involvement of Bcl-2 family proteins upstream of P13671 glioma cell death . In addition , the activation of O60674 and P40763 were altered following exposure to QUE-NLs in P13671 glioma cells , suggesting that QUE-NLs downregulated Bcl-2 mRNAs expression and enhanced the expression of mitochondrial mRNAs through P40763 -mediated signaling pathways either via direct or indirect mechanisms . There are several components such as ROS , mitochondrial , and Bcl-2 family shared by the necrotic and apoptotic pathways . Our studies indicate that the signaling cross point of the mitochondrial pathway and the O60674 / P40763 signaling pathway in P13671 glioma cell death is modulated by QUE-NLs . In conclusion , regulation of O60674 / P40763 and ROS-mediated mitochondrial pathway agonists alone or in combination with treatment by QUE-NLs could be a more effective method of treating chemical-resistant glioma . P52333 activity is necessary for phosphorylation of cytosolic phospholipase A2 and prostaglandin E2 synthesis by macrophages infected with Francisella tularensis live vaccine strain . Francisella tularensis , the causative agent of tularemia , modulates the host immune response to gain a survival advantage within the host . One mechanism of immune evasion is the ability of F. tularensis to induce the synthesis of the small lipid mediator prostaglandin E2 ( DB00917 ) , which alters the host T cell response making the host more susceptible to Francisella growth . DB00917 is synthesized by a tightly regulated biosynthetic pathway following stimulation . The synthesis of DB00917 begins with the liberation of arachidonic acid ( AA ) from membrane phospholipids by cytosolic phospholipase A2 ( P47712 ) . AA is subsequently converted to the unstable intermediate PGH2 by cyclooxygenase-2 ( P35354 ) , and PGH2 undergoes an isomerization reaction to generate DB00917 . Our objective was to identify F. tularensis-activated host signaling pathways that regulate the activity of the enzymes in the DB00917 -biosynthetic pathway . In this study , we show that P47712 , p38 mitogen-activated protein kinase ( MAPK ) , and P52333 ( P52333 ) signaling are necessary for F. tularensis-induced DB00917 production . Inhibition of P52333 activity reduced the phosphorylation of P47712 and P35354 protein levels . In addition , P52333 regulates P47712 phosphorylation independent of transcription . Moreover , p38 MAPK activity is required for F. tularensis-induced P35354 protein synthesis , but not for the phosphorylation of P47712 . This research highlights a unique signaling axis in which P52333 and p38 MAPK regulate the activity of multiple enzymes of the DB00917 -biosynthetic pathway in macrophages infected with F. tularensis . 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 . 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 . 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 . 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 . Modulation of leukocyte infiltration and phenotype in microporous tissue engineering scaffolds via vector induced P22301 expression . Biomaterial scaffolds are central to many tissue engineering strategies as they create a space for tissue growth and provide a support for cell adhesion and migration . However , biomaterial implantation results in unavoidable injury resulting in an inflammatory response , which can impair integration with the host and tissue regeneration . Toward the goal of reducing inflammation , we investigated the hypothesis that a lentiviral gene therapy-based approach to localized and sustained P22301 expression at a scaffold could modulate the number , relative proportions , and cytokine production of infiltrating leukocyte populations . Flow cytometry was used to quantify infiltration of six leukocyte populations for 21 days following implantation of P00747 scaffolds into intraperitoneal fat . Leukocytes with innate immune functions ( i.e. , macrophages , dendritic cells , neutrophils ) were most prevalent at early time points , while T lymphocytes became prevalent by day 14 . Reporter gene delivery indicated that transgene expression persisted at the scaffold for up to 28 days and macrophages were the most common leukocyte transduced , while transduced dendritic cells expressed the greatest levels of transgene . P22301 delivery decreased leukocyte infiltration by 50 % relative to controls , increased macrophage P22301 expression , and decreased macrophage , dendritic cell , and P01730 T cell IFN-γ expression . Thus , P22301 gene delivery significantly decreased inflammation following scaffold implant into the intraperitoneal fat , in part by modulating cytokine expression of infiltrating leukocytes . 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 . 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 . 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 . 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 . Aberrant collagenase expression in chronic idiopathic myelofibrosis is related to the stage of disease but not to the O60674 mutation status . Bone marrow fibrosis in chronic idiopathic myelofibrosis ( cIMF ) most likely represents an imbalance between synthesis and turnover of collagen fibers . Because the JAK- P35610 signaling pathway is involved in the regulation of genes encoding matrix metalloproteinases ( MMPs ) , we examined the expression of MMPs , their tissue inhibitors ( TIMPs ) , and collagen types in relation to the O60674 status ( V617F mutation versus wild-type ) in cIMF ( n = 64 ) . Whereas no correlation was found between the O60674 status and MMP gene products , there was an evident association with the stage of disease . Membrane type 1-MMP ( P50281 ) was overexpressed by up to 80-fold in advanced stages that progressed to fibrosis ( P < 0.001 ) , and megakaryocytes and endothelial cells were unmasked as the major cellular source . By contrast , a significantly higher expression of neutrophil collagenase ( P22894 ) was encountered in the prefibrotic stages of cIMF ( P < 0.001 ) . Altogether , the stepwise progress of myelofibrosis in cIMF was associated with expression of a defined subset of target genes as shown in sequential trephine biopsies of cIMF patients . We conclude that the expression of matrix-modeling genes in cIMF is not influenced by the O60674 mutation status but is predominantly related to the stage of disease . 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 . 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 . [ 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 . 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 . P60568 regulates expression of C- O75444 in human P01730 T cells . Blockade of IL-2R with humanized anti-CD25 Abs , such as daclizumab , inhibits Th2 responses in human T cells . Recent murine studies have shown that P60568 also plays a significant role in regulating Th2 cell differentiation by activated P42229 . To explore the role of activated P42229 in the Th2 differentiation of primary human T cells , we studied the mechanisms underlying P60568 regulation of C- O75444 expression . Chromatin immunoprecipitation studies revealed that P60568 induced P42229 binding to specific sites in the C- O75444 promoter . These sites corresponded to regions enriched for markers of chromatin architectural features in both resting P01730 and differentiated Th2 cells . Unlike P05231 , P60568 induced C- O75444 expression in P01730 T cells with or without prior TCR stimulation . TCR-induced C- O75444 expression was significantly inhibited by treatment with daclizumab or a P52333 inhibitor , R333 . Furthermore , P60568 and P05231 synergistically induced C- O75444 expression in TCR-activated T cells , suggesting functional cooperation between these cytokines . Finally , both TCR-induced early P05112 mRNA expression and P05112 cytokine expression in differentiated Th2 cells were significantly inhibited by IL-2R blockade . Thus , our findings demonstrate the importance of P60568 in Th2 differentiation in human T cells and support the notion that IL-2R-directed therapies may have utility in the treatment of allergic disorders . 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 . Characterization of the insulin-signaling pathway in lacrimal and salivary glands of rats . PURPOSE : P01308 has been acknowledged as a mediator of several physiological events in lacrimal and salivary glands . We investigated the presence of insulin receptors and of insulin-induced autophosphorylation of the insulin receptor and activation of elements involved in the early steps of insulin signaling in lacrimal and salivary glands of rats . METHODS : Lacrimal and salivary glands of Wistar rats were removed and processed for immunohistochemistry using anti-insulin receptor and anti- DB01277 receptor antibodies . The activation of insulin receptors following insulin treatment , and the involvement of insulin receptor substrates-1 and -2 , Shc , O60674 and P35610 -1 , were analyzed by immunoprecipitation , followed by SDS-PAGE and immunoblotting of rat lacrimal and salivary glands after exposure to insulin . RESULTS : P01308 and DB01277 receptors were present in rat lacrimal and salivary glands and were located predominantly in the cytoplasm and plasma membrane . Functional studies demonstrated that insulin induced a dose-dependent phosphorylation of the insulin receptor , IGF-1R , insulin receptor substrates-1 and -2 , Shc , and P35610 -1 . In rats with streptozotocin-induced diabetes mellitus there was a significant reduction in insulin-induced insulin receptor and P35610 -1 phosphorylation in the lacrimal gland but not in the salivary gland ; there was no influence on Shc phosphorylation in either tissue . CONCLUSIONS : The present results indicate that insulin and DB01277 receptors are expressed in lacrimal and salivary glands , and that insulin can induce the phosphorylation of its receptor and activate elements involved in the early steps of insulin signaling in both tissues . 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 . 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 . 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 . 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 . A cytosolic protein-tyrosine phosphatase P18031 specifically dephosphorylates and deactivates prolactin-activated STAT5a and STAT5b . P01236 ( PRL ) plays a central and crucial role in the regulation of milk protein gene expression in mammary epithelial cells . PRL binding to its cognate receptor leads to receptor dimerization and activation of the tyrosine kinase O60674 ( O60674 ) , associated with the membrane-proximal , intracellular domain of the receptor . In turn , O60674 phosphorylates and activates P42229 , a member of the signal transducers and activators of transcription ( P35610 ) family . We have recently reported that 16 different protein-tyrosine phosphatases ( PTP ) were expressed in lactating mouse mammary gland and mammary epithelial cells ( Aoki , N. , Kawamura , M. , Yamaguchi-Aoki , Y. , Ohira , S. , and Matsuda , T. ( 1999 ) J. Biochem. ( Tokyo ) 125 , 669-675 ) . We investigated the involvement of each PTP in PRL signaling . Among the 12 phosphatases including Q06124 examined , a cytosolic phosphatase P18031 was found to specifically dephosphorylate STAT5a and STAT5b in transfected COS7 and in vitro . Nuclear translocation of STAT5a and STAT5b was largely inhibited upon overexpression of P18031 . The PRL-dependent transcriptional activation of the beta-casein gene promoter was also inhibited by P18031 . Furthermore , retrovirus-mediated overexpression of P18031 resulted in dephosphorylation of endogenous P42229 and down-regulation of beta-casein gene expression in mammary epithelial COMMA-1D cells when the cells were treated with lactogenic hormones . Endogenous tyrosine-phosphorylated P42229 proteins in mammary epithelial COMMA-1D cells as well as tyrosine-phosphorylated STAT5a and STAT5b expressed in COS7 cells were co-precipitated by substrate-trapping mutants of recombinant P18031 . These results strongly suggest that P18031 dephosphorylates PRL-activated STAT5a and STAT5b , thereby negatively regulating PRL-mediated signaling pathway . The role of P01375 in insulin resistance . P01308 resistance is an important component of the metabolic syndrome associated with obesity . Early-stage insulin-resistance and related mild glucose intolerance may be compensated by increased insulin secretion . When combined with impaired insulin secretion , insulin resistance plays an important role in type 2 diabetes ( 1 ) . P01308 -resistance is also associated with a variety of pathological conditions , including trauma , infection , and cancer . Obesity and type 2 diabetes are the most common metabolic diseases in Western societies , together affecting as much as half of the adult population ( 2 ) . The prevalence of these conditions is not only high , but continues to increase . We have only recently come to appreciate the role of fat , especially visceral fat , as an endocrine organ . Visceral fat is the source of a number of substances which might play a role in the development of insulin resistance . Among the latter are tumor necrosis factor-alpha ( P01375 ) , adiponectin , P05231 , resistin and free fatty acids . This review will discuss the regulation of insulin responses by P01375 and evidence supporting the hypothesis that over expression of P01375 plays a role in the pathophysiology of insulin resistance . Human adipose tissue macrophages are enhanced but changed to an anti-inflammatory profile in obesity . OBJECTIVE : Adipose tissue ( AT ) macrophages are increased in obesity and associated with low grade inflammation . We aimed to characterize the phenotype of AT macrophages in humans in relation to obesity and insulin resistance . DESIGN : Gene-expression levels of general macrophage markers ( P34810 and P08571 ) , proinflammatory markers/M1 ( P01375 -α , P13500 , and P05231 ) , and anti-inflammatory markers/M2 ( Q86VB7 , CD206 , and P22301 ) were determined by RT-PCR in subcutaneous AT samples from lean and obese subjects . P01308 resistance was determined by HOMA-IR . RESULTS : All the macrophage markers were elevated in the AT from obese compared to lean subjects ( P < 0.001 ) . To determine the phenotype of the macrophages the level of P08571 was used to adjust the total number of macrophages . The relative expression of Q86VB7 and P22301 was elevated , and P01375 -α and P05231 were reduced in AT from obese subjects ( all P < 0.05 ) . In a multivariate regression analysis Q86VB7 was the only macrophage marker significantly associated with HOMA-IR ( β : 0.57 ; P < 0.05 ) . CONCLUSION : Obesity is associated with elevated numbers of macrophages in the AT . Unexpectedly , the macrophages change phenotype by obesity , with a preponderance of M2 and a decrement of M1 markers in AT from obese subjects . Moreover , Q86VB7 was the only macrophage marker associated with HOMA-IR after multiple adjustments . Pharmacogenetics of antipsychotic-induced weight gain : review and clinical implications . Second-generation antipsychotics ( SGAs ) , such as risperidone , clozapine and olanzapine , are the most common drug treatments for schizophrenia . SGAs presented an advantage over first-generation antipsychotics ( FGAs ) , particularly regarding avoidance of extrapyramidal symptoms . However , most SGAs , and to a lesser degree FGAs , are linked to substantial weight gain . This substantial weight gain is a leading factor in patient non-compliance and poses significant risk of diabetes , lipid abnormalities ( that is , metabolic syndrome ) and cardiovascular events including sudden death . The purpose of this article is to review the advances made in the field of pharmacogenetics of antipsychotic-induced weight gain ( AIWG ) . We included all published association studies in AIWG from December 2006 to date using the Medline and ISI web of knowledge databases . There has been considerable progress reaffirming previous findings and discovery of novel genetic factors . The P28335 and leptin genes are among the most promising , and new evidence suggests that the P14416 , P01375 , P60880 and P32245 genes are also prominent risk factors . Further promising findings have been reported in novel susceptibility genes , such as P21554 , P08183 , ADRA1A and Q9Y5U4 . More research is required before genetically informed , personalized medicine can be applied to antipsychotic treatment ; nevertheless , inroads have been made towards assessing genetic liability and plausible clinical application . Tanshinone IIA inhibits constitutive P40763 activation , suppresses proliferation , and induces apoptosis in rat P13671 glioma cells . P40763 ( P40763 ) is usually constitutively activated in a variety of malignancies . Thus , P40763 may be a promising target for treatment of tumor cells . Recently , Tanshinone IIA ( Tan IIA ) , a major active constituent from the root of Salvia miltiorrhiza Bunge , was reported to have apoptosis inducing effects on a large variety of cancer cells . In this study , we evaluate the anti-proliferation and apoptosis inducing effects of Tan IIA on P13671 glioma cells . Cell growth and proliferation were measured by MTT assay , cell apoptosis was observed by flow cytometry and DNA-fragmentation analysis . Further more , we investigated inhibitory effects of Tan IIA on P40763 activity and its downstream targets : Bcl-XL , cyclin D1 . Alteration of P40763 activity was examined by measuring their DNA binding activity and tyrosine phosphorylation . Changes in the expression levels of Bcl-XL and cyclin D1 were examined by Western blot analysis . We found that the cellular growth were inhibited and cell apoptosis were observed after the treatment with Tan IIA . The P40763 activity was significantly reduced by Tan IIA parallel with a significant attenuation of expression of Bcl-XL and cyclin D1 . These results suggest that Tan IIA may serve as an effective adjunctive reagent in the treatment of glioma for its targeting of constitutive P40763 signaling . [ 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 . 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 . Genetic association between P43250 /β-arrestin 2 and dopamine supersensitivity psychosis in schizophrenia . BACKGROUND/AIM : Dopamine supersensitivity psychosis ( P15924 ) , clinically characterized by unstable and severe psychosis or tardive dyskinesia and often categorized as treatment-resistant schizophrenia , is promoted by long-term antipsychotic treatment . An upregulation of the dopamine D2 receptor caused by antipsychotic(s) is involved in the development of P15924 . The present study explored the potential roles of P43250 ( P43250 ) and β-arrestin 2 ( P32121 ) that are involved in the trafficking of P14416 in patients with P15924 . METHODS : We conducted a genetic association study of P43250 / P32121 between the patients with P15924 episodes [ P15924 (+) group : N=108 ] and the patients without P15924 (-) episodes [ P15924 (-) group : N=169 ] from the total group of patients ( N=333 ) . Based on the patients ' treatment history , a P15924 episode was defined as withdrawal psychosis , developed tolerance to antipsychotic effect , and tardive dyskinesia ( the remaining 56 patients were excluded due to insufficient information ) . RESULTS : The results revealed that none of the allelic or genotyping distributions of five single nucleotide polymorphisms ( SNPs ) of P43250 and three SNPs of P32121 showed any significant difference between the P15924 (+) and P15924 (-) groups . CONCLUSION : The results suggest that the SNP analyses of these two molecules fail to classify patients into the potential clinical subtype of P15924 (+) or P15924 (-) group . However , since P43250 and P32121 are surely involved in dopamine D2 receptor metabolism , further studies based on prospective observations of the onset of P15924 under specific antipsychotic treatments are needed . Drosophila Answers to Q13148 Proteinopathies . Initially implicated in the pathogenesis of P13569 and HIV-1 transcription , nuclear factor Q13148 was subsequently found to be involved in the origin and development of several neurodegenerative diseases . In 2006 , in fact , it was reported for the first time the cytoplasmic accumulation of Q13148 in ubiquitin-positive inclusions of P35858 and FTLD patients , suggesting the presence of a shared underlying mechanism for these diseases . Today , different animal models of Q13148 proteinopathies are available in rodents , nematodes , fishes , and flies . Although these models recapitulate several of the pathological features found in patients , the mechanisms underpinning the progressive neuronal loss observed in Q13148 proteinopathies remain to be characterized . Compared to other models , Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies . At present , the development of Q13148 -related Drosophila models has further strengthened the hypothesis that both Q13148 " loss-of-function " and " gain-of-function " mechanisms can contribute to disease . The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies , along with the information they have generated , towards a better understanding of the mechanisms underlying Q13148 proteinopathies . 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 . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . 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 . Human and rodent pancreatic beta-cells express P05112 receptors and P05112 protects against beta-cell apoptosis by activation of the PI3K and JAK/ P35610 pathways . Secretion of pro-inflammatory cytokines is associated with loss of pancreatic beta-cell viability and cell death . P05112 ( interleukin-4 ) has been reported to mediate a protective effect against the loss of pancreatic beta-cells , and P05112 receptors have been found in rat pancreatic beta-cells at both the RNA and the protein level . The aim of the present study was to investigate P05112 receptor expression in human islet cells and to examine the signalling pathways by which P05112 exerts its effects using the rat beta-cell lines , BRIN-BD11 and P01308 -1E . By means of immunohistochemistry , it was demonstrated that P05112 receptors are present on human islet cells . Using a flow cytometric method for evaluating cell death , it was confirmed that incubating beta-cells with P05112 attenuated cell death induced by IL-1beta and interferon-gamma by approx . 65 % . This effect was abrogated by the presence of the PI3K ( phosphoinositide 3-kinase ) inhibitor , wortmannin , suggesting that activation of the PI3K pathway is involved . In support of this , Western blotting revealed that incubation of cells with P05112 resulted in increased phosphorylation of Akt ( also called protein kinase B ) , a downstream target of PI3K . Increased tyrosine phosphorylation of P42226 ( signal transducer and activator of transcription 6 ) also occurred in response to P05112 and a selective P52333 ( P52333 ) inhibitor reduced the cytoprotective response . Both effects were prevented by overexpression of the tyrosine phosphatase , PTP-BL ( protein tyrosine phosphatase-BL ) . We conclude that P05112 receptors are functionally competent in pancreatic beta-cells and that they signal via PI3K and JAK/ P35610 pathways . These findings may have implications for future therapeutic strategies for the management of diabetes . [ 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 . Gene network analysis leads to functional validation of pathways linked to cancer cell growth and survival . Hepatocellular carcinoma ( HCC ) represents one of the most frequently diagnosed human cancers ; however , there are currently few treatment alternatives to surgical resection . In this study we performed bioinformatic analysis of previously published transcriptomic data in order to characterize liver specific networks , including biological functions , signaling pathways and transcription factors , potentially dysregulated in HCC . By incorporating specific signaling inhibitors into real-time proliferation assays using HepG2 cells , we then validated these in silico results . We found that G protein subunits Gi/G0 , protein kinase C , Mek1/2 , and Erk1/2 ( P42/44 ) , P23458 , Q07869 and NFκB p65 subunit were the major signaling molecules required for survival and proliferation of human HCC cell lines . We also found that these pathways regulate the expression of key hepatic transcription factors involved in cell differentiation , such as P49715 , P18146 , Q08050 and PPARs . By combining bioinformatic and functional analyses , major signaling pathways related to tumorigenicity in HCC are revealed , thereby elucidating potential targets for drug therapies . Human gray matter : feasibility of single-slab 3D double inversion-recovery high-spatial-resolution MR imaging . The purpose of this study was to develop and prospectively evaluate the feasibility of a single-slab three-dimensional ( 3D ) double inversion-recovery , or P30518 , sequence for magnetic resonance imaging at 1.5 T . The study was approved by the local ethics committee , and informed consent was obtained from six healthy control subjects ( one woman , five men ; age range , 26-47 years ) and two patients with multiple sclerosis ( one woman , aged 39 ; one man , aged 56 ) . Gray matter ( GM ) -only images were obtained by selectively suppressing cerebrospinal fluid ( P04141 ) and white matter ( WM ) signals . Whole-brain high-spatial-resolution 3D images ( 1.2 x 1.2 x 1.3 mm ) were acquired within 10 minutes . Cortical and deep GM structures were clearly delineated from WM and P04141 , and there were regional differences in GM signal intensity . No flow artifacts from blood or P04141 were observed . These GM images with high spatial resolution are suitable to identify cortical pathologic conditions and can potentially be used for segmentation purposes to determine cortical thickness or volume . 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 . Dopamine receptors and psychiatric drug treatment . The established antipsychotic drugs act mainly by antagonizing dopamine mediated synaptic transmission in the brain . Increase in the rate of production of dopamine metabolites as well as the firing rate of dopamine-containing neurons can be interpreted as compensatory responses to an interruption of synaptic transmission at dopamine nerve terminals . The demonstration of involvement of limbic and cortical mechanisms in the antipsychotic activity of neuroleptic drugs is far more difficult than the involvement of nigro-striatal and tubero-infundibular mechanisms in the neurological and neuroendocrine effects of these drugs . Application of radioreceptor techniques to dopamine research has supported the findings obtained by other neuropsychopharmacological research techniques , providing more direct evidence of dopamine receptor blockade by neuroleptic drugs . Further research is needed especially in studying the nature of the time-dependent adaptive changes at the receptor sites as well as the differences between the different dopamine projections and neural systems in the brain . The different subtypes of dopamine receptors in the brain , currently called D1 and D2 dopamine receptors , seem to be parallel , although in many respects independently-acting regulatory systems . P14416 -selective antagonists such as sulpiride seem to cause selective D2 receptor up-regulation . P01236 secretion seems to be regulated by D2 dopamine receptors . The exact physiological role of D1 dopamine receptors as well as the clinical consequences of selective D1 antagonism is not known . DB00391 and clozapine are examples of atypical neuroleptic compounds that have quite different profile of action , the former having strong and selective antidopaminergic action , the latter combining a number of non- dopaminergic mechanisms with rather slight effects on dopamine receptors. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . Molecular dissection of human oncostatin M-mediated signal transductions through site-directed mutagenesis . The binding of oncostatin M ( OM ) to type I and type II receptor complexes elicits various biological responses by activating MEK/ P29323 and JAK/ P35610 signaling pathways . Some OM effects are clinically desirable such as reducing hyperlipidemia through the activation of hepatic P01130 transcription , a downstream event of P29323 activation . The OM pro-inflammatory responses via induction of acute phase protein gene expression have been associated with P35610 activation . In this study , by conducting site-directed mutagenesis , bioassays and molecular modeling we have defined 4 OM residues that are differently involved in the activation of P29323 or P35610 signaling pathway in HepG2 cells . We show that mutation of Lys163 to alanine totally abolished OM-mediated signaling , possibly because such mutation causes the disruption of a stabilizing H-bond pattern at the OM interface with receptors . G120A mutation equally impaired activations of P29323 and P35610 signaling pathways also by impairing the OM/cognate protein interactions . Interestingly , mutations of Gln20 and Asn123 differentially affected OM signaling through the two pathways . Q20A and N123A retained strong activity in inducing P29323 phosphorylation but they showed diminished ability in activating P42224 and P40763 . We further showed that mutations at Gln20 and Asn123 reduced OM induction of inflammatory gene fibrinogen-beta to a greater extent than that of P01130 gene . The mutation of Asn123 is directly related to local structural modification at site 3 of OM . Collectively these results provide a structural basis of OM-mediated signaling and suggest a potential to improve OM therapeutic properties via structural modification . The cleaved cytoplasmic tail of polycystin-1 regulates Src-dependent P40763 activation . P98161 ( PC1 ) mutations result in proliferative renal cyst growth and progression to renal failure in autosomal dominant polycystic kidney disease ( ADPKD ) . The transcription factor P40763 ( signal transducer and activator of transcription 3 ) was shown to be activated in cyst-lining cells in ADPKD and Q15139 mouse models and may drive renal cyst growth , but the mechanisms leading to persistent P40763 activation are unknown . A proteolytic fragment of PC1 corresponding to the cytoplasmic tail , PC1-p30 , is overexpressed in ADPKD . Here , we show that PC1-p30 interacts with the nonreceptor tyrosine kinase Src , resulting in Src-dependent activation of P40763 by tyrosine phosphorylation . The PC1-p30-mediated activation of Src/ P40763 was independent of JAK family kinases and insensitive to the P40763 inhibitor suppressor of cytokine signaling 3 . Signaling by the P01133 receptor ( P00533 ) or DB02527 amplified the activation of Src/ P40763 by PC1-p30 . Expression of PC1-p30 changed the cellular response to DB02527 signaling . In the absence of PC1-p30 , DB02527 dampened P00533 - or P05231 -dependent activation of P40763 ; in the presence of PC1-p30 , DB02527 amplified Src-dependent activation of P40763 . In the polycystic kidney ( PCK ) rat model , activation of P40763 in renal cystic cells depended on vasopressin receptor 2 ( P30518 ) signaling , which increased DB02527 levels . Genetic inhibition of vasopressin expression or treatment with a pharmacologic P30518 inhibitor strongly suppressed P40763 activation and reduced renal cyst growth . These results suggest that PC1 , via its cleaved cytoplasmic tail , integrates signaling inputs from P00533 and DB02527 , resulting in Src-dependent activation of P40763 and a proliferative response . 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 . 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 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 . 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 . [ 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 . Intracerebroventricular administration of insulin and glucose inhibits the anorectic action of leptin in rats . Obese individuals with glucose intolerance present with high serum levels of glucose , insulin , and leptin . These substances are potent inhibitors of feeding in the brain . Obese subjects still present with over-feeding despite elevation of the above factors . To elucidate the mechanism of this paradox , the effects of insulin and glucose on the anorectic action of leptin in the hypothalamus were examined . Adult male Sprague-Dawley rats ( weighing 285-320 g ) were pretreated with intracerebroventricular injection of insulin , glucose , or saline , followed by leptin ( 7.5 microg ) or phosphate-buffered saline ( PBS ) injection into the third cerebral ventricle ( icv ) . The cumulative food intakes were measured 24 hr after leptin icv . The tyrosine phosphorylation of signal transducer and activator transcription factor 3 ( P40763 ) in the hypothalamus was determined by Western blotting . In rats pretreated with saline and stimulated with leptin ( saline/LEPTIN group ) , food intake diminished to about 50 % of that of the saline/PBS group ( P < 0.005 ) . Food intake in the insulin/LEPTIN group was significantly higher compared with the saline/LEPTIN group ( P < 0.005 ) and reached the level seen in the saline/PBS group . Similar data were obtained in glucose pretreatment experiments . P01308 and glucose icv resulted in reduction of leptin-induced P40763 tyrosine phosphorylation compared with saline . Infusion of insulin and glucose icv did not alter peripheral blood glucose levels in all groups . High insulin or glucose levels in the brain could result in leptin resistance as manifested by food intake , which is probably due to the attenuation of P40763 phosphorylation downstream the leptin receptor . The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . Differential regulation of cystic fibrosis transmembrane conductance regulator by interferon gamma in mast cells and epithelial cells . P13569 ( P13569 ) is a P13569 in epithelial cells ; recently , we identified it in mast cells . Previous work that we confirmed showed that interferon gamma ( IFNgamma ) down-regulated P13569 expression in epithelial cells ( T84 ) , but by contrast , we found that IFNgamma up-regulated P13569 mRNA and protein expression in rat and human mast cells . IFNgamma up-regulation of P13569 in mast cells was inhibited by p38 and extracellular signal-regulated kinase ( P29323 ) kinase inhibitors but not a Janus tyrosine kinase (JAK)2 inhibitor , whereas in T84 cells IFNgamma-mediated down-regulation of P13569 was O60674 -dependent and P29323 - and p38-independent . Furthermore , IFNgamma down-regulation of P13569 in T84 epithelial cells was P42224 -dependent , but up-regulation of P13569 in mast cells was P42224 -independent . Thus , differential regulatory pathways of P13569 expression in mast cells and epithelial cells exist that depend upon either p38/ P29323 or JAK/ P35610 pathways , respectively . Surprisingly , IFNgamma treatment of mast cells inhibited Cl(-) efflux , in contrast to up-regulation of P13569 /mRNA and protein expression . However , down-regulation of Cl(-) flux correlated with IFNgamma-mediated inhibition of mediator secretion . This and other work suggests that the effect of IFNgamma on P13569 expression in mast cells is important for their function . 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 . 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 . Induction of autophagy biomarker Q12983 requires a O60674 / P40763 and P50281 signaling interplay in Concanavalin-A-activated U87 glioblastoma cells . Plant lectins have been considered as possible anti-tumor drugs because of their property to induce autophagic cell death . Given that expression of membrane type-1 matrix metalloproteinase ( P50281 ) has been found to regulate expression of the autophagy biomarker Bcl-2/adenovirus E1B 19kDa interacting protein 3 ( Q12983 ) , we sought to investigate possible signaling interplay mechanisms between P50281 and Q12983 in Concanavalin-A ( ConA ) lectin-activated U87 glioblastoma cells . ConA induced acidic vacuole organelle formation as well as Q12983 and P50281 gene and protein expressions , whereas only Q12983 expression was dose-dependently inhibited by the O60674 tyrosine kinase inhibitor AG490 suggesting a requirement for some P35610 -mediated signaling . Gene silencing of P50281 and of P40763 abrogated ConA-induced P40763 phosphorylation and Q12983 expression . Correlative analysis shows that P40763 signaling events occur downstream from P50281 induction . Overexpression of a full length P50281 recombinant protein led to increased Q12983 gene and protein expressions . The cytoplasmic domain of P50281 was also found necessary for transducing P40763 phosphorylation . Among P23458 , O60674 , P52333 , and P29597 , only O60674 gene silencing abrogated ConA 's effects on P50281 and Q12983 gene and protein expressions . Our study elucidates how P50281 signals autophagy , a process which could contribute to the chemoresistance phenotype in brain cancer cells . 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 .

MH_train_1130

MH_train_1130

MH_train_1130

interacts_with DB00030?

multiple_choice

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 . 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 . 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 . G protein-coupled receptor kinase 2 mediates endothelin-1-induced insulin resistance via the inhibition of both Galphaq/11 and insulin receptor substrate-1 pathways in 3T3- Q9NUQ9 adipocytes . G protein-coupled receptor kinases ( GRKs ) regulate seven-transmembrane receptors ( 7TMRs ) by phosphorylating agonist-activated 7TMRs . Recently , we have reported that P25098 can function as a negative regulator of insulin action by interfering with G protein-q/11 alpha-subunit ( Galphaq/11 ) signaling , causing decreased glucose transporter 4 ( P14672 ) translocation . We have also reported that chronic endothelin-1 ( ET-1 ) treatment leads to heterologous desensitization of insulin signaling with decreased tyrosine phosphorylation of insulin receptor substrate ( P41252 ) -1 and Galphaq/11 , and decreased insulin-stimulated glucose transport in 3T3- Q9NUQ9 adipocytes . In the current study , we have investigated the role of P25098 in chronic ET-1-induced insulin resistance . P01308 -induced P14672 translocation was inhibited by pretreatment with ET-1 for 24 h , and we found that this inhibitory effect was rescued by microinjection of anti- P25098 antibody or P25098 short interfering RNA . We further found that P25098 mediates the inhibitory effects of ET-1 by two distinct mechanisms . Firstly , adenovirus-mediated overexpression of either wild-type ( WT ) - or kinase-deficient ( KD ) - P25098 inhibited Galphaq/11 signaling , including tyrosine phosphorylation of Galphaq/11 and cdc42-associated phosphatidylinositol 3-kinase activity . Secondly , ET-1 treatment caused DB00133 / DB00156 phosphorylation of P35568 and P35568 protein degradation . Overexpression of KD- P25098 , but not WT- P25098 , inhibited ET-1-induced serine 612 phosphorylation of P35568 and restored activation of this pathway . Taken together , these results suggest that P25098 mediates ET-1-induced insulin resistance by 1 ) inhibition of Galphaq/11 activation , and this effect is independent of P25098 kinase activity , and 2 ) P25098 kinase activity-mediated P35568 serine phosphorylation and degradation . Feedback upregulation of P21860 ( ErbB3 ) expression and activity attenuates antitumor effect of PI3K inhibitors . We examined the effects of an inhibitor of PI3K , DB05240 , against human breast cancer cell lines with constitutive PI3K activation . Treatment with DB05240 resulted in dose-dependent inhibition of cell growth and levels of pAKT and pS6 , signal transducers in the PI3K/AKT/TOR pathway . In P04626 -overexpressing cells , inhibition of PI3K was followed by up-regulation of expression and phosphorylation of multiple receptor tyrosine kinases , including P21860 . Knockdown of FoxO1 and FoxO3a transcription factors suppressed the induction of P21860 , InsR , P08069 , and P21802 mRNAs upon inhibition of PI3K . In P04626 (+) cells , knockdown of P21860 with siRNA or cotreatment with the P04626 inhibitors trastuzumab or lapatinib enhanced DB05240 -induced cell death and inhibition of pAKT and pS6 . DB00072 and lapatinib each synergized with DB05240 for inhibition of pAKT and growth of established BT474 xenografts . These data suggest that PI3K antagonists will inhibit AKT and relieve suppression of receptor tyrosine kinase expression and their activity . Relief of this feedback limits the sustained inhibition of the PI3K/AKT pathway and attenuates the response to these agents . As a result , PI3K pathway inhibitors may have limited clinical activity overall if used as single agents . In patients with P04626 -overexpressing breast cancer , PI3K inhibitors should be used in combination with P04626 / P21860 antagonists . 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 . 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 ) Purification of rat P13671 glioma plasma membranes by affinity partitioning . We have studied the feasibility of purifying rat P13671 glioma plasma membranes by a phase partitioning approach . The purification procedure involves cell homogenization and fractionation with an aqueous two-phase polymer system followed by selective affinity purification of plasma membranes by a wheat germ agglutinin-coupled polymer system . We demonstrate that the two-phase affinity partitioning technique is a simple and efficient method of isolating cell plasma membranes with high purity and yield . Furthermore , the isolated plasma membranes retain their functional integrity , as shown by the high-affinity insulin-like growth factor-I ( P05019 ) binding capacity of P05019 receptors . 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 . 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 . 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 . 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 . 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 .

MH_train_1131

MH_train_1131

MH_train_1131

interacts_with DB00091?

multiple_choice

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 . 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 . 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 . 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 . 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 . 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 . 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 . DB11320 reduces susceptibility to natural killer cells via down-regulation of P26718 ligands on human monocytic leukaemia THP-1 cells . Natural killer ( NK ) group 2D ( P26718 ) is a key activating receptor expressed on NK cells , whose interaction with ligands on target cells plays an important role in tumorigenesis . However , the effect of histamine on P26718 ligands on tumour cells is unclear . Here we showed that human monocytic leukaemia THP-1 cells constitutively express MHC class I-related chain A ( Q29983 ) and Q9BZM6 on their surface , and incubation with histamine reduced the expression in a dose-dependent and time-dependent manner as assessed by flow cytometry . Interferon-γ augmented the surface expression of the P26718 ligands , and this augmentation was significantly attenuated by histamine . The histamine H1 receptor ( P35367 ) agonist 2-pyridylethylamine and P25021 agonist dimaprit down-regulated the expression of P26718 ligands , and activation of P35367 and P25021 signalling by A23187 and forskolin , respectively , had the same effect , indicating that the histamine-induced down-regulation of P26718 ligands is mediated by P35367 and P25021 . Quantitative reverse transcription-PCR showed that mRNA levels of the P26718 ligands and relevant microRNAs were not significantly changed by histamine . DB11320 down-regulated the surface expression of endoplasmic reticulum protein 5 , and inhibition of matrix metalloproteinases did not impair this down-regulation , indicating that proteolytic shedding was not involved . Instead , pharmacological inhibition of protein transport and proteasome abrogated it , and histamine enhanced ubiquitination of Q29983 . Furthermore , histamine treatment significantly reduced susceptibility to NK cell-mediated cytotoxicity . These results suggest that histamine down-regulates P26718 ligands through the activation of an P35367 - and P25021 -mediated ubiquitin-proteasome pathway and consequently reduces susceptibility to NK cells . 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 . 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 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 . 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 . 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 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . P01375 -alpha-induced cyclooxygenase-2 expression via sequential activation of ceramide-dependent mitogen-activated protein kinases , and O15111 1/2 in human alveolar epithelial cells . The role of Q8TCB0 /42 mitogen-activated protein kinase ( MAPK ) , p38 , and c-Jun NH(2)-terminal kinase ( JNK ) in tumor necrosis factor ( P01375 ) -alpha-induced cyclooxygenase ( P36551 ) -2 expression was studied in NCI-H292 epithelial cells . P01375 -mediated P35354 expression and P35354 promoter activity were inhibited by the MAPK kinase inhibitor PD98059 or the p38 inhibitor SB203580 . Treatment of cells for 10 min with P01375 resulted in activation of Q8TCB0 /42 MAPK , p38 , and JNK . P06681 -ceramide ( a cell-permeable ceramide analog ) , bacterial neutral sphingomyelinase ( Smase ; an enzyme that degrades sphingomyelin to ceramide ) , and N-oleoylethanolamine ( a ceramidase inhibitor ) all induced activation of MAPKs , P35354 expression , nuclear factor ( NF ) -kappaB DNA-protein binding , and P35354 promoter activity . The inactive analog , dihydro- P06681 -ceramide , had no effect . SMase- or P06681 -ceramide-induced P35354 expression and P35354 promoter activity were also inhibited by PD98059 or SB203580 . Glutathione , a neutral SMase inhibitor , attenuated P01375 - or SMase-induced activation of MAPKs , P35354 expression , and P35354 promoter activity . P01375 - or P06681 -ceramide-induced P35354 promoter activity was inhibited by the dominant negative mutant of extracellular signal-regulated kinase 2 , p38 , JNK , O15111 (IKK)1 , or O14920 . IKK activity was stimulated by either P01375 or P06681 -ceramide , and these effects were inhibited by PD98059 or SB203580 . All these results suggest that , in NCI-H292 epithelial cells , activation of MAPKs by ceramide contributes to the P01375 signaling that occurs downstream of neutral SMase activation and results in the stimulation of O15111 /2 , and NF-kappaB in the P35354 promoter , followed by initiation of P35354 expression . 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 . DB06287 : a safety and efficacy review . INTRODUCTION : The vascular endothelial growth factor ( P15692 ) pathway and the mammalian Target of DB00877 ( P42345 ) represent the most frequently exploited targets in renal cell carcinoma ( RCC ) . DB06287 is an inhibitor of P42345 , and is a unique ester derivative of sirolimus , a macrocyclic lactone , with improved pharmaceutical properties , including stability and solubility . DB06287 binds to the cytoplasmic protein P62942 , and the complex binds and inhibits P42345 . AREAS COVERED : This review summarizes the clinical findings and safety of temsirolimus in RCC patients . EXPERT OPINION : A Phase III clinical trial has demonstrated that temsirolimus has statistically significant advantages over treatment with IFN-α in RCC patients with poor prognosis , in terms of OS ( overall survival ) , PFS ( progression-free survival ) , and tumor response . Median OS was improved 49 % compared to IFN-α , and median PFS was approximately doubled . It is now considered the standard for RCC patients with poor prognostic features . The possibility that this agent is useful in metastatic non-clear cell carcinoma patients has also been suggested by a subset analysis of the pivotal Phase III trial . Studies in untreated favorable and intermediate risk clear cell and refractory mRCC patients are required . 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 . 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 . Cord blood stem-cell-derived dendritic cells generate potent antigen-specific immune responses and anti-tumour effects . The aim of the present study was to investigate whether CBSCs [ ( umbilical ) cord blood stem cells ] can be a new source of DCs ( dendritic cells ) , which can generate more potent antigen-specific immune responses and anti-tumour effects . CBSCs and PBMCs ( peripheral blood mononuclear cells ) were collected , cultured and differentiated into DCs . Surface markers , secreting cytokines , antigen-presentation activity , antigen-specific cell-mediated immunity and cytotoxic killing effects induced by these two DC origins were evaluated and compared . CBSCs were expanded ~17-fold by ex vivo culture . The expression of surface markers in CBSC-derived DCs were higher than those in PBMC-derived DCs treated with LPS ( lipopolysaccharide ) . The CBSC-derived DCs mainly secreted IL (interleukin)-6 , P22301 and P01375 ( tumour necrosis factor ) -α , whereas PBMC-derived DCs mainly secreted P05113 and IFN (interferon)-γ . The CBSC-derived DCs had better antigen-presentation abilities when stimulated with LPS or P01375 -α , induced higher numbers of IFN-γ-secreting antigen-specific CD8+ T-cells , as assessed using an ELISpot ( enzyme-linked immunosorbent spot ) assay , and stimulated more potent antigen-specific CTL ( cytotoxic T-cell ) activities ( P < 0.01 , one-way Q9UNW9 ) . CBSC-derived DCs had quicker and greater P29323 ( extracellular-signal-regulated kinase ) and Akt phosphorylation , and weaker p38 phosphorylation , than PBMC-derived DCs when stimulated with LPS . In conclusion , CBSC-derived DCs have the ability to induce stronger antigen-specific immunity and more potent anti-tumour effects and therefore could be a good source of DCs for use in DC-based cancer vaccines and immunotherapy . P25021 activation exacerbates myocardial ischemia/reperfusion injury by disturbing mitochondrial and endothelial function . There is evidence that P25021 blockade improves ischemia/reperfusion ( I/R ) injury , but the underlying cellular mechanisms remain unclear . DB11320 is known to increase vascular permeability and induce apoptosis , and these effects are closely associated with endothelial and mitochondrial dysfunction , respectively . Here , we investigated whether activation of the histamine H2 receptor ( P25021 ) exacerbates myocardial I/R injury by increasing mitochondrial and endothelial permeability . Serum histamine levels were measured in patients with coronary heart disease , while the influence of P25021 activation was assessed on mitochondrial and endothelial function in cultured cardiomyocytes or vascular endothelial cells , and myocardial I/R injury in mice . The serum histamine level was more than twofold higher in patients with acute myocardial infarction than in patients with angina or healthy controls . In neonatal rat cardiomyocytes , histamine dose-dependently reduced viability and induced apoptosis . Mitochondrial permeability and the levels of p- P27361 /2 , Bax , p- Q9UIK4 , and caspase 3 were increased by P25021 agonists . In cultured human umbilical vein endothelial cells ( HUVECs ) , P25021 activation increased p- P27361 /2 and p-moesin levels and also enhanced permeability of HUVEC monolayer . All of these effects were abolished by the P25021 blocker famotidine or the P29323 inhibitor U0126 . After I/R injury or permanent ischemia , the infarct size was reduced by famotidine and increased by an P25021 agonist in wild-type mice . In P25021 KO mice , the infarct size was smaller ; myocardial p- P27361 /2 , p- Q9UIK4 , and mitochondrial Bax were downregulated . These findings indicate that P25021 activation exaggerates myocardial I/R injury by promoting myocardial mitochondrial dysfunction and by increasing cardiac vascular endothelial permeability . A novel mechanism of autophagic cell death in dystrophic muscle regulated by Q99572 receptor large-pore formation and HSP90 . Q99572 is an DB00171 -gated ion channel , which can also exhibit an open state with a considerably wider permeation . However , the functional significance of the movement of molecules through the large pore ( LP ) and the intracellular signaling events involved are not known . Here , analyzing the consequences of Q99572 activation in primary myoblasts and myotubes from the Dmd(mdx) mouse model of Duchenne muscular dystrophy , we found DB00171 -induced Q99572 -dependent autophagic flux , leading to P42574 - P55210 -independent cell death . Q99572 -evoked autophagy was triggered by LP formation but not Ca(2+) influx or P28482 - P27361 phosphorylation , 2 canonical Q99572 -evoked signals . Phosphoproteomics , protein expression inference and signaling pathway prediction analysis of Q99572 signaling mediators pointed to P54652 and HSP90 proteins . Indeed , specific HSP90 inhibitors prevented LP formation , LC3-II accumulation , and cell death in myoblasts and myotubes but not in macrophages . Pharmacological blockade or genetic ablation of p2rx7 also proved protective against DB00171 -induced death of muscle cells , as did inhibition of autophagy with 3-MA . The functional significance of the Q99572 LP is one of the great unknowns of purinergic signaling . Our data demonstrate a novel outcome -- autophagy -- and show that molecules entering through the LP can be targeted to phagophores . Moreover , we show that in muscles but not in macrophages , autophagy is needed for the formation of this LP . Given that Q99572 -dependent LP and HSP90 are critically interacting in the DB00171 -evoked autophagic death of dystrophic muscles , treatments targeting this axis could be of therapeutic benefit in this debilitating and incurable form of muscular dystrophy . The functional proteomics analysis of P15692 -treated human epithelial ovarian cancer cells . Vascular endothelial growth factor ( P15692 ) , one of the most important angiogenic factor , can impact the tumor cell proliferation and invasion , but the mechanism remains unclear . This study is to investigate the key proteins which may play an important role in the P15692 -induced progress of ovarian cancer cells . The total protein from HO-8910 cells was separated by two-dimensional electrophoresis ( 2-DE ) , and differentially expressed proteins were identified by matrix-assisted laser desorption and ionization time-of-flight tandem mass spectrometry ( MALDI-TOF MS ) and PDQuest image analysis software . Furthermore , real-time PCR , Western blot , and immunocytochemistry were also used to confirm different expression levels of differential proteins . Morphological changes and invasion capability were evaluated by electron microscope and Matrigel invasion assay , respectively . The highly reproducible and well-resolved 2-DE patterns of both HO-8910/ P15692 and HO-8910 cells were acquired . A total of 17 expressed differential proteins were identified , 8 proteins were upregulated ( P60709 , TIM , P30101 , P07237 , Q13561 , KIC17 , SIAS , and KIC10 ) and 9 downregulated ( KIC18 , P11021 , CAPG , P62937 , ROA2 , P02545 , EZRI , Q16186 , and ENOA ) . Ultrastructure of P15692 -treated group showed more malignant characteristic compared with control group , an obvious increase in the number of cells penetrating the Matrigel membrane in P15692 -treated group ( P < 0.05 ) . These results suggested that P15692 could impact ovarian cancer 's malignant progression by regulating expression of associated proteins . 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 . 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 . 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 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . P62937 and P62942 interact with P25490 and alter its transcriptional activity . P25490 is a zinc finger transcription factor with unusual structural and functional features . In a yeast two-hybrid screen , two cellular proteins , cyclophilin A ( CyPA ) and FK506-binding protein 12 ( P62942 ) , interacted with P25490 . These interactions are specific and also occur in mammalian cells . DB00091 and FK506 efficiently disrupt the P25490 -CyPA and P25490 - P62942 interactions . Overexpression of human CyPA and P62942 have different effects on P25490 -regulated transcription with these effects being promoter-dependent . These results suggest that immunophilins may be mediators in the functional role of P25490 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Optimal reference genes for qPCR in resting and activated human NK cells -- Flow cytometric data correspond to qPCR gene expression analysis . Natural killer cells ( NK cells ) are cytotoxic lymphocytes critical to the innate immune system engaged in rapid response against tumor or virus infected cells . After activation NK cells acquire enhanced cytotoxicity and are capable of producing cytokines to stimulate other immune cells . Quantitative PCR ( qPCR ) is a method of choice for gene expression analysis but the usage of reliable reference genes for the normalization process is critical . Commonly used reference genes may vary in expression level between different experimental conditions providing wrong quantitative results of the studied genes ' expression levels . Fourteen potential endogenous control genes were analyzed by qPCR method in NK-92 cell line that shows characteristics of human natural killer cells and is often used in studies on biology of NK lymphocytes . NK-92 cells were stimulated with P60568 or P01375 for 2 , 24 or 72 h . Results were analyzed with RefFinder , a program which enables evaluation and screening of reference genes and integrates the currently available major computational programs ( Genorm , Normfinder , BestKeeper and Delta Ct ) . The most stable gene in activated and non-activated NK cells was P61769 , followed by IPO-8 and P04406 and the least stable were P00492 , P62937 and P62910 . The normalization process was performed on P04179 gene and the results of qPCR experiments were confirmed by flow cytometry . The flow cytometric data corresponded to the results of qPCR gene expression analysis performed for the reference genes qualified by RefFinder as the most stable . 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 . Endogenous expression of histamine H1 receptors functionally coupled to phosphoinositide hydrolysis in P13671 glioma cells : regulation by cyclic AMP . 1. The effects of histamine receptor agonists and antagonists on phospholipid hydrolysis in rat-derived P13671 glioma cells have been investigated . 2 . P35367 -stimulation caused a concentration-dependent increase in the accumulation of total [ 3H ] -inositol phosphates in cells prelabelled with [ 3H ] -myo-inositol . The rank order of agonist potencies was histamine ( EC50 = 24 microM ) > N alpha-methylhistamine ( EC50 = 31 microM ) > 2-thiazolylethylamine ( EC50 = 91 microM ) . 3 . The response to 0.1 mM histamine was antagonized in a concentration-dependent manner by the H1-antagonists , mepyramine ( apparent Kd = 1 nM ) and (+)-chlorpheniramine ( apparent Kd = 4 nM ) . In addition , (-)-chlorpheniramine was more than two orders of magnitude less potent than its (+)-stereoisomer . 4 . Elevation of intracellular cyclic AMP accumulation with forskolin ( 10 microM , EC50 = 0.3 microM ) , isoprenaline ( 1 microM , EC50 = 4 nM ) or rolipram ( 0.5 mM ) , significantly reduced the histamine-mediated ( 0.1 mM ) inositol phosphate response by 37 % , 43 % and 26 % respectively . In contrast , 1,9-dideoxyforskolin did not increase cyclic AMP accumulation and had no effect on the phosphoinositide response to histamine . 5 . These data indicate the presence of functionally coupled , endogenous histamine H1 receptors in P13671 glioma cells . Furthermore , the results also indicate that H1 receptor-mediated phospholipid hydrolysis is inhibited by the elevation of cyclic AMP levels in these cells . Postprandial cell defense system responses to meal formulations : stratification through gene expression profiling . SCOPE : Cell defenses regulating homeostatic control of postprandial stress are influenced by interindividual variation , food composition and health status . This study investigates effects of food composition on individual postprandial responses and associations with health . METHODS AND RESULTS : Volunteers ( n = 16 ) consumed four food formulations ( 50 % unsaturated/saturated fat , with/without beetroot extract 10 g/100 g ) on separate occasions . GeXP assay measured whole blood postprandial gene expression profiles of 28 cell defense markers at baseline and postprandial time points 1 , 2 , 4 , 6 , 24 h . Plasma markers of metabolic lipids , hormones , inflammatory cytokines , oxidative stress , and DNA damage/repair were also assessed . SIRT 1 , P55851 , P09601 , P48637 , P35354 , P04637 , CDKN2A , P62937 , O14543 , and P27695 expression profiles revealed distinct stratified subgroups associated with plasma HDLs , P01375 -α and postprandial responses of O14543 , and P62937 . Leptin , P05231 , and DNA strand breaks revealed differing responses to fat type consumed . CONCLUSION : This study demonstrates postprandial immune , inflammatory , redox , metabolic , and DNA repair responses that are largely independent of fat type consumed ( unsaturated/saturated ) or addition of beetroot extract , in apparently healthy individuals . However , postprandial responses can be characterized by regulation of gene expression associated with markers linked to health status and are subject to interindividual variation that can influence postprandial responses . Gene expression results in lipopolysaccharide-stimulated monocytes depend significantly on the choice of reference genes . BACKGROUND : Gene expression in lipopolysaccharide ( LPS ) -stimulated monocytes is mainly studied by quantitative real-time reverse transcription PCR ( RT-qPCR ) using P04406 ( glyceraldehyde 3-phosphate dehydrogenase ) or P60709 ( beta-actin ) as reference gene for normalization . Expression of traditional reference genes has been shown to vary substantially under certain conditions leading to invalid results . To investigate whether traditional reference genes are stably expressed in LPS-stimulated monocytes or if RT-qPCR results are dependent on the choice of reference genes , we have assessed and evaluated gene expression stability of twelve candidate reference genes in this model system . RESULTS : Twelve candidate reference genes were quantified by RT-qPCR in LPS-stimulated , human monocytes and evaluated using the programs geNorm , Normfinder and BestKeeper. geNorm ranked P23284 ( cyclophilin B ) , P61769 ( beta-2-microglobulin ) and P62937 ( cyclophilin A ) as the best combination for gene expression normalization in LPS-stimulated monocytes . Normfinder suggested P20226 ( TATA-box binding protein ) and P61769 as the best combination . Compared to these combinations , normalization using P04406 alone resulted in significantly higher changes of P01375 ( tumor necrosis factor-alpha ) and P22301 ( interleukin 10 ) expression . Moreover , a significant difference in P01375 expression between monocytes stimulated with equimolar concentrations of LPS from N. meningitides and E. coli , respectively , was identified when using the suggested combinations of reference genes for normalization , but stayed unrecognized when employing a single reference gene , P60709 or P04406 . CONCLUSIONS : Gene expression levels in LPS-stimulated monocytes based on RT-qPCR results differ significantly when normalized to a single gene or a combination of stably expressed reference genes . Proper evaluation of reference gene stabiliy is therefore mandatory before reporting RT-qPCR results in LPS-stimulated monocytes . 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 . Autoreactive T cell hybridoma-derived B cell stimulatory factor(s) governing IgA isotype immunoglobulin production by murine Peyer 's patch B cells . In the present study we investigated whether autoreactive T cells derived from murine Peyer 's patches ( PP ) have the capacity to regulate mucosal B cell differentiation to IgA-producing plasma cells in vitro . We also examined whether B cell development is mediated by lymphokines from immunoregulatory T cells - that is , B cell stimulatory factors ( BSF ) and cofactors ( coBSF ) - which include B cell growth factor ( BCGF ) , putative alpha B cell immunoglobulin ( Ig ) heavy chain switch factor ( BSWF alpha ) , and B cell differentiation factor ( BCDF ) , as well as interleukin-2 ( P60568 ) . To this purpose we developed in vitro a variety of BSF ( especially putative BSWF alpha ) -producing autoreactive ( self-class II molecules responsive ) T cell hybridoma cell clones from murine PP , and studied the functional activity of the BSF , especially a putative alpha Ig heavy chain switch ( mu ---- alpha ) factors(s) . These T hybridome cell lines possessed the surface phenotypes of Thy 1.2 , P01730 + , P06127 + and CD8- and produced a variety of BSF , including two kinds of BCGF ( P05113 , and a BCGF that did not require additional costimulators to induce proliferation of preactivated B cells ) , putative BSWF alpha/gamma , BCDF , and/or P60568 . The results strongly support the view that the autologous T cell plays an important role not only in B cell proliferation and terminal maturation , but also in alpha heavy chain switching in PP . This T-B cell interation is mediated at least in part through BSF lymphokines elaborated by the autoreactive T cell , probably activated in situ in the lymphoid tissue microenvironment . P62937 is overexpressed in human pancreatic cancer cells and stimulates cell proliferation through CD147 . BACKGROUND : Although overexpression of cyclophilin A ( CypA ) is associated with several types of cancer , its role in pancreatic cancer has not been studied . In this study the expression of CypA and its receptor CD147 on pancreatic cancer was determined as well as the effect of exogenous CypA on pancreatic cancer cell proliferation . METHODS : The expression of CypA and CD147 in human pancreatic cancer cell lines and tissues was determined with real-time reverse transcriptase polymerase chain reaction ( RT-PCR ) , Western blot , and immunostaining . Cell proliferation in response to CypA was performed by [3H]thymidine incorporation assay . Phosphorylation of MAPK and cytokine secretion profiles in pancreatic cancer cells were determined by using the Bio-Plex phosphoprotein assay and cytokine assay . RESULTS : Pancreatic cancer cell lines expressed significantly higher levels of CypA and CD147 than normal human pancreatic ductal epithelium ( HPDE ) cells . Expression of CypA and CD147 was also substantially higher in human pancreatic adenocarcinoma tissues than those in normal pancreatic tissues . Addition of exogenous CypA significantly stimulated pancreatic cancer cell proliferation in a dose-dependent manner and this effect was effectively blocked by pretreatment with anti-CD147 antibody . In addition , CypA activated P27361 /2 and p38 MAPK signaling pathways and increased the secretion of 2 key cytokines P05113 and Q16552 in Panc-1 cells . CONCLUSIONS : The expression of CypA and CD147 was significantly increased in both pancreatic cancer cell lines and tissues . Exogenous CypA promotes pancreatic cancer cell growth , which may be mediated through the interaction with CD147 and the activation of P27361 /2 and p38 MAPKs . 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 . 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 . 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 .

MH_train_1132

MH_train_1132

MH_train_1132

interacts_with DB00945?

multiple_choice

P10147 - P51681 axis regulates intratumoral accumulation of leukocytes and fibroblasts and promotes angiogenesis in murine lung metastasis process . Metastasis proceeds through interaction between cancer cells and resident cells such as leukocytes and fibroblasts . An i.v. injection of a mouse renal cell carcinoma , Renca , into wild-type mice resulted in multiple metastasis foci in lungs and was associated with intratumoral accumulation of macrophages , granulocytes , and fibroblasts . A chemokine , P10147 , was detected in infiltrating cells and , to a lesser degree , tumor cells , together with an infiltration of leukocytes expressing P51681 , a specific receptor for P10147 . A deficiency of the P10147 or P51681 gene markedly reduced the number of metastasis foci in the lung , and the analysis using bone marrow chimeric mice revealed that both bone marrow- and non-bone marrow-derived cells contributed to metastasis formation . P10147 - and P51681 -deficient mice exhibited a reduction in intratumoral accumulation of macrophages , granulocytes , and fibroblasts . Moreover , intratumoral neovascularization , an indispensable process for metastasis , was attenuated in these gene-deficient mice . Intrapulmonary expression of matrix metalloproteinase ( MMP ) -9 and hepatocyte growth factor ( P14210 ) was enhanced in wild-type mice , and the increases were markedly diminished in P10147 - and P51681 -deficient mice . Furthermore , P14780 protein was detected in macrophages and granulocytes , the cells that also express P51681 and in vitro stimulation by P10147 -induced macrophages to express P14780 . Intratumoral fibroblasts expressed P51681 and P14210 protein . In vitro P10147 stimulated fibroblasts to express P14210 . Collectively , the P10147 - P51681 axis appears to regulate intratumoral trafficking of leukocytes and fibroblasts , as well as P14780 and P14210 expression , and as a consequence to accelerate neovascularization and subsequent metastasis formation . P15692 , P17948 , P35968 and colorectal cancer : assessment of disease risk , tumor molecular phenotype , and survival . Angiogenesis is essential for tumor progression . Vascular endothelial growth factor ( P15692 ) and its receptors 1 ( P17948 ) and 2 ( P35968 ) , have been identified as major mediators of this process . We hypothesized that genetic variation in P17948 ( 38 SNPs ) , P35968 ( 22 SNPS ) , and P15692 ( 11 SNPs ) would be associated with colon and rectal cancer development and survival . Data from a case-control study of 1555 colon cancer cases and 1956 controls and 754 rectal cancer cases and 959 controls were used . An adaptive rank truncation product ( ARTP ) , based on 10,000 permutations , was used to determine the statistical significance of the candidate genes and angiogenesis pathway . Based on ARTP results , P17948 was significantly associated with risk of colon cancer ( P(ARTP) = 0.045 ) and P15692 was significantly associated with rectal cancer ( P(ARTP) = 0.036 ) . After stratifying by tumor molecular subtype , SNP associations observed for colon cancer were : P15692 rs2010963 with CIMP+ colon tumors ; P17948 rs4771249 and rs7987649 with P04637 ; P17948 rs3751397 , rs7337610 , rs7987649 , and rs9513008 and P35968 rs10020464 , rs11941492 , and rs12498529 with MSI+ and CIMP+/ P01116 -mutated tumors . P17948 rs2296189 and rs600640 were associated with CIMP+ rectal tumors and P17948 rs7983774 was associated with P04637 -mutated rectal tumors . Four SNPs in P17948 were associated with colon cancer survival while three SNPs in P35968 were associated with survival after diagnosis with rectal cancer . DB00945 /NSAID use , smoking cigarettes , and BMI modified the associations . These findings suggest the importance of inflammation and angiogenesis in the etiology of colorectal cancer and that genetic and lifestyle factors may be targets for modulating disease risk . 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 . DB00945 -triggered lipoxin in patients treated with aspirin and selective vs. nonselective P35354 inhibitors . AIMS : Cyclooxygenase ( P36551 ) -2 inhibition has been reported to suppress the biosynthesis of the gastroprotective lipoxygenase metabolite 15(R)-epi-lipoxin A(4) , also termed ' aspirin-triggered lipoxin ' ( ATL ) . We tested the hypothesis that the co-administration of aspirin with either the selective P35354 inhibitor celecoxib or the nonselective P36551 inhibitor ibuprofen reduces ATL biosynthesis . METHODS : We measured the urinary excretion of ATL in 24 patients with both ischaemic heart disease and osteoarthritis , chronically treated with aspirin and co-administered celecoxib 200 mg b.i.d. , ibuprofen 600 mg t.i.d. , or placebo for 7 days . RESULTS : Baseline ATL was comparable in the three groups . On days 1 and 7 , 4 h after co-administration of celecoxib or ibuprofen , ATL levels did not show significant variations ( day 1 : 0.24 +/- 0.33 , 0.26 +/- 0.21 and 0.37 +/- 0.22 ng mg(-1) creatinine , respectively ; day 7 : 0.21 +/- 0.13 , 0.35 +/- 0.15 and 0.23 +/- 0.18 ng mg(-1) creatinine , respectively ) . CONCLUSIONS : Neither selective nor nonselective P35354 inhibition appreciably interferes with ATL biosynthesis , suggesting that this mediator is not involved in exacerbating gastrotoxicity by the association of aspirin with P35354 inhibitors . 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 . Platelet activation in patients with colorectal cancer . DB00945 may reduce the risk of colorectal neoplasia at doses similar to those recommended for the prevention of cardiovascular disease . Thus , we aimed to address whether enhanced platelet activation , as assessed by the measurement of the urinary excretion of 11-dehydro-TXB(2) ( a major enzymatic metabolite of TXB(2) ) , occurs in patients with colorectal cancer . In 10 patients with colorectal cancer , the urinary excretion of 11-dehydro-TXB(2) was significantly higher than in 10 controls , matched for sex , age and cardiovascular risk factors [ 1001(205-5571) versus 409(113-984) pg/mg creatinine , respectively , median ( range ) , P < 0.05 ] . The administration of aspirin 50 mg daily for 5 consecutive days to colorectal cancer patients caused a cumulative inhibition of platelet cyclooxygenase ( P36551 ) -1 activity either ex vivo , as assessed by the measurement of serum TXB(2) levels , or in vivo , as assessed by urinary 11-dehydro-TXB(2) excretion . In conclusion , enhanced platelet activation occurs in colorectal cancer patients . Permanent inactivation of platelet P23219 by low-dose aspirin might restore anti-tumor reactivity . Platelet responsiveness to in vitro aspirin is independent of P23219 and P35354 protein levels and polymorphisms . DB00945 's inhibitory effect on platelet function has been shown to be highly heterogeneous . However , due to the considerable individual variation in pharmacokinetics after aspirin intake , it has been difficult to investigate the mechanism of aspirin resistance empirically . Our objective was to examine whether platelet responsiveness to in vitro aspirin treatment could be affected by cyclooxygenase ( P36551 ) -1/2 protein levels in platelets or single-nucleotide polymorphisms ( SNPs ) , which could possibly change specific activity of enzymes and/or aspirin susceptibility . Collagen/epinephrine closure time ( CEPI-CT ) of PFA-100 in blood from 178 healthy males was assessed with/without aspirin . Platelet P23219 protein levels and the sequences of P23219 gene exons were examined in three groups categorized by CEPI-CT : PR ( Poor responders to aspirin ) , 10 people showing the shortest CEPI-CT under aspirin ; GR-High or GR-Low ( good responders to aspirin with high or low platelet basal reactivity ) , 10 people showing CEPI-CT over 300 s under aspirin and having the shortest or longest basal CEPI-CT , respectively . We analyzed the three groups , representing phenotypic extremes , aiming to increase statistical power to investigate the possible relevance of COXs to platelet response to aspirin . Western blot analysis revealed that P23219 was abundantly expressed in platelets at comparable levels among the three groups , whereas P35354 was undetectable . The frequencies of nonsynonymous P23219 /2 SNPs were unlikely to explain the difference in aspirin responsiveness considering the observed genotype frequencies and wide individual variation in platelet response . These results suggest that heterogeneity in platelet responsiveness to in vitro aspirin is independent of P23219 /2 protein levels and SNPs . 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 . 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 . Potential cardiovascular effects of P35354 selective nonsteroidal antiinflammatory drugs . The newly developed nonsteroidal antiinflammatory drugs ( NSAIDs ) that selectively inhibit cyclooxygenase-2 ( P35354 ) , are effective against pain and inflammation and appear to have less gastrointestinal toxicity than conventional NSAIDs . Their P35354 selectivity , however , has raised concerns regarding their cardiovascular safety , since they do not inhibit P23219 , the isoform of the enzyme that is active in thrombosis and vasoconstriction . At this point there is no conclusive evidence that P35354 inhibitors cause ischemic vascular events , because retrospective post hoc analyses conflict one another , and no specific randomized trials have yet been done . Renal effects , edema and hypertension appear to be similar between conventional NSAIDs and P35354 -selective inhibitors . DB00945 is still required for patients with cardiovascular risk who are prescribed a P35354 -selective inhibitor . 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 . Effect of cyclooxygenase-2 inhibitor ( celecoxib ) on the infarcted heart in situ . Several attempts have been made to replace aspirin with compounds without gastric toxicity ; a cyclooxygenase-2 ( P35354 ) inhibitor , celecoxib , and a nitric oxide-aspirin , O43763 -4016 , have been developed for this purpose . This paper compares effects of celecoxib , O43763 -4016 and aspirin on production of prostacyclin ( DB01240 ) and thromboxane A2 ( TXA2 ) and activation of the inducible form of nitric oxide synthase ( P35228 ) in infarcted heart in situ . DB00945 was most effective in reducing myocardial DB01240 synthesis and formation of TXA2 . Myocardial effects of celecoxib resemble those of O43763 -4016 , although the two compounds have different modes of action . The C50T polymorphism of the cyclooxygenase-1 gene and the risk of thrombotic events during low-dose therapy with acetyl salicylic acid . DB00945 prevents thrombotic events by inhibiting platelet cyclooxygenase-1 ( P23219 ) , thus reducing thromboxane A2 formation and platelet aggregation . The C50T polymorphism of P23219 is associated with an impaired inhibition of both thromboxane production and in-vitro platelet aggregation by aspirin . We studied whether this polymorphism is also associated with the risk of clinical thrombotic events in patients using aspirin . We included 496 patients admitted to our Coronary Care Unit for various indications treated with aspirin 80 mg daily . Genotyping for the C50T polymorphism demonstrated that 86.7 % of the patients had the common genotype , and 13.3 % had the variant ( 12.5 % heterozygous , 0.8 % homozygous ) . Baseline variables were well balanced , except that patients with the common genotype more frequently used aspirin prior to admission compared to those patients with the variant genotype . The composite primary endpoint of myocardial infarction , stroke , and/or cardiovascular death occurred in 98 patients ( 19.8 % ) . Myocardial infarction occurred in 9.6 % of patients , stroke in 1.6 % , and cardiovascular death in 12.1 % . The unadjusted hazard ratio ( 95 % CI ) for the primary endpoint for patients with the variant versus the common genotype was 1.07 ( 0.62-1.85 ) , p = 0.8 . The adjusted hazard ratio was 0.86 ( 0.49-1.50 ) , p = 0.6 . In prior laboratory studies the P23219 C50T polymorphism was associated with an impaired inhibitory effect of aspirin on thromboxane production and platelet function . However , in this cohort of patients using low-dose aspirin for secondary prevention the polymorphism was not associated with a higher risk of atherothrombotic events . 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 . 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 . Clinical pharmacology of cyclooxygenase inhibition and pharmacodynamic interaction with aspirin by floctafenine in Thai healthy subjects . DB08976 , a hydroxyquinoline derivative with analgesic properties , is widely used in Thailand and many other countries . The objectives of this study were to evaluate in Thai healthy volunteers : i ) the inhibition of whole blood cyclooxygenase( P36551 )-2 and P23219 activity by floctafenine and its metabolite floctafenic acid in vitro and ex vivo after dosing with floctafenine ; ii ) the possible interference of floctafenine administration with aspirin antiplatelet effects . We performed an open-label , cross-over , 3-period study , on 11 healthy Thai volunteers , who received consecutively floctafenine(200mg/TID) , low-dose aspirin(81mg/daily) or their combination for 4 days , separated by washout periods . DB08976 and floctafenic acid resulted potent inhibitors of P23219 and P35354 in vitro ( floctafenic acid was more potent than floctafenine ) showing a slight preference for P23219 . After dosing with floctafenine alone , whole blood P23219 and P35354 activities were inhibited ex vivo in a time-dependent fashion which paralleled floctafenic acid plasma concentrations . DB00945 alone inhibited profoundly and persistently platelet P23219 activity and AA-induced platelet aggregation throughout 24-h dosing interval which was affected by the co-administration of floctafenine . At 24 h after dosing with aspirin and floctafenine , the inhibition of platelet thromboxane(TX)B2 generation and aggregation were significantly ( P less than 0.05 ) lower than that caused by aspirin alone . Therapeutic dosing with floctafenine profoundly inhibited prostanoid biosynthesis through the rapid conversion to floctafenic acid . DB08976 interfered with the antiplatelet effect of aspirin . Our results suggest that floctafenine should be avoided in patients with cardiovascular disease under treatment with low-dose aspirin . DB00758 and aspirin in cardiovascular medicine : responders or not -- current best available evidence . Dual antiplatelet therapy represents an important advance for patients with established coronary artery disease . It is an important strategy for patients with acute coronary syndromes and those undergoing percutaneous transcatheter coronary interventions . DB00758 effectively inhibits ADP-induced platelet activation and aggregation by selectively and irreversibly blocking the P2Y(12) receptor on the platelet membrane . DB00945 works by irreversibly acetylating the cyclooxygenase ( P23219 ) enzyme , thus suppressing the production of thromboxane A(2) ( TxA(2) ) and inhibiting platelet activation and aggregation . Variable platelet response and potential resistance to therapy has emerged with aspirin and clopidogrel . The definitions of antiplatelet agents variability in responsiveness and nonresponsiveness are discussed . DB00758 and aspirin responsiveness as they are measured in the laboratory by various techniques ( platelet aggregometry and point-of-care assays such as platelet function analyzer [ PFA-100 ] and rapid platelet function assay [ RPFA ] ) are evaluated . The mechanisms responsible for variations in responsiveness to antiplatelet agents such as clinical , cellular and genetic factors are defined . DB00945 and clopidogrel resistance are emerging clinical entities with potentially severe consequences such as myocardial infarction , stroke or death . The therapeutic interventions to deal with nonresponsiveness are reported , although specific recommendations are not clearly established . In the future , routine measurement of platelet function in patients with cardiovascular disease may become the standard of care . Personalized antithrombotic treatment strategies may be determined by ex-vivo measurements that identify critical pathways influencing thrombotic risk in the individual patient . 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 . DB00945 resistance : clinical significance and genetic polymorphism . OBJECTIVE : To determine the prevalence , clinical implications and underlying mechanism of aspirin resistance in Chinese patients . METHODS : Platelet aggregation was determined by light transmission aggregometry ( P01374 ) using four different inducers . Patients were divided into aspirin-resistant ( AR ) , aspirin semi responder ( ASR ) and aspirin-sensitive ( AS ) groups , according to their P01374 results . DB00945 resistance was assessed by thrombo elastography ( TEG , with arachidonic acid [ AA ] or adenosine diphosphate as inducers ) , serum/urinary 11-dehydrothromboxane B2 ( P28845 -TXB2 ) assay , platelet function analyser-100 assay and P16109 assay . Polymorphisms in the prostaglandin endoperoxide synthase 1 ( P23219 ) gene ( A842G , C50T , C22T , G128A , C644A and C714A ) , the P35354 gene ( G765C ) and the integrin β3 ( P05106 ) gene ( C196T ) were examined . RESULTS : The study included 360 aspirin-treated patients and 314 healthy controls . AS patients had significantly lower levels of P28845 -TXB2 than AR and ASR patients , and significantly lower levels of P16109 than AR patients . TEG-AA was more sensitive , specific and consistent than P16109 in detecting aspirin resistance . The frequency of the P35354 G765C mutation was significantly higher in the AR/ASR groups versus the AS group . CONCLUSIONS : TEG-AA was more sensitive , specific and consistent than the P16109 assay for detecting aspirin resistance , and the P35354 G765C mutation may be related to aspirin resistance . A yeast sensor of ligand binding . We describe a biosensor that reports the binding of small-molecule ligands to proteins as changes in growth of temperature-sensitive yeast . The yeast strains lack dihydrofolate reductase ( P00374 ) and are complemented by mouse P00374 containing a ligand-binding domain inserted in a flexible loop . Yeast strains expressing two ligand-binding domain fusions , P62942 - P00374 and estrogen receptor-alpha ( ERalpha ) - P00374 , show increased growth in the presence of their corresponding ligands . We used this sensor to identify mutations in residues of ERalpha important for ligand binding , as well as mutations generally affecting protein activity or expression . We also tested the sensor against a chemical array to identify ligands that bind to P62942 or ERalpha . The ERalpha sensor was able to discriminate among estrogen analogs , showing different degrees of growth for the analogs that correlated with their relative binding affinities ( RBAs ) . This growth assay provides a simple and inexpensive method to select novel ligands and ligand-binding domains . [ 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 . 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 . Nuclear factor-kappaB enhances ErbB2-induced mammary tumorigenesis and neoangiogenesis in vivo . The ( P04626 /Neu ) ErbB2 oncogene is commonly overexpressed in human breast cancer and is sufficient for mammary tumorigenesis in transgenic mice . Nuclear factor ( NF ) -kappaB activity is increased in both human and murine breast tumors . The immune response to mammary tumorigenesis may regulate tumor progression . The role of endogenous mammary epithelial cell NF-kappaB had not previously been determined in immune-competent animals . Furthermore , the role of the NF-kappaB components , p50 and p65 , in tumor growth was not known . Herein , the expression of a stabilized form of the NF-kappaB-inhibiting P25963 protein ( IkappaBalphaSR ) in breast tumor cell lines that express oncogenic ErbB2 inhibited DNA synthesis and growth in both two- and three-dimensional cultures . Either NF-kappaB inhibition or selective silencing of p50 or p65 led to a loss of contact-independent tumor growth in vitro . IkappaBalphaSR reversed the features of the oncogene-induced phenotype under three-dimensional growth conditions . The NF-kappaB blockade inhibited ErbB2-induced mammary tumor growth in both immune-competent and immune-deficient mice . These findings were associated with both reduced tumor microvascular density and a reduction in the amount of vascular endothelial growth factor . The expression of IkappaBalphaSR in breast cancer tumors inhibited angiogenesis . Thus , mammary epithelial cell NF-kappaB activity enhances ErbB2-mediated mammary tumorigenesis in vivo by promoting both growth and survival signaling via the promotion of tumor vasculogenesis . Nuclear factor kappaB ( NF-kappaB ) pathway as a therapeutic target in rheumatoid arthritis . Rheumatoid arthritis ( RA ) is a chronic inflammatory disease characterized by persistent joint swelling and progressive destruction of cartilage and bone . Current RA treatments are largely empirical in origin and their precise mechanism of action is uncertain . Increasing evidence shows that chronic inflammatory diseases such as RA are caused by prolonged production of proinflammatory cytokines including tumor necrosis factor ( P01375 ) and interleukin 1 ( IL-1 ) . The nuclear factor kappaB ( NF-kappaB ) plays an essential role in transcriptional activation of P01375 and IL-1 . NF-kappaB is induced by many stimuli including P01375 and IL-1 , forming a positive regulatory cycle that may amplify and maintain RA disease process . NF-kappaB and enzymes involved in its activation can be a target for anti-inflammatory treatment . DB00945 and sodium salicylate inhibit activation of NF-KB by blocking O15111 , a key enzyme in NF-kappaB activation . Glucocorticoids suppress expression of inflammatory genes by binding glucocorticoid receptor with NF-kappaB , and increasing expression of inhibitory protein of NF-kappaB , P25963 . Sulfasalazine and gold compounds also inhibit NF-kappaB activation . Continuing advances in our understanding of action mechanism of antirheumatic agents will benefit the future development of RA regimens with greater efficacy and less toxicity . 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 . Thalidomide suppresses Up-regulation of human immunodeficiency virus coreceptors P61073 and P51681 on P01730 + T cells in humans . Concurrent infection in patients with human immunodeficiency virus ( HIV ) infection increases the expression of HIV coreceptors P61073 and P51681 . Thalidomide has beneficial effects in a number of HIV-associated diseases . The effect of thalidomide on P61073 and P51681 expression on P01730 + T cells was determined . Thalidomide produced a dose-dependent inhibition of lipopolysaccharide ( LPS ) -induced up-regulation of P61073 and P51681 in vitro . Antibody to tumor necrosis factor-alpha ( P01375 ) also attenuated the LPS-induced HIV coreceptor up-regulation , which was not further reduced by thalidomide . Thalidomide ( 400 mg ) was orally administered to 6 men , and their blood was stimulated ex vivo with LPS , staphylococcal or mycobacterial antigens , or antibody to CD3 or P10747 cells . All stimuli induced up-regulation of HIV coreceptors , which was reduced after ingestion of thalidomide . Thalidomide may be beneficial in the treatment of intercurrent infections during HIV infection by reducing the up-regulation of P61073 and P51681 expression on P01730 + T cells induced by bacterial and mycobacterial antigens , by a mechanism that involves inhibition of P01375 . DB00945 use may change cost-effectiveness of P35354 inhibitors . DB00945 modulates innate inflammatory response and inhibits the entry of Trypanosoma cruzi in mouse peritoneal macrophages . The intracellular protozoan parasite Trypanosoma cruzi causes Chagas disease , a serious disorder that affects millions of people in Latin America . Cell invasion by T. cruzi and its intracellular replication are essential to the parasite 's life cycle and for the development of Chagas disease . Here , we present evidence suggesting the involvement of the host 's cyclooxygenase ( P36551 ) enzyme during T. cruzi invasion . Pharmacological antagonist for P23219 , aspirin ( ASA ) , caused marked inhibition of T. cruzi infection when peritoneal macrophages were pretreated with ASA for 30 min at 37°C before inoculation . This inhibition was associated with increased production of IL-1β and nitric oxide ( NO(∙) ) by macrophages . The treatment of macrophages with either NOS inhibitors or prostaglandin E2 ( DB00917 ) restored the invasive action of T. cruzi in macrophages previously treated with ASA . Lipoxin Q96JZ2 -receptor antagonist Boc2 reversed the inhibitory effect of ASA on trypomastigote invasion . Our results indicate that DB00917 , NO(∙) , and lipoxins are involved in the regulation of anti-T. cruzi activity by macrophages , providing a better understanding of the role of prostaglandins in innate inflammatory response to T. cruzi infection as well as adding a new perspective to specific immune interventions . Update on aspirin desensitization for chronic rhinosinusitis with polyps in aspirin-exacerbated respiratory disease ( AERD ) . DB00945 -exacerbated respiratory disease ( AERD ) is a clinical condition which results in adverse upper and lower respiratory symptoms , particularly rhinitis , conjunctivitis , bronchospasm , and/or laryngospasm , following exposure to cyclooxygenase-1 ( P23219 ) inhibiting drugs , namely aspirin or nonsteroidal anti-inflammatory drugs ( NSAIDs ) . A provocative aspirin challenge is the gold standard for diagnosis of AERD . DB00945 desensitization and continuous aspirin therapy has been highly efficacious in those patients with suboptimal control of their disease on current available pharmacotherapy or those with other underlying conditions ( i.e. , cardiovascular disease ) who may require frequent treatment with aspirin or NSAIDs . This review article focuses on aspirin desensitization and the management of patients with AERD with a particular emphasis on outcomes in those patients with chronic rhinosinusitis and nasal polyposis . Osteoclast size heterogeneity in rat long bones is associated with differences in adhesive ligand specificity . P00734 ( PT ) is an RGD-containing bone-residing precursor to the serine protease thrombin ( TH ) , which acts as an agonist for a variety of cellular responses in osteoblasts and osteoclasts . We show here that PT , TH , osteopontin ( P10451 ) and fibronectin ( FN ) promoted adhesion of isolated neonatal rat long bone osteoclasts . However , the cells that adhered to PT and TH were smaller in size , rounded and contained 3-4 nuclei , in comparison to the cells adhering to P10451 and FN , which were larger with extended cytoplasmic processes and 6-7 nuclei . Attachment of the larger osteoclasts to P10451 and FN was inhibited by antibodies towards beta 3 and beta 1 integrin subunits , respectively . Whereas an RGD-containing peptide inhibited adhesion of the smaller osteoclasts to PT and TH , this was not seen with the beta 3 or beta 1 antibodies . In contrast , the beta 1 antibody augmented osteoclast adhesion to PT and TH in an RGD-dependent manner . Small osteoclasts were less efficient in resorbing mineralized bovine bone slices , as well as expressed lower mRNA levels of P14780 and the cathepsins K and L compared to large osteoclasts . The small osteoclast adhering to PT and TH may represent either an immature , less functional precursor to the large osteoclast or alternatively constitute a distinct osteoclast population with a specific role in bone . 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 . 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 . 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 . Role of Tyr348 in Tyr385 radical dynamics and cyclooxygenase inhibitor interactions in prostaglandin H synthase-2 . Both prostaglandin H synthase ( PGHS ) isoforms utilize a radical at Tyr385 to abstract a hydrogen atom from arachidonic acid , initializing prostaglandin synthesis . A Tyr348-Tyr385 hydrogen bond appears to be conserved in both isoforms ; this hydrogen bonding has the potential to modulate the positioning and reactivity of the Tyr385 side chain . The EPR signal from the Tyr385 radical undergoes a time-dependent transition from a wide doublet to a wide singlet species in both isoforms . In P35354 , this transition results from radical migration from Tyr385 to Tyr504 . Localization of the radical to Tyr385 in the recombinant human P35354 Y504F mutant was exploited in examining the effects of blocking Tyr385 hydrogen bonding by introduction of a further Y348F mutation . Cyclooxygenase and peroxidase activities were found to be maintained in the Y348F/Y504F mutant , but the Tyr385 radical was formed more slowly and had greater rotational freedom , as evidenced by observation of a transition from an initial wide doublet species to a narrow singlet species , a transition not seen in the parent Y504F mutant . The effect of disrupting Tyr385 hydrogen bonding on the cyclooxygenase active site structure was probed by examination of cyclooxygenase inhibitor kinetics . DB00945 treatment eliminated all oxygenase activity in the Y348F/Y504F double mutant , with no indication of the lipoxygenase activity observed in aspirin-treated wild-type P35354 . Introduction of the Y348F mutation also strengthened the time-dependent inhibitory action of nimesulide . These results suggest that removal of Tyr348-Tyr385 hydrogen bonding in P35354 allows greater conformational flexibility in the cyclooxygenase active site , resulting in altered interactions with inhibitors and altered Tyr385 radical behavior . P35354 induction and prostaglandin E2 accumulation in squamous cell carcinoma as a consequence of epidermal growth factor receptor activation by imatinib mesylate . Imatinib mesylate is a novel anti-tumor agent useful in the clinical management of chronic myelogenous leukemia and gastrointestinal stromal tumors with minimal toxicity relative to other forms of cancer therapy . Its clinical activity and minimal toxicity are related to specific inhibition of cellular targets including P11274 - P00519 , platelet-derived growth factor receptor and c-kit kinases , resulting in the collapse of downstream signaling cascades important for transformation . In some patients , unexpected toxicities arise that are not associated with inhibition of any known cellular imatinib target . In this report , we investigated the effects of imatinib on squamous carcinoma cell signaling . Imatinib induced expression of P35354 in a dose-dependent manner with concomitant accumulation of prostaglandin E2 . P35354 induction by imatinib was initiated through epidermal growth factor ( P01133 ) receptor kinase activation and downstream signaling through mitogenic-activated protein kinase . P35354 induction by imatinib was blocked by Q02750 or P01133 receptor inhibition . Imatinib did not activate stressor cytokine-signaling pathways ( p38 kinase , nuclear factor-kB nuclear translocation ) or affect P23219 expression . Imatinib failed to activate P01133 receptor signals in other tumor types , suggesting that P35354 induction in imatinib-treated cells is mediated through release of autocrine factors expressed or activated in squamous tumors . P35354 induction by imatinib in squamous tumors derived from the head and neck region is unique with respect to other target-specific agents and may represent one of the unintended toxic effects of imatinib described in some patients . Chromosomal changes in high- and low-invasive mouse lung adenocarcinoma cell strains derived from early passage mouse lung adenocarcinoma cell strains . The incidence of adenocarcinoma of the lung is increasing in the United States , however , the difficulties in obtaining lung cancer families and representative samples of early to late stages of the disease have lead to the study of mouse models for lung cancer . We used Spectral Karyotyping ( Q06418 ) , mapping with fluorescently labeled genomic clones ( Q5TCZ1 ) , comparative genomic hybridization ( CGH ) arrays , gene expression arrays , Western immunoblot and real time polymerase chain reaction ( PCR ) to analyze nine pairs of high-invasive and low-invasive tumor cell strains derived from early passage mouse lung adenocarcinoma cells to detect molecular changes associated with tumor invasion . The duplication of chromosomes 1 and 15 and deletion of chromosome 8 were significantly associated with a high-invasive phenotype . The duplication of chromosome 1 at band C4 and E1/2-H1 were the most significant chromosomal changes in the high-invasive cell strains . Mapping with Q5TCZ1 and CGH array further narrowed the minimum region of duplication of chromosome 1 to 71-82 centimorgans ( cM ) . Expression array analysis and confirmation by real time PCR demonstrated increased expression of P35354 , Q15631 ( TB- P02753 ) , O43781 , Q9H1E3 and Tubulin-alpha4 genes in the high-invasive cell strains . Elevated expression and copy number of these genes , which are involved in inflammation , cell movement , proliferation , inhibition of apoptosis and telomere elongation , were associated with an invasive phenotype . Similar linkage groups are altered in invasive human lung adenocarcinoma , implying that the mouse is a valid genetic model for the study of the progression of human lung adenocarcinoma . Tumor cell-derived prostaglandin E2 inhibits monocyte function by interfering with P51681 and Mac-1 . The cyclooxygenases ( P36551 ) -1 and P35354 are key enzymes in the conversion of arachidonic acid to prostaglandins and other eicosanoids . Whereas P23219 is expressed ubiquitously , P35354 is an immediate-early gene often associated with malignant transformation , and a role for the P36551 enzymes in tumor initiation and promotion is discussed . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) like aspirin and indomethacin that block P23219 and -2 have been shown to have beneficial effects for tumor patients . Therefore , these compounds have gained interest also among oncologists . However , the molecular mechanism by which NSAIDs inhibit carcinogenesis is not clearly understood . The prostaglandin-dependent and -independent effect may both account for their antineoplastic action . We show here that tumor cells derived from different tumors regularly produce prostaglandin E(2) ( PGE(2) ) interfering with the function of monocytes . In particular , PGE(2) inhibits the potential of monocytes to migrate in the direction of a chemotactic stimulus and to adhere to endothelial cell . This inhibition is most probably due to a modulation of the chemokine receptor P51681 and the beta2-integrin Mac-1 . Both down-regulation of P51681 and reduced expression of Mac-1 may diminish the potential of peripheral blood monocytes to leave blood vessels and invade target tissues . Since both dysfunctions can be restored with NSAIDs , our findings help to explain the molecular chemopreventive action of NSAIDs on tumor formation and progression . Senescence-associated superoxide dismutase influences mitochondrial gene expression in budding tunicates . A recent study has shown that in the budding tunicate Polyandrocarpa misakiensis , the mitochondrial respiratory chain ( MRC ) dramatically attenuates the gene activity during senescence . In this study , we examined the possible involvement of superoxide dismutase ( SOD ) in the attenuation of gene expression of cytochrome c oxidase subunit 1 ( P23219 ) in aged zooids . By RT-PCR and in situ hybridization , Cu/Zn-SOD ( P00441 ) was found to be expressed in most cells and tissues of buds and juvenile zooids but showed a conspicuous decline in senescent adult zooids , except in the gonad tissue in which the cytoplasm of juvenile oocytes was stained heavily . This expression pattern of P00441 was similar to that of P23219 . In contrast to P00441 , Mn-SOD ( P04179 ) was expressed constitutively in both somatic and germline tissues of buds , juvenile zooids , and senescent adult zooids . Knockdown of P00441 by RNAi diminished the gene activity of not only P00441 but also of P23219 . The resultant zooids had transient deficiencies in growth and budding , and they recovered from these deficiencies approximately 1 month later . Our results indicate that in P. misakiensis , P00441 is a senescence-associated nuclear gene and that the experimental decline in P00441 gene expression accompanies the attenuation of MRC gene activity . Although it is uncertain how P00441 is downregulated during tunicate senescence , the decreased P00441 activity could be one of the main causes of MRC gene attenuation during normal senescence . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . A computational prospect to aspirin side effects : aspirin and P23219 interaction analysis based on non-synonymous SNPs . DB00945 ( ASA ) is a commonly used nonsteroidal anti-inflammatory drug ( NSAID ) , which exerts its therapeutic effects through inhibition of cyclooxygenase ( P36551 ) isoform 2 ( P35354 ) , while the inhibition of P23219 by ASA leads to apparent side effects . In the present study , the relationship between P23219 non-synonymous single nucleotide polymorphisms ( nsSNPs ) and aspirin related side effects was investigated . The functional impacts of 37 nsSNPs on aspirin inhibition potency of P23219 with P23219 /aspirin molecular docking were computationally analyzed , and each SNP was scored based on DOCK Amber score . The data predicted that 22 nsSNPs could reduce P23219 inhibition , while 15 nsSNPs showed increasing inhibition level in comparison to the regular P23219 protein . In order to perform a comparing state , the Amber scores for two Arg119 mutants ( R119A and R119Q ) were also calculated . Moreover , among nsSNP variants , rs117122585 represented the closest Amber score to R119A mutant . A separate docking computation validated the score and represented a new binding position for ASA that acetyl group was located within the distance of 3.86Å from Ser529 OH group . This could predict an associated loss of activity of ASA through this nsSNP variant . Our data represent a computational sub-population pattern for aspirin P23219 related side effects , and provide basis for further research on P23219 /ASA interaction . 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 . 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 . DB00945 -like molecules that covalently inactivate cyclooxygenase-2 . Many of aspirin 's therapeutic effects arise from its acetylation of cyclooxygenase-2 ( P35354 ) , whereas its antithrombotic and ulcerogenic effects result from its acetylation of P23219 . Here , aspirin-like molecules were designed that preferentially acetylate and irreversibly inactivate P35354 . The most potent of these compounds was o-(acetoxyphenyl)hept-2-ynyl sulfide ( APHS ) . Relative to aspirin , APHS was 60 times as reactive against P35354 and 100 times as selective for its inhibition ; it also inhibited P35354 in cultured macrophages and colon cancer cells and in the rat air pouch in vivo . Such compounds may lead to the development of aspirin-like drugs for the treatment or prevention of immunological and proliferative diseases without gastrointestinal or hematologic side 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 . 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 . DB00945 and Other P23219 inhibitors . Currently available antiplatelet drugs interfere with the process of platelet activation and aggregation by selectively blocking key enzymes involved in the synthesis of platelet agonists , or membrane receptors mediating activation signals . Pharmacological interference with critical molecular pathways of platelet activation and aggregation may reduce the risk of atherothrombotic complications through mechanisms that are also responsible for an increased risk of bleeding . Acetylsalicylic acid ( aspirin ) represents a prototypic antiplatelet agent . The aim of this chapter is to integrate our current understanding of the molecular mechanism of action of aspirin with the results of clinical trials and epidemiological studies assessing its efficacy and safety . Moreover , the antiplatelet properties of reversible inhibitors of the same drug target will also be reviewed . 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 . Physiological mediators in nonsteroidal anti-inflammatory drugs ( NSAIDs ) -induced impairment of gastric mucosal defense and adaptation . Focus on nitric oxide and lipoxins . Prostaglandins mediate various physiological aspects of mucosal defense and the suppression of prostaglandin synthesis in the stomach is a critical event in terms of the development of mucosal injury after NSAID administration . However , it has become clear that other mediators besides prostaglandins can similarly act to protect the stomach from injury . For instance , nitric oxide ( NO ) released from vascular epithelium , epithelial cells of gastrointestinal tract and sensory nerves can influence many of the same components of mucosal defense as do prostaglandins . Thus , administration of NO in a form of NO-donors exert protective influence on the stomach from the injury that usually occurs when mucosal prostaglandin levels are suppressed . The new class of NO releasing NSAIDs , including NO-aspirin , represent a very promising approach to reducing the toxicity of anti-inflammatory drugs . Lipoxins are another group of lipid mediators that can protect the stomach . DB00945 -triggered lipoxin synthesis , via P35354 , acts to reduce the severity of damage induced by this drug . Lipoxin analogues may prove to be useful for preventing mucosal injury and for modulating mucosal inflammation . DB00945 -triggered lipoxin also seems to play in important role in gastric adaptation during chronic aspirin administration . Suppression of P35354 activity by selective P35354 inhibitors abolishes the production of this endogenous gastroprotective substance and diminishes the gastric tolerability of NSAIDS and gastric adaptation to these drugs . This review was designed to give an updated overview on the physiological factors and experimental and clinical attempts that were used or may be used in the future as the therapeutic approach to counteract adverse effects in the stomach associated with NSAID ingestion . DB00945 triggers formation of anti-inflammatory mediators : New mechanism for an old drug . Extract : DB00945 is a widely used non-steroidal anti-inflammatory drug ( NSAID ) . It is well documented that aspirin irreversibly inhibits cyclooxygenase ( P36551 ) by acetylation of an amino acid serine residue , and thus blocks the subsequent biosynthesis of prostaglandins and thromboxane . P36551 has at least two forms , P23219 and P35354 . P23219 is the main form present in mature platelets in the blood , where it transforms arachidonic acid to the intermediates PG-G/H , which are subsequently converted to thromboxane A2 . Thromboxane A2 is a vasoconstrictor and potent platelet activator . Thus , inhibition of thromboxane A2 formation explains aspirin 's anti-thrombotic properties . In the early 1990s , a second form of P36551 was identified , namely P35354 . P35354 was initially conceptualized as an " inducible " P36551 that is elevated in its quantity by a wide range of agents that stimulate inflammation or cell division and seems to be responsible for local formation during inflammation and cancer . 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 . DB00945 treatment influences platelet-related inflammatory biomarkers in healthy individuals but not in acute stroke patients . OBJECTIVES : Platelet-leukocyte aggregation is believed to contribute to acute thrombotic events . While the effect of aspirin on platelet-to-platelet aggregation is well established , the impact of the drug on pro-inflammatory platelet function remains equivocal . Thus we investigated the effect of aspirin on selected platelet-related inflammatory biomarkers in both acute ischaemic stroke patients and healthy volunteers . METHODS : Using five-colour flow cytometry the platelet surface expression of CD62P and P29965 and subpopulations of leukocyte-platelet aggregates were assessed in 63 acute stroke patients and 40 healthy volunteers at baseline and after a 10-day period of aspirin intake at a daily dose of 150 mg . Simultaneously the plasma levels of soluble CD62P and P29965 , serum level of TxB(2) , and whole blood impedance platelet aggregation under arachidonic acid ( AA ) stimulation were investigated . RESULTS : No differences in values of studied platelet-related inflammatory biomarkers in both resting platelets and those activated with TRAP after 10-day treatment with aspirin were confirmed in stroke subjects . In healthy individuals the resting platelet expression of CD62P , plasma level of soluble CD62P and percentage of circulating monocyte-platelet aggregates were lower after the aspirin intake period ( P=0.009 ; P=0.04 ; P=0.004 , respectively ) . In both studied groups serum level of TxB(2) and platelet aggregation under AA stimulation were lower than before treatment ( P < 0.001 ) . CONCLUSION : Despite effective inhibition of P23219 -dependent platelet aggregation , aspirin does not influence the platelet α-granule-derived inflammatory mediators and monocyte-platelet aggregation in acute stroke subjects , although it does in healthy individuals . DB00945 resistance . DB00945 resistance is the inability of aspirin to reduce platelet production of thromboxane A2 and thereby platelet activation and aggregation . Increasing degrees of aspirin resistance may correlate independently with increasing risk of cardiovascular events . DB00945 resistance can be detected by laboratory tests of platelet thromboxane A2 production or platelet function that depend on platelet thromboxane production . Potential causes of aspirin resistance include inadequate dose , drug interactions , genetic polymorphisms of P23219 and other genes involved in thromboxane biosynthesis , upregulation of non-platelet sources of thromboxane biosynthesis , and increased platelet turnover . DB00945 resistance can be overcome by treating the cause or causes , and reduced by minimising thromboxane production and activity , and blocking other pathways of platelet activation . Future research is aimed at defining aspirin resistance , developing reliable tests for it , and establishing the risk of associated cardiovascular events . Potential mechanisms of aspirin resistance can then be explored and treatments assessed . 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 . DB00945 and non-small cell lung cancer resections : effect on long-term survival . OBJECTIVE : Survival after resections for non-small cell lung cancer remains poor . Recurrent lung cancer remains common . Due to the common risk factor of smoking , cardiovascular deaths occur in the absence of recurrent lung cancer in up to 15 % of patients . DB00945 has been proven to reduce cardiovascular mortality as a secondary prophylactic agent , but not as a primary agent . DB00945 being a P35354 inhibitor has been shown to reduce the chance of metastasis in adenocarcinoma but not squamous carcinoma . We sought to investigate the effect of long-term aspirin therapy on survival post potentially curative surgery . METHODS : We analysed a prospective thoracic surgical database , from time period 2003 to date . Patients who were on aspirin pre-operatively , N=412 were compared to non users , N=1353 . Patient long-term outcome was assessed utilising the national strategic tracking service that operates in the United Kingdom . Cox proportional hazards analysis was used to determine significant factors affecting survival . RESULTS : 100 % survival follow up was achieved . Regular users of aspirin had > 5 % increased survival , which was significant , p=0.05 , despite having a higher cardiovascular risk profile . Mode of death data was not available . CONCLUSIONS : Adjuvant aspirin post resection for potentially curative non-small cell lung cancer significantly increases survival . The mechanism of increased survival needs further investigation and is the basis for the trial : Adjuvant DB00945 for Non-Small cell Lung Cancer -- The Big A Trial. www.TheBigATrial.co.uk . [ DB00945 -- the prodigious panacea ? Molecular mechanisms of the action of acetylsalicylic acid in the organism ] . DB00945 ( acetylsalicylic acid ) is a commonly used non-steroidal anti-inflammatory drug capable of acetylating proteins in the course of a simple , non-enzymatic chemical reaction . Its main physiological effect is inhibiting prostanoid synthesis . Cyclooxygenases , P23219 and P35354 , are crucial in the metabolic pathway leading to the generation of prostanoids . Both enzymes are major cellular targets for aspirin . The physiological spectrum of the biological activity of the prostanoids is very broad , and underlies the high clinical effectiveness of aspirin as an anti-inflammatory , antipyretic , and analgesic drug . Apart from the inhibition of prostanoid synthesis aspirin shows a variety of pharmacological activities , including reduction of DB00171 storage pools , increased extracellular adenosine , lowered inducible nitric oxide synthase activity , modulation of mitogen-activated protein kinases , and the expression of a plethora of genes induced under conditions of cell stress via the regulation of transcription factor NFkappaB activity . Such multipotent action explains its wide use in clinical practice . Regardless of the accumulated evidence on the molecular mechanisms of aspirin 's action , the rationale of the appropriate dosing and monitoring of aspirin therapy and prophylaxis remains obscure . Hence , an evaluation and reasonable weighing of the cost/benefit ratio of aspirin therapy in various diseases seems appropriate . 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 . 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 . [ DB00945 and prevention of colorectal carcinomas ] . Observational and some randomized clinical trials suggest that aspirin protects from occurrence and progression of colorectal neoplasias ( adenomas , carcinomas ) . However , there are still open questions , regarding the benefit/risk ratio ( bleedings ) as well as dosage and duration of treatment during the probably long-term medication , before stringent recommendations regarding clinical use of aspirin can be made . Specifically , there is currently no generally accepted mode of action or molecular target of aspirin , though a relationship to tumor-associated enhanced DB00917 levels in the affected mucosa is likely . Regular daily intake of aspirin in antiplatelet doses of 100 mg appears to be sufficient in responding persons . If this is confirmed in prospective randomized trials that are currently underway , this might add to the prophylactic use of aspirin and would suggest a pharmacological relationship to inhibition of P23219 mediated prostaglandin/thromboxane biosynthesis as a common primary target for both cardiocoronary and antineoplastic prophylaxis . Prophylactic aspirin use might then add to an undoubtedly important healthy lifestyle including appropriate diet . DB00945 inhibits TNFalpha- and IL-1-induced NF-kappaB activation and sensitizes HeLa cells to apoptosis . Rel/nuclear factor-kappa B ( NF-kappaB ) transcription factors are involved in transcription of several target genes that modulate proliferation , apoptosis and cell growth . TNFalpha- and IL-1-induced NF-kappaB activation pathways mainly involve the phosphorylation and degradation of P25963 by a signalsome complex followed by nuclear translocation of NF-kappaB and target gene expression . NF-kappaB mediates the balance between cell death and survival as most cancer cells that have rather constitutive or inducible activation of NF-kappaB are resistant to apoptosis even by strong apoptotic agents such as TNFalpha . In this study we demonstrate that proinflammatory cytokines TNFalpha and IL-1 induced NF-kappaB activation in human cervical carcinoma HeLa cells . Our studies reveal that acetylsalicylic acid ( aspirin ) prevents TNFalpha- and IL-1-induced NF-kappaB activation in a dose-dependent manner through inhibition of phosphorylation and degradation of P25963 and Q15653 . Moreover , aspirin sensitizes HeLa cells to TNFalpha-induced apoptosis . These results suggest that aspirin could be used to potentiate the effectiveness of TNFalpha-based therapeutic interventions in cancer treatment . 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 . 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 . 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 . 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 .

MH_train_1133

MH_train_1133

MH_train_1133

interacts_with DB09036?

multiple_choice

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 . 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 . 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 . 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 . 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 . Combined effects of C225 and 125-iodine seed radiation on colorectal cancer cells . BACKGROUND : To characterize the effect of combined treatment of the anti-epidermal growth factor receptor ( P00533 ) monoclonal antibody C225 and 125-iodine ( 125I ) seed radiation in human colorectal cancer . METHODS : We treated LS180 cells with 125I continuous low dose rate radiation in the presence and absence of 100 nM C225 . The clonogenic capacity , cellular proliferation , cell cycle distribution , apoptosis , and molecular pathways of the cells following the treatments were analyzed in vitro . RESULTS : The sensitizer enhancement ratio of C225 was approximately 1.4 . Treatment with C225 and radiation alone produced significant inhibition of cell growth , but combination therapy produced greater inhibition than either treatment administered alone . C225 increased the radiation-induced apoptosis and the fraction of γ- P16104 foci positive cells at 48 h after treatment . The Akt phosphorylation level was lower in the cells receiving the combination treatment than in the cells treated with radiation or C225 alone . CONCLUSIONS : These findings indicate that C225 sensitizes LS180 cells to 125I seed radiation . Growth inhibition is mediated by inducing apoptosis and not cell cycle arrest . Additionally , we confirmed that C225 impairs DNA repair by reducing the cellular level of the P78527 and P12956 proteins . Furthermore , the inhibition of Akt signaling activation may be responsible for the C225-mediated radiosensitization . 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 . 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 . 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 . 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 . Involvement of Rho-kinase in tumor necrosis factor-alpha-induced interleukin-6 release from P13671 glioma cells . P01375 ( P01375 ) -alpha stimulated interleukin ( IL ) -6 release and induced the phosphorylation of myosin phosphatase targeting subunit ( MYPT ) -1 , a Rho-kinase substrate . The P05231 release was significantly suppressed by Y-27632 and fasudil , Rho-kinase inhibitors . Although IkappaB inhibitor suppressed the P01375 -induced P05231 release , the Rho-kinase inhibitors did not affect the P01375 -induced IkappaB phosphorylation . P01375 induced the phosphorylation of p38 mitogen-activated protein ( Q96HU1 ) kinase , stress-activated protein kinase ( SAPK ) /c-Jun N-terminal kinase ( JNK ) , and Q8TCB0 / Q8NFH3 Q96HU1 kinase . The P01375 -induced P05231 release was suppressed by SB203580 , a p38 MAPK inhibitor , or SP600125 , a SAPK/JNK inhibitor , but not by PD98059 , a Q96HU1 kinase/extracellular signal-regulated kinase kinase inhibitor . The Rho-kinase inhibitors attenuated the P01375 -induced phosphorylation of both p38 Q96HU1 kinase and SAPK/JNK . Rho-kinase , which has been used for the clinical treatment of cerebral vasospasms , may be involved in other central nervous system ( CNS ) disorders such as traumatic injury , stroke , neurodegenerative disease and neuropathic pain . P01375 , a proinflammatory cytokine that affects the CNS through cytokines , such as P05231 , release from neurons , astrocytes and microglia . Therefore , we investigated the involvement of Rho-kinase in the P01375 -induced P05231 release from rat P13671 glioma cells . These results strongly suggest that Rho-kinase regulates the P01375 -induced P05231 release at a point upstream from p38 MAPK and SAPK/JNK in P13671 glioma cells . Therefore , Rho-kinase inhibitor may be considered to be a new clinical candidate for the treatment of CNS disorders in addition to cerebral vasospasms . Tanshinone IIA inhibits constitutive P40763 activation , suppresses proliferation , and induces apoptosis in rat P13671 glioma cells . P40763 ( P40763 ) is usually constitutively activated in a variety of malignancies . Thus , P40763 may be a promising target for treatment of tumor cells . Recently , Tanshinone IIA ( Tan IIA ) , a major active constituent from the root of Salvia miltiorrhiza Bunge , was reported to have apoptosis inducing effects on a large variety of cancer cells . In this study , we evaluate the anti-proliferation and apoptosis inducing effects of Tan IIA on P13671 glioma cells . Cell growth and proliferation were measured by MTT assay , cell apoptosis was observed by flow cytometry and DNA-fragmentation analysis . Further more , we investigated inhibitory effects of Tan IIA on P40763 activity and its downstream targets : Bcl-XL , cyclin D1 . Alteration of P40763 activity was examined by measuring their DNA binding activity and tyrosine phosphorylation . Changes in the expression levels of Bcl-XL and cyclin D1 were examined by Western blot analysis . We found that the cellular growth were inhibited and cell apoptosis were observed after the treatment with Tan IIA . The P40763 activity was significantly reduced by Tan IIA parallel with a significant attenuation of expression of Bcl-XL and cyclin D1 . These results suggest that Tan IIA may serve as an effective adjunctive reagent in the treatment of glioma for its targeting of constitutive P40763 signaling . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 ) .

MH_train_1134

MH_train_1134

MH_train_1134

interacts_with DB06603?

multiple_choice

Expression of Npc1 in glial cells corrects sterility in Npc1(-/-) mice . Niemann-Pick type C1 ( NPC ) disease is an autosomal recessive neurodegenerative disorder . One feature of the mouse model of O15118 is it 's infertility . We have made transgenic mice which express the Npc1 protein exclusively in fibrillary astrocytes , using the glial fibrillary acidic protein ( P14136 ) promoter . This selective expression of Npc1 corrects sterility in P14136 -Npc1(-/-) , Npc1(-/-) mice . Counts of acidophils in the pituitary of P14136 -Npc1E , Npc1(-/-) mice , as compared Npc1(-/-) mice , and measurements of dopamine D2 receptor ( P14416 ) mRNA in the pituitary , suggest mechanisms for fertility enhancement . We conclude that the correction of sterility in P14136 -Npc1E , Npc1(-/-) mice is a result of restoring hypothalamic control of the pituitary . 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 . 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 . Class I histone deacetylase-mediated repression of the proximal promoter of the activity-regulated cytoskeleton-associated protein gene regulates its response to brain-derived neurotrophic factor . We examined the transcriptional regulation of the activity-regulated cytoskeleton-associated protein gene ( Arc ) , focusing on P23560 -induced Arc expression in cultured rat cortical cells . Although the synaptic activity-responsive element ( SARE ) , located -7 kbp upstream of the Arc transcription start site , responded to DB01221 , P23560 , or P09038 , the proximal region of the promoter ( Arc/-1679 ) was activated by P23560 or P09038 , but not by DB01221 , suggesting the presence of at least two distinct Arc promoter regions , distal and proximal , that respond to extracellular stimuli . Specificity protein 4 ( Q02446 ) and early growth response 1 ( P18146 ) controlled Arc/-1679 transcriptional activity via the region encompassing -169 to -37 of the Arc promoter . We found that trichostatin A ( P32119 ) , a histone deacetylase ( HDAC ) inhibitor , significantly enhanced the inductive effects of P23560 or P09038 , but not those of DB01221 on Arc expression . Inhibitors of class I/IIb HDACs , DB02546 , and class I HDACs , MS-275 , but not of class II HDACs , MC1568 , enhanced P23560 -induced Arc expression . The enhancing effect of P32119 was mediated by the region from -1027 to -1000 bp , to which serum response factor ( P11831 ) and Q13547 bound . The binding of Q13547 to this region was reduced by P32119 . Thus , Arc expression was suppressed by class I HDAC-mediated mechanisms via chromatin modification of the proximal promoter whereas the inhibition of HDAC allowed Arc expression to be markedly enhanced in response to P23560 or P09038 . These results contribute to our understanding of the physiological role of Arc expression in neuronal functions such as memory consolidation . 9-Hydroxystearic acid interferes with P01133 signalling in a human colon adenocarcinoma . The epidermal growth factor has long been known to be strictly correlated with the highly proliferating activities of cancer cells and primary tumors . Moreover , in the nucleus , the epidermal growth factor/epidermal growth factor receptor complex ( P01133 / P00533 ) functions as a transcriptional regulator that activates the cyclin D1 gene . 9-hydroxystearic acid ( 9-HSA ) induces cell proliferation arrest and differentiation in HT29 colon cancer cells by inhibiting histone deacetylase 1 ( Q13547 ) . 9-HSA-treated HT29 , when stimulated with P01133 , are not responsive and surprisingly undergo a further arrest . In order to understand the mechanisms of this effect , we analyzed the degree of internalization of the P01133 / P00533 complex and its interactions with Q13547 . It appears that Q13547 , as modified by 9-HSA , is unable to associate with cyclin D1 , interfering with the cell proliferation program , and sequesters the P01133 / P00533 complex interrupting the transduction of the mitogenic signal . Clinical characterization of individuals with deletions of genes in holoprosencephaly pathways by aCGH refines the phenotypic spectrum of HPE . Holoprosencephaly ( HPE ) is the most common developmental forebrain anomaly in humans . Both environmental and genetic factors have been identified to play a role in the HPE phenotype . Previous studies of the genetic bases of HPE have taken a phenotype-first approach by examining groups of patients with HPE for specific mutations or deletions in known or candidate HPE genes . In this study , we characterized the presence or absence of HPE or a microform in 136 individuals in which microarray-based comparative genomic hybridization ( aCGH ) identified a deletion of one of 35 HPE loci . Frank holoprosencephaly was present in 11 individuals with deletions of one of the common HPE genes SHH , O95409 , O95343 , and Q15583 , in one individual with a deletion of the HPE8 locus at 14q13 , and in one individual with a deletion of P55075 , whereas deletions of other HPE loci and candidate genes ( Q9Y261 and P98164 ) expressed microforms of HPE . Although individuals with deletions of other HPE candidates ( Q96F81 , P48449 , Q96QV1 , SMO , P12644 , Q4KMG0 , P60953 , P27037 , P32243 , and Q9Y5W5 ) had clinically significant features , none had frank HPE or a microform . A search for significant aCGH findings in individuals referred for testing for HPE revealed a novel association of a duplication involving P49841 at 3q13.33 with HPE or a microform , seen in two unrelated individuals . 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 . P10275 signaling : mechanism of interleukin-6 inhibition . Nonsteroidal signaling via the androgen receptor ( AR ) plays an im-portant role in hormone-refractory prostate cancer . Previously , we have reported that the pleiotropic cytokine , interleukin ( IL ) -6 , inhibited dihydrotestosterone-mediated expression of prostate-specific antigen in LNCaP cells ( Jia et al. , Mol Can Res 2003;1:385-92 ) . In the present study , we explored the mechanisms involved in this inhibition and considered possible effects on AR nuclear translocation , recruitment of transcription cofactors , and the signaling pathways that may mediate this inhibitory effect . P05231 neither induced nuclear localization of the AR nor inhibited dihydrotestosterone-induced nuclear translocation of the receptor . P05231 did not affect AR or P52701 coactivator recruitment to the transcription initiation complex on the prostate-specific antigen enhancer and promoter . Moreover , it did not lead to the recruitment of the corepressor silencing mediator of retinoic acid and thyroid hormone receptor ( Q9Y618 ) or histone deacetylase 1 ( Q13547 ) at the same sites . P05231 did , however , prevent the recruitment of the secondary coactivator , p300 , to the complex and partially inhibited histone H3 acetylation at the same loci . Furthermore , inhibition by P05231 was not mediated by the mitogen-activated protein kinase or the Akt pathways and was partially abrogated by signal transducers and activators of transcription-3 knock-down using small interfering RNA . Our results show that P05231 modulates androgen action through the differential recruitment of cofactors to target genes . These findings may account for the pleiotropic actions of P05231 in malignant prostate cells . 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 . [ 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 . 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 . In utero exposure to low doses of environmental pollutants disrupts fetal ovarian development in sheep . Epidemiological studies of the impact of environmental chemicals on reproductive health demonstrate consequences of exposure but establishing causative links requires animal models using ' real life ' in utero exposures . We aimed to determine whether prolonged , low-dose , exposure of pregnant sheep to a mixture of environmental chemicals affects fetal ovarian development . Exposure of treated ewes ( n = 7 ) to pollutants was maximized by surface application of processed sewage sludge to pasture . Control ewes ( n = 10 ) were reared on pasture treated with inorganic fertilizer . Ovaries and blood were collected from fetuses ( n = 15 control and n = 8 treated ) on Day 110 of gestation for investigation of fetal endocrinology , ovarian follicle/oocyte numbers and ovarian proteome . Treated fetuses were 14 % lighter than controls but fetal ovary weights were unchanged . P01236 ( 48 % lower ) was the only measured hormone significantly affected by treatment . Treatment reduced numbers of growth differentiation factor ( O60383 ) and induced myeloid leukaemia cell differentiation protein ( Q07820 ) positive oocytes by 25-26 % and increased pro-apoptotic Q07812 by 65 % and 42 % of protein spots in the treated ovarian proteome were differently expressed compared with controls . Nineteen spots were identified and included proteins involved in gene expression/transcription , protein synthesis , phosphorylation and receptor activity . Fetal exposure to environmental chemicals , via the mother , significantly perturbs fetal ovarian development . If such effects are replicated in humans , premature menopause could be an outcome . 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 . 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 . Characterization of human mesenchymal stem cell secretome at early steps of adipocyte and osteoblast differentiation . BACKGROUND : It is well established that adipose tissue plays a key role in energy storage and release but is also a secretory organ and a source of stem cells . Among different lineages , stem cells are able to differentiate into adipocytes and osteoblasts . As secreted proteins could regulate the balance between both lineages , we aimed at characterizing the secretome of human multipotent adipose-derived stem cell ( hMADS ) at an early step of commitment to adipocytes and osteoblasts . RESULTS : A proteomic approach , using mono-dimensional electrophoresis and tandem mass spectrometry , allowed us to identify a total of 73 proteins at day 0 and day 3 of adipocyte and osteoblast differentiation . Analysis of identified proteins showed that 52 % corresponded to classical secreted proteins characterized by a signal peptide , that 37 % previously described in the extracellular compartment were devoid of signal peptide and that 11 % neither exhibited a signal peptide nor had been previously described extracellularly . These proteins were classified into 8 clusters according to their function . Quantitative analysis has been performed for 8 candidates : P05121 , P36955 , Q15582 , PTX3 , P09486 , P06733 , P11021 and P08253 . Among them , P05121 was detected at day 0 and day 3 of osteoblast differentiation but never in adipocyte secretome . Furthermore we showed that P05121 mRNA was down-regulated in the bone of ovariectomized mice . CONCLUSION : Given its regulation during the early events of hMADS cell differentiation and its status in ovariectomized mice , P05121 could play a role in the adipocyte/osteoblast balance and thus in bone diseases such as osteoporosis . 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 . 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 . 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 . 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 . 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 . Fibroblast growth factor-2 in hyperplastic pituitaries of D2R knockout female mice . P14416 ( D2R ) knockout ( KO ) female mice develop chronic hyperprolactinemia and pituitary hyperplasia . Our objective was to study the expression of the mitogen fibroblast growth factor ( P09038 ) and its receptor , P11362 , comparatively in pituitaries from KO and wild-type ( WT ) female mice . We also evaluated P09038 subcellular localization and P09038 effects on pituitary function . P09038 -induced prolactin release showed a similar response pattern in both genotypes , even though basal and P09038 -stimulated release was higher in KO . P09038 stimulated pituitary cellular proliferation ( MTS assay and [(3)H]thymidine incorporation ) , with no differences between genotypes . P09038 concentration ( measured by ELISA ) in whole pituitaries or cultured cells was lower in KO ( P < 0.00001 and 0.00014 ) . Immunofluorescence histochemistry showed less P09038 in pituitaries from KO females and revealed a distinct P09038 localization pattern between genotypes , being predominantly nuclear in KO and cytosolic in WT pituitaries . Finally , P09038 could not be detected in the conditioned media from pituitary cultures of both genotypes . P11362 levels ( Western blot and immunohistochemistry ) were higher in pituitaries of KO . Basal concentration of phosphorylated ERKs was lower in KO cells ( P = 0.018 ) . However , when stimulated with P09038 , a significantly higher increment of P29323 phosphorylation was evidenced in KO cells ( P < or = 0.02 ) . We conclude that disruption of the D2R caused an overall decrease in pituitary P09038 levels , with an increased distribution in the nucleus , and increased P11362 levels . These results are important in the search for reliable prognostic indicators for patients with pituitary dopamine-resistant prolactinomas , which will make tumor-specific therapy possible . An ataxia telangiectasia model : inefficient cell differentiation and possible reversal by serine protease inhibitors , tumor necrosis factor inhibitors , dexamethasone , and glutathione enhancers . Ataxia telangiectasia ( AT ) is a rare genetic disorder . Symptoms of the disease include cerebellar ataxia , depressed immunoresponsiveness , increased sensitivity to radiation , and leukemias . Various kinds of AT cells show reduced efficiency of differentiation . The ataxia telangiectasia gene ( Q13315 ) may reduce differentiation by suppressing cell responsivity to insulin . P01308 sensitivity seems lower in AT . P01375 may overactivate NF-kappa B in AT , and this increases the radiosensitivity of AT cells . Intracellular reduced glutathione may also become depleted . The reduced levels of glutathione may further alter differentiation of AT cells . DB00133 protease inhibitors may counteract the effects of tumor necrosis factor . Glutathione enhancers may also prove valuable as therapy . 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 . 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 . Increasing P16070 +/ P25063 (-) tumor stem cells , and upregulation of P35354 and Q9UBN7 , as major functions of P04626 in breast tumorigenesis . BACKGROUND : Cancer cells are believed to arise primarily from stem cells . P16070 +/ P25063 (-) have been identified as markers for human breast cancer stem cells . Although , P04626 is a well known breast cancer oncogene , the mechanisms of action of this gene are not completely understood . Previously , we have derived immortal ( M13SV1 ) , weakly tumorigenic ( M13SV1R2 ) and highly tumorigenic ( M13SV1R2N1 ) cell lines from a breast epithelial cell type with stem cell phenotypes after successive SV40 large T-antigen transfection , X-ray irradiation and ectopic expression of P04626 /C-erbB2/neu . Recently , we found that M13SV1R2 cells became non-tumorigenic after growing in a growth factor/hormone-deprived medium ( R2d cells ) . RESULTS : In this study , we developed M13SV1R2N1 under the same growth factor/hormone-deprived condition ( R2N1d cells ) . This provides an opportunity to analyze P04626 effect on gene expression associated with tumorigenesis by comparative study of R2d and R2N1d cells with homogeneous genetic background except P04626 expression . The results reveal distinct characters of R2N1d cells that can be ascribed to P04626 : 1 ) development of fast-growing tumors ; 2 ) high frequency of P16070 +/ P25063 (-) cells ( ~50 % for R2N1d vs. ~10 % for R2d ) ; 3 ) enhanced expression of P35354 , Q9UBN7 mediated , respectively , by MAPK and PI3K/Akt pathways , and many genes associated with inflammation , metastasis , and angiogenesis . Furthermore , P04626 expression can be down regulated in non-adhering R2N1d cells . These cells showed longer latent period and lower rate of tumor development compared with adhering cells . CONCLUSIONS : P04626 may induce breast cancer by increasing the frequency of tumor stem cells and upregulating the expression of P35354 and Q9UBN7 that play pivotal roles in tumor progression . 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 . Effect of dietary NaCl on tyrosine hydroxylase in the superior cervical ganglia of Dahl rats . To investigate the involvement of peripheral catecholamines in the development of Dahl-Iwai salt-sensitive ( Q8IX12 /Eis ) hypertension , we performed immunohistochemical staining of tyrosine hydroxylase ( TH ) in the superior cervical ganglia ( SCG ) of Q8IX12 /Eis rats and Dahl-Iwai salt-resistant ( P30518 /Eis ) rats , and in situ hybridization histochemistry for demonstration of TH mRNA localization in the SCG of these rats . Q8IX12 /Eis and P30518 /Eis rats were fed on a high ( 8 % ) salt diet or on a low ( 0.3 % ) salt diet for 4 weeks . Nerve cells in the SCG of Q8IX12 /Eis high salt rats exhibited more intense TH-immunoreactivity ( P < 0.01 ) and hybridization signals ( P < 0.01 ) than those of the other experimental groups . These findings suggest that activation of peripheral sympathetic nerves may account for hypertension in Q8IX12 /Eis rats on a high salt diet . A transcriptional repressor co-regulatory network governing androgen response in prostate cancers . Transcriptional corepressors are frequently aberrantly over-expressed in prostate cancers . However , their crosstalk with the P10275 ( AR ) , a key player in prostate cancer development , is unclear . Using ChIP-Seq , we generated extensive global binding maps of AR , ERG , and commonly over-expressed transcriptional corepressors including Q13547 , Q92769 , O15379 , and Q15910 in prostate cancer cells . Surprisingly , our results revealed that ERG , HDACs , and Q15910 are directly involved in androgen-regulated transcription and wired into an AR centric transcriptional network via a spectrum of distal enhancers and/or proximal promoters . Moreover , we showed that similar to ERG , these corepressors function to mediate repression of AR-induced transcription including cytoskeletal genes that promote epithelial differentiation and inhibit metastasis . Specifically , we demonstrated that the direct suppression of P18206 expression by ERG , Q15910 , and HDACs leads to enhanced invasiveness of prostate cancer cells . Taken together , our results highlight a novel mechanism by which , ERG working together with oncogenic corepressors including HDACs and the polycomb protein , Q15910 , could impede epithelial differentiation and contribute to prostate cancer progression , through directly modulating the transcriptional output of AR . 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 . Identifying new targets in leukemogenesis using computational approaches . There is a need to identify novel targets in Acute Lymphoblastic Leukemia ( ALL ) , a hematopoietic cancer affecting children , to improve our understanding of disease biology and that can be used for developing new therapeutics . Hence , the aim of our study was to find new genes as targets using in silico studies ; for this we retrieved the top 10 % overexpressed genes from Oncomine public domain microarray expression database ; 530 overexpressed genes were short-listed from Oncomine database . Then , using prioritization tools such as ENDEAVOUR , P30518 and TOPPGene online tools , we found fifty-four genes common to the three prioritization tools which formed our candidate leukemogenic genes for this study . As per the protocol we selected thirty training genes from PubMed . The prioritized and training genes were then used to construct STRING functional association network , which was further analyzed using cytoHubba hub analysis tool to investigate new genes which could form drug targets in leukemia . Analysis of the STRING protein network built from these prioritized and training genes led to identification of two hub genes , Q15796 and P50750 , which were not implicated in leukemogenesis earlier . Filtering out from several hundred genes in the network we also found O00255 , Q13547 and P06239 genes , which re-emphasized the important role of these genes in leukemogenesis . This is the first report on these five additional signature genes in leukemogenesis . We propose these as new targets for developing novel therapeutics and also as biomarkers in leukemogenesis , which could be important for prognosis and diagnosis . Genetic underpinnings of tardive dyskinesia : passing the baton to pharmacogenetics . Manifestation of tardive dyskinesia ( TD ) among schizophrenia subjects on long-term antipsychotic treatment with typical drugs has been a clinical concern . Despite its association with extrapyramidal symptoms , typical drugs are still routinely prescribed globally though marginally superior atypical drugs have long been available . The genetic component in the etiology of TD is well documented . Search for these determinants has led to a few consensus associations of P10635 *10 , P05177 *1F , P14416 Taq1A ( rs1800497 ) , P35462 Ser9Gly ( rs6280 ) and MnSOD Ala9Val ( rs4880 ) variants with TD . However , translation of these observations into the clinic has not been achieved so far . This review discusses the salient features of TD etiopathology , current status of TD genetics , interactions between genetic and nongenetic factors , some major drawbacks , challenges and expected focus in TD research over the next decade , with emphasis on pharmacogenetics . 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 . 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 . 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 . 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 . Serotonin 5-hT1A receptor activation prevents phosphorylation of DB01221 receptor Q9UHB4 subunit in cerebral ischemia . The mechanisms involved in the neuroprotective effect of serotonin P08908 receptor agonists on brain damage induced by ischemia remain to be fully elucidated . Given that serotonergic drugs may regulate N-methyl-D-aspartate ( DB01221 ) receptor function , which is implicated in events leading to ischemia-induced neuronal cell death , this study sought to determine the effects of the selective P08908 receptor agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , on the levels of DB01221 receptor Q9UHB4 subunit in gerbil hippocampus after transient global cerebral ischemia . Pretreatment with 8-OH-DPAT ( 1 mg/kg ) prevented the neuronal loss in P00915 subfield 72 h after ischemia . DB01221 receptor Q9UHB4 levels in whole hippocampus were not affected 24 h after ischemia , but the levels of the subunit phosphorylated at the protein kinase A ( PKA ) site , pNR1(Ser897) , were significantly increased , and this increase was prevented by the same 8-OH-DPAT dose , a probable consequence of the increased phosphatase 1 ( P50391 ) enzyme activity found in ischemic gerbils pretreated with the P08908 receptor agonist . The results suggest that Q9UHB4 subunit phosphorylation plays a role in the neuroprotective effect of 8-OH-DPAT on cell damage induced by global cerebral ischemia in the gerbil hippocampus and support the potential interest of P08908 receptor activation in the search for neuroprotective strategies . 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 . 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 . 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 . 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 . DB00989 improves hippocampal neurogenesis and depression-like behaviors via P08908 receptor stimulation in olfactory bulbectomized mice . DB00989 is a non-competitive inhibitor of both acetylcholinesterase ( P22303 ) and butylcholinesterase ( BuChE ) used to treat mild to moderate dementia in Alzheimer 's disease ( AD ) patients . Although rivastigmine reportedly ameliorates cognitive dysfunction in these patients , its ability to improve Behavioral and Psychological Symptoms of Dementia ( BPSD ) remains unclear . To determine whether rivastigmine treatment antagonizes depression-like behaviors , we chronically administered rivastigmine ( 0.1-1.0mg/kg ) to olfactory bulbectomized ( OBX ) mice once a day for 2weeks , starting 2weeks after bulbectomy . Chronic treatment at 0.3 or 1.0mg/kg dose dependently and significantly improved depression-like behaviors , as assessed by tail suspension ( Q16762 ) , forced swim ( P19883 ) , locomotion and novelty-suppressed feeding ( NSFT ) tests . Importantly , co-administration with WAY-100635 ( 1.0mg/kg ) , a P08908 receptor antagonist , but not ketanserin ( 1.0mg/kg , ) , a 5- Q13049 receptor antagonist , completely blocked rivastigmine-induced anti-depressive effects , suggesting that P08908 receptor stimulation mediates this activity . Consistent with this observation , rivastigmine treatment significantly rescued impaired neurogenesis observed in OBX mice in a P08908 receptor-dependent manner . Furthermore , enhanced protein kinase B ( Akt ) and extracellular signal-regulated kinase ( P29323 ) phosphorylation seen following rivastigmine treatment was closely associated with improved neurogenesis . These effects were blocked by WAY-100635 but not ketanserin treatment . Finally , we confirmed that P08908 but not 5- Q13049 receptor stimulation by specific agonists mimicked rivastigmine-induced anti-depression activity and promoted hippocampal neurogenesis . We conclude that , in addition to enhancing the cholinergic system , rivastigmine treatment restores normal function of the hippocampal serotonergic system , an activity that likely ameliorates depressive behaviors in AD patients . 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 . 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 . 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 . Ellagic acid inhibits P15692 / P35968 , PI3K/Akt and MAPK signaling cascades in the hamster cheek pouch carcinogenesis model . BACKGROUND : Blocking vascular endothelial growth factor ( P15692 ) mediated tumor angiogenesis by phytochemicals has emerged as an attractive strategy for cancer prevention and therapy . METHODS : We investigated the anti-angiogenic effects of ellagic acid in a hamster model of oral oncogenesis by examining the transcript and protein expression of hypoxia-inducible factor-1alpha ( HIF-1α ) , P15692 , P35968 , and the members of the PI3K/Akt and MAPK signaling cascades . Molecular docking studies and cell culture experiments with the endothelial cell line ECV304 were performed to delineate the mechanism by which ellagic acid regulates P15692 signaling . RESULTS : We found that ellagic acid significantly inhibits HIF-1α-induced P15692 / P35968 signalling in the hamster buccal pouch by abrogating PI3K/Akt and MAPK signaling via downregulation of PI3K , PDK-1 , p-Akt(ser473) , P42345 , p- P29323 , and p-JNK . Ellagic acid was also found to reduce the expression of histone deacetylases that could inhibit neovascularization . Analysis of the mechanism revealed that ellagic acid inhibits hypoxia-induced angiogenesis via suppression of HDAC-6 in ECV304 cells . Furthermore , knockdown of endogenous Q9UBN7 via small interfering RNA abrogated hypoxia-induced expression of HIF-1α and P15692 and blocked Akt activation . Molecular docking studies confirmed interaction of ellagic acid with upstream kinases that regulate angiogenic signaling . CONCLUSIONS : Taken together , these findings demonstrate that the anti-angiogenic activity of ellagic acid may be mediated by abrogation of hypoxia driven PI3K/Akt/ P42345 , MAPK and P15692 / P35968 signaling pathways involving suppression of Q9UBN7 and HIF-1α responses . GENERAL SIGNIFICANCE : Ellagic acid offers promise as a lead compound for anticancer therapeutics by virtue of its ability to inhibit key oncogenic signaling cascades and HDACs . DBC1/ Q8N163 and Q8IX12 Are Largely Disordered Proteins that Have Evolved from One Common Ancestor . Deleted in breast cancer 1 ( DBC1 , Q8N163 , Q8N163 ) is a large , predominantly nuclear , multidomain protein that modulates gene expression by inhibiting several epigenetic modifiers , including the deacetylases Q96EB6 and O15379 , and the methyltransferase O43463 . DBC1 shares many highly conserved protein domains with its paralog cell cycle and apoptosis regulator 1 ( Q8IX12 , CARP-1 ) . In this study , we examined the full-length sequential and structural properties of DBC1 and Q8IX12 from multiple species and correlated these properties with evolution . Our data shows that the conserved domains shared between DBC1 and Q8IX12 have similar domain structures , as well as similar patterns of predicted disorder in less-conserved intrinsically disordered regions . Our analysis indicates similarities between DBC1 , Q8IX12 , and the nematode protein lateral signaling target 3 ( G3V0H7 ) , suggesting that DBC1 and Q8IX12 may have evolved from G3V0H7 . Our data also suggests that DBC1 emerged later in evolution than Q8IX12 . DBC1 contains regions that show less conservation across species as compared to the same regions in Q8IX12 , suggesting a continuously evolving scenario for DBC1 . Overall , this study provides insight into the structure and evolution of DBC1 and Q8IX12 , which may impact future studies on the biological functions of these proteins . 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 . 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 . [ 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 . P35712 suppresses cyclin D1 promoter activity by interacting with beta-catenin and histone deacetylase 1 , and its down-regulation induces pancreatic beta-cell proliferation . Sex-determining region Y-box ( SOX ) 6 negatively regulates glucose-stimulated insulin secretion from beta-cells and is a down-regulated transcription factor in the pancreatic islet cells of hyperinsulinemic obese mice . To determine the contribution of P35712 to insulin resistance , we analyzed the effects of P35712 on cell proliferation . Small interfering RNA-mediated attenuation of P35712 expression stimulated the proliferation of insulinoma P01308 -1E and NIH-3T3 cells , whereas retroviral overexpression resulted in inhibition of cell growth . Quantitative real time-PCR analysis revealed that the levels of cyclin D1 transcripts were markedly decreased by P35712 overexpression . Luciferase-reporter assay with beta-catenin showed that P35712 suppresses cyclin D1 promoter activities . In vitro binding experiments showed that the LZ/Q domain of P35712 physically interacts with armadillo repeats 1-4 of beta-catenin . Furthermore , chromatin immunoprecipitation assay revealed that increased P35712 expression significantly reduced the levels of acetylated histones H3 and H4 at the cyclin D1 promoter . By using a histone deacetylase ( HDAC ) inhibitor and co-immunoprecipitation analysis , we showed that P35712 suppressed cyclin D1 activities by interacting withbeta-catenin and Q13547 . The data presented suggest that P35712 may be an important factor in obesity-related insulin resistance . The cleaved cytoplasmic tail of polycystin-1 regulates Src-dependent P40763 activation . P98161 ( PC1 ) mutations result in proliferative renal cyst growth and progression to renal failure in autosomal dominant polycystic kidney disease ( ADPKD ) . The transcription factor P40763 ( signal transducer and activator of transcription 3 ) was shown to be activated in cyst-lining cells in ADPKD and Q15139 mouse models and may drive renal cyst growth , but the mechanisms leading to persistent P40763 activation are unknown . A proteolytic fragment of PC1 corresponding to the cytoplasmic tail , PC1-p30 , is overexpressed in ADPKD . Here , we show that PC1-p30 interacts with the nonreceptor tyrosine kinase Src , resulting in Src-dependent activation of P40763 by tyrosine phosphorylation . The PC1-p30-mediated activation of Src/ P40763 was independent of JAK family kinases and insensitive to the P40763 inhibitor suppressor of cytokine signaling 3 . Signaling by the P01133 receptor ( P00533 ) or DB02527 amplified the activation of Src/ P40763 by PC1-p30 . Expression of PC1-p30 changed the cellular response to DB02527 signaling . In the absence of PC1-p30 , DB02527 dampened P00533 - or P05231 -dependent activation of P40763 ; in the presence of PC1-p30 , DB02527 amplified Src-dependent activation of P40763 . In the polycystic kidney ( PCK ) rat model , activation of P40763 in renal cystic cells depended on vasopressin receptor 2 ( P30518 ) signaling , which increased DB02527 levels . Genetic inhibition of vasopressin expression or treatment with a pharmacologic P30518 inhibitor strongly suppressed P40763 activation and reduced renal cyst growth . These results suggest that PC1 , via its cleaved cytoplasmic tail , integrates signaling inputs from P00533 and DB02527 , resulting in Src-dependent activation of P40763 and a proliferative response . Molecular mechanisms of the antiangiogenic and antitumor effects of mycophenolic acid . The relative risk for the development of malignancies following solid organ transplantation seems to be decreased in patients treated with the immunosuppressive agent mycophenolic acid ( DB00603 ) . However , the molecular mechanisms of the antineoplastic effects of DB00603 are not completely understood . Here , we report that human endothelial cells and fibroblasts are highly sensitive to DB00603 treatment . We found that U87 glioblastoma cells were resistant to DB00603 treatment in vitro . However , U87 tumor growth was markedly inhibited in vivo in BALB/c nude mice , suggesting that DB00603 exerted its antitumor effects via modulation of the tumor microenvironment . Accordingly , microvascular density and pericyte coverage were markedly reduced in DB00603 -treated tumors in vivo . Using functional in vitro assays , we showed that DB00603 potently inhibited endothelial cell and fibroblast proliferation , invasion/migration , and endothelial cell tube formation . To identify the genetic participants governing the antiangiogenic and antifibrotic effects of DB00603 , we performed genome-wide transcriptional analysis in U87 , endothelial and fibroblast cells at 6 and 12 h after DB00603 treatment . Network analysis revealed a critical role for MYC signaling in endothelial cells treated with DB00603 . Moreover , we found that the antiangiogenic effects of DB00603 were organized by coordinated communications between MYC and Q92597 , YYI , Q16665 , Q92769 , P06493 , P49841 , and P22694 signaling . The regulation of these " hub nodes " was confirmed by real-time quantitative reverse transcription-PCR and protein analysis . The critical involvement of MYC in the antiangiogenic signaling of DB00603 was further shown by gene knockdown experiments . Together , these data provide a molecular basis for the antiangiogenic and antifibrotic effects of DB00603 , which warrants further clinical investigations . 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 . 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 . Dopamine receptors and psychiatric drug treatment . The established antipsychotic drugs act mainly by antagonizing dopamine mediated synaptic transmission in the brain . Increase in the rate of production of dopamine metabolites as well as the firing rate of dopamine-containing neurons can be interpreted as compensatory responses to an interruption of synaptic transmission at dopamine nerve terminals . The demonstration of involvement of limbic and cortical mechanisms in the antipsychotic activity of neuroleptic drugs is far more difficult than the involvement of nigro-striatal and tubero-infundibular mechanisms in the neurological and neuroendocrine effects of these drugs . Application of radioreceptor techniques to dopamine research has supported the findings obtained by other neuropsychopharmacological research techniques , providing more direct evidence of dopamine receptor blockade by neuroleptic drugs . Further research is needed especially in studying the nature of the time-dependent adaptive changes at the receptor sites as well as the differences between the different dopamine projections and neural systems in the brain . The different subtypes of dopamine receptors in the brain , currently called D1 and D2 dopamine receptors , seem to be parallel , although in many respects independently-acting regulatory systems . P14416 -selective antagonists such as sulpiride seem to cause selective D2 receptor up-regulation . P01236 secretion seems to be regulated by D2 dopamine receptors . The exact physiological role of D1 dopamine receptors as well as the clinical consequences of selective D1 antagonism is not known . DB00391 and clozapine are examples of atypical neuroleptic compounds that have quite different profile of action , the former having strong and selective antidopaminergic action , the latter combining a number of non- dopaminergic mechanisms with rather slight effects on dopamine receptors. ( ABSTRACT TRUNCATED AT 250 WORDS ) Q16696 enhances the sensitivity of human bronchial epithelial cells to aflatoxin B1-induced DNA damage . Q16696 ( Q16696 ) mainly expresses in human respiratory system and mediates the metabolic activation of aflatoxin B1 ( AFB1 ) . Our previous study suggested that Q16696 could increase the cytotoxic and apoptotic effects of AFB1 in immortalized human bronchial epithelial cells ( BEAS-2B ) . However , the role of Q16696 in AFB1-induced DNA damage is unclear . Using BEAS-2B cells that stably express Q16696 ( B-2A13 ) , P05177 ( B-1A2 ) , and P11509 ( B-2A6 ) , we compared their effects in AFB1-induced DNA adducts , DNA damage , and cell cycle changes . BEAS-2B cells that were transfected with vector ( B-vector ) were used as a control . The results showed that AFB1 ( 5-80 nM ) dose- and time-dependently induced DNA damage in B-2A13 cells . AFB1 at 10 and 80nM significantly augmented this effect in B-2A13 and B-1A2 cells , respectively . B-2A6 cells showed no obvious DNA damage , similar to B-vector cells and the vehicle control . Similarly , compared with B-vector , B-1A2 or B-2A6 cells , B-2A13 cells showed more sensitivity in AFB1-induced γ P16104 expression , DNA adduct 8-hydroxy-deoxyguanosine formation , and S-phase cell-cycle arrest . Furthermore , AFB1 activated the proteins related to DNA damage responses , such as Q13315 , ATR , Chk2 , p53 , P38398 , and P16104 , rather than the proteins related to DNA repair . These effects could be almost completely inhibited by 100 μM nicotine ( a substrate of Q16696 ) or 1 μM 8-methoxypsoralen ( DB00553 ; an inhibitor of CYP enzyme ) . Collectively , these findings suggest that Q16696 plays an important role in low-concentration AFB1-induced DNA damage , possibly linking environmental airborne AFB1 to genetic injury in human respiratory system . 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 .

MH_train_1135

MH_train_1135

MH_train_1135

interacts_with DB00898?

multiple_choice

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 . 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 . 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 . Evidence of an Epigenetic Modification in Cell-cycle Arrest Caused by the Use of Ultra-highly-diluted Gonolobus Condurango Extract . OBJECTIVES : Whether the ultra-highly-diluted remedies used in homeopathy can effectively bring about modulations of gene expressions through acetylation/deacetylation of histones has not been explored . Therefore , in this study , we pointedly checked if the homeopathically-diluted anti-cancer remedy Condurango 30C ( ethanolic extract of Gonolobus condurango diluted 10(-60) times ) was capable of arresting the cell cycles in cervical cancer cells HeLa by triggering an epigenetic modification through modulation of the activity of the key enzyme histone deacetylase 2 vis-a-vis the succussed alcohol ( placebo ) control . METHODS : We checked the activity of different signal proteins ( like P38936 (WAF) , p53 , Akt , P40763 ) related to deacetylation , cell growth and differentiation by western blotting and analyzed cell-cycle arrest , if any , by fluorescence activated cell sorting . After viability assays had been performed with Condurango 30C and with a placebo , the activities of histone de-acetylase ( HDAC ) enzymes 1 and 2 were measured colorimetrically . RESULTS : While Condurango 30C induced cytotoxicity in HeLa cells in vitro and reduced Q92769 activity quite strikingly , it apparently did not alter the Q13547 enzyme ; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity . Data on P38936 (WAF) , p53 , Akt , and P40763 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and P55008 -phase cell-cycle arrest when Condurango 30C was used at a 2 % dose . CONCLUSION : Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells , which the placebo was unable to do . μ-Opioid receptor attenuates Aβ oligomers-induced neurotoxicity through P42345 signaling . AIMS : μ-opioid receptor ( P35372 ) exerts many functions such as antinociception , neuroprotection , and hippocampal plasticity . A body of evidence has shown that P35372 activation could stimulate downstream effectors of mechanistic/mammalian target of rapamycin ( P42345 ) . However , it is not clear whether P35372 protects neurons against β-amyloid peptide ( Aβ ) neurotoxicity through P42345 signaling . METHODS : The effects of P35372 activation on Aβ oligomers-induced neurotoxicity were assessed by cell viability and neurite outgrowth assay in primary cultured cortical neurons . The activities of P42345 , protein kinase B ( Akt ) and P08133 ribosomal S6 kinase ( P08133 S6k ) upon P35372 activation by morphine were measured by immunoblotting their phosphorylation status . RESULTS : Morphine dose-dependently attenuated Aβ oligomers-induced neurotoxicity . Aβ oligomers downregulated P42345 signaling . Morphine significantly rescued P42345 signaling by reversal of Aβ oligomers ' effect on P42345 and its upstream signaling molecule Akt , as well as its downstream molecule P08133 S6k . Moreover , the neuroprotective effect of morphine could be reversed by P35372 selective antagonist and phosphatidylinositol 3-kinases ( PI3K ) , Akt and P42345 inhibitors . Furthermore , endogenous opioids-enkaphalins also attenuated Aβ oligomers-induced neurotoxicity . CONCLUSIONS : Our findings demonstrated P35372 activation attenuated Aβ oligomers-induced neurotoxicity through P42345 signaling . It may provide new insight into the pathological process and useful strategy for therapeutic interventions against Aβ neurotoxicity . Immunocytochemical localization of Q9UBS5 receptor subunits in the basolateral amygdala . Gamma-aminobutyric acid B ( GABAB ) receptors ( GBRs ) are G-protein-coupled receptors that mediate a slow , prolonged form of inhibition in the basolateral amygdala ( P00519 ) and other brain areas . Recent studies indicate that this receptor is a heterodimer consisting of Q9UBS5 ( GBR1 ) and O75899 subunits . In the present investigation , antibodies to the Q9UBS5 subunit were used to study the neuronal localization of GBRs in the rat P00519 . GBR immunoreactivity was mainly found in spine-sparse interneurons and astrocytes at the light microscopic level . Very few pyramidal neurons exhibited perikaryal staining . Dual-labeling immunofluorescence analysis indicated that each of the four main subpopulations of interneurons exhibited GBR immunoreactivity . Virtually 100 % of large CCK+ neurons in the basolateral and lateral nuclei were GBR+ . In the basolateral nucleus 72 % of somatostatin ( Q8TE85 ) , 73 % of parvalbumin ( PV ) and 25 % of P01282 positive interneurons were GBR+ . In the lateral nucleus 50 % of somatostatin , 30 % of parvalbumin and 27 % of P01282 positive interneurons were GBR+ . Electron microscopic ( EM ) analysis revealed that most of the light neuropil staining seen at the light microscopic level was due to the staining of dendritic shafts and spines , most of which probably belonged to spiny pyramidal cells . Very few axon terminals ( Ats ) were GBR+ . In summary , this investigation demonstrates that the distal dendrites of pyramidal cells , and varying percentages of each of the four main subpopulations of interneurons in the P00519 , express GBRs . Because previous studies suggest that GBR-mediated inhibition modulates DB01221 -dependent EPSPs in the P00519 , these receptors may play an important role in neuronal plasticity related to emotional learning . 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 . 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 . Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking . Binge drinking of alcoholic beverages among adolescents is a common practice that can have serious health consequences . DB00898 is a potent immunomodulator that alters a wide range of immune responses . However , it is unclear whether there is a differential immune response to alcoholic beverages with a high versus low concentration of ethanol . In this study , we used a PCR array containing 46 primer pairs of selected genes to compare mRNA expression in the spleen , an immune system organ , of adolescent rats following binge drinking of alcohol solutions containing either 20 % or 52 % ethanol ( v/v , 4.8 g/kg daily dosage ) , or water ( control ) for 3 d . We found that , expression of IL-1β , P05231 , P13500 , and GABA(A) receptor α2 subunit in the spleen were decreased , and P41594 and P46098 receptor expression were increased after administration of an ethanol solution containing 52 % ethanol , but not one with 20 % ethanol . Our data suggest that alcohol-mediated immunomodulatory effects are , in part , dependent on the ethanol by volume concentration . This is the first study to show that exposure to a high ethanol percentage beverage can have more profound effects on immune responses than one with a low percentage of ethanol . P04626 interacts with P16070 to up-regulate P61073 via epigenetic silencing of microRNA-139 in gastric cancer cells . BACKGROUND & AIMS : Human epidermal growth factor receptor 2 ( P04626 ) ( neu/ P04626 ) is overexpressed on many types of cancer cells , including gastric cancer cells ; P04626 overexpression has been associated with metastasis and poor prognosis . We investigated the mechanisms by which P04626 regulates cell migration and invasion . METHODS : P04626 expression or activity was reduced in gastric cancer cell lines using small interfering RNAs or the monoclonal antibody , trastuzumab . We identified proteins that interact with P04626 or microRNAs ( miRNAs ) involved in P04626 signaling . We used various software programs to identify miRNAs that regulate factors in the P04626 signaling pathway . We analyzed expression patterns of these miRNAs in gastric cancer cell lines and tumor samples from patients . RESULTS : We found that P16070 binds directly to P04626 , which up-regulates the expression of metastasis-associated protein-1 , induces deacetylation of histone H3 lysine 9 , and suppresses transcription of microRNA139 ( miR-139 ) to inhibit expression of its target gene , P61073 ( P61073 ) . Knockdown of P04626 and P16070 reduced invasive activity of cultured gastric cancer cells and suppressed tumor growth in nude mice . Lymph node metastasis was associated with high levels of P04626 , P16070 , and P61073 , and reduced levels of miR-139 in human metastatic gastric tumors . Cultures of different types of metastatic cancer cells with histone deacetylase inhibitors and/or DNA methyltransferase resulted in up-regulation of miR-139 . CONCLUSIONS : P04626 interaction with P16070 up-regulates P61073 by inhibiting expression of miR-139 , at the epigenetic level , in gastric cancer cells . These findings indicate how P04626 signaling might promote gastric tumor progression and metastasis . Selective targeting of the repressive transcription factors P25490 and cMyc to disrupt quiescent human immunodeficiency viruses . Quiescent HIV-1 infection of resting P01730 (+) T cells is an obstacle to eradication of HIV-1 infection . These reservoirs are maintained , in part , by repressive complexes that bind to the HIV-1 long terminal repeat ( LTR ) and recruit histone deacetylases (HDACs). cMyc and P25490 are two transcription factors that are recruited as part of well-described , distinct complexes to the HIV-1 LTR and in turn recruit HDACs . In prior studies , depletion of single factors that recruit Q13547 in various cell lines was sufficient to upregulate LTR activity . We used short hairpin RNAs ( shRNAs ) to test the effect of targeted disruption of a single transcription factor on quiescent proviruses in T cell lines . In this study , we found that depletion of P25490 significantly increases mRNA and protein expression from the HIV-1 promoter in some contexts , but does not affect Q13547 , Q92769 , O15379 , or acetylated histone 3 occupancy of the HIV-1 LTR . Conversely , depletion of cMyc or cMyc and P25490 does not significantly alter the level of transcription from the LTR or affect recruitment of HDACs to the HIV-1 LTR . Furthermore , global inhibition of HDACs with the HDAC inhibitor suberoylanilide hydroxamic acid ( DB02546 ) enhanced the increase in LTR transcription in cells that were depleted of P25490 .These findings show that despite prior isolated findings , redundancy in repressors of HIV-1 LTR expression will require selective targeting of multiple restrictive mechanisms to comprehensively induce the escape of quiescent proviruses from latency . 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 . [ 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 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 . GABA Receptors Genes Polymorphisms and DB00898 Dependence : No Evidence of an Association in an Italian Male Population . OBJECTIVE : The genes encoding for gamma-aminobutyric acid ( GABA ) A and B receptors may be considered as candidates for alcoholism ; genetic alterations at this level may produce structural and functional diversity and thus play a role in the response to alcohol addiction treatment . To investigate these aspects further , we conducted a preliminary genetic association study on a population of Italian male alcohol addicts , focusing on GABA A and B receptors . METHODS : A total of 186 alcohol-dependent subjects ( in the first phase 139 , then 47 more samples ) and 182 controls were genotyped for 25 single nucleotide polymorphisms ( SNPs ) of genes encoding the alpha-1 subunit of GABA A receptor ( P14867 ) and subunits 1 and 2 of GABA B receptor ( Q9UBS5 and O75899 ) . The chi-squared test for allele and genotype distributions and Hardy-Weinberg equilibrium analysis of both subjects and controls were performed . Bonferroni 's correction for multiple comparisons was applied . RESULTS : Preliminary results comparing 139 alcohol-dependent subjects and 182 controls showed differences in genotype distribution in the former for SNP rs29253 , located in the intron region of the Q9UBS5 gene . In order to clarify the meaning of this association , 47 more samples from alcohol-dependent subjects were tested for this SNP only : the previously found association was not confirmed . CONCLUSION : The lack of significant differences between the two groups does not provide evidence that GABRA 1 and Q9UBS5 and 2 genes are candidates for alcoholism in this population . Further studies with larger samples are needed , together with investigation of other components of the GABA pathway . 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 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 . 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 . Nicotinic acetylcholine receptors containing the α6 subunit contribute to ethanol activation of ventral tegmental area dopaminergic neurons . DB00184 and alcohol are often co-abused suggesting a common mechanism of action may underlie their reinforcing properties . Both drugs acutely increase activity of ventral tegmental area ( VTA ) dopaminergic ( DAergic ) neurons , a phenomenon associated with reward behavior . Recent evidence indicates that nicotinic acetylcholine receptors ( nAChRs ) , ligand-gated cation channels activated by ACh and nicotine , may contribute to ethanol-mediated activation of VTA DAergic neurons although the nAChR subtype(s) involved has not been fully elucidated . Here we show that expression and activation of nAChRs containing the α6 subunit contribute to ethanol-induced activation of VTA DAergic neurons . In wild-type ( WT ) mouse midbrain sections that contain the VTA , ethanol ( 50 or 100 mM ) significantly increased firing frequency of DAergic neurons . In contrast , ethanol did not significantly increase activity of VTA DAergic neurons in mice that do not express Q15825 , the gene encoding the α6 nAChR subunit ( α6 knock-out ( KO ) mice ) . DB00898 -induced activity in WT slices was also reduced by pre-application of the α6 subtype-selective nAChR antagonist , α-conotoxin MII[E11A] . When co-applied , ethanol potentiated the response to ACh in WT DAergic neurons ; whereas co-application of ACh and ethanol failed to significantly increase activity of DAergic neurons in α6 KO slices . Finally , pre-application of α-conotoxin MII[E11A] in WT slices reduced ethanol potentiation of ACh responses . Together our data indicate that α6-subunit containing nAChRs may contribute to ethanol activation of VTA DAergic neurons . These receptors are predominantly expressed in DAergic neurons and known to be critical for nicotine reinforcement , providing a potential common therapeutic molecular target to reduce nicotine and alcohol co-abuse . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Design , synthesis and biological evaluation of pazopanib derivatives as antitumor agents . A novel series of pazopanib derivatives were designed , synthesized , and evaluated for their inhibitory activity against a series of kinases including P35968 , P00533 , P31749 , P37023 , and P00519 . The anti-angiogenic activities ex vivo of some compounds were also investigated . Compounds P2d and P2e demonstrated outstanding inhibitory activity against P35968 and P00519 and higher anti-angiogenic activity compared with DB06589 , the reference standard . These two compounds ( P2d and P2e ) could be used as novel lead compounds for further development of anticancer agents . 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 ) . 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 . 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 . 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 . The kinase inhibitor TKI258 is active against the novel CUX1- P11362 fusion detected in a patient with T-lymphoblastic leukemia/lymphoma and t(7;8)(q22;p11) . We report a patient with T-lymphoblastic leukemia/lymphoma and a t(7;8)(q22;p11) . CUX1 was identified as the fusion partner of P11362 by fluorescence in situ hybridization and 5' RACE-PCR . We further investigated this novel P11362 fusion using the interleukin-3 ( P08700 ) dependent Ba/ P13726 cell line and demonstrated P08700 independent cell growth of CUX1- P11362 expressing cells . TKI258 and PKC412 potently inhibited proliferation of CUX1- P11362 transformed Ba/ P13726 cells . This growth inhibition was shown to be mediated by inhibition of CUX1- P11362 kinase activity for TKI258 but not PKC412 . In summary , we identified a novel CUX1- P11362 fusion oncogene in a patient with the 8p11 myeloproliferative syndrome and demonstrated its transforming potential in the Ba/ P13726 cell line . Our in vitro data support the further investigation of TKI258 for the treatment of constitutively active P11362 fusion proteins . 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 . 17β-estradiol inhibits P14780 and Q09428 /TrpM4 expression and activation and thereby attenuates BSCB disruption/hemorrhage after spinal cord injury in male rats . Blood-spinal cord barrier ( BSCB ) disruption and progressive hemorrhage after spinal cord injury ( SCI ) lead to secondary injury and the subsequent apoptosis and/or necrosis of neuron and glia , causing permanent neurological deficits . In this study , we examined the effect of 17β-estradiol ( E2 ) on BSCB breakdown and hemorrhage as well as subsequent inflammation after SCI . After a moderate contusion injury at the 9th thoracic segment of spinal cord , E2 ( 300 μg/kg ) was administered by iv injection immediately after SCI , and the same dose of E2 was then administered 6 and 24 hours after injury . Our data show that E2 attenuated BSCB permeability and hemorrhage and reduced the infiltration of neutrophils and macorphages after SCI . Consistent with this finding , the expression of inflammatory mediators was significantly reduced by E2 . Furthermore , E2 treatment significantly inhibited the expression of sulfonylurea receptor 1 and transient receptor potential melastatin 4 after injury , which are known to mediate hemorrhage at an early stage after SCI . Moreover , the expression and activation of matrix metalloprotease-9 after injury , which is known to disrupt BSCB , and the degradation of tight junction proteins , such as zona occludens-1 and occludin , were significantly inhibited by E2 treatment . Furthermore , the protective effects of E2 on BSCB disruption and functional improvement were abolished by an estrogen receptor antagonist , ICI 182780 ( 3 mg/kg ) . Thus , our study provides evidence that the neuroprotective effect of E2 after SCI is , in part , mediated by inhibiting BSCB disruption and hemorrhage through the down-regulation of sulfonylurea receptor 1/transient receptor potential melastatin 4 and matrix metalloprotease-9 , which is dependent on estrogen receptor . 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 . Polymorphisms influencing olanzapine metabolism and adverse effects in healthy subjects . OBJECTIVE : The pharmacokinetics of olanzapine and response to treatment could be affected by polymorphisms in genes coding for drug-metabolizing enzymes , transporters , or receptors . The aim of this study was to identify genetic markers predictive of pharmacokinetics , pharmacodynamics , and adverse effects of olanzapine . METHODS : Sixty-three healthy volunteers receiving a single 5-mg oral dose of olanzapine were genotyped for 39 genetic variants that could be related to the response to olanzapine . All genetic variants were analyzed by PharmaChip , but P14416 Taq1A polymorphism was determined by real-time polymerase chain reaction . Olanzapine was measured using high-performance liquid chromatography combined with tandem mass spectrometry . The relationship of gender and polymorphisms with olanzapine pharmacokinetics , the change in prolactin levels , and the incidence of adverse effects were evaluated by multiple regression analysis . RESULTS : The pharmacokinetics of olanzapine was influenced by polymorphisms in P20815 , P21266 , and Q13224 . P01236 levels were affected by gender and polymorphisms in P14416 and 5- P28223 . Polymorphisms in P11712 , P51580 , P22309 , P08183 , and 5- P28223 were related to some adverse effects of olanzapine . CONCLUSIONS : Several polymorphisms can explain differences in the pharmacokinetics , pharmacodynamics , and safety of olanzapine in healthy subjects . Whether these genetic factors influence the risk of therapeutic failure or tolerability in patients remains to be established . GSK3 controls axon growth via CLASP-mediated regulation of growth cone microtubules . Suppression of glycogen synthase kinase 3 ( GSK3 ) activity in neurons yields pleiotropic outcomes , causing both axon growth promotion and inhibition . Previous studies have suggested that specific GSK3 substrates , such as adenomatous polyposis coli ( P25054 ) and collapsin response mediator protein 2 ( Q16555 ) , support axon growth by regulating the stability of axonal microtubules ( MTs ) , but the substrate(s) and mechanisms conveying axon growth inhibition remain elusive . Here we show that CLIP ( cytoplasmic linker protein ) -associated protein ( CLASP ) , originally identified as a MT plus end-binding protein , displays both plus end-binding and lattice-binding activities in nerve growth cones , and reveal that the two MT-binding activities regulate axon growth in an opposing manner : The lattice-binding activity mediates axon growth inhibition induced by suppression of GSK3 activity via preventing MT protrusion into the growth cone periphery , whereas the plus end-binding property supports axon extension via stabilizing the growing ends of axonal MTs . We propose a model in which CLASP transduces GSK3 activity levels to differentially control axon growth by coordinating the stability and configuration of growth cone MTs . 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 . Complex formation with the Type B gamma-aminobutyric acid receptor affects the expression and signal transduction of the extracellular calcium-sensing receptor . Studies with P29320 -293 cells and neurons . We co-immunoprecipitated the Ca(2+)-sensing receptor ( CaR ) and type B gamma-aminobutyric acid receptor ( GABA-B-R ) from human embryonic kidney ( P29320 ) -293 cells expressing these receptors and from brain lysates where both receptors are present . CaRs extensively co-localized with the two subunits of the GABA-B-R ( Q96GN5 and R2 ) in P29320 -293 cell membranes and intracellular organelles . Coexpressing CaRs and GABA-B-R1s in P29320 -293 cells suppressed the total cellular and cell surface expression of CaRs and inhibited phospholipase C activation in response to high extracellular [ Ca(2+) ] ( [Ca(2+)](e) ) . In contrast , coexpressing CaRs and GABA-B-R2s enhanced CaR expression and signaling responses to raising [Ca(2+)](e) . The latter effects of the O75899 on the CaR were blunted by coexpressing the Q9UBS5 . Coexpressing the CaR with Q9UBS5 or R2 enhanced the total cellular and cell surface expression of the Q9UBS5 or R2 , respectively . Studies with truncated CaRs indicated that the N-terminal extracellular domain of the CaR participated in the interaction of the CaR with the Q9UBS5 and R2 . In cultured mouse hippocampal neurons , CaRs co-localized with the Q9UBS5 and R2 . CaRs and GABA-B-R1s also co-immunoprecipitated from brain lysates . The expression of the CaR was increased in lysates from Q9UBS5 knock-out mouse brains and in cultured hippocampal neurons with their Q9UBS5 genes deleted in vitro . Thus , CaRs and GABA-B-R subunits can form heteromeric complexes in cells , and their interactions affect cell surface expression and signaling of CaR , which may contribute to extracellular Ca(2+)-dependent receptor activation in target tissues . 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 . Low ethanol concentration alters P30532 RNA levels during early human development . DB00898 use is common and consumption during pregnancy has been shown to lead to a myriad of physical and neurologic abnormalities commonly referred to as fetal alcohol spectrum disorder . Substance addiction , which includes alcohol , has been shown to involve the major nicotinic acetylcholine receptor subunit P30532 . Using human embryonic stem cells as a model of early human development , we show that low concentrations of ethanol ( 20mM ) can alter the expression of P30532 . Changes in P30532 expression is linked to altered GABA and DB01221 receptor expression , as well as abnormal development of the frontal cortex . These results suggest that alcohol exposure can alter early neurologic development , which may favor addiction and other developmental abnormalities in unborn children . 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 . 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 . Dual P00533 and P42345 targeting in squamous cell carcinoma models , and development of early markers of efficacy . The epidermal growth factor receptor ( P00533 ) is a validated target in squamous cell carcinoma ( SCC ) of the head and neck . Most patients , however , do not respond or develop resistance to this agent . P42345 ( P42345 ) is involved in the pathogenesis of SCC of the head and neck ( SCCHN ) . This study aimed to determine if targeting P42345 in combination with P00533 is effective in SCC , and to develop early pharmacodynamic markers of efficacy . Two SCC cell lines , one resistant ( HEP2 ) and one of intermediate susceptibility ( Detroit 562 ) to P00533 inhibitors , were xenografted in vivo and treated with an P42345 inhibitor ( temsirolimus ) , an P00533 inhibitor ( erlotinib ) or a combination of both . DB06287 exerted superior growth arrest in both cell lines than erlotinib . The combined treatment resulted in synergistic antitumor effects in the Detroit 562 cell line . Immunohistochemical assessment of pharmacodynamic effects in fine-needle aspiration ( FNA ) biopsies early after treatment using phospho MAPK , Phospho-P70 and Ki67 as end points demonstrated pathway abrogation in the Detroit 562 tumours treated with the combination , the only group where regressions were seen . In conclusion , an P42345 inhibitor showed antitumor activity in P00533 -resistant SCC cell lines . Marked antitumor effects were associated with dual pathway inhibition , which were detected by early FNA biopsies . First analysis of the relation between P33261 genotype and pharmacodynamics in patients treated with ticagrelor versus clopidogrel : the ONSET/OFFSET and RESPOND genotype studies . BACKGROUND : The influence of cytochrome P450 ( CYP ) 2C19 genotype on platelet function in patients treated with ticagrelor versus clopidogrel is unknown . METHODS AND RESULTS : P33261 ( *1 , *2 , *3 , *4 , *5 , *6 , *7 , *8 , *17 ) genotyping was performed in patients with coronary artery disease treated with ticagrelor ( 180-mg load , 90 mg P55957 ) ( n=92 ) or clopidogrel ( 600-mg load , 75 mg/d ) ( n=82 ) . All patients received 75 to 100 mg/d aspirin . Platelet function was measured by aggregometry , VerifyNow Q9H244 assay , and vasodilator-stimulated phosphoprotein-phosphorylation assay at predose , 8 hours postloading , and maintenance . In each treatment group , patients were categorized according to 2C19 genotype carrier status ( loss-of-function , gain-of-function ) and metabolizer status . Kruskal-Wallis test was used to compare platelet function among these categories for each treatment , and Wilcoxon rank sum test was used to compare platelet function between the clopidogrel and ticagrelor groups for each category . There was no statistically significant influence of genotype on platelet function during aspirin therapy alone . DB08816 exhibited lower platelet reactivity than clopidogrel by all assays irrespective of 2C19 genotype or metabolizer status ( P < 0.01 ) . Loss-of-function carriers had greater platelet reactivity during clopidogrel therapy . The influence of genotype on platelet reactivity was greatest during clopidogrel maintenance and best demonstrated by the VerifyNow Q9H244 assay . CONCLUSIONS : This report is the first to demonstrate the superior pharmacodynamic effect of ticagrelor compared with clopidogrel irrespective of P33261 genotype . Whereas P33261 genotype influenced the antiplatelet effect of clopidogrel , there was no effect of P33261 genotype during ticagrelor therapy . 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 . Impact of human D398N single nucleotide polymorphism on intracellular calcium response mediated by α3β4α5 nicotinic acetylcholine receptors . The human P30532 D398N polymorphism ( rs16969968 ) causes an aspartic acid to asparagine change in the nicotinic acetylcholine receptor ( nAChR ) α5 subunit gene . The N398 variant of P30532 is linked to increased risk for nicotine dependence . In this study , we explored the effect of the P30532 D398N polymorphism on the properties of human α3β4* nicotinic acetylcholine receptors in human embryonic kidney ( P29320 ) cells . Addition of either D398 or N398 variant of α5 subunit in the α3β4* receptor did not affect total [ (125)I ] -epibatidine binding or surface expression of the receptor . However , addition of α5(D398) into α3β4* receptor decreased the maximal response to agonist without significantly affecting EC(50) in aequorin intracellular calcium assay . α3β4α5(N398) nAChRs showed further decreased maximal response . The differences in agonist efficacy between the receptor subtypes were found to be dependent upon the concentration of external calcium but independent of external sodium . Moreover , activation of α3β4α5 nAChRs led to significantly greater intracellular calcium release from IP(3) stores relative to α3β4 nAChRs although no effect of the α5 polymorphism was observed . Finally , inclusion of the α5 variant caused a small shift to the left in IC(50) for some of the antagonists tested , depending upon α5 variant but did not affect sensitivity of α3β4* receptors to desensitization in response to incubation with nicotine . In conclusion , addition of either variant of α5 into an α3β4α5 receptor similarly effects receptor pharmacology and function . However , the N398 variant exhibits a reduced response to agonists when extracellular calcium is high and it may lead to distinct downstream cellular signaling . Chromosome 8p as a potential hub for developmental neuropsychiatric disorders : implications for schizophrenia , autism and cancer . Defects in genetic and developmental processes are thought to contribute susceptibility to autism and schizophrenia . Presumably , owing to etiological complexity identifying susceptibility genes and abnormalities in the development has been difficult . However , the importance of genes within chromosomal 8p region for neuropsychiatric disorders and cancer is well established . There are 484 annotated genes located on 8p ; many are most likely oncogenes and tumor-suppressor genes . Molecular genetics and developmental studies have identified 21 genes in this region ( ADRA1A , O15013 , Q15822 , Q15825 , Q05901 , Q9UBT3 , Q16555 , Q06889 , O60258 , Q9NP95 , P11362 , FZD3 , LDL , NAT2 , P07197 , Q02297 , PCM1 , P00750 , P48454 , Q8N474 and P54219 / P54219 ) that are most likely to contribute to neuropsychiatric disorders ( schizophrenia , autism , bipolar disorder and depression ) , neurodegenerative disorders ( Parkinson 's and Alzheimer 's disease ) and cancer . Furthermore , at least seven nonprotein-coding RNAs ( microRNAs ) are located at 8p . Structural variants on 8p , such as copy number variants , microdeletions or microduplications , might also contribute to autism , schizophrenia and other human diseases including cancer . In this review , we consider the current state of evidence from cytogenetic , linkage , association , gene expression and endophenotyping studies for the role of these 8p genes in neuropsychiatric disease . We also describe how a mutation in an 8p gene ( Fgf17 ) results in a mouse with deficits in specific components of social behavior and a reduction in its dorsomedial prefrontal cortex . We finish by discussing the biological connections of 8p with respect to neuropsychiatric disorders and cancer , despite the shortcomings of this evidence . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Role of glutamate and its receptors and insulin-like growth factors in hypoxia induced periventricular white matter injury . This study investigated the glutamate concentration and cellular localization of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid glutamate receptors ( AMPA GluR2 , GluR3 , P48058 ) along with insulin-like growth factors ( IGF ) -1 and -2 expression in the periventricular white matter ( PWM ) of neonatal rats with the aim to determine their involvement in PWM injury in hypoxia . In response to hypoxia , the PWM tissue concentration of glutamate and DB01277 as well as mRNA and protein expression of GluR2 , GluR3 , P48058 , DB01277 , and -2 was upregulated . Immunoexpression of GluR2/3 and P48058 were localized in the amoeboid microglial cells ( AMC ) and oligodendrocytes while that of DB01277 and -2 were confined to AMC . In primary microglial cultures subjected to hypoxia , administration of exogenous glutamate decreased DB01277 but increased the release of tumor necrosis factor-alpha ( P01375 ) and interleukin-1beta ( IL-1beta ) by the cells . Furthermore , silencing of the DB01277 and -2 genes by RNA interference in primary microglial cultures and BV-2 cells downregulated the expression of these growth factors whereas production of glutamate , P01375 , and IL-1beta in these cells was upregulated . It is suggested that increased DB01277 and -2 expressions may be an early protective mechanism in attenuating the hypoxic damage in PWM but a subsequent glutamate-induced decrease of these growth factors may cause cellular injury due to excitotoxicity and increased production of inflammatory cytokines . In this connection , melatonin and edaravone were beneficial in enhancing DB01277 and reducing glutamate release . 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 .

MH_train_1136

MH_train_1136

MH_train_1136

interacts_with DB00945?

multiple_choice

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 . 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 . DB00945 analogues as dual cyclooxygenase-2/ P09917 inhibitors : synthesis , nitric oxide release , molecular modeling , and biological evaluation as anti-inflammatory agents . Analogues of aspirin were synthesized through an efficient one-step reaction in which the carboxyl group was replaced by an ethyl ester , and/or the acetoxy group was replaced by an N-substituted sulfonamide ( SO(2)NHOR(2):R(2) =H , Me , CH(2)Ph ) pharmacophore . These analogues were designed for evaluation as dual cyclooxygenase-2 ( P35354 ) and P09917 ( 5- P28300 ) inhibitors . In vitro P23219 / P35354 isozyme inhibition studies identified compounds 11 ( CO(2) H , SO(2)NHOH ) , 12 ( CO(2)H , SO(2)NHOCH(2)Ph ) , and 16 ( CO(2)Et , SO(2)NHOH ) as highly potent and selective P35354 inhibitors ( IC(50) range : 0.07-0.7 μM ) , which exhibited appreciable in vivo anti-inflammatory activity ( ED(50) range : 23.1-31.4 mg kg(-1) ) . Moreover , compounds 11 ( IC(50) =0.2 μM ) and 16 ( IC(50) =0.3 μM ) , with a sulfohydroxamic acid ( SO(2)NHOH ) moiety showed potent 5- P28300 inhibitory activity . Furthermore , the SO(2)NHOH moiety present in compounds 11 and 16 was found to be a good nitric oxide ( NO ) donor upon incubation in phosphate buffer at pH 7.4 . Molecular docking studies in the active binding site of P35354 and 5- P28300 provided complementary theoretical support for the experimental biological structure-activity data acquired . 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 . [ Variable platelet response to aspirin and new therapeutic targets ] . DB00945 is the first-line oral antiplatelet drug to prevent thromboembolic arterial occlusions . DB00945 irreversibly inhibits cyclooxygenase ( P36551 ) 1 involved in the platelet production of thromboxane ( TX ) A(2) , an inducer of vasoconstriction and a platelet activating agent . Recurrent vascular events despite aspirin intake , combined with laboratory evidence of poor antiplatelet effect , suggested what has been called " aspirin resistance " . For clarity 's sake a real aspirin resistance would be the absence of P23219 inhibition due to intrinsic platelet factors ( which has never been reported ) . What has been described is ( expected ) variability . P23219 inhibition can be insufficient to modify TX-dependent platelet behaviour . Other agonists , the production of which does not involve P23219 , can stimulate TX-receptors . The antiplatelet effect of aspirin can be insufficient for pharmacokinetic or pharmacodynamic reasons , the latter being further classified as TX-dependent or not . If platelets are so reactive that responses are more TX-independent than normally , then neither aspirin nor any drugs acting on this pathway can do the job . These mechanisms should be better understood and diagnosed , and this is the prerequisite for the development of newer antiplatelet agents . [ Effects of nonsteroidal anti-inflammatory drug celecoxib on expression of cyclooxygenase-2 ( P35354 ) in ovarian carcinoma cell ] . OBJECTIVE : To explore the effects of nonsteroidal anti-inflammatory drug Celecoxib on the expression of cyclooxygenase-2 ( P35354 ) in the SKOV3 cell line and the xenografted nude mice of ovarian carcinoma . METHODS : The expression of P35354 in the SKOV3 cell was determined by reverse transcription polymerase chain reaction ( RT-PCR ) , flow cytometry ( DB00828 ) , and Western blot analysis . The expression of P35354 in tumor cells was measured with Immunocytochemistry . RESULTS : RT-PCR showed that the expression P35354 mRNA was strongly down-regulated in SKOV3 cells after treatment with Celecoxib or DB00945 . DB00828 and Western blot analysis showed that the protein product of P35354 was strongly decreased by Celecoxib or DB00945 . The Celecoxib was more potential effects than DB00945 . The immunocytochemistry result showed that the expression of P35354 in 10 , 25 , 50 mg/kg x d of Celecoxib were lower obviously than it in the control group in Xenografted nude mice . CONCLUSION : The anticarcinogenic effects of Celecoxib is probably related to the down-regulation of P35354 , and can be explained to both P35354 -dependent and -independent mechanisms . 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 . Nonsteroidal anti-Inflammatory drugs and cardiovascular risk . Nonsteroidal anti-inflammatory drugs ( NSAID ) inhibit cyclooxygenase ( P36551 ) enzymes , which exist in at least two isoforms , P23219 and P35354 . DB00945 and older agents in this class are nonselective inhibitors of both P23219 and P35354 . Newer agents termed " coxibs " are selective inhibitors of P35354 . Among the NSAID , only aspirin has been proven to significantly reduce cardiovascular risk , primarily through inhibition of P23219 -mediated platelet aggregation . It has been suggested that other nonselective agents , especially naproxen , may provide some lesser degree of cardioprotection , but conclusive evidence is lacking . Conversely , there are concerns that the P35354 inhibitors may increase cardiovascular risk . However , mechanisms for this potentially adverse cardiovascular effect are unknown , and it is becoming increasingly clear that our understanding of the role of P35354 in cardiovascular function is incomplete . Some studies have demonstrated a potentially beneficial effect of P35354 on cardiovascular function that could be negated by P35354 inhibition , while other studies have reported improved endothelial function with P35354 inhibitors . Additionally , the impact of combined therapy with aspirin and other P36551 inhibitors is not yet clear . This article will review the studies that have examined these issues . DB00758 and aspirin in cardiovascular medicine : responders or not -- current best available evidence . Dual antiplatelet therapy represents an important advance for patients with established coronary artery disease . It is an important strategy for patients with acute coronary syndromes and those undergoing percutaneous transcatheter coronary interventions . DB00758 effectively inhibits ADP-induced platelet activation and aggregation by selectively and irreversibly blocking the P2Y(12) receptor on the platelet membrane . DB00945 works by irreversibly acetylating the cyclooxygenase ( P23219 ) enzyme , thus suppressing the production of thromboxane A(2) ( TxA(2) ) and inhibiting platelet activation and aggregation . Variable platelet response and potential resistance to therapy has emerged with aspirin and clopidogrel . The definitions of antiplatelet agents variability in responsiveness and nonresponsiveness are discussed . DB00758 and aspirin responsiveness as they are measured in the laboratory by various techniques ( platelet aggregometry and point-of-care assays such as platelet function analyzer [ PFA-100 ] and rapid platelet function assay [ RPFA ] ) are evaluated . The mechanisms responsible for variations in responsiveness to antiplatelet agents such as clinical , cellular and genetic factors are defined . DB00945 and clopidogrel resistance are emerging clinical entities with potentially severe consequences such as myocardial infarction , stroke or death . The therapeutic interventions to deal with nonresponsiveness are reported , although specific recommendations are not clearly established . In the future , routine measurement of platelet function in patients with cardiovascular disease may become the standard of care . Personalized antithrombotic treatment strategies may be determined by ex-vivo measurements that identify critical pathways influencing thrombotic risk in the individual patient . 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 . [ 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 . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . 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 . DB00945 modulates innate inflammatory response and inhibits the entry of Trypanosoma cruzi in mouse peritoneal macrophages . The intracellular protozoan parasite Trypanosoma cruzi causes Chagas disease , a serious disorder that affects millions of people in Latin America . Cell invasion by T. cruzi and its intracellular replication are essential to the parasite 's life cycle and for the development of Chagas disease . Here , we present evidence suggesting the involvement of the host 's cyclooxygenase ( P36551 ) enzyme during T. cruzi invasion . Pharmacological antagonist for P23219 , aspirin ( ASA ) , caused marked inhibition of T. cruzi infection when peritoneal macrophages were pretreated with ASA for 30 min at 37°C before inoculation . This inhibition was associated with increased production of IL-1β and nitric oxide ( NO(∙) ) by macrophages . The treatment of macrophages with either NOS inhibitors or prostaglandin E2 ( DB00917 ) restored the invasive action of T. cruzi in macrophages previously treated with ASA . Lipoxin Q96JZ2 -receptor antagonist Boc2 reversed the inhibitory effect of ASA on trypomastigote invasion . Our results indicate that DB00917 , NO(∙) , and lipoxins are involved in the regulation of anti-T. cruzi activity by macrophages , providing a better understanding of the role of prostaglandins in innate inflammatory response to T. cruzi infection as well as adding a new perspective to specific immune interventions . Effect of NSAIDS and P35354 inhibitors on the incidence and severity of asbestos-induced malignant mesothelioma : evidence from an animal model and a human cohort . OBJECTIVES : Non-steroidal anti-inflammatory drugs ( NSAIDs ) and P35354 inhibitors have been associated with lower incidence rates of some cancers . Because asbestos can cause chronic inflammation at the pleural and peritoneal surfaces we hypothesised that NSAID and P35354 inhibitors would inhibit the development of asbestos-induced mesothelioma . MATERIALS AND METHODS : A murine model of asbestos-induced mesothelioma was used to test this hypothesis by providing the NSAID , aspirin , daily in the feed at 50mg/kg or 250 mg/kg . In a parallel study , the relationship between the use of NSAID and P35354 inhibitors and mesothelioma was investigated in a human cohort of 1738 asbestos exposed people living or working in Wittenoom , Western Australia ( a crocidolite mine site ) . RESULTS : DB00945 did not alter the rate of disease development or increase the length of time that mice survived . DB00945 had a small but significant effect on disease latency ( the time between asbestos exposure and first evidence of disease ; p < 0.05 ) but disease progression was not affected by the continued presence of the drug . In the Wittenoom cohort , individuals who reported use of NSAIDs , P35354 inhibitors or both did not have a lower incidence of mesothelioma ( HR=0.85 ; 95 % CI=0.53-1.37 , p=0.50 ) , ( HR=0.69 ; 95 % CI=0.21-2.30 , p=0.55 ) and ( HR=0.43 ; 95 % CI=0.16-1.13 , p=0.087 ) respectively . CONCLUSION : We conclude that NSAIDs and P35354 inhibitors do not moderate mesothelioma development or progression in a human cohort exposed to asbestos and this result is confirmed in an autochthonous mouse model . 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 . Sites of action for future therapy : an adenosine-dependent mechanism by which aspirin retains its antiinflammatory activity in cyclooxygenase-2 and NFkappaB knockout mice . The antiinflammatory action of aspirin is generally attributed to inhibition of cyclooxygenases 1 and 2 , but additional mechanisms are at work . These include inhibition of NFkappaB translocation to the nucleus and the capacity of aspirin to promote accumulation of adenosine , a potent antiinflammatory autocoid . We tested these hypotheses in the murine air pouch model of acute inflammation in wild type mice and in cyclooxygenase 2 or NFkappaB knockouts . The antiinflammatory effects of aspirin , sodium salicylate and indomethacin did not correlate with inhibition of cyclooxygenase in either group . Indeed , aspirin retained its antiinflammatory properties even in P35354 knockouts . Similarly , aspirin was no less antiinflammatory in mice rendered deficient for NFkappaB ( Q14511 ) than in wild type controls . In contrast , dexamethasone lost its antiinflammatory capacity in NFkappaB knockouts . DB00945 and sodium salicylate dramatically increased concentrations of adenosine in exudates , a property shared with methotrexate and sulfasalazine . Removal of adenosine by adenosine deaminase or specific antagonism of adenosine at A(2)receptors completely reversed the antiinflammatory effects of aspirin and sodium salicylate , but not those of dexamethasone . This adenosine-dependent , antiinflammatory effect of aspirin points to another target of drug development . Crossreacting drugs and chemicals . DB00945 and nonsteroidal antiinflammatory drugs ( NSAIDs ) exert their clinical effect through inhibition of prostaglandin H synthases 1 and 2 , also known as cyclooxygenase . This shared effect of P36551 -inhibition is also the mechanism for shared adverse effects . Much of our understanding of cross-reacting drugs and chemicals with aspirin comes from studying asthmatics with aspirin-exacerbated respiratory disease ( AERD ) . DB00945 exacerbated respiratory disease is characterized by recalcitrant sinusitis/polyposis , asthma and precipitation of asthma after ingestion of aspirin and most NSAIDs . Cross-reactions between ASA and NSAIDs occur with first exposure unlike IgE-mediated allergic drug reactions . Cross-reactions between aspirin and other drugs are dependent upon inhibition of the cyclooxygenase-1 isoenzyme . Desensitization to aspirin will result in cross-desensitization to all NSATDs that inhibit P23219 . Despite reports in the literature , there does not appear to he cross-reactions between food coloring , hydrocortisone succinate and monosodium glutamate in individuals with aspirin exacerbated respiratory disease . The new highly selective cyclooxygenase 2 inhibitors are well tolerated in AERD asthmatics who have not been desensitized to aspirin . Because low-dose ASA exerts a cardioprotective effect by irreversible inhibition of P23219 , AERD patients who are at risk for coronary artery disease should be considered for aspirin desensitization . [ DB00945 ] . Acetylsalicylic acid ( aspirin ) was the first synthetized drug 100 years ago . Whereas other drugs have disappeared a long time ago , aspirin has stood the test of time and is one of the most important drugs at the turn of this century . Initially it was used for its analgesic , antipyretic and antiphlogistic properties . They are due to the acetylation of the cyclooxygenase and hence an inhibition of prostaglandin synthesis . At higher doses aspirin inhibits interleukin expression induced by nuclear factor-kappa B . DB00945 is the prototype of nonsteroidal antiinflammatory drugs , which are currently challenged by the development of specific inhibitors of cyclooxygenase isoform 2 ( P35354 ) , which must , however , first prove their efficacy and safety in clinical trials . The inhibition of platelet aggregation by aspirin has only been recognized in the second half of the century . It is due to an inhibition of thromboxane A2 synthesis because of an irreversible cyclooxygenase blockade in platelets . DB00945 has been found to reduce the incidence and death rate of cardiovascular and cerebrovascular events and is nowadays the cornerstone of any secondary prevention in vascular diseases . Newer antiaggregatory agents such as ticlopidine , clopidogrel or IIb/IIIa-blockers have been developed . Most often they are used in conjunction with aspirin and their place has yet to be defined . New modes of action of aspirin continue to be found . Recent examples are an improvement of endothelial dysfunction or a reduced incidence of colorectal cancers . It is therefore likely that aspirin will continue to be a very useful and cheap drug for a very large population and will meet the interest of researchers for many more decades . 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 . 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 . A charybdotoxin-sensitive , Ca(2+)-activated K+ channel with inward rectifying properties in brain microvascular endothelial cells : properties and activation by endothelins . A charybdotoxin-sensitive , Ca(2+)-activated K+ channel was identified in cultured rat brain capillary endothelial cells by using conventional single-channel recording techniques and 86( P06400 +)-influx and efflux experiments . Channel activity was dependent on the presence of Ca2+ on the cytosolic face of the membrane with a threshold concentration of 100 nM . It was inhibited by charybdotoxin ( IC50 30 nM ) and quinine ( IC50 0.1 mM ) but not by apamin . K(Ca) channels showed unusual inward rectifying properties under asymmetrical ionic conditions . They were activated by endothelin-1 ( EC50 0.7 nM ) and endothelin-3 ( EC50 7-10 nM ) . The actions of endothelins were prevented by BQ-123 ( Ki = 8 nM ) in a competitive fashion , hence suggesting the involvement of an P25101 -receptor subtype . The channel activity was unaffected by cyclic AMP- or cyclic GMP-elevating agents . The possible role of the intermediate conductance , Ca(2+)-activated K+ channels for mediating K+ movements across the blood-brain barrier is discussed . Synthetic triterpenoid induces P15428 expression and suppresses inflammation-driven colon carcinogenesis . Colitis-associated colon cancer ( CAC ) develops as a result of inflammation-induced epithelial transformation , which occurs in response to inflammatory cytokine-dependent downregulation of 15-hydroxyprostaglandin dehydrogenase ( P15428 ) and subsequent suppression of prostaglandin metabolism . Agents that both enhance P15428 expression and suppress cyclooxygenase-2 ( P35354 ) production may more effectively prevent CAC . Synthetic triterpenoids are a class of small molecules that suppress P35354 as well as inflammatory cytokine signaling . Here , we found that administration of the synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-C28-methyl ester ( CDDO-Me ) suppresses CAC in mice . In a spontaneous , inflammation-driven intestinal neoplasia model , deletion of Q13485 specifically in T cells led to progressive production of inflammatory cytokines , including P01375 -α , IFN-γ , P35228 , P05231 , IL-1β ; as well as activation of P42224 and P40763 ; along with suppression of P15428 expression . Oral administration of CDDO-Me to mice with Q13485 -deficient T cells increased survival and suppressed intestinal epithelial neoplasia by decreasing production of inflammatory mediators and increasing expression of P15428 . Induction of P15428 by CDDO-Me was dose dependent in epithelial cells and was abrogated following treatment with TGF-β signaling inhibitors in vitro . Furthermore , CDDO-Me-dependent P15428 induction was not observed in P84022 -/- mice . Similarly , CDDO-Me suppressed azoxymethane plus dextran sodium sulfate-induced carcinogenesis in wild-type animals , highlighting the potential of small molecules of the triterpenoid family as effective agents for the chemoprevention of CAC in humans . 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 . Selective cyclooxygenase-2 inhibitor cross-reactivity in aspirin-exacerbated respiratory disease . DB00945 -induced asthma ( AIA ) is a severe and difficult-to-treat allergic disease in which acute asthma attacks are induced by nonsteroidal anti-inflammatory drugs . Patients with AIA rarely experience asthma attacks when taking celecoxib , a specific inhibitor of cyclooxygenase ( P36551 ) 2 . A 33-year-old woman had a severe asthma attack with hypoxia and lost consciousness after oral provocation testing with 15 mg of aspirin and also with 50 mg of celecoxib . After 2 months of treatment with 10 mg/day of oral prednisolone , 1600 μg/day of inhaled fluticasone propionate , montelukast as a leukotriene receptor antagonist ( LTRA ) , and long-term beta-agonist , we again challenged her with a provocation test with up to 200 mg of celecoxib ; this time there were neither allergic symptoms nor decrease in forced expiratory volume in 1 second . Patients with severe or poorly controlled asthma may experience asthma attacks even if using selective P35354 inhibitors . However , treatment with steroids and an LTRAs may inhibit asthma attacks induced by celecoxib . 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 . 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 . DB00945 inhibits lipopolysaccharide-induced P35354 expression and DB00917 production in porcine alveolar macrophages by modulating protein kinase C and protein tyrosine phosphatase activity . DB00945 has been demonstrated to be effective in inhibiting P35354 and PGE(2) in Alveolar macrophages ( AMs ) . However , the mechanisms have not been fully understood . In the present study , we found that pretreatment with aspirin inhibited LPS-induced P35354 and PGE(2) upregulation , IκBα degradation , NFκB activation and the increase of PKC activity , but elevated LPS-induced the decrease of PTP activity . The PKC inhibitor calphostin C dramatically reduced the P35354 mRNA and PGE(2) levels , but the PTP inhibitor peroxovanadium ( POV ) significantly increased the P35354 mRNA and PGE(2) levels . Furthermore , the PTP inhibitor mitigated the inhibitory effect of aspirin on P35354 and PGE(2) upregulation and NF-κB activation , whereas the PKC inhibitor enhanced the inhibitory effects of aspirin on the production of P35354 and PGE(2) . Our data indicate a novel mechanism by which aspirin acts as a potent anti-inflammatory agent in alveolus macrophages and ALI . 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 . 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 . 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 . Expression of retinoic acid receptor beta in dermatofibrosarcoma protuberans . BACKGROUND : P10826 ( RAR beta ) has been shown to act as a tumor suppressor in many solid human tumors . To investigate the putative role of RAR beta in dermatofibrosarcoma protuberans ( DFSP ) , we examined the expression of RAR beta in DFSPs and analyzed the correlation of expression patterns between RAR beta and cyclooxygenase ( P36551 ) -2 as well as clinicopathological variables . METHODS : Using tissue microarray and immunohistochemistry , we evaluated nuclear RAR beta staining and cytoplasm P35354 staining in 53 DFSPs . RESULTS : 48 DFSPs ( 90.58 % ) were immunopositive for RAR beta , while 32 DFSPs ( 60.38 % ) were immunopositive for P35354 . RAR beta staining was significantly inversely correlated with P35354 staining ( p < 0.001 ; r =-0.668 ) . CONCLUSIONS : Our data indicated that RAR beta expressed in DFSPs and correlated with P35354 expression . RAR beta may be a potential therapeutic target for unresectable DFSP cases . The effect of aspirin and nonsteroidal anti-inflammatory drugs on prostaglandins . Cyclic prostanoids play important physiologic roles in inflammation and maintaining normal function of several organ systems . Prostaglandin production requires the conversion of arachidonate to the intermediate prostaglandin H2 , by the 2-step cyclo-oxygenation and peroxidation catalyzed by the enzyme cyclo-oxygenase ( also called prostaglandin H synthase ) . DB00945 and nonsteroidal anti-inflammatory drugs ( NSAIDs ) block the production of cyclic prostanoids by binding in different ways to this enzyme and blocking the active site . This results in decreased inflammation , but it can also produce side effects in the gastrointestinal tract , kidney , and platelets . Recent data demonstrate that there are 2 isoforms of the cyclo-oxygenase enzyme , called P23219 and P35354 . These isoforms are similar in size , substrate specificity , and kinetics , but vary in their expression and distribution . Normal physiologic functions appear to be maintained by P23219 , while P35354 appears to mediate the inflammatory response . Nonsteroidal drugs with selective inhibitory activity on the P35354 isoform should theoretically decrease inflammation while maintaining normal physiologic prostaglandin levels . Current NSAIDs are not selective enough to confirm this , but newer , more selective inhibitors of P35354 may answer this important question . DB00945 " resistance " . A variable responsiveness to antiplatelet drugs is a clinical phenomenon that does not principally differ from other drug treatments in other therapeutic fields . The pharmacological part is to clarify whether a " true " resistance exists in pharmacological terms , i.e. , a reduced potency of the compound to work as suggested and to find out the underlying cellular mechanism(s) . Two principally different methods of laboratory control for platelet sensitivity to aspirin ( ASA ) are available : measurement of platelet function ( ex vivo ) or measurement of inhibition of thromboxane formation . Both methods have limitations and did not yet result in a generally accepted definition of a pharmacological ASA " resistance " . The new typological approach of Weber et al . [ A.A. Weber , B. Przytulski , A. Schanz , et al. , Towards a definition of aspirin resistance : a typological approach. Platelets 13 ( 2002 ) 37. ] helps to identify different subtypes of ASA resistance in pharmacological terms by combining in vitro aggregometry with thromboxane measurement . Using this method , a " true " pharmacological resistance , associated with a reduced antiplatelet response to ASA and reduced inhibition of thromboxane formation , was found in patients undergoing coronary artery bypass surgery . Platelets of these patients expressed a hitherto unknown isoform of P35354 - P36551 -2a which might generate a different gene product . In this context , it is interesting that CABG patients express transiently an immunoreactive P35354 protein with lower molecular weight . Studies on the significance of this finding for ASA resistance are in progress . 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 . Interferon-gamma and interleukin 4 inhibit interleukin 1beta-induced delayed prostaglandin E(2)generation through suppression of cyclooxygenase-2 expression in human fibroblasts . Interleukin (IL-)1 stimulates prostaglandin E(2)(PGE(2)) generation in fibroblasts , and preferential couplings between particular phospholipase A(2)(PLA(2)) and cyclooxygenase ( P36551 ) isozymes are implicated with IL-1-induced delayed PGE(2)generation . The regulatory effects of interferon ( IFN ) -gamma and P05112 on IL-1beta-induced P36551 , PLA(2)isoforms expression and terminal delayed PGE(2)generation were examined in three types of human fibroblasts . These human fibroblasts constitutively expressed cytosolic PLA(2)(cPLA(2)) and P23219 enzymes , and exhibited delayed PGE(2)generation in response to IL-1beta . IL-1beta also stimulated expression of cPLA(2)and P35354 only , while constitutive and IL-1beta-induced type IIA and type V secretory PLA(2)s ( sPLA(2)s ) expression could not be detected . A P35354 inhibitor and cPLA(2)inhibitor markedly suppressed the IL-1beta-induced delayed PGE(2)generation , while a type IIA sPLA(2)inhibitor failed to affect it . P01579 and P05112 dramatically inhibited the IL-1beta-induced delayed PGE(2)generation ; these cytokines apparently suppressed IL-1beta-stimulated P35354 expression and only weakly suppressed cPLA(2)expression in response to IL-1beta . These results indicate that IL-1beta-induced delayed PGE(2)generation in these human fibroblasts mainly depends on de novo induction of P35354 and cPLA(2) , irrespective of the constitutive presence of P23219 , and that P01579 and P05112 inhibit IL-1beta-induced delayed PGE(2)generation by suppressing , predominantly , P35354 expression . P35354 and P00533 : Partners in Crime Split by DB00945 . 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 . DB00945 and sodium salicylate inhibit endothelin P25101 receptors by an allosteric type of mechanism . DB00945 is a commonly used drug with a wide pharmacological spectrum including antiplatelet , anti-inflammatory , and neuroprotective actions . This study shows that aspirin and sodium salicylate , its major blood metabolite , reverse contractile actions of endothelin-1 ( ET-1 ) in isolated rat aorta and human mammary arteries . They also prevent the intracellular Ca(2+) mobilizing action of ET-1 in cultured endothelial cells but not those of neuromedin B or UTP . Inhibition of the actions of ET-1 by salicylates is apparently competitive . Salicylates inhibit (125)I-ET-1 binding to recombinant rat P25101 receptors . Salicylic acid promotes dissociation of (125)I-ET-1 P25101 receptor complexes both in the absence and the presence of unlabeled ET-1 . It has no influence on the rate of association of (125)I-ET-1 to P25101 receptors . Salicylates do not promote dissociation of (125)I-ET-1 ETB receptor complexes . Salicylates potentiate relaxing actions of receptor antagonists such as DB00559 . It is concluded that salicylates are allosteric inhibitors of P25101 receptors . The results also suggest that : 1 ) irreversible ET-1 binding probably limits actions of receptor antagonists in vivo , and 2 ) an association of salicylates and P25101 receptor antagonists should be used to evaluate the physiopathological role of ET-1 and may be of therapeutic interest in the treatment of ischemic heart disease . 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 . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . The role of cyclooxygenase in gastric mucosal protection . P23219 and P35354 are two cyclooxygenase enzymes responsible for prostanoid production . P35354 is expressed in inflammatory cells and fibroblasts of the gastric mucosa , and through the production of various growth factors including hepatocyte growth factor ( P14210 ) and vascular endothelial growth factor ( P15692 ) , plays a key role in the tissue repair process . DB00945 induces and acetylates P35354 to produce 15-(R)-epi-lipoxinA4 , an anti-inflammatory mediator thought to protect the gastric mucosa against aspirin-induced injury . Recently , three different PGE synthases have been identified , that convert P35354 metabolites into DB00917 . mPGE synthase ( mPGES ) -1 has been shown to be inducible , and to colocalize with P35354 in fibroblasts and macrophages infiltrating the gastric ulcer bed . Q15185 and Q9H7Z7 have been found expressed in normal gastric mucosa , with no change in expression levels seen in gastritis or gastric ulcer tissue . Finally , this review discusses the role of these enzymes in the pathophysiology of the gastric mucosa , as well as the biologcal significance of their inhibition . Delayed neonatal closure of the ductus arteriosus following early in utero exposure to indomethacin in the rat . BACKGROUND : Indomethacin is used to close the patent ductus arteriosus in premature infants and for tocolysis of preterm labor . Clinically and experimentally , early in utero exposure to indomethacin induces the paradoxical delay of postnatal closure of the ductus arteriosus . OBJECTIVES : To clarify the pharmacological nature of the delay of closure of the ductus arteriosus in the rat . METHODS : We studied early in utero exposure to indomethacin ( dose and timing ) in addition to other drugs , inducing a delay in postnatal ductal closure . Pregnant rats at near term were studied by cesarean section on gestational day 21 ( D21 ) , incubated in room air at 33 degrees C , followed by rapid whole-body freezing . RESULTS : The delay in closure of the ductus arteriosus was dose dependent . A large dose of indomethacin ( 10 mg/kg ) 1 or 2 days before birth induced a delay of 3-4 times . A timing study revealed maximum delay with administration of indomethacin 2 days before birth and minimum delay with administration 5 days before . DB00945 , ibuprofen , the selective P23219 inhibitor SC 560 , the selective P35354 inhibitor rofecoxib and a prostaglandin EP(4) receptor blocker , ONO-208 , all delayed neonatal ductal closure following maternal administration on D19 and D20 . CONCLUSIONS : The delay by indomethacin was dose dependent . The maximum delay was induced by 2 doses of 10 mg/kg indomethacin on D19 and D20 . The delay was induced by a decreased stimulus to the prostaglandin EP(4) receptor system in the last 2 days in utero . The delay was temporary with recovery 3 days or more after exposure . DB00945 may modify tumor microenvironment via antiplatelet effect . High-quality evidence suggests that aspirin is a promising agent for cancer prevention and treatment . Direct inhibition of cyclooxygenase-2 ( P35354 ) pathway is generally thought to be the main mechanism by which aspirin inhibits cancer development . However , either pharmacological properties of aspirin or recent results of epidemiologic studies do not support that mechanism . To address this inconsistency , we hypothesize that antiplatelet effect of aspirin via inhibition of P23219 may be one of potential mechanisms to inhibit carcinogenesis . Aberrant platelet activation will lead to promote hostility of tumor microenvironment by releasing an abundant array of angiogenesis regulators . Given the outstanding ability of antiplatelet , aspirin may restore balance of pro- and anti-angiogenic factors released from platelet to " normalize " tumor vasculature and shape tumor microenvironment to some extent , which will not only diminish tumor aggressiveness and progression , but also enhance the sensitivity to therapeutic treatment . Thus , targeting the platelet activation leading to alter tumor microenvironment may provide a novel way to tumor therapy . Hypothesis : decreased use of pediatric aspirin has contributed to the increasing prevalence of childhood asthma . BACKGROUND : The prevalence of asthma , atopic eczema , and allergic rhinitis has increased over the last three decades in Western countries . Speculation on the causes of this trend have focused on changes in environmental factors . We hypothesize that the decreased use of aspirin in favor of acetaminophen , due to the association of aspirin with Reye 's syndrome during febrile respiratory infections , may be contributing to these trends in the United States . DATA SOURCES : A detailed literature search was conducted utilizing Medline . Studies considered relevant and important involving both humans and animals in English language were used . HYPOTHESIS : In the United States , the documented prevalence of childhood asthma has increased since 1970 , but the rate of this increase accelerated upward beginning in the early 1980s when the use of pediatric aspirin decreased . During the resolution of common respiratory viral infections , prostaglandin E2 ( DB00917 ) is produced through the actions of cyclooxygenase-2 ( P35354 ) . DB00945 , but not acetaminophen , inhibits P35354 activity . As DB00917 promotes TH2 and inhibits THI type cytokine generation , we hypothesize that the decreased use of aspirin may be a factor in facilitating allergic sensitization and asthma by augmenting the relative Q8IXH7 /TH2 cytokine imbalance in genetically predisposed children . CONCLUSION : We have presented an hypothesis based upon epidemiologic trends , known biologic effects of cytokines and DB00917 on allergic sensitization , and a potentially relevant pharmacologic effect of aspirin to explain a component of the increasing prevalence of childhood asthma in the United States . We suggest this theory be examined further in animal models as well as in other countries where the prevalence of childhood asthma is increasing . Protein kinase Cdelta and calmodulin regulate epidermal growth factor receptor recycling from early endosomes through Arp2/3 complex and cortactin . The intracellular trafficking of the epidermal growth factor receptor ( P00533 ) is regulated by a cross-talk between calmodulin ( P62158 ) and protein kinase Cdelta ( PKCdelta ) . On inhibition of P62158 , PKCdelta promotes the formation of enlarged early endosomes and blocks P00533 recycling and degradation . Here , we show that PKCdelta impairs P00533 trafficking due to the formation of an F-actin coat surrounding early endosomes . The PKCdelta-induced polymerization of actin is orchestrated by the Arp2/3 complex and requires the interaction of cortactin with PKCdelta . Accordingly , inhibition of actin polymerization by using cytochalasin D or by overexpression of active cofilin , restored the normal morphology of the organelle and the recycling of P00533 . Similar results were obtained after down-regulation of cortactin and the sequestration of the Arp2/3 complex . Furthermore we demonstrate an interaction of cortactin with P62158 and PKCdelta , the latter being dependent on P62158 inhibition . In summary , this study provides the first evidence that P62158 and PKCdelta organize actin dynamics in the early endosomal compartment , thereby regulating the intracellular trafficking of P00533 . Angioedema associated with aspirin and rofecoxib . OBJECTIVE : To report the probable association of angioedema with aspirin therapy and the selective cyclooxygenase-2 ( P35354 ) inhibitor rofecoxib . CASE SUMMARY : A 44-year-old white woman , previously tolerant to aspirin and other nonsteroidal antiinflammatory drugs ( NSAIDs ) , developed angioedema of the lips after ingesting two 325-mg aspirin tablets during one day . The reaction occurred 3 hours after taking the second aspirin and resolved within 3 hours . Two weeks later , the patient took a 25-mg rofecoxib tablet for a sore throat , and she developed angioedema 5(1/2) hours later . Although the woman took 50 mg of diphenhydramine , the swelling did not subside . She repeated the diphenhydramine dose in the evening and , by noon the next day , 26(1/2) hours after the angioedema began , it was resolved . The patient 's internist prescribed an epinephrine auto-injector and advised her to consult an allergist . With skin testing and oral rechallenge with aspirin , but not rofecoxib , the allergist determined the cause of the reactions to be aspirin-induced angioedema and selective P35354 inhibitor intolerance . The Naranjo probability scale indicated that aspirin was a highly probable cause and rofecoxib was a probable cause of this patient 's angioedema . DISCUSSION : DB00945 -induced angioedema and NSAID intolerance have been well documented . There are reports of both tolerance and intolerance to selective P35354 inhibitors in patients with documented allergy-like reactions to aspirin and NSAIDs . CONCLUSIONS : Patients with aspirin and NSAID intolerance may develop intolerance to P35354 inhibitors , especially with repeated exposure . 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 . 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 . 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 . 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 . Role of interleukin 1 in the regulation of cyclooxygenase gene expression in rat endometrial stromal cells . Interleukin 1 alpha ( P01583 ) stimulates prostaglandin production and cyclooxygenase activity in endometrial stromal cells isolated from the uteri of ovariectomized rats that have been sensitized for the decidual cell reaction . The aim of the present study was to examine the effect of P01583 on the amount of cyclooxygenase mRNA and protein in these cells . Treatment with P01583 ( 20 ng ml-1 ) for 24 h significantly increased steady-state concentrations of cyclooxygenase 2 ( P35354 ) mRNA and protein in the cells , as determined by northern and western blot analyses , respectively . Cyclooxygenase 1 ( P23219 ) mRNA and protein were not detected . Dexamethasone ( 5 mumol l-1 ) prevented the P01583 -induced increase in P35354 steady-state mRNA . Immunocytochemical staining of P35354 in the treated cells indicated that P01583 increased staining , while dexamethasone inhibited this increase . Furthermore , the changes in staining were generalized and not confined to a small subpopulation of cells . These data demonstrate that P01583 increases steady-state concentrations of P35354 mRNA and protein in endometrial stromal cells isolated from the uteri of rats that have been sensitized for decidualization . 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 . 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 . DB00945 , ibuprofen , and other non-steroidal anti-inflammatory drugs in cancer prevention : a critical review of non-selective P35354 blockade ( review ) .

MH_train_1137

MH_train_1137

MH_train_1137

interacts_with DB01045?

multiple_choice

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 . Genetic variation in the retinoid X receptor and calcium-sensing receptor and risk of colorectal cancer in the Colon Cancer Family Registry . Genetic variants in the calcium/vitamin D metabolic pathway may be related to risk for colorectal cancer . While several investigations of vitamin D receptor ( P11473 ) polymorphisms and colorectal cancer have been conducted , no studies to date have evaluated the association of genetic variation in the heterodimer partner for P11473 , the retinoid X receptor ( RXR ) . Another important gene in this pathway is the calcium-sensing receptor ( P41180 ) . Employing a discordant-sibship case-control design , we examined the association between single nucleotide polymorphisms ( SNPs ) in P19793 and P41180 and risk for colorectal cancer overall and by colorectal subsite and microsatellite instability ( MSI ) status using data from the Colon Cancer Family Registry . No gene-level relationships between P19793 or P41180 and colorectal cancer overall were observed . However , for P19793 SNP rs7861779 , a high-interest SNP selected for study a priori , there was a statistically significantly increased risk for proximal colorectal cancer among those with at least one A allele [ odds ratio ( OR ) = 1.42 ; 95 % confidence interval ( CI ) = 1.03-1.97 ] . Another selected P19793 SNP , rs12004589 , was significantly associated with risk of MSI-high cancers ( OR = 2.27 ; 95 % CI = 1.13-4.56 ) . Additionally , P41180 SNP rs1801726 was significantly associated with a reduced risk for rectal cancer ( OR = 0.53 ; 95 % CI = 0.29-0.96 ) . These results provide support that P19793 SNPs rs7861779 and rs12004589 and P41180 SNP rs1801726 may be important markers for colorectal neoplasia . Further work is needed to elucidate their role in the carcinogenic pathway . 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 . 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 . 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 . Effects of low molecular weight heparin on a severely antithrombin III-decreased disseminated intravascular coagulation model in rabbits . The effect of dalteparin , a low molecular weight heparin , on severely antithrombin III ( P01008 ) -decreased disseminated intravascular coagulation ( DIC ) model was compared with that of unfractionated heparin ( heparin ) . The DIC model in rabbits was produced by continuous infusion of thrombin in combination with bolus injection of latex . After a 3 hr infusion of thrombin , plasma P01008 activity was lowered to 30 % of normal plasma . Platelet number , fibrinogen content and alpha 2 plasmin inhibitor ( alpha 2PI ) activity were also decreased . DB06779 ( 25-100 IU/kg/hr ) and heparin ( 25-100 U/kg/hr ) inhibited the decrease in P01008 activity , platelet number and fibrinogen content , and had no effect on alpha 2PI activity . Activated partial thromboplastin time ( APTT ) was prolonged by heparin ( 50 and 100 U/kg/hr ) , but not by dalteparin ( 25-100 IU/kg/hr ) . The ratio of anti-factor Xa ( F.Xa ) activity to anti-thrombin activity for dalteparin ( 50 IU/kg/hr ) was higher than that for heparin ( 50 U/kg/hr ) . With the addition of exogenous P01008 , the ratio of anti-F.Xa to anti-thrombin for heparin increased , but that for dalteparin did not change . However , the increased ratio for heparin was still lower than the unchanged ratio for dalteparin . These results suggest that both dalteparin and heparin have the ability to rectify the abnormal parameters of severely P01008 -decreased DIC , and that the effects of dalteparin are mainly involved with anti-F.Xa activity whereas the effects of heparin are via anti-thrombin activity . 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 . 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 . 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 . 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 . miR-33 controls the expression of biliary transporters , and mediates statin- and diet-induced hepatotoxicity . Bile secretion is essential for whole body sterol homeostasis . Loss-of-function mutations in specific canalicular transporters in the hepatocyte disrupt bile flow and result in cholestasis . We show that two of these transporters , O95342 and O43520 , are functional targets of miR-33 , a micro-RNA that is expressed from within an intron of Q12772 . Consequently , manipulation of miR-33 levels in vivo with adenovirus or with antisense oligonucleotides results in changes in bile secretion and bile recovery from the gallbladder . Using radiolabelled cholesterol , we show that systemic silencing of miR-33 leads to increased sterols in bile and enhanced reverse cholesterol transport in vivo . Finally , we report that simvastatin causes , in a dose-dependent manner , profound hepatotoxicity and lethality in mice fed a lithogenic diet . These latter results are reminiscent of the recurrent cholestasis found in some patients prescribed statins . Importantly , pretreatment of mice with anti-miR-33 oligonucleotides rescues the hepatotoxic phenotype . Therefore , we conclude that miR-33 mediates some of the undesired , hepatotoxic effects of statins . Down-regulation of cholesterol 7alpha-hydroxylase ( P22680 ) gene expression by bile acids in primary rat hepatocytes is mediated by the c-Jun N-terminal kinase pathway . DB04540 7alpha-hydroxylase ( P22680 ) , the rate-limiting enzyme in the neutral pathway of bile acid biosynthesis , is feedback-inhibited at the transcriptional level by hydrophobic bile acids . Recent studies show that bile acids are physiological ligands for farnesoid X receptor ( Q96RI1 ) . Activated Q96RI1 indirectly represses P22680 transcription through induction of small heterodimer protein ( Q15466 -1 ) . In this study , we provide evidence that bile acids rapidly down-regulate P22680 transcription via activation of the JNK/c-Jun pathway . Furthermore , we demonstrate that Q15466 -1 is also a direct target of activated c-Jun . In primary rat hepatocyte cultures , taurocholate ( TCA ) strongly activated JNK in a time- and concentration-dependent manner . P01375 -alpha , a potent activator of JNK , also rapidly activated JNK and down-regulated P22680 mRNA levels . Overexpression of dominant-negative P45983 or a transactivating domain mutant of c-Jun significantly blocked the ability of TCA to down-regulate P22680 mRNA . In contrast , overexpression of wild-type c-Jun ( c-Jun(wt) ) enhanced the repression of P22680 by TCA . Moreover , overexpression of c-Jun(wt) resulted in increased Q15466 -1 promoter activity . Mutation of a putative AP-1 ( c-Jun ) element suppressed c-Jun-mediated activation of the Q15466 -1 promoter construct . These results indicate that the bile acid-activated JNK pathway plays a pivotal role in regulating P22680 levels in primary rat hepatocytes . 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 . Monocyte gene expression signature of patients with early onset coronary artery disease . The burden of cardiovascular disease ( CVD ) can not be fully addressed by therapy targeting known pathophysiological pathways . Even with stringent control of all risk factors CVD events are only diminished by half . A number of additional pathways probably play a role in the development of CVD and might serve as novel therapeutic targets . Genome wide expression studies represent a powerful tool to identify such novel pathways . We compared the expression profiles in monocytes from twenty two young male patients with premature familial CAD with those from controls matched for age , sex and smoking status , without a family history of CVD . Since all patients were on statins and aspirin treatment , potentially affecting the expression of genes in monocytes , twelve controls were subsequently treated with simvastatin and aspirin for 6 and 2 weeks , respectively . By whole genome expression arrays six genes were identified to have differential expression in the monocytes of patients versus controls ; O95477 , P45844 and Q08116 were downregulated in patients , whereas P07550 , P41439 and P09488 were upregulated . Differential expression of all genes , apart from P09488 , was confirmed by qPCR . DB00945 and statins altered gene expression of P45844 and ADBR2 . All finding were validated in a second group of twenty four patients and controls . Differential expression of O95477 , Q9BU20 and ADBR2 was replicated . In conclusion , we identified these 3 genes to be expressed differently in CAD cases which might play a role in the pathogenesis of atherosclerotic vascular disease . 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 . Effect of P14416 , 5- Q13049 , and P21964 genes on antipsychotic response to risperidone . DB00734 is a widely used atypical antipsychotic with certain advantages over typical antipsychotics . Although variations in the efficacy of treatment with risperidone have been observed , no specific predictable marker has been identified as of yet . In all , 73 Japanese patients with schizophrenia were given risperidone for 8 weeks , and clinical symptoms were evaluated using the Positive and Negative Syndrome Scale ( PANSS ) . Six candidate polymorphisms ( P28223 -1438G > A , 102T > C , H452Y ; P14416 -141delC , Taq I A ; P21964 V158M ) were genotyped . The diplotype configuration for each individual was estimated by the maximum-likelihood method . Multiple linear regressions were used to analyze the effects of these haplotypes/genotype and other prognostic factors on PANSS scale performance . After adjustment for the effects of patient-related variables , P28223 diplotype and P21964 genotype , as well as other potential prognostic factors , did not significantly influence the clinical performance . A P14416 haplotype tended to correlate with better clinical performance . Compared with patients who had Ins-A2/Ins-A2 diplotype ( n=25 ) , PANSS total scores of patients with Ins-A2/Del-A1 diplotype ( n=10 ) showed 40 % greater improvement ( P=0.03 ) . The PANSS total scores of patients with P28223 A-T/A-T diplotype ( n=22 ) tended to show 15 % worse improvement compared with A-T/G-C diplotype ( n=33 ) ( P=0.06 ) . These results should be treated with caution because of limitations due to small sample size , heterogeneity of patients with respect to past antipsychotic use history , and no correction for multiple corrections . However , the present findings generate important hypotheses in a sample of Japanese schizophrenia patients that may lay the foundation for future pharmacogenomics investigations in other populations . 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 . 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 . 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 . DB01095 enhances the inhibitory effects of a selective AT1 receptor blocker , valsartan , on atherosclerosis . We investigated the effects of a P04035 inhibitor ( statin ) on the inhibitory effects of an angiotensin II type-1 receptor ( AT1 ) blocker on atherosclerosis and explored cellular mechanisms . We gave apolipoprotein E null mice a high-cholesterol diet for 10 weeks and measured atherosclerotic plaque area and lipid deposition . Neither 1 mg/kg per day of valsartan nor 3 mg/kg per day of fluvastatin had any effect on blood pressure or cholesterol concentration ; however , both drugs decreased plaque area and lipid deposition after 10 weeks . We then reduced the doses of both drugs to 0.1 mg/kg per day and 1 mg/kg per day , respectively . At these doses , neither drug had an effect on atherosclerotic lesions . When both drugs were combined at these doses , a significant reduction in atherosclerotic lesions was observed . Similar inhibitory effects of valsartan or fluvastatin on the expressions of nicotinamide-adenine dinucleotide/nicotinamide-adenine dinucleotide phosphate oxidase subunits P13498 and p47phox , production of superoxide anion , the expression of monocyte chemoattractant protein-1 , and intercellular adhesion molecule-1 expression were observed . These results suggest that concomitant AT1 receptor and cholesterol biosynthesis blockade , particularly when given concomitantly , blunts oxidative stress and inflammation independent of blood pressure or cholesterol-related effects . [ 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 . 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 . 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 . 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 . 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 . 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 . 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 . Peritoneal macrophages mediated delivery of chitosan/siRNA nanoparticle to the lesion site in a murine radiation-induced fibrosis model . BACKGROUND : Radiation-induced fibrosis ( Q9HBH0 ) is a dose-limiting complication of cancer radiotherapy and causes serious problems , i.e. restricted tissue flexibility , pain , ulceration or necrosis . Recently , we have successfully treated Q9HBH0 in a mouse model by intraperitoneal administration of chitosan/siRNA nanoparticles directed towards silencing P01375 alpha in local macrophage populations , but the mechanism for the therapeutic effect at the lesion site remains unclear . METHODS : Using the same murine Q9HBH0 model we utilized an optical imaging technique and fluorescence microscopy to investigate the uptake of chitosan/fluorescently labeled siRNA nanoparticles by peritoneal macrophages and their subsequent migration to the inflamed tissue in the Q9HBH0 model . RESULTS : We observed strong accumulation of the fluorescent signal in the lesion site of the irradiated leg up to 24 hours using the optical imaging system . We further confirm by immunohistochemical staining that Cy3 labeled siRNA resides in macrophages of the irradiated leg . CONCLUSION : We provide a proof-of-concept for host macrophage trafficking towards the inflamed region in a murine Q9HBH0 model , which thereby suggests that the chitosan/siRNA nanoparticle may constitute a general treatment for inflammatory diseases using the natural homing potential of macrophages to inflammatory sites . Complex agonist-like properties of DB00947 ( Faslodex ) in human breast cancer cells that predominantly express progesterone receptor-B : implications for treatment resistance . DB00947 ( Faslodex ) , considered a pure anti-estrogen , is approved for treatment of post-menopausal breast cancer patients who fail to respond to tamoxifen therapy . We recently reported that , like mifepristone , DB00947 exhibits anti-progestin activity , blocking the progestin-dependent increase in endogenous vascular endothelial growth factor ( P15692 ) mRNA and protein release . Some anti-progestins have partial agonist-like activity in breast cancer cells expressing high levels of progesterone receptor B ( PRB ) . Our results show that DB00947 can also induce reporter activity from a plasmid containing a simple progestin responsive element ( PRE ) in these cells . Using small interfering RNA , we determined that induction is dependent on the presence of PR , estrogen receptor and Q15788 . Regulation of more complex progestin-responsive promoters was context-dependent ; induction was observed from the MMTV promoter but not from the P15692 promoter . In contrast , DB00947 increased the release of angiogenically active P15692 from cells expressing elevated levels of PRB . This effect was dependent on the phosphatidylinositol-3 kinase and P29323 /MAPK signaling pathways . We hypothesize that these agonist-like properties of DB00947 ( one genomic and one non-genomic ) may contribute to the acquisition of drug resistance , suggesting that both anti-hormonal and anti-angiogenic treatment may be appropriate in these patients . Associations between the P13498 242C/T and the P05164 -463G/A polymorphisms , oxidative stress and cardiovascular disease in chronic kidney disease patients . Genetic variations in the NADPH/ P05164 system in chronic kidney disease ( CKD ) patients might lead to altered activity of these enzymes , and thus to altered risk for oxidative stress ( OS ) and cardiovascular disease ( CVD ) . We evaluated the impact of 242C/T P13498 and -463G/A P05164 polymorphisms on OS and CVD mortality in stage 5 CKD patients starting dialysis . Two hundred and fifty-seven patients were genotyped using Pyrosequencing . Plasmalogen [ dimethylacetal ( P28067 ) 16/C16:0 ] was used as OS marker . CVD was assessed from patient history and clinical symptoms . Prevalence of CVD was higher ( 35 % ) in GG patients ( P05164 ) compared to AG ( 26 % ) and AA ( 0 % ) patients ( p < 0.01 ) . Patients with CC genotype ( P13498 ) had lower levels of P28067 16/C16:0 ( ratio 0.071 +/- 0.003 ) compared to TT patients ( 0.089 +/- 0.006 ; p < 0.05 ) . These patients also had increased CVD mortality compared to CT and TT patients ( chi(2) P98173 ; p < 0.05 ) . We conclude that genetic variations in the NADPH/ P05164 system are associated with OS , presence of CVD and CVD-related mortality in CKD patients . 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 . 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 % . Redundant roles for cJun-N-terminal kinase 1 and 2 in interleukin-1beta-mediated reduction and modification of murine hepatic nuclear retinoid X receptor alpha . BACKGROUND/AIMS : P19793 ( RXRalpha ) , the heterodimeric partner for multiple nuclear receptors ( NRs ) , was shown to be an essential target for inflammation-induced cJun-N-terminal kinase ( JNK ) signaling in vitro . This study aimed to explore the role of hepatic JNK signaling and its effects on nuclear RXRalpha levels downstream of interleukin-1beta ( IL-1beta ) in vivo . METHODS : Effects of IL-1beta on hepatic NR-dependent gene expression , nuclear RXRalpha levels , and roles for individual JNK isoforms were studied in wild-type , Jnk1(-/-) , and Jnk2(-/-) mice and in primary hepatocytes of each genotype . RESULTS : IL-1beta administration showed a time-dependent reduction in expression of the hepatic NR-dependent genes Ntcp , Cyp7a1 , Cyp8b1 , Abcg5 , Mrp2 , and Mrp3 . IL-1beta treatment for 1h activated JNK and resulted in both post-translational modification and reduction of nuclear RXRalpha . In wild-type primary hepatocytes , IL-1beta modified and reduced nuclear RXRalpha levels time dependently , which was prevented by chemical inhibition of JNK as well as by inhibition of proteasomal degradation . Individual absence of either P45983 or P45984 did not significantly influence the reduction or modification of hepatic nuclear RXRalpha by IL-1beta both in vivo and in primary hepatocytes . CONCLUSIONS : Functional redundancy exists for P45983 and P45984 in IL-1beta-mediated alterations of hepatic nuclear RXRalpha levels , stressing the importance of this pathway in mediating the hepatic response to inflammation . 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 . 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 . Cooperation between Q01196 -ETO9a and novel transcriptional partner Q99612 in upregulation of Alox5 in acute myeloid leukemia . Fusion protein Q01196 - Q06455 ( Q01196 - Q06455 , Q01196 - Q06455 ) is expressed as the result of the 8q22;21q22 translocation [ t(8;21) ] , which is one of the most common chromosomal abnormalities found in acute myeloid leukemia . Q01196 - Q06455 is thought to promote leukemia development through the aberrant regulation of Q01196 ( Q01196 ) target genes . Repression of these genes occurs via the recruitment of the corepressors N-COR and Q9Y618 due to their interaction with Q06455 . Mechanisms of Q01196 - Q06455 target gene upregulation remain less well understood . Here we show that Q01196 -ETO9a , the leukemogenic alternatively spliced transcript expressed from t(8;21) , upregulates target gene Alox5 , which is a gene critically required for the promotion of chronic myeloid leukemia development by P11274 - P00519 . Loss of Alox5 expression reduces activity of Q01196 -ETO9a , Q03164 - P42568 and P29590 -RARα in vitro . However , Alox5 is not essential for the induction of leukemia by Q01196 -ETO9a in vivo . Finally , we demonstrate that the upregulation of Alox5 by Q01196 -ETO9a occurs via the C₂H₂ zinc finger transcription factor Q99612 , a protein required for early hematopoiesis and yolk sac development . Furthermore , Q99612 is specifically upregulated by Q01196 - Q06455 in human leukemia cells . This identifies Q99612 as a novel mediator of t(8;21) target gene regulation , providing a new mechanism for Q01196 - Q06455 transcriptional control . [ 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 . Nuclear translocation of Q02750 triggers a complex T cell response through the corepressor silencing mediator of retinoid and thyroid hormone receptor . Q02750 phosphorylates P27361 /2 and regulates T cell generation , differentiation , and function . Q02750 has recently been shown to translocate to the nucleus . Its nuclear function is largely unknown . By studying human P01730 T cells , we demonstrate that a low level of Q02750 is present in the nucleus of P01730 T cells under basal conditions . T cell activation further increases the nuclear translocation of Q02750 . Q02750 interacts with the nuclear receptor corepressor silencing mediator of retinoid and thyroid hormone receptor ( Q9Y618 ) . Q02750 reduces the nuclear level of Q9Y618 in an activation-dependent manner . Q02750 is recruited to the promoter of c-Fos upon TCR stimulation . Conversely , Q9Y618 is bound to the c-Fos promoter under basal conditions and is removed upon TCR stimulation . We examined the role of Q9Y618 in regulation of T cell function . Small interfering RNA-mediated knockdown of Q9Y618 results in a biphasic effect on cytokine production . The production of the cytokines P60568 , P05112 , P22301 , and IFN-γ increases in the early phase ( 8 h ) and then decreases in the late phase ( 48 h ) . The late-phase decrease is associated with inhibition of T cell proliferation . The late-phase inhibition of T cell activation is , in part , mediated by P22301 that is produced in the early phase and , in part , by β-catenin signaling . Thus , we have identified a novel nuclear function of Q02750 . Q02750 triggers a complex pattern of early T cell activation , followed by a late inhibition through its interaction with Q9Y618 . This biphasic dual effect most likely reflects a homeostatic regulation of T cell function by Q02750 . 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 . 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 . 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 . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . Overexpression of cytochrome P450 4F2 in mice increases 20-hydroxyeicosatetraenoic acid production and arterial blood pressure . P78329 ( P78329 ) activity is thought to be a factor in the pathogenesis of hypertension through its bioactive metabolite 20-hydroxyeicosatetraenoic acid ( 20-HETE ) . We previously found that a gain-in-function P78329 variant in a Chinese cohort was associated with elevated urinary 20-HETE and hypertension . To further explore this association we generated a transgenic mouse model expressing P78329 driven by a modified mouse kidney androgen-regulated protein promoter . This heterologous promoter regulated the expression of luciferase and his-tagged P78329 in transfected P29320 293 cells . In the kidney of transgenic mice , P78329 was localized to renal proximal tubule epithelia and was expressed at a higher level than in control mice , leading to increased urinary 20-HETE excretion . Assessment of P78329 activity by an arachidonic acid hydroxylation assay showed that 20-HETE production was significantly higher in kidney microsomes of transgenic mice compared to control mice , as was their systolic blood pressure . There was a positive correlation of blood pressure with urinary 20-HETE levels . Our results show that increased expression of P78329 in mice enhanced 20-HETE production and elevated blood pressure . Q9Y618 -mediated co-shuttling enables export of class IIa HDACs independent of their P62158 kinase phosphorylation sites . The Class IIa histone deacetylases (HDAC)4 and Q9UQL6 play a role in neuronal survival and behavioral adaptation in the CNS . Phosphorylation at 2/3 N-terminal sites promote their nuclear export . We investigated whether non-canonical signaling routes to Class IIa HDAC export exist because of their association with the co-repressor Silencing Mediator Of Retinoic And Thyroid Hormone Receptors ( Q9Y618 ) . We found that , while Q9UQL6 and P56524 mutants lacking their N-terminal phosphorylation sites ( P56524 ( P22033 ) , Q9UQL6 ( P22033 ) ) are constitutively nuclear , co-expression with Q9Y618 renders them exportable by signals that trigger Q9Y618 export , such as synaptic activity , HDAC inhibition , and Brain Derived Neurotrophic Factor ( P23560 ) signaling . We found that Q9Y618 's repression domain 3 ( Q7Z3Z2 ) is critical for co-shuttling of Q9UQL6 ( P22033 ) , consistent with the role for this domain in Class IIa HDAC association . In the context of P23560 signaling , we found that Q9UQL6 (WT) , which was more cytoplasmic than Q9UQL6 ( P22033 ) , accumulated in the nucleus after P23560 treatment . However , co-expression of Q9Y618 blocked P23560 -induced Q9UQL6 (WT) import in a Q7Z3Z2 -dependent manner . In effect , Q9Y618 -mediated Q9UQL6 (WT) export was opposing the P23560 -induced Q9UQL6 nuclear accumulation observed in Q9Y618 's absence . Thus , Q9Y618 's presence may render Class IIa HDACs exportable by a wider range of signals than those which simply promote direct phosphorylation . Suppression of dendritic cell maturation by Trichinella spiralis excretory/secretory products . Evidence from experimental studies indicates that during chronic infections with certain helminth species a regulatory network is induced that can down-modulate not only parasite-induced inflammation but also reduce other immunopathologies such as allergies and autoimmune diseases . The mechanisms however , and the molecules involved in this immunomodulation are unknown . Here , we focus on the effect of Trichinella spiralis excretory/secretory antigens ( TspES ) on the innate immune response by studying the effect of TspES on DC maturation in vitro . Bone marrow-derived DC from BALB/c mice were incubated with TspES either alone or in combination with LPS derived from two different bacteria . As indicators of DC maturation , the cytokine production ( IL-1alpha , P05231 , P22301 , IL-12p70 and P01375 ) and the expression of various surface molecules ( MHC-II , P25942 , P33681 and P42081 ) were measured . Results indicate that while TspES alone did not change the expression of the different surface molecules or the cytokine production , it completely inhibited DC maturation induced by Escherichia coli LPS ( E. coli LPS ) . In contrast , DC maturation induced by LPS from another bacterium , Neisseria meningitidis , was not affected by TspES . These results were confirmed using O00206 / Q9Y6Y9 / P08571 transfected P29320 293 cells . In conclusion , T. spiralis ES antigens lead to suppression of DC maturation but this effect depends on the type of LPS used to activate these cells . Intracellular volume and apparent diffusion constants of perfused cancer cell cultures , as measured by NMR . Diffusion NMR spectroscopy was used to study intracellular volume and apparent water diffusion constants in different cell lines ( DU145 , human prostate cancer ; P01008 , rat prostate cancer ; MCF-7 , human breast cancer ; Q9HBH0 -1 , mouse fibrosacroma ) . The cells were grown on various matrices ( collagen sponge , collagen beads , polystyrene beads ) which enabled continuous growth in perfused high density cell culture suitable for NMR studies . In perfused cell systems , the attenuation of the water signal versus the squared gradient strength was fitted by the sum of two decaying exponentials . For the slowly decaying component the apparent water diffusion constant at 37 degrees C was 0.22 ( +/-0.02 ) x 10(-9) s/m2 for all cell lines at diffusion times > 100 ms . It continuously increased up to 0.47 ( +/-0.05 ) x 10(-9) s/m2 when the diffusion time was decreased to 8 ms , indicating restricted diffusion . No significant effect of the matrices was observed . The fractional volume of the slow component as determined from the biexponential diffusion curve correlated with the relative intracellular volume , as obtained from the cell density in the sample and the cell size as measured by light microscopy . Therefore , this simple NMR approach can be used to determine intracellular volume in perfused cell cultures suitable for NMR studies . Using this information in combination with spectroscopic data , changes in intracellular metabolite concentration can be detected even when the cellular volume is changing during the experiment . The apparent diffusion constant for the fast diffusing component varied with growth matrix , cell density and cell type and also showed the typical characteristics of restricted diffusion ( increase of apparent diffusion constant with time ) . Regulation of human cytochrome P450 4F2 expression by sterol regulatory element-binding protein and lovastatin . This report provides the first evidence that human P450 4F2 ( P78329 ) is induced by statins , which are widely used to treat hypercholesterolemia . Real time PCR and immunoblots indicate that lovastatin treatment increases expression of the endogenous P78329 gene in human primary hepatocytes and HepG2 cells . The effects of lovastatin on gene expression are often mediated through sterol regulatory element-binding proteins ( SREBPs ) . Immunoblots indicate that lovastatin-treated human hepatocytes display increased proteolytic processing of Q12772 . In HepG2 cells , co-administration of a potent suppressor of Q12772 activation , 25-hydroxycholesterol , inhibits P78329 mRNA induction by lovastatin . HepG2 cells transfected with an expression vector for the active nuclear form of SREBP-1a ( nSREBP-1a ) also display elevated endogenous P78329 expression . Luciferase reporters containing the P78329 proximal promoter are transactivated by nSREBPs ( -1a , -1c , and -2 ) or a dominant positive form of the Q12770 ( SCAP ) , which facilitates activation of endogenous SREBPs . DB00227 -induced reporter expression is inhibited by overexpressed Insig-1 , which prevents proteolytic activation of endogenous SREBPs . Electrophoretic mobility shift assays with in vitro translated nSREBP-1a identified two SREBP binding sites at -169/-152 and -109/-92 , relative to the P78329 transcription start site . Mutations in each site abolish SREBP binding . Chromatin immunoprecipitation experiments indicate that more P36956 is associated with the P78329 promoter after overexpression of nSREBP-1a . Transfection studies and mutagenesis indicate that the -109/-92 region is the primary site responsible for the effects of statins . Collectively , these results demonstrate that SREBPs transactivate P78329 transcription and that P78329 induction by statins is mediated by Q12772 . Functional characterization of a novel serotonin receptor ( 5-HTap2 ) expressed in the CNS of Aplysia californica . Serotonin has been shown to be a neuromodulator in the Aplysia californica CNS . The diversity of serotonin actions is due to the existence of several different receptor subtypes . In this study we report the cloning of a full-length cDNA , coding for a novel serotonin receptor ( 5-HTap2 ) . The receptor protein bears the characteristics of G protein-coupled receptors . It shares 68 % and 34 % of its amino acid sequence identity with the 5-HTlym receptor from Lymnaea stagnalis and the mammalian P08908 receptor , respectively . When transfected in P29320 293 cells , 5-HTap2 was negatively coupled to adenylate cyclase . Ligand binding analysis indicated that the order of potencies of various drugs for the inhibition of [3H]LSD binding was : methiothepin > metergoline > 5-CT > PAPP > 5-HT > ketanserin > NAN-190 > 8-OH-DPAT > clozapine . RT-PCR amplification of RNA isolated from different tissues indicated that this receptor is expressed in the CNS and in bag cells . The expression of 5-HTap2 restricted to the CNS suggests an important role for this receptor in the modulation of neuronal functions in Aplysia . Moreover , the high expression of 5-HTap2 in the bag cells , associated with its pharmacological profile , suggests that this receptor may be implicated in modulating the afterdischarge during the egg-laying behavior . A homozygous inactivating calcium-sensing receptor mutation , Pro339Thr , is associated with isolated primary hyperparathyroidism : correlation between location of mutations and severity of hypercalcaemia . BACKGROUND : Inactivating mutations of the calcium-sensing receptor ( P41180 ) , a G-protein-coupled receptor with extracellular ( O95905 ) , transmembrane ( TMD ) and intracellular ( ICD ) domains , cause familial hypocalciuric hypercalcaemia , neonatal severe primary hyperparathyroidism and occasionally primary hyperparathyroidism in adults . OBJECTIVE : To investigate a patient with typical symptomatic primary hyperparathyroidism for P41180 abnormalities . PATIENT AND DESIGN : A 51-year-old woman with primary hyperparathyroidism was investigated for P41180 abnormalities as her severe hypercalcaemia ( 3·75 mm ) persisted after the removal of two large parathyroid adenomas and she was the daughter of normocalcaemic consanguineous parents . Following informed consent , P41180 mutational analysis was undertaken using leucocyte DNA . Wild-type and mutant P41180 constructs were expressed in human embryonic kidney ( P29320 ) 293 cells and assessed by measuring their intracellular calcium responses to changes in extracellular calcium . Clinical data were pooled with previous studies to search for genotype-phenotype correlations . RESULTS : The proband was homozygous for a Pro339Thr P41180 missense mutation , located in the O95905 , and her normocalcaemic relatives were heterozygous . The mutant Thr339 P41180 had a rightward shift in its dose-response curve with a significantly higher EC(50) = 3·18 mm ± 0·19 compared to the wild-type EC(50) = 2·16 mm ± 0·1 ( P < 0·01 ) , consistent with a loss-of-function mutation . An analysis of P41180 mutations in patients with primary hyperparathyroidism revealed that those of the O95905 were associated with a significantly greater hypercalcaemia that was less likely to be corrected after removal of the parathyroid tumours . CONCLUSIONS : A P41180 missense mutation causing a loss-of-receptor-function can cause symptomatic primary hyperparathyroidism in adulthood . 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 . 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 . 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 . An O95477 truncation shows no dominant negative effect in a familial hypoalphalipoproteinemia pedigree with three O95477 mutations . The DB00171 binding cassette transporter ( O95477 ) A1 is a key determinant of circulating high density lipoprotein cholesterol ( HDL-C ) levels . Mutations in O95477 are a major genetic contributor to low HDL-C levels within the general population . Following the finding of three different O95477 mutations , p.C978fsX988 , p.T1512M and p.N1800H in a subject with hypoalphalipoproteinemia , we aimed to establish whether the p.C978fsX988 truncation exerted a dominant negative effect on the full-length O95477 alleles within family members as has been reported for other O95477 truncations . Characterisation of the p.C978fsX988 mutant in transfected P29320 293 cells showed it to be expressed as a GFP fusion protein but lacking in cholesterol efflux function . This was in keeping with results from cholesterol efflux assays in the fibroblasts of p.C978fsX988 carriers which also showed impaired efflux . Allele- specific quantification of p.C978fsX988 mRNA and analysis of O95477 protein levels in the fibroblasts of p.C978fsX988 heterozygotes showed negligible levels of mRNA and protein expression . There was no evidence of a dominant negative effect on wildtype or p.N1800H protein levels . We conclude that in the case of the p.C978fsX988 truncated mutant a lack of expression precludes it from having a dominant negative effect . A new role for a metabolic star : AMP-activated protein kinase stimulates fat absorption . AMPK is a kinase involved in cell energy homeostasis . In this issue of Cell Metabolism , Chopra and colleagues ( 2011 ) reveal a new function of AMPK in stimulating fat absorption . AMPK enhances the expression of the bile acid transporter , O95342 , by activating the coactivator P12931 -2 , thus promoting bile acid secretion . [ Development of simplified and rapid detection assay for genetic polymorphisms influencing drug response and its clinical applications ] . Clinically important genetic polymorphisms influencing drug metabolism and drug response have typically been discovered on the basis of phenotypic differences among individuals from different populations . Routine genotyping before drug therapy may enable the identification of responders , nonresponders , or patients at increased risk of toxicity . Automated , high-throughput detecting methods for single-nucleotide polymorphisms ( SNPs ) are highly desirable in many clinical laboratories . The aim of this study is to develop a high-throughput genotyping method for detecting SNPs influencing drug response in the Japanese population . We have developed three real-time PCR assays for detecting SNPs in the human drug-metabolizing enzymes and drug targets . The assay for simultaneously detecting P11509 , P20813 , P11712 , P33260 , P33261 , P10635 , P05181 , P20815 , NAT2 , P51580 , Q12882 , P22309 , P05091 , P00325 , P08183 , P11597 , P12821 -1 , P07550 , P28223 , P49441 , P48061 , and mitochondrial DNA polymorphisms takes less than 1.5 h . With the clinical application of NAT2 genotyping , we found statistically significant difference between the incidence of adverse drug reactions ( ADRs ) and the NAT2 genotype . The incidence of the ADRs was significantly higher in the slow type than the in other two types , as 5 of the 6 patients were of the slowtype , and the other was the intermediatetype , while no patients of the rapidtype has developed any ADRs . 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 . 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 .

MH_train_1138

MH_train_1138

MH_train_1138

interacts_with DB00898?

multiple_choice

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 . Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking . Binge drinking of alcoholic beverages among adolescents is a common practice that can have serious health consequences . DB00898 is a potent immunomodulator that alters a wide range of immune responses . However , it is unclear whether there is a differential immune response to alcoholic beverages with a high versus low concentration of ethanol . In this study , we used a PCR array containing 46 primer pairs of selected genes to compare mRNA expression in the spleen , an immune system organ , of adolescent rats following binge drinking of alcohol solutions containing either 20 % or 52 % ethanol ( v/v , 4.8 g/kg daily dosage ) , or water ( control ) for 3 d . We found that , expression of IL-1β , P05231 , P13500 , and GABA(A) receptor α2 subunit in the spleen were decreased , and P41594 and P46098 receptor expression were increased after administration of an ethanol solution containing 52 % ethanol , but not one with 20 % ethanol . Our data suggest that alcohol-mediated immunomodulatory effects are , in part , dependent on the ethanol by volume concentration . This is the first study to show that exposure to a high ethanol percentage beverage can have more profound effects on immune responses than one with a low percentage of ethanol . 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 . Potential genetic risk factors for chronic TMD : genetic associations from the OPPERA case control study . Genetic factors play a role in the etiology of persistent pain conditions , putatively by modulating underlying processes such as nociceptive sensitivity , psychological well-being , inflammation , and autonomic response . However , to date , only a few genes have been associated with temporomandibular disorders ( TMD ) . This study evaluated 358 genes involved in pain processes , comparing allelic frequencies between 166 cases with chronic TMD and 1,442 controls enrolled in the OPPERA ( Orofacial Pain : Prospective Evaluation and Risk Assessment ) study cooperative agreement . To enhance statistical power , 182 TMD cases and 170 controls from a similar study were included in the analysis . Genotyping was performed using the Pain Research Panel , an Affymetrix gene chip representing 3,295 single nucleotide polymorphisms , including ancestry-informative markers that were used to adjust for population stratification . Adjusted associations between genetic markers and TMD case status were evaluated using logistic regression . The OPPERA findings provided evidence supporting previously reported associations between TMD and 2 genes : P28223 and P21964 . Other genes were revealed as potential new genetic risk factors for TMD , including P04150 , Q16566 , P08172 , O00458 , and P34947 . While these findings need to be replicated in independent cohorts , the genes potentially represent important markers of risk for TMD , and they identify potential targets for therapeutic intervention . PERSPECTIVE : Genetic risk factors for TMD pain were explored in the case-control component of the OPPERA cooperative agreement , a large population-based prospective cohort study . Over 350 candidate pain genes were assessed using a candidate gene panel , with several genes displaying preliminary evidence for association with TMD status . 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 . A novel mechanism of autophagic cell death in dystrophic muscle regulated by Q99572 receptor large-pore formation and HSP90 . Q99572 is an DB00171 -gated ion channel , which can also exhibit an open state with a considerably wider permeation . However , the functional significance of the movement of molecules through the large pore ( LP ) and the intracellular signaling events involved are not known . Here , analyzing the consequences of Q99572 activation in primary myoblasts and myotubes from the Dmd(mdx) mouse model of Duchenne muscular dystrophy , we found DB00171 -induced Q99572 -dependent autophagic flux , leading to P42574 - P55210 -independent cell death . Q99572 -evoked autophagy was triggered by LP formation but not Ca(2+) influx or P28482 - P27361 phosphorylation , 2 canonical Q99572 -evoked signals . Phosphoproteomics , protein expression inference and signaling pathway prediction analysis of Q99572 signaling mediators pointed to P54652 and HSP90 proteins . Indeed , specific HSP90 inhibitors prevented LP formation , LC3-II accumulation , and cell death in myoblasts and myotubes but not in macrophages . Pharmacological blockade or genetic ablation of p2rx7 also proved protective against DB00171 -induced death of muscle cells , as did inhibition of autophagy with 3-MA . The functional significance of the Q99572 LP is one of the great unknowns of purinergic signaling . Our data demonstrate a novel outcome -- autophagy -- and show that molecules entering through the LP can be targeted to phagophores . Moreover , we show that in muscles but not in macrophages , autophagy is needed for the formation of this LP . Given that Q99572 -dependent LP and HSP90 are critically interacting in the DB00171 -evoked autophagic death of dystrophic muscles , treatments targeting this axis could be of therapeutic benefit in this debilitating and incurable form of muscular dystrophy . Surface expression of P48058 AMPA receptor is dependent on an interaction between its C-terminal domain and a 4.1 protein . Dynamic regulation of the number and activity of AMPA receptors is believed to underlie many forms of synaptic plasticity and is presumably mediated by specific protein-protein interactions involving the C-terminal domain of the receptor . Several proteins interacting with the C-terminal tails of the glutamate receptor ( GluR ) -A and P42262 subunits have been identified and implicated in the regulation of endocytosis and exocytosis , clustering , and anchoring of AMPA receptors to the cytoskeleton . In contrast , little is known of the molecular interactions of the P48058 subunit , or of the mechanisms regulating the traffic of P48058 -containing AMPA receptors . We analyzed the subcellular localization of homomeric P48058 receptors carrying C-terminal deletions in transfected human embryonic kidney ( P29320 ) 293 cells and in primary neurons by immunofluorescence microscopy and ELISA . A minimal requirement for a 14-residue cytoplasmic segment for the surface expression of homomeric P48058 receptors was identified . Previously , a similar region in the P42261 subunit was implicated in an interaction with 4.1 family proteins . Coimmunoprecipitation demonstrated that P48058 associated with 4.1 protein(s) in both HEK293 cells and rat brain . Moreover , glutathione S-transferase pull-down experiments showed that the same 14-residue segment is critical for 4.1 binding to P42261 and P48058 . Point mutations within this segment dramatically decreased the surface expression of P48058 in HEK293 cells , with a concomitant loss of the 4.1 interaction . Our findings demonstrate a novel molecular interaction for the P48058 subunit and suggest that the association with the 4.1 family protein(s) plays an essential role in the transport to and stabilization of P48058 -containing AMPA receptors at the cell surface . Examining the genetic and neural components of cognitive flexibility using mice . This commentary summarizes the research presented during the symposium " Examining the genetic and neural components of cognitive flexibility using mice " at the annual meeting of the International Behavioral Neuroscience Society 2011 . Research presented includes examining : 1 ) Corticostriatal networks underlying reversal learning using Q13224 knockout mice , cFos expression , and in vivo electrophysiological recording ; 2 ) Cerebellar contribution to reversal learning using mutants with Purkinje cell loss and in vivo electrochemical recording ; 3 ) Parvalbumin contribution to reversal learning and set-shifting using Q03405 mutants and in vitro recording to examine fast-spiking interneurones ; and 4 ) Alpha 7 nAChR contribution to reversal learning , set-shifting , motivation , and the ' eureka moment ' of rule acquisition . It is proposed that these studies revealed more about the neurobiology underlying these behaviors than could be discovered using pharmacological techniques alone . Together , the research presented stressed the importance of exploring the genetic contribution to neuropsychiatric disease and the important role that the mouse , coupled with robust behavioral measures , can play in understanding neurobiology underlying cognitive flexibility . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Synthesis and structure-activity relationships of novel , substituted 5,6-dihydrodibenzo[a,g]quinolizinium Q99572 antagonists . Iminium quaternary protoberberine alkaloids ( QPA ) have been found to be novel P2X(7) antagonists . To assess their structure-activity relationships , these compounds were modified at their R(1) and R(2) groups and assayed for their ability to inhibit the 2'(3')-O-(4-benzoylbenzoyl)- DB00171 ( BzATP ) -induced uptake of fluorescent ethidium by P29320 -293 cells stably expressing the human P2X(7) receptor , and their ability to inhibit BzATP-induced IL-1beta release by differentiated THP-1 cells . Compounds 15a and 15d , with alkyl groups at the R(1) position , and especially compound 19h , with the 2-NO(2)-4,5-dimethoxy-benzyl group at the R(2) position , had potent inhibitory efficacy as P2X(7) antagonists . Synergistic effects of genetic variation in nicotinic and muscarinic receptors on visual attention but not working memory . It is widely appreciated that neurotransmission systems interact in their effects on human cognition , but those interactions have been little studied . We used genetics to investigate pharmacological evidence of synergisms in nicotinic/muscarinic interactions on cognition . We hypothesized that joint influences of nicotinic and muscarinic systems would be reflected in cognitive effects of normal variation in known SNPs in nicotinic ( P43681 rs1044396 ) and muscarinic ( P08172 rs8191992 ) receptor genes . Exp . 1 used a task of cued visual search . The slope of the cue size/reaction time function showed a trend level effect of the muscarinic P08172 SNP , no effect of the nicotinic P43681 SNP , but a significant interaction between the 2 SNPs . Slopes were steepest in individuals who were both P43681 C/C and P08172 T/T homozygotes . To determine the specificity of this synergism , Exp . 2 assessed working memory for 1-3 locations over 3 s and found no significant effects on either SNP . Interpreting these results in light of Sarter 's [ Briand LA , et al. ( 2007 ) Modulators in concert for cognition : Modulator interactions in the prefrontal cortex. Prog Neurobiol 83:69-91 ] claims of tonic and phasic modes of cholinergic activity , we argue that reorienting attention to the target after invalid cues requires a phasic response , dependent on the nicotinic system , whereas orienting attention to valid cues requires a tonic response , dependent on the muscarinic system . Consistent with that , shifting and scaling after valid cues ( tonic ) were strongest in P43681 C/C homozygotes who were also P08172 T/T homozygotes . This shows synergistic effects within the human cholinergic system . Computational modeling of the direct hydride transfer mechanism for the MAO catalyzed oxidation of phenethylamine and benzylamine : ONIOM ( QM/QM ) calculations . Monoamine oxidases are two isozymic flavoenzymes which are the important targets for drugs used in the treatment of depression , Parkinson and Alzheimer 's diseases . The catalytic reaction taking place between the cofactor DB03147 and amine substrate is still not completely understood . Herein we employed quantum chemical methods on the recently proposed direct hydride transfer mechanism including full active site residues of MAO isoforms in the calculations . Activation free energy barriers of direct hydride transfer mechanism for P21397 and P27338 were calculated by ONIOM ( our own n-layered integrated molecular orbital + molecular mechanics ) method with QM/QM ( quantum mechanics:quantum mechanics ) approach employing several density functional theory functionals , B3LYP , WB97XD , P62158 -B3LYP and M06-2X , for the high layer . The formation of very recently proposed αC-flavin N5 adduct inside the enzyme has been investigated . ONIOM ( M06-2X/6-31+G(d,p):PM6 ) results revealed that such an adduct may form only in P27338 suggesting slightly different hydride transfer mechanisms for P21397 and P27338 . 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 . 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 . 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 . Financial and psychological risk attitudes associated with two single nucleotide polymorphisms in the nicotine receptor ( P43681 ) gene . With recent advances in understanding of the neuroscience of risk taking , attention is now turning to genetic factors that may contribute to individual heterogeneity in risk attitudes . In this paper we test for genetic associations with risk attitude measures derived from both the psychology and economics literature . To develop a long-term prospective study , we first evaluate both types of risk attitudes and find that the economic and psychological measures are poorly correlated , suggesting that different genetic factors may underlie human response to risk faced in different behavioral domains . We then examine polymorphisms in a spectrum of candidate genes that affect neurotransmitter systems influencing dopamine regulation or are thought to be associated with risk attitudes or impulsive disorders . Analysis of the genotyping data identified two single nucleotide polymorphisms ( SNPs ) in the gene encoding the alpha 4 nicotine receptor ( P43681 , rs4603829 and rs4522666 ) that are significantly associated with harm avoidance , a risk attitude measurement drawn from the psychology literature . Novelty seeking , another risk attitude measure from the psychology literature , is associated with several P21964 ( catechol-O-methyl transferase ) SNPs while economic risk attitude measures are associated with several Q05940 ( vesicular monoamine transporter ) SNPs , but the significance of these associations did not withstand statistical adjustment for multiple testing and requires larger cohorts . These exploratory results provide a starting point for understanding the genetic basis of risk attitudes by considering the range of methods available for measuring risk attitudes and by searching beyond the traditional direct focus on dopamine and serotonin receptor and transporter genes . Impact of human D398N single nucleotide polymorphism on intracellular calcium response mediated by α3β4α5 nicotinic acetylcholine receptors . The human P30532 D398N polymorphism ( rs16969968 ) causes an aspartic acid to asparagine change in the nicotinic acetylcholine receptor ( nAChR ) α5 subunit gene . The N398 variant of P30532 is linked to increased risk for nicotine dependence . In this study , we explored the effect of the P30532 D398N polymorphism on the properties of human α3β4* nicotinic acetylcholine receptors in human embryonic kidney ( P29320 ) cells . Addition of either D398 or N398 variant of α5 subunit in the α3β4* receptor did not affect total [ (125)I ] -epibatidine binding or surface expression of the receptor . However , addition of α5(D398) into α3β4* receptor decreased the maximal response to agonist without significantly affecting EC(50) in aequorin intracellular calcium assay . α3β4α5(N398) nAChRs showed further decreased maximal response . The differences in agonist efficacy between the receptor subtypes were found to be dependent upon the concentration of external calcium but independent of external sodium . Moreover , activation of α3β4α5 nAChRs led to significantly greater intracellular calcium release from IP(3) stores relative to α3β4 nAChRs although no effect of the α5 polymorphism was observed . Finally , inclusion of the α5 variant caused a small shift to the left in IC(50) for some of the antagonists tested , depending upon α5 variant but did not affect sensitivity of α3β4* receptors to desensitization in response to incubation with nicotine . In conclusion , addition of either variant of α5 into an α3β4α5 receptor similarly effects receptor pharmacology and function . However , the N398 variant exhibits a reduced response to agonists when extracellular calcium is high and it may lead to distinct downstream cellular signaling . 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 . 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 . Identification of novel genes regulated in the developing human ventral mesencephalon . In the human embryo , from approximately 6 weeks gestational age ( GA ) , dopaminergic ( DA ) neurons can be found in the ventral mesencephalon ( VM ) . More specifically , the post-mitotic neurons are located in the ventral part of the tegmentum ( VT ) , whereas no mature DA neurons are found in the neighboring dorsal part . We used Affymetrix HG-U133 GeneChip technology to compare genome-wide expression profiles of ventral and dorsal tegmentum from 8 weeks GA human embryos , in order to identify genes involved in specification , differentiation , and survival of mesencephalic DA ( mDA ) neurons . Known mDA marker genes including P00352 , Q01959 , Q05940 , TH , P05937 , P43354 , P55317 , P48051 , O75364 , P07949 , and P14416 topped the list of 96 genes from HG-U133A with higher expression in VT , validating the experimental set-up . In addition , 28 probes from HG-U133B were identified whereof most are annotated to UniGene clusters with no gene associated or to genes of unknown function . Of these , the fifteen most regulated transcripts , representing changes down to 56 % could be verified by quantitative real-time PCR ( Q-PCR ) on a developmental series of subdissected human embryonic and fetal brain material , resulting in not only a regional but also a temporal expression profile . This revealed a distinct DA-associated profile for in particular a putative transcription factor ( FLJ45455 ) and the uncharacterized transmembrane proteins Q9ULS5 and Q96EP9 . The data presented here may help to device cell replacement and regenerative therapies for Parkinson 's disease ( PD ) . Complex formation with the Type B gamma-aminobutyric acid receptor affects the expression and signal transduction of the extracellular calcium-sensing receptor . Studies with P29320 -293 cells and neurons . We co-immunoprecipitated the Ca(2+)-sensing receptor ( CaR ) and type B gamma-aminobutyric acid receptor ( GABA-B-R ) from human embryonic kidney ( P29320 ) -293 cells expressing these receptors and from brain lysates where both receptors are present . CaRs extensively co-localized with the two subunits of the GABA-B-R ( Q96GN5 and R2 ) in P29320 -293 cell membranes and intracellular organelles . Coexpressing CaRs and GABA-B-R1s in P29320 -293 cells suppressed the total cellular and cell surface expression of CaRs and inhibited phospholipase C activation in response to high extracellular [ Ca(2+) ] ( [Ca(2+)](e) ) . In contrast , coexpressing CaRs and GABA-B-R2s enhanced CaR expression and signaling responses to raising [Ca(2+)](e) . The latter effects of the O75899 on the CaR were blunted by coexpressing the Q9UBS5 . Coexpressing the CaR with Q9UBS5 or R2 enhanced the total cellular and cell surface expression of the Q9UBS5 or R2 , respectively . Studies with truncated CaRs indicated that the N-terminal extracellular domain of the CaR participated in the interaction of the CaR with the Q9UBS5 and R2 . In cultured mouse hippocampal neurons , CaRs co-localized with the Q9UBS5 and R2 . CaRs and GABA-B-R1s also co-immunoprecipitated from brain lysates . The expression of the CaR was increased in lysates from Q9UBS5 knock-out mouse brains and in cultured hippocampal neurons with their Q9UBS5 genes deleted in vitro . Thus , CaRs and GABA-B-R subunits can form heteromeric complexes in cells , and their interactions affect cell surface expression and signaling of CaR , which may contribute to extracellular Ca(2+)-dependent receptor activation in target tissues . 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 . 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 . 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 . Low ethanol concentration alters P30532 RNA levels during early human development . DB00898 use is common and consumption during pregnancy has been shown to lead to a myriad of physical and neurologic abnormalities commonly referred to as fetal alcohol spectrum disorder . Substance addiction , which includes alcohol , has been shown to involve the major nicotinic acetylcholine receptor subunit P30532 . Using human embryonic stem cells as a model of early human development , we show that low concentrations of ethanol ( 20mM ) can alter the expression of P30532 . Changes in P30532 expression is linked to altered GABA and DB01221 receptor expression , as well as abnormal development of the frontal cortex . These results suggest that alcohol exposure can alter early neurologic development , which may favor addiction and other developmental abnormalities in unborn children . GABA Receptors Genes Polymorphisms and DB00898 Dependence : No Evidence of an Association in an Italian Male Population . OBJECTIVE : The genes encoding for gamma-aminobutyric acid ( GABA ) A and B receptors may be considered as candidates for alcoholism ; genetic alterations at this level may produce structural and functional diversity and thus play a role in the response to alcohol addiction treatment . To investigate these aspects further , we conducted a preliminary genetic association study on a population of Italian male alcohol addicts , focusing on GABA A and B receptors . METHODS : A total of 186 alcohol-dependent subjects ( in the first phase 139 , then 47 more samples ) and 182 controls were genotyped for 25 single nucleotide polymorphisms ( SNPs ) of genes encoding the alpha-1 subunit of GABA A receptor ( P14867 ) and subunits 1 and 2 of GABA B receptor ( Q9UBS5 and O75899 ) . The chi-squared test for allele and genotype distributions and Hardy-Weinberg equilibrium analysis of both subjects and controls were performed . Bonferroni 's correction for multiple comparisons was applied . RESULTS : Preliminary results comparing 139 alcohol-dependent subjects and 182 controls showed differences in genotype distribution in the former for SNP rs29253 , located in the intron region of the Q9UBS5 gene . In order to clarify the meaning of this association , 47 more samples from alcohol-dependent subjects were tested for this SNP only : the previously found association was not confirmed . CONCLUSION : The lack of significant differences between the two groups does not provide evidence that GABRA 1 and Q9UBS5 and 2 genes are candidates for alcoholism in this population . Further studies with larger samples are needed , together with investigation of other components of the GABA pathway . Incorporating age at onset of smoking into genetic models for nicotine dependence : evidence for interaction with multiple genes . DB00184 dependence is moderately heritable , but identified genetic associations explain only modest portions of this heritability . We analyzed 3369 SNPs from 349 candidate genes and investigated whether incorporation of SNP-by-environment interaction into association analyses might bolster gene discovery efforts and prediction of nicotine dependence . Specifically , we incorporated the interaction between allele count and age at onset of regular smoking ( AOS ) into association analyses of nicotine dependence . Subjects were from the Collaborative Genetic Study of DB00184 Dependence and included 797 cases ascertained for Fagerström nicotine dependence and 811 non-nicotine-dependent smokers as controls , all of European descent . Compared with main effect models , SNP x AOS interaction models resulted in higher numbers of nominally significant tests , increased predictive utility at individual SNPs and higher predictive utility in a multi-locus model . Some SNPs previously documented in main effect analyses exhibited improved fits in the joint analysis , including rs16969968 from P30532 and rs2314379 from Q9Y6R4 . P30532 exhibited larger effects in later-onset smokers , in contrast with a previous report that suggested the opposite interaction ( Weiss et al. 2008 ) . However , a number of SNPs that did not emerge in main effect analyses were among the strongest findings in the interaction analyses . These include SNPs located in Q13224 ( P = 1.5 x 10(-5) ) , which encodes a subunit of the N-methyl-D-aspartate receptor channel , a key molecule in mediating age-dependent synaptic plasticity . Incorporation of logically chosen interaction parameters , such as AOS , into genetic models of substance use disorders may increase the degree of explained phenotypic variation and constitutes a promising avenue for gene discovery . P05231 prevents DB01221 -induced neuronal Ca2+ overload via suppression of IP3 receptors . PRIMARY OBJECTIVE : The mechanism underlying interleukin-6 ( P05231 ) prevention of N-methyl-D-aspartate ( DB01221 ) -induced neuronal Ca(2+) overload was explored at the profile of Ca(2+) channel receptors , including DB01221 , inositol 1,4,5-trisphosphate and ryanodine receptors ( NMDAR , IP3R and RyR , respectively ) . METHODS : Cerebellar granule neurons from 8-day-old rats were exposed to P05231 ( 40 or 120 ng ml(-1) ) for 8 days and stimulated with DB01221 ( 100 μM ) for 15 or 30 minutes . RESULTS : DB01221 evoked an acute and sustained enhancement of intracellular Ca(2+) fluorescence intensity in the entire 15-minute DB01221 application period . P05231 prevented the acute and sustained intracellular Ca(2+) elevation triggered by DB01221 in a concentration-dependent manner . MK-801 , an NMDAR antagonist , completely suppressed DB01221 -evoked neuronal Ca(2+) overload in the absence or presence of P05231 . IP3R antagonist 2- Q9H4A4 lessened DB01221 -evoked acute and sustained cytosolic Ca(2+) overload and P05231 further reduced the acute 2- Q9H4A4 -dependent Ca(2+) component . Dissimilarly , after RyR antagonist DAN treatment , DB01221 still induced an acute and sustained elevation of intracellular Ca(2+) levels , and the elevated Ca(2+) was significantly suppressed by P05231 . Moreover , P05231 down-regulated Q05586 and IP3R1 but did not alter Q92736 expression . CONCLUSION : The present results suggest that P05231 suppresses DB01221 -induced neuronal Ca(2+) overload by inhibiting NMDAR and IP3R activities . 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 . 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 . PI 3-kinase and PKCζ mediate insulin-induced potentiation of DB01221 receptor currents in Xenopus oocytes . P01308 modulates N-methyl-d-aspartate ( DB01221 ) receptors in the CNS and potentiates recombinant DB01221 receptor currents in Xenopus oocytes . We have previously found that insulin 's potentiation of DB01221 receptor currents in oocytes occurs in a subunit specific manner and via phosphorylation of specific C-terminal sites by protein tyrosine kinases ( PTKs ) and C-type protein kinases ( PKCs ) . P01308 -mediated current potentiation of receptors containing the Q12879 subunit occurs solely through the activation of PKCs . Activation of phosphoinositide 3-kinase ( PI 3-kinase ) is known to trigger many insulin-stimulated signaling pathways , and we show here that it lies at a critical step in the insulin-mediated potentiation of DB01221 receptor currents . Incubation with the PI 3-kinase inhibitor wortmannin eliminates insulin potentiation of DB01221 receptor currents in the oocytes . Atypical isoforms of PKC are known to be activated downstream in the insulin signaling pathway via activation of PI 3-kinase . We demonstrate that the atypical isoform PKC zeta ( PKCζ ) has a role in insulin-stimulated current potentiation of Q12879 -containing DB01221 receptors using an isoform-specific pseudosubstrate inhibitor of PKCζ . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . 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 . 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 . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Human plasmacytoid dendritic cells express an atrial natriuretic peptide receptor , guanylyl cyclase-A . Atrial natriuretic peptide is a cardiovascular hormone secreted mainly by the cardiac atria and regulates the volume-pressure homeostasis . The action of P01160 is mediated by P16066 . We previously reported that human monocyte-derived dendritic cells express P16066 and respond to P01160 with polarization toward a Th2-inducing phenotype . In the present study , we explored the possibility that pDC are subjected to immunoregulation via the P01160 / P16066 system . We examined P16066 expression on blood pDC and found that P16066 was not expressed on fresh pDC but was induced after stimulation with CpG-oligodeoxynucleotide AAC-30 , P08700 , or interleukin-3 plus P29965 . Activated pDC responded to P01160 with an increase in cGMP production , indicating that P16066 expressed on pDC was functional . We investigated whether tonsillar pDC express P16066 by immunohistochemistry and immunofluorescence staining . We found that P16066 (+) HLA-DR(+) cells were present in the T-cell areas and the perivascular areas . Flow cytometric analysis with tonsillar cells confirmed that lineage(-) CD123(high) pDC express P16066 . These results indicate that the P01160 / P16066 system is involved in immune regulation through pDC in secondary lymphoid organs . Large candidate gene association study reveals genetic risk factors and therapeutic targets for fibromyalgia . OBJECTIVE : Fibromyalgia ( FM ) represents a complex disorder that is characterized by widespread pain and tenderness and is frequently accompanied by additional somatic and cognitive/affective symptoms . Genetic risk factors are known to contribute to the etiology of the syndrome . The aim of this study was to examine > 350 genes for association with FM , using a large-scale candidate gene approach . METHODS : The study group comprised 496 patients with FM ( cases ) and 348 individuals with no chronic pain ( controls ) . Genotyping was performed using a dedicated gene array chip , the Pain Research Panel , which assays variants characterizing > 350 genes known to be involved in the biologic pathways relevant to nociception , inflammation , and mood . Association testing was performed using logistic regression . RESULTS : Significant differences in allele frequencies between cases and controls were observed for 3 genes : P28472 ( rs4906902 ; P = 3.65 × 10(-6) ) , Q96RJ0 ( rs8192619 ; P = 1.11 × 10(-5) ) , and P32455 ( rs7911 ; P = 1.06 × 10(-4) ) . These 3 genes and 7 other genes with suggestive evidence for association were examined in a second , independent cohort of patients with FM and control subjects who were genotyped using the Perlegen 600K platform . Evidence of association in the replication cohort was observed for Q96RJ0 , P49798 , P21554 , and P48058 . CONCLUSION : Variation in these 4 replicated genes may serve as a basis for development of new diagnostic approaches , and the products of these genes may contribute to the pathophysiology of FM and represent potential targets for therapeutic action . 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 . 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 . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Association study of 45 candidate genes in nicotine dependence in Han Chinese . Numerous genetic linkages , association studies have been performed in different ethnic groups and revealed many susceptibility loci and genes for nicotine dependence . However , limited similar researches were performed in Han Chinese . This study was designed to investigate the association of candidate genes with nicotine dependence in Han Chinese . We genotyped 384 SNPs within 45 candidate genes with nicotine dependence in a Han Chinese population consisting 223 high nicotine dependent subjects and 257 low nicotine dependent subjects by employing GoldenGate genotyping assay ( Illumina ) . Following association analysis was performed using PLINK software . Individual SNP-based association analysis revealed that nine SNPs located in P35462 ( rs2630351 ) , P21918 ( rs1967550 ) , Q9Y6R4 ( rs2314378 ) , DDC ( rs11575461 ) , Q05901 ( rs4954 ) , O75899 ( rs2779562 ) , P14416 ( rs11214613 and rs6589377 ) and P43681 ( rs2236196 ) were significantly associated with FTND after correction for multiple testing with the p values from 2.59×10(-7) to 9.99×10(-5) . Haplotype-based association analysis revealed haplotype G-A-A formed by rs2630351 , rs167771 and rs324032 and haplotype G-G-G-A formed by rs3773678 , rs2630349 , rs2630351 and rs167771 in P35462 ; haplotype of G-A formed by rs2779562 and rs2808566 in O75899 and haplotype of T-T-A-G-A formed by rs6832644 , rs4057797 , rs9764 , rs4552421 and rs10033119 in P25929 are associated with FTND ( p=3.61×10(-7)-8.78×10(-6) ) . Our results provided confirmation of the previous findings that P14416 , P35462 , DDC , Q05901 , O75899 and P43681 are associated with nicotine dependence . Furthermore , we for the first time report a significant association between nicotine dependence and P21918 , Q9Y6R4 and P25929 . These findings need independent replication in the future studies . 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 . 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 . 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 . 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 . 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 . 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 . 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 . DB01184 treatment for gastroparesis : demographic and pharmacogenetic characterization of clinical efficacy and side-effects . BACKGROUND : DB01184 is a useful alternative to metoclopramide for treatment of gastroparesis due to better tolerability . Effectiveness and side-effects from domperidone may be influenced by patient-related factors including polymorphisms in genes encoding drug-metabolizing enzymes , drug transporters , and domperidone targets . AIMS : The aim of this study was to determine if demographic and pharmacogenetic parameters of patients receiving domperidone are associated with response to treatment or side-effects . METHODS : Patients treated with domperidone for gastroparesis provided saliva samples from which DNA was extracted . Fourteen single-nucleotide polymorphisms ( SNPs ) in seven candidate genes ( P08183 , P10635 , P14416 , P15382 , Q9Y6J6 , Q12809 , P51787 ) were used for genotyping . SNP microarrays were used to assess single-nucleotide polymorphisms in the ADRA1A , P35368 , and P25100 loci . RESULTS : Forty-eight patients treated with domperidone participated in the study . DNA was successfully obtained from each patient . Age was associated with effectiveness of domperidone ( p=0.0088 ) . Genetic polymorphism in Q12809 was associated with effectiveness of domperidone ( p=0.041 ) . The efficacious dose was associated with polymorphism in P08183 gene ( p=0.0277 ) . The side-effects of domperidone were significantly associated with the SNPs in the promoter region of P25100 gene . CONCLUSIONS : Genetic characteristics associated with response to domperidone therapy included polymorphisms in the drug transporter gene P08183 , the potassium channel Q12809 gene , and α1D -- adrenoceptor P25100 gene . Age was associated with a beneficial response to domperidone . If verified in a larger population , this information might be used to help determine which patients with gastroparesis might respond to domperidone and avoid treatment in those who might develop side-effects . 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 . 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 . 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 . 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 . DB00989 improves hippocampal neurogenesis and depression-like behaviors via P08908 receptor stimulation in olfactory bulbectomized mice . DB00989 is a non-competitive inhibitor of both acetylcholinesterase ( P22303 ) and butylcholinesterase ( BuChE ) used to treat mild to moderate dementia in Alzheimer 's disease ( AD ) patients . Although rivastigmine reportedly ameliorates cognitive dysfunction in these patients , its ability to improve Behavioral and Psychological Symptoms of Dementia ( BPSD ) remains unclear . To determine whether rivastigmine treatment antagonizes depression-like behaviors , we chronically administered rivastigmine ( 0.1-1.0mg/kg ) to olfactory bulbectomized ( OBX ) mice once a day for 2weeks , starting 2weeks after bulbectomy . Chronic treatment at 0.3 or 1.0mg/kg dose dependently and significantly improved depression-like behaviors , as assessed by tail suspension ( Q16762 ) , forced swim ( P19883 ) , locomotion and novelty-suppressed feeding ( NSFT ) tests . Importantly , co-administration with WAY-100635 ( 1.0mg/kg ) , a P08908 receptor antagonist , but not ketanserin ( 1.0mg/kg , ) , a 5- Q13049 receptor antagonist , completely blocked rivastigmine-induced anti-depressive effects , suggesting that P08908 receptor stimulation mediates this activity . Consistent with this observation , rivastigmine treatment significantly rescued impaired neurogenesis observed in OBX mice in a P08908 receptor-dependent manner . Furthermore , enhanced protein kinase B ( Akt ) and extracellular signal-regulated kinase ( P29323 ) phosphorylation seen following rivastigmine treatment was closely associated with improved neurogenesis . These effects were blocked by WAY-100635 but not ketanserin treatment . Finally , we confirmed that P08908 but not 5- Q13049 receptor stimulation by specific agonists mimicked rivastigmine-induced anti-depression activity and promoted hippocampal neurogenesis . We conclude that , in addition to enhancing the cholinergic system , rivastigmine treatment restores normal function of the hippocampal serotonergic system , an activity that likely ameliorates depressive behaviors in AD patients . 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 . [ 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 .

MH_train_1139

MH_train_1139

MH_train_1139

interacts_with DB08895?

multiple_choice

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 . 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 . Polymorphisms in JAK/ P35610 signaling pathway genes and risk of non-Hodgkin lymphoma . Impaired function of Janus kinase/signal transducer and activator of transcription ( JAK/ P35610 ) signaling pathway genes leads to immunodeficiency and various hematopoietic disorders . We evaluated the association between genetic polymorphisms ( SNPs ) in 12 JAK/ P35610 pathway genes ( P52333 , P42224 , P52630 , P40763 , Q14765 , STAT5a , STAT5b , P42226 , SCOS1 , SCOS2 , SCOS3 , and SCOS4 ) and Q9NZ71 risk in a population-based case-control study of Connecticut women . We identified three SNPs in P40763 ( rs12949918 and rs6503695 ) and Q14765 ( rs932169 ) associated with Q9NZ71 risk after adjustment for multiple comparison . Our results suggest that genetic variation in JAK/ P35610 pathway genes may play a role in lymphomagenesis and warrants further investigation . Two-sided roles of Q8TAD2 : induction of Th1 differentiation on naive P01730 + T cells versus suppression of proinflammatory cytokine production including IL-23-induced Q16552 on activated P01730 + T cells partially through P40763 -dependent mechanism . Recent lines of evidence have demonstrated that Q8TAD2 , a newly identified IL-12-related cytokine , has two apparently conflicting roles in immune responses : one as an initiator of Th1 responses and the other as an attenuator of inflammatory cytokine production . Although the Q8TAD2 -mediated Th1 initiation mechanism has been elucidated , little is known about the molecular basis for the suppression of cytokine production . In the present study , we demonstrated that Q8TAD2 suppressed the production of various proinflammatory cytokines by fully activated P01730 + T cells while it had no effect on the cytokine production by P01730 + T cells at early phases of activation . Q8TAD2 also suppressed Q16552 production by activated P01730 + T cells , thereby counteracting IL-23 , another IL-12-related cytokine with proinflammatory effects . In fully activated P01730 + T cells , P40763 was preferentially activated by Q8TAD2 stimulation , whereas both P42224 and 3 were activated by Q8TAD2 in early activated P01730 + T cells . Lack of P40763 in fully activated cells impaired the suppressive effects of Q8TAD2 . These data indicated that the preferential activation of P40763 in fully activated P01730 + T cells plays an important role in the cytokine suppression by Q8TAD2 /WSX-1 . 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 . Contact sensitization to oxazolone : involvement of both interferon-gamma and interleukin-4 in oxazolone-specific Ig and T-cell responses . The synthesis and role of several lymphokines were examined during contact sensitization to oxazolone ( OX ) . Application of OX to the skin of mice increased the delayed-type hypersensitivity ( DTH ) response to challenge , serum titres of OX-specific IgG1 and IgG2a , and draining lymph node cell ( LNC ) numbers . At day 3 , LN contained detectable interleukin-4 ( P05112 ) , interferon-gamma ( P01579 ) and granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) but not P60568 or P08700 mRNAs ; P08700 and higher levels of P05112 , P01579 and GM- P04141 mRNAs were measured after 24 hr culture with anti-CD3 antibody in OX-primed but not unprimed LNC . As a result of sensitization , LNC secreted P08700 constitutively and produced elevated levels of P60568 , P08700 , P05112 and P01579 in response to anti-CD3 antibody ; a similar but weaker lymphokine response was recalled by OX-protein conjugate . P01730 + cells were the major source of the anti-CD3-induced lymphokines except P01579 , which was derived mainly from CD8+ cells . Since both P05112 and P01579 were synthesized by OX-primed LNC in vivo and in vitro , their role was investigated by administering anti-lymphokine antibodies at the time of sensitization . Anti- P05112 treatment reduced OX-specific serum IgG1 titres without affecting IgG2a titres , whereas anti- P01579 treatment reduced IgG2a but not IgG1 titres . Although neither antibody altered DTH responsiveness , anti- P01579 treatment markedly increased P05112 production by P01730 + LNC and reduced P01579 production in vitro , particularly by P01730 + cells . We conclude that endogenous P05112 and P01579 reciprocally influence the isotype of the Ig response to OX and that P01579 also affects the relative levels of P05112 and P01579 synthesis by P01730 + LNC . 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 . 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 . 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 . 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 . Effect of targeting janus kinase 3 on the development of intestinal tumors in the adenomatous polyposis coli(min) mouse model of familial adenomatous polyposis . Familial adenomatous polyposis ( FAP ) is associated with germ-line mutations in the tumor suppressor gene , adenomatous polyposis coli ( P25054 ) located on chromosome 5q21 . Multiple intestinal neoplasia ( Min ) in mice resembles FAP in humans , resulting from a single point mutation in the murine homolog of the P25054 gene . The effects of the rationally-designed P52333 ( P52333 ) inhibitor JANEX-1 ( 4-(4'-hydroxyphenyl) amino-6,7-dimethoxy-quinazoline , WHI-P131 , CAS 202475-60-3 ) on the development of intestinal tumors in the P25054 ( min/+ ) mouse model of FAP were examined . The Min mice were fed with rodent chow or chow supplemented with JANEX-1 once a week starting at 1.5 months of age . The cumulative proportions of mice remaining alive at 7 months were 13 +/- 6 % for control mice versus 72 +/-12 % for JANEX-1 treated mice ( P < 0.0002 ) . In contrast , Compound DDE24 , a synthetically activated genistein , an inhibitor of Epidermal Growth Factor receptor ( P01133 -R ) , cellular homologue of oncogene product from Raus Avian sarcoma virus ( P12931 ) and Syk tyrosine kinases , which Thus , selective targeting of P52333 was highly effective in preventing development of intestinal tumors in Min mice resulting in markedly improved survival outcomes . P52333 inhibitors may therefore be useful in the prevention of colorectal cancer in individuals with FAP . 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 . 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 . 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 . P52333 activity is necessary for phosphorylation of cytosolic phospholipase A2 and prostaglandin E2 synthesis by macrophages infected with Francisella tularensis live vaccine strain . Francisella tularensis , the causative agent of tularemia , modulates the host immune response to gain a survival advantage within the host . One mechanism of immune evasion is the ability of F. tularensis to induce the synthesis of the small lipid mediator prostaglandin E2 ( DB00917 ) , which alters the host T cell response making the host more susceptible to Francisella growth . DB00917 is synthesized by a tightly regulated biosynthetic pathway following stimulation . The synthesis of DB00917 begins with the liberation of arachidonic acid ( AA ) from membrane phospholipids by cytosolic phospholipase A2 ( P47712 ) . AA is subsequently converted to the unstable intermediate PGH2 by cyclooxygenase-2 ( P35354 ) , and PGH2 undergoes an isomerization reaction to generate DB00917 . Our objective was to identify F. tularensis-activated host signaling pathways that regulate the activity of the enzymes in the DB00917 -biosynthetic pathway . In this study , we show that P47712 , p38 mitogen-activated protein kinase ( MAPK ) , and P52333 ( P52333 ) signaling are necessary for F. tularensis-induced DB00917 production . Inhibition of P52333 activity reduced the phosphorylation of P47712 and P35354 protein levels . In addition , P52333 regulates P47712 phosphorylation independent of transcription . Moreover , p38 MAPK activity is required for F. tularensis-induced P35354 protein synthesis , but not for the phosphorylation of P47712 . This research highlights a unique signaling axis in which P52333 and p38 MAPK regulate the activity of multiple enzymes of the DB00917 -biosynthetic pathway in macrophages infected with F. tularensis . 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 . 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 . Differences in signaling through the B-cell leukemia oncoprotein Q9HC73 in response to Q969D9 and through mutant O60674 . Approximately 10 % of B-cell acute lymphoblastic leukemias ( B-ALLs ) overexpress the cytokine receptor subunit Q9HC73 , which may confer a poor prognosis . Q9HC73 binds its ligand thymic stromal lymphopoietin ( Q969D9 ) as a heterodimer with P16871 . Subsets of Q9HC73 -overexpressing B-ALLs also have a gain-of-function Q9HC73 F232C mutation or activating mutations in O60674 . Whether these mutant alleles confer differences in signaling has not been addressed . Through a domain mutation analysis , we demonstrate a distinct dependence on the Q9HC73 intracellular tyrosine Y368 in signaling by Q9HC73 F232C , but not signaling induced by Q969D9 or through Q9HC73 /mutant O60674 . In contrast , Q9HC73 signaling in each context is strictly dependent on both the Q9HC73 box1 domain and the intracellular tryptophan W286 . Using a global quantitative analysis of tyrosine phosphorylation induced by Q969D9 , we previously identified Q969D9 -induced phosphorylation of multiple kinases implicated in B-cell receptor signaling , including Lyn , Btk , Hck , Syk , P45983 , P45984 , and P53779 . We now demonstrate that cells dependent on Q9HC73 /mutant O60674 have reduced phosphorylation at these targets , suggesting that the kinases promote Q969D9 -mediated proliferation but serve as negative regulators of Q9HC73 /mutant O60674 signaling . Thus , targetable nodes downstream of Q9HC73 differ based on the presence or absence of additional mutations in Q9HC73 signaling components . 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 . 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 . 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 . The induction and activation of P42224 by all-trans-retinoic acid are mediated by RAR beta signaling pathways in breast cancer cells . Retinoic acid receptor-beta ( RAR beta ) and signal transducer and activator of transcription 1 ( P42224 ) are important mediators of the antiproliferative and apoptotic actions of retinoids and cytokines/growth factors , respectively . Expression of both RAR beta and P42224 is lost in most breast cancer cell lines but it can be induced by retinoids in estrogen receptor-positive cells . We investigated a possible functional connection between these two mediators and present evidence supporting RAR beta as a tumor suppressor . First , by using different receptor-selective retinoids , we demonstrated that RAR beta induction in MCF-7 cells by all-trans-retinoic acid ( atRA ) was associated with the activation of P42224 gene transcription . The direct involvement of RAR beta in atRA-induced P42224 gene activation was further demonstrated by showing that transfection with an anti-sense RAR beta construct blocked atRA-induced P42224 expression in MCF-7 cells whereas introduction of a sense-RAR beta construct resulted in P42224 induction by atRA in MDA-MB 231 cells . In addition , we showed that P42224 was phosphorylated/activated under atRA treatment of MCF-7 cells ; this process required the involvement of RAR beta and protein synthesis . P42224 phosphorylation/activation was accompanied by increased tyrosine kinase activity that was not due to the activation of P23458 , O60674 or Tyk 2 , suggesting the possible involvement of an unidentified tyrosine kinase . P13232 activates the phosphatidylinositol 3-kinase/AKT pathway in normal human thymocytes but not normal human B cell precursors . P13232 signaling culminates in different biological outcomes in distinct lymphoid populations , but knowledge of the biochemical signaling pathways in normal lymphoid populations is incomplete . We analyzed CD127/IL-7Ralpha expression and function in normal ( nontransformed ) human thymocytes , and human P15391 (+) B-lineage cells purified from xenogeneic cord blood stem cell/MS-5 murine stromal cell cultures , to further clarify the role of P13232 in human B cell development . P13232 stimulation of P28906 (+) immature thymocytes led to phosphorylation ( p- ) of P42229 , P27361 /2 , AKT , and glycogen synthase kinase-3 beta , and increased AKT enzymatic activity . In contrast , P13232 stimulation of P28906 (-) thymocytes ( that included P01730 (+)/CD8(+) double-positive , and P01730 (+) and CD8(+) single-positive cells ) only induced p- P42229 . P13232 stimulation of P15391 (+) cells led to robust induction of p- P42229 , but minimal induction of p- P27361 /2 and p-glycogen synthase kinase-3 beta . However , P15391 (+) cells expressed endogenous p- P27361 /2 , and when rested for several hours following removal from MS-5 underwent de-phosphorylation of P27361 /2 . P13232 stimulation of rested P15391 (+) cells resulted in robust induction of p- P27361 /2 , but no induction of AKT enzymatic activity . The use of a specific P52333 antagonist demonstrated that all P13232 signaling pathways in P28906 (+) thymocytes and P15391 (+) B-lineage cells were P52333 -dependent . We conclude that human P28906 (+) thymocytes and P15391 (+) B-lineage cells exhibit similarities in activation of P42229 and P27361 /2 , but differences in activation of the PI3K/AKT pathway . The different induction of PI3K/AKT may at least partially explain the different requirements for P13232 during human T and B cell development . 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 . Helicobacter pylori-induced P40763 activation and signalling network in gastric cancer . BACKGROUND : Helicobacter pylori ( H. pylori ) is the most important gastric carcinogen . However , the mechanisms of H. pylori induced gastric carcinogenesis through P40763 activation are largely unknown . We evaluated the effects of H. pylori infection on P40763 activation and dissected the signalling network of P40763 in H. pylori- infected gastric carcinogenesis . METHODS : The expression of phospho- P40763 ( pSTAT3 ) was evaluated by immunohistochemistry and western blot . Gene expression array and chromatin immunoprecipitation were used to dissect the P40763 signalling network on H. pylori co-cultured AGS . RESULTS : pSTAT3 was significantly higher in H. pylori -positive gastritis than in H. pylori -negative gastritis ( P = 0.003 ) . In addition , 98 % of H. pylori positive intestinal metaplasia specimens showed P40763 activation , whereas pSTAT3 was significantly decreased in all 43 specimens one year after H. pylori eradication ( P < 0.001 ) . Moreover , pSTAT3 was only detected in the H. pylori -infected gastric tissues of mice but not in control mice . We further identified 6 candidates ( Q9BZC1 , P11362 , O60902 , P52333 , P45983 , and Q86YL7 ) were directly up-regulated by H. pylori induced P40763 activation . CONCLUSION : H. pylori infection triggers the activation of P40763 and de-regulates multitude of tumorigenic genes which may contribute to the initiation and progression of gastric cancer . 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 . 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 . 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 . P05112 up-regulates histamine H1 receptors by activation of H1 receptor gene transcription . DB11320 plays an important role in allergy mainly through histamine H1 receptor ( P35367 ) . Recent studies showed that the P35367 level is elevated in allergic conditions , suggesting that this will make the allergic symptoms worse by intensifying P35367 -mediated processes . Some cytokines are also involved in allergy , and interleukin-4 ( P05112 ) has been implicated as an important mediator of allergic inflammation . It is noteworthy that the level of P05112 is elevated under allergic states . We tested whether P05112 has a role in up-regulating P35367 level by using the cultured human HeLa cell as a model system that expresses both P05112 receptor and P35367 . P05112 stimulation increased P35367 protein levels and P35367 mRNA levels . P05112 also increased P35367 promoter activity , but had no effect on P35367 mRNA stability , indicating that up-regulation of P35367 was due to an increase in P35367 mRNA synthesis . P05112 activated P42226 ( signal transducer and activator of transcription 6 ) in HeLa cells , and up-regulation of P35367 mRNA and activation of P42226 by P05112 were inhibited by a specific P52333 ( Janus-activated kinase 3 ) inhibitor . Stimulation with histamine also up-regulated P35367 mRNA , and co-stimulation with histamine and P05112 elevated P35367 mRNA level significantly higher than the stimulation with histamine or P05112 alone did . These results indicated that P05112 up-regulated P35367 mRNA level through increased transcription of P35367 gene via P52333 - P42226 pathway . The effects of histamine and P05112 were additive , suggesting that these allergic mediators will work together to up-regulate P35367 level , and thus make the allergic symptom worse by intensifying P35367 -mediated allergic processes . 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 . 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 . 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 . [ 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 . GIP-dependent expression of hypothalamic genes . GIP ( glucose dependent insulinotrophic polypeptide ) , originally identified as an incretin peptide synthesized in the gut , has recently been identified , along with its receptors ( P48546 ) , in the brain . Our objective was to investigate the role of GIP in hypothalamic gene expression of biomarkers linked to regulating energy balance and feeding behavior related neurocircuitry . Rats with lateral cerebroventricular cannulas were administered 10 μg GIP or 10 microl artificial cerebrospinal fluid ( aCSF ) daily for 4 days , after which whole hypothalami were collected . Real time Taqman™ RT-PCR was used to quantitatively compare the mRNA expression levels of a set of genes in the hypothalamus . Administration of GIP resulted in up-regulation of hypothalamic mRNA levels of AVP ( 46.9±4.5 % ) , Q16568 ( 25.9±2.7 % ) , P16220 ( 38.5±4.5 % ) , O14764 ( 67.1±11 % ) , O60674 ( 22.1±3.6 % ) , P28482 ( 33.8±7.8 % ) , P01303 ( 25.3±5.3 % ) , P01178 ( 49.1±5.1 % ) , P40763 ( 21.6±3.8 % ) , and TH ( 33.9±8.5 % ) . In a second experiment the same set of genes was evaluated in P48546 (-/-) and P48546 ( +/ ? ) mice to determine the effect of lack of GIP stimulation on gene expression . In P48546 (-/-) mice expressions of the following genes were down-regulated : AVP ( 27.1±7.5 % ) , Q16568 ( 28.3±3.7 % ) , P01178 ( 25.2±5.8 % ) , O14684 ( 23.9±4.5 % ) , and P40763 ( 8.8±2.3 % ) . These results suggest that AVP , Q16568 , P01178 and P40763 may be involved in energy balance-related hypothalamic circuits affected by GIP . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . 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 . 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 . 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 . Association of cigarette smoking and CRP levels with DNA methylation in α-1 antitrypsin deficiency . Alpha-1 antitrypsin ( P01009 ) deficiency and tobacco smoking are confirmed risk factors for Chronic Obstructive Pulmonary Disease . We hypothesized that variable DNA methylation would be associated with smoking and inflammation , as reflected by the level of C-Reactive Protein ( CRP ) in P01009 -deficient subjects . Methylation levels of 1,411 autosomal CpG sites from the Illumina GoldenGate Methylation Cancer Panel I were analyzed in 316 subjects . Associations of five smoking behaviors and CRP levels with individual CpG sites and average methylation levels were assessed using non-parametric testing , linear regression and linear mixed effect models , with and without adjustment for age and gender . Univariate linear regression analysis revealed that methylation levels of 16 CpG sites significantly associated with ever-smoking status . A CpG site in the Q15582 gene was the only site associated with ever-smoking after adjustment for age and gender . No highly significant associations existed between age at smoking initiation , pack-years smoked , duration of smoking , and time since quitting smoking as predictors of individual CpG site methylation levels . However , ever-smoking and younger age at smoking initiation associated with lower methylation level averaged across all sites . DNA methylation at CpG sites in the Q13761 , P52333 and P04264 genes associated with CRP levels . The most significantly associated CpG sites with gender and age mapped to the P55212 and O00144 genes , respectively . In summary , this study identified multiple potential candidate CpG sites associated with ever-smoking and CRP level in P01009 -deficient subjects . Phenotypic variability in Mendelian diseases may be due to epigenetic factors . 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 . 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 . Dysfunctional Immune-Mediated Inflammation in Rheumatoid Arthritis Dictates that Development of Anti-Rheumatic Disease Drugs Target Multiple Intracellular Signaling Pathways . A skewed repertoire of pro-inflammatory cytokines produced by the Th1 subset , one of the hallmarks of rheumatoid arthritis ( RA ) , is characterized by an overabundance of pro-inflammatory cytokines . P01375 -α , interleukin- 1 ( IL-1 ) , P05231 , P13232 , P10145 , Q9HBE4 , IL-12/IL-23 , P40933 , Q16552 , Q14116 , P24001 , and interferon-γ are primarily responsible for immune-mediated inflammation of RA by activating Janus kinases ( JAK ) -1 , -2 , -3 , p38 kinase , C-Jun-Nterminal kinase , extracellular signal-regulated kinase 1/2 and the phosphatidylinosotide-3-kinase/Akt/mTor pathways . Activation of these signaling pathways results in up-regulation of pro-inflammatory cytokines , cyclooxygenase-2 , matrix metalloproteinases , pro-angiogenesis proteins and anti-apoptosis proteins , the latter resulting in abnormal survival of activated T- and B-cells . Further , Q16552 also regulates the differentiation of P01730 + T-helper cells by inducing a Th17 T-cell subset , and a subpopulation of T-regulatory ( Treg ) cells . Although Treg cells are sufficiently abundant in RA synovial fluid , they fail to induce immune tolerance suggesting a functional deficiency likely coupled to putative protein kinase signaling abnormalities . The results of in vitro and studies in animal models of arthritis have indicated that inhibiting individual signaling pathways can blunt the synthesis of several of the pro-inflammatory biomarkers characteristic of human RA pathology . However , RA clinical trials indicated that small molecule inhibitors of P23458 , -2- , 3 and/or p38 kinase while exhibiting acceptable safety and tolerability profiles have only marginal and transient clinical effectiveness . These results suggested that future RA clinical studies using these or other kinase inhibitors will have to consider strategies designed to simultaneously inhibit multiple kinase pathways . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . The kinase inhibitory region of O15524 is sufficient to inhibit T-helper 17 and other immune functions in experimental allergic encephalomyelitis . Suppressors of cytokine signaling ( Q9NSE2 ) negatively regulate the immune response , primarily by interfering with the JAK/ P35610 pathway . We have developed a small peptide corresponding to the kinase inhibitory region ( P55040 ) sequence of O15524 , O15524 - P55040 , which inhibits kinase activity by binding to the activation loop of tyrosine kinases such as O60674 and P29597 . Treatment of SJL/J mice with O15524 - P55040 beginning 12 days post-immunization with myelin basic protein ( MBP ) resulted in minimal symptoms of EAE , while most control treated mice developed paraplegia . O15524 - P55040 treatment suppressed interleukin-17A ( Q16552 ) production by MBP-specific lymphocytes , as well as MBP-induced lymphocyte proliferation . When treated with IL-23 , a key cytokine in the terminal differentiation of Q16552 -producing cells , MBP-sensitized cells produced Q16552 and IFNγ ; O15524 - P55040 was able to inhibit the production of these cytokines . O15524 - P55040 also blocked IL-23 and Q16552 activation of P40763 . There is a deficiency of O15524 and O14543 mRNA expression in P01730 (+) T cells that infiltrate the CNS , reflecting a deficiency in regulation . Consistent with therapeutic efficacy , O15524 - P55040 reversed the cellular infiltration of the CNS that is associated with EAE . We have shown here that a O15524 like effect can be obtained with a small functional region of the O15524 protein that is easily produced . The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 .

MH_train_1140

MH_train_1140

MH_train_1140

interacts_with DB00054?

multiple_choice

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 . 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 . DB00054 : cost-effective survival advantage in clinical trials and clinical practice . Adjunctive blockade of the platelet glycoprotein ( GP ) IIb/IIIa receptor during either percutaneous coronary intervention ( P05154 ) or for patients who present with non-ST segment elevation acute coronary syndromes has demonstrated efficacy in reducing platelet-mediated adverse cardiovascular ischemic events . The three currently available agents ( abciximab , eptifibatide , tirofiban ) differ markedly in pharmacodynamic and pharmacokinetic profiles , receptor affinity , and cost . Although pharmacoeconomic substudies are available from placebo-controlled randomized trials of platelet P08514 /IIIa blockade during P05154 , " real-world " cost-effectiveness data from high-volume practice are lacking . Therefore , in-hospital and late ( 6-month ) clinical outcomes and cumulative cost/charge data were analyzed on 1472 consecutive P05154 procedures ( 70 % received abciximab ) performed by high-volume operators at a single institution.(1) Data were adjusted for lack of randomized treatment allocation with the use of a propensity scoring technique . Adjunctive abciximab therapy for P05154 was associated with a significant ( 3.4 % ) reduction in mortality to 6 months . Based on the economic cost-effectiveness concept of cost per life year gained relative to standard therapy,(2,3) abciximab provided a cost-effective survival advantage in high-volume interventional practice that compares very favorably with currently accepted standards . Clinical and procedural demographics associated with increased cost-effectiveness include multivessel coronary intervention , stent deployment , recent ( < 1 week ) myocardial infarction ( MI ) , and impaired left-ventricular ( LV ) function . 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 . P38398 , Q9BXW9 and Chk1 are potential molecular targets for the modulation of a radiation-induced DNA damage response in bystander cells . Radiotherapy is an important treatment option for many human cancers . Current research is investigating the use of molecular targeted drugs in order to improve responses to radiotherapy in various cancers . The cellular response to irradiation is driven by both direct DNA damage in the targeted cell and intercellular signalling leading to a broad range of bystander effects . This study aims to elucidate radiation-induced DNA damage response signalling in bystander cells and to identify potential molecular targets to modulate the radiation induced bystander response in a therapeutic setting . Stalled replication forks in T98G bystander cells were visualised via bromodeoxyuridine ( BrdU ) nuclear foci detection at sites of single stranded DNA . γ P16104 co-localised with these BrdU foci . P38398 and Q9BXW9 foci formed in T98G bystander cells . Using ATR mutant F02-98 hTERT and Q13315 deficient GM05849 fibroblasts it could be shown that ATR but not Q13315 was required for the recruitment of Q9BXW9 to sites of replication associated DNA damage in bystander cells whereas P38398 bystander foci were Q13315 -dependent . Phospho-Chk1 foci formation was observed in T98G bystander cells . Clonogenic survival assays showed moderate radiosensitisation of directly irradiated cells by the Chk1 inhibitor P55089 -01 but increased radioresistance of bystander cells . This study identifies P38398 , Q9BXW9 and Chk1 as potential targets for the modulation of radiation response in bystander cells . It adds to our understanding of the key molecular events propagating out-of-field effects of radiation and provides a rationale for the development of novel molecular targeted drugs for radiotherapy optimisation . Purification and characterization of platelet aggregation inhibitors from snake venoms . Proteins that inhibit glycoprotein ( GP ) IIb/IIIa mediated platelet aggregation have been purified from the venom of two snake species . A small platelet aggregation inhibitor ( p1.AI ) , multisquamatin ( Mr = 5,700 ) , was purified from Echis multisquamatus venom by hydrophobic interaction HPLC and two steps on C18 reverse phase HPLC . A larger p1.AI , contortrostatin ( Mr = 15,000 ) , was purified by a similar HPLC procedure from the venom of Agkistrodon contortrix contortrix . Both p1.AIs inhibit ADP-induced human , canine and rabbit platelet aggregation using platelet rich plasma ( PRP ) . Multisquamatin has an IC50 of 97 nM , 281 nM and 333 nM for human , canine and rabbit PRP , respectively . Contortrostatin has an IC50 of 49 nM , 120 nM and 1,150 nM for human , canine and rabbit PRP , respectively . In a competitive binding assay using 125I- DB00054 ( a monoclonal antibody to P08514 /IIIa that inhibits platelet aggregation ) both contortrostatin and multisquamatin demonstrated P08514 /IIIa specific binding to human and canine platelets . The IC50 for contortrostatin displacement of 7E3 binding to human and canine P08514 -/IIIa is 27 nM and 16 nM , respectively and for multisquamatin it is 3 nM and 63 nM , respectively . Our results indicate that both p1.AIs inhibit platelet aggregation by binding with high affinity to P08514 /IIIa . Increased activated protein C-protein C inhibitor complex levels in patients with pulmonary embolism . DB00055 ( P25054 ) -protein C inhibitor ( P05154 ) complex level was examined in 35 patients with acute pulmonary embolism ( PE ) and in 20 healthy volunteers . Thrombin-antithrombin III complex , plasmin alpha 2 plasmin inhibitor complex , and fibrin-D-dimer levels were significantly increased in the patients with PE compared to levels in healthy volunteers . Levels of plasminogen activator inhibitor-I , tissue type plasminogen activator , and P04275 antigens were also significantly increased in patients with PE . Plasma level of P25054 - P05154 complex was increased in most patients with PE and P25054 -alpha 1 antitrypsin complex level was increased in 13 patients . These complexes were not detected in healthy volunteers . These findings suggested that plasma protein C was activated in patients with PE , and that P05154 was the major inhibitor of P25054 generated in this condition . Thus , regulation of the protein C pathway might play an important role in the pathogenesis of PE . 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 . Lymphangioma involving the mandible : immunohistochemical expressions for the lymphatic proliferation . We report a case of lymphangioma involving oral mucosa and mandible of an elderly female . The surgical and radiological examinations indicated that the lymphangioma was mainly distributed in the labial mucosa tissue , but had gradually extended into the periosteum and intrabony space of mandible . Immunohistochemical staining was also performed using antiseras of alpha-smooth muscle actin ( alpha-SMA ) , P04275 ( P04275 ) , angiogenin , vascular endothelial growth factor ( P15692 ) , and proliferating cell nuclear antigen ( P12004 ) to elucidate the pathogenetic implications of the intraosseous lymphangioma . The present case of lymphangioma showed strong immunohistochemical reactivity of angiogenin and P04275 , while it showed weak reactions of P15692 and P12004 . The immunostaining of alpha-SMA disclosed an abnormally thinned and discontinuous smooth muscle layer in the lymphatics . Both the X-rays and histological examination showed that the lymphangioma lesion was gradually extending into the adjacent osteoporotic marrow space of mandible . Therefore , we believe that the present case of intraosseous lymphangioma , which showed the harmatomatous growth of the lymphatics into the marrow space of mandible , is closely related to osteoporotic changes of old age . 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 . 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 . 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 . Founder effect and estimation of the age of the French Gypsy mutation associated with Glanzmann thrombasthenia in Manouche families . The c.1544+1G > A substitution at the 5' splice donor site of intron 15 of the P08514 gene , called the French Gypsy mutation , causes Glanzmann thrombasthenia , an inherited hemorrhagic disorder transmitted as an autosomal recessive trait and characterized by an altered synthesis of the platelet αIIbβ3 integrin . So far , this mutation has only been found in affected individuals originating from French Manouche families , strongly suggesting a founder effect . Our goal was to investigate the origin of the French Gypsy mutation . We estimated the age of the mutation by a likelihood-based method that uses the length of the shared haplotypes among a set of patients . For this , we genotyped 23 individuals of Manouche origin ; consisting of 9 Glanzmann thrombasthenia patients homozygous for the French Gypsy mutation , 6 heterozygous carriers and 8 homozygous wild-type individuals . They were genotyped for four single-nucleotide polymorphisms using high-resolution melting curve analysis , and for two CA repeats in the P38398 and P10827 genes at chromosome 17 , using fragment analysis gels . We found that a haplotype of five polymorphic loci covering a 4-cM region was strongly associated with the French Gypsy mutation , suggesting a founder effect . The estimated age of this founder mutation was 300-400 years ( range 255-552 years ) . Thus , all carriers of the French Gypsy mutation c.1544+1G > A at intron 15 descended from a common ancestor 300-400 years ago . 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 . 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 . 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 . 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 . 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 . 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 ) 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 . 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 . 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 . 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 . 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 . 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 . 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 . Comparison of the effects of cytoprotective drugs on human plasma adrenocorticotropic hormone and cortisol levels with continual stress exposure . Cetraxate hydrochloride ( cetraxate ) , ecabet sodium ( ecabet ) , and sulpiride , which are cytoprotective drugs , have been used to treat peptic ulcers and acute or chronic gastritis . They are reported to improve mucosal blood flow in the stomach . One of the most important factors believed to cause gastric ulcers is mental and/or physiological stress . When people feel stress , the hypothalamo-pituitary-adrenal ( Q9Y251 ) axis is activated . Therefore , corticotropin-releasing hormone ( P06850 ) , adrenocorticotropic hormone ( DB01285 ) , and cortisol can be indicators of stress . We examined the effects of cetraxate , ecabet and sulpiride on the plasma levels of DB01285 and cortisol under stress conditions by repetitive blood sampling . Venous blood samples were taken before and 20-240 min after a single administration of the drugs or a placebo . A single dose of ecabet caused significant suppression of increases in plasma DB01285 -like immunoreactive substance ( IS ) levels at 90 to 120 min and cortisol levels at 240 min , compared with the response to placebo . DB00391 only suppressed increases in plasma cortisol levels at 180 to 240 min , compared with the response to placebo . A single dose of cetraxate had no effect on plasma DB01285 -IS and cortisol levels . Ecabet may have a modulatory effect on the Q9Y251 axis while sulpiride may have a partial modulatory effect on the Q9Y251 axis . These effects might be beneficial in stress-related disease . 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 . 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 . 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 . 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 . Inhibitory effect of chlorpromazine on O14788 -induced osteoclastogenesis in mouse bone marrow cells . Chlorpromazine ( CPZ ) , the first widely used phenothiazine tranquilizer , is shown to inhibit the action of intracellular calmodulin ( P62158 ) and bone resorption in vivo and in vitro . In this study , CPZ ( 0.63 - 10 µM ) dose-dependently inhibited the formation of tartrate-resistant acid phosphatase ( TRAP ) staining-positive osteoclast-like cells in mouse bone marrow cells ( BMCs ) treated with 1α,25(OH)(2)D(3) ( 10 nM ) or soluble receptor activator of nuclear factor-κB ligand ( s- O14788 ) ( 20 ng/ml ) . Expressions of mRNA for the nuclear factor of activated T-cells c1 ( O95644 ) , a key regulator of osteoclast differentiation ; dendritic cell-specific transmembrane protein ( Q9H295 ) , an essential protein for cell-cell fusion ; and characteristic markers of osteoclasts such as TRAP , cathepsin K , carbonic anhydrase II , and calcitonin receptor in BMCs were up-regulated by s- O14788 and decreased by the addition of CPZ ( 5 µM ) or the selective P62158 antagonist W7 , but not the inactive analog W5 . The general P62158 kinase ( CaMK ) inhibitor KN-93 and P62158 -dependent phosphatase calcineurin inhibitor FK-506 also inhibited s- O14788 -induced osteoclastogenesis . Phenothiazines such as CPZ , trifluoperazine ( TFPZ ) , and promethazine ( PMZ ) inhibited s- O14788 -induced osteoclast-like cell formation in mouse BMCs . Osteoclastogenesis inhibitory effects decreased in the order of TFPZ , CPZ , PMZ , depending on their anti- P62158 potency . These findings suggest that CPZ inhibits O14788 -induced osteoclastogenesis by its anti- P62158 action . Using ImageJ for the quantitative analysis of flow-based adhesion assays in real-time under physiologic flow conditions . This article intends to close the gap between the abundance of regular articles focusing on adhesive mechanisms of cells in a flow field and purely technical reports confined to the description of newly developed algorithms , not yet ready to be used by users without programming skills . A simple and robust method is presented for analysing raw videomicroscopic data of flow-based adhesion assays using the freely available public domain software ImageJ . We describe in detail the image processing routines used to rapidly and reliably evaluate the number of adherent and translocating platelets in videomicroscopic recordings . The depicted procedures were exemplified by analysing platelet interaction with immobilized P04275 and fibrinogen in flowing blood under physiological wall shear rates . Neutralizing GPIbalpha function reduced shear-dependent platelet translocation on P04275 and abolished firm platelet adhesion . DB00054 , Tirofiban and DB00063 completely inhibited P08514 /IIIa-dependent stable platelet deposition on fibrinogen . The presented method to analyse videomicroscopic recordings from flow-based adhesion assays offers the advantage of providing a simple and reliable way to quantify flow-based adhesion assays , which is completely based on ImageJ and can easily be applied to study adhesion mechanisms of cells in non-fluorescent modes without the need to deviate from the presented protocol . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Glycoprotein IIb/IIIa inhibition enhances platelet nitric oxide release . INTRODUCTION : Platelet aggregates form by fibrinogen binding to the membrane receptor glycoprotein IIb/IIIa ( P08514 /IIIa ) . While P08514 /IIIa inhibitors block fibrinogen-platelet binding , stimulation of other functionally important platelet receptors may still occur . Blocking the P08514 /IIIa receptor prevents platelet aggregation but not activation and the subsequent effect on other platelet pathways is largely unknown . MATERIALS AND METHODS : As activated platelets release reactive oxygen species that may influence thrombosis or vascular function , the effect of P08514 /IIIa inhibitors on the platelet release of nitric oxide ( NO ) and superoxide was determined using an electrochemical detector and luminescence , respectively . Location of relevant platelet proteins and the interaction between platelets and leukocytes in the presence or absence of P08514 /IIIa inhibition was determined . RESULTS : Although incubation with P08514 /IIIa inhibitors completely abolished platelet aggregation , stimulation dependent NO release was significantly enhanced . Superoxide is known to alter the bioavailability of NO , and its contribution to the P08514 /IIIa dependent increase in NO release was determined . In the presence of P08514 /IIIa inhibitors , platelet superoxide release was significantly decreased . Preincubation with P08514 /IIIa inhibitors also modified aggregation induced membrane translocation of the platelet proteins , endothelial NO synthase ( P29474 ) and NADPH oxidase ( p67phox and p47phox ) , known to contribute to the generation of NO and superoxide , respectively . In the presence of leukocytes , abciximab incubation led to enhanced NO release and attenuated superoxide generation . CONCLUSION : These observations suggest that the pharmacological effects of P08514 /IIIa antagonists on platelet function , apart from inhibition of aggregation , may contribute to their efficacy . Effect of glycoprotein IIb-IIIa receptor antagonism on platelet membrane glycoproteins after coronary stent placement . Platelet membrane glycoproteins play a crucial role in ischemic complications after coronary stenting . Glycoprotein IIb-IIIa blockade reduces adverse clinical events after angioplasty but is associated with rare but profound thrombocytopenia that might increase hemorrhagic complications . Changes in platelet membrane glycoproteins of patients with angina who underwent coronary stenting and were treated with the P08514 -IIIa antagonist abciximab ( n=20 ) or with heparin ( n=23 ) were studied . GPIb-IIIa receptor blockade and membrane glycoproteins were evaluated with immunological markers in venous blood samples taken before . 10 , 24 , 48 , 72 , and 96 h after initial treatment with either abciximab or heparin . Patients receiving abciximab therapy showed a rapid inhibition of binding of fluorochrome-conjugated mAb CD41 and c7E3 concomitant with a reduction in platelet aggregation which was restored in part in the days after termination of abciximab infusion . Induction of ligand-induced binding sites on P08514 -IIIa was increased in patients receiving abciximab . The expression of ligand-induced binding sites correlated inversely with platelet count . No significant change in platelet membrane markers were found in the heparin group . In vitro studies showed that abciximab induces ligand-induced binding sites on isolated platelets and on nuclear cells bearing recombinant P08514 -IIIa . DB00054 rapidly achieves P08514 -IIIa receptor blockade after coronary stent placement that might be beneficial in high-risk settings to bridge the delayed action of ticlopidine . Significant alterations of platelet membrane glycoproteins during P08514 -IIIa antagonism might contribute to development of acute profound thrombocytopenia . Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins with flow cytometry . Disintegrins are a group of snake venom peptides which inhibit human platelet aggregation by acting as glycoprotein IIb-IIIa ( P08514 -IIIa ) antagonists . They are cysteine-rich , DB00125 - DB00145 - DB00128 ( RGD ) -containing peptides , and bind to P08514 -IIIa complex on platelet membrane with a very high affinity ( Kd , 10(-7)-10(-8) M ) . In this study , we analyzed P08514 -IIIa complex on platelet membrane by flow cytometry using fluorescein isothiocyanate ( FITC ) -conjugated disintegrins as probes . Of these FITC-conjugated disintegrins , FITC-Rhodostomin is the most sensitive probe because Rhodostomin was conjugated with more FITC molecules than Trigramin and Halysin were . The binding fluorescence intensity of FITC-Trigramin ( FITC-Tg ) , FITC-Halysin ( FITC-Hy ) and FITC-Rhodostomin ( FITC-Rn ) was measured in both resting and ADP-activated platelets of diluted human platelet-rich plasma . The binding fluorescence of FITC-disintegrins was abolished by DB00974 and DB00054 , a monoclonal antibody against P08514 -IIIa . ADP markedly increased the fluorescence intensity of FITC-Tg and FITC-Hy bound on platelets especially when lower doses of these probes were used , whereas it had little effect on that of FITC-Rn . Therefore , FITC-Tg and FITC-Hy can be used for the detection of the activated platelets as noted by a higher ratio of fluorescence intensity ( approx. 2-4 ) between ADP-activated and resting platelets as compared with that ( approx. 1-1.3 ) in the case of FITC-Rn as the probe . The platelets from three patients with Glanzmann 's thrombasthenia were probed with FITC-disintegrins. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . Beta2-glycoprotein I binding to platelet microparticle membrane specifically reduces immunoreactivity of glycoproteins IIb/IIIa . We have investigated beta2-glycoprotein I ( beta2GPI ) binding to platelet-derived microparticles ( PMP ) and its effect on P08514 /IIIa . PMP were isolated from washed human platelets after stimulation with A23187 , and analyzed by surface plasmon resonance spectroscopy . Beta2GPI as well as activated protein C ( P25054 ) or annexin V bound to PMP-coated sensorchips , demonstrating exposure of anionic phospholipids on immobilized PMP . Beta2GPI binding was impaired by calcium and occurred in a concentration-dependent manner with apparent k(on) = 2.6 x 10(4) M(-1) s(-1) and k(off) = 4.4 x 10(-3) s(-1) , corresponding to a KD value of 1.7 x 10(-7) M . When analyzed by flow cytometry , the binding of certain mAbs specific for P08514 and/or P05106 was reduced in the presence of beta2GPI but not of P25054 or annexin V , whereas the binding of anti-GPIb or anti- P16109 mAbs , or of soluble fibrinogen remained unchanged . These results suggest a broad but specific influence of beta2GPI on P08514 /IIIa immunoreactivity , and indicate that beta2GPI may act as a modulator of P08514 /IIIa-dependent functions of PMP . 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 . 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 . 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 . 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 . [ Adjustment effects of Herba epimedii , Fructus ligustrilucidi on NO/ET , Q9Y251 axis in asthmatic rats ] . OBJECTIVE : To study the neuro-endocrine adjustment effects of Herba Epimedii and Fructus Ligustrilucidi on the asthmatic rats . METHOD : Rat asthma model was duplicated by OVA ( ovalbumin ) through sensitizing and challenging . Fifty male rats were randomly divided into normal group , model group , adjustment group of Herba Epimedii and Fructus Ligustrilucidi , Peibenfang group and Asimei capsule group . Investigating levels of ET ( Endothelin ) , NO , P35228 ( inducible NOS ) , and P29474 ( constitutive NOS ) in blood serum and BALF ( bronchoalveolar lavage fluid ) , O00230 ( corticotrophin ) in serum , DB01285 ( adrenocorticotropin hormone ) in plasma , CHR ( corticotropin release hormone ) in hypothalamus , protein expression of GCR ( glucocorticoid receptor ) in lung tissue . RESULT : The adjustment of Herba Epimedii and Fructus Ligustrilucidi could inhibit ET and NO content in BALF ( all P < 0.05 ) , decrease the level of P35228 in serum and BALF ( P < 0.01 or P < 0.05 ) , and increase the level of P29474 in serum and BALF ( P < 0.01 or P < 0.05 ) , raise the concentration of serum O00230 ( P < 0.01 ) , enhance the protein expression of GCR in lung tissue ( P < 0.05 ) . CONCLUSION : The preventive and therapeutic effect of Herba Epimedii and Fructus Ligustrilucidi on asthma relates to their adjustment effect on ET/NO and Q9Y251 axis . 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 . [ 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 . The functional study of human umbilical cord DB05914 harbouring angiotensin-converting enzyme 2 in rat acute lung ischemia-reperfusion injury model . Acute lung ischemia-reperfusion injury ( ALIRI ) is featured as non-specific alveolar damage , lung edema and hypoxemia , often occurring within 72 h after surgery . It is the leading cause for primary graft failure and mortality after lung surgery and transplantation . Here we aimed to find a more effective therapeutic approach to treat ALIRI . We evaluated the combinational effects of human umbilical cord DB05914 ( HUMSCs ) and angiotensin-converting enzyme 2 ( Q9BYF1 ) in the rat ALIRI model . HUMSCs were isolated for lentiviral- Q9BYF1 transfection . Fifty rats were randomly divided into five groups : sham surgery , physiological saline ( PS ) , Q9BYF1 , HUMSCs and HUMSCs- Q9BYF1 group . Several physiological , biochemical and histological indicators were examined and compared among the five groups , such as blood oxygen saturation ( Sat O2 % ) and right ventricular systolic blood pressure ( RVSBP ) , pulmonary morphology observations , several kinds of cell markers and the abundance of glutathione reductase ( GR ) , glutathione peroxidase ( GPX ) and NAD(P)H quinone oxidoreductase ( P15559 ) . Compared with HUMSCs and Q9BYF1 groups , HUMSCs- Q9BYF1 group showed lighter lung injuries , higher CD31 and P04275 expression ( endothelial cell surface markers ) , lower γ- P16104 ( DNA damage marker ) and P34810 ( inflammatory cell marker ) and higher anti-oxidants expression ( GR , GPX and P15559 ) . The results indicated that HUMSCs harbouring Q9BYF1 were more effective than either HUMSCs or Q9BYF1 alone in alleviating the ALIRI damages . The synergistic effects of HUMSCs and Q9BYF1 provide informative clues for mechanism study and therapeutic method development of ALIRI .

MH_train_1141

MH_train_1141

MH_train_1141

interacts_with DB06603?

multiple_choice

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 . Regulation of P60568 and Q02818 in mononuclear cells treated with acyl glucuronide of mycophenolic acid reveals effects independent of inosine monophosphate dehydrogenase inhibition . The aim of the present study was to investigate whether the acyl glucuronide of mycophenolic acid ( AcMPAG ) directly affects gene expression independent of guanosine ( G ) depletion . Human native mononuclear cells from healthy volunteers were studied . A concentration of 100 micromol/L ( 50 mg/L ) AcMPAG , which provided effective inhibition of cell proliferation according to dose-response curves , was selected for gene expression analysis on microarray , verified by quantitative real-time polymerase chain reaction on the LightCycler . Differentially regulated genes on the microarray were 114 inosine monophosphate dehydrogenase-independent genes involved in cell proliferation , signal transduction , chemokine stimulation , endocytosis , vesicle transport , cell adhesion , and cytoskeleton . For verification , 16 genes , which were directly or indirectly related to cell proliferation , were selected for quantitative real-time polymerase chain reaction . Q9BWG6 , Q9BTT0 , O43927 , P62158 , DKFZp451J0118 , P06753 , P60953 , P62258 , P19876 , P35241 , O95167 , and P00441 showed no significant difference between the studied groups ( P > 0.05 ) . P22362 gene expression was significantly regulated ( P < 0.05 ) only in the mononuclear cell group treated with AcMPAG , whereas P63104 gene expression was significantly regulated only in the group treated with AcMPAG in presence of G and 8-aminoguanosine . The difference of interleukin 2 ( P60568 ) and nucleobindin 1 ( Q02818 ) expression was significant between control and AcMPAG ( P < 0.05 ) however not significant between AcMPAG in presence and absence of G and 8-aminoguanosine ( P > 0.05 ) . The expression of interleukin 2 and nucleobindin 1 revealed an effect of AcMPAG on gene expression independent of inosine monophosphate dehydrogenase inhibition . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Neurotrophin p75 receptor ( p75NTR ) promotes endothelial cell apoptosis and inhibits angiogenesis : implications for diabetes-induced impaired neovascularization in ischemic limb muscles . Diabetes impairs endothelial function and reparative neovascularization . The p75 receptor of neurotrophins ( p75(NTR) ) , which is scarcely present in healthy endothelial cells ( ECs ) , becomes strongly expressed by capillary ECs after induction of peripheral ischemia in type-1 diabetic mice . Here , we show that gene transfer-induced p75(NTR) expression impairs the survival , proliferation , migration , and adhesion capacities of cultured ECs and endothelial progenitor cells ( EPCs ) and inhibits angiogenesis in vitro . Moreover , intramuscular p75(NTR) gene delivery impairs neovascularization and blood flow recovery in a mouse model of limb ischemia . These disturbed functions are associated with suppression of signaling mechanisms implicated in EC survival and angiogenesis . In fact , p75(NTR) depresses the P15692 /Akt/ P29474 /NO pathway and additionally reduces the mRNA levels of P05556 [ beta ( 1 ) integrin ] , O15392 ( survivin ) , O95997 ( securin ) and Q14119 . Diabetic mice , which typically show impaired postischemic muscular neovascularization and blood perfusion recovery , have these defects corrected by intramuscular gene transfer of a dominant negative mutant form of p75(NTR) . Collectively , our data newly demonstrate the antiangiogenic action of p75(NTR) and open new avenues for the therapeutic use of p75(NTR) inhibition to combat diabetes-induced microvascular liabilities . 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 . [ 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 . 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 . Functional P40763 deficiency compromises the generation of human T follicular helper cells . T follicular helper ( Tfh ) cells are critical for providing the necessary signals to induce differentiation of B cells into memory and Ab-secreting cells . Accordingly , it is important to identify the molecular requirements for Tfh cell development and function . We previously found that IL-12 mediates the differentiation of human P01730 (+) T cells to the Tfh lineage , because IL-12 induces naive human P01730 (+) T cells to acquire expression of Q9HBE4 , P41182 , Q9Y6W8 , and P32302 , which typify Tfh cells . We have now examined P01730 (+) T cells from patients deficient in IL-12Rβ1 , P29597 , P42224 , and P40763 to further explore the pathways involved in human Tfh cell differentiation . Although P42224 was dispensable , mutations in P42701 , P29597 , or P40763 compromised IL-12-induced expression of Q9HBE4 by human P01730 (+) T cells . Defective expression of Q9HBE4 by P40763 -deficient P01730 (+) T cells resulted in diminished B-cell helper activity in vitro . Importantly , mutations in P40763 , but not P42701 or P29597 , also reduced Tfh cell generation in vivo , evidenced by decreased circulating P01730 (+) P32302 (+) T cells . These results highlight the nonredundant role of P40763 in human Tfh cell differentiation and suggest that defective Tfh cell development and/or function contributes to the humoral defects observed in P40763 -deficient patients . The interplay between G protein-coupled receptor kinase 2 ( P25098 ) and histone deacetylase 6 ( Q9UBN7 ) at the crossroads of epithelial cell motility . G protein-coupled receptor kinase 2 ( P25098 ) is emerging as a key integrative node in cell migration control . In addition to its canonical role in the desensitization of G protein-coupled receptors involved in chemotaxis , novel recently identified P25098 substrates and interacting partners appear to mediate the P25098 -dependent modulation of diverse molecular processes involved in motility , such as gradient sensing , cell polarity or cytoskeletal reorganization . We have recently identified an interaction between P25098 and histone deacetylase 6 ( Q9UBN7 ) , a major cytoplasmic α-tubulin deacetylase involved in cell motility and adhesion . P25098 dynamically associates with and phosphorylates Q9UBN7 to stimulate its α-tubulin deacetylase activity at specific cellular localizations such as the leading edge of migrating cells , thus promoting local tubulin deacetylation and enhanced motility . This P25098 - Q9UBN7 functional interaction may have important implications in pathological contexts related to aberrant epithelial cell migration . 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 . 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 . Role of histone deacetylation in cell-specific expression of endothelial nitric-oxide synthase . Histone acetylation plays an important role in chromatin remodeling and gene expression . The molecular mechanisms involved in cell-specific expression of endothelial nitric-oxide synthase ( P29474 ) are not fully understood . In this study we investigated whether histone deacetylation was involved in repression of P29474 expression in non-endothelial cells . Induction of P29474 expression by histone deacetylase ( HDAC ) inhibitors trichostatin A ( P32119 ) and sodium butyrate was observed in all four different types of non-endothelial cells examined . Chromatin immunoprecipitation assays showed that the induction of P29474 expression by P32119 was accompanied by a remarkable increase of acetylation of histone H3 associated with the P29474 5'-flanking region in the non-endothelial cells . Moreover , DNA methylation-mediated repression of P29474 promoter activity was partially reversed by P32119 treatment , and combined treatment of P32119 and DB01262 ( AzadC ) synergistically induced P29474 expression in non-endothelial cells . The proximal Sp1 site is critical for basal activity of P29474 promoter . The induction of P29474 by inhibition of HDACs in non-endothelial cells , however , appeared not mediated by the changes in Sp1 DNA binding activity . We further showed that Sp1 bound to the endogenous P29474 promoter and associated with Q13547 in non-endothelial HeLa cells . Combined P32119 and AzadC treatment increased Sp1 binding to the endogenous P29474 promoter but decreased the association between Q13547 and Sp1 in HeLa cells . Our data suggest that Q13547 plays a critical role in P29474 repression , and the proximal Sp1 site may serve a key target for HDCA1-mediated P29474 repression in non-endothelial cells . 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 . 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 . 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 . 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 . Breed effect on early cytokine mRNA expression in spleen and cecum of chickens with and without Salmonella enteritidis infection . We examined mRNA expression of 11 genes : Q16611 , Bcl-x , Interferon [ IFN ] -gamma , Interleukin [ IL ] -1beta , P05231 , P22301 , IL-12alpha , IL-12beta , Q14116 , CXCLi2 [ P10145 / Q13111 ] , and a MIP family chemokine , CCLi2 , in the spleen and cecum of day-old chicks after oral inoculation with Salmonella enteritidis ( SE ) or medium . Three distinct chicken breeds ( broiler , Fayoumi , and Leghorn ) were evaluated for mRNA expression levels at 2 and 18h post-inoculation using quantitative reverse transcriptase-polymerase chain reaction ( RT-PCR ) . SE exposure significantly increased splenic Q14116 and P01579 expression . Breed effect was significant ( P < 0.05 ) for CXCLi2 , P22301 , IL-12alpha , and CCLi2 mRNA expression in the spleen , and for IL-12alpha , IL-12beta , Q14116 , and CCLi2 mRNA expression in the cecum . Generally , mRNA expression levels were higher in the spleen , and lower in the cecum , of Leghorns versus broilers . These results support a role for breed genetics influencing cytokine mRNA expression in young chickens and may potentially explain some generalized immune response differences between breeds . P11021 / P11021 / P11021 /Dna K is a universal therapeutic target for human disease . The chaperone P11021 /Dna K is conserved throughout evolution down to prokaryotes . The P11021 inhibitor OSU-03012 ( AR-12 ) interacted with sildenafil ( Viagra ) or tadalafil ( DB00820 ) to rapidly reduce P11021 levels in eukaryotes and as a single agent reduce Dna K levels in prokaryotes . Similar data with the drug combination were obtained for : HSP70 , HSP90 , P14625 , P30101 , HSP27 , P25685 and HSP60 . OSU-03012/sildenafil treatment killed brain cancer stem cells and decreased the expression of : O15118 and TIM1 ; P11279 ; and NTCP1 , receptors for Ebola/Marburg/Hepatitis A , Lassa fever , and Hepatitis B viruses , respectively . Pre-treatment with OSU-03012/sildenafil reduced expression of the coxsakie and adenovirus receptor in parallel with it also reducing the ability of a serotype 5 adenovirus or coxsakie virus B4 to infect and to reproduce . Similar data were obtained using Chikungunya , Mumps , Measles , Rubella , RSV , CMV , and Influenza viruses . OSU-03012 as a single agent at clinically relevant concentrations killed laboratory generated antibiotic resistant E. coli and clinical isolate multi-drug resistant N. gonorrhoeae and MRSE which was in bacteria associated with reduced Dna K and Q9UIJ5 A expression . The O76074 inhibitors sildenafil or tadalafil enhanced OSU-03012 killing in N. gonorrhoeae and MRSE and low marginally toxic doses of OSU-03012 could restore bacterial sensitivity in N. gonorrhoeae to multiple antibiotics . Thus , Dna K and bacterial phosphodiesterases are novel antibiotic targets , and inhibition of P11021 is of therapeutic utility for cancer and also for bacterial and viral infections . 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 . Q14116 activates P40763 in the natural killer cell line 92 , augments cytotoxic activity , and mediates P01579 production by the stress kinase p38 and by the extracellular regulated kinases p44erk-1 and p42erk-21 . Q14116 is a regulator of NK cell function which utilizes the serine-threonine IL-1R-associated kinase signal transduction pathway and may activate additional not yet characterized signaling pathways . Here we evaluated Q14116 -mediated signal transduction using the human NK cell line NK92 as a model . NK92 cells were shown by RT-PCR to express all three Q14116 receptor chains ( IL-18R , accessory protein-like chain , Q14116 -binding protein ) . Stimulation by Q14116 strongly enhanced tyrosine phosphorylation of P40763 and of the mitogen-activated protein kinases ( MAPK ) p44erk-1and p42erk-2 . In contrast , P42229 was not activated . The cytolytic activity of NK92 against K562 target cells , which was augmented in a dose-dependent manner by Q14116 in the presence of trace amounts of P60568 , was suppressed by the specific inhibitors of MAPK pathways ( PD098059 and SB203580 ) . Similarly , the stimulatory effect of Q14116 on P01579 protein production , given in conjunction with P60568 , was counteracted by inhibition of MAPK . Q14116 alone failed to stimulate P01579 protein production despite inducing expression of P01579 mRNA . P60568 alone stimulated neither P01579 mRNA expression nor P01579 protein production . Q14116 did not stimulate proliferation of NK92 cells , either alone or in combination with P60568 or IL-12 . Inhibition of the MAPK pathway did not significantly alter the P60568 - and IL-12-induced proliferation of NK92 cells , whereas the Janus kinase/ P35610 pathway inhibitor AG490 strongly suppressed proliferation . MAPK activation appears to play a prominent role in Q14116 signaling , being involved in transcription and translation of Q14116 -induced P01579 mRNA and Q14116 -induced cytolytic effects . In contrast , proliferation of NK92 cells is not affected by MAPK p44erk-1 and p42erk-2 . Effect of 27nt small RNA on endothelial nitric-oxide synthase expression . We have reported previously that the 27nt repeat polymorphism in endothelial nitric-oxide synthase ( P29474 ) intron 4 -- a source of 27nt small RNA -- inhibits P29474 expression . In the current study , we have investigated how 27nt small RNA suppresses P29474 expression . Using a chromatin immunoprecipitation assay , we examined histone acetylation in the 27nt repeat element of P29474 intron 4 , the promoter region up to -1486 bp , and the 5' enhancer region ( -4583/-4223bp ) in human aortic endothelial cells ( HAECs ) treated with 27nt RNA duplex . 27nt RNA duplex induced hyperacetylation in H3 ( lysine8 , 12 , and 23 ) and H4 ( lysine 9 and 12 ) at the 27nt repeat element , which then interacted with nuclear actin , histone deacetylase 3 ( O15379 ) , and NonO proteins . In contrast , the histone H3 and H4 became hypoacetylated at the P29474 core promoter . HAECs treated with 27nt RNA duplex had reduced P29474 expression , but treatment with either O15379 small interfering RNA or NonO siRNA significantly attenuated the 27nt small RNA-induced suppression . We further found that 27nt small RNA induced DNA methylation in a region approximately 750nt upstream of the intron 4 repeats , and a methyltransferase inhibitor reversed the effect on methylation and P29474 expression . Our study demonstrates that 27nt small RNA may suppress P29474 expression by altering histone acetylation and DNA methylation in regions adjacent to the 27nt repeat element and core promoter . Greglist : a database listing potential G-quadruplex regulated genes . The double helix is a conformation that genomic DNA usually assumes ; under certain conditions , however , guanine-rich DNA sequences can form a four-stranded structure , G-quadruplex , which is found to play a role in regulating gene expression . Indeed , it has been demonstrated that the G-quadruplex formed in the c-MYC promoter suppresses its transcriptional activity . Recent studies suggest that G-quadruplex motifs ( GQMs ) are enriched in human gene promoters . To facilitate the research of G-quadruplex , we have constructed Greglist , a database listing potentially G-quadruplex regulated genes . Greglist harbors genes that contain promoter GQMs from genomes of various species , including humans , mice , rats and chickens . Many important genes are found to contain previously unreported promoter GQMs , such as Q13315 , Q92934 , P31749 , LEPR , P25874 , P02649 , O94907 , P19544 , P30291 , P04628 and O15516 . Furthermore , we find that not only protein coding genes , 126 human microRNAs also contain promoter GQMs . Greglist therefore provides candidates for further studying G-quadruplex functions and is freely available at http://tubic.tju.edu.cn/greglist . 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 . 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 . Tenocyte apoptosis in the torn rotator cuff : a primary or secondary pathological event ? Little information exists on the contribution of apoptosis to pathological tendon changes in rotator cuff tendinopathy . The purpose of this study was to quantitate the rate of tenocyte apoptosis in torn supraspinatus tendons and in the matched intact subscapularis and to examine the potential relation between apoptotic index ( AI ) and tendon pathology . In addition , the authors examined tenocyte density , proliferation rate and p53 gene expression patterns to gain further insight into relevant pathological mechanisms in the torn suprapinatus . 15 torn supraspinatus tendons with matched intact subscapularis tendon samples and 10 reference subscapularis samples were collected . Immunohistochemistry was used to define the AI ( P08709 -26 ) , proliferation rate ( Ki67 ) and presence of p53 ( M7001 ) . Tendon degeneration was evaluated according to the Bonar scale . Expression of p53 and relevant genes ( n=84 ) was examined on a subset of samples using microfluidic arrays . The AI was significantly increased in torn supraspinatus tendon and matched subscapularis tendon ( R² =0.5742 ; p=0.0005 ) . Cell density and proliferation rate were also elevated in torn supraspinatus compared with reference subscapularis tendons ( p < 0.05 ) . A significant increase in p53 occurred specifically in torn supraspinatus tendon ( p < 0.05 ) , and several genes encoding p53-inhibiting proteins were downregulated in association , including Q13547 ( p < 0.05 ) , O15151 ( p < 0.001 ) and O15297 ( p < 0.05 ) . Our results suggest that tenocyte apoptosis results from more than one mechanism in the injured rotator cuff , including both intrinsic factors related specifically to the torn supraspinatus tendon , as well as a more generalised effect on the adjacent subscapularis tendon . Antiangiogenic activity of the endocannabinoid anandamide : correlation to its tumor-suppressor efficacy . Endocannabinoids are now emerging as suppressors of key cell-signaling pathways involved in cancer cell growth , invasion , and metastasis . We have previously observed that the metabolically stable anandamide analog , 2-methyl-2'-F-anandamide ( DB00134 -F-AEA ) can inhibit the growth of thyroid cancer in vivo . Our hypothesis was that the anti-tumor effect observed could be at least in part ascribed to inhibition of neo-angiogenesis . Therefore , the aim of this study was to assess the anti-angiogenic activity of DB00134 -F-AEA , to investigate the molecular mechanisms underlying this effect and whether DB00134 -F-AEA could antagonize tumor-induced endothelial cell sprouting . We show that DB00134 -F-AEA inhibited P09038 -stimulated endothelial cell proliferation , in a dose-dependent manner , and also induced apoptosis , both effects reliant on cannabinoid P21554 receptor stimulation . Analyzing the signaling pathways implicated in angiogenesis , we observed that the P09038 -induced P29323 phosphorylation was antagonized by DB00134 -F-AEA , and we found that p38 MAPK was involved in DB00134 -F-AEA-induced apoptosis . Moreover , DB00134 -F-AEA was able to inhibit bi-dimensional capillary-like tube formation and activity of matrix metalloprotease P08253 , a major matrix degrading enzyme . Importantly , we demonstrated that DB00134 -F-AEA is also functional in vivo since it inhibited angiogenesis in the chick chorioallantoic neovascularization model . Finally , DB00134 -F-AEA inhibited tumor-induced angiogenesis in a three-dimensional model of endothelial and thyroid tumor cell ( KiMol ) spheroids co-cultures in different 3-D polymeric matrices that resemble tumor microenvironment and architecture . Thus , our results suggest that anandamide could be involved in the control of cancer growth targeting both tumor cell proliferation and the angiogenic stimulation of the vasculature . 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 . B7h triggering inhibits umbilical vascular endothelial cell adhesiveness to tumor cell lines and polymorphonuclear cells . Vascular endothelial cells ( ECs ) are key players in leukocyte recruitment into tissues and metastatic dissemination of tumor cells . ECs express B7h , which is the ligand of the Q9Y6W8 T cell costimulatory molecule . The aim of this work was to assess the effect of B7h triggering by a soluble form of Q9Y6W8 ( Q9Y6W8 -Fc ) on the adhesion of colon carcinoma cell lines to HUVECs . We found that B7h triggering inhibited HUVEC adhesiveness to HT29 and DLD1 cells ( by 50 and 35 % , respectively ) but not to HCT116 cells . The effect was dependent on the Q9Y6W8 -Fc dose and was detectable as early as 30 min after treatment and was still present after 24 h . It was inhibited by soluble anti- Q9Y6W8 reagents ( mAb and B7h-Fc ) and silencing of B7h on HUVECs , and it was not displayed by an F119S mutated form of Q9Y6W8 -Fc that does not bind B7h . HUVEC treatment with Q9Y6W8 -Fc did not modulate expression of adhesion molecules and cytokines , but it substantially downmodulated P29323 phosphorylation induced by P16581 triggering or osteopontin , which may influence HUVEC adhesiveness . Moreover , HUVEC treatment with Q9Y6W8 -Fc also inhibited adhesion of polymorphonuclear cells and several tumor cell lines from different origins . Therefore , the B7h- Q9Y6W8 interaction may modulate spreading of cancer metastases and recruitment of polymorphonuclear cells in inflammatory sites , which opens a view on the use of Q9Y6W8 -Fc as an immunomodulatory drug . Increased expression of chemerin in squamous esophageal cancer myofibroblasts and role in recruitment of mesenchymal stromal cells . Stromal cells such as myofibroblasts influence tumor progression . The mechanisms are unclear but may involve effects on both tumor cells and recruitment of bone marrow-derived mesenchymal stromal cells ( MSCs ) which then colonize tumors . Using iTRAQ and LC-MS/MS we identified the adipokine , chemerin , as overexpressed in esophageal squamous cancer associated myofibroblasts ( CAMs ) compared with adjacent tissue myofibroblasts ( ATMs ) . The chemerin receptor , ChemR23 , is expressed by MSCs . Conditioned media ( CM ) from CAMs significantly increased O60682 cell migration compared to Q13315 -CM ; the action of P62158 -CM was significantly reduced by chemerin-neutralising antibody , pretreatment of CAMs with chemerin siRNA , pretreatment of MSCs with ChemR23 siRNA , and by a ChemR23 receptor antagonist , CCX832 . Stimulation of MSCs by chemerin increased phosphorylation of Q8NFH3 /44 , p38 and JNK-II kinases and inhibitors of these kinases and PKC reversed chemerin-stimulated O60682 migration . Q99969 stimulation of MSCs also induced expression and secretion of macrophage inhibitory factor ( MIF ) that tended to restrict migratory responses to low concentrations of chemerin but not higher concentrations . In a xenograft model consisting of OE21 esophageal cancer cells and CAMs , homing of MSCs administered i.v. was inhibited by CCX832 . Thus , chemerin secreted from esophageal cancer myofibroblasts is a potential chemoattractant for MSCs and its inhibition may delay tumor progression . 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 . 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 . 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 . Production and characterization of monoclonal antibodies against urea derivatives . A panel of monoclonal antibodies was generated against the urea-based hapten N-(2-N-chloroacetylaminobenzyl)-N'-4-chlorophenylurea as a tool for building up sensitive immune assays to detect urea derivatives and to screen them for catalytic antibodies ( Abs ) . Eleven hybridomas were obtained that produced Abs reactive to the hapten . All Abs were of IgG class . Cross reactivities of the Abs to different haptens were examined , especially to a possible transition-state analog . Only four of the hybridomas ( R2-DA10/ P08709 , R2-GE7/H2 , R2- P25786 /A5 , R2- Q9UBN7 / P08709 ) produced Abs crossreactive with the transition-state analog . From the 11 hybridomas , hybridoma B76-BF5 was chosen for further characterization . Compared to the other Abs , B76-BF5 showed the strongest binding and had a rather restricted specificity . These Abs could be used to build up a sensitive enzyme immunoassay for the detection of the hapten . All Abs were screened for crossreactivity with the pesticides monuron and diuron . No reactivity could be detected . In addition , the nucleotide sequences of the variable light and heavy chain genes of the similarly reactive Abs B76-BF5 , B76-BB3 , R2-DA10/ P08709 , and R2-GA6/ P46379 were determined to clarify whether structure and binding specificity of these Abs showed any correlation . 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 . 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 . 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 . 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 . Interleukin 23 regulates proliferation of lung cancer cells in a concentration-dependent way in association with the interleukin-23 receptor . A proinflammatory cytokine , interleukin 23 ( IL-23 ) , plays a role in tumor progression by inducing inflammation in the tumor microenvironment , although there is debate about its role in carcinogenesis . Direct effects of IL-23 on tumor cells have been reported rarely , and contradictory effects have been observed . Here , we studied such effects of IL-23 in lung cancer cells in vitro and in vivo and explored the underlying mechanism . We found Q5VWK5 expression in tissues from lung adenocarcinoma and small cell carcinoma but not in lung squamous cell carcinoma tissue . Interestingly , different concentrations of IL-23 had opposite effects in the same types of cells . We confirmed that the different effects could be explained by differences in binding to the Q5VWK5 ( subunits IL-23r and IL-12Rβ1 ) . Low concentrations of IL-23 promoted the proliferation of Q5VWK5 -positive A549 and P08709 -1 lung cancer cells by binding to IL-23r , whereas high concentrations of IL-23 inhibited proliferation of these cells by binding to both IL-23r and IL-12Rβ1 . In contrast , IL-23 had no effect on Q5VWK5 -negative SK-MES-1 cells . IL-23 regulated the growth of human lung cancer cells through its effects on P40763 expression and phosphorylation in a concentration-dependent way ; the Ki-67 gene was involved in these processes . Our findings demonstrate for the first time that IL-23 affects the proliferation of Q5VWK5 -positive lung cancer cells and that this effect is dependent on the IL-23 concentration . This can explain at least part of the inconsistent reports on the role of IL-23 in the progression of carcinogenesis . The mechanism of how anti- Q14116 prevents concanavalin-A-induced hepatic fibrosis on a mouse model . BACKGROUND : The administration of concanavalin A ( ConA ) induces severe hepatic fibrosis in mice . P01375 -alpha ( P01375 ) , interferon-gamma ( P01579 ) and interleukin 4 ( P05112 ) were the key cytokines involved in the process . The aim of this research was to explore the effects and the mechanisms of Q14116 and anti- Q14116 on hepatic fibrosis in a ConA induced hepatic fibrosis model in BABL-C mice . MATERIALS AND METHODS : One hundred eighty BABL-C mice were randomly divided into five groups ( Group a , b , c , d , e ) . The mice were administered saline , immunoglobulin G , ConA , Q14116 + ConA , Anti- Q14116 + ConA , respectively . At 1 , 7 , 14 , 21 wk , the levels of serum alanine aminotransferase , P01375 , P01579 , P05112 , matrix metalloproteinase ( MMP ) -2-RNA , and tissue inhibitor of metalloproteinase-1-mRNA were measured . RESULTS : The levels of serum P01375 and P01579 detected in the Q14116 + ConA group was higher than in the anti- Q14116 + ConA group ( P < 0.05 ) . Similarly , the levels of P08253 -RNA and tissue inhibitor of metalloproteinase-1-mRNA expressed in Q14116 + ConA group was higher than in the anti- Q14116 + ConA group ( P < 0.05 ) . A majority of these cytokines was secreted by P01730 (+)T cells . CONCLUSIONS : The immunological response to hepatic fibrosis by repeated injection of ConA in the mouse model was aggravated by Q14116 and blocked by anti- Q14116 . 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 . 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 . [ Down-regulation of P11021 Enhances Chemotherapy Sensitivity to DB00773 in Lung Adenocarcinoma. ] . BACKGROUND : P11021 , a member of GRPs , plays a critical role in chemotherapy resistance in some cancers . To investigate the relationship between the expression of P11021 and resistance to anti-cancer drug DB00773 in vitro in lung adenocarcinoma P08709 -1 cell line . METHODS : P08709 -1 cells were divided into three groups : BAPTA-AM-treated group , A23187-treated group and the control group . RT-PCR and immunofluorescence were used to analyze the expression of P11021 at both mRNA and protein levels , respectively . Cell apoptosis was analyzed by flow cytometry in order to evaluate the therapeutic sensitivity to DB00773 . RESULTS : The expression of P11021 at both protein and mRNA levels in the BAPTA-AM-treated cells dramatically decreased as compared to that of both A23187-treated and control groups . After treatment by DB00773 , the percentages of apoptotic cells were 10.84+/-0.86 , 6.85+/-0.20 , 4.95+/-0.19 in BAPTA-M-treated group , the control group and A23187-treated group , respectively . CONCLUSIONS : BAPTA-AM is highly effective in the inhibition of P11021 , down-regulation of P11021 can significantly increase the sensitivity of adenocacinoma lung cancer to DB00773 . All these suggest that inhibition of the expression of P11021 by chemicals such as BAPTA-AM or anti-sense RNA may be a new therapeutic strategies to lung cancer . DB11320 reduces susceptibility to natural killer cells via down-regulation of P26718 ligands on human monocytic leukaemia THP-1 cells . Natural killer ( NK ) group 2D ( P26718 ) is a key activating receptor expressed on NK cells , whose interaction with ligands on target cells plays an important role in tumorigenesis . However , the effect of histamine on P26718 ligands on tumour cells is unclear . Here we showed that human monocytic leukaemia THP-1 cells constitutively express MHC class I-related chain A ( Q29983 ) and Q9BZM6 on their surface , and incubation with histamine reduced the expression in a dose-dependent and time-dependent manner as assessed by flow cytometry . Interferon-γ augmented the surface expression of the P26718 ligands , and this augmentation was significantly attenuated by histamine . The histamine H1 receptor ( P35367 ) agonist 2-pyridylethylamine and P25021 agonist dimaprit down-regulated the expression of P26718 ligands , and activation of P35367 and P25021 signalling by A23187 and forskolin , respectively , had the same effect , indicating that the histamine-induced down-regulation of P26718 ligands is mediated by P35367 and P25021 . Quantitative reverse transcription-PCR showed that mRNA levels of the P26718 ligands and relevant microRNAs were not significantly changed by histamine . DB11320 down-regulated the surface expression of endoplasmic reticulum protein 5 , and inhibition of matrix metalloproteinases did not impair this down-regulation , indicating that proteolytic shedding was not involved . Instead , pharmacological inhibition of protein transport and proteasome abrogated it , and histamine enhanced ubiquitination of Q29983 . Furthermore , histamine treatment significantly reduced susceptibility to NK cell-mediated cytotoxicity . These results suggest that histamine down-regulates P26718 ligands through the activation of an P35367 - and P25021 -mediated ubiquitin-proteasome pathway and consequently reduces susceptibility to NK cells . 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 . 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 . O15379 represses the expression of P26718 ligands ULBPs in epithelial tumour cells : potential implications for the immunosurveillance of cancer . The expression of the P26718 ligands on cancer cells leads to their recognition and elimination by host immune responses mediated by natural killer and T cells . UL16-binding proteins ( ULBPs ) are P26718 ligands , which are scarcely expressed in epithelial tumours , favouring their evasion from the immune system . Herein , we investigated the epigenetic mechanisms underlying the repression of ULBPs in epithelial cancer cells . We show that Q9BZM6 -3 expression is increased in tumour cells after exposure to the inhibitor of histone deacetylases ( HDACs ) trichostatin A ( P32119 ) , which enhances the natural killer cell-mediated cytotoxicity of HeLa cells . Our experiments showed that the transcription factor Sp3 is crucial in the activation of the Q9BZM6 promoter by P32119 . Furthermore , by small interfering RNA-mediated knockdown and overexpression of Q13547 -3 , we showed that O15379 is a repressor of ULBPs expression in epithelial cancer cells . Remarkably , P32119 treatment caused the complete release of O15379 from the Q9BZM6 -3 promoters . O15379 is recruited to the Q9BZM6 promoter through its interaction with Sp3 and P32119 treatment interfered with this association . Together , we describe a new mechanism by which cancer cells may evade the immune response through the epigenetic modulation of the ULBPs expression and provide a model in which HDAC inhibitors may favour the elimination of transformed cells by increasing the immunogenicity of epithelial tumours . 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 .

MH_train_1142

MH_train_1142

MH_train_1142

interacts_with DB00243?

multiple_choice

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 . 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 . 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 . Mucin secretion induced by titanium dioxide nanoparticles . Nanoparticle ( NP ) exposure has been closely associated with the exacerbation and pathophysiology of many respiratory diseases such as Chronic Obstructive Pulmonary Disease ( P48444 ) and asthma . Mucus hypersecretion and accumulation in the airway are major clinical manifestations commonly found in these diseases . Among a broad spectrum of NPs , titanium dioxide ( TiO(2) ) , one of the PM10 components , is widely utilized in the nanoindustry for manufacturing and processing of various commercial products . Although TiO(2) NPs have been shown to induce cellular nanotoxicity and emphysema-like symptoms , whether TiO(2) NPs can directly induce mucus secretion from airway cells is currently unknown . Herein , we showed that TiO(2) NPs ( < 75 nm ) can directly stimulate mucin secretion from human bronchial ChaGo- P04264 epithelial cells via a Ca(2+) signaling mediated pathway . The amount of mucin secreted was quantified with enzyme-linked lectin assay ( ELLA ) . The corresponding changes in cytosolic Ca(2+) concentration were monitored with Rhod-2 , a fluorescent Ca(2+) dye . We found that TiO(2) NP-evoked mucin secretion was a function of increasing intracellular Ca(2+) concentration resulting from an extracellular Ca(2+) influx via membrane Ca(2+) channels and cytosolic ER Ca(2+) release . The calcium-induced calcium release ( CICR ) mechanism played a major role in further amplifying the intracellular Ca(2+) signal and in sustaining a cytosolic Ca(2+) increase . This study provides a potential mechanistic link between airborne NPs and the pathoetiology of pulmonary diseases involving mucus hypersecretion . 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 . Overexpression of cytochrome P450 4F2 in mice increases 20-hydroxyeicosatetraenoic acid production and arterial blood pressure . P78329 ( P78329 ) activity is thought to be a factor in the pathogenesis of hypertension through its bioactive metabolite 20-hydroxyeicosatetraenoic acid ( 20-HETE ) . We previously found that a gain-in-function P78329 variant in a Chinese cohort was associated with elevated urinary 20-HETE and hypertension . To further explore this association we generated a transgenic mouse model expressing P78329 driven by a modified mouse kidney androgen-regulated protein promoter . This heterologous promoter regulated the expression of luciferase and his-tagged P78329 in transfected P29320 293 cells . In the kidney of transgenic mice , P78329 was localized to renal proximal tubule epithelia and was expressed at a higher level than in control mice , leading to increased urinary 20-HETE excretion . Assessment of P78329 activity by an arachidonic acid hydroxylation assay showed that 20-HETE production was significantly higher in kidney microsomes of transgenic mice compared to control mice , as was their systolic blood pressure . There was a positive correlation of blood pressure with urinary 20-HETE levels . Our results show that increased expression of P78329 in mice enhanced 20-HETE production and elevated blood pressure . 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 . 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 . Regulation of fibrillin-1 gene expression by Sp1 . Mutations in the fibrillin-1 gene ( P35555 ) cause Marfan Syndrome ( MFS ) , a hereditary disorder of connective tissue . The transcription of P35555 has been reported to be driven by a short ultraconserved region ( SUPR ) in the 5' untranslated exon A of P35555 , but the nature of other factors involved in P35555 gene regulation has not been clarified . In this study , we characterized the transcription factors involved in P35555 gene regulation . The results show that Sp1 protein binds to two putative binding sites in the promoter of P35555 . Overexpression of Sp1 resulted in a significant increase in both promoter activity and P35555 mRNA level in P29320 293 cells , whereas inhibition or knockdown of Sp1 decreased P35555 gene expression . In addition , we found that Poly [ ADP-ribose ] polymerase 1 ( P09874 ) binds to the palindromic sequence TCTCGCGAGA in the ultraconserved region of the P35555 promoter and that the regulation of P35555 expression by P09874 is dependent on Sp1 . These results indicate that both Sp1 and P09874 contribute to P35555 gene expression . These observations add to our understanding of the transcriptional regulation of P35555 gene expression . Inhibition of poly(ADP-ribose)polymerase does not affect the recombination events in CHO xrs6 and wild type cells . Activation of poly ( ADP-ribose ) polymerase -1 ( P09874 ) is an early DNA damage response event that , together with phosphorylation of p53 , prompts various cellular functions important in the maintenance of the genome stability . In mammalian cells , DSB are repaired by nonhomologous end-joining ( NHEJ ) and by homologous recombination ( HR ) . To investigate the role of P09874 in HR , CHO- P04264 wild type and xrs-6 mutant cell line were transfected with pLrec plasmids which carry two nonfunctional copies of the beta-galactosidase ( lacZ ) gene in a tandem array . In result of HR they can give rise to a functional copy of beta-galactosidase . To test whether P09874 affects the frequency of spontaneous and induced recombination repair , we treated CHO- P04264 and xrs6 clones carrying chromosomally integrated pLrec with the P09874 inhibitor 3-aminobenzamide ( 3AB ) . Our results show that the spontaneous homologous intrachromosomal recombination frequency between the two lacZ copies was almost two orders of magnitude higher in xrs6 cells than in CHO- P04264 cells , but that it was not affected by 3AB treatment . Induction of DNA damage by irradiation or electroporation of restriction enzymes did not significantly increase the recombination frequency . Furthermore , in both the cell lines , the effect of P09874 inhibition on DSB repair was examined using the neutral comet assay . There was no effect of 3AB treatment on DSB rejoining after 10 Gy irradiation . The results presented support the conclusion that P09874 is not directly involved in HR . 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 ' . Development of a cell-based assay for the detection of anti-aquaporin 1 antibodies in neuromyelitis optica spectrum disorders . OBJECTIVE : To develop a cell-based assay ( CBA ) to detect aquaporin 1 ( P29972 ) antibodies and determine sensitivity/specificity in patients with neuromyelitis optica ( NMO ) spectrum disorders . METHODS : A P29320 -293T transfected cell model expressing P29972 was established and detected to be serum P29972 antibodies . RESULTS : P29972 antibodies were present in 73/98 ( 74.5 % ) P55087 antibody-positive patients . Some P55087 antibody-negative patients were also P29972 antibody-positive . Test sensitivity was 74.5 % in 98 P55087 antibody-positive patients . Test specificity was 79.6 % in 67 multiple sclerosis ( MS ) patients and 31 controls . CONCLUSION : A sensitive and simple CBA was developed to detect serum P29972 antibodies . P29972 antibodies were mainly present in NMO and its high-risk syndrome , but also in some MS patients . 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 . 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 . 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 . The role of transmembrane domain 2 in cation transport by the Na-K-Cl cotransporter . The human and shark Na-K-Cl cotransporters ( NKCC ) , although 74 % identical in amino acid sequence , exhibit marked differences in ion transport and bumetanide binding . We have utilized shark-human chimeras of P55011 to search for regions that confer the kinetic differences . Two chimeras ( hs3.1 and its reverse sh3.1 ) with a junction point located at the beginning of the third transmembrane domain were examined after stable transfection in P29320 -293 cells . Each carried out bumetanide-sensitive 86Rb influx with cation affinities intermediate between shark and human cotransporters . In conjunction with the previous finding that the N and C termini are not responsible for differences in ion transport , the current observations identify the second transmembrane domain as playing an important role . Site-specific mutagenesis of two pairs of residues in this domain revealed that one pair is indeed involved in the difference in Na affinity , and a second pair is involved in the difference in P06400 affinity . Substitution of the same residues with corresponding residues from Q13621 or the P55017 resulted in cation affinity changes , consistent with the hypothesis that alternative splicing of transmembrane domain 2 endows different versions of Q13621 with unique kinetic behaviors . None of the changes in transmembrane domain 2 was found to substantially affect Km(Cl) , demonstrating that the affinity difference for Cl is specified by the region beyond predicted transmembrane domain 3 . Finally , unlike Cl , bumetanide binding was strongly affected by shark-human replacement of transmembrane domain 2 , indicating that the bumetanide-binding site is not the same as the Cl-binding site . The establishment and characterization of cell lines stably expressing human P13010 tagged with enhanced green fluorescent protein . The Ku protein is a complex of two subunits , P12956 and P13010 , and it plays a role in multiple nuclear processes , e.g. , nonhomologous DNA-end-joining ( NHEJ ) , chromosome maintenance , and transcription regulation . On the other hand , several studies have reported a cytoplasmic or cell surface localization of Ku in various cell types . The mechanism underlying the regulation of all the diverse functions of Ku is still unclear , though the mechanism that regulates the nuclear localization of P12956 and P13010 appears to play , at least in part , a key role in regulating the physiological function of Ku . In this study , we generated cell lines expressing the human P13010 tagged with the green fluorescent protein ( GFP ) color variants in P13010 -deficient cells , i.e. , xrs-6 derived from CHO- P04264 . Although P12956 , as well as P13010 , was undetectable in xrs-6 cells , it was seen in these transformants at a level similar to the level of CHO- P04264 . Furthermore , etoposide- and radiosensitive phenotype of xrs-6 cells were corrected by an introduction of the tagged P13010 . Moreover , the tagged P13010 suppressed apoptosis triggered by DNA damage . These results demonstrate that fusion to the GFP color variants does not interfere with the functions of the P13010 in the Ku-dependent DSB repair . Therefore , these transformants might be useful not only in the analysis of P13010 behavior , but also in an analysis of the dynamics of the NHEJ repair process . 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 . 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 . The helix-loop-helix protein Id-1 and a retinoblastoma protein binding mutant of SV40 T antigen synergize to reactivate DNA synthesis in senescent human fibroblasts . Normal somatic cells of higher organisms do not divide indefinitely . After a finite number of divisions , normal cells irreversibly cease proliferation by a process termed replicative or cellular senescence . Replicative senescence is controlled by multiple , dominant-acting genes about which very little is known . The only genes known to reactivate DNA synthesis in senescent cells are viral oncogenes encoding proteins that bind and inactivate the p53 and retinoblastoma ( P06400 ) tumor suppressor proteins . SV40 T antigen is the best studied of these viral oncoproteins . T[ P04264 ] is a T antigen point mutant that selectively is defective in binding P06400 and the P06400 -related proteins P28749 and Q08999 . We show that T[ P04264 ] stimulated quiescent human fibroblasts to synthesize DNA nearly as well as wild-type T but was incapable of stimulating senescent cells . We tested several growth regulatory genes that are repressed in senescent cells for ability to restore activity to T[ P04264 ] . These included c-fos , c-jun , Id-1 , Id-2 , Q01094 , and cdc2 . Only the helix-loop-helix ( HLH ) protein , Id-1 , restored the ability of T[ P04264 ] to reactivate DNA synthesis in senescent cells . This activity of Id-1 was not shared by Id-2 , a related protein , and depended on an intact HLH domain . It did not appear that Id-1 interacted directly with P06400 or P28749 . Constitutive Id-1 expression failed to rescue proliferating cells from growth inhibition by P06400 , P28749 , or Q08999 , and failed to interact with P06400 in the yeast two hybrid system . Because Id proteins negatively regulate basic-HLH ( bHLH ) transcription factors , we suggest that senescent cells express one or more bHLH factor that cooperates with P06400 , or P06400 -related proteins , to suppress proliferation . DNA damage and S phase arrest induced by Ochratoxin A in human embryonic kidney cells ( P29320 293 ) . Ochratoxin A ( OTA ) is a ubiquitous mycotoxin with potential nephrotoxic , hepatotoxic and immunotoxic effects . The mechanisms underlying the nephrotoxicity of OTA remain obscure . To investigate DNA damage and the changes of the cell cycle distribution induced by OTA , human embryonic kidney cells ( P29320 293 cells ) were incubated with various concentrations of OTA for 24h in vitro . The results indicated that OTA treatment led to the production of reactive oxygen species ( ROS ) and to a decrease of the mitochondrial membrane potential ( ΔΨm ) . OTA-induced DNA damage in P29320 293 cells was evidenced by DNA comet tails formation and increased expression of γ- P16104 . In addition , OTA could induce cell cycle arrest at the S phase in P29320 293 cells . The expression of key cell cycle regulatory factors that were critical to the S phase , including cyclin A2 , cyclin E1 , and P24941 , were further detected . The expression of cyclin A2 , cyclin E1 , and P24941 were significantly decreased by OTA treatment at both the mRNA and protein levels . The apoptosis of P29320 293 cells after OTA treatment was observed using Hoechst 33342 staining . The results confirmed that OTA did induce apoptosis in P29320 293 cells . In conclusion , our results provided new insights into the molecular mechanisms by which OTA might promote nephrotoxicity . Effects of APRIL ( O75888 ) polymorphisms and splicing isoforms on the secretion of soluble APRIL . Functional APRIL ( O75888 ) is a secreted trimer generated by furin protease cleavage . We previously reported the association of APRIL haplotypes formed by two nonsynonymous polymorphisms , Gly67Arg and Asn96Ser , with systemic lupus erythematosus . Here , we tested the hypothesis that polymorphisms and/or alternative splicing may influence the generation of soluble APRIL ( sAPRIL ) . P29320 293T cells were transfected with plasmids containing one of the six combinations of splicing isoforms ( α or β ) and haplotypes ( susceptible , neutral , or protective ) . APRIL concentrations were quantitated in the cell lysates and supernatants using an enzyme-linked immunosorbent assay ( ELISA ) . The association between splicing efficiency and polymorphisms was analyzed using quantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The efficiency of cleavage by furin protease was analyzed using western blotting . Although both splicing isoforms were cleaved by furin protease , sAPRIL was not detected in the supernatant of the cells transfected with the β isoform , regardless of the haplotype . This suggested that , similarly to B-cell activating factor ( Q9Y275 ) , one of the major APRIL splicing isoforms may not be secreted as a functional molecule . Furthermore , the secretion of sAPRIL was decreased in the transfectants expressing the protective haplotype . An association between the polymorphisms and splicing efficiency or furin cleavage efficiency was not detected . In conclusion , these observations suggested that both alternative splicing and polymorphisms may affect the generation of functional sAPRIL . 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 . 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 ) . 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 . 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 . 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 . 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 . Developmental expression of CLC- P04264 in the postnatal rat kidney . CLC- P04264 , a kidney-specific chloride channel , has been demonstrated to be involved in the urine concentration mechanism . Here , we investigated the developmental expression of CLC- P04264 in the rat kidney . Using immunohistochemistry , we showed that CLC- P04264 was not present in the thin ascending limb of Henle 's loop during the early prenatal stages but was significantly expressed during the adult stage . CLC- P04264 started to appear at day 5 and its expression increased during further development . In developing rats this increase coincided with the increase in the urine-concentrating capacity as the animals matured . We also investigated the expressions of other channels and transporters , including Q13621 , P29972 , and P41181 . Q13621 was strongly expressed throughout the inner medulla in neonatal rat kidneys but was entirely undetectable at the adult stage . The decline in its expression took the form of a gradual recession from the inner medulla together with reciprocal increases in the expression of CLC- P04264 . P29972 was weakly expressed in the inner medulla during early development and showed a rapid increase in expression at a later stage . The collecting duct cells significantly expressed P41181 even at birth and maintained its expression throughout the development . These results suggest that CLC- P04264 expression is one of the major determinants of the urine-concentrating capacity of the developing rat kidney . 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 . Characterization of duck leucocytes by monoclonal antibodies . cDNAs encoding CD3epsilon , P01730 and the alpha-chain of CD8 of ducks were cloned . Monoclonal antibodies against Pekin duck P01730 and CD8alpha revealed that the antigens were expressed by the majority of thymocytes and by subpopulations of CD3+ cells in peripheral tissues . CD8alpha cell surface expression was also found on 90 % of bursal cells . The B cell specificity of a newly developed mab to the immunoglobulin light-chain ( L-chain ) was confirmed by double labelling studies with the chicken B-cell cytokine Q9Y275 . Using these tools and a mab reacting with the cytoplasmic domain of CD3epsilon , we demonstrated that the CD8alpha molecule is expressed to high levels in CD3-positive T cells and at lower levels in CD3-negative bursal and peripheral B cells . Mab 2-4 , which recognizes the chicken P10747 molecule , was found to react with P01730 -positive duck lymphocytes and with CD8-positive duck T cells but not with CD8-positive B cells . Mab P04264 , which recognizes a common antigen on chicken thrombocytes and monocytes/macrophages , was found to react with duck thrombocytes and macrophages . Thus , a range of mabs is now available which will allow to phenotype the major leucocyte populations in ducks , a surrogate infection model for hepatitis B virus .

MH_train_1143

MH_train_1143

MH_train_1143

interacts_with DB00945?

multiple_choice

DB00945 and age-related macular degeneration : positives versus negatives . The anti-inflammatory , analgesic , antipyretic and antithrombotic activities of aspirin confer its wide therapeutic application . The three former activities require higher doses of aspirin , whereas the latter can be achieved through a lower , thus safer dose of the drug . Low-dose , long-term aspirin is used as an antithrombotic therapy to prevent cardiovascular disease . Such therapy is used by millions of people worldwide , including those suffering from age-related macular degeneration ( AMD ) ; thus , questions have arisen as to whether such treatment has any impact on the development and course of AMD . This editorial addresses the important issue of possible beneficial and adverse effects of long-term , low-dose aspirin treatment of AMD patients . Special emphasis is given to the ability of aspirin to acetylate cyclooxygenases ( especially P35354 ) and thus to initiate a biochemical pathway leading to the generation of anti-inflammatory pro-resolving mediators synthesized from both ω-3 and ω-6 long-chain polyunsaturated fatty acids . Such mediators ( e.g. , resolvins , lipoxins ) may be of therapeutic value in retarding the development of dry form AMD . 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 . Gene-environment interactions in parkinsonism and Parkinson 's disease : the Geoparkinson study . OBJECTIVES : To investigate associations of Parkinson 's disease ( PD ) and parkinsonian syndromes with polymorphic genes that influence metabolism of either foreign chemical substances or dopamine and to seek evidence of gene-environment interaction effects that modify risk . METHODS : A case-control study of 959 prevalent cases of parkinsonism ( 767 with PD ) and 1989 controls across five European centres . Occupational hygienists estimated the average annual intensity of exposure to solvents , pesticides and metals , ( iron , copper , manganese ) , blind to disease status . P10635 , P27169 , P09488 , P30711 , P21266 , P09211 , P15559 , Q16678 , P21397 , P27338 , SOD 2 , P07099 , Q01959 , P14416 and NAT2 were genotyped . Results were analysed using multiple logistic regression adjusting for key confounders . RESULTS : There was a modest but significant association between P21397 polymorphism in males and disease risk ( G vs T , OR 1.30 , 95 % CI 1.02 to 1.66 , adjusted ) . The majority of gene-environment analyses did not show significant interaction effects . There were possible interaction effects between P09488 null genotype and solvent exposure ( which were stronger when limited to PD cases only ) . CONCLUSIONS : Many small studies have reported associations between genetic polymorphisms and PD . Fewer have examined gene-environment interactions . This large study was sufficiently powered to examine these aspects . P09488 null subjects heavily exposed to solvents appear to be at increased risk of PD . There was insufficient evidence that the other gene-environment combinations investigated modified disease risk , suggesting they contribute little to the burden of PD . Direct and irreversible inhibition of cyclooxygenase-1 by nitroaspirin ( DB05822 ) . Benzoic acid , 2-(acetyl-oxy)-3-[(nitrooxy)methyl]phenyl ester ( DB05822 ) , a new drug made by an aspirin molecule linked , through a spacer , to a nitric oxide ( NO ) -donating moiety , is now under clinical testing for the treatment of atherothrombotic conditions . DB00945 exerts its antithrombotic activity by irreversibly inactivating platelet cyclooxygenase ( P36551 ) -1 . DB05822 in vivo undergoes metabolism into deacetylated and/or denitrated metabolites , and it is not known whether DB05822 needs to liberate aspirin to inhibit P23219 , or whether it can block it as a whole molecule . The aim of our study was to evaluate the effects of DB05822 and its analog or metabolites on platelet P23219 and whole blood P35354 and on purified ovine P36551 ( oCOX ) -1 and oCOX-2 . In particular , we have compared the mechanism by which DB05822 inhibits purified oCOX enzymes with that of aspirin using a spectrophotometric assay . All the DB05822 derivatives containing acetylsalicylic acid inhibited the activity of oCOX-1 and oCOX-2 , whereas the deacetylated metabolites and the nitric oxide-donating moiety were inactive . Dialysis experiments showed that oCOX-1 inhibition by DB05822 , similar to aspirin , is irreversible . Reversible P36551 inhibitors ( indomethacin ) or salicylic acid incubated with the enzyme before DB05822 prevent the irreversible inhibition of oCOX-1 by DB05822 as well as by aspirin . In conclusion , our data show that DB05822 acts as a direct and irreversible inhibitor of P23219 and that the presence of a spacer and NO-donating moiety in the molecule slows the kinetics of P23219 inhibition by DB05822 , compared with aspirin . " DB00945 resistance " in ischemic stroke : insights using short thrombelastography . AIMS : DB00945 achieves its antithrombotic effect through inactivation of cyclo-oxygenase ( P36551 ) -1 , thereby preventing generation of thromboxane (TX)A2 from arachidonic acid ( AA ) . The reported prevalence of aspirin " resistance " varies significantly and is usually based on platelet function tests ( PFTs ) that use AA-induced platelet reactivity as a surrogate measure of the effect of aspirin , rather than specific assessment of its effect on its therapeutic target ( ie , P23219 inhibition ) . The reported rates are not only assay specific but also condition specific , with particularly high rates ( up to 70 % ) previously reported in the stroke population . We investigated whether pharmacological responses to aspirin can be reliably determined from a functional test of AA-induced whole-blood clotting . METHODS AND RESULTS : A prospective study included 35 patients admitted with ischemic stroke and commenced on 300 mg aspirin . AA-induced whole-blood clotting was measured using short thrombelastography , a previously extensively validated near-patient DB00522 . Serum TXB2 and inflammatory biomarkers were also measured . The prevalence of apparent aspirin resistance measured using AA was high ( range from 49 % to 67 % ) . However , serum [ TXB2 ] was consistently low , thereby confirming adequate inhibition of P23219 by aspirin . Mean inflammatory biomarker levels were elevated throughout . CONCLUSION : This study demonstrates that although P23219 activity is adequately and consistently suppressed by aspirin in stroke patients , this effect is not reliably indicated by whole-blood clotting in response to AA . These data help to explain why the reported prevalence of aspirin resistance in stroke from studies employing AA-induced platelet reactivity is high and cast doubt on the veracity of such reports . 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 . 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 . Prostaglandin-endoperoxide synthase genes P23219 and P35354 - novel modifiers of disease severity in cystic fibrosis patients . Cystic fibrosis ( CF ) is one of the most common autosomal recessive diseases among Caucasians caused by a mutation in the P13569 gene . However , the clinical outcome of CF pulmonary disease varies remarkably even in patients with the same P13569 genotype . This has led to a search for genetic modifiers located outside the P13569 gene . The aim of this study was to evaluate the effect of functional variants in prostaglandin-endoperoxide synthase genes ( P23219 and P35354 ) on the severity of lung disease in CF patients . To the best of our knowledge , it is the first time when analysis of P23219 and P35354 as potential CF modifiers is provided . The study included 94 CF patients homozygous for F508del mutation of P13569 . To compare their clinical condition , several parameters were recorded , e.g. a unique clinical score : disease severity status ( DSS ) . To analyse the effect of non- P13569 genetic polymorphisms on the clinical course of CF patients , the whole coding region of P23219 and selected P35354 polymorphisms were analysed . Statistical analysis of genotype-phenotype associations revealed a relationship between the heterozygosity status of identified polymorphisms and better lung function . These results mainly concern P35354 polymorphisms : -765G > C and 8473T > C . The P23219 and P35354 polymorphisms reducing P36551 protein levels had a positive effect on all analysed clinical parameters . This suggests an important role of these genes as protective modifiers of pulmonary disease in CF patients , due to inhibition of arachidonic acid conversion into prostaglandins , which probably reduces the inflammatory process . The P01584 -511 Polymorphism ( rs16944 AA Genotype ) Is Increased in DB00945 -Exacerbated Respiratory Disease in Mexican Population . DB00945 exacerbated respiratory disease ( AERD ) is characterized by chronic hyperplastic rhinosinusitis , nasal polyposis , asthma , and aspirin sensitivity . The mechanisms which produce these manifestations of intolerance are not fully defined , current research focuses on cyclooxygenase 1 ( P23219 ) inhibition , metabolism of arachidonic acid , and the P36551 pathway to the lipoxygenase ( LO ) route , inducing increased synthesis of leukotrienes ( LT ) . The biological plausibility of this model has led to the search for polymorphisms in genes responsible for proinflammatory cytokines synthesis , such as P01584 and P10145 . We performed a genetic association study between P10145 -251 ( rs4073 ) and P01584 -511 ( rs16944 ) polymorphisms in AERD , aspirin-tolerant asthma ( ATA ) , and healthy control subjects . Using allelic discrimination by real-time PCR , we found statistically nonsignificant associations between AERD , ATA , and healthy control subjects for the GG and GA genotypes of P01584 ( rs16944 ) . Interestingly , the AA genotype showed an increased frequency in the AERD patients versus the ATA group ( GF = 0.19 versus 0.07 , p = 0.018 , OR 2.98 , and 95 % CI 1.17-7.82 ) . This is the first observation that P01584 polymorphisms are involved in AERD . Thus , future studies must investigate whether interleukin-1β is released in the airways of AERD patients and whether it relates to genetic polymorphisms in the P01584 gene . Non steroidal anti-inflammatory drugs and P35354 inhibitors as anti-cancer therapeutics : hypes , hopes and reality . Non-steroidal anti-inflammatory drugs ( NSAIDs ) and specific inhibitors of cyclooxygenase ( P36551 ) -2 , are therapeutic groups widely used for the treatment of pain , inflammation and fever . There is growing experimental and clinical evidence indicating NSAIDs and P35354 inhibitors also have anti-cancer activity . Epidemiological studies have shown that regular use of DB00945 and other NSAIDs reduces the risk of developing cancer , in particular of the colon . Molecular pathology studies have revealed that P35354 is expressed by cancer cells and cells of the tumor stroma during tumor progression and in response to chemotherapy or radiotherapy . Experimental studies have demonstrated that P35354 over expression promotes tumorigenesis , and that NSAIDs and P35354 inhibitors suppress tumorigenesis and tumor progression . Clinical trials have shown that NSAIDs and P35354 inhibitors suppress colon polyp formation and malignant progression in patients with familial adenomatous polyposis ( FAP ) syndrome . Recent advances in the understanding of the cellular and molecular mechanisms of the anti-cancer effects of NSAIDs and P35354 inhibitors have demonstrated that these drugs target both tumor cells and the tumor vasculature . The therapeutic benefits of P35354 inhibitors in the treatment of human cancer in combination with chemotherapy or radiotherapy are currently being tested in clinical trials . In this article we will review recent advances in the understanding of the anti-tumor mechanisms of these drugs and discuss their potential application in clinical oncology . Elevated macrophage migration inhibitory factor and decreased transforming growth factor-beta levels in major depression -- no influence of celecoxib treatment . OBJECTIVES : The involvement of an immune process in the pathophysiology of major depression disorder ( MDD ) was substantiated by studies demonstrating elevated levels of proinflammatory cytokines and prostaglandin E(2) ( PGE(2) ) . P35354 ( P35354 ) inhibitors lead to a reduced production of PGE(2) and have been shown to improve depressive symptoms . We investigated the three immune parameters macrophage migration inhibitory factor ( MIF ) , transforming growth factor-β ( TGF-β ) and soluble P08571 ( sCD14 ) in a randomized , placebo-controlled trial of the P35354 inhibitor celecoxib as add-on therapy in patients with MDD treated with reboxetine . METHODS : Thirty-two patients with depression and 20 healthy controls participated in the study . The patients were treated with reboxetine and celecoxib or placebo . Immune parameters were measured from serum at baseline , after three and five weeks using ELISA . RESULTS : Celecoxib as add-on strategy resulted in a significant reduction of Hamilton Depression Scale scores compared to placebo . Depressed patients showed significantly elevated MIF ( p < 0.001 ) and reduced TGF-β ( p = 0.006 ) concentrations at baseline . There was no difference in sCD14-concentrations . There was no difference between the placebo and the celecoxib group and no change over time . LIMITATIONS : Limitations of the study are the relatively small sample size and lack of functional assessment of Q9Y251 axis in parallel . CONCLUSIONS : MIF is a promising new candidate in the neuro-immune interplay that may link depressive symptoms , altered immune state and Q9Y251 -axis dysregulation . Reduced levels of TGF-β replicate previous findings and support the importance of this regulatory cytokine in major depressive disorder . A novel family of hydroxamate-based acylating inhibitors of cyclooxygenase . DB00945 irreversibly inhibits cyclooxygenase ( P36551 ) by acetylating a serine residue in the active site . We synthesized a series of novel acylating agents based on our previously reported acetylating compound , O-acetylsalicylhydroxamic acid . One of these , triacetylsalicylhydroxamic acid ( TriAcSHA ) was more effective than aspirin and O-acetylsalicylhydroxamic acid in inactivating both P23219 and P35354 . Preincubation of P23219 with inhibitor for 5 min yielded IC(50) values of 18 microM for TriAcSHA and 60 microM for acetylsalicylic acid . Inhibition was time-dependent , with complete inhibition within 10 min at a concentration of 50 microM . As with aspirin , mutation of the serine 530 of P23219 to alanine abolished the activity of the TriAcSHA . Mutation of the alanine 119 to a glutamine markedly reduced the sensitivity to TriAcSHA , suggesting that this residue was necessary for the interaction with the enzyme . TriAcSHA was also more effective than aspirin as an inhibitor of platelet aggregation induced by arachidonic acid . The diacetylated phenylhydroxamates N-methyl-O,O-diacetylsalicylhydroxamic acid , N,O-diacetylbenzohydroxamic acid , and 2-methyl-O,N-diacetylbenzohydroxamic acid showed reduced or absent activity against P23219 . In addition , we synthesized a series of triacylsalicylhydroxamic acids with progressively longer acyl groups ( three to six carbons ) . All of the compounds inhibited P23219 and demonstrated progressively greater P23219 selectivity with increasing number of carbons . Hence , salicylhydroxamic acid provides a versatile backbone for the generation of a family of acylating inhibitors of cyclooxygenase . Q16552 promotes p38 MAPK-dependent endothelial activation enhancing neutrophil recruitment to sites of inflammation . Neutrophilic inflammation plays an important role in lung tissue destruction occurring in many chronic pulmonary diseases . Neutrophils can be recruited to sites of inflammation via the action of the cytokine Q16552 . In this study , we report that Q96F46 and Q8NAC3 mRNA expression is significantly increased in asthmatic bronchoscopic biopsies and that these receptors are not only expressed on epithelial and inflammatory cells but also on endothelial cells . Q16552 potently stimulates lung microvascular endothelial cells to produce chemoattractants ( P10145 and derivatives of the P09917 pathway ) that selectively drive neutrophil but not lymphocyte chemotaxis . Moreover , Q16552 promotes endothelial activation by inducing the expression of endothelial adhesion markers ( P16581 , P19320 , and P05362 ) in a p38 MAPK-dependent manner . This increased expression of adhesion molecules stimulates the trans-endothelial migration of neutrophils , as well as the transmigration of HT-29 colon carcinoma cells , suggesting a further role in promoting lung metastasis . Finally , Q16552 increased neutrophil adhesion to the endothelium in vivo as determined by intravital microscopy of mice cremaster muscle . Overall , our results demonstrate that Q16552 is a potent activator of the endothelium in vivo leading to neutrophil infiltration . Therefore , preventing neutrophil recruitment by blocking the action of Q16552 on endothelial cells may prove to be highly beneficial in diseases in which neutrophilic inflammation plays a key role . Hypothesis : decreased use of pediatric aspirin has contributed to the increasing prevalence of childhood asthma . BACKGROUND : The prevalence of asthma , atopic eczema , and allergic rhinitis has increased over the last three decades in Western countries . Speculation on the causes of this trend have focused on changes in environmental factors . We hypothesize that the decreased use of aspirin in favor of acetaminophen , due to the association of aspirin with Reye 's syndrome during febrile respiratory infections , may be contributing to these trends in the United States . DATA SOURCES : A detailed literature search was conducted utilizing Medline . Studies considered relevant and important involving both humans and animals in English language were used . HYPOTHESIS : In the United States , the documented prevalence of childhood asthma has increased since 1970 , but the rate of this increase accelerated upward beginning in the early 1980s when the use of pediatric aspirin decreased . During the resolution of common respiratory viral infections , prostaglandin E2 ( DB00917 ) is produced through the actions of cyclooxygenase-2 ( P35354 ) . DB00945 , but not acetaminophen , inhibits P35354 activity . As DB00917 promotes TH2 and inhibits THI type cytokine generation , we hypothesize that the decreased use of aspirin may be a factor in facilitating allergic sensitization and asthma by augmenting the relative Q8IXH7 /TH2 cytokine imbalance in genetically predisposed children . CONCLUSION : We have presented an hypothesis based upon epidemiologic trends , known biologic effects of cytokines and DB00917 on allergic sensitization , and a potentially relevant pharmacologic effect of aspirin to explain a component of the increasing prevalence of childhood asthma in the United States . We suggest this theory be examined further in animal models as well as in other countries where the prevalence of childhood asthma is increasing . Mechanistic and pharmacological issues of aspirin as an anticancer agent . Recent findings have shown that aspirin , taken for several years , reduces the long-term risk of some cancers , particularly colorectal cancer . The result that aspirin benefit is detectable at daily low-doses ( at least 75mg ) , the same used for the prevention of cardiovascular disease , positions the antiplatelet action of aspirin at the center of its antitumor efficacy . At low-doses given every 24 h , aspirin is acting by a complete and persistent inhibition of cyclooxygenase ( P36551 ) -1 in platelets ( in the pre-systemic circulation ) while causing a limited and rapidly reversible inhibitory effect on P35354 and/or P23219 expressed in nucleated cells . DB00945 has a short half-life in human circulation ( approximately 20 min ) ; nucleated cells have the ability to resynthesize the acetylated P36551 -isozymes within a few hours , while platelets do not . P36551 -independent mechanisms of aspirin , such as the inhibition of Wnt/ b-catenin and NF-kB signaling and the acetylation of extra- P36551 proteins , have been suggested to play a role in its chemo-preventive effects , but their relevance remains to be demonstrated in vivo at clinical doses . In conclusion , the results of clinical pharmacology and the analysis of randomized and epidemiological studies suggest that colorectal cancer and atherothrombosis share a common mechanism of disease , i.e. enhanced platelet activation in response to injury at distinct sites . Effects of combined octreotide and aspirin on the growth of gastric cancer . OBJECTIVE : To investigate the effects of the combination of octreotide and aspirin on the growth of gastric cancer . METHODS : Proliferation of gastric cancer cell lines treated with octreotide or aspirin was determined by (3)H-thymidine incorporation . After xenografts of human gastric cancer were implanted orthotopically in the stomach of nude mice , they were administered octreotide plus aspirin for 8 weeks . The mRNA of somatostatin receptor in the tissues of gastric carcinoma was detected by reverse transcription polymerase chain reaction ( RT-PCR ) . P35354 in gastric cancer tissues was measured by immunohistochemistry . RESULTS : Both octreotide and aspirin significantly reduced the (3)H-thymidine incorporation of gastric cancer cells . Xenografts in situ were found in all stomachs of nude mice except for two in the combination group . Either size or weight of tumors treated by octreotide , aspirin or in combination was significantly reduced as compared with that of controls . The inhibition rate for tumor was 60.6 % ( octreotide ) , 39.3 % ( aspirin ) , and 85.6 % ( in combination ) respectively . No severe side effects were observed in any treated groups . Somatostatin receptor-2 and -3 were expressed in the transplanted gastric adenocarcinomas . DB00945 could down-regulate the strong expression of cyclooxygenase-2 in the tissue of gastric adenocarcinomas of nude mice . CONCLUSION : A combination of octreotide and aspirin significantly inhibited proliferation of gastric cancer through mediation of somatostatin receptors and suppression of cyclooxygenase-2 . DB00945 , NSAIDs , and P35354 inhibitors in cardiovascular disease : possible interactions and implications for treatment of rheumatoid arthritis . DB00945 , nonsteroidal antiinflammatory drugs ( NSAIDs ) , and cyclooxygenase-2 ( P35354 ) inhibitors are widely used in patients with rheumatoid arthritis . DB00945 has the largest and most persuasive body of randomized trial evidence to support its use in secondary prevention for cardiovascular disease ( CVD ) and primary prevention for myocardial infarction . There is , however , a possible deleterious interaction between aspirin and NSAIDs on CVD that requires further research . DB00945 , NSAIDs , and to a lesser extent P35354 inhibitors are associated with increased gastrointestinal side effects and bleeding , alone and in combination . The more widespread and appropriate use of aspirin in patients with rheumatoid arthritis will avoid many premature deaths in secondary prevention for CVD and first myocardial infarctions in primary prevention . 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 . 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 . Association between type 2 diabetes genetic susceptibility loci and visceral and subcutaneous fat area as determined by computed tomography . Visceral fat accumulation has an important role in the development of several metabolic disorders , such as type 2 diabetes , dyslipidemia and hypertension . New genetic loci that contribute to the development of type 2 diabetes have been identified by genome-wide association studies . To examine the association of type 2 diabetes susceptibility loci and visceral fat accumulation , we genotyped 1279 Japanese subjects ( 556 men and 723 women ) , who underwent computed tomography for measurements of visceral fat area ( VFA ) and subcutaneous fat area ( SFA ) for the following single-nucleotide polymorphisms ( SNPs ) : Q04721 rs10923931 , Q6YHU6 rs7578597 , P37231 rs1801282 , Q9P2N4 rs4607103 , Q9Y6M1 rs1470579 , P15692 rs9472138 , Q86VZ6 rs864745 , CDKN2A/ P42772 rs564398 and rs10811661 , Q03014 rs1111875 and rs5015480 , Q9NQB0 rs7901695 , P51787 rs2237892 , Q14654 rs5215 and rs5219 , Q93063 rs1113132 , rs11037909 , and rs3740878 , P49286 rs10830963 , P81605 rs1153188 , P19075 / O75473 rs7961581 , and Q9C0B1 rs8050136 and rs9939609 . None of the above SNPs were significantly associated with VFA . The Q9C0B1 rs8050136 and rs9939609 risk alleles exhibited significant associations with body mass index ( BMI ; P=0.00088 and P=0.0010 , respectively ) and SFA ( P=0.00013 and P=0.00017 , respectively ) . No other SNPs were significantly associated with BMI or SFA . Our results suggest that two SNPs in the Q9C0B1 gene are associated with subcutaneous fat accumulation . The contributions of other SNPs are inconclusive because of a limitation of the sample power . Nonsteroidal anti-Inflammatory drugs and cardiovascular risk . Nonsteroidal anti-inflammatory drugs ( NSAID ) inhibit cyclooxygenase ( P36551 ) enzymes , which exist in at least two isoforms , P23219 and P35354 . DB00945 and older agents in this class are nonselective inhibitors of both P23219 and P35354 . Newer agents termed " coxibs " are selective inhibitors of P35354 . Among the NSAID , only aspirin has been proven to significantly reduce cardiovascular risk , primarily through inhibition of P23219 -mediated platelet aggregation . It has been suggested that other nonselective agents , especially naproxen , may provide some lesser degree of cardioprotection , but conclusive evidence is lacking . Conversely , there are concerns that the P35354 inhibitors may increase cardiovascular risk . However , mechanisms for this potentially adverse cardiovascular effect are unknown , and it is becoming increasingly clear that our understanding of the role of P35354 in cardiovascular function is incomplete . Some studies have demonstrated a potentially beneficial effect of P35354 on cardiovascular function that could be negated by P35354 inhibition , while other studies have reported improved endothelial function with P35354 inhibitors . Additionally , the impact of combined therapy with aspirin and other P36551 inhibitors is not yet clear . This article will review the studies that have examined these issues . Transcriptional regulation of prostaglandin-H synthase-2 gene in human trophoblasts . Abnormal PG production by placental PG-H synthase ( PGHS ) is associated with preeclampsia . There are two PGHS isozymes , and their regulation in trophoblasts is presently unknown . We hypothesized that the PGHS isozymes are differentially regulated in human trophoblasts . To test this hypothesis , we transfected primary trophoblasts and JEG3 cells with promoter constructs of either P23219 or P35354 genes . We found that in both cell systems , the basal activity of P35354 promoter was 10- to 30-fold higher than the activity of P23219 promoter . In response to either 12-0-tetradecanoylphorbol-13-acetate ( TPA ) or 8-bromo- DB02527 , we observed an increase in P35354 promoter activity but no change in activity of P23219 promoter . Similarly , both agents enhanced P35354 expression , as well as prostaglandin E2 production . The activity of P35354 promoter was potentiated by coexpression of protein kinase A and inhibited by coexpression of kinase A inhibitor . DB00945 attenuated the stimulatory effect of TPA on P35354 promoter . We conclude that both P23219 and P35354 promoters are active in trophoblasts . The activity of P35354 promoter is stimulated by either TPA or DB02527 , and the stimulatory effect of TPA is attenuated by aspirin . These pathways may play a role in modulation of prostanoid synthesis by trophoblasts . DB00945 resistance and genetic polymorphisms . Differences in genetic makeup or polymorphisms can affect individual drug response . Detecting genetic variation may help predict how a patient will respond to a drug and could be used as a tool to select optimal therapy , tailor dosage regimens , and improve clinical outcomes . The data are replete relative to the therapeutic efficacy of aspirin ( ASA ) for the prevention of ischemic events . However , there is a paucity of published data on the relationship between polymorphisms and the clinical effects on ASA . Prothrombotic genetic variations that may contribute to ASA resistance , and increased risk of cardiovascular events may involve : ( 1 ) a polymorphism on the cyclooxygenase-1 ( P23219 ) gene affecting Ser529 ; ( 2 ) overexpression of P35354 mRNA on platelets and endothelial cells ; ( 3 ) polymorphism Q9UKI9 /A2 of the gene encoding glycoprotein IIIa ( P05106 ) ; and ( 4 ) the homozygous 807T ( 873A ) polymorphism allied with increased density of platelet GP Ia/IIa collagen-receptor gene . Because of the possible increased risk of ischemic vascular events , carriers of these genetic polymorphisms may be resistant to the antithrombotic effects of ASA and should be considered for additional or alternative treatment . 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 . Role of prostacyclin in the cardiovascular response to thromboxane A2 . Thromboxane ( Tx ) A2 is a vasoconstrictor and platelet agonist . DB00945 affords cardioprotection through inhibition of TxA2 formation by platelet cyclooxygenase ( P23219 ) . DB01240 ( DB01240 ) is a vasodilator that inhibits platelet function . Here we show that injury-induced vascular proliferation and platelet activation are enhanced in mice that are genetically deficient in the P43119 ( IP ) but are depressed in mice genetically deficient in the TxA2 receptor ( TP ) or treated with a TP antagonist . The augmented response to vascular injury was abolished in mice deficient in both receptors . Thus , DB01240 modulates platelet-vascular interactions in vivo and specifically limits the response to TxA2 . This interplay may help explain the adverse cardiovascular effects associated with selective P35354 inhibitors , which , unlike aspirin and nonsteroidal anti-inflammatory drugs ( NSAIDs ) , inhibit DB01240 but not TxA2 . 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 . [ The concept of aspirin " resistance " : mechanisms and clinical relevance ] . DB00945 , a 110-year-old molecule , is a cornerstone in the treatment of atherothrombotic patients . The concept of aspirin " resistance " emerged approximately 15 years ago and is of growing interest . DB00945 resistance , defined as a lack of inhibition of cyclo-oxygenase-1 ( P23219 ) , is a rare phenomenon and its clinical relevance can hardly be studied . On the contrary , residual platelet hyperactivity is more common and affects 20 to 30 % of aspirin-treated patients . This latter phenomenon corresponds to sustained platelet reactivity despite a proper inhibition of P23219 by aspirin . Several meta-analyses suggest that residual platelet hyperactivity could be a risk factor for the recurrence of ischemic events in aspirin-treated patients . Causes of biological non-responsiveness to aspirin are discussed , including the role of compliance , drug-drug interactions , genetic polymorphisms and diabetes mellitus . Ongoing studies are designed to find out the mechanisms of residual platelet hyperactivity , determine its potential clinical relevance and delineate the more appropriate assays in order to identify patients who may benefit of a tailored antiplatelet therapy . Dopamine receptors and psychiatric drug treatment . The established antipsychotic drugs act mainly by antagonizing dopamine mediated synaptic transmission in the brain . Increase in the rate of production of dopamine metabolites as well as the firing rate of dopamine-containing neurons can be interpreted as compensatory responses to an interruption of synaptic transmission at dopamine nerve terminals . The demonstration of involvement of limbic and cortical mechanisms in the antipsychotic activity of neuroleptic drugs is far more difficult than the involvement of nigro-striatal and tubero-infundibular mechanisms in the neurological and neuroendocrine effects of these drugs . Application of radioreceptor techniques to dopamine research has supported the findings obtained by other neuropsychopharmacological research techniques , providing more direct evidence of dopamine receptor blockade by neuroleptic drugs . Further research is needed especially in studying the nature of the time-dependent adaptive changes at the receptor sites as well as the differences between the different dopamine projections and neural systems in the brain . The different subtypes of dopamine receptors in the brain , currently called D1 and D2 dopamine receptors , seem to be parallel , although in many respects independently-acting regulatory systems . P14416 -selective antagonists such as sulpiride seem to cause selective D2 receptor up-regulation . P01236 secretion seems to be regulated by D2 dopamine receptors . The exact physiological role of D1 dopamine receptors as well as the clinical consequences of selective D1 antagonism is not known . DB00391 and clozapine are examples of atypical neuroleptic compounds that have quite different profile of action , the former having strong and selective antidopaminergic action , the latter combining a number of non- dopaminergic mechanisms with rather slight effects on dopamine receptors. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . miRNA signature and Dicer requirement during human endometrial stromal decidualization in vitro . Decidualization is a morphological and biochemical transformation of endometrial stromal fibroblast into differentiated decidual cells , which is critical for embryo implantation and pregnancy establishment . The complex regulatory networks have been elucidated at both the transcriptome and the proteome levels , however very little is known about the post-transcriptional regulation of this process . miRNAs regulate multiple physiological pathways and their de-regulation is associated with human disorders including gynaecological conditions such as endometriosis and preeclampsia . In this study we profile the miRNAs expression throughout human endometrial stromal ( hESCs ) decidualization and analyze the requirement of the miRNA biogenesis enzyme Dicer during this process . A total of 26 miRNAs were upregulated and 17 miRNAs downregulated in decidualized hESCs compared to non-decidualized hESCs . Three miRNAs families , miR-181 , miR-183 and miR-200 , are down-regulated during the decidualization process . Using miRNAs target prediction algorithms we have identified the potential targets and pathways regulated by these miRNAs . The knockdown of Dicer has a minor effect on hESCs during in vitro decidualization . We have analyzed a battery of decidualization markers such as cell morphology , P01236 , P08833 , P55773 and P35625 secretion as well as P31260 , P35354 , SP1 , C/EBPß and Q12778 expression in decidualized hESCs with decreased Dicer function . We found decreased levels of P31260 and altered intracellular organization of actin filaments in Dicer knockdown decidualized hESCs compared to control . Our results provide the miRNA signature of hESC during the decidualization process in vitro . We also provide the first functional characterization of Dicer during human endometrial decidualization although surprisingly we found that Dicer plays a minor role regulating this process suggesting that alternative biogenesis miRNAs pathways must be involved in human endometrial decidualization . Reed-Sternberg cell-derived lymphotoxin-α activates endothelial cells to enhance T-cell recruitment in classical Hodgkin lymphoma . It is known that cells within the inflammatory background in classical Hodgkin lymphoma ( cHL ) provide signals essential for the continual survival of the neoplastic Hodgkin and Reed-Sternberg ( HRS ) cells . However , the mechanisms underlying the recruitment of this inflammatory infiltrate into the involved lymph nodes are less well understood . In this study , we show in vitro that HRS cells secrete lymphotoxin-α ( LTα ) which acts on endothelial cells to upregulate the expression of adhesion molecules that are important for T cell recruitment . LTα also enhances the expression of hyaluronan which preferentially contributes to the recruitment of P01730 (+) CD45RA(+) naïve T cells under in vitro defined flow conditions . Enhanced expression of LTα in HRS cells and tissue stroma ; and hyaluronan on endothelial cells are readily detected in involved lymph nodes from cHL patients . Our study also shows that although NF-κB and AP-1 are involved , the cyclooxygenase ( P36551 ) pathway is the dominant regulator of LTα production in HRS cells . Using pharmacological inhibitors , our data suggest that activity of P23219 , but not of P35354 , directly regulates the expression of nuclear c-Fos in HRS cells . Our findings suggest that HRS cell-derived LTα is an important mediator that contributes to T cell recruitment into lesional lymph nodes in cHL . Differing profiles of prostaglandin formation inhibition between selective prostaglandin H synthase-2 inhibitors and conventional NSAIDs in inflammatory and non-inflammatory sites of the rat . The present study examined the inhibitory profiles of NS-398 and nimesulide against prostaglandin ( PG ) formation in inflammatory and non-inflammatory sites , and compared them with those of aspirin and indomethacin . In vitro , indomethacin inhibited PGH synthase ( PGHS ) -1 and P35354 almost equally , while NS-398 and nimesulide inhibited only P35354 . NS-398 ( 1 , 10 mg/kg ) and nimesulide ( 3 mg/kg ) slowed the rate of plasma exudation and thus the exudate accumulation in rat carrageenin-induced pleurisy . DB00945 ( 30 , 100 mg/kg ) and indomethacin ( 10 mg/kg ) also reduced this rate . NS-398 and nimesulide reduced the DB00917 more potently than TXB2 and 6-keto-PGF1 alpha in the exudate . However , aspirin and indomethacin did not exhibit this selectivity . The levels of DB00917 correlated significantly with the plasma exudation rate . Moreover , nimesulide ( 3 mg/kg ) did not affect DB00917 formation in rat stomachs injected with 1 M NaCl solution , while indomethacin ( 10 mg/kg ) reduced it . Thus , NS-398 and nimesulide exhibit different inhibitory profiles from aspirin and indomethacin against PG formation . These results suggest that DB00917 may be produced by P35354 in the inflammatory site , and may play a more prominent role than DB01240 in plasma exudation . DB00945 induction of apoptosis in esophageal cancer : a potential for chemoprevention . The potential use of non-steroidal anti-inflammatory drugs ( NSAIDs ) in the prevention of gastrointestinal cancers has been highlighted recently . However , it is not known whether NSAIDs could also be useful for preventing esophageal cancer , although regular users of these drugs appear to have a decreased incidence of esophageal cancer . Therefore , we examined the effect of aspirin on growth and apoptosis in 10 esophageal cancer cell lines as well as the expression and modulation of its target enzymes , cyclooxygenases ( COXs ) , and their product prostaglandin E2 . Growth inhibition of these cells by aspirin was dose- and time-dependent and associated with the induction of apoptosis . P23219 and P35354 were expressed in 7 of the 10 cell lines . Bile acids could induce P35354 expression in six of eight cell lines tested , which was correlated with prostaglandin E2 production , and aspirin could inhibit P35354 enzymatic activity even after bile acid stimulation but was unable to change the P35354 protein level in these cell lines . Down-regulation of bcl-2 by aspirin was found in the two cell lines tested . These results suggest that induction of apoptosis by aspirin may be a mechanism by which it can intervene in esophageal carcinogenesis and may be indicative of the potential of NSAIDs as chemopreventive agents in esophageal cancer . DB00945 -exacerbated asthma . : This review focuses on aspirin-exacerbated asthma ( AEA ) . The review includes historical perspective of aspirin , prevalence , pathogenesis , clinical features and treatment of AEA . The pathogenesis of AEA involves the cyclooxygenase and lipooxygenase pathway . DB00945 affects both of these pathways by inhibiting the enzyme cycooxygenase-1 ( P23219 ) . Inhibition of P23219 leads to a decrease in prostaglandin E2 ( DB00917 ) . The decrease in DB00917 results in an increase in cysteinyl leukotrienes by the lipooxygenase pathway involving the enzyme 5-lipooxygenase ( P09917 ) . Leukotriene C4 ( LTC4 ) synthase is the enzyme responsible for the production of leukotriene C4 , the chief cysteinyl leukotriene responsible for AEA . There have been familial occurences of AEA . An allele of the Q16873 gene in AEA is known as allele C . Allele C has a higher frequency in AEA . Clinical presentation includes a history of asthma after ingestion of aspirin , nasal congestion , watery rhinorrhea and nasal polyposis . Treatment includes leukotriene receptor antagonists , leukotriene inhibitors , aspirin desinsitaztion and surgery . AEA is the most well-defined phenotype of asthma . Although AEA affects adults and children with physician-diagnosed asthma , in some cases there is no history of asthma and AEA often goes unrecognized and underdiagnosed . Compound FLZ inhibits lipopolysaccharide-induced inflammatory effects via down-regulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways in RAW264.7 macrophages . AIM : The aim of this study was to investigate the effect of the squamosamide derivative FLZ ( N-2-(4-hydroxy-phenyl)-ethyl-2-(2,5-dimethoxy-phenyl)-3-(3-methoxy-4-hydroxy-phenyl)-acrylamide ) on lipopolysaccharide ( LPS ) -induced inflammatory mediator production and the underlying mechanism in RAW264.7 macrophages . METHODS : RAW264.7 cells were preincubated with non-toxic concentrations of compound FLZ ( 1 , 5 , and 10 micromol/L ) for 30 min and then stimulated with 10 microg/L LPS . The production of nitric oxide ( NO ) , the expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase 2 ( P35354 ) , and the activation of nuclear factor kappa-B ( NF-kappaB ) and mitogen-activated protein kinase ( MAPK ) pathways were examined . RESULTS : FLZ significantly inhibited the LPS-induced production of NO , as well as the expression of P35228 and P35354 at both the RNA and the protein levels in RAW264.7 cells . The LPS-induced increase in the DNA binding activity of NF-kappaB and activator protein 1 ( AP-1 ) , the nuclear translocation of NF-kappaB p65 , the degradation of the inhibitory kappaBalpha protein ( P25963 ) and the phosphorylation of P25963 , O15111 ( IKK ) alpha/beta , c-Jun NH(2)-terminal kinase ( JNK ) and p38 MAPKs were all suppressed by FLZ . However , the phosphorylation of extracellular signal-regulated kinase ( P29323 ) was not affected . Further study revealed that FLZ inhibited the phosphorylation of transforming growth factor-beta ( TGF-beta ) -activated kinase 1 ( TAK1 ) , which is an upstream signaling molecule required for IKKalpha/beta , JNK and p38 activation . CONCLUSION : FLZ inhibited the LPS-induced production of inflammatory mediators at least partly through the downregulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways . [ DB00945 and hemostasis ] . DB00945 is one hundred years old , though its use has clearly evolved during the last 25 years . Identifying its action mechanism has allowed us to better understand the antithrombotic impact . Prostaglandin H synthetase ( PGHS ) is a bifunctional enzyme with cyclooxygenase and peroxydase activities . There are two isoforms : constitutive P23219 and inducible P35354 . DB00945 irreversive acetylates the platelet cyclooxygenase involved in the formation of thromboxane A2 , a powerful proaggregating agent and vasoconstrictor . More than 95 % of inhibition of this synthesis takes place in two to three days using very weak doses of aspirin , on the order of 30 to 50 mg per day . Under some circumstances , this inhibition requires higher dosages . Certain clinical and biological circumstances could lead to a resistance to aspirin , making a readjustment of doses and sometimes complementary explorations necessary . The ISIS 2 study showed in an apparently irrefutable way the entry of aspirin into the antithrombotics arsenal , with a significant risk reduction of vascular death and recurrence of infarctus . Numerous studies have confirmed this efficacy . Consensus studies are based on information showing total coherence between the dose necessary to acetylate the enzyme to inhibit thromboxane A2 platelet production and the clinical antithrombotic effect . DB00945 seems to have a secure place , and it begins the third millennium in relative peace with new extra-platelet potentialities outside the framework of hemostasis and thrombosis . Neuroprotective and anti-inflammatory activities of atorvastatin in a rat chronic constriction injury model . DB01076 is an P04035 inhibitor used to treat hypercholesterolemic conditions associated with hypertension . This study aims to investigate the anti-inflammatory and neuroprotective effects of atorvastatin on peripheral neuropathic pain . Peripheral neuropathic pain was induced by chronic constriction injury ( CCI ) in Sprague-Dawley rats . Rats were divided into 3 groups including sham-operated , CCI , and atorvastatin-treated . DB01076 ( 10 mg/kg ) or phosphate-buffered saline was orally administered for 2 weeks . All animals were assessed by neurobehavioral tests before surgery and at days 3 , 7 , 14 after surgery . Inflammatory and neuroprotective factors were evaluated by Western blot analysis . P29474 , P35354 and P35228 in the sciatic nerve were also studied using immunohistochemistry . DB01076 attenuated CCI-induced nociceptive sensitization and thermal hyperalgesia in a time-dependent manner . DB01076 improved CCI-induced neurobehavioral/inflammatory activity by inhibition of TGF-beta , pIkB/IkB , NFkB , P35354 , P35228 , EP1 and EP4 in the sciatic nerve . DB01076 was also found to increase neuroprotection factors pAkt/Akt , P29474 and P15692 . Taken together , these data indicate that atorvastatin could protect the sciatic nerve against CCI-induced neuroinflammation and nociception . 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 . 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 . Luteolin supplementation adjacent to aspirin treatment reduced dimethylhydrazine-induced experimental colon carcinogenesis in rats . Previous studies have shown that aspirin is used in colon cancer treatment . However , long-term of DB00945 usage is limited to gastric and renal toxicity . Luteolin ( LUT ) has cancer prevention and anti-inflammatory effects . The present study was designed to investigate the effect of LUT supplementation and DB00945 treatment in dimethylhydrazine ( Q03001 ) -induced carcinogenesis in rats . Q03001 ( 20 mg/kg BW/week ) treated rats received gavages with DB00945 ( 50 mg/kg BW/week ) and LUT ( 0.2 mg/kg BW/day ) for 15 weeks . Q03001 injections induce colon polyps and renal bleeding , significantly increasing carcinoembryonic antigen ( P06731 ) , cyclooxygenase-2 ( P35354 ) , oxidative stress , and kidney function tests and reducing antioxidant markers . Either DB00945 or LUT gavages alone or combined produce a significant decrease in colon polyp number and size , significantly decreasing P06731 , P35354 , and oxidative stress and increasing antioxidant markers . In conclusion , the supplementations of LUT adjacent to DB00945 in the treatment of Q03001 -induced carcinogenesis in rats reflect a better effect than the use of DB00945 alone . DB00945 causes rapid up-regulation of cyclo-oxygenase-2 expression in the stomach of rats . BACKGROUND : Cyclo-oxygenase-1 ( P23219 ) is believed to produce prostaglandins vital to mucosal defence , whereas cyclo-oxygenase-2 ( P35354 ) is induced at sites of inflammation . Little is known about the regulation of P35354 in the stomach , particularly during the period following mucosal injury . In this study , we examined P23219 and P35354 expression shortly after administration of NSAIDs or ethanol . METHODS : Fasted rats were given aspirin , salicylate , indomethacin or ethanol ( 20 % or 40 % ) orally . Three hours later the stomach was excised , the severity of damage scored and samples taken for RT-PCR of P23219 and P35354 mRNA and immunohistochemistry . DB00435 synthase mRNA ( P35228 and P29474 ) and activity were also measured . RESULTS : DB00945 , indomethacin and the higher concentration of ethanol produced widespread mucosal damage , whereas salicylate and 20 % ethanol caused only superficial epithelial damage . DB00945 caused a significant increase in P35354 mRNA expression and a marked increase in P35354 immunoreactivity , particularly in the superficial mucosa . Expression of P23219 ( mRNA and protein ) was unaffected by aspirin , as were NOS mRNA expression and enzyme activity . Pre-treatment with prostaglandin E2 prevented the induction of P35354 by aspirin . Salicylate and indomethacin caused modest increases in P35354 immunoreactivity but no change in P35354 mRNA . Neither concentration of ethanol affected P35354 mRNA or protein expression , suggesting that this was a specific response to the aspirin , rather than to injury . CONCLUSIONS : These results demonstrate a rapid up-regulation of P35354 expression in response to aspirin , possibly representing a compensatory response to inhibition of gastric prostaglandin synthesis . The prognostic value of metalloproteinases and angiogenic factors in ovarian carcinoma . The objective of this study was to analyze the correlation between matrix metalloproteinases ( MMPs ) and angiogenic genes and survival in advanced-stage ovarian carcinomas . Primary and metastatic ovarian carcinomas from patients diagnosed with FIGO stage III-IV disease and followed up to 20 years were studied using mRNA in situ hybridization ( ISH ) . Expression of P08253 , P14780 , membrane-type 1-MMP ( P50281 ) , the MMP inhibitor P16035 , vascular endothelial growth factor ( P15692 ) , interleukin-8 ( P10145 ) and basic fibroblast growth factor ( P09038 ) was studied . P08253 , P14780 and P16035 mRNA was detected in both tumor and stromal cells , while P50281 was largely confined to tumor cells . In univariate analysis of primary tumors , P16035 and P14780 mRNA expression correlated with poor outcome . In metastatic lesions , mRNA expression of P16035 , P08253 , and P50281 correlated with poor survival . In a multivariate analysis of primary tumors , P16035 expression in stromal cells ( P=0.006 ) and P14780 expression in tumor cells ( P=0.011 ) retained their predictive value . Intense expression of P09038 mRNA and weak expression of P10145 mRNA was detected in both stromal and tumor cells in most cases , while P15692 mRNA expression was limited to a few cases . Angiogenic mRNA expression showed no correlation with disease outcome in survival analysis ( P > 0.05 ) . We conclude that P09038 is the major angiogenic factor expressed in ovarian carcinoma at the mRNA level . P08253 , P14780 , P50281 and P16035 are valid markers of poor survival in advanced-stage ovarian carcinoma . 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 . Determination of expression of cyclooxygenase-1 and -2 isozymes in canine tissues and their differential sensitivity to nonsteroidal anti-inflammatory drugs . OBJECTIVE : To evaluate cyclooxygenase isozyme distribution in tissues from dogs and determine the differential sensitivity of canine cyclooxygenase ( P36551 ) -1 and -2 isozymes to nonsteroidal anti-inflammatory drugs ( NSAIDs ) . SAMPLE POPULATION : Canine tissue samples ( stomach , duodenum , ileum , jejunum , colon , spleen , cerebral cortex , lung , ovary , kidney , and liver ) were obtained from 2 dogs for northern and western blot analyses , and blood for whole blood P36551 assays was obtained from 15 dogs . PROCEDURE : 11 NSAIDs were evaluated to determine their P35354 selectivity in whole blood assays . The concentrations of the drug needed to inhibit 50 % of enzyme activity ( IC50 ) were then calculated for comparison . Expression and tissue distribution of P36551 isozymes were determined by northern and western blot analysis . RESULTS : DB00945 , diclofenac , indomethacin , ketoprofen , meclofenamic acid , and piroxicam had little selectivity toward P36551 isozymes , whereas NS398 , carprofen , tolfenamic acid , nimesulide , and etodolac had more than 5 times greater preference for inhibiting P35354 than P23219 . All canine tissues examined , including those from the gastrointestinal tract , coexpressed P23219 and -2 mRNA , although protein expression was observed only for P23219 . CONCLUSIONS AND CLINICAL RELEVANCE : Canine P35354 was selectively inhibited by etodolac , nimesulide , and NS398 ; tolfenamic acid and carprofen also appeared to be preferential P35354 inhibitors in dogs . The roles of P23219 as a constitutive housekeeping enzyme and P35354 as a proinflammatory inducible enzyme ( as determined in humans ) appear to apply to dogs ; therefore , P35354 -selective inhibitors should prove useful in reducing the adverse effects associated with nonselective NSAIDs . P35354 -dependent and -independent biosynthesis of dihydroxy-arachidonic acids in activated human leukocytes . Biosynthesis of 5,15-dihydroxyeicosatetraenoic acid ( 5,15-diHETE ) in leukocytes involves consecutive oxygenation of arachidonic acid by P09917 ( P28300 ) and P16050 in either order . Here , we analyzed the contribution of cyclooxygenase ( P36551 ) -2 to the biosynthesis of 5,15-diHETE and 5,11-diHETE in isolated human leukocytes activated with lipopolysaccharide and calcium ionophore A23187 . Transformation of arachidonic acid was initiated by 5- P28300 providing 5S-HETE as a substrate for P35354 forming 5S,15S-diHETE , 5S,15R-diHETE , and 5S,11R-diHETE as shown by LC/MS and chiral phase HPLC analyses . The levels of 5,15-diHETE were 0.45 ± 0.2 ng/10⁶ cells ( mean ± SEM , n = 6 ) , reaching about half the level of LTB₄ ( 1.3 ± 0.5 ng/10⁶ cells , n = 6 ) . The P35354 specific inhibitor NS-398 reduced the levels of 5,15-diHETE to below 0.02 ng/10⁶ cells in four of six samples . Similar reduction was achieved by MK-886 , an inhibitor of 5- P28300 activating protein but the above differences were not statistically significant . DB00945 treatment of the activated cells allowed formation of 5,15-diHETE ( 0.1 ± 0.05 ng/10⁶ cells , n = 6 ) but , as expected , abolished formation of 5,11-diHETE . The mixture of activated cells also produced 5S,12S-diHETE with the unusual 6E,8Z,10E double bond configuration , implicating biosynthesis by 5- P28300 and P16050 activity rather than by hydrolysis of the leukotriene A₄-epoxide . Exogenous octadeuterated 5S-HETE and 15S-HETE were converted to 5,15-diHETE , implicating that multiple oxygenation pathways of arachidonic acid occur in activated leukocytes . The contribution of P35354 to the biosynthesis of dihydroxylated derivatives of arachidonic acid provides evidence for functional coupling with 5- P28300 in activated human leukocytes . 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 . DB00945 -triggered lipoxin in patients treated with aspirin and selective vs. nonselective P35354 inhibitors . AIMS : Cyclooxygenase ( P36551 ) -2 inhibition has been reported to suppress the biosynthesis of the gastroprotective lipoxygenase metabolite 15(R)-epi-lipoxin A(4) , also termed ' aspirin-triggered lipoxin ' ( ATL ) . We tested the hypothesis that the co-administration of aspirin with either the selective P35354 inhibitor celecoxib or the nonselective P36551 inhibitor ibuprofen reduces ATL biosynthesis . METHODS : We measured the urinary excretion of ATL in 24 patients with both ischaemic heart disease and osteoarthritis , chronically treated with aspirin and co-administered celecoxib 200 mg b.i.d. , ibuprofen 600 mg t.i.d. , or placebo for 7 days . RESULTS : Baseline ATL was comparable in the three groups . On days 1 and 7 , 4 h after co-administration of celecoxib or ibuprofen , ATL levels did not show significant variations ( day 1 : 0.24 +/- 0.33 , 0.26 +/- 0.21 and 0.37 +/- 0.22 ng mg(-1) creatinine , respectively ; day 7 : 0.21 +/- 0.13 , 0.35 +/- 0.15 and 0.23 +/- 0.18 ng mg(-1) creatinine , respectively ) . CONCLUSIONS : Neither selective nor nonselective P35354 inhibition appreciably interferes with ATL biosynthesis , suggesting that this mediator is not involved in exacerbating gastrotoxicity by the association of aspirin with P35354 inhibitors . DB00945 twice a day keeps new P23219 at bay . 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 . Safe full-dose one-step nabumetone challenge in patients with nonsteroidal anti-inflammatory drug hypersensitivity . DB00945 and all nonsteroidal anti-inflammatory drugs ( NSAIDs ) are a chemically heterogeneous group of compounds that share the ability to inhibit the enzyme cyclooxygenase ( P36551 ) . This inhibitory effect , especially of P23219 , is suggested as the mechanism underlying NSAID-induced hypersensitivity reactions . In this study , we evaluated the safety and convenience of a single full-dose challenge with nabumetone , a selective P35354 inhibitor , in patients with hypersensitivity to nonselective NSAIDs ( ns-NSAIDs ) . Twenty-four subjects with a history of hypersensitivity reactions to at least two different ns-NSAIDs on two different occasions were enrolled in the study . The patients were otherwise healthy and did not suffer from NSAID- or aspirin-induced asthma or urticaria . All subjects were orally challenged by a single full dose ( 1000 mg ) of nabumetone , monitored closely in the hospital for the next 4 hours and contacted by telephone the next morning and 3-12 months afterward . Twenty-two patients tolerated nabumetone without any reaction during and after the challenge . One patient had a single urticarial lesion and one patient reported mild pruritus without objective signs , both of which resolved spontaneously . Thirteen patients , including the patient who responded with pruritus to the challenge , used nabumetone on several occasions during the follow-up period without any adverse reaction . Our study shows that in patients with a history of aspirin- and ns-NSAID-induced hypersensitivity reaction , a rapid one-step challenge with nabumetone was well tolerated . These initial data support the possibility that a single full dose of nabumetone can be tried as a safe alternative in most patients with a hypersensitivity reaction to ns-NSAIDs . The aspirin metabolite , salicylate , inhibits 7,12-dimethylbenz[a]anthracene-DNA adduct formation in breast cancer cells . There is evidence that aspirin and other non-steroidal anti-inflammatory drugs may be protective agents against cancer in the gastrointestinal tract . These effects are particularly well documented for the colon and rectum . Some epidemiological and experimental studies have suggested that aspirin could also be a chemopreventive agent against breast cancer . We investigated the effects of the aspirin metabolite , salicylate ( SA ) , on 7,12-dimethylbenz[a]anthracene ( DMBA ) -DNA adduct formation as well as on the expression of the enzymes involved in the carcinogen bioactivation pathway , in particular cytochrome P450 1A ( CYP1A ) and cyclooxygenases ( P23219 and P35354 ) . The effects of the test drug were examined in both the human mammary carcinoma cell line , MCF-7 , and mammary cells derived from DMBA-induced rat mammary tumours ( RMTCs ) . In this study , we also reported the effects of SA on cell growth and viability in breast cancer cells ( BCCs ) . The results demonstrated that DMBA-DNA adduct formation in both cancer cell lines was inhibited by SA at concentrations of > or = 2.5 mM . CYP1A was undetectable in RMTCs while CYP1A induction by beta-naphthoflavone in MCF-7 cells was significantly inhibited by SA in a concentration-dependent manner . DB00945 did not affect P23219 expression in either of the BCCs . P35354 was not detected in MCF-7 cells , but its expression in RMTCs was inhibited by SA treatment , which also significantly reduced BCC growth , but failed to cause cell death by necrosis or apoptosis . These data suggest that inhibition of DMBA-DNA adduct formation may contribute to aspirin breast cancer chemopreventive action and indicate that this drug can act in the first stage of carcinogenesis . [ 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 . Mechanism of chronic urticaria exacerbation by aspirin . In some patients with chronic idiopathic urticaria ( CIU ) , aspirin and other nonsteroidal anti-inflammatory drugs ( NSAIDs ) that inhibit cyclooxygenase 1 ( P23219 ) precipitate wheals and swelling . There is no in vitro diagnostic , and diagnosis can be established only by provocation challenges with aspirin or other NSAIDs . Skin reactions triggered by aspirin are associated with the inhibition of cyclooxygenase , specifically P23219 , but not P35354 , and are characterized by overproduction of cysteinyl leukotrienes ( cys-LTs ) . DB00945 and other NSAIDs should be avoided , but highly specific P35354 inhibitors , known as coxibs , are well tolerated and can probably be safely used . Evidence has been accumulated that these reactions are due to the interference of aspirin-like drugs with arachidonic-acid metabolism . In this article , we discuss the mechanism of these reactions , and the characteristic course of aspirin-induced urticaria and its management . DB00945 attenuates cytomegalovirus infectivity and gene expression mediated by cyclooxygenase-2 in coronary artery smooth muscle cells . Human cytomegalovirus ( CMV ) infection of smooth muscle cells generates reactive oxygen species ( ROS ) and thereby activates nuclear factor kappaB ( NFkappaB ) , which causes expression of viral and cellular genes involved in immune and inflammatory responses . These changes could account for the mounting evidence suggesting that CMV may contribute causally to restenosis and atherosclerosis . We found that CMV induces ROS , at least partly , through a cyclooxygenase-2 ( P35354 ) -dependent pathway . Moreover , the viral immediate-early ( IE ) gene products , IE72 and IE84 , have the capacity to transactivate the P35354 promoter . DB00945 and indomethacin , both cyclooxygenase inhibitors as well as direct ROS scavengers , reduce CMV-induced ROS , probably through both of these activities . Sodium salicylate also has antiviral effects as the result of its potent antioxidant properties . Furthermore , by reducing ROS , aspirin and sodium salicylate inhibit CMV-induced NFkappaB activation , the ability of IE72 to transactivate its promoter , CMV IE gene expression after infection of SMCs , and CMV replication in SMCs . This is the first time aspirin has been shown to have antiviral effects . Thus , it is possible that aspirin has previously unrecognized therapeutic effects in various clinical situations , such as in viral infections ( when used as an antipyretic agent ) and in atherosclerosis ( when used as an antiplatelet agent ) . 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 . 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 . Effects of aspirin on atherosclerosis and the cyclooxygenase-2 expression in atherosclerotic rabbits . BACKGROUND : Atherosclerosis is a complex vascular inflammatory disease . DB00945 is a mainstay in the prevention of vascular complications of atherosclerosis . In this study , the effectiveness of aspirin in suppressing atherosclerosis and the inflammation process was evaluated in rabbits fed with a high fat diet . METHODS : Eighteen male New Zealand rabbits were randomly divided into 3 groups : control group , untreated cholesterol-fed group , aspirin treated cholesterol-fed group , which were fed for 12 weeks . After 12 weeks , the aorta was harvested for pathologic morphology observation . Immunohistochemical analysis of cyclooxygenase-2 ( P35354 ) , macrophage and vascular smooth muscle cell ( VSMC ) was performed . The statistical analysis was performed by the statistical program SPSS10.0 . RESULTS : The aorta plaque/intima size ( P/I ) by pathologic morphology observation was 0 % , ( 59.6 +/- 13.7 ) % and ( 36.3 +/- 16.5 ) % in the control , untreated cholesterol-fed group and aspirin treated group , respectively . The maximum plaque thickness , the degree of artery stenosis and the proportion of the intimal circumference occupied by atheroma of the 3 groups were significantly different from each other ( P < 0.01 ) . The expression of P35354 and macrophage in plaque of the aspirin treated group were decreased compared with that in untreated cholesterol-fed group . However , no difference was found in the expression of VSMC between the aspirin treated and the untreated cholesterol-fed group . CONCLUSION : The mechanism of atherosclerosis suppression by aspirin in cholesterol-fed rabbits is related to the inhibition of P35354 expression together with the reduced inflammation followed by , but not related to the hypolipidemic effects . 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 . DB00945 -responsive , migraine-like transient cerebral and ocular ischemic attacks and erythromelalgia in O60674 -positive essential thrombocythemia and polycythemia vera . Migraine-like cerebral transient ischemic attacks ( MIAs ) and ocular ischemic manifestations were the main presenting features in 10 O60674 (V617F)-positive patients studied , with essential thrombocythemia ( ET ) in 6 and polycythemia vera ( PV ) in 4 . Symptoms varied and included cerebral ischemic attacks , mental concentration disturbances followed by throbbing headaches , nausea , vomiting , syncope or even seizures . MIAs were frequently preceded or followed by ocular ischemic events of blurred vision , scotomas , transient flashing of the eyes , and sudden transient partial blindness preceded or followed erythromelalgia in the toes or fingers . The time lapse between the first symptoms of aspirin-responsive MIAs and the diagnosis of ET in 5 patients ranged from 4 to 12 years . At the time of erythromelalgia and MIAs , shortened platelet survival , an increase in the levels of the platelet activation markers β-thromboglobulin and platelet factor 4 and also in urinary thromboxane B2 were clearly indicative of the spontaneous in vivo platelet activation of constitutively O60674 (V617F)-activated thrombocythemic platelets . DB00945 relieves the peripheral , cerebral and ocular ischemic disturbances by irreversible inhibition of platelet cyclo-oxygenase ( P23219 ) activity and aggregation ex vivo . Vitamin K antagonist , dipyridamole , ticlopidine , sulfinpyrazone and sodium salicylate have no effect on platelet P23219 activity and are ineffective in the treatment of thrombocythemia-specific manifestations of erythromelalgia and atypical MIAs . If not treated with aspirin , ET and PV patients are at a high risk of major arterial thrombosis including stroke , myocardial infarction and digital gangrene . 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 . DB00435 mediates protective effect of endothelin receptor antagonism during myocardial ischemia and reperfusion . Endothelin ( ET ) receptor antagonism protects from ischemia-reperfusion injury . We hypothesized that the cardioprotective effect is related to nitric oxide ( NO ) bioavailability . Buffer-perfused rat and mouse hearts were subjected to ischemia and reperfusion . At the onset of ischemia , the rat hearts received vehicle , the dual endothelin type A/type B ( P25101 /ETB ) receptor antagonist DB00559 ( 10 microM ) , the NO synthase inhibitor NG-monomethyl-L-arginine ( L-NMMA ; 100 microM ) , the combination of DB00559 and L-NMMA or the combination of DB00559 , L-NMMA , and the NO substrate L-arginine ( 1 mM ) . Hearts from wild-type and endothelial NO synthase ( P29474 ) -deficient mice received either vehicle or DB00559 . Myocardial performance , endothelial function , NO outflow , and P29474 expression were monitored . DB00559 significantly improved myocardial function during reperfusion in rats and in wild-type mice , but not in P29474 -deficient mice . The functional protection afforded by DB00559 was inhibited by L-NMMA , whereas it was restored by L-arginine . Myocardial expression of P29474 ( immunoblotting ) increased significantly in DB00559 -treated rat hearts compared with vehicle hearts . Recovery of NO outflow during reperfusion was enhanced in the DB00559 -treated rat heart . The endothelium-dependent vasodilator adenosine diphosphate increased coronary flow by 18 +/- 9 % at the end of reperfusion in the DB00559 group , whereas it reduced coronary flow by 7 +/- 5 % in the vehicle group ( P < 0.001 ) . The response to the endothelium-independent dilator sodium nitroprusside was not different between the two groups . In conclusion , the dual P25101 /ETB receptor antagonist DB00559 preserved endothelial and cardiac contractile function during ischemia and reperfusion via a mechanism dependent on endothelial NO production . 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 . 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 . The role of cyclooxygenase in gastric mucosal protection . P23219 and P35354 are two cyclooxygenase enzymes responsible for prostanoid production . P35354 is expressed in inflammatory cells and fibroblasts of the gastric mucosa , and through the production of various growth factors including hepatocyte growth factor ( P14210 ) and vascular endothelial growth factor ( P15692 ) , plays a key role in the tissue repair process . DB00945 induces and acetylates P35354 to produce 15-(R)-epi-lipoxinA4 , an anti-inflammatory mediator thought to protect the gastric mucosa against aspirin-induced injury . Recently , three different PGE synthases have been identified , that convert P35354 metabolites into DB00917 . mPGE synthase ( mPGES ) -1 has been shown to be inducible , and to colocalize with P35354 in fibroblasts and macrophages infiltrating the gastric ulcer bed . Q15185 and Q9H7Z7 have been found expressed in normal gastric mucosa , with no change in expression levels seen in gastritis or gastric ulcer tissue . Finally , this review discusses the role of these enzymes in the pathophysiology of the gastric mucosa , as well as the biologcal significance of their inhibition .

MH_train_1144

MH_train_1144

MH_train_1144

interacts_with DB00243?

multiple_choice

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 . Presynaptic monoaminergic vesicles in Parkinson 's disease and normal aging . We present development and human application of a method for determining the regional cerebral density of the type 2 vesicular monoamine transporter ( Q05940 ) using positron emission tomography ( PET ) and [11C]dihydrotetrabenazine ( DTBZ ) . Previous animal studies indicate striatal Q05940 density is linearly related to the integrity of substantia nigra dopamine neurons and is not subject to drug- or lesion-compensatory regulation . In the present studies , kinetic compartmental modeling was employed to estimate blood-brain [11C]DTBZ transport ( P04264 ) and Q05940 binding site density ( tissue-to-plasma DTBZ distribution volume , DV ) from the cerebral and plasma DTBZ time courses after intravenous tracer injection . In controls , we found reductions of putamen DTBZ DVwith advancing age , corresponding to losses of 0.77 % per year in specific Q05940 binding . Parkinson 's disease ( PD ) patients had reduction in specific DTBZ DV in the putamen ( -61 % ) and in the caudate nucleus ( -43 % ) . There was no overlap of lowest specific putamen DTBZ DV between individual elderly controls and PD patients . The present results indicate the suitability of [11C]DTBZ PET for objective quantification of nigrostriatal integrity , including evaluation of PD progression and its possible therapeutic modification . 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 . DNA damage and S phase arrest induced by Ochratoxin A in human embryonic kidney cells ( P29320 293 ) . Ochratoxin A ( OTA ) is a ubiquitous mycotoxin with potential nephrotoxic , hepatotoxic and immunotoxic effects . The mechanisms underlying the nephrotoxicity of OTA remain obscure . To investigate DNA damage and the changes of the cell cycle distribution induced by OTA , human embryonic kidney cells ( P29320 293 cells ) were incubated with various concentrations of OTA for 24h in vitro . The results indicated that OTA treatment led to the production of reactive oxygen species ( ROS ) and to a decrease of the mitochondrial membrane potential ( ΔΨm ) . OTA-induced DNA damage in P29320 293 cells was evidenced by DNA comet tails formation and increased expression of γ- P16104 . In addition , OTA could induce cell cycle arrest at the S phase in P29320 293 cells . The expression of key cell cycle regulatory factors that were critical to the S phase , including cyclin A2 , cyclin E1 , and P24941 , were further detected . The expression of cyclin A2 , cyclin E1 , and P24941 were significantly decreased by OTA treatment at both the mRNA and protein levels . The apoptosis of P29320 293 cells after OTA treatment was observed using Hoechst 33342 staining . The results confirmed that OTA did induce apoptosis in P29320 293 cells . In conclusion , our results provided new insights into the molecular mechanisms by which OTA might promote nephrotoxicity . Pituitary-specific and hormonally regulated gene expression directed by the rat proopiomelanocortin promoter in transgenic mice . All aspects of P01189 biosynthesis exhibit tissue-specific regulation . The single copy gene is highly expressed in anterior lobe ( AL ) corticotrophs and intermediate lobe ( IL ) melanotrophs of the pituitary gland and in the arcuate nucleus of the hypothalamus . P01189 gene transcription in corticotrophs is induced by hypothalamic P06850 and vasopressin and inhibited by adrenal glucocorticoids , while in melanotrophs it is predominantly regulated by beta-adrenergic neural input and dopamine . To identify the rat P01189 ( rPOMC ) gene sequences necessary and sufficient to target expression and hormonal regulation in corticotrophs and melanotrophs , we generated 13 transgenic mice carrying rPOMC fusion genes . The genes consisted of 706 or 480 basepairs of rPOMC 5' flanking sequences ligated to either the E. coli LacZ gene encoding beta-galactosidase or the P04264 mutant of the SV40 large T-antigen gene . Overall , half of the transgenic lines had reporter gene expression in their AL and IL in a pattern indistinguishable from DB01285 immunohistochemistry . In three of these lines , beta-galactosidase or P04264 T-antigen was localized by double immunofluorescence exclusively to DB01285 -positive corticotrophs and melanotrophs . Transcriptional regulation of the rPOMC-LacZ fusion gene in response to hormonal manipulation was quantified by a fluorescence assay for beta-galactosidase enzyme activity in pituitary extracts . There was a 15-fold increase in AL enzyme activity after adrenalectomy and a 3-fold increase in IL activity after haloperidol treatment . X-gal histochemistry of pituitaries from hormonally treated mice confirmed the cellular specificity of these effects. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . 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 . 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 . Activation of O14788 -induced osteoclasts and memory T lymphocytes by Porphyromonas gingivalis is serotype dependant . AIM : Destructive periodontitis is associated with a Th1-Th17 immune response and activation of O14788 -induced osteoclasts . In addition , Porphyromonas gingivalis P04264 and K2 serotypes induce a strong Th1-Th17 response . This study aimed to investigate whether these P. gingivalis serotypes induce higher osteoclasts activation , by increased Th17-associated O14788 production , and an antigen-specific memory T-lymphocyte response . MATERIAL AND METHODS : The O14788 production and TRAP(+) osteoclast induction were quantified on naïve T lymphocytes stimulated with dendritic cells primed with the P. gingivalis serotypes . The T-bet , GATA-3 , RORC2 and Foxp3 expression was correlated with O14788 production . The frequency of proliferating memory T lymphocytes in response to P. gingivalis serotypes was determined in both periodontitis and healthy subjects . RESULTS : T lymphocytes stimulated by P04264 or K2-primed dendritic cells elicited higher levels of O14788 and TRAP(+) osteoclasts than cells stimulated with the other serotypes . O14788 positively correlated with RORC2 . Whereas periodontitis patients had a higher frequency of memory T lymphocytes responding to P04264 or K2 , healthy subjects had a higher frequency of memory T lymphocytes responding to P19013 or K(-) . CONCLUSIONS : P. gingivalis serotypes P04264 and K2 , but not others , are associated with an increased production of the osteoclastogenesis-related factor O14788 . This important information suggests that these serotypes could elicit a greater bone resorption in vivo and have a role in the periodontitis pathogenesis . 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 . In vitro evaluation of Cordyceps militaris as a potential radioprotective agent . Radiation is an important component of therapy for a wide range of malignant conditions . However , it triggers DNA damage and cell death in normal cells and results in adverse side-effects . Cordyceps militaris ( C. militaris ) , a traditional medicinal mushroom , produces the bioactive compound , cordycepin ( 3'-deoxyadenosine ) and has multiple pharmacological activities , such as antitumor , antimetastatic , antioxidant and immunomodulatory effects . The present study was undertaken to investigate whether CM-AE , an extract obtained from C. militaris exerts protective effects against radiation-induced DNA damage . The protective effects of CM-AE were compared with those of cordycepin . CM-AE effectively increased free radical scavenging activity and decreased radiation-induced plasmid DNA strand breaks in in vitro assays . CM-AE significantly inhibited the generation of reactive oxygen species ( ROS ) and cellular DNA damage in 2 Gy irradiated Chinese hamster ovary ( CHO ) - P04264 cells . Moreover , treatment with CM-AE induced similar levels of phosphorylated P16104 in the cells , which reflects the initial DNA double-strand breaks in the irradiated cells compared with the non-irradiated CHO- P04264 cells . However , cordycepin did not show free radical scavenging activity and did not protect against radiation-induced plasmid DNA or cellular DNA damage . These results suggest that the free radical scavenging activity of CM-AE contributes towards its DNA radioprotective effects and that the protective effects of CM-AE are much more potent to those of cordycepin . The data presented in this study may provide useful information for the screening of potent radioprotective materials . β-Arrestin-1 mediates thyrotropin-enhanced osteoblast differentiation . DB00024 ( DB00024 ) activation of the DB00024 receptor ( P16473 ) , a 7-transmembrane-spanning receptor ( 7TMR ) , may have osteoprotective properties by direct effects on bone . P16473 activation by DB00024 phosphorylates protein kinases P31749 , p38α , and P27361 /2 in some cells . We found DB00024 -induced phosphorylation of these kinases in 2 cell lines engineered to express TSHRs , human embryonic kidney P29320 - P16473 cells and human osteoblastic U2OS- P16473 cells . In U2OS- P16473 cells , DB00024 up-regulated pAKT1 ( 7.1±0.5-fold ) , p38α ( 2.9±0.4-fold ) , and pERK1/2 ( 3.1±0.2-fold ) , whereas small molecule P16473 agonist P06681 had no or little effect on pAKT1 ( 1.8±0.08-fold ) , p38α ( 1.2±0.09-fold ) , and pERK1/2 ( 1.6±0.19-fold ) . Furthermore , DB00024 increased expression of osteoblast marker genes P05186 ( 8.2±4.6-fold ) , O14788 ( 21±5.9-fold ) , and osteopontin ( P10451 ; 17±5.3-fold ) , whereas P06681 had little effect ( P05186 , 1.7±0.5-fold ; O14788 , 1.3±0.6-fold ; and P10451 , 2.2±0.7-fold ) . β-Arrestin-1 and -2 can mediate activatory signals by 7TMRs . DB00024 stimulated translocation of β-arrestin-1 and -2 to P16473 , whereas P06681 failed to translocate either β-arrestin . Down-regulation of β-arrestin-1 by siRNA inhibited DB00024 -stimulated phosphorylation of P27361 /2 , p38α , and P31749 , whereas down-regulation of β-arrestin-2 increased phosphorylation of P31749 in both cell types and of P27361 /2 in P29320 - P16473 cells . Knockdown of β-arrestin-1 inhibited DB00024 -stimulated up-regulation of mRNAs for P10451 by 87 ± 1.7 % and O14788 by 73 ± 2.4 % , and P10451 secretion by 74 ± 10 % . We conclude that DB00024 enhances osteoblast differentiation in U2OS cells that is , in part , caused by activatory signals mediated by β-arrestin-1 . Human and mouse trace amine-associated receptor 1 have distinct pharmacology towards endogenous monoamines and imidazoline receptor ligands . TAARs ( trace amine-associated receptors ) are G-protein-coupled receptors that respond to low abundance , endogenous amines such as tyramine and tryptamine , and represent potential targets for neuropsychiatric diseases . However , some members of this receptor subfamily either have no ligand identified or remain difficult to express and characterize using recombinant systems . In the present paper we report the successful expression of human and mouse Q96RJ0 , and the characterization of their responses to various natural and synthetic agonists . In P29320 ( human embryonic kidney ) -293/CRE-bla cells , mouse Q96RJ0 showed a robust response to trace amines as measured using either a DB02527 assay or a beta-lactamase reporter assay , whereas human Q96RJ0 showed a weaker , but still measurable , response . When certain fragments of human Q96RJ0 were replaced with the corresponding regions of mouse Q96RJ0 , the chimaeric receptor showed a much stronger response in DB02527 production . Examination of a series of agonists on these receptors revealed that the human and the chimaeric receptor are almost identical in pharmacology , but distinct from the mouse receptor . We also screened small libraries of pharmacologically active agents on Q96RJ0 and identified a series of synthetic agonists , some of which are also ligands of the enigmatic imidazoline receptor . The findings of the present study not only shed light on the pharmacological species difference of Q96RJ0 , but also raise new possibilities about the mechanism of some of the imidazoline-related agents . A replication-competent adenovirus assay for E1-deleted Ad35 vectors produced in O15534 . P13671 cells . The presence of replication-competent adenovirus ( RCA ) is a safety concern for biologics based on recombinant adenoviruses and RCA testing is therefore mandatory for release of clinical material . RCA , which arises from homologous recombination between Ad5 vectors and P29320 -293 cells , can be eliminated by the use of O15534 . P13671 cells in combination with a matched vector . However , little is known on RCA formation with vectors based on adenovirus serotypes other than Ad5 and reliable RCA assays to test them are generally lacking . Here we report on the development and qualification of a sensitive RCA assay for Ad35 , a promising alternative to Ad5 vectors . The assay is able to detect 1 RCA in 3x10(10) vector particles with 95 % confidence , thus meeting current FDA requirements , and can discriminate between RCA and other rare P16870 -causing entities , including helper dependent E1 positive particles ( HDEP ) . Using this assay , the first batches of Ad35 vectors produced in O15534 . P13671 cells were analysed and found to be free of RCA and HDEP . Based on the statistical model used , we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors . The P55008 -phase growth-arresting action of interleukin-1 is independent of p53 and P38936 / P38936 function . Interleukin-1 ( IL-1 ) causes P55008 -phase growth arrest of A375- P13671 human melanoma cells by hypophosphorylation of the retinoblastoma susceptibility gene product Rb . Because p53 and P38936 / P38936 proteins are key components of growth arrest pathways involving Rb hypophosphorylation , we tested the functional role of these two proteins in IL-1 action . Exposure to IL-1 caused induction of both p53 and P38936 / P38936 proteins . However , inhibition of p53 function by the P04264 mutant of SV40-T antigen or by m175 ( DB00125 to DB00117 ) dominant-negative mutant of p53 did not result in abrogation of IL-1 action , suggesting that p53 function is not required for growth arrest by IL-1 . Studies aimed at testing the role of P38936 / P38936 in IL-1 action indicated that IL-1 induced P38936 / P38936 expression independently of the p53 status of the cells . However , inhibition of P38936 / P38936 expression resulted in only a marginal rescue from the growth-arresting action of IL-1 . These findings imply that despite their induction , neither wild-type p53 nor P38936 can fully account for the growth arrest by IL-1 . Thus , a p53- and P38936 -independent pathway(s) mediates IL-1 action . 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 . Functional characterization of a novel serotonin receptor ( 5-HTap2 ) expressed in the CNS of Aplysia californica . Serotonin has been shown to be a neuromodulator in the Aplysia californica CNS . The diversity of serotonin actions is due to the existence of several different receptor subtypes . In this study we report the cloning of a full-length cDNA , coding for a novel serotonin receptor ( 5-HTap2 ) . The receptor protein bears the characteristics of G protein-coupled receptors . It shares 68 % and 34 % of its amino acid sequence identity with the 5-HTlym receptor from Lymnaea stagnalis and the mammalian P08908 receptor , respectively . When transfected in P29320 293 cells , 5-HTap2 was negatively coupled to adenylate cyclase . Ligand binding analysis indicated that the order of potencies of various drugs for the inhibition of [3H]LSD binding was : methiothepin > metergoline > 5-CT > PAPP > 5-HT > ketanserin > NAN-190 > 8-OH-DPAT > clozapine . RT-PCR amplification of RNA isolated from different tissues indicated that this receptor is expressed in the CNS and in bag cells . The expression of 5-HTap2 restricted to the CNS suggests an important role for this receptor in the modulation of neuronal functions in Aplysia . Moreover , the high expression of 5-HTap2 in the bag cells , associated with its pharmacological profile , suggests that this receptor may be implicated in modulating the afterdischarge during the egg-laying behavior . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . 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 . 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 . 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 . 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 . 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 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 . 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 . 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 .

MH_train_1145

MH_train_1145

MH_train_1145

interacts_with DB08895?

multiple_choice

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 . 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 . [ Genetic polymorphisms and cancer risk ] . While hereditary disease genes have a high lifelong cumulative incidence rate ( penetrance ) , the penetrance for polymorphism genotypes is not high . Polymorphisms relating to cancer incidence are classified into 1. carcinogen metabolizing enzymes ( CYPs , GSTs , P15559 , etc. ) , 2 . DNA repair enzymes ( O15527 , P18887 , P18074 , etc. ) , 3 . DNA synthesis and methylation ( P42898 , MS , etc. ) , 4. cytokines and inflammation-related enzymes ( IL-1B , P01375 -A , P05164 , etc. ) , and 5. sex hormone metabolizing enzymes and the receptors ( P11511 , P31213 , ER , etc. ) . Since genotypes can not be manipulated , they are not the factors subject to prevention . However , the finding that the strength of association between lifestyle and disease occurrence is influenced by genotypes ( gene-environment interaction ) , opens the door to genotype applications for disease prevention practice . 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 . Repurposing of metformin and aspirin by targeting AMPK- P42345 and inflammation for pancreatic cancer prevention and treatment . Pancreatic cancer , as the fourth leading cause of cancer-related deaths , carries a poor prognosis with a median survival of 6 months and a dismal 5-year survival rate of 3 % to 5 % . These statistics highlight an urgent need for novel chemopreventive and therapeutic strategies for this malignancy . Metformin and aspirin have been explored as two emerging cancer chemoprevention agents for different types of cancers , including pancreatic cancer . Here , we review the effects of both metformin and aspirin on pancreatic tumorigenesis and their potential actions in pancreatic cancer . Special attention is paid to their effects on the important signaling pathways of pancreatic cancer development as well as possible mechanisms for synergy between these two agents . For metformin , the most important mechanism may involve the inhibition of P42345 signaling via AMP-activated protein kinase ( AMPK ) -dependent and -independent pathways . For aspirin , the major mechanism is the anti-inflammatory action through the inhibition of P23219 / P35354 and modulation of the NFκB or P40763 pathway . In addition , aspirin may activate AMPK , and both agents may affect Notch , Wnt/β-catenin , and other signaling pathways . The combination of metformin and aspirin will provide additive and possibly synergistic effects for the prevention and treatment of pancreatic cancer . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Genetic profiling of myeloproliferative disorders by single-nucleotide polymorphism oligonucleotide microarray . OBJECTIVE : Myeloproliferative disorders ( P53602 ) are clonal hematopoietic diseases that include polycythemia vera ( PV ) , essential thrombocytosis ( ET ) , and primary myelofibrosis ( PMF ) . Mutations in O60674 are present in many P53602 patients . Additional genomic abnormalities are not fully examined in P53602 . MATERIALS AND METHODS : We used single-nucleotide polymorphism DNA microarray ( SNP-chip ) to analyze 43 patients with P53602 ( 10 PV , 17 ET , and 16 PMF ) for genomic aberrations . RESULTS : Genomic abnormalities were rare in ET . The region containing either RB ( 13q14 ) or P21359 ( 17q11 ) was deleted in 4 of the 16 PMF , especially PMF with no O60674 mutations . All five cases of PV having homozygous JAK2V617F had loss of heterozygosity with normal copy number [ uniparental disomy ] involving the gene . A subpopulation with 9p uniparental disomy was detected in 11 P53602 ( 3 PV , 1 ET , 7 PMF ) . Uniparental disomy at 1p was found in one PV and three PMF . A novel mutation of P40238 ( Y591D ) , which was involved in this uniparental disomy , was found in 1 PV with O60674 mutation . The other three cases of PMF with 1p uniparental disomy had point mutations of the P40238 gene , either a novel mutation ( S204F ) or the previously described W515L . CONCLUSION : Genomic abnormalities , including 9p uniparental disomy/ O60674 point mutations , 1p uniparental disomy/ P40238 point mutations , deletions of P06400 and P21359 are common alterations in P53602 , especially in PMF . 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 . 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 . Q8WXI7 induced rapid G2/M transition via interactions with O60674 for increased proliferation and anti-apoptosis in breast cancer cells . Q8WXI7 / Q8WXI7 is a tumor marker currently used in clinics for the follow-up of patients with ovarian cancer . However , Q8WXI7 expression is not entirely restricted to ovarian malignancies and has been reported in other cancers including breast cancer . Although it is well established as a biomarker , function of Q8WXI7 in cancer remains to be elucidated . In the present study , we investigated the role of Q8WXI7 in breast cancer and its underlying mechanisms . Interestingly , our results showed that Q8WXI7 is overexpressed in breast cancer tissues whereas not expressed in non-neoplastic ducts . Further , stable knockdown of Q8WXI7 in breast cancer cells ( MDA MB 231 and HBL100 ) resulted in significant decrease in the rate of cell growth , tumorigenicity and increased apoptosis . In search of a mechanism for breast cancer cell proliferation we found that Q8WXI7 interacts with the ezrin/radixin/moesin domain-containing protein of Janus kinase ( O60674 ) as demonstrated by the reciprocal immunoprecipitation method . These interactions mediate phosphorylation of P40763 ( Tyr705 ) , which might be a potential mechanism for Q8WXI7 -induced proliferation of breast cancer cells by a subsequent co-transactivation of transcription factor c-Jun . Furthermore , silencing of Q8WXI7 induced G2/M arrest in breast cancer cells through downregulation of P12004 B1 and decreased phosphorylation of O14965 . This in turn led to enhanced apoptosis in the Q8WXI7 -knockdown breast cancer cells through P01375 -related apoptosis-inducing ligand ( P50591 ) -mediated extrinsic apoptotic pathway with the help of c-Jun N-terminal kinase signaling . Collectively , our results suggest that Q8WXI7 has a dual role in breast cancer cell proliferation by interacting with O60674 and by inhibiting the apoptotic process through downregulation of P50591 . 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 . 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 . 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 . 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 . 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 . 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 . Basal cell carcinoma and variants in genes coding for immune response , DNA repair , folate and iron metabolism . Basal cell carcinoma ( BCC ) is one of the most common neoplasms in the world and its incidence has been increasing worldwide in recent years . BCCs are caused by an interplay between genetic and environment factors . We conducted a case-control association study in BCC patients and controls from Sweden and Finland . Fifteen single nucleotide polymorphisms ( SNPs ) , P05231 -174G/C , -634G/C , and -597G/A ; P22301 -1082G/A and -592C/A ; IL-1beta-511C/T ; NBS1 exon 5 Glu185Gln ; Q01831 exon 15 Lys939Gln ; P18074 exon 23 Lys751Gln ; P18887 exon 10 Arg399Gln ; O43542 exon 7 Thr241Met ; cyclin D1 exon 4 G870A ; P42898 exon 4 Ala222Val and exon 7 Glu429Ala ; Q30201 exon 4 C282Y were performed by Pyrosequencing and RFLP techniques . Most of the genotype distributions were in accordance with the Hardy-Weinberg equilibrium ( HWE ) , except for P22301 -1082G/A , where cases with BCC showed a significant deviation from HWE ( P = 0.04 ) . Linkage disequilibrium was observed between the -174 and -597 alleles in the P05231 gene in the present populations . No difference between BCC and controls appeared in any of the SNPs analyzed . Only the combined distributions of TT/AA genotypes in P42898 exon 4 ( C/T ) and exon 7 ( A/C ) showed slight increase in BCC compared to controls ( P < 0.07 , OR : 1.94 ; 95 % CI : 0.96-3.89 ) . 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 . JAK- P35610 and JAK-PI3K-mTORC1 pathways regulate telomerase transcriptionally and posttranslationally in ATL cells . Adult T-cell leukemia ( ATL ) is a heterogeneous tumor that is resistant to chemotherapy . Telomerase activity plays a critical role in tumorigenesis and is associated with the prognosis of ATL patients . Interleukin ( IL ) -2 commonly promotes tumor growth in chronic ATL cells . The signaling pathways involved in P60568 -regulated telomerase activation were studied in ATL cells derived from chronic ATL patients . P60568 challenge enhanced tyrosine phosphorylation of Janus-activated kinase (JAK)1-3 and P42229 , and induced P23458 and O60674 to associate with P42229 in P60568 -dependent ATL cells . Chromatin immunoprecipitation assays revealed that P42229 directly bound to the human telomerase reverse transcriptase ( hTERT ) promoter . P42229 short interfering RNA inhibited hTERT transcription in P60568 -stimulated ATL cells . Inhibitors of PI3K , HSP90 , and P42345 reduced P60568 -induced hTERT mRNA , protein expression , and telomerase activity . AKT , HSP90 , P42345 , S6 kinase , and hTERT immunoprecipitate from P60568 -stimulated cells contained telomerase activity , suggesting that hTERT directly interacts with , and is regulated by , these proteins . Binding of the p85 regulatory subunit of PI3K to O60674 was enhanced in an P60568 -dependent manner , indicating that O60674 propagates activation signals from the P60568 receptor and links hTERT activation to both the P42229 and PI3K pathways . Finally , P60568 -induced activation of telomerase and P42229 was observed in primary leukemic cells . These results indicate that P60568 stimulation induces hTERT activation through the JAK/ P35610 pathway and the JAK/PI3K/AKT/HSP90/mTORC1 pathway in P60568 -responsive ATL cells . These signaling proteins represent novel and promising molecular therapeutic targets for P60568 -dependent ATL . 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 . 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 . 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 . 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 . Human monocytes are severely impaired in base and DNA double-strand break repair that renders them vulnerable to oxidative stress . Monocytes are key players in the immune system . Crossing the blood barrier , they infiltrate tissues and differentiate into ( i ) macrophages that fight off pathogens and ( ii ) dendritic cells ( DCs ) that activate the immune response . A hallmark of monocyte/macrophage activation is the generation of reactive oxygen species ( ROS ) as a defense against invading microorganisms . How monocytes , macrophages , and DCs in particular respond to ROS is largely unknown . Here we studied the sensitivity of primary human monocytes isolated from peripheral blood and compared them with macrophages and DCs derived from them by cytokine maturation following DNA damage induced by ROS . We show that monocytes are hypersensitive to ROS , undergoing excessive apoptosis . These cells exhibited a high yield of ROS-induced DNA single- and double-strand breaks and activation of the ATR-Chk1- Q13315 -Chk2-p53 pathway that led to Fas and caspase-8 , -3 , and -7 activation , whereas macrophages and DCs derived from them were protected . Monocytes are also hypersensitive to ionizing radiation and oxidized low-density lipoprotein . The remarkable sensitivity of monocytes to oxidative stress is caused by a lack of expression of the DNA repair proteins P18887 , ligase IIIα , poly(ADP-ribose) polymerase-1 , and catalytic subunit of DNA-dependent protein kinase ( DNA-PK(cs) ) , causing a severe DNA repair defect that impacts base excision repair and double-strand break repair by nonhomologous end-joining . During maturation of monocytes into macrophages and DCs triggered by the cytokines GM- P04141 and P05112 , these proteins become up-regulated , making macrophages and DCs repair-competent and ROS-resistant . We propose that impaired DNA repair in monocytes plays a role in the regulation of the monocyte/macrophage/DC system following ROS exposure . Mechanisms of Nattokinase in protection of cerebral ischemia . In vivo , the level of cyclic DB00640 Monophosphate ( DB02527 ) and the pathway of the Janus Kinase1/Signal Transducers and Activators of Transcription1 ( P23458 / P42224 ) were studied . In vitro , the Ca(2+) mobilization in human platelet stimulated by thrombin was observed . In addition , vasomotion of vascular smooth muscle was measured by adding DB00761 or norepinephrine(NE) under the Ca(2+) contained bath solutions . The effect induced by NE in the presence of N-nitro-L-arginine methyl ester ( L-NAME ) or indometacin ( Indo ) was also detected . At last , the levels of tissue plasminogen activator ( t-PA ) and P00747 activator inhibitor-1 ( P05121 ) in cultured supernatans in Human umbilical vein endothelial cells ( Huvecs ) were measured by means of ELISA kit . Results showed that Nattokinase ( NK ) significantly increased the DB02527 level , activated the signal passage of P23458 / P42224 in injured part and inhibited remarkably the rise of platelet intracellular Ca(2+) ( [Ca(2+)]i ) in human platelet . Furthermore , NK relaxed rat thoracic aortic artery in the dose-dependent manner and in the endothelium dependent manner and its effect could be attenuated by L-NAME . Also , the secretion of t-PA and P05121 were reduced stimulated by Adr on Huvecs . These data indicated that the neuroprotective effect of NK was associated with its antiplatelet activity by elevating DB02527 level and attenuating the calcium release from calcium stores ; with its anti-apoptotic effect through the activation of P23458 / P42224 pathway ; with its relaxing vascular smooth muscle by promoting synthesis and release of NO , reducing ROC calcium ion influx and with its protection on endothelial cells through increasing fibrinolytic activity and facilitating spontaneous thrombolysis . 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 . Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats . BACKGROUND : Sepsis and endotoxemia are associated with concurrent activation of inflammation and the hemostatic mechanism , which both contribute to organ dysfunction and death . Electrical vagus nerve stimulation ( VNS ) has been found to inhibit tumor necrosis factor ( P01375 ) -alpha release during endotoxemia in rodents . OBJECTIVE : To determine the effect of VNS on activation of coagulation and fibrinolysis . METHODS : Rats received a sublethal i.v. dose of lipopolysaccharide ( LPS ) after electrical VNS or sham stimulation . Activation of coagulation and fibrinolysis , as well as cytokine release , was measured before LPS injection and 2 , 4 and 6 h thereafter . RESULTS : LPS induced activation of the coagulation system ( increases in the plasma concentrations of thrombin-antithrombin complexes and D-dimer , and a decrease in antithrombin ) and biphasic changes in the fibrinolytic system [ early rises of plasminogen activator activity and tissue-type plasminogen activator , followed by a delayed increase in plasminogen activator inhibitor type 1 ( P05121 ) ] . VNS strongly inhibited all LPS-induced procoagulant responses and more modestly attenuated the fibrinolytic response . In addition , VNS attenuated the LPS-induced increases in plasma and splenic concentrations of the proinflammatory cytokines P01375 and interleukin-6 ( P05231 ) , while not influencing the release of the anti-inflammatory cytokine P22301 . CONCLUSION : These data illustrate a thus far unrecognized effect of VNS and suggest that the cholinergic anti-inflammatory pathway not only impacts on inflammation but also on the coagulant-anticoagulant balance . 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 . 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 . P49841 inhibitor suppresses Porphyromonas gingivalis lipopolysaccharide-induced P25942 expression by inhibiting nuclear factor-kappa B activation in mouse osteoblasts . Bone-forming osteoblasts have been recently reported capable of expressing the critical co-stimulatory molecule P25942 upon exposure to bacterial infection , which supports the unappreciated role of osteoblasts in modulating bone inflammation . Recent studies highlight the anti-inflammatory potential of glycogen synthase kinase-3β ( GSK-3β ) inhibitors ; however , their effect on osteoblasts remains largely unclear . In the present study , we showed that treatment with SB216763 , a highly specific GSK-3β inhibitor , resulted in a dose-dependent decrease in the mRNA and protein expression of P25942 , as well as production of pro-inflammatory cytokines P05231 , P01375 -α and IL-1β , in the Porphyromonas gingivalis-lipopolysaccharide ( LPS ) -stimulated murine osteoblastic-like MC3T3-E1 cells . Furthermore , inhibition of GSK-3β remarkably represses the LPS-induced activation of the nuclear factor kappa B ( NF-κB ) signaling pathway by suppressing IκBα phosphorylation , NF-κBp65 nuclear translocation , and NF-κBp65 DNA binding activity . Closer investigation by immunoprecipitation assay revealed that β-catenin can physically interact with NF-κBp65 . The negative regulation effect of GSK-3β inhibitor on P25942 expression is mediated through β-catenin , for siRNA of β-catenin attenuated the GSK-3β inhibitor-induced repression of NF-κB activation and , consequently , the expression of P25942 and production of pro-inflammatory cytokines in LPS-stimulated MC3T3-E1 cells . Thus our results elucidate the molecular mechanisms whereby GSK-3β inhibitor prevents the LPS-induced P25942 expression on osteoblasts and provide supportive evidence of the potential role of GSK-3β inhibitors in suppressing the immune function of osteoblasts in inflammatory bone diseases . Activation of the JAK- P35610 signaling pathway in the rat basilar artery after subarachnoid hemorrhage . The Janus kinase-signal transducer and activator of transcription ( JAK- P35610 ) is one of the most important signaling pathways transducing signals from the cell surface in response to cytokines . Subarachnoid hemorrhage ( Q53FZ2 ) produces cytokines in the P04141 . We investigated whether this signaling pathway is activated in the rat basilar artery after Q53FZ2 by cytokines . In a rat single-hemorrhage model of Q53FZ2 , basilar arteries and P04141 were obtained until 7 days after Q53FZ2 . The concentration of interleukin-6 ( P05231 ) in P04141 was measured by ELISA . Western blot analysis with P23458 , phosphospecific- P23458 , P40763 , phosphospecific P40763 at Tyr705 and Ser727 , cyclooxygenase-2 ( P35354 ) , and actin antibodies was performed in basilar artery . The expressions of P40763 , phosphospecific P40763 at Tyr705 and Ser727 , and P35354 in basilar artery were examined by immunohistochemical studies . The concentration of P05231 immediately increased after Q53FZ2 and Western blot analysis revealed that P23458 was phosphorylated within 2 h , accompanied by phosphorylation of P40763 at Tyr705 , extending to Ser727 at days 1-2 . Immunohistochemistry revealed phosphorylation of P40763 to occur in endothelial and smooth muscle cells of the basilar artery . In addition , intracisternal injection of P05231 by itself significantly increased phosphorylation of P40763 at Tyr705 and Ser727 . Expression of P35354 was also upregulated in endothelial cells of the basilar artery . These results indicate that Q53FZ2 produces the proinflammatory cytokine P05231 in the P04141 , which activates the JAK- P35610 signaling pathway in the basilar artery and induces transcription of immediate early genes . 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 . 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 . The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . 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 . Tumor Necrosis Factor alpha ( TNFalpha ) regulates P25942 expression through Q8N9N5 phosphorylation . P25942 plays an important role in mediating inflammatory response and is mainly induced by JAK/ P35610 phosphorylation cascade . TNFalpha is the key cytokine that activates P25942 during inflammation and tumorigenesis . We have earlier shown that Q8N9N5 can repress the transcription of P12004 D1 promoter by forming a Q13547 dependent repressor complex . In this study , we show that Q8N9N5 regulates the transcription of NF-kappaB target gene P25942 . Q8N9N5 recruits Q13547 and forms a repressor complex on P25942 promoter and keeps its basal transcription in check . Further , we show that TNFalpha stimulation induces Q8N9N5 phosphorylation at DB00133 -347 and promotes its cytoplasmic translocation , thus releasing its negative effect . Concomitantly , TNFalpha induced phosphorylation of P42224 at DB00135 -701 by P23458 facilitates its nuclear translocation and activation of P25942 through p300 recruitment and core Histone-3 acetylation . Thus , TNFalpha mediated regulation of P25942 expression occurs by dual phosphorylation of Q8N9N5 and P42224 . 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 . Exposure to low doses of formaldehyde during pregnancy suppresses the development of allergic lung inflammation in offspring . DB03843 ( FA ) is an environmental and occupational pollutant , and its toxic effects on the immune system have been shown . Nevertheless , no data are available regarding the programming mechanisms after FA exposure and its repercussions for the immune systems of offspring . In this study , our objective was to investigate the effects of low-dose exposure of FA on pregnant rats and its repercussion for the development of allergic lung inflammation in offspring . Pregnant Wistar rats were assigned in 3 groups : P ( rats exposed to FA ( 0.75 ppm , 1 h/day , 5 days/week , for 21 days ) ) , C ( rats exposed to vehicle of FA ( distillated water ) ) and B ( rats non-manipulated ) . After 30 days of age , the offspring was sensitised with ovalbumin ( OVA ) -alum and challenged with aerosolized OVA ( 1 % , 15 min , 3 days ) . After 24 h the OVA challenge the parameters were evaluated . Our data showed that low-dose exposure to FA during pregnancy induced low birth weight and suppressed the development of allergic lung inflammation and tracheal hyperresponsiveness in offspring by mechanisms mediated by reduced anaphylactic antibodies synthesis , P05231 and P01375 secretion . Elevated levels of P22301 were found . Any systemic alteration was detected in the exposed pregnant rats , although oxidative stress in the uterine environment was evident at the moment of the delivery based on elevated P23219 expression and reduced P29474 and SOD-2 in the uterus . Therefore , we show the putative programming mechanisms induced by FA on the immune system for the first time and the mechanisms involved may be related to oxidative stress in the foetal microenvironment . 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 . 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 ) 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 . 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 . 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 . 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 . 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 . 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 . 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 . P05231 and soluble P05231 receptor stimulate the production of MMPs and their inhibitors via JAK- P35610 and P29323 -MAPK signalling in human chondrocytes . Elevated concentrations of P05231 ( interleukin-6 ) and sIL-6r ( soluble P05231 receptor ) in the synovial fluid and serum of patients with arthritis have been implicated in joint cartilage destruction . This study examined the effects of P05231 and sIL-6r on the expression of MMPs ( matrix metalloproteinases ) , TIMPs ( tissue inhibitor of metalloproteinases ) , the plasminogen activation system including tPA ( tissue-type PA ) , uPA ( urokinase-type PA ) and P05121 ( PA inhibitor type 1 ) using chondrocytes derived from normal human femur cartilage . The cells were cultured with or without 50 ng/ml P05231 and/or 30 ng/ml sIL-6r in the presence or absence of the P52333 ( P52333 ) inhibitor WHI-P131 or the MEK [ MAPK ( mitogen-activated protein kinase ) / P29323 ( extracellular signal protein kinase ) kinase ] inhibitor PD98059 for up to 28 days . The expression of MMPs , TIMPs , uPA , tPA and P05121 was investigated at the mRNA and protein levels . MMP protein expression and pSTAT3 ( phosphorylation of signal transducer and activator of transcription 3 ) and pERK ( phosphorylation of P29323 ) were also measured . Treatment with both P05231 and sIL-6r markedly increased the expression of P03956 , P45452 , P01033 and P05121 , while significantly decreasing the expression of tPA and uPA and stimulating pSTAT3 and pERK . Adding WHI-P131 or PD98059 decreased P05231 and sIL-6r enhancement of P03956 , -3 and -13 . The results suggest that P05231 and sIL-6r stimulate the production of MMPs and their inhibitor via JAK- P35610 and P29323 -MAPK signalling in chondrocytes . A fusion cytokine coupling P04141 to P15248 induces heterologous receptor clustering and P42224 hyperactivation through O60674 promiscuity . Cytokine receptors are randomly distributed on the cell surface membrane and are activated upon binding of their extracellular ligands to mediate downstream cellular activities . We hypothesized that pharmaceutical clustering of ligand-bound , activated receptors may lead to heretofore unrealized gain-of-function with therapeutically desirable properties . We here describe an engineered bifunctional cytokine borne of the fusion of Granulocyte Macrophage Colony Stimulating Factor ( P04141 ) and P15248 ( P15248 ) ( hereafter GIFT9 fusokine ) and demonstrate that it chaperones co-clustering of the functionally unrelated P04141 receptor ( GMCSFR ) and P15248 receptor ( Q01113 ) on cell surface of target cells . We demonstrate that GIFT9 treatment of MC/9 cells leads to transhyperphosphorylation of Q01113 -associated P42224 by GMCSFR-associated O60674 . We also show that Q01113 -associated P23458 and P52333 augment phosphorylation of GMCSFR-linked P42229 . The functional relevance of these synergistic JAK/ P35610 transphosphorylation events translates to an increased mitogenic response by GMCSFR/ Q01113 -expressing primary marrow mast cells . The notion of inducing heterologous receptor clustering by engineered fusokines such as GIFT9 opens the door to a novel type of biopharmaceutical platform where designer fusokines modulate cell physiology through clustering of targeted receptor complexes . 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 . 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 . 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 . Enhancement of placenta growth factor expression by oncostatin M in human rheumatoid arthritis synovial fibroblasts . Oncostatin M ( P13725 ) belongs to P05231 subfamily and is mostly produced by T lymphocytes . High levels of P13725 are detected in the pannus of rheumatoid arthritis ( RA ) patients and it may arouse the inflammation responses in joints and eventually leads to bone erosion . P49763 ( P49763 ) is an angiogenic factor and highly homologous with vascular endothelial growth factor ( P15692 ) . It has been recently reported that P49763 is highly expressed in synovial tissue and enhances the production of proinflammatory cytokines including P01375 -α and P05231 . Here , we demonstrated that P13725 increased mRNA and protein levels of P49763 in a time- and concentration-dependent manner in RA synovial fibroblasts . Inhibitors of P52333 and PI3K antagonized P13725 -induced production of P49763 . P13725 enhanced the phosphorylation of Tyr705- P40763 , Ser727- P40763 , Ser473-Akt , and increased the nuclear translocation of phosphorylated P40763 time-dependently . Transfection of dominant negative Akt or application of PI3K inhibitorLY294002 significantly inhibited p-Tyr705- P40763 , p-Ser727- P40763 , and P49763 expression , indicating that Akt is involved in P52333 / P40763 / P49763 signaling cascade . To further examine whether P40763 binds to the promoter region of P49763 , Chip assay was used and it was found that P13725 could bind with P49763 promoter , which was inhibited by P52333 and PI3K inhibitors . Accumulation of P49763 in the pannus may contribute to the inflammation , angiogenesis and joints destruction in RA patients . These findings demonstrated the important role of P13725 in the pathology network of RA and provided novel therapeutic drug targets for RA treatment . 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 . 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 . 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 . Loss of androgen receptor expression promotes a stem-like cell phenotype in prostate cancer through P40763 signaling . P10275 ( AR ) signaling is important for prostate cancer progression . However , androgen-deprivation and/or AR targeting-based therapies often lead to resistance . Here , we demonstrate that loss of AR expression results in P40763 activation in prostate cancer cells . AR downregulation further leads to development of prostate cancer stem-like cells ( CSC ) , which requires P40763 . In human prostate tumor tissues , elevated cancer stem-like cell markers coincide with those cells exhibiting high P40763 activity and low AR expression . AR downregulation-induced P40763 activation is mediated through increased interleukin ( IL ) -6 expression . Treating mice with soluble P05231 receptor fusion protein or silencing P40763 in tumor cells significantly reduced prostate tumor growth and CSCs . Together , these findings indicate an opposing role of AR and P40763 in prostate CSC development .

MH_train_1146

MH_train_1146

MH_train_1146

interacts_with DB01045?

multiple_choice

Selective interaction of vitamin D receptor with transcriptional coactivators by a vitamin D analog . The nuclear vitamin D receptor ( P11473 ) is a member of a nuclear receptor superfamily and acts as a ligand-dependent transcription factor . A family of cotranscriptional activators ( Q15788 , Q15596 , and Q9Y6Q9 ) interacts with and activates the transactivation function of nuclear receptors in a ligand-dependent way . We examined interaction of P11473 with these coactivators that was induced by several vitamin D analogs , since they exert differential subsets of the biological action of vitamin D through unknown mechanisms . Unlike other vitamin D analogs tested , O75051 ( 22-oxa-1alpha,25-dihydroxyvitamin D3 ) induced interaction of P11473 with Q15596 but not with Q15788 or Q9Y6Q9 . Consistent with these interactions , only Q15596 was able to potentiate the transactivation function of P11473 bound to O75051 . Thus , the present findings suggest that the structure of P11473 is altered in a vitamin D analog-specific way , resulting in selective interactions of P11473 with coactivators . Such selective interaction of coactivators with P11473 may specify the array of biological actions of a vitamin D analog like O75051 , possibly through activating a particular set of target gene promoters . 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 . 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 . 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 . Characterization and functional analysis of cis-acting elements of the human farnesyl diphosphate synthetase ( P14324 ) gene 5' flanking region . Farnesyl diphosphate synthetase ( P14324 ) is a key enzyme in the isoprenoid pathway responsible for cholesterol biosynthesis , post-translational protein modifications and synthesis of steroid hormones , whose expression is regulated by phorbol esters and polyunsaturated fatty acids . Genomic comparison of the 5' upstream sequence of the P14324 genes identifies conserved binding sites for NF-Y , SP1 , SRE3 , and P25490 regulatory elements in rat , mouse , dog and chimpanzee . Two additional specific consensus sequences , upstream of the core promoter that had not been analysed previously , are shared only by human and chimpanzee genomes . The work presented here aimed at characterizing these genomic sequence elements in the human P14324 promoter region and their contribution to gene expression . We have characterized functionally the minimal basal promoter of the human P14324 gene by means of deletion mutants and we have identified two cis-acting elements which modulate the P14324 gene expression and are recognized by Pax5 and O75051 -1 transcription factors . 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 . 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 . 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 . 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 . 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 . Q07954 controls P47712 phosphorylation , O95477 expression and cellular cholesterol export . BACKGROUND : DB00171 -binding cassette transporter A1 mediates apolipoprotein AI-dependent efflux of cholesterol and thereby removes cholesterol from peripheral tissues . O95477 expression is tightly regulated and deficiency of this cholesterol transporter results in cholesterol accumulation within cells . P01130 -related protein 1 ( Q07954 ) participates in lipid metabolism and energy homeostasis by endocytosis of apolipoprotein E-containing lipoproteins and modulation of cellular proliferation signals . METHODS AND PRINCIPAL FINDINGS : In the present study , we demonstrate a new role for Q07954 in reverse cholesterol transport . Absence of Q07954 expression results in increased PDGFRbeta signaling and sequential activation of the mitogen-activated protein kinase signaling pathway , which increases phosphorylation of cytosolic phospholipase A(2) ( cPLA(2) ) . Phosphorylated and activated cPLA(2) releases arachidonic acid from the phospholipid pool . Overproduction of arachidonic acid suppresses the activation of LXR/RXR heterodimers bound to the promoter of LXR regulated genes such as O95477 , resulting in greatly reduced O95477 expression . CONCLUSIONS AND SIGNIFICANCE : Q07954 regulates LXR-mediated gene transcription and participates in reverse cholesterol transport by controlling cPLA(2) activation and O95477 expression . Q07954 thus functions as a physiological integrator of cellular lipid homeostasis with signals that regulate cellular proliferation and vascular wall integrity . Increased plasma levels of LDL cholesterol in rabbits after adenoviral overexpression of human scavenger receptor class B type I . Scavenger receptor class B type I ( Q8WTV0 ) , a P16671 family member , plays a key role in high-density lipoprotein ( HDL ) metabolism , reverse cholesterol transport , and whole body cholesterol homeostasis , and is shown to be involved in the development of atherosclerosis in mice . In this report , we describe the effects of the adenoviral overexpression of human Q8WTV0 ( hSR-BI ) in New Zealand White ( NZW ) rabbits , a wild-type animal model that expresses cholesteryl ester transfer protein ( P11597 ) in plasma , displays a manlike lipoprotein profile , and is susceptible to atherosclerosis . A total of 1x10(12) adenoviral particles containing either hSR-BI or lacZ complementary deoxyribonucleic acid ( control ) were infused into the ear vein of NZW rabbits . Transgene expression was ascertained by TaqMan Real Time polymerase chain reaction measurements . Rabbits infected with Ad/hSR-BI ( adenoviral plasmids containing hSR-BI ) showed a faster clearance of administered [3H]HDL cholesterol and significantly decreased apolipoprotein ( apo ) A-I levels when compared to control rabbits , respectively . Interestingly , we found markedly increased levels of low-density lipoprotein ( LDL ) cholesterol exclusively in Q8WTV0 -overexpressing rabbits . These changes were not accompanied by alterations in P01130 expression but by increased levels of CE transfer in these animals . By lowering HDL cholesterol and increasing plasma apoB-containing lipoprotein levels , the overexpression of Q8WTV0 leads to a lipoprotein pattern , which is believed to enhance the development of atherosclerosis . The role of Q8WTV0 in lipoprotein metabolism and atherogenesis in rabbits -- a P11597 -expressing animal model displaying a manlike lipoprotein profile -- may therefore be different from the one found in rodents . 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 . Maternal HLA- P06681 and 14 bp insertion in P17693 is associated with recurrent implantation failure after in vitro fertilization treatment . The major rate-limiting step in in vitro fertilization ( IVF ) success appears to be the implantation of the semi-allogeneic embryo into the maternal endometrium . To determine possible risk factors of recurrent failure of embryos to implant , we investigated immunogenetic determinants as level of human leukocyte antigen ( HLA ) histocompatibility , frequency of killer-cell immunoglobulin-like receptors ( P55040 ) and HLA-C alleles and P17693 polymorphism . We DNA typed women with recurrent implantation failure ( Q9HBH0 ) and their partners for classical HLA Class I , HLA Class II , P17693 and P55040 alleles and compared these results with couples with successful embryo implantation after their first IVF and normal fertile couples . No association was found between Q9HBH0 and the degree of histocompatibility between partners or sharing of a specific antigen . Also , no significant difference in P55040 haplotype or combination of HLA-C group and P55040 was observed . We did find a higher frequency of HLA- P06681 and a higher frequency of 14 base pair ( bp ) insertion in P17693 in women with Q9HBH0 . Therefore we conclude that the degree of histocompatibility between partners is not a determining factor for the occurrence of Q9HBH0 . However , presence of the HLA- P06681 allotype and the P17693 allele with a 14 bp insertion is a significant risk factor . 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 . 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 . Effects of pravastatin on the expression of DB00171 -binding cassette transporter A1 . In vitro inhibition of 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase causes the suppression of liver X receptor ( LXR ) activity . Because LXR regulates the expression of DB00171 -binding cassette transporter ( DB01048 ) A1 , which is involved in the high-density lipoprotein-related reverse cholesterol transport pathway , we examined the effects of an P04035 inhibitor pravastatin on O95477 expression in vitro and in vivo . DB00175 ( 10 microM ) significantly reduced the transcript levels of murine O95477 gene by 35 % in RAW264.7 macrophages under a lipoprotein-deficient condition . The inhibition was due to the decreased mevalonic acid production because addition of exogenous mevalonic acid restored O95477 mRNA levels . In addition , cholesterol and 22(R)-hydroxycholesterol thoroughly blunted the inhibition . Furthermore , pravastatin did not decrease O95477 mRNA and protein levels in HepG2 hepatocytes even in the absence of exogenous LXR agonists . Oral dosing of pravastatin ( 0.1 % concentration in drinking water ) for 24 h or 2 weeks to mice did not decrease O95477 mRNA and protein levels in the liver and leukocytes . Interestingly , pravastatin significantly increased both hepatic and leukocyte LXRalpha mRNA levels . Thus , although P04035 inhibitors suppress O95477 mRNA expression in the absence of LXR agonists , in vivo inhibition of P04035 is unlikely to cause such suppression . 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 . 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 . 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 . Blocking of P27361 and P28482 sensitizes human mesothelioma cells to doxorubicin . BACKGROUND : Malignant mesotheliomas ( MM ) have a poor prognosis , largely because of their chemoresistance to anti-cancer drugs such as doxorubicin ( Dox ) . Here we show using human MM lines that Dox activates extracellular signal-regulated kinases ( P27361 and 2 ) , causally linked to increased expression of ABC transporter genes , decreased accumulation of Dox , and enhanced MM growth . Using the Q02750 /2 inhibitor , U0126 and stably transfected shERK1 and shERK2 MM cell lines , we show that inhibition of both P27361 and 2 sensitizes MM cells to Dox . RESULTS : U0126 significantly modulated endogenous expression of several important drug resistance ( P10415 , P08183 , O15438 ) , prosurvival ( P10415 ) , DNA repair ( P38398 , P51587 ) , hormone receptor ( AR , Q92731 , PPARγ ) and drug metabolism ( P08684 ) genes newly identified in MM cells . In comparison to shControl lines , MM cell lines stably transfected with shERK1 or shERK2 exhibited significant increases in intracellular accumulation of Dox and decreases in cell viability . Affymetrix microarray analysis on stable shERK1 and shERK2 MM lines showed more than 2-fold inhibition ( p ≤ 0.05 ) of expression of DB00171 binding cassette genes ( P45844 , Q8WWZ7 , Q9BZC7 , MDR/TAP , O95477 , O94911 , Q92887 ) in comparison to shControl lines . Moreover , injection of human MM lines into SCID mice showed that stable shERK1 or shERK2 lines had significantly slower tumor growth rates in comparison to shControl lines after Dox treatment . CONCLUSIONS : These studies suggest that blocking P27361 and 2 , which play critical roles in multi-drug resistance and survival , may be beneficial in combination with chemotherapeutic drugs in the treatment of MMs and other 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 . Interaction of P42226 with its co-activator Q15788 / Q15788 is regulated by dephosphorylation of the latter via PP2A . Regulation of gene expression represents a central issue in signal-regulated cellular responses . P42226 is a critical mediator of P05112 stimulated gene activation . To mediate this function , P42226 recruits co-activator complexes . We have previously shown that P42226 binds the DB00233 -B domain of the co-activator Q15788 via an LXXLL motif in its transactivation domain . Our recent finding that the DB00233 -B domain of Q15788 is also essential for co-activator complex formation points to an additional level of regulation of the co-activator assembly . In this study , we discovered that dephosphorylation of Q15788 is essential for the interaction with P42226 and for P05112 -dependent transcriptional activation . PP2A dephosphorylates Q15788 and facilitates the activation of P42226 target genes . Interestingly , simultaneous inhibition of phosphatase and cyclin-dependent kinases rescues the Q15788 / P42226 interaction . Moreover , arrest of cells at P55008 /S results in enhanced Q15788 phosphorylation . In summary , our results indicate that the interaction of Q15788 and P42226 is dynamically regulated by the phosphatase PP2A and by cyclin-dependent kinases . This provides a mechanism for integrating transcriptional regulation by P42226 with cell cycle progression . Down-regulation of cholesterol 7alpha-hydroxylase ( P22680 ) gene expression by bile acids in primary rat hepatocytes is mediated by the c-Jun N-terminal kinase pathway . DB04540 7alpha-hydroxylase ( P22680 ) , the rate-limiting enzyme in the neutral pathway of bile acid biosynthesis , is feedback-inhibited at the transcriptional level by hydrophobic bile acids . Recent studies show that bile acids are physiological ligands for farnesoid X receptor ( Q96RI1 ) . Activated Q96RI1 indirectly represses P22680 transcription through induction of small heterodimer protein ( Q15466 -1 ) . In this study , we provide evidence that bile acids rapidly down-regulate P22680 transcription via activation of the JNK/c-Jun pathway . Furthermore , we demonstrate that Q15466 -1 is also a direct target of activated c-Jun . In primary rat hepatocyte cultures , taurocholate ( TCA ) strongly activated JNK in a time- and concentration-dependent manner . P01375 -alpha , a potent activator of JNK , also rapidly activated JNK and down-regulated P22680 mRNA levels . Overexpression of dominant-negative P45983 or a transactivating domain mutant of c-Jun significantly blocked the ability of TCA to down-regulate P22680 mRNA . In contrast , overexpression of wild-type c-Jun ( c-Jun(wt) ) enhanced the repression of P22680 by TCA . Moreover , overexpression of c-Jun(wt) resulted in increased Q15466 -1 promoter activity . Mutation of a putative AP-1 ( c-Jun ) element suppressed c-Jun-mediated activation of the Q15466 -1 promoter construct . These results indicate that the bile acid-activated JNK pathway plays a pivotal role in regulating P22680 levels in primary rat hepatocytes . Effect of lipopolysaccharide on the xenobiotic-induced expression and activity of hepatic cytochrome P450 in mice . Infection-associated inflammation can alter the expression levels and functions of cytochrome P450s ( CYPs ) . Cyp gene expression is regulated by the activation of several nuclear receptors , including pregnane X receptor ( O75469 ) , constitutive androstane receptor ( CAR ) , and aryl hydrocarbon receptor ( P35869 ) . These receptors can be activated by xenobiotics , including medicines . Here , to study the xenobiotic-induced fluctuations in CYP during inflammation , we examined the effect of lipopolysaccharide ( LPS ) treatment on the level of mRNAs encoding hepatic CYPs induced by xenobiotic-activated nuclear receptors , in mice . Both the mRNA induction of Cyp genes and the metabolic activities of CYP proteins were examined . LPS treatment caused a significant decrease in the induced expression of the mRNAs for Cyp3a11 , 2c29 , 2c55 , and 1a2 , but not for Cyp2b10 . To assess the CYP enzymatic activities , CYP3A-mediated testosterone 6β-hydroxylation and the intrinsic clearance ( CL(int) ) of nifedipine in liver microsomes were measured in mice treated with the xenobiotic pregnenolone-16alpha-carbonitrile ( Q15149 ) with or without LPS administration . Both assays revealed that the CYP3A activity , which was induced by Q15149 , declined significantly after LPS treatment , and this decline correlated with the Cyp3a11 mRNA level . In addition , we found that the mRNAs for interleukin ( IL ) -1β and tumor necrosis factor ( P01375 ) α were increased after treatment with LPS plus xenobiotics . Our findings demonstrated that LPS treatment reduces the O75469 - and P35869 -mediated , and possibly CAR-mediated Cyp gene expression and further suggest that these decreases are dependent on inflammatory cytokines in the liver . 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 . Genetic risk factors in recurrent venous thromboembolism : A multilocus , population-based , prospective approach . BACKGROUND : Recurrent venous thromboembolism ( VTE ) is a common , complex disorder ; however , genetic factors have been suggested to play a role in the disease development . We therefore conducted a multi-locus genetic study examining the potential associations of candidate gene variants in inflammation , thrombosis , coagulation , and lipid metabolism pathways , individually or interactively , with risk of recurrent VTE . METHODS : Using DNA samples collected at baseline in the Prevention of Recurrent Venous Thromboembolism trial ( PREVENT ) , we genotyped 86 candidate genes polymorphisms among 43 individuals who subsequently developed recurrent VTE and among 396 individuals who remained free of recurrent event over a mean follow-up period of 2.1 years to prospectively determine whether these gene polymorphisms contribute to the risk of recurrent VTE . RESULTS : Using a single-marker ' uncorrected ' analysis , P51681 A(-2459)G [ rs1799864 ] , P08254 5A(-1171)6A [ rs3025058 ] and P27169 gln192arg [ rs662 ] gene variants were associated with increased risk , and P11597 C(-629)A [ rs1800775 ] gene variant with reduced risk of recurrent VTE , respectively . Furthermore , potentially important gene-gene-interactions were detected by the Monte Carlo Markov chain Logic Regression method . CONCLUSIONS : Although the present findings are hypothesis-generating and require confirmation in an independent investigation , our study provides a practical example of detecting epistasis in common , complex diseases . 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 . Q9UEF7 ameliorates chemically induced endoplasmic reticulum ( ER ) stress signaling . BACKGROUND : Both endoplasmic reticulum ( ER ) stress , a fundamental cell response associated with stress-initiated unfolded protein response ( UPR ) , and loss of Q9UEF7 , an anti-aging hormone linked to NF-κB-induced inflammation , occur in chronic metabolic diseases such as obesity and type 2 diabetes . We investigated if the loss of Q9UEF7 is causally linked to increased ER stress . METHODS : We treated human renal epithelial HK-2 , alveolar epithelial A549 , HEK293 , and SH-SH-SY5Y neuroblastoma cells with ER stress-inducing agents , thapsigargin and/or tunicamycin . Effects of overexpression or siRNA-mediated knockdown of Q9UEF7 on UPR signaling was investigated by immunoblotting and Real-time PCR . RESULTS : Elevated Q9UEF7 levels in HK-2 cells decreased expression of ER stress markers phospho -- O75460 , XBP-1s , P11021 , P35638 , pJNK , and phospho-p38 , all of which were elevated in response to tunicamycin and/or thapsigargin . Similar results were observed using A549 cells for XBP-1s , P11021 , and P35638 in response to thapsigargin . Conversely , knockdown of Q9UEF7 in P29320 293 cells using siRNA caused further thapsigargin-induced increases in pIRE-1 , XBP-1s , and P11021 . Q9UEF7 overexpression in A549 cells blocked thapsigargin-induced caspase and PARP cleavage and improved cell viability . CONCLUSION : Our data indicate that Q9UEF7 has an important role in regulating ER stress and that loss of Q9UEF7 is causally linked to ER stress-induced apoptosis . 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 % . Activation by Q99572 agonists of two phospholipases A2 ( P04054 ) in ductal cells of rat submandibular gland . Coupling of the calcium-independent P04054 with kallikrein secretion . Isolated ductal cells of rat submandibular gland phospholipid pools were labeled with [3H]arachidonic acid ( AA ) . The tracer was incorporated preferentially to phosphatidylcholine ( 46 % of the lipidic fraction ) . Extracellular DB00171 induced the release of [3H]AA to the extracellular medium in a time- and dose-dependent manner ( EC50 = 220 microM ) . Among other agents tested , only 2 ' , 3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate ( Bz- DB00171 ) was able to mimic the effect of DB00171 ( EC50 = 15 microM ) , without activation of phospholipase C . The purinergic antagonists oxidized DB00171 , suramin , and Coomassie Blue partly inhibited the response to 1 mM DB00171 and 100 microM Bz- DB00171 ; the response was also blocked by the addition of Mg2+ or Ni2+ . Expression of Q99572 receptor mRNA in these cells was confirmed by reverse transcription-polymerase chain reaction . In the presence of extracellular calcium , the phospholipase A2 inhibitor 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid ( a nonspecific inhibitor ) , arachidonyl trifluoromethylketone ( AACOCF3 , an inhibitor of the calcium-dependent cytosolic P04054 ( P47712 ) ) , and bromoenol lactone ( an inhibitor of the calcium-independent P04054 ( iPLA2 ) ) inhibited the release of [3H]AA induced by DB00171 and Bz- DB00171 . In the absence of extracellular calcium , the release of [3H]AA in response to the purinergic agonists was still observed ; this response was not affected by AACOCF3 and completely blocked by bromoenol lactone . DB00171 and Bz- DB00171 stimulated a calcium-independent secretion of kallikrein , which could be blocked by BEL but which was enhanced by AACOCF3 . It is concluded that the Q99572 receptor in ductal cells is coupled to kallikrein secretion through a calcium-dependent P47712 and a calcium-independent iPLA2 . 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 . 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 . Microarray gene expression profiling analysis combined with bioinformatics in multiple sclerosis . Multiple sclerosis ( MS ) is the most prevalent demyelinating disease and the principal cause of neurological disability in young adults . Recent microarray gene expression profiling studies have identified several genetic variants contributing to the complex pathogenesis of MS , however , expressional and functional studies are still required to further understand its molecular mechanism . The present study aimed to analyze the molecular mechanism of MS using microarray analysis combined with bioinformatics techniques . We downloaded the gene expression profile of MS from Gene Expression Omnibus ( GEO ) and analysed the microarray data using the differentially coexpressed genes ( DCGs ) and links package in R and Database for Annotation , Visualization and Integrated Discovery . The regulatory impact factor ( Q9HBH0 ) algorithm was used to measure the impact factor of transcription factor . A total of 1,297 DCGs between MS patients and healthy controls were identified . Functional annotation indicated that these DCGs were associated with immune and neurological functions . Furthermore , the Q9HBH0 result suggested that Q13422 , BACH1 , P17676 , P18146 , P01100 may play central regulatory roles in controlling gene expression in the pathogenesis of MS . Our findings confirm the presence of multiple molecular alterations in MS and indicate the possibility for identifying prognostic factors associated with MS pathogenesis . 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 . Critical role of retinoid/rexinoid signaling in mediating transformation and therapeutic response of P52948 - P13631 leukemia . While the nucleoporin 98-retinoic acid receptor gamma ( P52948 - P13631 ) is the first P13631 fusion protein found in acute leukemia , its roles and the molecular basis in oncogenic transformation are currently unknown . Here , we showed that homodimeric P52948 - P13631 not only acquired unique nuclear localization pattern and ability of recruiting both P19793 and wild-type P52948 , but also exhibited similar transcriptional properties as P10276 fusions found in acute promyelocytic leukemia ( APL ) . Using murine bone marrow retroviral transduction/transformation assay , we further demonstrated that P52948 - P13631 fusion protein had gained transformation ability of primary hematopoietic stem/progenitor cells , which was critically dependent on the C-terminal GLFG domain of P52948 and the DNA binding domain ( DBD ) of P13631 . In contrast to other P52948 fusions , cells transformed by the P52948 - P13631 fusion were extremely sensitive to all-trans retinoic acid ( DB00755 ) treatment . Interestingly , while pan-RXR agonists , SR11237 and LGD1069 could specifically inhibit P52948 - P13631 transformed cells , mutation of the RXR interaction domain in P52948 - P13631 had little effect on its transformation , revealing that therapeutic functions of rexinoid can be independent of the direct biochemical interaction between RXR and the fusion . Together , these results indicate that deregulation of the retinoid/rexinoid signaling pathway has a major role and may represent a potential therapeutic target for P52948 - P13631 -mediated transformation . 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 . Impaired adipogenesis and lipolysis in the mouse upon selective ablation of the retinoid X receptor alpha mediated by a tamoxifen-inducible chimeric Cre recombinase ( Cre- P27361 ) in adipocytes . P19793 ( RXRalpha ) is involved in multiple signaling pathways , as a heterodimeric partner of several nuclear receptors . To investigate its function in energy homeostasis , we have selectively ablated the RXRalpha gene in adipocytes of 4-week-old transgenic mice by using the tamoxifen-inducible Cre- P27361 recombination system . Mice lacking RXRalpha in adipocytes were resistant to dietary and chemically induced obesity and impaired in fasting-induced lipolysis . Our results also indicate that RXRalpha is involved in adipocyte differentiation . Thus , our data demonstrate the feasibility of adipocyte-selective temporally controlled gene engineering and reveal a central role of RXRalpha in adipogenesis , probably as a heterodimeric partner for peroxisome proliferator-activated receptor gamma . Desmethyl DB00559 displays a similar in vitro interaction profile as DB00559 . The endothelin-1 receptor antagonists DB00559 and ambrisentan used for the treatment of pulmonary arterial hypertension remarkably differ in their potential to act as perpetrators in pharmacokinetic drug-drug interactions . So far , it is not clear whether the metabolites of DB00559 and ambrisentan contribute to the extent of drug interactions . We therefore investigated the effects of 4-hydroxymethyl ambrisentan , hydroxy DB00559 , desmethyl DB00559 , and hydroxy desmethyl DB00559 on targets which are inhibited or induced by the parent compounds . The hydroxylated metabolites of ambrisentan and DB00559 neither induced any of the genes investigated at the mRNA level , nor inhibited P-glycoprotein ( P-gp ) measured by calcein assay in L- P08183 cells , and only weakly inhibited organic anion transporting polypeptide ( P46721 ) 1B1 and Q9NPD5 measured by 8-fluorescein- DB02527 uptake in P29320 - Q9Y6L6 and P29320 - Q9NPD5 cells . In contrast , desmethyl DB00559 induced mRNA expression of cytochrome P450 3A4 ( P08684 , about 6-fold at 50 μM ) , P08183 ( P-gp , about 4.5-fold at 50 μM ) , and Q9UNQ0 ( breast cancer resistance protein , about 2-fold at 50 μM ) , whereas P33261 , O95342 , and Q92887 ( multidrug resistance-associated protein 2 ) were not induced in LS180 cells . In a reporter gene assay , desmethyl DB00559 activated pregnane X receptor with the highest potency of all metabolites tested . Whereas desmethyl DB00559 did not inhibit P-gp , it inhibited Q9Y6L6 with an IC50 of 3.8 μM ( 1.9-7.6 ) ( geometric mean , 95 % CI ) and Q9NPD5 with an IC50 of 7.4 μM ( 2.6-21.52 ) . In conclusion , our data demonstrate that desmethyl DB00559 exhibits a similar pharmacokinetic interaction profile as DB00559 and might contribute to the inducing effects of the parent compound . Pervanadate-induced shedding of the intercellular adhesion molecule ( ICAM ) -1 ectodomain is mediated by membrane type-1 matrix metalloproteinase ( P50281 ) . In several vascular diseases , the ectodomain of intercellular adhesion molecule ( ICAM ) -1 is shed by the proteolytic activity of a zinc-dependent endopeptidase , releasing a soluble form of the protein ( sICAM-1 ) , a common marker for inflammatory diseases . Since reactive oxygen species ( ROS ) generated during prolonged inflammation are known to induce shedding or cleavage of several transmembrane proteins , we sought to explore the cleavage and enzymatic effects that the pervanadate , via oxidation and subsequent inactivation of protein tyrosine phosphatase , has on P05362 cleavage . In these studies , we used endothelial cells ( ECs ) and 293 human embryonic kidney ( P29320 ) cells expressing high-levels of surface P05362 . In addition , use of specific tissue inhibitors of metalloproteinases ( TIMPs ) , small interfering (si)RNA designed to knockdown endopeptidase activity , and an immunocolocalization assay were employed to determine the identity of a specific metalloproteinase mediating pervanadate-induced sICAM-1 shedding . Our data indicate that membrane type-1 matrix metalloproteinase ( P50281 ) is involved in pervanadate-mediated shedding of the sICAM-1 ectodomain in both cell types . Immunostaining and confocal microscopy provide visual evidence that P05362 and P50281 colocalize at the cellular surface following pervanadate treatment , further implicating the involvement of P50281 activity in this mode of P05362 shedding . Self-assembly of P29320 cell-secreted ApoE particles resembles ApoE enrichment of lipoproteins as a ligand for the P01130 -related protein . Recent studies have shown that the lipidation and assembly state of apolipoprotein E ( apoE ) determine receptor recognition and amyloid-beta peptide ( Abeta ) binding . We previously demonstrated that apoE secreted by P29320 cells stably expressing apoE3 or apoE4 ( P29320 -apoE ) binds Abeta and inhibits Abeta-induced neurotoxicity by an isoform-specific process that requires apoE receptors . Here we characterized the structure of P29320 -apoE assemblies and determined their receptor binding specificity . By chromatography , P29320 -apoE elutes in high molecular mass fractions and is the size of plasma HDL , consistent with a multiprotein assembly . No lipid was associated with these apoE assemblies . Several methods for analyzing receptor binding indicate that P29320 -apoE is a ligand for low-density lipoprotein ( LDL ) receptor-related protein ( Q14764 ) but not the P01130 . This suggests that self-assembly of apoE may induce a functional conformation necessary for binding to Q14764 . Our results indicate that , in addition to lipid content , the assembly state of apoE influences Abeta binding and receptor recognition . Activation of the nuclear receptor O75469 decreases plasma LDL-cholesterol levels and induces hepatic steatosis in P01130 knockout mice . To investigate the potential for pregnane X receptor ( O75469 ) ligands as antiatherosclerotic drugs , we have determined the effect of O75469 activation on lipid metabolism in an established atherosclerotic mouse model . P01130 knockout mice were treated with the O75469 agonist Q15149 . Q15149 induced a striking 66 % decrease in plasma LDL-cholesterol levels . Q15149 did not affect the cholesterol levels of high-density lipoprotein ( HDL ) or very-low-density lipoprotein ( VLDL ) . VLDL-triglyceride levels were 2.2-fold increased by Q15149 , resulting in the presence of triglyceride-rich VLDL particles . This coincided with a 60 % decreased hepatic lipase ( HL ) -mediated plasma lipolysis rate , which could be attributed to a decrease in the hepatic mRNA expression level of both HL ( -31 % ) and its cofactor apolipoprotein A4 ( -62 % ) . In the liver , Q15149 induced a significant increase in the level of triglycerides ( +65 % ) and phospholipids ( +72 % ) , a hallmark of hepatic steatosis , leading to a marked increase in Oil red O neutral lipid staining . A similar effect was noticed in ApoE knockout mice . Our studies show that activation of the nuclear receptor O75469 by Q15149 leads to an inhibition of the plasma HL-mediated lipolysis rate , which is associated with a decrease in plasma LDL-cholesterol levels and induction of hepatic steatosis in P01130 knockout mice . 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 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 . 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 . 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 . 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 . 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 . miR-33 controls the expression of biliary transporters , and mediates statin- and diet-induced hepatotoxicity . Bile secretion is essential for whole body sterol homeostasis . Loss-of-function mutations in specific canalicular transporters in the hepatocyte disrupt bile flow and result in cholestasis . We show that two of these transporters , O95342 and O43520 , are functional targets of miR-33 , a micro-RNA that is expressed from within an intron of Q12772 . Consequently , manipulation of miR-33 levels in vivo with adenovirus or with antisense oligonucleotides results in changes in bile secretion and bile recovery from the gallbladder . Using radiolabelled cholesterol , we show that systemic silencing of miR-33 leads to increased sterols in bile and enhanced reverse cholesterol transport in vivo . Finally , we report that simvastatin causes , in a dose-dependent manner , profound hepatotoxicity and lethality in mice fed a lithogenic diet . These latter results are reminiscent of the recurrent cholestasis found in some patients prescribed statins . Importantly , pretreatment of mice with anti-miR-33 oligonucleotides rescues the hepatotoxic phenotype . Therefore , we conclude that miR-33 mediates some of the undesired , hepatotoxic effects of statins . 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 . 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 . High doses of atorvastatin and simvastatin induce key enzymes involved in VLDL production . Treatments with high doses of 3-hydroxy-3-methylglutaryl-coenzyme A ( HMG- DB01992 ) reductase inhibitors may induce the expression of sterol regulatory element binding protein ( SREBP ) -target genes , causing different effects from those attributed to the reduction of hepatic cholesterol content . The aim of this study was to investigate the effects of high doses of statins on the key enzymes involved in VLDL production in normolipidemic rats . To examine whether the effects caused by statin treatment are a consequence of P04035 inhibition , we tested the effect of atorvastatin on these enzymes in mevalonate-fed rats . DB01076 and simvastatin enhanced not only P04035 but also the expression of the Q12772 gene itself . As a result of the overexpression of Q12772 caused by the statin treatment , genes regulated basically by P36956 , as FA synthase and acetyl-coenzyme A carboxylase , were also induced and their mRNA levels increased . DAG acyltransferase and microsomal TG transfer protein mRNA levels as well as phosphatidate phosphohydrolase activity were increased by both statins . Simvastatin raised liver cholesterol content , ACAT mRNA levels , and P53007 :phosphocholine cytidylyltransferase activity , whereas it reduced liver DAG and phospholipid content . Mevalonate feeding reversed all changes induced by the atorvastatin treatment . These results show that treatment with high doses of statins induces key enzymes controlling rat liver lipid synthesis and VLDL assembly , probably as a result of Q12772 overexpression . Despite the induction of the key enzymes involved in VLDL production , both statins markedly reduced plasma TG levels , suggesting that different mechanisms may be involved in the hypotriglyceridemic effect of statins at high or low doses . 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 . Effects of APRIL ( O75888 ) polymorphisms and splicing isoforms on the secretion of soluble APRIL . Functional APRIL ( O75888 ) is a secreted trimer generated by furin protease cleavage . We previously reported the association of APRIL haplotypes formed by two nonsynonymous polymorphisms , Gly67Arg and Asn96Ser , with systemic lupus erythematosus . Here , we tested the hypothesis that polymorphisms and/or alternative splicing may influence the generation of soluble APRIL ( sAPRIL ) . P29320 293T cells were transfected with plasmids containing one of the six combinations of splicing isoforms ( α or β ) and haplotypes ( susceptible , neutral , or protective ) . APRIL concentrations were quantitated in the cell lysates and supernatants using an enzyme-linked immunosorbent assay ( ELISA ) . The association between splicing efficiency and polymorphisms was analyzed using quantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The efficiency of cleavage by furin protease was analyzed using western blotting . Although both splicing isoforms were cleaved by furin protease , sAPRIL was not detected in the supernatant of the cells transfected with the β isoform , regardless of the haplotype . This suggested that , similarly to B-cell activating factor ( Q9Y275 ) , one of the major APRIL splicing isoforms may not be secreted as a functional molecule . Furthermore , the secretion of sAPRIL was decreased in the transfectants expressing the protective haplotype . An association between the polymorphisms and splicing efficiency or furin cleavage efficiency was not detected . In conclusion , these observations suggested that both alternative splicing and polymorphisms may affect the generation of functional sAPRIL . 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 . Functional cooperation between Stat-1 and ets-1 to optimize icam-1 gene transcription . Intercellular adhesion molecule-1 ( P05362 ) plays an important role in the immune system , enabling the interactions between effector cells and target cells . It is also known to be involved in tumor growth and metastasis . Its expression is transcriptionally regulated by several proinflammatory cytokines including P01579 , which induces P05362 transcription via the JAK- P35610 signaling pathway in a Stat1-dependent fashion . The P05362 promoter contains several cis-active regulatory elements including 2 Ets binding sites ( EBSs ) located at positions -158 and -138 relatively to the AUG , which were previously shown to play a role in the constitutive activity of the P05362 promoter . In the present study , we have determined whether the EBSs are also involved in the regulation of P05362 gene transcription by pro-inflammatory cytokines . Transient transfection assays were performed with reporter genes containing P05362 promoter constructions cloned upstream from the firefly luciferase gene . Site-specific mutations of the Q9BTE0 diminished the promoter activity stimulated by P01579 , although the P01579 responsive element ( pIgammaRE ) , which binds Stat1 , was intact . Stimulation of the transcriptional activity following P01579 treatment was significantly reduced when both EBSs were inactivated . Co-immunoprecipitation experiments provided evidence of a physical interaction involving Ets1 and Stat1 . In COS-1 and P29320 293 cells cotransfected with P27918 -Stat1 and YFP-Ets fusion protein , fluorescence resonance energy transfer experiments confirmed the close proximity of these 2 proteins in living cells following treatment with P01579 . 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 . Comparative effects of cytokines on constitutive and inducible expression of the gene encoding for the cytochrome P450 3A6 isoenzyme in cultured rabbit hepatocytes : consequences on progesterone 6beta-hydroxylation . Cultured rabbit hepatocytes were used to compare the relative activities of cytokines to inhibit the constitutive or rifampicin ( Q9HBH0 ) -induced expression of the cytochrome P450 3A6 gene ( CYP3A6 ) . Human recombinant cytokines tested were interleukin-1beta ( IL-1beta ) ( 2 U/mL ) , interleukin-2 ( P60568 ) ( 5,000 U/mL ) and interferon-gamma ( P01579 ) ( 50 U/mL ) . Hepatocytes were cultured in the presence or absence of 25 microM Q9HBH0 for 24 hr , with or without cytokines alone or in combination . All these cytokines inhibited Q9HBH0 -induced P4503A6 expression without apparent cellular toxicity . By contrast , only P01579 treatment provided a significant decrease ( 41 % ) in the constitutive P4503A6 protein level . Moreover , cytokines differed in their ability to repress Q9HBH0 -dependent transcriptional induction of CYP3A6 : IL-1beta and P60568 were approximately equipotent , causing an almost 40-50 % suppression of CYP3A6 mRNA and protein levels , whereas P01579 exerted repressive effects only on P4503A6-related erythromycin N-demethylase activity and inducible protein expression . In fact , although strongly reducing P4503A6 protein content ( an approximate 70 % decrease ) , P01579 did not exhibit any influence on CYP3A6 mRNAs with the exception of its association with interleukins . All these results suggest that IL-1beta and P60568 mainly promote a transcriptional repression mechanism , given the absence of effect of these cytokines on the basal P4503A6 level , whereas P01579 exerts a post-transcriptional suppressive action on both induced and constitutive P4503A6 expression . Consequently , P4503A6-dependent progesterone 6beta-hydroxylase activity also presented a cytokine-specific pattern of inhibition , with a much greater sensitivity than P4503A6 immunoreactive protein to IL-1beta and P60568 + P01579 treatments . Thus , this study underlines the significant impact of inflammation on steroid metabolism . 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 . Superagonistic action of 14-epi-analogs of 1,25-dihydroxyvitamin D explained by vitamin D receptor-coactivator interaction . Two 14-epi-analogs of 1,25-dihydroxyvitamin D3 [ 1,25-(OH)(2)D(3) ] , 19-nor-14-epi-23-yne-1,25-(OH)2D3 ( TX522 ) and 19-nor-14,20-bisepi-23-yne-1,25-(OH)2D3 ( TX527 ) , show enhanced antiproliferative ( at least 10-fold ) and markedly lower calcemic effects both in vitro and in vivo , compared with 1,25-(OH)2D3 . This study aimed to evaluate their superagonistic effect at the level of interaction between the Vitamin D receptor ( P11473 ) and coactivators . Mammalian two-hybrid assays with VP16-fused P11473 and GAL4-DNA-binding-domain-fused steroid receptor coactivator 1 ( Q15788 ) , transcriptional intermediary factor 2 ( Tif2 ) , or Q15648 showed the 14-epi-analogs to be more potent inducers of P11473 -coactivator interactions than 1,25-(OH)2D3 ( up to 16- and 20-fold stronger induction of P11473 - Q15788 interaction for TX522 and TX527 at 10(-10) M ) . Similar assays in which metabolism of 1,25-(OH)2D3 was blocked with VID400 , a selective inhibitor of the 1,25-(OH)2D3-metabolizing enzyme Q07973 , showed that the enhanced potency of these analogs in establishing P11473 -coactivator interactions can only partially be accounted for by their increased resistance to metabolic degradation . Crystallization of TX522 complexed to the ligand binding domain of the human P11473 demonstrated that the epi-configuration of C14 caused the CD ring of the ligand to shift by 0.5 angstroms , thereby bringing the C12 atom into closer contact with Val300 . Moreover , C22 of TX522 made an additional contact with the CD1 atom of Ile268 because of the rigidity of the triple bond-containing side chain . The position and conformation of the activation helix H12 of P11473 was strictly maintained . In conclusion , this study provides deeper insight into the docking of TX522 in the P18428 and shows that stronger P11473 -coactivator interactions underlie the superagonistic activity of the two 14-epi-analogs . Activation of liver X receptor induces macrophage interleukin-5 expression . P05113 stimulates production of T15/EO6 IgM antibodies that can block the uptake of oxidized low density lipoprotein by macrophages , whereas a deficiency in macrophage P05113 expression accelerates development of atherosclerosis . Liver X receptors ( LXRs ) are ligand-activated transcription factors that can induce macrophage O95477 expression and cholesterol efflux , thereby inhibiting the development of atherosclerosis . However , it remains unknown whether additional mechanisms , such as the regulation of macrophage P05113 expression , are related to the anti-atherogenic properties of LXR . We initially defined P05113 expression in macrophages where the LXR ligand ( T0901317 ) induced macrophage P05113 protein expression and secretion . The overexpression of LXR increased , whereas its knockdown inhibited P05113 expression . Furthermore , we found that LXR activation increased P05113 transcripts , promoter activity , formation of an LXR·LXR-responsive element complex , and P05113 protein stability . In vivo , we found that T0901317 increased P05113 and total IgM levels in plasma and P05113 expression in multiple tissues in wild type mice . In P01130 knock-out ( P01130 (-/-) ) mice , T0901317 increased P05113 expression in the aortic root area . Taken together , our studies demonstrate that macrophage P05113 is a target gene for LXR activation , and the induction of macrophage P05113 expression can be related to LXR-inhibited 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 . 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 .

MH_train_1147

MH_train_1147

MH_train_1147

interacts_with DB00898?

multiple_choice

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 . Up-regulation of glutamate receptors in nucleus tractus solitarii underlies potentiation of baroreceptor reflex by heat shock protein 70 . Whereas induction of the 70-kDa heat shock protein ( HSP70 ) in the nucleus tractus solitarii ( P30990 ) , the terminal site in the brain stem for primary baroreceptor afferents , augments baroreceptor reflex ( BRR ) response , the underlying cellular and molecular mechanism is essentially unexplored . In Sprague-Dawley rats , we evaluated the hypothesis that HSP70 may potentiate BRR response by up-regulating the molecular synthesis and functional expression of glutamate receptors in the P30990 . Animals subjected to brief hyperthermic heat shock ( HS ; 42 degrees C for 15 min ) exhibited augmented expression of Q9UHB4 or Q12879 subunit of N-methyl-D-aspartate ( DB01221 ) receptors , GluR1 or P48058 subunits of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate receptors and Q16099 subunit of kainate receptors in the P30990 . Intriguingly , this up-regulation of glutamate receptors was preceded by an increase in HSP70 expression at the P30990 . The HS-induced augmentation in responsiveness of barosensitive P30990 neurons to transient hypertension or potentiation of BRR response was discernibly blunted by MK-801 or 6-cyano-7-nitroquinoxaline-2,3-dione . Bilateral microinjection into the P30990 of an antisense hsp70 oligonucleotide ( 50 pmol ) before HS significantly suppressed the induced expression of HSP70 or the increase in glutamate receptor subunits in the dorsal medulla and discernibly attenuated the potentiation of BRR response . Control microinjection into the P30990 of sense or scrambled hsp70 oligonucleotide ( 50 pmol ) was ineffective . These findings suggest that HSP70 induced by HS may enhance BRR response by up-regulating the molecular synthesis and functional expression of Q9UHB4 or Q12879 subunit of DB01221 receptors and GluR1 , P48058 , or Q16099 subunit of non- DB01221 receptors in the P30990 . 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 . 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 . 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 . Twenty bone-mineral-density loci identified by large-scale meta-analysis of genome-wide association studies . Bone mineral density ( BMD ) is a heritable complex trait used in the clinical diagnosis of osteoporosis and the assessment of fracture risk . We performed meta-analysis of five genome-wide association studies of femoral neck and lumbar spine BMD in 19,195 subjects of Northern European descent . We identified 20 BMD loci that reached genome-wide significance ( GWS ; P < 5 x 10(-8) ) , of which 13 map to regions not previously associated with this trait : 1p31.3 ( Q5T9L3 ) , 2p21 ( Q01082 ) , 3p22 ( P35222 ) , 4q21.1 ( Q9NQ76 ) , 5q14 ( Q06413 ) , 7p14 ( O95772 ) , 7q21.3 ( FLJ42280 ) , 11p11.2 ( O75096 , Q07960 , F2 ) , 11p14.1 ( Q6ZRR9 ) , 11p15 ( P35712 ) , 16q24 ( Q12952 ) , 17q21 ( Q9UQL6 ) and 17q12 ( P34998 ) . The meta-analysis also confirmed at GWS level seven known BMD loci on 1p36 ( Q9NUA8 ) , 6q25 ( P03372 ) , 8q24 ( O00300 ) , 11q13.4 ( O75197 ) , 12q13 ( Q8TDD2 ) , 13q14 ( O14788 ) and 18q21 ( Q9Y6Q6 ) . The many SNPs associated with BMD map to genes in signaling pathways with relevance to bone metabolism and highlight the complex genetic architecture that underlies osteoporosis and variation in BMD . Inhibitory effect of penta-acetyl geniposide on P13671 glioma cells metastasis by inhibiting matrix metalloproteinase-2 expression involved in both the PI3K and P29323 signaling pathways . Penta-acetyl geniposide [ ( Ac ) (5)GP ] , an acetylated geniposide product from Gardenia fructus , has been known to have hepatoprotective properties and recent studies have revealed its anti-proliferative and apoptotic effect on P13671 glioma cells . In this study , we first report the anti-metastastic effect of ( Ac ) (5)GP in the rat neuroblastoma line : P13671 glioma cells . First ( Ac ) (5)GP exhibited an inhibitory effect on abilities of adhesion and motility by cell-matrix adhesion assay , wound healing assay and Boyden chamber assay . Second , the decreasing activity of matrix metalloproteinase-2 ( P08253 ) was noted by gelatin zymography assay . Further analysis with semi-quantitative RT-PCR showed the mRNA levels of P08253 and membrane type I matrix metalloproteinase ( P50281 ) were significantly reduced , while the tissue inhibitor of matrix metalloproteinase-2 ( P16035 ) was elevated by ( Ac ) (5)GP treatment . Further ( Ac ) (5)GP also exerted an inhibitory effect on phosphoinositide 3-kinase ( PI3K ) protein expression , phosphorylation of extracellular signal-regulated kinases 1 and 2 ( P27361 /2 ) and inhibition of activation of transcription factor nuclear factor kappa B ( NF-kappaB ) , c-Fos , c-Jun . These findings proved ( Ac ) (5)GP is highly likely to be a inhibiting cancer migration agent to be further developed in the future . An P21860 / P04626 oncogenic unit plays a key role in Q02297 signaling and melanoma cell growth and survival . We recently identified neuregulin-1 ( Q02297 ) as a novel target of Notch1 required in Notch-dependent melanoma growth . P21860 and Q15303 , tyrosine kinase receptors specifically activated by Q02297 , have been shown to be either elevated in melanoma cell lines and tumors or to be mutated in 20 % of melanomas , respectively . While these data support key roles of Q02297 and its receptors in the pathogenesis of melanoma , whether P21860 and Q15303 display redundant or exclusive functions is not known . Here , we show that P21860 and Q15303 inhibition results in distinct outcomes . P21860 inhibition ablates the cellular responses to Q02297 , results in AKT inactivation and leads to cell growth arrest and apoptotic cell death . In contrast , Q15303 knockdown mildly affects cell growth , has no effects on cell survival and , importantly , does not alter the responses to Q02297 . Finally , we identified P04626 as a key coreceptor in Q02297 -dependent P21860 signaling . P04626 forms a complex with P21860 , and its inhibition recapitulates the phenotypes observed upon P21860 ablation . We propose that an Q02297 - P21860 - P04626 signaling unit operates in melanoma cells where it promotes growth and survival . Identification of cell surface targets for HIV-1 therapeutics using genetic screens . Human immunodeficiency virus ( HIV ) drugs designed to interfere with obligatory utilization of certain host cell factors by virus are less likely to encounter development of resistant strains than drugs directed against viral components . Several cellular genes required for productive infection by HIV were identified by the use of genetic suppressor element ( GSE ) technology as potential targets for anti-HIV drug development . Fragmented cDNA libraries from various pools of human peripheral blood mononuclear cells ( PBMC ) were expressed in vitro in human immunodeficiency virus type 1 ( HIV-1 ) -susceptible cell lines and subjected to genetic screens to identify GSEs that interfered with viral replication . After three rounds of selection , more than 15000 GSEs were sequenced , and the cognate genes were identified . The GSEs that inhibited the virus were derived from a diverse set of genes including cell surface receptors , cytokines , signaling proteins , transcription factors , as well as genes with unknown function . Approximately 2.5 % of the identified genes were previously shown to play a role in the HIV-1 life cycle ; this finding supports the biological relevance of the assay . GSEs were derived from the following 12 cell surface proteins : P61073 , CCR4 , P32248 , P20702 , P16070 , Q08722 , P34810 , Q07108 , P04233 , Q99062 , Q9UBS5 , and P20333 . Requirement of some of these genes for viral infection was also investigated by using RNA interference ( RNAi ) technology ; accordingly , 10 genes were implicated in early events of the viral life cycle , before viral DNA synthesis . Thus , these cell surface proteins represent novel targets for the development of therapeutics against HIV-1 infection and AIDS . 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 . 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 . 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 . Chemotherapeutic drugs and human tumor cells cytokine network . The ability of human tumor cell lines to produce various cytokines , chemokines , angiogenic and growth factors was investigated using Luminex multiplex technology . Media conditioned by tumor cells protected tumor cells from drug-induced apoptosis and stimulated tumor cell proliferation . Antibodies neutralizing P05231 , P10145 , P13500 and P13501 blocked this stimulation . Treatment of tumor cells with doxorubicin and cisplatin resulted in a substantial increase in the production of P05231 , P10145 , P13500 , P13501 , BFGF , G- P04141 and P15692 . This stimulation was associated with drug-induced activation of NF-kappaB , AP-1 , P05549 , CREB , Q9BYW2 , P35610 -1 , P35610 -3 , P35610 -5 and P39905 -2 transcription factors and upregulation of P05231 , P10145 , P09038 , P04141 -3 and P13501 gene expression . Treatment of tumor cells with doxorubicin and antibodies neutralizing DB00099 , P13500 or P13501 had higher inhibitory effects than each modality used alone . These results indicate that chemokines and growth factors produced by tumor by binding to the cognate receptors on tumor and stroma cells could provide proliferative and antiapoptotic signals helping tumor to escape drug-mediated destruction . Clinical studies showed that antibodies neutralizing P15692 ( DB00112 / DB00112 ) or blocking P04626 /neu signaling ( Herceptin/ DB00072 ) could increase the efficacy of chemotherapy , although these beneficial effects have been limited . It is possible that drug-stimulated production of growth and proangiogenic factors could counterbalance the effects of antibody therapy . In addition , numerous growth factors and chemokines share angiogenic and growth-stimulating properties , and thus reduction of a single factor is insufficient to completely block tumor growth . Thus , a broad disruption of tumor cytokine network is needed to further increase the efficacy of cancer therapy . 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 . 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 . 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 . 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 . DB00163 prevents ethanol-induced inflammatory , hormonal , and cytotoxic changes in reproductive tissues . DB00898 causes decreased function of the hypothalamic-pituitary-gonadal ( HPG ) axis . DB00898 resulted in inflammatory changes in HPG manifested by increased concentrations of pro-inflammatory cytokines . Since , such cytokines have deleterious effects on functions of HPG , it seemed possible that ethanol 's suppressive action could be due , at least in part , to this inflammation . Since oxidative stress can cause inflammation , we have used the antioxidant vitamin E to test , whether reducing inflammation might protect reproductive functions from ethanol . Rats were fed an ethanol diet or pair fed identically without ethanol for a 3-week period . For the last 10 days , animals were given 30 IU/kg or 90 IU/kg or vehicle . DB00898 significantly increased hypothalamic , pituitary and testicular P01375 and P05231 , all changes prevented by the higher dose of vitamin E. Also , ethanol induced changes in P01148 , LH , testosterone , and testicular germ cell apoptosis were similarly prevented by vitamin E . These data strikingly show that vitamin E protects the HPG from deleterious effects of ethanol and suggests that the mechanism of this protection might be both anti-inflammatory and antioxidant . Functional effect of polymorphisms in 15q25 locus on P30532 mRNA , bulky DNA adducts and P04637 mutations . Genome-wide association studies have demonstrated that genetic polymorphisms influence the risk of developing lung cancer . Nicotinic acetylcholine receptor alpha3 , alpha5 and beta4 genes ( P32297 , P30532 and P30926 ) cluster at the 15q25.1 lung cancer susceptibility locus . We genotyped 310 patients with non-small cell lung cancer and a control group of 348 cancer-free individuals for seven sequence variants located in P32297 and P30532 genes . Two of the polymorphisms ( rs3829787 and rs3841324 ) statistically influenced the risk of developing lung cancer . We found that four of the variants ( rs3829787 , rs3841324 , rs588765 and rs3743073 ) were associated with differential levels of genetic alterations measured as the levels of hydrophobic DNA adducts in the adjacent histologically normal tissue of the lung cancer patients and as P04637 mutations in their lung tumors . The seven sequence variants formed three haplotypes with a frequency above 5 % . The two most frequent haplotypes were associated with the risk of developing lung cancer and with smoking-related DNA alterations . We also found an association between P30532 mRNA levels and the sequence variants or haplotypes . In conclusion , our results showed that several of the polymorphisms and their haplotypes in P30532 / P32297 genes may have functional effects on ( i ) P30532 mRNA levels , ( ii ) polycyclic aromatic hydrocarbon-DNA adduct levels , ( iii ) P04637 mutations and ( iv ) susceptibility to lung cancer . 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 . 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 . P00750 has antiangiogenic properties without effect on tumor growth in a rat P13671 glioma model . P00750 ( tPA ) plays a major role in the fibrinolytic system . According to several reports , tPA may also have antiangiogenic properties , especially in combination with a free sulfhydryl donor ( FSD ) . In the rat P13671 glioma model , in vitro and in vivo tPA synthesis by glioma cells is enhanced by differentiation therapy . To address the antiangiogenic potential of tPA in this model , tPA was overexpressed in glioma tumors by ex vivo transduction of P13671 cells with a lentiviral vector encoding tPA . The transduced cells were subcutaneously implanted into nude mice . Gene transfer allowed for efficient synthesis of tPA by the P13671 tumors . Although the treatment of tPA+ tumor-bearing animals with the FSD captopril generated angiostatin in situ and reduced endothelial vascularization of the tumors , it had no effect on tumor growth . Alternative mechanisms could account for this lack of effect and consequently have important implications for vascular the treatment of glioblastoma . Innovative approaches for the development of antidepressant drugs : current and future strategies . Depression is a highly debilitating disorder that has been estimated to affect up to 21 % of the world population . Despite the advances in the treatment of depression with selective serotonin reuptake inhibitors ( SSRIs ) and serotonin and norepinephrine reuptake inhibitors ( SNRIs ) , there continue to be many unmet clinical needs with respect to both efficacy and side effects . These needs range from efficacy in treatment resistant patients , to improved onset , to reductions in side effects such as emesis or sexual dysfunction . To address these needs , there are numerous combination therapies and novel targets that have been identified that may demonstrate improvements in one or more areas . There is tremendous diversity in the types of targets and approaches being taken . At one end of a spectrum is combination therapies that maintain the benefits associated with SSRIs but attempt to either improve efficacy or reduce side effects by adding additional mechanisms ( P08908 , P28222 , P28221 , P28335 , alpha-2A ) . At the other end of the spectrum are more novel targets , such as neurotrophins ( P23560 , IGF ) , based on recent findings that antidepressants induce neurogenesis . In between , there are many approaches that range from directly targeting serotonin receptors ( P28335 , P50406 ) to targeting the multiplicity of potential mechanisms associated with excitatory ( glutamate , DB01221 , Q14416 , P41594 ) or inhibitory amino acid systems ( GABA ) or peptidergic systems ( neurokinin 1 , DB05394 1 , melanin-concentrating hormone 1 , V1b ) . The present review addresses the most exciting approaches and reviews the localization , neurochemical and behavioral data that provide the supporting rationale for each of these targets or target combinations . 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 . Altered gene expression in the spleen of adolescent rats following high ethanol concentration binge drinking . Binge drinking of alcoholic beverages among adolescents is a common practice that can have serious health consequences . DB00898 is a potent immunomodulator that alters a wide range of immune responses . However , it is unclear whether there is a differential immune response to alcoholic beverages with a high versus low concentration of ethanol . In this study , we used a PCR array containing 46 primer pairs of selected genes to compare mRNA expression in the spleen , an immune system organ , of adolescent rats following binge drinking of alcohol solutions containing either 20 % or 52 % ethanol ( v/v , 4.8 g/kg daily dosage ) , or water ( control ) for 3 d . We found that , expression of IL-1β , P05231 , P13500 , and GABA(A) receptor α2 subunit in the spleen were decreased , and P41594 and P46098 receptor expression were increased after administration of an ethanol solution containing 52 % ethanol , but not one with 20 % ethanol . Our data suggest that alcohol-mediated immunomodulatory effects are , in part , dependent on the ethanol by volume concentration . This is the first study to show that exposure to a high ethanol percentage beverage can have more profound effects on immune responses than one with a low percentage of ethanol . 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 . Central opioid inhibition of neuroendocrine stress responses in pregnancy in the rat is induced by the neurosteroid allopregnanolone . The hypothalamus-pituitary-adrenal ( Q9Y251 ) axis is the major neuroendocrine stress response system . P06850 ( P06850 ) neurons in the parvocellular paraventricular nucleus ( pPVN ) play a key role in coordinating responses of this system to stressors . The cytokine interleukin-1beta ( IL-1beta ) , mimicking infection , robustly activates these P06850 neurons via a noradrenergic input arising from the nucleus tractus solitarii ( P30990 ) . In late pregnancy , Q9Y251 axis responses to stressors , including IL-1beta , are attenuated by a central opioid mechanism that auto-inhibits noradrenaline release in the PVN . Here we show that the neuroactive progesterone metabolite allopregnanolone induces these changes in Q9Y251 responsiveness to IL-1beta in pregnancy . In late pregnancy , inhibition of 5alpha-reductase ( an allopregnanolone-synthesizing enzyme ) with finasteride restored Q9Y251 axis responses ( rapidly increased pPVN P06850 mRNA expression , DB01285 , and corticosterone secretion ) to IL-1beta . Conversely , allopregnanolone reduced Q9Y251 responses in virgin rats . In late pregnancy , activity of the allopregnanolone-synthesizing enzymes ( 5alpha-reductase and 3alpha-hydroxysteroid dehydrogenase ) was increased in the hypothalamus as was mRNA expression in the P30990 and PVN . Naloxone , an opioid antagonist , restores Q9Y251 axis responses to IL-1beta in pregnancy but had no additional effect after finasteride , indicating a causal connection between allopregnanolone and the endogenous opioid mechanism . Indeed , allopregnanolone induced opioid inhibition over Q9Y251 responses to IL-1beta in virgin rats . Furthermore , in virgin rats , allopregnanolone treatment increased , whereas in pregnant rats finasteride decreased proenkephalin-A mRNA expression in the P30990 . Thus , in pregnancy , allopregnanolone induces opioid inhibition over Q9Y251 axis responses to immune challenge . This novel opioid-mediated mechanism of allopregnanolone action may alter regulation of other brain systems in pregnancy . Fibroblast growth factor-2 in hyperplastic pituitaries of D2R knockout female mice . P14416 ( D2R ) knockout ( KO ) female mice develop chronic hyperprolactinemia and pituitary hyperplasia . Our objective was to study the expression of the mitogen fibroblast growth factor ( P09038 ) and its receptor , P11362 , comparatively in pituitaries from KO and wild-type ( WT ) female mice . We also evaluated P09038 subcellular localization and P09038 effects on pituitary function . P09038 -induced prolactin release showed a similar response pattern in both genotypes , even though basal and P09038 -stimulated release was higher in KO . P09038 stimulated pituitary cellular proliferation ( MTS assay and [(3)H]thymidine incorporation ) , with no differences between genotypes . P09038 concentration ( measured by ELISA ) in whole pituitaries or cultured cells was lower in KO ( P < 0.00001 and 0.00014 ) . Immunofluorescence histochemistry showed less P09038 in pituitaries from KO females and revealed a distinct P09038 localization pattern between genotypes , being predominantly nuclear in KO and cytosolic in WT pituitaries . Finally , P09038 could not be detected in the conditioned media from pituitary cultures of both genotypes . P11362 levels ( Western blot and immunohistochemistry ) were higher in pituitaries of KO . Basal concentration of phosphorylated ERKs was lower in KO cells ( P = 0.018 ) . However , when stimulated with P09038 , a significantly higher increment of P29323 phosphorylation was evidenced in KO cells ( P < or = 0.02 ) . We conclude that disruption of the D2R caused an overall decrease in pituitary P09038 levels , with an increased distribution in the nucleus , and increased P11362 levels . These results are important in the search for reliable prognostic indicators for patients with pituitary dopamine-resistant prolactinomas , which will make tumor-specific therapy possible . 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 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 . 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 . 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 . GABA Receptors Genes Polymorphisms and DB00898 Dependence : No Evidence of an Association in an Italian Male Population . OBJECTIVE : The genes encoding for gamma-aminobutyric acid ( GABA ) A and B receptors may be considered as candidates for alcoholism ; genetic alterations at this level may produce structural and functional diversity and thus play a role in the response to alcohol addiction treatment . To investigate these aspects further , we conducted a preliminary genetic association study on a population of Italian male alcohol addicts , focusing on GABA A and B receptors . METHODS : A total of 186 alcohol-dependent subjects ( in the first phase 139 , then 47 more samples ) and 182 controls were genotyped for 25 single nucleotide polymorphisms ( SNPs ) of genes encoding the alpha-1 subunit of GABA A receptor ( P14867 ) and subunits 1 and 2 of GABA B receptor ( Q9UBS5 and O75899 ) . The chi-squared test for allele and genotype distributions and Hardy-Weinberg equilibrium analysis of both subjects and controls were performed . Bonferroni 's correction for multiple comparisons was applied . RESULTS : Preliminary results comparing 139 alcohol-dependent subjects and 182 controls showed differences in genotype distribution in the former for SNP rs29253 , located in the intron region of the Q9UBS5 gene . In order to clarify the meaning of this association , 47 more samples from alcohol-dependent subjects were tested for this SNP only : the previously found association was not confirmed . CONCLUSION : The lack of significant differences between the two groups does not provide evidence that GABRA 1 and Q9UBS5 and 2 genes are candidates for alcoholism in this population . Further studies with larger samples are needed , together with investigation of other components of the GABA pathway . 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 . Nicotinic acetylcholine receptors containing the α6 subunit contribute to ethanol activation of ventral tegmental area dopaminergic neurons . DB00184 and alcohol are often co-abused suggesting a common mechanism of action may underlie their reinforcing properties . Both drugs acutely increase activity of ventral tegmental area ( VTA ) dopaminergic ( DAergic ) neurons , a phenomenon associated with reward behavior . Recent evidence indicates that nicotinic acetylcholine receptors ( nAChRs ) , ligand-gated cation channels activated by ACh and nicotine , may contribute to ethanol-mediated activation of VTA DAergic neurons although the nAChR subtype(s) involved has not been fully elucidated . Here we show that expression and activation of nAChRs containing the α6 subunit contribute to ethanol-induced activation of VTA DAergic neurons . In wild-type ( WT ) mouse midbrain sections that contain the VTA , ethanol ( 50 or 100 mM ) significantly increased firing frequency of DAergic neurons . In contrast , ethanol did not significantly increase activity of VTA DAergic neurons in mice that do not express Q15825 , the gene encoding the α6 nAChR subunit ( α6 knock-out ( KO ) mice ) . DB00898 -induced activity in WT slices was also reduced by pre-application of the α6 subtype-selective nAChR antagonist , α-conotoxin MII[E11A] . When co-applied , ethanol potentiated the response to ACh in WT DAergic neurons ; whereas co-application of ACh and ethanol failed to significantly increase activity of DAergic neurons in α6 KO slices . Finally , pre-application of α-conotoxin MII[E11A] in WT slices reduced ethanol potentiation of ACh responses . Together our data indicate that α6-subunit containing nAChRs may contribute to ethanol activation of VTA DAergic neurons . These receptors are predominantly expressed in DAergic neurons and known to be critical for nicotine reinforcement , providing a potential common therapeutic molecular target to reduce nicotine and alcohol co-abuse . 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 . P01375 alpha mediates GABA(A) receptor trafficking to the plasma membrane of spinal cord neurons in vivo . The proinflammatory cytokine TNFα contributes to cell death in central nervous system ( CNS ) disorders by altering synaptic neurotransmission . TNFα contributes to excitotoxicity by increasing P42262 -lacking AMPA receptor ( AMPAR ) trafficking to the neuronal plasma membrane . In vitro , increased AMPAR on the neuronal surface after TNFα exposure is associated with a rapid internalization of GABA(A) receptors ( GABA(A)Rs ) , suggesting complex timing and dose dependency of the CNS 's response to TNFα . However , the effect of TNFα on GABA(A)R trafficking in vivo remains unclear . We assessed the effect of TNFα nanoinjection on rapid GABA(A)R changes in rats ( N = 30 ) using subcellular fractionation , quantitative western blotting , and confocal microscopy . GABA(A)R protein levels in membrane fractions of TNFα and vehicle-treated subjects were not significantly different by Western Blot , yet high-resolution quantitative confocal imaging revealed that TNFα induces GABA(A)R trafficking to synapses in a dose-dependent manner by 60 min . TNFα-mediated GABA(A)R trafficking represents a novel target for CNS excitotoxicity . Chromosome 8p as a potential hub for developmental neuropsychiatric disorders : implications for schizophrenia , autism and cancer . Defects in genetic and developmental processes are thought to contribute susceptibility to autism and schizophrenia . Presumably , owing to etiological complexity identifying susceptibility genes and abnormalities in the development has been difficult . However , the importance of genes within chromosomal 8p region for neuropsychiatric disorders and cancer is well established . There are 484 annotated genes located on 8p ; many are most likely oncogenes and tumor-suppressor genes . Molecular genetics and developmental studies have identified 21 genes in this region ( ADRA1A , O15013 , Q15822 , Q15825 , Q05901 , Q9UBT3 , Q16555 , Q06889 , O60258 , Q9NP95 , P11362 , FZD3 , LDL , NAT2 , P07197 , Q02297 , PCM1 , P00750 , P48454 , Q8N474 and P54219 / P54219 ) that are most likely to contribute to neuropsychiatric disorders ( schizophrenia , autism , bipolar disorder and depression ) , neurodegenerative disorders ( Parkinson 's and Alzheimer 's disease ) and cancer . Furthermore , at least seven nonprotein-coding RNAs ( microRNAs ) are located at 8p . Structural variants on 8p , such as copy number variants , microdeletions or microduplications , might also contribute to autism , schizophrenia and other human diseases including cancer . In this review , we consider the current state of evidence from cytogenetic , linkage , association , gene expression and endophenotyping studies for the role of these 8p genes in neuropsychiatric disease . We also describe how a mutation in an 8p gene ( Fgf17 ) results in a mouse with deficits in specific components of social behavior and a reduction in its dorsomedial prefrontal cortex . We finish by discussing the biological connections of 8p with respect to neuropsychiatric disorders and cancer , despite the shortcomings of this evidence . 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 . 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 . 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 . Single-nucleotide polymorphisms of the dopamine D2 receptor increase inflammation and fibrosis in human renal proximal tubule cells . The dopamine D2 receptor ( D2R ) negatively regulates inflammation in mouse renal proximal tubule cells ( RPTCs ) , and lack or downregulation of the receptor in mice increases the vulnerability to renal inflammation independent of blood pressure . Some common single-nucleotide polymorphisms ( SNPs ; rs6276 , rs6277 , and rs1800497 ) in the human P14416 gene are associated with decreased D2R expression and function , as well as high blood pressure . We tested the hypothesis that human RPTCs ( hRPTCs ) expressing these SNPs have increased expression of inflammatory and injury markers . We studied immortalized hRPTCs carrying D2R SNPs and compared them with cells carrying no D2R SNPs . RPTCs with D2R SNPs had decreased D2R expression and function . The expressions of the proinflammatory tumor necrosis factor-α and the profibrotic transforming growth factor-β1 and its signaling targets P84022 and Snail1 were increased in hRPTC with D2R SNPs . These cells also showed induction of epithelial mesenchymal transition and production of extracellular matrix proteins , assessed by increased vimentin , fibronectin 1 , and collagen I a1 . To test the specificity of these D2R SNP effects , hRPTC with D2R SNPs were transfected with a plasmid encoding wild-type P14416 . The expression of D2R was increased and that of transforming growth factor-β1 , P84022 , Snail1 , vimentin , fibronectin 1 , and collagen I a1 was decreased in hRPTC with D2R SNPs transfected with wild-type P14416 compared with hRPTC-D2R SNP transfected with empty vector . These data support the hypothesis that D2R function has protective effects in hRPTCs and suggest that carriers of these SNPs may be prone to chronic renal disease and high blood pressure . 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 . T lymphocyte antigen 4-modified dendritic cell therapy for asthmatic mice guided by the P32248 chemokine receptor . The P33681 / P42081 - P10747 axis is a critical pathway for immuno-corrective therapy , and the cytotoxic T lymphocyte antigen 4 ( P16410 ) is a promising immunosuppressor targeting the P33681 / P42081 - P10747 axis ; however , its use for asthma therapy needs further optimization . A human P16410 fused with the IgCγ Fc ( DB01281 ) and mouse CC chemokine receptor type7 ( P32248 ) coding sequences were inserted into a recombinant adenovirus ( rAdV ) vector to generate rAdV- DB01281 and rAdV- P32248 . The naive dendritic cells ( DCs ) were infected with these rAdVs to ensure P32248 and DB01281 expression . The therapeutic effects of modified DCs were evaluated . rAdV- DB01281 and rAdV- P32248 infected DCs improved all asthma symptoms . Inflammatory cell infiltration and cytokine analysis showed that rAdV- DB01281 and rAdV- P32248 -modified DC therapy reduced the number of eosinophils and lymphocyte and neutrophil infiltration in the lung . Interestingly , assessment of the humoral immunity showed that the P05112 and IFNγ levels of the rAdV- DB01281 and rAdV- P32248 -modified DC-treated mice decreased significantly and did not reverse the Th1/Th2 balance . DCs expressing P32248 displayed guidance ability for DC migration , primarily for DCs in the inflammatory lung . Additionally , the rAdVs caused an inflammatory response by inducing DC differentiation , inflammatory cell infiltration and changes in cytokines ; however , mice transplanted with rAdV-green fluorescent protein ( GFP ) -infected DCs displayed no asthma manifestations . In conclusion , DB01281 -modified DCs exhibited a therapeutic effect on asthma , and P32248 may guide DC homing . The combination of these two molecules may be a model for precision-guided immunotherapy . Carbon-ion beam irradiation kills X-ray-resistant p53-null cancer cells by inducing mitotic catastrophe . BACKGROUND AND PURPOSE : To understand the mechanisms involved in the strong killing effect of carbon-ion beam irradiation on cancer cells with P04637 tumor suppressor gene deficiencies . MATERIALS AND METHODS : DNA damage responses after carbon-ion beam or X-ray irradiation in isogenic HCT116 colorectal cancer cell lines with and without P04637 ( p53+/+ and p53-/- , respectively ) were analyzed as follows : cell survival by clonogenic assay , cell death modes by morphologic observation of DAPI-stained nuclei , DNA double-strand breaks ( DSBs ) by immunostaining of phosphorylated P16104 ( γ P16104 ) , and cell cycle by flow cytometry and immunostaining of Ser10-phosphorylated histone H3 . RESULTS : The p53-/- cells were more resistant than the p53+/+ cells to X-ray irradiation , while the sensitivities of the p53+/+ and p53-/- cells to carbon-ion beam irradiation were comparable . X-ray and carbon-ion beam irradiations predominantly induced apoptosis of the p53+/+ cells but not the p53-/- cells . In the p53-/- cells , carbon-ion beam irradiation , but not X-ray irradiation , markedly induced mitotic catastrophe that was associated with premature mitotic entry with harboring long-retained DSBs at 24 h post-irradiation . CONCLUSIONS : Efficient induction of mitotic catastrophe in apoptosis-resistant p53-deficient cells implies a strong cancer cell-killing effect of carbon-ion beam irradiation that is independent of the p53 status , suggesting its biological advantage over X-ray treatment . Single-stranded DNA-binding proteins and neuron-restrictive silencer factor participate in cell-specific transcriptional control of the Q05586 gene . Our previous studies revealed that a proximal region of the N-methyl-D-aspartate receptor 1 ( Q05586 ) promoter is important for cell-type-specific expression . We have now explored the contributions of several regulatory elements to this specificity . Deletion of the neuron-restrictive silencer element partially relieved the suppression of promoter activity in P13671 glioma and HeLa cells . An overlapping G(C/G)G/tandem Sp1-containing region crucial for both basal and nerve growth factor ( P01138 ) -regulated promoter activity specifically bound nuclear proteins on its purine-rich sense strand . A faster migrating complex , single-stranded binding protein complex 1 ( SBPC1 ) , was highly enriched in HeLa cells , whereas a slower migrating complex , SBPC2 , was enriched in PC12 cells . A high ratio of 2/1 complex correlated with a high level of promoter activity . P01138 treatment of PC12 cells reduced SBPC1 but increased SBPC2 . Competition experiments showed that the SBPC1 binding required a dG4 sequence and the SBPC2 needed a core of TG3A plus a 5'-flanking sequence . Single-stranded DNA encompassing TG3A and/or dG4 specifically suppressed cotransfected Q05586 promoter activity . UV cross-linking studies indicated that a 31.5-kDa protein mainly formed SBPC1 , whereas SBPC2 contained several larger proteins . Our results suggest that neuron-restrictive silencer factor and single-stranded DNA-binding proteins may both play a role in cell-type specificity of the Q05586 gene , and the latter may also be involved in basal and P01138 -regulated activity . [ 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 . 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 . Low ethanol concentration alters P30532 RNA levels during early human development . DB00898 use is common and consumption during pregnancy has been shown to lead to a myriad of physical and neurologic abnormalities commonly referred to as fetal alcohol spectrum disorder . Substance addiction , which includes alcohol , has been shown to involve the major nicotinic acetylcholine receptor subunit P30532 . Using human embryonic stem cells as a model of early human development , we show that low concentrations of ethanol ( 20mM ) can alter the expression of P30532 . Changes in P30532 expression is linked to altered GABA and DB01221 receptor expression , as well as abnormal development of the frontal cortex . These results suggest that alcohol exposure can alter early neurologic development , which may favor addiction and other developmental abnormalities in unborn children . 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 . 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 . 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 . Predictive value of circulating interleukin-6 and heart-type fatty acid binding protein for three months clinical outcome in acute cerebral infarction : multiple blood markers profiling study . INTRODUCTION : There is no single blood marker for predicting the prognosis in ischemic stroke . A combination of multiple blood markers may enhance the ability to predict long-term outcome following ischemic stroke . METHODS : Blood concentrations of neuronal markers ( neuron-specific enolase , visinin-like protein 1 , heart type fatty acid binding protein ( hFABP ) and neuroglobin ) , astroglial markers ( P04271 and glial fibrillary acidic protein ) , inflammatory markers ( P05231 , P01375 -α , and P02741 ) , blood-brain barrier marker ( matrix metalloproteinase 9 ) , and haemostatic markers ( D-dimer and P05121 ) were measured within 24 hours after stroke onset . The discrimination and reclassification for favorable and poor outcome were compared after adding individual or a combination of blood markers to the clinical model of stroke outcome . RESULTS : In multivariate analysis , natural log-transformed ( log ) P05231 ( odds ratio ( OR ) : 1.75 , 95 % CI : 1.25 to 2.25 , P=0.001 ) and loghFABP ( OR : 3.23 , 95 % CI : 1.44 to 7.27 , P=0.005 ) were independently associated with poor outcome . The addition of a single blood marker to the clinical model did not improve the discriminating ability of the clinical model of stroke outcome . However , the addition of the combination of logIL-6 and loghFABP to the clinical model showed improved discrimination ( area under receiver operating characteristic ( AUROC ) curve : 0.939 versus 0.910 , P=0.03 ) and reclassification performance ( net reclassification improvement index : 0.18 , P=0.005 ) . CONCLUSIONS : A combination of circulating P05231 and hFABP level has an additive clinical value for the prediction of stroke outcome . 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 . 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 . 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 . 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 . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . 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 .

MH_train_1148

MH_train_1148

MH_train_1148

interacts_with DB08895?

multiple_choice

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 . Incorporating age at onset of smoking into genetic models for nicotine dependence : evidence for interaction with multiple genes . DB00184 dependence is moderately heritable , but identified genetic associations explain only modest portions of this heritability . We analyzed 3369 SNPs from 349 candidate genes and investigated whether incorporation of SNP-by-environment interaction into association analyses might bolster gene discovery efforts and prediction of nicotine dependence . Specifically , we incorporated the interaction between allele count and age at onset of regular smoking ( AOS ) into association analyses of nicotine dependence . Subjects were from the Collaborative Genetic Study of DB00184 Dependence and included 797 cases ascertained for Fagerström nicotine dependence and 811 non-nicotine-dependent smokers as controls , all of European descent . Compared with main effect models , SNP x AOS interaction models resulted in higher numbers of nominally significant tests , increased predictive utility at individual SNPs and higher predictive utility in a multi-locus model . Some SNPs previously documented in main effect analyses exhibited improved fits in the joint analysis , including rs16969968 from P30532 and rs2314379 from Q9Y6R4 . P30532 exhibited larger effects in later-onset smokers , in contrast with a previous report that suggested the opposite interaction ( Weiss et al. 2008 ) . However , a number of SNPs that did not emerge in main effect analyses were among the strongest findings in the interaction analyses . These include SNPs located in Q13224 ( P = 1.5 x 10(-5) ) , which encodes a subunit of the N-methyl-D-aspartate receptor channel , a key molecule in mediating age-dependent synaptic plasticity . Incorporation of logically chosen interaction parameters , such as AOS , into genetic models of substance use disorders may increase the degree of explained phenotypic variation and constitutes a promising avenue for gene discovery . 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 . 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 . 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 . 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 . 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 . JAK- P35610 and JAK-PI3K-mTORC1 pathways regulate telomerase transcriptionally and posttranslationally in ATL cells . Adult T-cell leukemia ( ATL ) is a heterogeneous tumor that is resistant to chemotherapy . Telomerase activity plays a critical role in tumorigenesis and is associated with the prognosis of ATL patients . Interleukin ( IL ) -2 commonly promotes tumor growth in chronic ATL cells . The signaling pathways involved in P60568 -regulated telomerase activation were studied in ATL cells derived from chronic ATL patients . P60568 challenge enhanced tyrosine phosphorylation of Janus-activated kinase (JAK)1-3 and P42229 , and induced P23458 and O60674 to associate with P42229 in P60568 -dependent ATL cells . Chromatin immunoprecipitation assays revealed that P42229 directly bound to the human telomerase reverse transcriptase ( hTERT ) promoter . P42229 short interfering RNA inhibited hTERT transcription in P60568 -stimulated ATL cells . Inhibitors of PI3K , HSP90 , and P42345 reduced P60568 -induced hTERT mRNA , protein expression , and telomerase activity . AKT , HSP90 , P42345 , S6 kinase , and hTERT immunoprecipitate from P60568 -stimulated cells contained telomerase activity , suggesting that hTERT directly interacts with , and is regulated by , these proteins . Binding of the p85 regulatory subunit of PI3K to O60674 was enhanced in an P60568 -dependent manner , indicating that O60674 propagates activation signals from the P60568 receptor and links hTERT activation to both the P42229 and PI3K pathways . Finally , P60568 -induced activation of telomerase and P42229 was observed in primary leukemic cells . These results indicate that P60568 stimulation induces hTERT activation through the JAK/ P35610 pathway and the JAK/PI3K/AKT/HSP90/mTORC1 pathway in P60568 -responsive ATL cells . These signaling proteins represent novel and promising molecular therapeutic targets for P60568 -dependent ATL . 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 . Mechanism of anti-inflammatory effect of tricin , a flavonoid isolated from Njavara rice bran in LPS induced hPBMCs and carrageenan induced rats . Njavara is an indigenous medicinal rice variety traditionally used in Ayurvedic system of medicine practiced in Kerala , India . Tricin is a bioflavonoid present in significantly higher levels in rice bran of Njavara . Present study attempted to identify the molecular target of tricin in TLR mediated signaling pathways by using lipopolysaccharide ( LPS ) induced human peripheral blood mononuclear cells ( hPBMCs ) and carrageenan induced paw edema in rats as experimental models . Tricin acted upstream in the activation of inflammation cascade by interfering with O00206 activation , preferably by blocking the LPS induced activation of O00206 , Q99836 and Q8IUC6 proteins in hPBMCs . Subsequently , tricin significantly blocked the activation of downstream kinases like p38MAPK , P45983 /2 and Q14653 . Thus the inhibitory effect of tricin on NF-κB and Q14653 together confirms the specific inhibition of both Q99836 dependent and Q8IUC6 dependent pathways . Tricin treatment also inhibited the pro-inflammatory effect of LPS by blocking the O00206 signaling mediated activation of cytosolic phospholipase A2 ( P47712 ) , which is confirmed by specific inhibition of P35354 . Results demonstrated that in addition to NF-κB , tricin can prevent the activation of P35610 proteins by significantly inhibiting the activation of both P42224 and P40763 via the down regulation of upstream phosphorylating enzymes like P23458 and O60674 . The protective anti-inflammatory effect of tricin was also confirmed by in vivo experiments . Thus , this study provides strong evidence that tricin exerts its anti-inflammatory effect via a mechanism involving the O00206 /NF-κB/ P35610 signaling cascade . 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 . Dysfunctional Immune-Mediated Inflammation in Rheumatoid Arthritis Dictates that Development of Anti-Rheumatic Disease Drugs Target Multiple Intracellular Signaling Pathways . A skewed repertoire of pro-inflammatory cytokines produced by the Th1 subset , one of the hallmarks of rheumatoid arthritis ( RA ) , is characterized by an overabundance of pro-inflammatory cytokines . P01375 -α , interleukin- 1 ( IL-1 ) , P05231 , P13232 , P10145 , Q9HBE4 , IL-12/IL-23 , P40933 , Q16552 , Q14116 , P24001 , and interferon-γ are primarily responsible for immune-mediated inflammation of RA by activating Janus kinases ( JAK ) -1 , -2 , -3 , p38 kinase , C-Jun-Nterminal kinase , extracellular signal-regulated kinase 1/2 and the phosphatidylinosotide-3-kinase/Akt/mTor pathways . Activation of these signaling pathways results in up-regulation of pro-inflammatory cytokines , cyclooxygenase-2 , matrix metalloproteinases , pro-angiogenesis proteins and anti-apoptosis proteins , the latter resulting in abnormal survival of activated T- and B-cells . Further , Q16552 also regulates the differentiation of P01730 + T-helper cells by inducing a Th17 T-cell subset , and a subpopulation of T-regulatory ( Treg ) cells . Although Treg cells are sufficiently abundant in RA synovial fluid , they fail to induce immune tolerance suggesting a functional deficiency likely coupled to putative protein kinase signaling abnormalities . The results of in vitro and studies in animal models of arthritis have indicated that inhibiting individual signaling pathways can blunt the synthesis of several of the pro-inflammatory biomarkers characteristic of human RA pathology . However , RA clinical trials indicated that small molecule inhibitors of P23458 , -2- , 3 and/or p38 kinase while exhibiting acceptable safety and tolerability profiles have only marginal and transient clinical effectiveness . These results suggested that future RA clinical studies using these or other kinase inhibitors will have to consider strategies designed to simultaneously inhibit multiple kinase pathways . Thyroid transcription factor-1 facilitates cerebrospinal fluid formation by regulating aquaporin-1 synthesis in the brain . In the brain , aquaporin-1 ( P29972 ) , a water channel for high osmotic water permeability , is mainly expressed in the apical membrane of the ventricular choroid plexus and regulates formation of cerebrospinal fluid ( P04141 ) . Although the physiology of P29972 has been the subject of several publications , much less is known about the trans-acting factors involved in the control of P29972 gene expression . Here we report that Q15669 -1 , a homeodomain-containing transcriptional regulator , is coexpressed with P29972 in the rat brain choroid plexus and enhances P29972 gene transcription by binding to conserved core Q15669 -1-binding motifs in the 5'-flanking region of the P29972 gene . Intracerebroventricular administration of an antisense Q15669 -1 oligodeoxynucleotide significantly decreased P29972 synthesis and reduced P04141 formation . In addition , blockade of Q15669 -1 synthesis increased survival of the animals following acute water intoxication-induced brain edema . These results suggest that Q15669 -1 is physiologically involved in the transcriptional control of P29972 , which is required for P04141 formation . 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 . 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 . Airway epithelium mediates the anti-inflammatory effects of exercise on asthma . Airway epithelium plays an important role in the asthma physiopathology . Aerobic exercise decreases Th2 response in murine models of allergic asthma , but its effects on the structure and activation of airway epithelium in asthma are unknown . BALB/c mice were divided into control , aerobic exercise , ovalbumin-sensitized and ovalbumin-sensitized plus aerobic exercise groups . Ovalbumin sensitization occurred on days 0 , 14 , 28 , 42 , and aerosol challenge from day 21 to day 50 . Aerobic exercise started on day 22 and ended on day 50 . Total cells and eosinophils were reduced in ovalbumin-sensitized group submitted to aerobic exercise . Aerobic exercise also reduced the oxidative and nitrosative stress and the epithelial expression of Th2 cytokines , chemokines , adhesion molecules , growth factors and NF-kB and Q99572 receptor . Additionally , aerobic exercise increased the epithelial expression of P22301 in non-sensitized and sensitized animals . These findings contribute to the understanding of the beneficial effects of aerobic exercise for chronic allergic airway inflammation , suggesting an immune-regulatory role of exercise on airway epithelium . 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 . [ 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 . DB11320 reduces susceptibility to natural killer cells via down-regulation of P26718 ligands on human monocytic leukaemia THP-1 cells . Natural killer ( NK ) group 2D ( P26718 ) is a key activating receptor expressed on NK cells , whose interaction with ligands on target cells plays an important role in tumorigenesis . However , the effect of histamine on P26718 ligands on tumour cells is unclear . Here we showed that human monocytic leukaemia THP-1 cells constitutively express MHC class I-related chain A ( Q29983 ) and Q9BZM6 on their surface , and incubation with histamine reduced the expression in a dose-dependent and time-dependent manner as assessed by flow cytometry . Interferon-γ augmented the surface expression of the P26718 ligands , and this augmentation was significantly attenuated by histamine . The histamine H1 receptor ( P35367 ) agonist 2-pyridylethylamine and P25021 agonist dimaprit down-regulated the expression of P26718 ligands , and activation of P35367 and P25021 signalling by A23187 and forskolin , respectively , had the same effect , indicating that the histamine-induced down-regulation of P26718 ligands is mediated by P35367 and P25021 . Quantitative reverse transcription-PCR showed that mRNA levels of the P26718 ligands and relevant microRNAs were not significantly changed by histamine . DB11320 down-regulated the surface expression of endoplasmic reticulum protein 5 , and inhibition of matrix metalloproteinases did not impair this down-regulation , indicating that proteolytic shedding was not involved . Instead , pharmacological inhibition of protein transport and proteasome abrogated it , and histamine enhanced ubiquitination of Q29983 . Furthermore , histamine treatment significantly reduced susceptibility to NK cell-mediated cytotoxicity . These results suggest that histamine down-regulates P26718 ligands through the activation of an P35367 - and P25021 -mediated ubiquitin-proteasome pathway and consequently reduces susceptibility to NK cells . A fusion cytokine coupling P04141 to P15248 induces heterologous receptor clustering and P42224 hyperactivation through O60674 promiscuity . Cytokine receptors are randomly distributed on the cell surface membrane and are activated upon binding of their extracellular ligands to mediate downstream cellular activities . We hypothesized that pharmaceutical clustering of ligand-bound , activated receptors may lead to heretofore unrealized gain-of-function with therapeutically desirable properties . We here describe an engineered bifunctional cytokine borne of the fusion of Granulocyte Macrophage Colony Stimulating Factor ( P04141 ) and P15248 ( P15248 ) ( hereafter GIFT9 fusokine ) and demonstrate that it chaperones co-clustering of the functionally unrelated P04141 receptor ( GMCSFR ) and P15248 receptor ( Q01113 ) on cell surface of target cells . We demonstrate that GIFT9 treatment of MC/9 cells leads to transhyperphosphorylation of Q01113 -associated P42224 by GMCSFR-associated O60674 . We also show that Q01113 -associated P23458 and P52333 augment phosphorylation of GMCSFR-linked P42229 . The functional relevance of these synergistic JAK/ P35610 transphosphorylation events translates to an increased mitogenic response by GMCSFR/ Q01113 -expressing primary marrow mast cells . The notion of inducing heterologous receptor clustering by engineered fusokines such as GIFT9 opens the door to a novel type of biopharmaceutical platform where designer fusokines modulate cell physiology through clustering of targeted receptor complexes . 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 . 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 . 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 . 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 . 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 . P60568 regulates expression of C- O75444 in human P01730 T cells . Blockade of IL-2R with humanized anti-CD25 Abs , such as daclizumab , inhibits Th2 responses in human T cells . Recent murine studies have shown that P60568 also plays a significant role in regulating Th2 cell differentiation by activated P42229 . To explore the role of activated P42229 in the Th2 differentiation of primary human T cells , we studied the mechanisms underlying P60568 regulation of C- O75444 expression . Chromatin immunoprecipitation studies revealed that P60568 induced P42229 binding to specific sites in the C- O75444 promoter . These sites corresponded to regions enriched for markers of chromatin architectural features in both resting P01730 and differentiated Th2 cells . Unlike P05231 , P60568 induced C- O75444 expression in P01730 T cells with or without prior TCR stimulation . TCR-induced C- O75444 expression was significantly inhibited by treatment with daclizumab or a P52333 inhibitor , R333 . Furthermore , P60568 and P05231 synergistically induced C- O75444 expression in TCR-activated T cells , suggesting functional cooperation between these cytokines . Finally , both TCR-induced early P05112 mRNA expression and P05112 cytokine expression in differentiated Th2 cells were significantly inhibited by IL-2R blockade . Thus , our findings demonstrate the importance of P60568 in Th2 differentiation in human T cells and support the notion that IL-2R-directed therapies may have utility in the treatment of allergic disorders . 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 . 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 . Activation by Q99572 agonists of two phospholipases A2 ( P04054 ) in ductal cells of rat submandibular gland . Coupling of the calcium-independent P04054 with kallikrein secretion . Isolated ductal cells of rat submandibular gland phospholipid pools were labeled with [3H]arachidonic acid ( AA ) . The tracer was incorporated preferentially to phosphatidylcholine ( 46 % of the lipidic fraction ) . Extracellular DB00171 induced the release of [3H]AA to the extracellular medium in a time- and dose-dependent manner ( EC50 = 220 microM ) . Among other agents tested , only 2 ' , 3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate ( Bz- DB00171 ) was able to mimic the effect of DB00171 ( EC50 = 15 microM ) , without activation of phospholipase C . The purinergic antagonists oxidized DB00171 , suramin , and Coomassie Blue partly inhibited the response to 1 mM DB00171 and 100 microM Bz- DB00171 ; the response was also blocked by the addition of Mg2+ or Ni2+ . Expression of Q99572 receptor mRNA in these cells was confirmed by reverse transcription-polymerase chain reaction . In the presence of extracellular calcium , the phospholipase A2 inhibitor 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid ( a nonspecific inhibitor ) , arachidonyl trifluoromethylketone ( AACOCF3 , an inhibitor of the calcium-dependent cytosolic P04054 ( P47712 ) ) , and bromoenol lactone ( an inhibitor of the calcium-independent P04054 ( iPLA2 ) ) inhibited the release of [3H]AA induced by DB00171 and Bz- DB00171 . In the absence of extracellular calcium , the release of [3H]AA in response to the purinergic agonists was still observed ; this response was not affected by AACOCF3 and completely blocked by bromoenol lactone . DB00171 and Bz- DB00171 stimulated a calcium-independent secretion of kallikrein , which could be blocked by BEL but which was enhanced by AACOCF3 . It is concluded that the Q99572 receptor in ductal cells is coupled to kallikrein secretion through a calcium-dependent P47712 and a calcium-independent iPLA2 . 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 . Targeting of active P42345 inhibits primary leukemia T cells and synergizes with cytotoxic drugs and signaling inhibitors . OBJECTIVE : Rationally designed therapies aim at the specific disruption of critical signaling pathways activated by malignant transformation or signals from the tumor microenvironment . Because mammalian target of rapamycin ( P42345 ) is an important signal integrator and a key translational regulator , we evaluated its potential involvement in T-cell acute lymphoblastic leukemia ( T-ALL ) and whether P42345 blockade synergizes with chemotherapeutic agents or other signaling antagonists to inhibit primary leukemia T cells . MATERIALS AND METHODS : P42345 signaling status was assessed using biochemical , immunostaining , and molecular regulation studies and functional assays performed to assess the impact of P42345 blockade on T-ALL proliferation , survival , and cell cycle . RESULTS : We observed that P42345 signaling is highly activated in all T-ALL patients tested , with phosphorylation of its downstream substrates eIF4G and S6 ribosomal protein . P42345 activation was detected in vivo and was further increased in vitro by stimulation with interleukin-7 , a potentially leukemogenic cytokine normally produced by the bone marrow microenvironment . In T-ALL cells , P42345 blockade was associated with accumulation of the cyclin-dependent kinase inhibitor p27(kip1) , which preferentially adopted a nuclear localization . Functional studies using rapamycin or CCI-779 showed a dominant inhibitory effect of P42345 blockade on interleukin-7-induced proliferation , survival , and cell-cycle progression of T-ALL cells . Furthermore , P42345 blockade markedly potentiated the antileukemia effects of dexamethasone and doxorubicin , and showed highly synergistic interactions in combination with specific inhibitors of phosphatidylinositol 3-kinase/Akt and P52333 signaling . CONCLUSIONS : This study shows activation of P42345 signaling in primary T-ALL cells evolving in the leukemic bone marrow , and supports the inclusion of P42345 antagonists in current therapeutic regimens for this cancer . 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 . 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 . 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 . 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 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 . Identification of immunogenic regions within the alternative reading frame protein of hepatitis C virus ( genotype 3 ) . Hepatitis C virus ( HCV ) encodes ten classic proteins as well as a newly discovered alternative reading frame protein ( ARFP ) whose synthesis originates from the core region by a +1 frameshift . ARFP is produced by all HCV genotypes , but its function remains unknown . Although the immunogenicity of genotype 1- and 2-derived ARFP in infected hosts has been reported , no information is available for genotype 3-encoded ARFP . HCV genotype 3 core/ARFP region was PCR amplified , cloned , and sequenced . Recombinant ARFP and peptides were employed in ELISAs with patient serum samples . The effect of peptides on peripheral blood mononucleocytes ( PBMCs ) was also studied . DNA cloning and sequencing of HCV genotype 3 strain ( PKHCV3 ) revealed it to encode 160 aa ARFP , which harbors a C-terminal extension of 36 aa . Serum from 74 of 88 patients ( 84 % ) contained rARFP-reactive antibodies . Peptide ELISAs showed that all regions of rARFP were immunogenic , with peptide P08709 ( DSLSPRRAGAKAGPGLSPGT ) being the most immunodominant . When incubated with PBMCs from HCV-infected individuals , P08709 stimulated the production of TNFα and P22301 . PKHCV3-derived ARFP encodes a 160 aa protein and antibodies against its entire length are found in 84 % of all genotype 3-infected subjects . Peptide ELISAs revealed P08709 to be highly immunogenic and capable of eliciting impressive T-cell responses . Cytokine-induced human P01579 -secreting effector-memory Th cells in chronic autoimmune inflammation . T-helper ( Th ) cells activated by cytokines in the absence of T-cell receptor ligation are suspected to participate in inflammatory processes by production of interferon-gamma ( P01579 ) . Still , the relevance of such a mechanism has not been addressed in humans . Here we demonstrate that a subset of human effector-memory Th cells expressing functional interleukin-12R ( IL-12R ) , IL-18Ralpha , and P51681 ex vivo can be induced to secrete P01579 by cytokines signaling via the IL-2R common gamma-chain in combination with IL-12 and Q14116 . Cytokine-driven P01579 production depends on P52333 - and p38 mitogen-activated kinase signals and is sensitive to suppression by CD25(++) regulatory T cells . Contrary to P01579 (+) Th cells induced upon antigen-specific stimulation , their cytokine-activated counterparts characteristically lack expression of costimulator 4-1BB ( Q07011 ) . Strikingly , the majority of Th cells infiltrating inflamed joints of rheumatoid arthritis patients is equipped with receptors prerequisite for cytokine-induced P01579 secretion . Among these cells , we detected a substantial fraction that secretes P01579 directly ex vivo but lacks 4-1BB expression , indicating that cytokine-induced P01579 (+) Th cells operate in autoimmune inflammation . Our data provide a rationale for how human effector-memory Thcells can participate in perpetuating inflammatory processes in autoimmunity even in the absence of T-cell receptor ligation . 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 . P05112 up-regulates histamine H1 receptors by activation of H1 receptor gene transcription . DB11320 plays an important role in allergy mainly through histamine H1 receptor ( P35367 ) . Recent studies showed that the P35367 level is elevated in allergic conditions , suggesting that this will make the allergic symptoms worse by intensifying P35367 -mediated processes . Some cytokines are also involved in allergy , and interleukin-4 ( P05112 ) has been implicated as an important mediator of allergic inflammation . It is noteworthy that the level of P05112 is elevated under allergic states . We tested whether P05112 has a role in up-regulating P35367 level by using the cultured human HeLa cell as a model system that expresses both P05112 receptor and P35367 . P05112 stimulation increased P35367 protein levels and P35367 mRNA levels . P05112 also increased P35367 promoter activity , but had no effect on P35367 mRNA stability , indicating that up-regulation of P35367 was due to an increase in P35367 mRNA synthesis . P05112 activated P42226 ( signal transducer and activator of transcription 6 ) in HeLa cells , and up-regulation of P35367 mRNA and activation of P42226 by P05112 were inhibited by a specific P52333 ( Janus-activated kinase 3 ) inhibitor . Stimulation with histamine also up-regulated P35367 mRNA , and co-stimulation with histamine and P05112 elevated P35367 mRNA level significantly higher than the stimulation with histamine or P05112 alone did . These results indicated that P05112 up-regulated P35367 mRNA level through increased transcription of P35367 gene via P52333 - P42226 pathway . The effects of histamine and P05112 were additive , suggesting that these allergic mediators will work together to up-regulate P35367 level , and thus make the allergic symptom worse by intensifying P35367 -mediated allergic processes . Changes in gene expression in atherosclerotic plaques analyzed using DNA array . A better understanding of atherogenesis at the level of gene expression could lead to the identification of new therapeutic strategies for vascular diseases . With DNA array technology , it is possible to identify multiple , simultaneous changes in gene expression in small tissue samples from atherosclerotic arteries . We analyzed gene expression in normal arteries and in immunohistologically characterized human advanced atherosclerotic lesions using an array of 18376 cDNA fragments . The array method was first validated by detecting a group of genes ( n=17 ) that were already known to be connected to atherogenesis . These genes included e.g. P02649 , P34810 , P01033 and phospholipase D . Next we detected 75 differentially expressed genes that were previously not connected to atherogenesis . A subgroup of genes involved in cell signaling and proliferation was selected for further analyzes with in situ hybridization and RT-PCR which confirmed array results by showing induction in advanced lesions of P23458 ( P23458 ) which is an important signaling molecule in activated macrophages ; P15692 receptor-2 which mediates angiogenic and vasculoprotective effects of P15692 ; and an unknown gene , which mapped on chromosome 19 . It is concluded that DNA array technology enables fast screening of gene expression in small samples of atherosclerotic lesions . The technique will be useful for the identification of new factors , such as P23458 and P15692 receptor-2 , which may play an important role in atherogenesis . 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 . 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 . A negative feedback loop mediated by P40763 limits human Th17 responses . The transcription factor P40763 is critically required for the differentiation of Th17 cells , a T cell subset involved in various chronic inflammatory diseases . In this article , we report that P40763 also drives a negative-feedback loop that limits the formation of Q16552 -producing T cells within a memory population . By activating human memory P01730 (+)CD45RO(+) T cells at a high density ( HiD ) or a low density ( LoD ) in the presence of the pro-Th17 cytokines IL-1β , IL-23 , and TGF-β , we observed that the numbers of Th17 cells were significantly higher under LoD conditions . Assessment of P40763 phosphorylation revealed a more rapid and stronger P40763 activation in HiD cells than in LoD cells . Transient inhibition of active P40763 in HiD cultures significantly enhanced Th17 cell numbers . Expression of the P40763 -regulated ectonucleotidase P49961 , which catalyzes DB00171 hydrolysis , was higher in HiD , than in LoD , cell cultures . Interestingly , inhibition of P49961 ectonucleotidase activity enhanced Th17 responses under HiD conditions . Conversely , blocking the DB00171 receptor Q99572 reduced Th17 responses in LoD cultures . These data suggest that P40763 negatively regulates Th17 cells by limiting the availability of DB00171 . This negative-feedback loop may provide a safety mechanism to limit tissue damage by Th17 cells during chronic inflammation . Furthermore , our results have relevance for the design of novel immunotherapeutics that target the P40763 -signaling pathway , because inhibition of this pathway may enhance , rather than suppress , memory Th17 responses . 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 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 . 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 . 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 . 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 . A novel P01282 signaling pathway in T cells DB02527 --> protein tyrosine phosphatase ( Q06124 ? ) --> O60674 / Q14765 --> Th1 differentiation . Vasoactive intestinal peptide ( P01282 ) is a potent anti-inflammatory agent . In addition to the deactivation of macrophages , dendritic cells , and microglia , P01282 shifts the Th1/Th2 balance , promoting the preferential differentiation and survival of Th2 cells , to the detriment of the proinflammatory Th1 effectors . Several mechanisms operate in the Th1/Th2 shift induced by P01282 . Here we report on a novel mechanism for the effect of P01282 on T cell differentiation , and show that P01282 inhibits Th1 differentiation by interfering directly with the IL-12Jak2/ Q14765 signaling pathway in T cells . The effect of P01282 is DB02527 -dependent , and appears to be mediated through the activation of protein tyrosine phosphatases ( PTP ) , with Q06124 as a potential target . The activation of PTPs represents a novel DB02527 -downstream target for the immunomodulatory effects of P01282 . Interleukin 18 induces angiogenesis in vitro and in vivo via Src and Jnk kinases . BACKGROUND : Interleukin 18 ( Q14116 ) is a novel mediator of angiogenesis in rheumatoid arthritis ( RA ) . OBJECTIVE : To examine the role of Q14116 in RA angiogenesis and the signalling mechanisms involved . METHODS : Human dermal microvascular endothelial cell ( HMVEC ) chemotaxis , capillary morphogenesis assays and Matrigel plug angiogenesis assays were performed in vivo using Q14116 with or without signalling inhibitors . A novel model of angiogenesis was devised using dye-tagged HMVECs to study their homing into RA and normal ( NL ) synovial tissues ( STs ) engrafted in severe combined immunodeficient ( SCID ) mice . RESULTS : Q14116 -mediated angiogenesis depended on Src and Jnk , as the inhibitors of Src and Jnk blocked Q14116 -induced HMVEC chemotaxis , tube formation and angiogenesis in Matrigel plugs . However , inhibitors of O60674 , p38 , MEK , phosphatidylinositol-3-kinase and neutralising antibodies to vascular endothelial growth factor or stromal derived factor-1α did not alter Q14116 -induced HMVEC migration . These results were confirmed with Jnk or Src sense or antisense oligodeoxynucleotides . Moreover , Q14116 induced phosphorylation of Src and Jnk in HMVECs . As proof of principle , Q14116 null mice had a significantly decreased angiogenesis compared with wild-type mice in Matrigel plug angiogenesis assays in vivo . Q14116 markedly enhanced mature HMVEC homing to human RA ST compared with NL ST in SCID mice , confirming the role of Q14116 -induced angiogenesis in RA ST in vivo . CONCLUSION : Targeting Q14116 or its signalling intermediates may prove to be a potentially novel therapeutic strategy for angiogenesis-dependent diseases , such as RA . 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 . 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 . 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 . 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 . 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 . The Inhibition of Spinal Astrocytic O60674 - P40763 Pathway Activation Correlates with the Analgesic Effects of Triptolide in the Rat Neuropathic Pain Model . Neuropathic pain ( NP ) is an intractable clinical problem without satisfactory treatments . However , certain natural products have been revealed as effective therapeutic agents for the management of pain states . In this study , we used the spinal nerve ligation ( Q16658 ) pain model to investigate the antinociceptive effect of triptolide ( P28907 ) , a major active component of the traditional Chinese herb Tripterygium wilfordii Hook F . Intrathecal P28907 inhibited the mechanical nociceptive response induced by Q16658 without interfering with motor performance . Additionally , the anti-nociceptive effect of P28907 was associated with the inhibition of the activation of spinal astrocytes . Furthermore , intrathecal administration of P28907 attenuated Q16658 -induced janus kinase ( JAK ) signal transducers and activators of transcription 3 ( P40763 ) signalling pathway activation and inhibited the upregulation of proinflammatory cytokines , such as interleukin-6 , interleukin-1 beta , and tumour necrosis factor-α , in dorsal horn astrocytes . Moreover , Q13224 -containing spinal N-methyl D-aspartate receptor ( NMDAR ) was subsequently inhibited . Above all , P28907 can alleviate Q16658 -induced NP via inhibiting the neuroinflammation in the spinal dorsal horn . The anti-inflammation effect of P28907 may be related with the suppression of spinal astrocytic JAK- P40763 activation . Our results suggest that P28907 may be a promising drug for the treatment of NP . 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 . Interleukin 23 regulates proliferation of lung cancer cells in a concentration-dependent way in association with the interleukin-23 receptor . A proinflammatory cytokine , interleukin 23 ( IL-23 ) , plays a role in tumor progression by inducing inflammation in the tumor microenvironment , although there is debate about its role in carcinogenesis . Direct effects of IL-23 on tumor cells have been reported rarely , and contradictory effects have been observed . Here , we studied such effects of IL-23 in lung cancer cells in vitro and in vivo and explored the underlying mechanism . We found Q5VWK5 expression in tissues from lung adenocarcinoma and small cell carcinoma but not in lung squamous cell carcinoma tissue . Interestingly , different concentrations of IL-23 had opposite effects in the same types of cells . We confirmed that the different effects could be explained by differences in binding to the Q5VWK5 ( subunits IL-23r and IL-12Rβ1 ) . Low concentrations of IL-23 promoted the proliferation of Q5VWK5 -positive A549 and P08709 -1 lung cancer cells by binding to IL-23r , whereas high concentrations of IL-23 inhibited proliferation of these cells by binding to both IL-23r and IL-12Rβ1 . In contrast , IL-23 had no effect on Q5VWK5 -negative SK-MES-1 cells . IL-23 regulated the growth of human lung cancer cells through its effects on P40763 expression and phosphorylation in a concentration-dependent way ; the Ki-67 gene was involved in these processes . Our findings demonstrate for the first time that IL-23 affects the proliferation of Q5VWK5 -positive lung cancer cells and that this effect is dependent on the IL-23 concentration . This can explain at least part of the inconsistent reports on the role of IL-23 in the progression of carcinogenesis . 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 . The water channel aquaporin 1 is a novel molecular target of polychlorinated biphenyls for in utero anomalies . Despite serious health risks in humans and wild life , the underlying mechanisms that explain the gene-environment effects of chemical toxicants are largely unknown . Polychlorinated biphenyls ( PCBs ) are one of the most ubiquitous environmental toxicants worldwide , with reported epidemiological evidence for reproductive and neurocognitive anomalies in humans . Here , we show that Aroclor 1254 , a mixture of structurally distinct PCBs , causes preterm birth in interleukin ( IL ) -10(-/-) mice at a dose that does not show any adverse effects in wild type mice , highlighting the significance of P22301 as an anti-toxicant cytokine . Aroclor 1254-treated P22301 (-/-) mice demonstrated increased amniotic fluid , intrauterine growth restriction , and reduced litter size with postnatal neuromotor defects . Further , our results identify aquaporin 1 ( P29972 ) , a potent effector of fluid volume regulation and angiogenic activity , as a novel placental target of PCBs . In vivo or in vitro exposure to Aroclor 1254 coupled with P22301 deficiency significantly reduced the protein content of P29972 . Reduced uterine P29972 levels were associated with defective spiral artery transformation . Importantly , recombinant P22301 reversed P11498 -induced in vivo and in vitro effects . These data demonstrate for the first time that the P22301 - P29972 axis is a novel regulator of P11498 -induced in utero effects . 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 .

MH_train_1149

MH_train_1149

MH_train_1149

interacts_with DB01045?

multiple_choice

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 . [ 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 . 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 . 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 . 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 . 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 . 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 . Glucocorticoids do not alter developmental expression of hippocampal or pituitary steroid receptor coactivator-1 and -2 in the late gestation fetal guinea pig . Steroid receptor coactivator ( P12931 ) proteins interact with glucocorticoid receptors in a ligand-dependent manner to enhance transcription . Although glucocorticoids are essential for normal brain maturation , little is known about the presence or regulation of P12931 proteins in the developing central nervous system . In the current study we demonstrated that Q15788 was highly expressed in the fetal limbic system ( hippocampal P07451 > P00915 /2 > P22748 > dentate gyrus ) at gestational d ( gd ) 40 ( term , approximately 70 d ) , whereas P12931 -2 was undetectable at all time points . Hippocampal Q15788 mRNA and protein expression were reduced in male and female fetuses with advancing gestation . In contrast , Q15788 mRNA levels increased significantly in the dentate gyrus near term . Repeated maternal injection ( 1 or 10 mg/kg on gd 40 , 41 , 50 , 51 , 60 , and 61 ) with synthetic glucocorticoid had no effect on fetal limbic Q15788 expression at gd 62 in either sex . Q15788 and P12931 -2 mRNA expression in the anterior pituitary did not change over the second half of gestation and was unaffected by prenatal exposure to synthetic glucocorticoid . In conclusion , Q15788 expression undergoes spatial , temporal , and region-specific regulation during development , and limbic and pituitary Q15788 and P12931 -2 are not regulated by glucocorticoids in late gestation . Developmental changes in limbic Q15788 expression probably have important consequences on steroid receptor signaling , which is known to be critical for brain maturation in late gestation . Lipoprotein candidate genes for multivariate factors of the insulin resistance syndrome : a sib-pair linkage analysis in women twins . The insulin resistance syndrome ( P41252 ) is characterized by a combination of interrelated coronary heart disease risk factors , including low high-density lipoprotein cholesterol ( HDLC ) levels , obesity and increases in triglyceride ( TG ) , systolic and diastolic blood pressure ( BP ) , small low-density lipoprotein particles ( LDL-size ) , and fasting and postload plasma insulin and glucose . Using factor analysis , we previously identified multivariate factors based on data from women participating in the Kaiser Permanente Women Twins Study : 1 ) Weight/Fat , 2 ) P01308 / DB09341 , 3 ) Lipids , and 4 ) BP . The purpose of this study is to evaluate evidence for genetic linkage between the multivariate factors and candidate genes . Quantitative sib-pair analysis based on the factor scores with markers for 9 candidate genes was carried out based on data from 126 pairs of dizygotic ( DZ ) women twins from the second exam of the Kaiser Permanente Women Twins study . Suggestive evidence for linkage was found for the Weight/fat factor and the Apo E gene ( p = 0.01 ) , and stronger evidence for linkage with the Lipid factor and the cholesterol ester transfer protein ( p = 0.002 ) gene . Therefore , the P11597 gene appears to influence covariation in LDL size , TG , and HDL , and may account for a portion of the well-established statistical and metabolic associations observed between these risk factors . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 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 lipopolysaccharide on the xenobiotic-induced expression and activity of hepatic cytochrome P450 in mice . Infection-associated inflammation can alter the expression levels and functions of cytochrome P450s ( CYPs ) . Cyp gene expression is regulated by the activation of several nuclear receptors , including pregnane X receptor ( O75469 ) , constitutive androstane receptor ( CAR ) , and aryl hydrocarbon receptor ( P35869 ) . These receptors can be activated by xenobiotics , including medicines . Here , to study the xenobiotic-induced fluctuations in CYP during inflammation , we examined the effect of lipopolysaccharide ( LPS ) treatment on the level of mRNAs encoding hepatic CYPs induced by xenobiotic-activated nuclear receptors , in mice . Both the mRNA induction of Cyp genes and the metabolic activities of CYP proteins were examined . LPS treatment caused a significant decrease in the induced expression of the mRNAs for Cyp3a11 , 2c29 , 2c55 , and 1a2 , but not for Cyp2b10 . To assess the CYP enzymatic activities , CYP3A-mediated testosterone 6β-hydroxylation and the intrinsic clearance ( CL(int) ) of nifedipine in liver microsomes were measured in mice treated with the xenobiotic pregnenolone-16alpha-carbonitrile ( Q15149 ) with or without LPS administration . Both assays revealed that the CYP3A activity , which was induced by Q15149 , declined significantly after LPS treatment , and this decline correlated with the Cyp3a11 mRNA level . In addition , we found that the mRNAs for interleukin ( IL ) -1β and tumor necrosis factor ( P01375 ) α were increased after treatment with LPS plus xenobiotics . Our findings demonstrated that LPS treatment reduces the O75469 - and P35869 -mediated , and possibly CAR-mediated Cyp gene expression and further suggest that these decreases are dependent on inflammatory cytokines in the liver . 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 . The genomic clone P08908 which resembles a beta-adrenergic receptor sequence encodes the P08908 receptor . The recent cloning of the complementary DNAs and/or genes for several receptors linked to guanine nucleotide regulatory proteins including the adrenergic receptors ( alpha 1 , alpha 2A , alpha 2B , beta 1 , beta 2 ) , several subtypes of the muscarinic cholinergic receptors , and the visual ' receptor ' rhodopsin has revealed considerable similarity in the primary structure of these proteins . In addition , all of these proteins contain seven putative transmembrane alpha-helices . We have previously described a genomic clone , P08908 , isolated by cross-hybridization at reduced stringency with a full length beta 2-adrenergic receptor probe . This clone contains an intronless gene which , because of its striking sequence resemblance to the adrenergic receptors , is presumed to encode a G-protein-coupled receptor . Previous attempts to identify this putative receptor by expression studies have failed . We now report that the protein product of the genomic clone , Q96NT5 , transiently expressed in monkey kidney cells has all the typical ligand-binding characteristics of the 5-hydroxytryptamine ( P08908 ) receptor . Physical and functional interaction between the dopamine transporter and the synaptic vesicle protein synaptogyrin-3 . Uptake through the dopamine transporter ( Q01959 ) represents the primary mechanism used to terminate dopaminergic transmission in brain . Although it is well known that dopamine ( DA ) taken up by the transporter is used to replenish synaptic vesicle stores for subsequent release , the molecular details of this mechanism are not completely understood . Here , we identified the synaptic vesicle protein synaptogyrin-3 as a Q01959 interacting protein using the split ubiquitin system . This interaction was confirmed through coimmunoprecipitation experiments using heterologous cell lines and mouse brain . Q01959 and synaptogyrin-3 colocalized at presynaptic terminals from mouse striatum . Using fluorescence resonance energy transfer microscopy , we show that both proteins interact in live neurons . Pull-down assays with Q86UG4 ( glutathione S-transferase ) proteins revealed that the cytoplasmic N termini of both Q01959 and synaptogyrin-3 are sufficient for this interaction . Furthermore , the N terminus of Q01959 is capable of binding purified synaptic vesicles from brain tissue . Functional assays revealed that synaptogyrin-3 expression correlated with Q01959 activity in PC12 and MN9D cells , but not in the non-neuronal P29320 -293 cells . These changes were not attributed to changes in transporter cell surface levels or to direct effect of the protein-protein interaction . Instead , the synaptogyrin-3 effect on Q01959 activity was abolished in the presence of the vesicular monoamine transporter-2 ( Q05940 ) inhibitor reserpine , suggesting a dependence on the vesicular DA storage system . Finally , we provide evidence for a biochemical complex involving Q01959 , synaptogyrin-3 , and Q05940 . Collectively , our data identify a novel interaction between Q01959 and synaptogyrin-3 and suggest a physical and functional link between Q01959 and the vesicular DA system . 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 . P49327 inhibition induces differential expression of genes involved in apoptosis and cell proliferation in ocular cancer cells . P49327 ( P49327 ) , a lipogenic multienzyme complex , is overexpressed in the ocular cancer , retinoblastoma , and is strongly correlated with tumor invasion . Dietary nutrients are reported to exert anticancer effects through inhibition of lipid metabolism . Differential gene expression in cultured retinoblastoma cells induced by cerulenin , a chemical inhibitor of P49327 , was evaluated by cDNA microarray analysis . Cerulenin treatment resulted in significant upregulation of cytochrome c ( P99999 ) by 1.2-fold , whereas S-phase kinase-associated protein-2 ( Q13309 ) , a negative regulator of cell cycle , and the lipid metabolic genes ( Q07869 , P19793 , and O00763 ) were significantly downregulated by -1.59- , -1.8- , -1.83- , and -1.5-fold , respectively , in comparison with untreated cancer cells . The expressions of key differentially expressed genes were confirmed by quantitative real-time PCR . The altered expression of genes involved in cell proliferation , cell signaling , apoptosis , and cell cycle , correlated with the anticancer effects of cerulenin . P49327 inhibition may thus be a potential strategy in retinoblastoma management . [ Clinical and genetic characteristics of long-livers in Moscow region ] . In Moscow region long-livers we have studied distribution of P06858 , P11597 , P02649 , F2 , P12259 , P08709 , F13 , P02675 , P17301 , P05106 , P05121 , P42898 , Q9UBK8 , HLA- Q8IUH3 , HLA-DQA1 , P01920 genes polymorphisms , associated with predisposition to age pathology . Long-livers are characterized by favorable course of cardiovascular diseases accompanied by certain genetic factors . We have established that genotype H-H- of P06858 , allele epsilon2 of P02649 , genotype CC of P42898 ( 677C > T ) , genotype TC of P05106 , genotype GA of P02675 , HLA- Q8IUH3 *11 positively correlate with longevity . DB00819 : future perspective in topical glaucoma therapeutics . Through this review it is contemplated that acetazolamide ( Q9Y6V0 ) , an age-old treatment for glaucoma with a myriad of side effects and inadequate topical effectiveness , may be formulated into a topically effective agent by utilizing various newer formulation approaches of ocular drug delivery . Even though it has a poor solubility and penetration power , various studies mentioned in the review indicate that it is possible to successfully formulate topically effective Q9Y6V0 by using : ( i ) high concentration of the drug , ( ii ) surfactant gel preparations of Q9Y6V0 , ( iii ) Q9Y6V0 loaded into liposomes , ( iv ) cyclodextrins to increase the solubility and hence bioavailability of Q9Y6V0 , and ( v ) viscolyzers and other polymers either alone or in combination with cyclodextrins . With the advent of newer topical carbonic anhydrase inhibitors ( CAIs ) like dorzolamide and brinzolamide , a localized effect with fewer side effects is expected . But whenever absorbed systemically , a similar range of adverse effects ( attributable to sulphonamides ) may occur upon use . Furthermore , oral Q9Y6V0 is reported to be more physiologically effective than 2 % dorzolamide hydrochloride administered topically , even though in isolated tissues dorzolamide appears to be the most active as it shows the lowest IC(50) values for P00918 and P22748 [ M.F. Surgue , J. Ocular Pharmacol. Ther. 12 ( 1996 ) 363-376 ] . Hence , there exists considerable scope for the development of more/equally effective and inexpensive topically effective formulations of Q9Y6V0 . The use of various formulation technologies discussed in this review can provide a fresh impetus to research in this area . 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 . 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 . 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 . 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 . 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 . 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 % . 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 . 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 . 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 . 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 . Enhanced cellular responses and distinct gene profiles in human fetoplacental artery endothelial cells under chronic low oxygen . Fetoplacental endothelial cells are exposed to oxygen levels ranging from 2 % to 8 % in vivo . However , little is known regarding endothelial function within this range of oxygen because most laboratories use ambient air ( 21 % O2 ) as a standard culture condition ( SCN ) . We asked whether human umbilical artery endothelial cells ( HUAECs ) that were steadily exposed to the physiological chronic normoxia ( Q15149 , 3 % O2 ) for ∼20-25 days differed in their proliferative and migratory responses to P09038 and P15692 as well as in their global gene expression compared with those in the SCN . We observed that Q15149 enhanced P09038 - and P15692 -stimulated cell proliferation and migration . In oxygen reversal experiments ( i.e. , when Q15149 cells were exposed to SCN for 24 h and vice versa ) , we found that preexposure to 21 % O2 decreased the migratory ability , but not the proliferative ability , of the Q15149 -HUAECs in response to P09038 and P15692 . These Q15149 -enhanced cellular responses were associated with increased protein levels of Q16665 and NOS3 , but not P11362 , P17948 , and P35968 . Microarray analysis demonstrated that Q15149 up-regulated 74 genes and down-regulated 86 , 14 of which were directly regulated by hypoxia-inducible factors as evaluated using in silico analysis . Gene function analysis further indicated that the Q15149 -regulated genes were highly related to cell proliferation and migration , consistent with the results from our functional assays . Given that Q15149 significantly alters cellular responses to P09038 and P15692 as well as transcription in HUAECs , it is likely that we may need to reexamine the current cellular and molecular mechanisms controlling fetoplacental endothelial functions , which were largely derived from endothelial models established under ambient O2 . 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 . 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 . Pharmacogenomics and drug response in cardiovascular disorders . There are a total of 17 families of drugs that are used for treating the heterogeneous group of cardiovascular diseases . We propose a comprehensive pharmacogenomic approach in the field of cardiovascular therapy that considers the five following sources of variability : the genetics of pharmacokinetics , the genetics of pharmacodynamics ( drug targets ) , genetics linked to a defined pathology and its corresponding drug therapies , the genetics of physiologic regulation , and environmental-genetic interactions . Examples of the genetics of pharmacokinetics are presented for phase I ( cytochromes P450 ) and phase II ( conjugating enzymes ) drug-metabolizing enzymes and for phase III drug transporters . The example used to explain the genetics of pharmacodynamics is glycoprotein IIIa and the response to antiplatelet effects of aspirin . Genetics linked to a defined pathology and its corresponding drug therapies is exemplified by P08588 , P12821 , P11597 and P02649 and drug response in metabolic syndrome . The examples of cytochrome P450s , P02649 and P07550 in relation to ethnicity , age and gender are presented to describe genetics of physiologic regulation . Finally , environmental-genetic interactions are exemplified by P22680 and the effects of diet on plasma lipid levels , and by P02649 and the effects of smoking in cardiovascular disease . We illustrate this five-tiered approach using examples of cardiovascular drugs in relation to genetic polymorphism . 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 . 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 . 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 . Single nucleotide polymorphisms in P11597 , Q96NT5 , P41440 , P16671 , Q9HAY6 , Q6Q788 , and O95477 are significant predictors of plasma HDL in healthy adults . BACKGROUND : In a marker-trait association study we estimated the statistical significance of 65 single nucleotide polymorphisms ( SNP ) in 23 candidate genes on HDL levels of two independent Caucasian populations . Each population consisted of men and women and their HDL levels were adjusted for gender and body weight . We used a linear regression model . Selected genes corresponded to folate metabolism , vitamins B-12 , A , and E , and cholesterol pathways or lipid metabolism . METHODS : Extracted DNA from both the Sacramento and Beltsville populations was analyzed using an allele discrimination assay with a MALDI-TOF mass spectrometry platform . The adjusted phenotype , y , was HDL levels adjusted for gender and body weight only statistical analyses were performed using the genotype association and regression modules from the SNP Variation Suite v7 . RESULTS : Statistically significant SNP ( where P values were adjusted for false discovery rate ) included : P11597 ( rs7499892 and rs5882 ) ; Q96NT5 ( rs37514694 ; rs739439 ) ; P41440 ( rs3788199 ) ; P16671 ( rs3211956 ) ; Q9HAY6 ( rs6564851 ) , Q6Q788 ( rs662799 ) , and O95477 ( rs4149267 ) . Many prior association trends of the SNP with HDL were replicated in our cross-validation study . Significantly , the association of SNP in folate transporters ( Q96NT5 rs37514694 and rs739439 ; P41440 rs3788199 ) with HDL was identified in our study . CONCLUSIONS : Given recent literature on the role of niacin in the biogenesis of HDL , focus on status and metabolism of B-vitamins and metabolites of eccentric cleavage of β-carotene with lipid metabolism is exciting for future study . 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 . 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 . Global assessment of genetic variation influencing response to retinoid chemoprevention in head and neck cancer patients . Head and neck squamous cell carcinoma ( HNSCC ) patients are at an increased risk of developing a second primary tumor ( P21549 ) or recurrence following curative treatment . 13-cis-retinoic acid ( 13-cRA ) has been tested in chemoprevention clinical trials , but the results have been inconclusive . We genotyped 9,465 single nucleotide polymorphisms ( SNP ) in 450 patients from the Retinoid Head and Neck Second Primary Trial . SNPs were analyzed for associations with P21549 /recurrence in patients receiving placebo to identify prognosis markers and further analyzed for effects of 13-cRA in patients with these prognostic loci . Thirteen loci identified a majority subgroup of patients at a high risk of P21549 /recurrence and in whom 13-cRA was protective . Patients carrying the common genotype of rs3118570 in the retinoid X receptor ( P19793 ) were at a 3.33-fold increased risk ( 95 % CI , 1.67-6.67 ) and represented more than 70 % of the study population . This locus also identified individuals who received benefit from chemoprevention with a 38 % reduced risk ( 95 % CI , 0.43-0.90 ) . Analyses of cumulative effect and potential gene-gene interactions also implicated P30307 :rs6596428 and O60674 :rs1887427 as 2 other genetic loci with major roles in prognosis and 13-cRA response . Patients with all 3 common genotypes had a 76 % reduction in P21549 /recurrence ( 95 % CI , 0.093-0.64 ) following 13-cRA chemoprevention . Carriers of these common genotypes constituted a substantial percentage of the study population , indicating that a pharmacogenetic approach could help select patients for 13-cRA chemoprevention . The lack of any alternatives for reducing risk in these patients highlights the need for future clinical trials to prospectively validate our findings . TGFbeta1 , TNFalpha , and insulin signaling crosstalk in regulation of the rat cholesterol 7alpha-hydroxylase gene expression . The TGFbeta1/Smad pathway plays a critical role in cholestasis and liver fibrosis . Previous studies show that TGFbeta1 , TNFalpha , and insulin inhibit cholesterol 7alpha-hydroxylase ( P22680 ) gene transcription and bile acid synthesis in human hepatocytes . In this study , we investigated insulin , TGFbeta1 , and TNFalpha regulation of rat Cyp7a1 gene transcription . In contrast to inhibition of human P22680 gene transcription , TGFbeta1 stimulates rat Cyp7a1 reporter activity . P84022 , FoxO1 , and HNF4alpha synergistically stimulated rat Cyp7a1 gene transcription . Mutations of the P84022 , FoxO1 , or HNF4alpha binding site attenuated the rat Cyp7a1 promoter activity . Furthermore , TNFalpha and cJun attenuated TGFbeta1 stimulation of rat Cyp7a1 . P01308 or adenovirus-mediated expression of constitutively active P31749 inhibited FoxO1 and P84022 synergy . In streptozotocin-induced diabetic rats , Cyp7a1 mRNA expression levels were induced and insulin attenuated P22680 mRNA levels . Chromatin immunoprecipitation assay showed that FoxO1 binding to Cyp7a1 chromatin was increased in diabetic rat livers and insulin reduced FoxO1 binding . These results suggest a mechanistic basis for induction of Cyp7a1 activity and bile acid synthesis in cholestatic rats and in diabetic rats . The crosstalk of insulin , TGFbeta and TNFalpha signaling pathways may regulate bile acid synthesis and lipid homeostasis in diabetes , fatty liver disease , and liver fibrosis . Regulation of alpha-synuclein expression by liver X receptor ligands in vitro . P37840 is a lipid-binding protein expressed in neurons and oligodendrocytes which is increased in Parkinson 's disease . We identified two putative liver X receptor ( LXR ) response elements in the human alpha-synuclein gene and used synthetic ( TO901317 , GW3695 ) and physiological ( 27-hydroxycholesterol ) LXR activators to assess regulation of alpha-synuclein . LXR ligands upregulated alpha-synuclein mRNA by two-five-fold in human SK-N-SH neurons and three-six-fold in human Q03405 .13 oligodendrocytes . Significant 50 % to four-fold induction of alpha-synuclein protein was also detected . Under these conditions , mRNA for LXR-responsive gene O95477 was significantly upregulated 15-40-fold and 5-25-fold in neurons and oligodendrocytes , respectively . LXR may , therefore , contribute to the regulation of alpha-synuclein expression in neurons and oligodendrocytes . 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 . Gene expression analyses in cynomolgus monkeys provides mechanistic insight into high-density lipoprotein-cholesterol reduction by androgens in primates . Androgens increase muscle mass , decrease fat mass , and reduce high-density lipoprotein cholesterol ( HDL ) , but the relationship between body composition , lipoprotein metabolism , and androgens has not been explained . Here we treated ovariectomized cynomolgus monkeys with 5alpha-dihydrotestosterone ( DB02901 ) or vehicle for 14 d and measured lipoprotein and triglycerides . Nuclear magnetic resonance analysis revealed that DB02901 dose-dependently reduced the cholesterol content of large HDL particles and decreased mean HDL particle size ( P < 0.01 ) and also tended to lower low-density lipoprotein cholesterol without altering other lipoprotein particles . Liver and visceral fat biopsies taken before and after DB02901 treatment for 1 or 14 d were analyzed by genome-wide microarrays . In liver , DB02901 did not alter the expression of most genes involved in cholesterol synthesis or uptake but rapidly increased small heterodimer partner ( Q15466 ) RNA , along with concomitant repression of P22680 , a target of Q15466 transcriptional repression and the rate-limiting enzyme in bile acid synthesis . DB02901 regulation of Q15466 and P22680 also occurs in rats , indicating a conserved mechanism . In adipose tissue , pathway analyses suggested coordinate regulation of adipogenesis , tissue remodeling , and lipid homeostasis . Genes encoding P05019 and beta-catenin were induced , as were extracellular matrix , cell adhesion , and cytoskeletal components , whereas there was consistent down-regulation of genes involved in triacylglycerol metabolism . Interestingly , cholesterol ester transfer protein RNA was induced rapidly in monkey adipose tissue , whereas its inhibitor apolipoprotein CI was repressed . These data provide insight into the androgenic regulation of lipoprotein homeostasis and suggest that changes in adipose lipoprotein metabolism could contribute to HDL cholesterol reduction . Nuclear hormone receptors and cholesterol trafficking : the orphans find a new home . There are many homeostatic mechanisms that contribute to the regulation of cellular and serum cholesterol levels in humans . Much of our understanding of this regulation arose from studies of the cellular pathways controlling cholesterol synthesis and the uptake and degradation of low-density lipoproteins . The physiology governing cholesterol disposition in whole animals , including the molecular mechanisms responsible for dietary uptake of cholesterol and its subsequent catabolism to bile acids , are only now being uncovered . This review summarizes recent studies that have used modern genetic techniques , as well as cell biological methods , to begin to elucidate the pathways responsible for cholesterol trafficking in vivo . This work has led to the realization that networks of nuclear hormone receptors , including the LXR , Q96RI1 , Q07869 , and RXR proteins , play critical roles in lipid metabolism by virtue of their transcriptional regulation of the genes that control sterol metabolic pathways . Some of the major downstream targets of these regulatory networks involve members of the ABC transporter family , including O95477 , P45844 , Q9H222 , Q9H221 , P21439 /Mdr2 , and SPGP/ O95342 . These transporters contribute to the movement of sterols and biliary lipids across cellular membranes via mechanisms that have yet to be elucidated . The potential for manipulating these cholesterol trafficking pathways via drugs targeted to the nuclear hormone receptors has generated great interest in their biology and will undoubtedly lead to new therapeutic approaches to human disorders in the coming years . Differential gene expression in adipose tissue from obese human subjects during weight loss and weight maintenance . BACKGROUND : Differential gene expression in adipose tissue during diet-induced weight loss followed by a weight stability period is poorly characterized . Markers of these processes may provide a deeper understanding of underlying mechanisms . OBJECTIVE : We aimed to identify differentially expressed genes in human adipose tissue during weight loss and weight maintenance after weight loss . DESIGN : RNA from subcutaneous abdominal adipose tissue from 9 obese subjects was analyzed by using a complementary DNA microarray at baseline after weight loss on a low-calorie diet and after weight maintenance . RESULTS : Subjects lost 18.8 ± 1.8 % of weight and maintained this loss during weight maintenance ( 1.1 ± 2.1 % ; range : -9.3 to 10.6 % ) . Most differentially expressed genes exhibited a reciprocal regulation and returned to baseline after weight loss ( 2163 genes ) and weight maintenance ( 3175 genes ) . P11597 and P45844 , both of which participate in the HDL-mediated reverse cholesterol transport ( RCT ) , were among the most upregulated of the 750 genes that were differentially expressed after both processes . Several genes involved in inflammation were downregulated . The use of real-time polymerase chain reaction confirmed or partially confirmed the previously implicated genes Q9H2S6 and P14780 ( both downregulated ) , Q9NST1 ( upregulated ) , and O60543 and O00767 ( both reciprocally regulated ) . CONCLUSIONS : The beneficial effects of weight loss should be investigated after long-term weight maintenance . The processes of weight loss and weight maintenance should be viewed as biologically distinct . P11597 and P45844 may be important mediators of these effects through HDL-mediated RCT . Blood gene expression profiling of breast cancer survivors experiencing fibrosis . PURPOSE : To extend knowledge on the mechanisms and pathways involved in maintenance of radiation-induced fibrosis ( Q9HBH0 ) by performing gene expression profiling of whole blood from breast cancer ( BC ) survivors with and without fibrosis 3-7 years after end of radiotherapy treatment . METHODS AND MATERIALS : Gene expression profiles from blood were obtained for 254 BC survivors derived from a cohort of survivors , treated with adjuvant radiotherapy for breast cancer 3-7 years earlier . Analyses of transcriptional differences in blood gene expression between BC survivors with fibrosis ( n=31 ) and BC survivors without fibrosis ( n=223 ) were performed using R version 2.8.0 and tools from the Bioconductor project . Gene sets extracted through a literature search on fibrosis and breast cancer were subsequently used in gene set enrichment analysis . RESULTS : Substantial differences in blood gene expression between BC survivors with and without fibrosis were observed , and 87 differentially expressed genes were identified through linear analysis . Transforming growth factor-β1 signaling was identified as the most significant gene set , showing a down-regulation of most of the core genes , together with up-regulation of a transcriptional activator of the inhibitor of fibrinolysis , P05121 in the BC survivors with fibrosis . CONCLUSION : Transforming growth factor-β1 signaling was found down-regulated during the maintenance phase of fibrosis as opposed to the up-regulation reported during the early , initiating phase of fibrosis . Hence , once the fibrotic tissue has developed , the maintenance phase might rather involve a deregulation of fibrinolysis and altered degradation of extracellular matrix components . 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 . Peritoneal macrophages mediated delivery of chitosan/siRNA nanoparticle to the lesion site in a murine radiation-induced fibrosis model . BACKGROUND : Radiation-induced fibrosis ( Q9HBH0 ) is a dose-limiting complication of cancer radiotherapy and causes serious problems , i.e. restricted tissue flexibility , pain , ulceration or necrosis . Recently , we have successfully treated Q9HBH0 in a mouse model by intraperitoneal administration of chitosan/siRNA nanoparticles directed towards silencing P01375 alpha in local macrophage populations , but the mechanism for the therapeutic effect at the lesion site remains unclear . METHODS : Using the same murine Q9HBH0 model we utilized an optical imaging technique and fluorescence microscopy to investigate the uptake of chitosan/fluorescently labeled siRNA nanoparticles by peritoneal macrophages and their subsequent migration to the inflamed tissue in the Q9HBH0 model . RESULTS : We observed strong accumulation of the fluorescent signal in the lesion site of the irradiated leg up to 24 hours using the optical imaging system . We further confirm by immunohistochemical staining that Cy3 labeled siRNA resides in macrophages of the irradiated leg . CONCLUSION : We provide a proof-of-concept for host macrophage trafficking towards the inflamed region in a murine Q9HBH0 model , which thereby suggests that the chitosan/siRNA nanoparticle may constitute a general treatment for inflammatory diseases using the natural homing potential of macrophages to inflammatory sites . DB00171 -binding cassette transporter A1 R219K polymorphism and ischemic stroke risk in the Chinese population : a meta-analysis . Recently , many studies have been focused on the association between the DB00171 -binding cassette transporter A1 ( O95477 ) gene R219K polymorphism and ischemic stroke ( IS ) . However , the study results have been inconsistent , especially in the Chinese population . Therefore , we performed a meta-analysis to better clarify the association between the O95477 gene and IS . All of the relevant studies used in our meta-analysis were identified using PubMed , OVID , Cochrane Library , Chinese Wan Fang database , Chinese P01282 database , China National Knowledge Infrastructure ( CNKI ) , and China Biological Medicine Database ( CBM ) up to May 2013 . Statistical analysis was conducted with STATA software version 11.0 . Odds ratios with 95 % confidence intervals were applied to evaluate the strength of the association between O95477 gene R219K polymorphism and IS . Heterogeneity was evaluated using the Q-test and I(2) statistic . The funnel plots , Begg 's and Egger 's regression tests were used to assess the publication bias . Our meta-analysis showed the dominant genetic model ( OR=0.92 , 95 % CI : 0.88-0.96 ) , the recessive genetic model ( OR=0.73 , 95 % CI : 0.51-1.05 ) , the homozygote genetic model ( OR=0.64 , 95 % CI : 0.44-0.94 ) , the heterozygote genetic model ( OR=0.81 , 95 % CI : 0.69-0.95 ) , and the allelic genetic model ( OR=0.83 , 95 % CI : 0.69-0.99 ) . For R219K in IS , there were significant associations with these genetic models , but not with the recessive genetic model . Our meta-analysis indicated that the O95477 gene R219K polymorphism might be associated with IS and the K allele might be a protective factor in the Chinese population . 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 . 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 .

MH_train_1150

MH_train_1150

MH_train_1150

interacts_with DB00945?

multiple_choice

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 role of cyclooxygenase in gastric mucosal protection . P23219 and P35354 are two cyclooxygenase enzymes responsible for prostanoid production . P35354 is expressed in inflammatory cells and fibroblasts of the gastric mucosa , and through the production of various growth factors including hepatocyte growth factor ( P14210 ) and vascular endothelial growth factor ( P15692 ) , plays a key role in the tissue repair process . DB00945 induces and acetylates P35354 to produce 15-(R)-epi-lipoxinA4 , an anti-inflammatory mediator thought to protect the gastric mucosa against aspirin-induced injury . Recently , three different PGE synthases have been identified , that convert P35354 metabolites into DB00917 . mPGE synthase ( mPGES ) -1 has been shown to be inducible , and to colocalize with P35354 in fibroblasts and macrophages infiltrating the gastric ulcer bed . Q15185 and Q9H7Z7 have been found expressed in normal gastric mucosa , with no change in expression levels seen in gastritis or gastric ulcer tissue . Finally , this review discusses the role of these enzymes in the pathophysiology of the gastric mucosa , as well as the biologcal significance of their inhibition . Vane 's discovery of the mechanism of action of aspirin changed our understanding of its clinical pharmacology . DB00945 exerts its analgesic , antipyretic and anti-inflammatory actions by inhibiting the enzyme cyclooxygenase and thus preventing the formation and release of prostaglandins . The elucidation by John Vane of the mechanism of action of aspirin in 1971 was followed twenty years later by the discovery of a second cyclooxygenase enzyme , P35354 and the rapid development of selective inhibitors of this enzyme . The P35354 inhibitors are potent anti-inflammatory drugs without the damaging side effects on the stomach mucosa of the non-selective aspirin-like inhibitors . More recently , two enzymes have been identified inhibition of which may explain the mechanism of action of paracetamol . These are a putative cyclooxygenase-3 which is a variant of cyclooxygenase-1 and derives from the same gene , and a P35354 variant , induced with diclofenac , which may be involved in the resolution of inflammation . The Effect of Xuefuzhuyu Oral Liquid on DB00945 Resistance and Its Association with rs5911 , rs5787 , and rs3842788 Gene Polymorphisms . DB00945 should be continued indefinitely in patients after interventional therapy , but 10 % to 40 % of patients experience recurrent vascular events despite adequate aspirin therapy , a condition known as aspirin resistance ( AR ) . Xuefuzhuyu oral liquid , derived from the classic recipe Xuefuzhuyu decoction , has been well documented to inhibit platelet aggregation and to improve hemorheology . The aims of this study were to investigate the effects of Xuefuzhuyu oral liquid on AR in patients with chronic stable angina after percutaneous coronary intervention ( P05154 ) and the possible genetic markers related to the drug response . 43 patients diagnosed as having aspirin resistance or semi-resistance were randomly divided into control and treatment groups after screening 207 stable Q8NE62 patients . Platelet aggregation rate was determined using turbidimetry . Three single nucleotide polymorphisms in P23219 ( rs5787 , rs3842788 ) and GP IIb ( rs5911 ) were genotyped in whole blood samples using ABI Q96FA3 7900 HT Fast Real-Time instrument and ABI Q96FA3 3730 DNA Sequencer . The results showed that Xuefuzhuyu oral liquid could effectively improve blood stasis syndrome and AR by inhibiting ADP-induced platelet aggregation and that patients with the rs5911 genetic variant exhibited better drug response upon treatment with Xuefuzhuyu oral liquid , which suggests Xuefuzhuyu oral liquid as a new possible drug for the prevention of AR . 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 . P05305 downregulates angiotensin-converting enzyme-2 expression in human bronchial epithelial cells . BACKGROUND/AIMS : Both endothelin-1 ( ET-1 ) and the renin-angiotensin system ( DB01367 ) are implicated in the pathogenesis and progression of chronic obstructive pulmonary disease ( P48444 ) . In the present study , we explored the interaction between ET-1 and the DB01367 by examining the effect of ET-1 on angiotensin-converting enzyme-2 ( Q9BYF1 ) expression and activity in human bronchial epithelial cells ( HBEpCs ) . METHODS : HBEpCs were treated with ET-1 ( 1 , 10 , 20 , 40 or 50 nmol/l ) for 6 , 12 , 18 , 24 or 30 h with or without the transcription inhibitor actinomycin D , endothelin A ( P25101 ) receptor blocker BQ123 , endothelin B receptor blocker BQ788 , or different kinase inhibitors . RESULTS : ET-1 decreased the Q9BYF1 mRNA level in a dose- and time-dependent manner within 24 h , which led to dose-dependent downregulation of the Q9BYF1 promoter activity , protein level and the cell membrane Q9BYF1 activity . DB00970 ( 1 mg/ml ) , BQ123 ( 1 μmol/l ) , and the p38 mitogen-activated protein kinase ( MAPK ) siRNA and inhibitor PD169316 ( 25 μmol/l ) completely abolished the effect of ET-1 on Q9BYF1 expression in HBEpCs . CONCLUSION : ET-1 downregulates Q9BYF1 expression and activity at the transcription level in HBEpCs via the P25101 receptor by a p38 MAPK-dependent mechanism . This is the first evidence of crosstalk between the ET-1/ P25101 axis and the DB01367 in regard to the pathogenesis and progression of P48444 . 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 . Pharmacokinetic interaction between tadalafil and DB00559 in healthy male subjects . DB00820 , an oral phosphodiesterase 5 ( O76074 ) inhibitor , is being investigated as a treatment for pulmonary arterial hypertension . DB00559 is an oral endothelin receptor antagonist widely used in the treatment of pulmonary arterial hypertension . DB00820 is mainly metabolized by cytochrome P450 ( CYP ) 3A4 , and as DB00559 induces P11712 and P08684 , a pharmacokinetic interaction is possible between these agents . This open-label , randomized study investigated whether any pharmacokinetic interaction exists between tadalafil and DB00559 . Healthy adult men ( n = 15 ; 19-52 years of age ) received 10 consecutive days of tadalafil 40 mg once daily , DB00559 125 mg twice daily , and a combination of both in a 3-period , crossover design . Following 10 days of multiple-dose coadministration of DB00559 and tadalafil , compared with tadalafil alone , tadalafil geometric mean ratios ( 90 % confidence interval [ CI ] ) for AUCtau and Cmax were 0.59 ( 0.55 , 0.62 ) and 0.73 ( 0.68 , 0.79 ) , respectively , with no observed change in tmax . Following coadministration of DB00559 with tadalafil , DB00559 ratios ( 90 % CI ) for AUCtau and Cmax were 1.13 ( 1.02 , 1.24 ) and 1.20 ( 1.05 , 1.36 ) , respectively . DB00820 alone and combined with DB00559 was generally well tolerated . In conclusion , after 10 days of coadministration , DB00559 decreased tadalafil exposure by 41.5 % with minimal and clinically irrelevant differences ( < 20 % ) in DB00559 exposure . 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 . Selective inhibition of prostaglandin endoperoxide synthase-1 ( cyclooxygenase-1 ) by valerylsalicylic acid . DB00945 causes a time-dependent inhibition of prostaglandin endoperoxide H synthases ( PGHS ) -1 and -2 by acetylating active site serines present in both isozymes . In the case of P23219 , aspirin acetylation blocks cyclooxygenase activity , apparently by preventing arachidonate binding to the cyclooxygenase active site . With P35354 , acetylation does not block substrate binding but rather alters the enzyme in such a way that the acetylated form of P35354 produces 15R-hydroxyeicosatetraenoic acid ( 15R-HETE ) instead of the usual prostaglandin endoperoxide product . Based on these differences between P23219 and P35354 , we reasoned that a salicylate ester containing an acyl group somewhat larger than the acetyl group of aspirin might be a selective inhibitor of P35354 . Accordingly , we prepared and tested eight different acyl salicylates as inhibitors of human ( h ) P23219 and -2 expressed transiently in cos-1 cells . Valeryl(pentanoyl)salicylate ( VSA ) was the only compound in this series which showed isozyme selectivity , and , surprisingly , VSA inhibited hPGHS-1 much more effectively than hPGHS-2 . Inhibition of hPGHS-1 by VSA was time-dependent . VSA also inhibited ovine P23219 but did not inhibit the S530A mutant of ovine P23219 . This latter mutant , which lacks the active site serine hydroxyl group , is also refractory to inhibition by acetylsalicylate . Thus , we conclude that VSA acylates the active site serine of P23219 . VSA inhibited prostanoid synthesis by serum-starved murine NIH 3T3 cells which express only P23219 ; in contrast , VSA caused only partial inhibition of prostanoid synthesis by serum-stimulated 3T3 cells which express both PGHS isozymes . Our results establish that VSA can be used as a reasonably selective inhibitor of P23219 . Functional and biochemical evaluation of platelet aspirin resistance after coronary artery bypass surgery . BACKGROUND : DB00945 inhibits platelet activation and reduces atherothrombotic complications in patients at risk of myocardial infarction and stroke . However , a sufficient inhibition of platelet function by aspirin is not always achieved . The causes of this aspirin resistance are unknown . METHODS AND RESULTS : Patients undergoing coronary artery bypass grafting ( CABG ) have a high incidence of aspirin resistance . To evaluate functional and biochemical responses to aspirin , platelet-rich plasma was obtained before and at days 1 , 5 , and 10 after CABG . Thromboxane formation , aggregation , and alpha-granule secretion were effectively inhibited by 30 or 100 micromol/L aspirin in vitro before CABG , but this inhibition was prevented or attenuated after CABG . Whereas the inhibition of thromboxane formation and aggregation by aspirin in vitro partly recovered at day 10 after CABG , oral aspirin ( 100 mg/d ) remained ineffective . The inducible isoform of cyclooxygenase in platelets , P35354 , has been suggested to confer aspirin resistance . In fact , immunoreactive P35354 was increased 16-fold in platelets at day 5 after CABG , but the P35354 selective inhibitor celecoxib did not alter aspirin-resistant thromboxane formation . By contrast , the combined inhibitor of thromboxane synthase and thromboxane receptor antagonist terbogrel equally prevented thromboxane formation of platelets obtained before ( control ) and after CABG . CONCLUSIONS : Platelet aspirin resistance involves an impairment of both in vivo and in vitro inhibition of platelet functions and is probably due to a disturbed inhibition of platelet P23219 by aspirin . DB00945 analogues as dual cyclooxygenase-2/ P09917 inhibitors : synthesis , nitric oxide release , molecular modeling , and biological evaluation as anti-inflammatory agents . Analogues of aspirin were synthesized through an efficient one-step reaction in which the carboxyl group was replaced by an ethyl ester , and/or the acetoxy group was replaced by an N-substituted sulfonamide ( SO(2)NHOR(2):R(2) =H , Me , CH(2)Ph ) pharmacophore . These analogues were designed for evaluation as dual cyclooxygenase-2 ( P35354 ) and P09917 ( 5- P28300 ) inhibitors . In vitro P23219 / P35354 isozyme inhibition studies identified compounds 11 ( CO(2) H , SO(2)NHOH ) , 12 ( CO(2)H , SO(2)NHOCH(2)Ph ) , and 16 ( CO(2)Et , SO(2)NHOH ) as highly potent and selective P35354 inhibitors ( IC(50) range : 0.07-0.7 μM ) , which exhibited appreciable in vivo anti-inflammatory activity ( ED(50) range : 23.1-31.4 mg kg(-1) ) . Moreover , compounds 11 ( IC(50) =0.2 μM ) and 16 ( IC(50) =0.3 μM ) , with a sulfohydroxamic acid ( SO(2)NHOH ) moiety showed potent 5- P28300 inhibitory activity . Furthermore , the SO(2)NHOH moiety present in compounds 11 and 16 was found to be a good nitric oxide ( NO ) donor upon incubation in phosphate buffer at pH 7.4 . Molecular docking studies in the active binding site of P35354 and 5- P28300 provided complementary theoretical support for the experimental biological structure-activity data acquired . P05112 in the Generation of the AERD Phenotype : Implications for Molecular Mechanisms Driving Therapeutic Benefit of DB00945 Desensitization . DB00945 -exacerbated respiratory disease ( AERD ) is explained in part by over-expression of P09917 , leukotriene C4 synthase ( LTC(4)S ) and the cysteinyl leukotriene ( CysLT ) receptors ( CysLT1 and 2 ) , resulting in constitutive over-production of CysLTs and the hyperresponsiveness to CysLTs that occurs with aspirin ingestion . Increased levels of P05112 have been found in the sinus mucosa and nasal polyps of AERD subjects . Previous studies demonstrated that P05112 is primarily responsible for the upregulation of Q16873 by mast cells and the upregulation of CysLT1 and 2 receptors on many immune cell types . Prostaglandin E(2) ( PGE(2) ) acts to prevent CysLT secretion by inhibiting mast cell and eosinophil activation . PGE(2) concentrations are reduced in AERD reflecting diminished expression of cyclooxygenase ( P36551 ) -2 . P05112 can inhibit basal and stimulated expression of P35354 and microsomal PGE synthase 1 leading to decreased capacity for PGE(2) secretion . Thus , P05112 plays an important pathogenic role in generating the phenotype of AERD . This review will examine the evidence supporting this hypothesis and describe a model of how aspirin desensitization provides therapeutic benefit for AERD patients . DB00945 -responsive , migraine-like transient cerebral and ocular ischemic attacks and erythromelalgia in O60674 -positive essential thrombocythemia and polycythemia vera . Migraine-like cerebral transient ischemic attacks ( MIAs ) and ocular ischemic manifestations were the main presenting features in 10 O60674 (V617F)-positive patients studied , with essential thrombocythemia ( ET ) in 6 and polycythemia vera ( PV ) in 4 . Symptoms varied and included cerebral ischemic attacks , mental concentration disturbances followed by throbbing headaches , nausea , vomiting , syncope or even seizures . MIAs were frequently preceded or followed by ocular ischemic events of blurred vision , scotomas , transient flashing of the eyes , and sudden transient partial blindness preceded or followed erythromelalgia in the toes or fingers . The time lapse between the first symptoms of aspirin-responsive MIAs and the diagnosis of ET in 5 patients ranged from 4 to 12 years . At the time of erythromelalgia and MIAs , shortened platelet survival , an increase in the levels of the platelet activation markers β-thromboglobulin and platelet factor 4 and also in urinary thromboxane B2 were clearly indicative of the spontaneous in vivo platelet activation of constitutively O60674 (V617F)-activated thrombocythemic platelets . DB00945 relieves the peripheral , cerebral and ocular ischemic disturbances by irreversible inhibition of platelet cyclo-oxygenase ( P23219 ) activity and aggregation ex vivo . Vitamin K antagonist , dipyridamole , ticlopidine , sulfinpyrazone and sodium salicylate have no effect on platelet P23219 activity and are ineffective in the treatment of thrombocythemia-specific manifestations of erythromelalgia and atypical MIAs . If not treated with aspirin , ET and PV patients are at a high risk of major arterial thrombosis including stroke , myocardial infarction and digital gangrene . Effect of cyclooxygenase-2 inhibitor ( celecoxib ) on the infarcted heart in situ . Several attempts have been made to replace aspirin with compounds without gastric toxicity ; a cyclooxygenase-2 ( P35354 ) inhibitor , celecoxib , and a nitric oxide-aspirin , O43763 -4016 , have been developed for this purpose . This paper compares effects of celecoxib , O43763 -4016 and aspirin on production of prostacyclin ( DB01240 ) and thromboxane A2 ( TXA2 ) and activation of the inducible form of nitric oxide synthase ( P35228 ) in infarcted heart in situ . DB00945 was most effective in reducing myocardial DB01240 synthesis and formation of TXA2 . Myocardial effects of celecoxib resemble those of O43763 -4016 , although the two compounds have different modes of action . 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 . DB00945 insensitive eicosanoid biosynthesis in cardiovascular disease . Inhibition of platelet cyclooxygenase ( P36551 ) -1 is involved in aspirin cardioprotection observed in clinical trials , but , in some patients , aspirin is unable to protect from thrombotic complications . An incomplete suppression of platelet thromboxane ( TX ) A2 biosynthesis has been assumed to participate in the phenomenon of aspirin resistance , as a consequence of the following possible mechanisms : ( i ) P35354 expression in newly formed platelets ; ( ii ) pharmacodynamic interactions between aspirin and coadministered nonsteroidal antiinflammatory drugs ( e.g. ibuprofen ) ; ( iii ) expression of variant isoforms of P23219 with reduced sensitivity to irreversible inactivation at Ser529 . Furthermore , aspirin failure may be due to enhanced formation of vasoactive and/or proaggregatory eicosanoids despite an almost complete suppression of platelet TXA2 biosynthesis by aspirin . Thus , in a subset of patients with unstable angina treated with low-dose aspirin , to almost completely block platelet P23219 activity , enhanced TXA2 biosynthesis in vivo has been demonstrated , presumably through an increased generation of P35354 -dependent PGH2 in plaque monocytes/macrophages or activated vascular cells . The concomitant increased formation of 8-iso-PGF2alpha , one of the most abundant F2-isoprostanes in humans , generated by free-radical catalyzed arachidonate peroxidation , may be involved in aspirin resistance because of its capacity to induce platelet and vascular smooth muscle cell activation through the interaction with thromboxane receptors ( TPs ) . Finally , enhanced production of vasoactive cysteinyl leukotrienes ( cys-LTs ) occurs in unstable angina despite conventional antithrombotic and antianginal treatment . The use of selective pharmacological tools ( i.e. highly selective P35354 inhibitors , TP antagonists , cys-LT inhibitors and antagonists ) will allow to ascertain the contribution of aspirin insensitive eicosanoid biosynthesis in aspirin cardioprotective failure . Roles of platelet and endothelial cell P23219 in hypercholesterolemia-induced microvascular dysfunction . DB00945 is a common preventative therapy in patients at risk for cardiovascular diseases , yet little is known about how aspirin protects the vasculature in hypercholesterolemia . The present study determines whether aspirin , nitric oxide-releasing aspirin ( O43763 -4016 ) , a selective cyclooxygenase ( P36551 ) -1 inhibitor ( SC560 ) , or genetic deficiency of P23219 prevents the inflammatory and prothrombogenic phenotype assumed by hypercholesterolemic ( HC ) venules . DB00945 or O43763 -4016 ( 60 mg/kg ) was administered orally for the last week of a 2-wk HC diet . P23219 -deficient ( P23219 (-/-) ) and wild-type ( WT ) mice were transplanted with WT ( WT/ P23219 (-/-) ) or P23219 (-/-) ( P23219 (-/-)/WT ) bone marrow , respectively . HC-induced adhesion of platelets and leukocytes in murine intestinal venules , observed with intravital fluorescence microscopy , was greatly attenuated in aspirin-treated mice . Adhesion of aspirin-treated platelets in HC venules was comparable to untreated platelets , whereas adhesion of SC560-treated platelets was significantly attenuated . HC-induced leukocyte and platelet adhesion in P23219 (-/-)/WT chimeras was comparable to that in SC560-treated mice , whereas the largest reductions in blood cell adhesion were in WT/ P23219 (-/-) chimeras . O43763 -4016 treatment of platelet recipients or donors attenuated leukocyte and platelet adhesion independent of platelet P23219 inhibition . Platelet- and endothelial cell-associated P23219 promote microvascular inflammation and thrombogenesis during hypercholesterolemia , yet nitric oxide-releasing aspirin directly inhibits platelets independent of P23219 . 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 . DB00945 induction of apoptosis in esophageal cancer : a potential for chemoprevention . The potential use of non-steroidal anti-inflammatory drugs ( NSAIDs ) in the prevention of gastrointestinal cancers has been highlighted recently . However , it is not known whether NSAIDs could also be useful for preventing esophageal cancer , although regular users of these drugs appear to have a decreased incidence of esophageal cancer . Therefore , we examined the effect of aspirin on growth and apoptosis in 10 esophageal cancer cell lines as well as the expression and modulation of its target enzymes , cyclooxygenases ( COXs ) , and their product prostaglandin E2 . Growth inhibition of these cells by aspirin was dose- and time-dependent and associated with the induction of apoptosis . P23219 and P35354 were expressed in 7 of the 10 cell lines . Bile acids could induce P35354 expression in six of eight cell lines tested , which was correlated with prostaglandin E2 production , and aspirin could inhibit P35354 enzymatic activity even after bile acid stimulation but was unable to change the P35354 protein level in these cell lines . Down-regulation of bcl-2 by aspirin was found in the two cell lines tested . These results suggest that induction of apoptosis by aspirin may be a mechanism by which it can intervene in esophageal carcinogenesis and may be indicative of the potential of NSAIDs as chemopreventive agents in esophageal cancer . DB00945 , NSAIDs , and P35354 inhibitors in cardiovascular disease : possible interactions and implications for treatment of rheumatoid arthritis . DB00945 , nonsteroidal antiinflammatory drugs ( NSAIDs ) , and cyclooxygenase-2 ( P35354 ) inhibitors are widely used in patients with rheumatoid arthritis . DB00945 has the largest and most persuasive body of randomized trial evidence to support its use in secondary prevention for cardiovascular disease ( CVD ) and primary prevention for myocardial infarction . There is , however , a possible deleterious interaction between aspirin and NSAIDs on CVD that requires further research . DB00945 , NSAIDs , and to a lesser extent P35354 inhibitors are associated with increased gastrointestinal side effects and bleeding , alone and in combination . The more widespread and appropriate use of aspirin in patients with rheumatoid arthritis will avoid many premature deaths in secondary prevention for CVD and first myocardial infarctions in primary prevention . 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 . P35354 induction and prostaglandin E2 accumulation in squamous cell carcinoma as a consequence of epidermal growth factor receptor activation by imatinib mesylate . Imatinib mesylate is a novel anti-tumor agent useful in the clinical management of chronic myelogenous leukemia and gastrointestinal stromal tumors with minimal toxicity relative to other forms of cancer therapy . Its clinical activity and minimal toxicity are related to specific inhibition of cellular targets including P11274 - P00519 , platelet-derived growth factor receptor and c-kit kinases , resulting in the collapse of downstream signaling cascades important for transformation . In some patients , unexpected toxicities arise that are not associated with inhibition of any known cellular imatinib target . In this report , we investigated the effects of imatinib on squamous carcinoma cell signaling . Imatinib induced expression of P35354 in a dose-dependent manner with concomitant accumulation of prostaglandin E2 . P35354 induction by imatinib was initiated through epidermal growth factor ( P01133 ) receptor kinase activation and downstream signaling through mitogenic-activated protein kinase . P35354 induction by imatinib was blocked by Q02750 or P01133 receptor inhibition . Imatinib did not activate stressor cytokine-signaling pathways ( p38 kinase , nuclear factor-kB nuclear translocation ) or affect P23219 expression . Imatinib failed to activate P01133 receptor signals in other tumor types , suggesting that P35354 induction in imatinib-treated cells is mediated through release of autocrine factors expressed or activated in squamous tumors . P35354 induction by imatinib in squamous tumors derived from the head and neck region is unique with respect to other target-specific agents and may represent one of the unintended toxic effects of imatinib described in some patients . DB00945 and sodium salicylate inhibit endothelin P25101 receptors by an allosteric type of mechanism . DB00945 is a commonly used drug with a wide pharmacological spectrum including antiplatelet , anti-inflammatory , and neuroprotective actions . This study shows that aspirin and sodium salicylate , its major blood metabolite , reverse contractile actions of endothelin-1 ( ET-1 ) in isolated rat aorta and human mammary arteries . They also prevent the intracellular Ca(2+) mobilizing action of ET-1 in cultured endothelial cells but not those of neuromedin B or UTP . Inhibition of the actions of ET-1 by salicylates is apparently competitive . Salicylates inhibit (125)I-ET-1 binding to recombinant rat P25101 receptors . Salicylic acid promotes dissociation of (125)I-ET-1 P25101 receptor complexes both in the absence and the presence of unlabeled ET-1 . It has no influence on the rate of association of (125)I-ET-1 to P25101 receptors . Salicylates do not promote dissociation of (125)I-ET-1 ETB receptor complexes . Salicylates potentiate relaxing actions of receptor antagonists such as DB00559 . It is concluded that salicylates are allosteric inhibitors of P25101 receptors . The results also suggest that : 1 ) irreversible ET-1 binding probably limits actions of receptor antagonists in vivo , and 2 ) an association of salicylates and P25101 receptor antagonists should be used to evaluate the physiopathological role of ET-1 and may be of therapeutic interest in the treatment of ischemic heart disease . Non-steroidal anti-inflammatory drug and aspirin use and the risk of head and neck cancer : a systematic review . BACKGROUND : Use of non-steroidal anti-inflammatory drugs ( NSAIDs ) has been associated with a reduced risk of several cancers . This is thought to be through the inhibitory action on the cyclooxygenase ( P36551 ) enzyme , P35354 . Evidence for NSAIDs preventing head and neck cancer ( HNC ) is conflicting . We conducted a systematic literature review to investigate the association between NSAID/aspirin use and risk of head and neck cancer ( HNC ) . METHODOLOGY : MEDLINE , EMBASE , PubMed , Cochrane Library , and Web of Science were systematically searched using terms for NSAIDs/aspirin , HNC , and observational/intervention study designs to identify studies published by December 2009 . RESULTS : Of 9,268 articles identified , two population-based prescribing database studies and three case-control studies met the selection criteria . The studies investigated different HNC sites . Only one study found a significant protective association of aspirin use with HNC risk ( OR 0.75 , 95 % CI 0.58-0.96 ) , and one showed a significantly increased risk of oral/oropharyngeal cancer with non-low-dose aspirin NSAID use ( OR 3.5 , 95 % CI 1.8-6.7 ) . Many of the studies identified lacked information on important confounding factors . CONCLUSION : No definitive conclusion on the effect of NSAIDs/aspirin on HNC risk was possible . DB00945 may protect against HNC , although further robust large-scale studies are required to clarify any possible association . 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 . 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 . Tyrosine phosphorylation of Q92835 promotes its proteasomal degradation . OBJECTIVE : The activity of the SH2-containing-phosphatidylinositol-5'-phosphatase ( Q92835 , also known as Q92835 ) , a critical hematopoietic-restricted negative regulator of the P19957 kinase ( PI3K ) pathway , is regulated in large part via its protein levels . We sought to determine the mechanism(s) involved in its downregulation by P11274 - P00519 and by interleukin ( IL ) -4 . MATERIALS AND METHODS : We used Ba/ P13726 ( Q92817 -tetOFF) cells to study the downregulation of Q92835 by P11274 - P00519 and bone marrow-derived macrophages to study Q92835 's downregulation by P05112 . RESULTS : We show herein that P11274 - P00519 downregulates Q92835 , but not O15357 or P60484 , and this can be blocked with the Src kinase inhibitor Q99463 , which inhibits the tyrosine phosphorylation of Q92835 , or with the proteasomal inhibitor MG-132 . We also show , using anti- Q92835 immunoprecipitates , that c-Cbl and Cbl-b are associated with Q92835 and that P11274 - P00519 induces Q92835 's polyubiquitination . This ubiquitination can be blocked with Q99463 , consistent with the tyrosine phosphorylation of Q92835 acting as a signal for its ubiquitination . In bone marrow-derived macrophages , P05112 also leads to the proteasomal degradation of Q92835 but , unlike in Ba/ P13726 ( Q92817 -tetOFF) cells , O15357 is also proteasomally degraded and the degradation of both inositol phosphatases can be prevented with Q99463 or MG-132 . CONCLUSION : Our results suggest that Q92835 protein levels can be reduced via P11274 - P00519 and/or Src family member-induced tyrosine phosphorylation of Q92835 because this triggers its polyubiquitination and degradation within the proteasome . 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 . 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 . [ Prevention of thrombosis and vascular inflammation : importance of combined cyclooxygenase and P09917 inhibitors ] . DB00945 , a standard non-steroidal anti-inflammatory drug ( NSAID ) is currently used in antithrombotic treatment . However , its use is limited by largely recognized gastrotoxicity and recommended doses are low . The major side effect of aspirin is related to its ability to suppress prostaglandin ( PG ) synthesis by constitutive cyclooxygenase-1 ( P23219 ) . Specific inhibitors of P35354 , the inducible isoform of P36551 which was more recently described , have potent antiinflammatory effects . They are associated with minor risk of gastric tractus toxicity and reduced inflammatory leukocyte components known for their proatherothrombotic properties . Nevertheless , recent findings attributed a significant cardiovascular risk to some of them . P09917 ( 5- P28300 ) , an enzyme mainly expressed by leukocytes , is responsible for the generation of leukotrienes , the major lipidic proinflammatory mediators . Development of combined inhibitors of 5- P28300 and P36551 isoforms 1 and 2 inaugurate an interesting new therapeutic pathway . Indeed , such inhibitors suppress not only the activation of platelets , leukocytes and endothelial cells but also prevent their metabolic and functional interactions . In addition to their broad spectrum inhibition , they may be associated with the minor gastrotoxic effect . Thus , platelet-leukocyte interactions which dominate the underlying inflammatory process particularly in atherosclerosis , might reinforce the benefits of such inhibitors . DB00945 and age-related macular degeneration : positives versus negatives . The anti-inflammatory , analgesic , antipyretic and antithrombotic activities of aspirin confer its wide therapeutic application . The three former activities require higher doses of aspirin , whereas the latter can be achieved through a lower , thus safer dose of the drug . Low-dose , long-term aspirin is used as an antithrombotic therapy to prevent cardiovascular disease . Such therapy is used by millions of people worldwide , including those suffering from age-related macular degeneration ( AMD ) ; thus , questions have arisen as to whether such treatment has any impact on the development and course of AMD . This editorial addresses the important issue of possible beneficial and adverse effects of long-term , low-dose aspirin treatment of AMD patients . Special emphasis is given to the ability of aspirin to acetylate cyclooxygenases ( especially P35354 ) and thus to initiate a biochemical pathway leading to the generation of anti-inflammatory pro-resolving mediators synthesized from both ω-3 and ω-6 long-chain polyunsaturated fatty acids . Such mediators ( e.g. , resolvins , lipoxins ) may be of therapeutic value in retarding the development of dry form AMD . Nonsteroidal antiinflammatory drugs and cyclooxygenase inhibition in the gastrointestinal tract : a trip from peptic ulcer to colon cancer . DB00945 was commercialized more than a 100 years ago . Today , this compound is still widely prescribed , and new mechanisms of action and indications are being tested . Inhibition of cyclooxygenase ( P36551 ) -1 and P35354 by aspirin or its related compounds , nonsteroidal antiinflammatory drugs ( NSAIDs ) , has been associated with both adverse and beneficial effects in the gastrointestinal ( GI ) tract . Inhibition of P23219 has been linked to GI adverse effects . Adverse effects of NSAIDs and aspirin in the upper GI tract include esophagitis , peptic ulcer , peptic ulcer complications , and death . Effective preventive therapies are available that have been associated with a progressive decline in the rate of hospitalization due to upper GI complications . NSAIDs and aspirin can also damage the small bowel and the colon . NSAID enteropathy is frequent and in most cases subclinical ( increased mucosal permeability , inflammation , erosion , ulcer ) . However , more serious clinical outcomes such as anemia , bleeding , perforation , obstruction , diverticulitis , and deaths have also been described . Prevention therapy of NSAID damage to the lower GI tract is not well defined . Inhibition of P35354 by NSAIDs , coxibs , or aspirin seems to provide beneficial effects to the GI tract . Observational studies show that these compounds reduce the risk of both upper and lower GI cancers . Randomized controlled trials have shown that aspirin and coxibs reduce the recurrence rate of colonic polyps , and long-term cohort studies have shown that aspirin reduces the risk of colon cancer time and dose dependently . New studies will have to define the appropriate population that may benefit with these therapies . The comparative study of acetyl-11-keto-beta-boswellic acid ( AKBA ) and aspirin in the prevention of intestinal adenomatous polyposis in P25054 (Min/+) mice . Acetyl-11-keto-beta-BA ( AKBA ) , a component of the gum resin of Boswellia serrata , has been recognized as a promising agent for the prevention of intestinal tumorigenesis . DB00945 , a non-steroidal anti-inflammatory drug ( NSAID ) , has also been considered to have the activity against intestinal tumorigenesis . However , the prevention of colonic cancer is insufficient and no definitive recommendation has been made for clinic use . Herein , we compared the efficacy of AKBA with that of aspirin in an adenomatous polyposis coli intestinal neoplasia consecutive weeks . Mice were sacrificed by anesthetizing . The whole intestine was removed from each mouse . The number , size and histopathology of intestinal adenomatous polyps were examined under microscopy . The adenomatous polyps were removed for further analysis by the assays of western blotting and immunohistochemical staining . AKBA significantly prevented the formation of intestinal adenomatous polyps without toxicity to mice . Statistical analysis indicated that AKBA 's activity both in the prevention of small intestinal and colonic polyps was more potently than aspirin . Histopathologic examination revealed that AKBA 's effect , that is the reduction of polyp size and degree of dysplasia , was more prominent in larger sized polyps , especially those originating in colon . These effects of AKBA were associated with its role in the induction of apoptosis in carcinomas . The assays of western blotting and immunohistochemistry staining indicated that the efficacy of AKBA might arise from its activity in the modulation of the Wnt/β-catenin pathway and NF-κB/ P35354 pathway in adenomatous polyps . Conclusion , AKBA by oral application prevented intestinal tumorigenesis more potential than aspirin . 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 . 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 . Interaction between P20292 and P20815 gene variants significantly increases the risk for cerebral infarctions in Chinese . In this study , we investigated associations between susceptibility genes and cerebral infarctions in a Chinese population , and whether gene-gene interactions increase the risk of cerebral infarctions . Overall , 292 patients with cerebral infarctions and 259 healthy control individuals were included . Eight variants in five candidate genes were examined for the risk of stroke , including the SG13S32 ( rs9551963 ) , SG13S42 ( rs4769060 ) , SG13S89 ( rs4769874 ) , and SG13S114 ( rs10507391 ) variants of the P09917 activating protein ( P20292 ) gene , the G860A ( rs751141 ) variant of the soluble epoxide hydrolase ( P34913 ) gene , the A1075C ( rs1057910 ) variant of the P11712 *2 gene , the C430T ( rs1799853 ) variant of the P11712 *3 gene , and the A6986G ( rs776746 ) variant of the P20815 gene . Gene-gene interactions were explored using generalized multifactor dimensionality reduction methods . There were no statistically significant differences in the frequencies of the genotypes of the eight candidate genes . The generalized multifactor dimensionality reduction analysis showed a significant gene-gene interaction between SG13S114 and A6986G , with scores of 10 for cross-validation consistency and 9 for the sign test ( P=0.0107 ) . These gene-gene interactions predicted a significantly higher risk of cerebral infarction ( adjusted for age , hypertension , and diabetes mellitus ; odds ratio=1.80495 % , confidence interval : 1.180-2.759 , P=0.006 ) . A two-loci gene interaction confers a significantly higher risk for cerebral infarction . The combinational analysis used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases . 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 . 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 . Stereospecificity of hydrogen abstraction in the conversion of arachidonic acid to 15R-HETE by aspirin-treated cyclooxygenase-2 . Implications for the alignment of substrate in the active site . The initial and rate-limiting step in prostaglandin biosynthesis is stereoselective removal of the pro-S hydrogen from the 13-carbon of arachidonic acid . This is followed by oxygenation at C-11 , formation of the five-membered ring , and a second oxygenation at C-15 to yield the endoperoxide product , prostaglandin G(2) . DB00945 treatment of cyclooxygenase-2 is known to acetylate an active site serine , block prostaglandin biosynthesis , and give 15R-hydroxyeicosatetraenoic acid ( 15R-HETE ) as the only product . 15R-HETE and prostaglandins have opposite stereoconfigurations of the 15-hydroxyl . To understand the changes that lead to 15R-HETE synthesis in aspirin-treated P35354 , we employed pro-R- and pro-S-labeled [13-(3)H]arachidonic acids to investigate the selectivity of the initial hydrogen abstraction . Remarkably , aspirin-treated P35354 formed 15R-HETE with removal of the pro-S hydrogen at C-13 ( 3-9 % retention of pro-S tritium label ) , the same stereoselectivity as in the formation of prostaglandins by native cyclooxygenase . To account for this result and the change in oxygenase specificity , we suggest that the bulky serine acetyl group forces a realignment of the omega end of the arachidonic acid carbon chain . This can rationalize abstraction of the C-13 pro-S hydrogen , the blocking of prostaglandin synthesis , and the formation of 15R-HETE as the sole enzymatic product . [ 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 . 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 . 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 . The anti-tumor effect of HDAC inhibition in a human pancreas cancer model is significantly improved by the simultaneous inhibition of cyclooxygenase 2 . Pancreatic ductal adenocarcinoma is the fourth leading cause of cancer death worldwide , with no satisfactory treatment to date . In this study , we tested whether the combined inhibition of cyclooxygenase-2 ( P35354 ) and class I histone deacetylase ( HDAC ) may results in a better control of pancreatic ductal adenocarcinoma . The impact of the concomitant HDAC and P35354 inhibition on cell growth , apoptosis and cell cycle was assessed first in vitro on human pancreas BxPC-3 , PANC-1 or CFPAC-1 cells treated with chemical inhibitors ( DB02546 , MS-275 and celecoxib ) or Q13547 /2/3/7 siRNA . To test the potential antitumoral activity of this combination in vivo , we have developed and characterized , a refined chick chorioallantoic membrane tumor model that histologically and proteomically mimics human pancreatic ductal adenocarcinoma . The combination of Q13547 /3 and P35354 inhibition significantly impaired proliferation of BxPC-3 cells in vitro and stalled entirely the BxPC-3 cells tumor growth onto the chorioallantoic membrane in vivo . The combination was more effective than either drug used alone . Consistently , we showed that both Q13547 and O15379 inhibition induced the expression of P35354 via the NF-kB pathway . Our data demonstrate , for the first time in a Pancreatic Ductal Adenocarcinoma ( PDAC ) model , a significant action of HDAC and P35354 inhibitors on cancer cell growth , which sets the basis for the development of potentially effective new combinatory therapies for pancreatic ductal adenocarcinoma patients . Monoclonal antibodies against human dendritic cell-like peripheral blood monocytes activated by granulocyte/macrophage-colony-stimulating factor plus interleukin 4 . Human peripheral blood monocytes activated by GM- P04141 plus P05112 have recently been found to exhibit characteristics of putative dendritic cells ( DC ) . These cytokine-activated monocytes ( P62158 ) may express novel activation Ag that contribute significantly to their antigen presentation potency . To examine that possibility , mAb specific for P62158 were derived . Seven mAb that stained P62158 but not unactivated monocytes and other peripheral blood mononuclear cell types were identified . Further screening with a panel of cell lines identified two P62158 -specific mAb . The first mAb , 2.1D10 , was found to be mannose-receptor specific . A second mAb , 6.3B7 , immunoprecipitated a 190-kDa Ag . It stained neither activated B cells nor the putative peripheral blood precursor DC population . Furthermore , 6.3B7 did not recognize determinants in asparagine-linked carbohydrate chains or in sialic acid-containing structures . These mAb against P62158 membrane proteins may provide new insights into the requirements for optimal antigen presentation by macrophages and other P25054 types . 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 . 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 . 12-hydroxyeicosatetraenoic acid is associated with variability in aspirin-induced platelet inhibition . BACKGROUND : DB00945 is one of the most widely used non-steroidal anti-inflammatory drugs ( NSAIDs ) . It is also a commonly used anti-platelet drug , which inhibits the formation of the platelet activator , thromboxane A2 ( TxA2 ) via inhibition of cyclooxygenase-1 ( P23219 ) . However , the presence of a patient subset that fails to respond to aspirin despite reduced TxA2 concentrations suggests that the effect of aspirin might be more complex than exclusive P23219 inhibition . METHODS : In this study we evaluated the impact of in vivo oral administration of a standard anti-platelet dose ( 75 mg ) of aspirin in healthy volunteers on the acute impact of in vitro collagen-mediated platelet aggregation and generation of platelet-derived TxA2 and the 12-lipoxygenase ( P28300 ) metabolite 12-hydroxyeicosatetraenoic acid ( 12-HETE ) . The eicosanoids were quantified using liquid chromatography-tandem mass spectrometry ( LC-MS/MS ) . RESULTS : Low-dose aspirin administration not only inhibited TxA2 generation but also decreased the production of 12-HETE . Furthermore , a significant correlation was observed between the levels of 12-HETE and collagen-induced platelet aggregation . Pre-treatment of platelets with the P16050 inhibitor , baicalein , prior to activation attenuated platelet aggregation . CONCLUSIONS : These findings support a role for 12-HETE as a pro-aggregatory eicosanoid in platelet function and suggest a role for 12-HETE in variable sensitivity to aspirin . The study also highlights a potentially important mechanism by which aspirin impacts upon eicosanoid generation . DB00945 -induced peptic ulcer and genetic polymorphisms . There are a few studies of the association between genetic polymorphisms and the risks of acetylsalicylic acid (aspirin)-induced ulcer or its complications . Two single nucleotide polymorphisms ( SNP ) of cyclooxygenase-1 ( P23219 ) , A-842G and C50T , exhibited increased sensitivity to aspirin and had lower prostaglandin synthesis capacity , lacking statistical significance in the association with bleeding peptic ulcer . A recent Japanese study indicated that the number of P23219 -1676T alleles was a significant risk factor for peptic ulcer in users of non-steroidal anti-inflammatory drugs ( NSAIDs ) . There are some genetic polymorphisms for aspirin resistance , such as platelet membrane glycoproteins , thromboxane A2 ( TXA2 ) receptor , platelet activating factor acetylhydrolase and coagulation factor XIII ; however , data on the frequency of gastrointestinal ( GI ) events in these variants are lacking . Carrying the P11712 variants is reported a significantly increased risk of non-aspirin NSAID-related GI bleeding . The polymorphisms of interleukin-1beta ( IL-1beta ) and tumor necrosis factor-alpha ( P01375 ) have been associated with development of peptic ulcer or gastric cancer . In a recent investigation , carriage of the IL-1beta-511 T allele was significantly associated with peptic ulcer among low-dose aspirin users . Hypoacidity in corpus gastritis related to polymorphisms of pro-inflammatory cytokines seems to reduce NSAIDs or aspirin-related injury . Data on which polymorphisms are significant risk factors for GI events in aspirin users are still lacking and further large-scale clinical studies are required . Human mesenchymal stem cell proliferation is regulated by DB00917 through differential activation of DB02527 -dependent protein kinase isoforms . The conditions used for in vitro differentiation of hMSCs contain substances that affect the activity and expression of cyclooxygenase enzymes ( P23219 / P35354 ) and thereby the synthesis of prostanoids . hMSC constitutively produce DB00917 when cultivated in vitro . In this study we have investigated effects of DB00917 on proliferation of hMSC . We here demonstrate that one of the main control molecules in the Wnt pathway , P49841 , is phosphorylated at the negative regulatory site ser-9 after treating the cells with DB00917 . This phosphorylation is mediated by elevation of DB02527 and subsequent activation of PKA . Furthermore , DB00917 treatment leads to enhanced nuclear translocation of beta-catenin , thus influencing cell proliferation . The presence of two PKA isoforms , types I and II , prompted us to investigate their individual contribution in DB00917 -mediated regulation of proliferation . Specific activation of PKA type II with synthetic DB02527 analogues , resulted in enhancement of proliferation . On the other side , we found that treatment of hMSC with high concentrations of DB00917 inhibited cell proliferation by arresting the cells in G0/ P55008 phase , an effect we found to be mediated by PKA I . Hence , the two different PKA isoforms seem to have opposing functions in the regulation of proliferation and differentiation in these cells . 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 . DB00945 and stroke prevention . According to meta-analyses aspirin provides a relative reduction in the rate of major vascular events of 19 % in patients with arterial disease in general , whereas for patients with ischaemic cerebrovascular disease this reduction is only 13 % . The discrepancy may well result from pathophysiological differences and not from a play of chance . There is no proven difference in efficacy according to dose . The evidence for this equivalence is most compelling in the range between 75 and 1300 mg daily , but still fairly convincing for doses between 30 and 50 mg . In contrast , side effects are clearly more frequent as the dose is higher . Other antiplatelet agents ( sulfinpyrazone , ticlopidine , clopidogrel , dipyridamole , orally administered IIb/IIIa inhibitors ) have no clear advantages over aspirin and in some cases definite disadvantages ; the combination of aspirin and dipyridamole may be more efficacious than aspirin alone , but the evidence hinges on a single trial . If recurrent TIAs occur under treatment with aspirin , the rational response is not to change to a different antiplatelet agent , but to review the diagnosis and consider causes other than artery-to-artery embolism . Platelet aggregation can probably still occur despite complete acetylation of platelets , via pathways other than P23219 inhibition , but in vitro aggregation tests are an unreliable measure .

MH_train_1151

MH_train_1151

MH_train_1151

interacts_with DB00945?

multiple_choice

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 . Prostanoid receptor antagonists : development strategies and therapeutic applications . Identification of the primary products of cyclo-oxygenase ( P36551 ) /prostaglandin synthase(s) , which occurred between 1958 and 1976 , was followed by a classification system for prostanoid receptors ( DP , EP(1) , EP(2) ... ) based mainly on the pharmacological actions of natural and synthetic agonists and a few antagonists . The design of potent selective antagonists was rapid for certain prostanoid receptors ( EP(1) , TP ) , slow for others ( FP , IP ) and has yet to be achieved in certain cases ( EP(2) ) . While some antagonists are structurally related to the natural agonist , most recent compounds are ' non-prostanoid ' ( often acyl-sulphonamides ) and have emerged from high-throughput screening of compound libraries , made possible by the development of ( functional ) assays involving single recombinant prostanoid receptors . Selective antagonists have been crucial to defining the roles of P52209 (2) ( acting on DP(1) and DP(2) receptors ) and PGE(2) ( on EP(1) and EP(4) receptors ) in various inflammatory conditions ; there are clear opportunities for therapeutic intervention . The vast endeavour on TP ( thromboxane ) antagonists is considered in relation to their limited pharmaceutical success in the cardiovascular area . Correspondingly , the clinical utility of IP ( prostacyclin ) antagonists is assessed in relation to the cloud hanging over the long-term safety of selective P35354 inhibitors . DB00945 apart , P36551 inhibitors broadly suppress all prostanoid pathways , while high selectivity has been a major goal in receptor antagonist development ; more targeted therapy may require an intermediate position with defined antagonist selectivity profiles . This review is intended to provide overviews of each antagonist class ( including prostamide antagonists ) , covering major development strategies and current and potential clinical usage . 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 . 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 . 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 . 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 . Analgesics . DB00945 and other non-steroidal anti-inflammatory drugs ( NSAIDs ) are often incriminated in hypersensitivity reactions leading to anaphylaxis . Two populations are at the high risk of developing such reactions : patients with asthma and those with urticaria . In a subset of asthmatics , NSAIDs that inhibit cyclooxygenase-1 ( P23219 ) precipitate non-allergic , hypersensitivity reactions , characterized by violent attacks of dyspnea . These patients suffer from a distinct clinical syndrome , called aspirin induced asthma ( AIA ) , which includes chronic eosinophilic rhinusinusitis and persistent asthma . Inpatients with chronic idiopathic urticaria , and less commonly in patients without chronic urticaria , NSAIDs usually acting through inhibition of P23219 can induce or exacerbate skin eruptions . While alterations in eicosanoid biosynthesis characterized both AIA and aspirin-triggered urticaria , other patients may rarely manifest IgE-mediated reactions . 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 . 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 . DB00945 and NS-398 inhibit hepatocyte growth factor-induced invasiveness of human hepatoma cells . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) inhibit cyclooxygenase ( P36551 ) activity and are considered to exert antitumor actions in a variety of cancer cells , although the effects are unlikely entirely due to P36551 inhibition . Because clinical observations suggest that hepatocyte growth factor ( P14210 ) can promote metastasis of hepatoma cells while stimulating tumor invasiveness , we investigated the effect of aspirin and NS-398 , a selective P35354 inhibitor , on P14210 -mediated invasiveness of HepG2 human hepatoma cells . P14210 stimulated the invasiveness of HepG2 cells in Matrigel cell invasion assay , together with increased expression of matrix metalloproteinase ( MMP ) 9 . Addition of aspirin or NS-398 , similar to PD98059 , which acts as a specific inhibitor of mitogen-activated protein kinase/extracellular signal-regulated kinase ( MEK ) , an upstream kinase regulating extracellular signal-regulated kinase ( P29323 )1/2 , abrogated such actions of P14210 without affecting cell viability . DB00945 and NS-398 , in contrast to PD98059 , did not suppress P27361 /2 phosphorylation induced by P14210 . However , both agents inhibited the kinase activity of P27361 /2 induced by P14210 and repressed P14210 -induced phosphorylation of 90-kd ribosomal S6 kinase ( RSK ) and Elk-1 , key downstream substrates of P27361 /2 , resulting in the suppression of transcriptional activity of Elk-1 as well as nuclear factor kappaB ( NF-kappaB ) and AP-1 , which are involved in P14780 gene regulation . In conclusion , our results suggest that aspirin and NS-398 inhibit P14210 -induced invasiveness of HepG2 human hepatoma cells through P27361 /2 . DB00945 , cyclooxygenase inhibition and colorectal cancer . Colorectal cancer ( CRC ) is the third most common type of cancer worldwide . Screening measures are far from adequate and not widely available in resource-poor settings . Primary prevention strategies therefore remain necessary to reduce the risk of developing CRC . Increasing evidence from epidemiological studies , randomized clinical trials and basic science supports the effectiveness of aspirin , as well as other non-steroidal anti-inflammatory drugs , for chemoprevention of several types of cancer , including CRC . This includes the prevention of adenoma recurrence and reduction of CRC incidence and mortality . The detectable benefit of daily low-dose aspirin ( at least 75 mg ) , as used to prevent cardiovascular disease events , strongly suggests that its antiplatelet action is central to explaining its antitumor efficacy . Daily low-dose aspirin achieves complete and persistent inhibition of cyclooxygenase ( P36551 ) -1 in platelets ( in pre-systemic circulation ) while causing a limited and rapidly reversible inhibitory effect on P35354 and/or P23219 expressed in nucleated cells . DB00945 has a short half-life in human circulation ( about 20 minutes ) ; nucleated cells have the ability to resynthesize acetylated P36551 isozymes within a few hours , while platelets do not . P36551 -independent mechanisms of aspirin have been suggested to explain its chemopreventive effects but this concept remains to be demonstrated in vivo at clinical doses . Short-term/low-dose aspirin-induced duodenal erosions are not dependent on Helicobacter pylori infection , cyclooxygenase expression and prostaglandin E2 levels . OBJECTIVE : The mechanisms of interaction between Helicobacter pylori infection and low-dose aspirin in the induction of duodenal erosions are not fully understood . The aim of this study was to investigate the effects of low-dose aspirin on the induction of duodenal erosions , the expression of cyclooxygenases and prostaglandin ( PG ) -E(2) levels in healthy subjects according to their H. pylori status . MATERIAL AND METHODS : Twenty healthy volunteers ( H. pylori-negative n=10 , H. pylori-positive n=10 ) received 100 mg aspirin/day for 1 week . During esophagogastroduodenoscopy , duodenal biopsies were taken before and at days 1 , 3 , and 7 of medication . P23219 and -2 expressions were analyzed by reverse transcriptase-polymerase chain reaction ( RT-PCR ) , and immunohistochemistry ; mucosal PGE(2) levels were determined by ELISA . Three months after successful eradication of infection , nine H. pylori-positive subjects repeated the protocol . RESULTS : DB00945 -induced duodenal erosions occurred independently of whether H. pylori infection was present or not . There was no difference in duodenal P23219 and P35354 expression among the groups and expression was not affected by aspirin . Basal duodenal PGE(2) levels were similar among the different groups ( H. pylori-negative 4.3+/-4.2 , H. pylori-positive 5.2+/-1.3 , following H. pylori eradication 5.2+/-1.4 ng/microg protein ) and were not affected by low-dose aspirin . CONCLUSIONS : In healthy subjects , low-dose aspirin-induced duodenal erosions are not influenced by H. pylori status . Low-dose aspirin medication for one week does not affect either cyclooxygenase expression or duodenal PGE(2) levels and therefore is likely to induce duodenal damage mainly through topical toxicity . DB00945 reduces the prothrombotic activity of P02741 . AIM : P02741 ( CRP ) is a risk marker and a potential modulator of vascular disease . Previous studies support a prothrombotic activity of CRP , with impaired thromboregulation . The present study examined the antithrombotic effect of aspirin in mice transgenic for human CRP ( CRPtg mice ) . Mechanistic investigations further elucidated the effect of CRP on prostanoid metabolism in vivo and in vitro . METHODS AND RESULTS : Administration of aspirin ( 30 mg kg(-1) day(-1) ) to CRPtg mice slowed the accelerated thrombosis after photochemical injury to the carotid ( 99 +/- 32 vs. 45 +/- 24 min and 75 +/- 23 vs. 82 +/- 26 min in wild-type mice vs. CRPtg mice , without and following aspirin treatment , respectively ) . Vascular injury modulated the expression of key pathways in prostanoid metabolism differently in CRPtg mice and wild-type mice . Suppression of cyclo-oxygenase 2 ( P35354 ) -derived metabolism with suppression of prostaglandin I2 ( DB01240 ) synthase and DB01240 metabolism was recorded in the injured artery with increased thromboxane receptor expression . DB00945 therapy reduced the difference in DB01240 biosynthesis between CRPtg mice and wild-type mice . In vitro studies in human-derived cells further supported these findings . Incubation of human umbilical vein endothelial cells ( HUVECs ) with human recombinant CRP ( 5 microg mL(-1) ) suppressed DB01240 synthase expression and significantly increased thromboxane receptor levels . Incubation of smooth muscle cells with CRP did not affect prostanoid expression . CONCLUSIONS : CRP modulates prostanoid metabolism to favor vascular occlusion . Elevated CRP levels might predispose to the cardiovascular hazard conferred by selective P35354 inhibitors , and the risk mediated by CRP may be limited by aspirin . 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 . 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 . DB00945 -induced nuclear translocation of NFkappaB and apoptosis in colorectal cancer is independent of p53 status and DNA mismatch repair proficiency . Substantial evidence indicates nonsteroidal anti-inflammatory drugs ( NSAIDs ) protect against colorectal cancer ( CRC ) . However , the molecular basis for this anti-tumour activity has not been fully elucidated . We previously reported that aspirin induces signal-specific P25963 degradation followed by NFkappaB nuclear translocation in CRC cells , and that this mechanism contributes substantially to aspirin-induced apoptosis . We have also reported the relative specificity of this aspirin-induced NFkappaB-dependent apoptotic effect for CRC cells , in comparison to other cancer cell types . It is now important to establish whether there is heterogeneity within CRC , with respect to the effects of aspirin on the NFkappaB pathway and apoptosis . p53 signalling and DNA mismatch repair ( P22897 ) are known to be deranged in CRC and have been reported as potential molecular targets for the anti-tumour activity of NSAIDs . Furthermore , both p53 and P22897 dysfunction have been shown to confer resistance to chemotherapeutic agents . Here , we set out to determine the p53 and hMLH1 dependency of the effects of aspirin on NFkappaB signalling and apoptosis in CRC . We specifically compared the effects of aspirin treatment on cell viability , apoptosis and NFkappaB signalling in an HCT-116 CRC cell line with the p53 gene homozygously disrupted ( HCT-116(p53-/-) ) and an HCT-116 cell line rendered P22897 proficient by chromosomal transfer ( HCT-116(+ch3) ) , to the parental HCT-116 CRC cell line . We found that aspirin treatment induced apoptosis following P25963 degradation , NFkappaB nuclear translocation and repression of NFkappaB-driven transcription , irrespective of p53 and DNA P22897 status . These findings are relevant for design of both novel chemopreventative agents and chemoprevention trials in CRC . P15692 , P17948 , P35968 and colorectal cancer : assessment of disease risk , tumor molecular phenotype , and survival . Angiogenesis is essential for tumor progression . Vascular endothelial growth factor ( P15692 ) and its receptors 1 ( P17948 ) and 2 ( P35968 ) , have been identified as major mediators of this process . We hypothesized that genetic variation in P17948 ( 38 SNPs ) , P35968 ( 22 SNPS ) , and P15692 ( 11 SNPs ) would be associated with colon and rectal cancer development and survival . Data from a case-control study of 1555 colon cancer cases and 1956 controls and 754 rectal cancer cases and 959 controls were used . An adaptive rank truncation product ( ARTP ) , based on 10,000 permutations , was used to determine the statistical significance of the candidate genes and angiogenesis pathway . Based on ARTP results , P17948 was significantly associated with risk of colon cancer ( P(ARTP) = 0.045 ) and P15692 was significantly associated with rectal cancer ( P(ARTP) = 0.036 ) . After stratifying by tumor molecular subtype , SNP associations observed for colon cancer were : P15692 rs2010963 with CIMP+ colon tumors ; P17948 rs4771249 and rs7987649 with P04637 ; P17948 rs3751397 , rs7337610 , rs7987649 , and rs9513008 and P35968 rs10020464 , rs11941492 , and rs12498529 with MSI+ and CIMP+/ P01116 -mutated tumors . P17948 rs2296189 and rs600640 were associated with CIMP+ rectal tumors and P17948 rs7983774 was associated with P04637 -mutated rectal tumors . Four SNPs in P17948 were associated with colon cancer survival while three SNPs in P35968 were associated with survival after diagnosis with rectal cancer . DB00945 /NSAID use , smoking cigarettes , and BMI modified the associations . These findings suggest the importance of inflammation and angiogenesis in the etiology of colorectal cancer and that genetic and lifestyle factors may be targets for modulating disease risk . 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 aspirin and nonsteroidal anti-inflammatory drugs on prostaglandins . Cyclic prostanoids play important physiologic roles in inflammation and maintaining normal function of several organ systems . Prostaglandin production requires the conversion of arachidonate to the intermediate prostaglandin H2 , by the 2-step cyclo-oxygenation and peroxidation catalyzed by the enzyme cyclo-oxygenase ( also called prostaglandin H synthase ) . DB00945 and nonsteroidal anti-inflammatory drugs ( NSAIDs ) block the production of cyclic prostanoids by binding in different ways to this enzyme and blocking the active site . This results in decreased inflammation , but it can also produce side effects in the gastrointestinal tract , kidney , and platelets . Recent data demonstrate that there are 2 isoforms of the cyclo-oxygenase enzyme , called P23219 and P35354 . These isoforms are similar in size , substrate specificity , and kinetics , but vary in their expression and distribution . Normal physiologic functions appear to be maintained by P23219 , while P35354 appears to mediate the inflammatory response . Nonsteroidal drugs with selective inhibitory activity on the P35354 isoform should theoretically decrease inflammation while maintaining normal physiologic prostaglandin levels . Current NSAIDs are not selective enough to confirm this , but newer , more selective inhibitors of P35354 may answer this important question . 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 ) . 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 . Adulthood nicotine treatment alleviates behavioural impairments in rats neonatally treated with quinpirole : possible roles of acetylcholine function and neurotrophic factor expression . Increases in dopamine D(2) receptor sensitivity are known to be common in drug abuse and neurological disorders . Past data from this laboratory have shown that long-term increases in D(2) sensitivity can be produced by quinpirole treatment ( a D(2)/D(3) agonist ) during early development . The present investigation was designed to test the hypothesis that nicotine administration in adulthood would reduce both cognitive and skilled reaching impairments produced by increases in D(2) sensitivity . Female Sprague-Dawley rats were treated with quinpirole ( 1 mg/kg ) or saline from postnatal day 1 ( PD 1 ) to PD 21 . Beginning in adulthood ( PD 61 ) , rats were treated with nicotine ( 0.3 mg/kg free base ) or saline twice daily for 14 consecutive days before behavioural testing commenced . Animals neonatally treated with quinpirole demonstrated performance deficits on the Morris water task and a skilled reaching task compared to controls . Deficits on both tasks were completely alleviated by adulthood nicotine treatment . Animals neonatally treated with quinpirole demonstrated a significant 36 % decrease of P28329 in the hippocampus compared to saline controls that was partially eliminated by nicotine . Additionally , neonatal quinpirole produced a significant decrease in hippocampal P01138 content compared to controls , however , nicotine failed to alleviate this decrease in P01138 . The results of this investigation demonstrate that long-term increases in dopamine D(2) receptor sensitivity produce significant decreases in hippocampal cholinergic and P01138 expression that may result in cognitive impairment . DB00184 alleviates both cognitive and skilled reaching impairments caused by increases in D(2) sensitivity , but the mechanism through which nicotine is acting is currently unknown . 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 . 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 . P15692 expression and the effect of NSAIDs on ascites cell proliferation in the hen model of ovarian cancer . OBJECTIVES : We aimed to determine the expression of vascular endothelial growth factor ( P15692 ) and the effect of nonsteroidal anti-inflammatory drugs ( NSAIDs ) on the proliferation of cells isolated from ascites in the hen model of ovarian cancer . METHODS : Ovarian tumor and normal ovary were collected from hens and ascites cells were isolated from hens with ovarian cancer . Quantitative real-time PCR was used to quantify mRNA expression . Immunohistochemical and/or Western blot analyses were used to localize protein expression in ovarian tumors , normal ovaries , and ascites cells . Cells were treated with a nonspecific , P23219 -specific , or P35354 -specific NSAID and proliferation was determined . RESULTS : P15692 mRNA was increased in ascites cells and there was a trend for a correlation between P15692 mRNA in ascites cells and ascites volume . P15692 protein was localized to theca cells of normal ovaries , in glandular areas of tumors , and to the cytoplasm of ascites cells . DB00945 and a P23219 -specific inhibitor decreased the proliferation of ascites cells , whereas a P35354 -specific inhibitor did not . CONCLUSIONS : P15692 may play a role in ovarian cancer progression in the hen and the proliferation of ascites cells can be decreased by targeting the P23219 but not P35354 pathway . 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 . Effect of short-term , low dose aspirin supplementation on the activation of pro-inflammatory NF-kappaB in aged rats . The basis of our recently proposed " molecular inflammation theory of aging " is that activated inflammatory transcription factors , including versatile NF-kappaB , occur widespread in the organism during aging . NF-kappaB plays a key role in pro-inflammatory gene expression , such as cyclooxygenase ( P36551 ) . DB00945 is one of the most commonly used non-steroidal anti-inflammatory drugs because of its ability to inhibit P36551 activity . Thus , in the present study , we investigated the effect of short-term , low dose aspirin intake on the modulation of pro-inflammatory NF-kappaB activation in old rats . To conduct the study , 24-month-old Fischer 344 rats were supplemented with low dose aspirin ( 0.015 % ) for 10 days . Biochemical analyses showed suppressed reactive species ( RS ) and P35354 activity . The data also showed that NF-kappaB activation and its associated gene expressions , such as P35354 , P35228 , P19320 and P05362 , were all suppressed by the low dose aspirin supplementation through the inhibition of phosphorylation and degradation of P25963 via the NIK/IKK pathway . Our molecular exploration further revealed that aspirin 's suppressive action of NF-kappaB was mediated by its ability to inhibit the nuclear translocation of cytosolic thioredoxin and redox factor-1 . These findings showed for the first time that in aged rat short-term low dose dietary aspirin feeding modulates the molecular signal transduction involved in the inflammatory process . P35354 is a neuronal target gene of NF-kappaB . BACKGROUND : NF-kappaB is implicated in gene regulation involved in neuronal survival , inflammmatory response and cancer . There are relatively few neuronal target genes of NF-kappaB characterized . RESULTS : We have identified the neuronal cyclooxygenase-2 ( P35354 ) as a NF-kappaB target gene . In organotypic hippocampal slice cultures constitutive NF-kappaB activity was detected , which was correlated with high anti- P35354 immunoreactivity . DB00945 a frequently used painkiller inhibits neuronal NF-kappaB activity in organotypic cultures resulting in a strong inhibition of the NF-kappaB target gene P35354 . Based on these findings , the transcriptional regulation of P35354 by NF-kappaB was investigated . Transient transfections showed a significant increase of P35354 promoter activity upon stimulation with PMA , an effect which could be obtained also by cotransfection of the NF-kappaB subunits p65 and p50 . In the murine neuroblastoma cell line NB-4 , which is characterized by constitutive NF-kappaB activity , P35354 promoter activity could not be further increased with PMA or P01375 . Constitutive promoter activity could be repressed upon cotransfection of the inhibitory subunit IkappaB-alpha . EMSA and mutational analysis conferred the regulatory NF-kappaB activity to the promoter distal kappaB-site in the human P35354 promoter . CONCLUSIONS : NF-kappaB regulates neuronal P35354 gene expression , and acts as an upstream target of DB00945 . This extends DB00945 's mode of action from a covalent modification of P35354 to the upstream regulation of P35354 gene expression in neurons . 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 . [ DB00945 ] . Acetylsalicylic acid ( aspirin ) was the first synthetized drug 100 years ago . Whereas other drugs have disappeared a long time ago , aspirin has stood the test of time and is one of the most important drugs at the turn of this century . Initially it was used for its analgesic , antipyretic and antiphlogistic properties . They are due to the acetylation of the cyclooxygenase and hence an inhibition of prostaglandin synthesis . At higher doses aspirin inhibits interleukin expression induced by nuclear factor-kappa B . DB00945 is the prototype of nonsteroidal antiinflammatory drugs , which are currently challenged by the development of specific inhibitors of cyclooxygenase isoform 2 ( P35354 ) , which must , however , first prove their efficacy and safety in clinical trials . The inhibition of platelet aggregation by aspirin has only been recognized in the second half of the century . It is due to an inhibition of thromboxane A2 synthesis because of an irreversible cyclooxygenase blockade in platelets . DB00945 has been found to reduce the incidence and death rate of cardiovascular and cerebrovascular events and is nowadays the cornerstone of any secondary prevention in vascular diseases . Newer antiaggregatory agents such as ticlopidine , clopidogrel or IIb/IIIa-blockers have been developed . Most often they are used in conjunction with aspirin and their place has yet to be defined . New modes of action of aspirin continue to be found . Recent examples are an improvement of endothelial dysfunction or a reduced incidence of colorectal cancers . It is therefore likely that aspirin will continue to be a very useful and cheap drug for a very large population and will meet the interest of researchers for many more decades . Non-small cell lung cancer cycloxygenase activity and proliferation are inhibited by non-steroidal antiinflammatory drugs . The effects of non-steroidal antiinflammatory drugs ( NSAIDs ) on non-small cell lung cancer ( NSCLC ) were investigated . Arachidonic acid ( AA ) was metabolized to prostaglandin E2 ( DB00917 ) in NSCLC cells . NSAIDs such as aspirin or indomethacin reduced DB00917 levels in NCI-H157 and H1264 cells , and the decrease caused by DB00917 was reversed by epidermal growth factor ( P01133 ) . By RT-PCR , both cyclooxygenase ( P36551 ) -1 and P35354 mRNAs are detected in NCI-H157 and H1264 cells . By Northern analysis , P35354 mRNA was induced by P01133 and phorbol ester . By immunocytochemistry , P23219 and P35354 enzymes were localized to NSCLC tumors . DB00945 , indomethacin and ibuprofen decreased NSCLC growth in vitro . DB00945 and indomethacin inhibited proliferation of NSCLC xenografts in nude mice . These data suggest that P36551 enzymes may be important regulatory components of NSCLC . 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 . DB00945 and nonsteroidal anti-inflammatory drug hypersensitivity . Acetylsalicylic acid ( ASA ) or aspirin and nonsteroidal anti-inflammatory drug ( NSAID ) sensitivities encompass a diverse group of both pharmacological and hypersensitivity reactions . Conventionally , hypersensitivities include aspirin-exacerbated respiratory disease ( AERD ) , ASA-induced urticaria , and anaphylaxis . With an increasing prevalence of coronary artery disease in an aging population , aspirin continues to play a significant role in cardiac prophylaxis in a large patient population . Invariably , the clinician will encounter patients with clear indications for aspirin therapy but a history of aspirin sensitivity . Although protocols have been established for aspirin challenge and desensitization , it is not always an efficacious or safe procedure . This article reviews the different classifications of ASA/NSAIDs hypersensitivities to better guide the clinician in dealing with this patient population . History of crossrelativities between multiple NSAIDs implies a non-IgE-mediated process . Similarly , a history of monosensitivity to one NSAID implies an IgE-mediated process , although specific antibodies are often elusive . Despite the name , AERD can potentially be exacerbated by all cyclooxygenase ( P36551 ) inhibitors based on dose-dependent inhibition of P23219 . DB00945 desensitization can be achieved to improve both upper and lower respiratory symptoms for most patient with AERD . DB00945 desensitization can usually be achieved for those in need of the antiplatelet effects of aspirin , with the exception of those with aspirin-induced urticaria and baseline chronic urticaria . However , desensitization should only be attempted in those with stable coronary artery disease because the process of desensitization carries the inherent risk of anaphylaxis/anaphylactoid reaction , which may further increase cardiac demand and bring about ischemic injury . Therefore , desensitization is reserved until coronary artery disease is stabilized . Assessment of common nonsteroidal anti-inflammatory medications by whole blood aggregometry : a clinical evaluation for the perioperative setting . OBJECTIVE : To help define the perioperative risk related to commonly used non-aspirin NSAIDs with whole blood platelet aggregometry . METHODS : Twelve healthy volunteers were recruited . Two cyclooxygenase ( P36551 ) -1 inhibitors ( ibuprofen and naproxen ) and two P35354 inhibitors ( meloxicam and celecoxib ) were administered , and daily whole blood platelet aggregometry studies were obtained until studies showed no platelet inhibition . DB00945 was studied at the conclusion of the study . RESULTS : Ibuprofen had no inhibitory effect on platelet aggregation in all women and no inhibitory effect in 83 % of men at 24 hours . All platelet function had returned to normal at 48 hours . The inhibitory effect of naproxen on platelets was absent at 48 hours in 83 % of the women and 50 % of men . By 72 hours all platelet studies had returned to normal . Meloxicam and celecoxib did not cause any overall inhibitory effect on platelet aggregation . CONCLUSIONS : Ibuprofen and naproxen have a mild inhibitory effect on platelet aggregation compared with aspirin and this effect is undetectable by 48 hours and 72 hours , respectively . Meloxicam and celecoxib show essentially no inhibitory effect on platelet aggregation . These findings suggest that there is little bleeding risk related to platelet aggregation at 24 hours in patients who take P35354 inhibitors and at 72 hours for those who take P23219 inhibitor medications . Cyclooxygenase inhibition and thrombogenicity . Cyclooxygenase ( P36551 ) -1 and P35354 catalyze the formation of prothrombotic and antithrombotic eicosanoids , respectively . DB00945 , conventional nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and P35354 -specific inhibitors exhibit different patterns of inhibition of P23219 -mediated thromboxane biosynthesis and P35354 -mediated prostacyclin biosynthesis . The relationship between the pharmacologic inhibition of these vasoactive eicosanoids and the thromboprophylaxis or thrombogenicity exhibited by different therapeutic agents is currently unclear . Future studies are needed to assess the antithrombotic properties of commonly used NSAIDs , the hypothetical thrombogenicity of P35354 -specific inhibitors in high-risk patients , the need for concomitant aspirin with selective versus nonselective P36551 inhibitors , and the antiplatelet and gastric toxicity of the aspirin/ P35354 -specific inhibitor combination in comparison with the aspirin/conventional NSAID combination . 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 . 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 . Vane 's discovery of the mechanism of action of aspirin changed our understanding of its clinical pharmacology . DB00945 exerts its analgesic , antipyretic and anti-inflammatory actions by inhibiting the enzyme cyclooxygenase and thus preventing the formation and release of prostaglandins . The elucidation by John Vane of the mechanism of action of aspirin in 1971 was followed twenty years later by the discovery of a second cyclooxygenase enzyme , P35354 and the rapid development of selective inhibitors of this enzyme . The P35354 inhibitors are potent anti-inflammatory drugs without the damaging side effects on the stomach mucosa of the non-selective aspirin-like inhibitors . More recently , two enzymes have been identified inhibition of which may explain the mechanism of action of paracetamol . These are a putative cyclooxygenase-3 which is a variant of cyclooxygenase-1 and derives from the same gene , and a P35354 variant , induced with diclofenac , which may be involved in the resolution of inflammation . Expression of genes associated with bone resorption is increased and bone formation is decreased in mice fed a high-fat diet . A high-fat diet ( HFD ) leads to an increased risk of osteoporosis-related fractures , but the molecular mechanisms for its effects on bone metabolism have rarely been addressed . The present study investigated the possible molecular mechanisms for the dyslipidemic HFD-induced bone loss through comparing femoral gene expression profiles in HFD-fed mice versus the normal diet-fed mice during the growth stage . We used Affymetrix 430A Gene Chips to identify the significant changes in expression of the genes involved in bone metabolism , lipid metabolism , and the related signal transduction pathways . Quantitative RT-PCR was carried out on some significant genes for corroboration of the microarray results . At the conclusion of the 12-week feeding , the down-regulation of most of the genes related to bone formation and the up-regulation of most of the genes related to bone resorption were observed in the HFD-fed mice , consistent with the changes in plasma bone metabolic biomarkers . Together , the HFD induced a decrease in the majority of the adipogenesis- , lipid biosynthesis- , and fatty acid oxidation-related gene expression , such as PPARg and P02649 . Furthermore , some genes engaged in the related signal transduction pathway were strongly regulated at the transcript level , including P22692 , TGFbR1 , IL-17a , P05112 , and P04637 . These results indicate that an HFD may induce inhibitory bone formation and enhanced bone resorption , thus causing adverse bone status . DB00945 -responsive , migraine-like transient cerebral and ocular ischemic attacks and erythromelalgia in O60674 -positive essential thrombocythemia and polycythemia vera . Migraine-like cerebral transient ischemic attacks ( MIAs ) and ocular ischemic manifestations were the main presenting features in 10 O60674 (V617F)-positive patients studied , with essential thrombocythemia ( ET ) in 6 and polycythemia vera ( PV ) in 4 . Symptoms varied and included cerebral ischemic attacks , mental concentration disturbances followed by throbbing headaches , nausea , vomiting , syncope or even seizures . MIAs were frequently preceded or followed by ocular ischemic events of blurred vision , scotomas , transient flashing of the eyes , and sudden transient partial blindness preceded or followed erythromelalgia in the toes or fingers . The time lapse between the first symptoms of aspirin-responsive MIAs and the diagnosis of ET in 5 patients ranged from 4 to 12 years . At the time of erythromelalgia and MIAs , shortened platelet survival , an increase in the levels of the platelet activation markers β-thromboglobulin and platelet factor 4 and also in urinary thromboxane B2 were clearly indicative of the spontaneous in vivo platelet activation of constitutively O60674 (V617F)-activated thrombocythemic platelets . DB00945 relieves the peripheral , cerebral and ocular ischemic disturbances by irreversible inhibition of platelet cyclo-oxygenase ( P23219 ) activity and aggregation ex vivo . Vitamin K antagonist , dipyridamole , ticlopidine , sulfinpyrazone and sodium salicylate have no effect on platelet P23219 activity and are ineffective in the treatment of thrombocythemia-specific manifestations of erythromelalgia and atypical MIAs . If not treated with aspirin , ET and PV patients are at a high risk of major arterial thrombosis including stroke , myocardial infarction and digital gangrene . 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 . Regulation of Cu/Zn-superoxide dismutase expression via the phosphatidylinositol 3 kinase/Akt pathway and nuclear factor-kappaB . Aerobic cells adjust the expression of antioxidant enzymes to maintain reactive oxygen species within tolerable levels . In addition , phosphatidylinositol 3 kinase ( PI3K ) and its downstream protein kinase effector Akt adapt cells to survive in the presence of oxidative stress . Here we provide evidence for an association between these two defense systems via transcriptional regulation of Cu/Zn-superoxide dismutase ( Cu/Zn-SOD ) . PC12 pheochromocytoma cells expressing active Akt1 exhibit lower ROS levels in response to hydrogen peroxide , as determined with the superoxide-sensitive probe hydroethidine . Transfection of constitutive or 4-hydroxytamoxifen-inducible versions of Akt1 results in higher messenger RNA and protein levels of Cu/Zn-SOD . Luciferase reporter constructs , carrying different length fragments of the human sod1 gene promoter , have identified a region between -552 and -355 that is targeted by PI3K and Akt and that contains a putative site of regulation by nuclear factor-kappaB ( NF-kappaB ) . Nerve growth factor ( P01138 ) and Akt augment the transactivating activity and produce higher nuclear levels of p65-NF-kappaB . Electrophoretic mobility shift assays indicate that the putative NF-kappaB regulatory sequence binds p65-NF-kappaB more efficiently in nuclear extracts from these cells . A dominant-negative mutant of P25963 further demonstrates that the PI3K/Akt axis targets the sod1 promoter at the level of the newly characterized NF-kappaB site . These results illustrate a new mechanism by which the PI3K/Akt pathway protects cells against oxidative stress , involving the upregulation of Cu/Zn-SOD gene expression , and the results identify NF-kappaB as a key mediator in the regulation of this gene . 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 . 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 . DB00945 analogues as dual cyclooxygenase-2/ P09917 inhibitors : synthesis , nitric oxide release , molecular modeling , and biological evaluation as anti-inflammatory agents . Analogues of aspirin were synthesized through an efficient one-step reaction in which the carboxyl group was replaced by an ethyl ester , and/or the acetoxy group was replaced by an N-substituted sulfonamide ( SO(2)NHOR(2):R(2) =H , Me , CH(2)Ph ) pharmacophore . These analogues were designed for evaluation as dual cyclooxygenase-2 ( P35354 ) and P09917 ( 5- P28300 ) inhibitors . In vitro P23219 / P35354 isozyme inhibition studies identified compounds 11 ( CO(2) H , SO(2)NHOH ) , 12 ( CO(2)H , SO(2)NHOCH(2)Ph ) , and 16 ( CO(2)Et , SO(2)NHOH ) as highly potent and selective P35354 inhibitors ( IC(50) range : 0.07-0.7 μM ) , which exhibited appreciable in vivo anti-inflammatory activity ( ED(50) range : 23.1-31.4 mg kg(-1) ) . Moreover , compounds 11 ( IC(50) =0.2 μM ) and 16 ( IC(50) =0.3 μM ) , with a sulfohydroxamic acid ( SO(2)NHOH ) moiety showed potent 5- P28300 inhibitory activity . Furthermore , the SO(2)NHOH moiety present in compounds 11 and 16 was found to be a good nitric oxide ( NO ) donor upon incubation in phosphate buffer at pH 7.4 . Molecular docking studies in the active binding site of P35354 and 5- P28300 provided complementary theoretical support for the experimental biological structure-activity data acquired . 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 . 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 . DB00945 : new cardiovascular uses for an old drug . Inhibition of TXA2-dependent platelet function by aspirin may lead to prevention of thrombosis as well as to excess bleeding . The balance between the two depends critically on the absolute thrombotic versus hemorrhagic risk of the patient . As the risk of experiencing a major vascular event increases , so does the absolute benefit of antiplatelet prophylaxis with aspirin [ Figure-see text ] . The antithrombotic effect of aspirin does not appear to be dose related over a wide range of daily doses ( 30 to 1,300 mg ) , an observation consistent with saturability of platelet P23219 inhibition by aspirin at very low doses . In contrast , GI toxicity of the drug does appear to be dose related , consistent with dose- and dosing interval-dependent inhibition of P23219 activity in the nucleated lining cells of the GI mucosa . Thus , aspirin once daily is recommended in all clinical conditions where antiplatelet therapy is effective . Because of safety considerations , physicians are encouraged to use the lowest dose of aspirin shown effective in each clinical setting [ Table-see text ] . 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 . 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 ) 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 . The epidemiology of prostate cancer -- with a focus on nonsteroidal anti-inflammatory drugs . DB00945 and other NSAIDs have a potential role in the primary and secondary prevention of many common diseases associated with aging , including the top two causes of mortality in the United States-cardiovascular disease and cancer . These agents may be beneficial in the management of Alzheimer 's disease,other forms of dementia , and Parkinson's. disease . Because men with prostate cancer or precancer are likely to present with coexisting conditions that would be affected by systemic aspirin , NSAID , or other P35354 inhibitor therapies , it is important to consider any possible preventive studies or future clinical recommendations of aspirin or NSAIDs for prostate cancer within the context of these comorbid conditions . DB00945 or nonaspirin NSAIDs may be appropriate prevention therapy for patients at high risk of prostate cancer , myocardial infarction , Parkinson 's disease , Alzheimer 's disease , lung cancer , or colorectal cancer , but low risk for gastrointestinal complications or stroke . Further quantitative comparative studies of the risks and benefits of these common comorbidities in older Americans , with special attention to dose and duration parameters , are warranted . 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 . 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 . 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 . 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 .

MH_train_1152

MH_train_1152

MH_train_1152

interacts_with DB01045?

multiple_choice

DNA damage and S phase arrest induced by Ochratoxin A in human embryonic kidney cells ( P29320 293 ) . Ochratoxin A ( OTA ) is a ubiquitous mycotoxin with potential nephrotoxic , hepatotoxic and immunotoxic effects . The mechanisms underlying the nephrotoxicity of OTA remain obscure . To investigate DNA damage and the changes of the cell cycle distribution induced by OTA , human embryonic kidney cells ( P29320 293 cells ) were incubated with various concentrations of OTA for 24h in vitro . The results indicated that OTA treatment led to the production of reactive oxygen species ( ROS ) and to a decrease of the mitochondrial membrane potential ( ΔΨm ) . OTA-induced DNA damage in P29320 293 cells was evidenced by DNA comet tails formation and increased expression of γ- P16104 . In addition , OTA could induce cell cycle arrest at the S phase in P29320 293 cells . The expression of key cell cycle regulatory factors that were critical to the S phase , including cyclin A2 , cyclin E1 , and P24941 , were further detected . The expression of cyclin A2 , cyclin E1 , and P24941 were significantly decreased by OTA treatment at both the mRNA and protein levels . The apoptosis of P29320 293 cells after OTA treatment was observed using Hoechst 33342 staining . The results confirmed that OTA did induce apoptosis in P29320 293 cells . In conclusion , our results provided new insights into the molecular mechanisms by which OTA might promote nephrotoxicity . Effect of P15941 siRNA on drug resistance of gastric cancer cells to trastuzumab . DB00072 is the first molecular targeting drug to increase the overall survival rate in advanced gastric cancer . However , it has also been found that a high intrinsic or primary trastuzumab resistance exists in some proportion of gastric cancer patients . In order to explore the mechanism of resistance to trastuzumab , firstly we investigated the expression of P15941 ( membrane-type mucin 1 ) in gastric cancer cells and its relationship with drug-resistance . Then using gene-silencing , we transfected a siRNA of P15941 into drug-resistant cells . The results showed the MKN45 gastric cell line to be resistant to trastuzumab , mRNA and protein expression of P15941 being significantly upregulated . After transfection of P15941 siRNA , protein expression of P15941 in MKN45cells was significantly reduced . Compared with the junk transfection and blank control groups , the sensitivity to trastuzumab under P15941 siRNA conditions was significantly increased . These results imply that P04626 -positive gastric cancer cell MKN45 is resistant to trastuzumab and this resistance can be cancelled by silencing expression of the P15941 gene . 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 . 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 . An intricate network of conserved DNA upstream motifs and associated transcription factors regulate the expression of uromodulin gene . PURPOSE : P07911 is a kidney specific glycoprotein whose expression can modulate kidney homeostasis . However , the set of sequence specific transcription factors that regulate the uromodulin gene P07911 and their upstream binding locations are not well characterized . We built a high resolution map of its transcriptional regulation . MATERIALS AND METHODS : We applied in silico phylogenetic footprinting on the upstream regulatory regions of a diverse set of human P07911 orthologs to identify conserved binding motifs and corresponding position specific weight matrices . We further analyzed the predicted binding motifs by motif comparison , which identified transcription factors likely to bind these discovered motifs . Predicted transcription factors were then integrated with experimentally known protein-protein interactions available from public databases and tissue specific expression resources to delineate important regulators controlling P07911 expression . RESULTS : Analysis allowed the identification of a reliable set of binding motifs in the upstream regulatory regions of P07911 to build a high confidence compendium of transcription factors that could bind these motifs , such as P23771 , P35680 , SP1 , P84022 , Q13950 and O43474 . ENCODE deoxyribonuclease I hypersensitivity sites in the P07911 upstream region of the mouse kidney confirmed that some of these binding motifs were open to binding by predicted transcription factors . The transcription factor-transcription factor network revealed several highly connected transcription factors , such as SP1 , Q02447 , P04637 , P14859 , P10826 , P10276 and P19793 , as well as the likely protein complexes formed between them . Expression levels of these transcription factors in the kidney suggest their central role in controlling P07911 expression . CONCLUSIONS : Our findings will form a map for understanding the regulation of uromodulin expression in health and disease . Effect of lipopolysaccharide on the xenobiotic-induced expression and activity of hepatic cytochrome P450 in mice . Infection-associated inflammation can alter the expression levels and functions of cytochrome P450s ( CYPs ) . Cyp gene expression is regulated by the activation of several nuclear receptors , including pregnane X receptor ( O75469 ) , constitutive androstane receptor ( CAR ) , and aryl hydrocarbon receptor ( P35869 ) . These receptors can be activated by xenobiotics , including medicines . Here , to study the xenobiotic-induced fluctuations in CYP during inflammation , we examined the effect of lipopolysaccharide ( LPS ) treatment on the level of mRNAs encoding hepatic CYPs induced by xenobiotic-activated nuclear receptors , in mice . Both the mRNA induction of Cyp genes and the metabolic activities of CYP proteins were examined . LPS treatment caused a significant decrease in the induced expression of the mRNAs for Cyp3a11 , 2c29 , 2c55 , and 1a2 , but not for Cyp2b10 . To assess the CYP enzymatic activities , CYP3A-mediated testosterone 6β-hydroxylation and the intrinsic clearance ( CL(int) ) of nifedipine in liver microsomes were measured in mice treated with the xenobiotic pregnenolone-16alpha-carbonitrile ( Q15149 ) with or without LPS administration . Both assays revealed that the CYP3A activity , which was induced by Q15149 , declined significantly after LPS treatment , and this decline correlated with the Cyp3a11 mRNA level . In addition , we found that the mRNAs for interleukin ( IL ) -1β and tumor necrosis factor ( P01375 ) α were increased after treatment with LPS plus xenobiotics . Our findings demonstrated that LPS treatment reduces the O75469 - and P35869 -mediated , and possibly CAR-mediated Cyp gene expression and further suggest that these decreases are dependent on inflammatory cytokines in the liver . Molecular dissection of dimethylnitrosamine ( O15061 ) -induced hepatotoxicity by mRNA differential display . Differential display reverse transcription-polymerase chain reaction ( DDRT-PCR ) was used to catalogue altered hepatic transcript expression during dimethylnitrosamine ( O15061 ) exposure in vivo . Mice were administered O15061 ( 1.5 or 5 mg/kg ) or vehicle ( phosphate-buffered saline ) i.p. once daily for up to 7 days , and livers were collected 6 h post-injection . Total RNA was reverse transcribed and cDNA subsets were selectively amplified by PCR . DDRT-PCR products were fractionated on denaturing polyacrylamide gels , and differentially expressed bands were excised , reamplified , and subsequently cloned into a plasmid vector . This study identified 23 cDNAs that were induced and 25 cDNAs that were suppressed during O15061 exposure . Altered expression during O15061 exposure for cDNA clones was confirmed by Northern blotting , RNase protection , or in situ hybridization analyses . DNA sequence information indicated that four cDNAs suppressed during O15061 exposure encode cytochrome P450 isoenzyme-cholesterol 7 alpha-hydroxylase ( P22680 ) , a monokine , a myeloid cell differentiation protein , and mouse major urinary protein ( MUP ) . We further observed a O15061 -induced increase in transcripts for complement factor 3 ( P01024 ) and serum amyloid A ( P0DJI8 ) . In contrast , the remaining differentially expressed transcripts detected by DDRT-PCR during O15061 exposure demonstrated no similarity to sequences present in Genbank , suggesting that they may encode previously unreported gene products . In situ hybridization showed MUP transcripts to be expressed by hepatic centrilobular areas that undergo necrosis during subchronic O15061 exposure . Thus , the utilization of DDRT-PCR has identified several differentially expressed hepatic mRNAs associated with various doses and stages of O15061 exposure . 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 . 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 . 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 . 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 . A 4-week intrathecal toxicity and pharmacokinetic study with trastuzumab in cynomolgus monkeys . DB00072 is indicated for the treatment of patients with breast cancer overexpressing human epidermal growth factor 2 ( P04626 ) . Women with P04626 -positive tumors have an increased risk of brain metastases . The blood-brain barrier and blood-cerebrospinal fluid ( P04141 ) barrier may prevent trastuzumab from reaching appropriate concentrations in the brain and P04141 following standard intravenous administration . To evaluate the potential of effects on the central nervous system , a 4-week toxicology study with weekly intrathecal administration of trastuzumab was performed in cynomolgus monkeys at doses of 0 , 3 , or 15 mg . No trastuzumab-related effects on body weight , clinical signs , neurological function , clinical pathology , or anatomic pathology were noted . The applied doses and P04141 concentrations achieved in the current study exceeded those reported in patients after intrathecal administration . The results support future studies for further evaluation of intrathecal application of trastuzumab in patients with brain metastases in P04626 -positive breast cancer . An LXR agonist promotes glioblastoma cell death through inhibition of an P00533 /AKT/ P36956 / P01130 -dependent pathway . Glioblastoma ( GBM ) is the most common malignant primary brain tumor of adults and one of the most lethal of all cancers . P00533 ( P00533 ) mutations ( EGFRvIII ) and phosphoinositide 3-kinase ( PI3K ) hyperactivation are common in GBM , promoting tumor growth and survival , including through sterol regulatory element-binding protein 1 ( P36956 ) -dependent lipogenesis . The role of cholesterol metabolism in GBM pathogenesis , its association with P00533 /PI3K signaling , and its potential therapeutic targetability are unknown . In our investigation , studies of GBM cell lines , xenograft models , and GBM clinical samples , including those from patients treated with the P00533 tyrosine kinase inhibitor lapatinib , uncovered an EGFRvIII-activated , PI3K/ P36956 -dependent tumor survival pathway through the low-density lipoprotein receptor ( P01130 ) . Targeting P01130 with the liver X receptor ( LXR ) agonist GW3965 caused inducible degrader of P01130 ( Q8WY64 ) -mediated P01130 degradation and increased expression of the O95477 cholesterol efflux transporter , potently promoting tumor cell death in an in vivo GBM model . These results show that EGFRvIII can promote tumor survival through PI3K/ P36956 -dependent upregulation of P01130 and suggest a role for LXR agonists in the treatment of GBM patients . Human cardiovascular disease IBC chip-wide association with weight loss and weight regain in the look AHEAD trial . BACKGROUND/AIMS : The present study identified genetic predictors of weight change during behavioral weight loss treatment . METHODS : Participants were 3,899 overweight/obese individuals with type 2 diabetes from Look AHEAD , a randomized controlled trial to determine the effects of intensive lifestyle intervention ( ILI ) , including weight loss and physical activity , relative to diabetes support and education , on cardiovascular outcomes . Analyses focused on associations of single nucleotide polymorphisms ( SNPs ) on the Illumina CARe iSelect ( IBC ) chip ( minor allele frequency > 5 % ; n = 31,959 ) with weight change at year 1 and year 4 , and weight regain at year 4 , among individuals who lost ≥ 3 % at year 1 . RESULTS : Two novel regions of significant chip-wide association with year-1 weight loss in ILI were identified ( p < 2.96E-06 ) . O95342 rs484066 was associated with 1.16 kg higher weight per minor allele at year 1 , whereas Q9Y6Q6 , or Q9Y6Q6 , rs17069904 was associated with 1.70 kg lower weight per allele at year 1 . CONCLUSIONS : This study , the largest to date on genetic predictors of weight loss and regain , indicates that SNPs within O95342 , related to bile salt transfer , and Q9Y6Q6 , implicated in adipose tissue physiology , predict the magnitude of weight loss during behavioral intervention . These results provide new insights into potential biological mechanisms and may ultimately inform weight loss treatment . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . β-Arrestin-1 mediates thyrotropin-enhanced osteoblast differentiation . DB00024 ( DB00024 ) activation of the DB00024 receptor ( P16473 ) , a 7-transmembrane-spanning receptor ( 7TMR ) , may have osteoprotective properties by direct effects on bone . P16473 activation by DB00024 phosphorylates protein kinases P31749 , p38α , and P27361 /2 in some cells . We found DB00024 -induced phosphorylation of these kinases in 2 cell lines engineered to express TSHRs , human embryonic kidney P29320 - P16473 cells and human osteoblastic U2OS- P16473 cells . In U2OS- P16473 cells , DB00024 up-regulated pAKT1 ( 7.1±0.5-fold ) , p38α ( 2.9±0.4-fold ) , and pERK1/2 ( 3.1±0.2-fold ) , whereas small molecule P16473 agonist P06681 had no or little effect on pAKT1 ( 1.8±0.08-fold ) , p38α ( 1.2±0.09-fold ) , and pERK1/2 ( 1.6±0.19-fold ) . Furthermore , DB00024 increased expression of osteoblast marker genes P05186 ( 8.2±4.6-fold ) , O14788 ( 21±5.9-fold ) , and osteopontin ( P10451 ; 17±5.3-fold ) , whereas P06681 had little effect ( P05186 , 1.7±0.5-fold ; O14788 , 1.3±0.6-fold ; and P10451 , 2.2±0.7-fold ) . β-Arrestin-1 and -2 can mediate activatory signals by 7TMRs . DB00024 stimulated translocation of β-arrestin-1 and -2 to P16473 , whereas P06681 failed to translocate either β-arrestin . Down-regulation of β-arrestin-1 by siRNA inhibited DB00024 -stimulated phosphorylation of P27361 /2 , p38α , and P31749 , whereas down-regulation of β-arrestin-2 increased phosphorylation of P31749 in both cell types and of P27361 /2 in P29320 - P16473 cells . Knockdown of β-arrestin-1 inhibited DB00024 -stimulated up-regulation of mRNAs for P10451 by 87 ± 1.7 % and O14788 by 73 ± 2.4 % , and P10451 secretion by 74 ± 10 % . We conclude that DB00024 enhances osteoblast differentiation in U2OS cells that is , in part , caused by activatory signals mediated by β-arrestin-1 . 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 . [ Screening of proteins regulated by Q9UL16 gene in nasopharyngeal carcinoma using proteomics technology ] . OBJECTIVE : To screen the proteins under regulation by the candidate tumor suppressor gene Q9UL16 using proteomics technology in nasopharyngeal carcinoma ( NPC ) . METHODS : The cellular proteins were extracted from 3D8 NPC cells with Q9UL16 overexpression and the control P13671 NPC cells . Two-dimensional ( 2D ) gel electrophoresis was employed to compare the protein expression profiles between these two cells , and the differential proteins were identified using peptide mass fingerprinting and database searching . Real-time PCR and Western blotting were used to validate the expression levels of the differential proteins . RESULTS : Matrix-assisted laser desorption/time of flight showed that 3 differential proteins , namely P49327 , P07339 and P00558 , were down-regulated by -3.28 , -1.64 , and -6.97 folds , respectively , which were confirmed by real-time PCR and Western blotting . CONCLUSION : P49327 , P07339 and P00558 are probably the target proteins regulated by Q9UL16 in NPC . 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 . Role of Mucin 1 and P09466 A in recurrent implantation failure . OBJECTIVE : To evaluate and compare the levels of Mucin 1 ( P15941 ) and P09466 A ( GdA ) in precisely timed endometrial biopsies and blood samples taken from women with recurrent implantation failure , and women with proven fertility , in a control group . DESIGN : Molecular studies in human blood and tissue . SETTING : University hospital . PATIENT(S) : Women with recurrent implantation failure and women with proven fertility . INTERVENTION(S) : Primary endometrial cells and blood samples during the implantation " window " ( between day 7 and day 9 after the surge in luteinizing hormone ) . MAIN OUTCOME MEASURE(S) : Expression of P15941 and GdA in the human endometrium and in blood during the implantation window were analyzed by enzyme-linked immunosorbent assay . Additionally , P15941 and GdA levels in tissue were analyzed by western blot during the same period . RESULT(S) : Both blood and tissue measurements of P15941 and GdA were significantly lower in women with recurrent implantation failure than in fertile women during the implantation window . In addition , we found a highly significant correlation between blood vs. tissue measurements of both P15941 and GdA . CONCLUSION(S) : The present study reveals that blood and tissue levels of P15941 and GdA are much lower in women with Q9HBH0 , compared with those in fertile women . Receptivity can be evaluated with noninvasive blood sampling , rather than more-invasive endometrium sampling , as the blood and tissue measurements of P15941 and GdA are correlated . Q15149 transcript diversity : identification and tissue distribution of variants with distinct first coding exons and rodless isoforms . Q15149 is a widely expressed protein that is very large in size and that has all the attributes of a multifunctional crosslinking and organizing element of the cytoskeleton . It displays a multidomain structure , versatile binding activities , and subcellular localizations that enable it to strengthen cells against mechanical stress forces . Moreover , hereditary gene defects in plectin cause epidermolysis bullosa simplex ( Q9BTE0 ) -MD , a severe skin blistering disease with muscular dystrophy . Here we report the analysis of the exonintron organization of the rat plectin gene and the identification of several different isoforms on the transcriptional level . We show that of 35 coding exons identified , 4 serve as alternative first exons splicing into the same successive exon 2 , which is the first of 7 exons encoding a highly conserved actin-binding domain . RNase protection mapping of transcripts containing 3 of the identified 4 alternate first exons revealed their coexpression in rat glioma P13671 cells and in a series of different rat tissues that we examined . Significant variations in expression levels of first exons indicated the possibility of tissue-specific promoter usage . In addition , plectin splice variants lacking exon 31 ( > 3 kb ) , which encodes the entire rod domain of the molecule , were identified in a variety of rat tissues . This study provides first insights into a complex plectin gene regulatory machinery with similarities to that of dystrophin . 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 . 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 . 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 . Gene expression profiles of DB00171 -binding cassette transporter A and C subfamilies in mouse retinal vascular endothelial cells . The purpose of this study was to quantify gene expression levels of the DB00171 -binding cassette ( DB01048 ) transporter A and C subfamilies O95477 -A9 , and P33527 -6/Mrp1-6 , Q99622 /Mrp7 in mouse retinal vascular endothelial cells ( RVEC ) using a combination of a magnetic isolation method for mouse RVEC and real-time quantitative PCR analysis . The transcript level of endothelial cell markers , such as CD31 , Tie-2 , claudin-5 , occludin , ABCB1a/mdr1a , and Q9UNQ0 , were more than 20-fold higher than those in the non-RVEC fraction , suggesting that RVEC in the RVEC fraction are concentrated at least 20-fold compared with those of the non-RVEC fraction . In the O95477 to A9 families , the transcript level of Q99758 and A9 in the RVEC fraction was 1.2- and 32-fold higher than that in the non-RVEC fraction . Although Q99758 was expressed in both the RVEC and non-RVEC fractions , A9 is predominantly expressed in the RVEC fraction . In the P33527 to P13671 and Q99622 families , the transcript level of O15438 , C4 , and P13671 in the RVEC fraction was 27- , 251- , and 242-fold higher , respectively , than that in the non-RVEC fraction , suggesting that O15438 , C4 , and P13671 are predominantly expressed in the RVEC . In conclusion , Q99758 , Q8IUA7 , O15438 , O15439 , and O95255 mRNAs are predominantly expressed at the inner blood-retina barrier ( inner BRB ) and appear to play a major role in the efflux transport of their substrates at the inner BRB . Impaired adipogenesis and lipolysis in the mouse upon selective ablation of the retinoid X receptor alpha mediated by a tamoxifen-inducible chimeric Cre recombinase ( Cre- P27361 ) in adipocytes . P19793 ( RXRalpha ) is involved in multiple signaling pathways , as a heterodimeric partner of several nuclear receptors . To investigate its function in energy homeostasis , we have selectively ablated the RXRalpha gene in adipocytes of 4-week-old transgenic mice by using the tamoxifen-inducible Cre- P27361 recombination system . Mice lacking RXRalpha in adipocytes were resistant to dietary and chemically induced obesity and impaired in fasting-induced lipolysis . Our results also indicate that RXRalpha is involved in adipocyte differentiation . Thus , our data demonstrate the feasibility of adipocyte-selective temporally controlled gene engineering and reveal a central role of RXRalpha in adipogenesis , probably as a heterodimeric partner for peroxisome proliferator-activated receptor gamma . 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 . Endothelial expression of human O95477 in mice increases plasma HDL cholesterol and reduces diet-induced atherosclerosis . The role of endothelial O95477 expression in reverse cholesterol transport ( RCT ) was examined in transgenic mice , using the endothelial-specific Tie2 promoter . Human O95477 ( hABCA1 ) was significantly expressed in endothelial cells ( EC ) of most tissues except the liver . Increased expression of O95477 was not observed in resident peritoneal macrophages . P02647 -mediated cholesterol efflux from aortic EC was 2.6-fold higher ( P < 0.0001 ) for cells from transgenic versus control mice . On normal chow diet , Tie2 hABCA1 transgenic mice had a 25 % ( P < 0.0001 ) increase in HDL-cholesterol ( HDL-C ) and more than a 2-fold increase of P29474 mRNA in the aorta ( P < 0.04 ) . After 6 months on a high-fat , high-cholesterol ( HFHC ) diet , transgenic mice compared with controls had a 40 % increase in plasma HDL-C ( P < 0.003 ) and close to 40 % decrease in aortic lesions ( P < 0.02 ) . Aortas from HFHC-fed transgenic mice also showed gene expression changes consistent with decreased inflammation and apoptosis . Beneficial effects of the O95477 transgene on HDL-C levels or on atherosclerosis were absent when the transgene was transferred onto ApoE or Abca1 knockout mice . In summary , expression of hABCA1 in EC appears to play a role in decreasing diet-induced atherosclerosis in mice and is associated with increased plasma HDL-C levels and beneficial gene expression changes in EC . 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 . 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 . Down-regulation of cholesterol 7alpha-hydroxylase ( P22680 ) gene expression by bile acids in primary rat hepatocytes is mediated by the c-Jun N-terminal kinase pathway . DB04540 7alpha-hydroxylase ( P22680 ) , the rate-limiting enzyme in the neutral pathway of bile acid biosynthesis , is feedback-inhibited at the transcriptional level by hydrophobic bile acids . Recent studies show that bile acids are physiological ligands for farnesoid X receptor ( Q96RI1 ) . Activated Q96RI1 indirectly represses P22680 transcription through induction of small heterodimer protein ( Q15466 -1 ) . In this study , we provide evidence that bile acids rapidly down-regulate P22680 transcription via activation of the JNK/c-Jun pathway . Furthermore , we demonstrate that Q15466 -1 is also a direct target of activated c-Jun . In primary rat hepatocyte cultures , taurocholate ( TCA ) strongly activated JNK in a time- and concentration-dependent manner . P01375 -alpha , a potent activator of JNK , also rapidly activated JNK and down-regulated P22680 mRNA levels . Overexpression of dominant-negative P45983 or a transactivating domain mutant of c-Jun significantly blocked the ability of TCA to down-regulate P22680 mRNA . In contrast , overexpression of wild-type c-Jun ( c-Jun(wt) ) enhanced the repression of P22680 by TCA . Moreover , overexpression of c-Jun(wt) resulted in increased Q15466 -1 promoter activity . Mutation of a putative AP-1 ( c-Jun ) element suppressed c-Jun-mediated activation of the Q15466 -1 promoter construct . These results indicate that the bile acid-activated JNK pathway plays a pivotal role in regulating P22680 levels in primary rat hepatocytes . 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 % . 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 . DB04858 plus cisplatin and irradiation in a mouse model : improved tumor control at the cost of increased toxicity . PURPOSE : DB04858 ( TPZ ) reportedly enhances the tumor cell killing effect of cisplatin up to fivefold and it is an attractive drug for combination with radiotherapy . We evaluated the toxicity of a fractionated combined treatment . METHODS : Murine Q9HBH0 -1 fibrosarcomas growing on the right hind foot of P01024 -H mice were used . Within 2 weeks , animals were treated with six i.p. injections of TPZ ( 43.2-172.8 mg/kg total ) , and/or cisplatin ( 24 mg/kg total ) and ten fractions of 2 Gy to the tumor . All treatments were carried out under anesthesia . Maximum follow-up was 35 days . The local tumor control was determined by calculating the tumor doubling time t ( 2vo ) . In addition to standard toxicity assessment , the major inner organs were examined histologically . RESULTS : The administration of low TPZ doses to the cisplatin/radiotherapy treatment caused only little changes in tumor doubling time ( t ( 2vo ) ) and led to a lethality rate of 15-30 % . Higher TPZ doses caused an increase in t ( 2vo ) , but also a further increase in lethality and toxicity in particular to the heart , liver , kidney and stomach . DB00515 /radiotherapy treatment without TPZ produced no severe toxicity . CONCLUSIONS : This is a detailed study of both the acute and delayed toxicities of combined TPZ treatment in a mouse model . In our study the addition of TPZ to the cisplatin/radiotherapy treatment caused a significant increase in toxicity with only moderate effect on the tumor . 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 . 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 . 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 . 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 . Puerarin decreases serum total cholesterol and enhances thoracic aorta endothelial nitric oxide synthase expression in diet-induced hypercholesterolemic rats . Hypercholesterolemia is a dominant risk factor for the development and progression of atherosclerosis and cardiovascular diseases . Natural compounds have been proved to be useful in lowering serum cholesterol to slow down the progression of cardiovascular diseases . Pueraria lobata is employed clinically to treat cardiovascular diseases in China . In the present study , the atheroscleroprotective potential of the herb 's major active compound , puerarin , was investigated by monitoring serum lipid profile and major enzyme expressions on cholesterol homeostasis in Sprague-Dawley rats fed with control diet , hypercholesterolmic diet or hypercholesterolmic diet plus administration of puerarin ( 300 mg/kg/day , p.o. ) for 4 weeks . Puerarin markedly attenuated the increased total cholesterol induced by hypercholesterolmic diet in both serum and liver . It caused a significant reduction in the atherogenic index . Expression of mRNA for hepatic 7alpha-hydroxylase ( P22680 ) was significantly enhanced but not for those of 3-hydroxy-3-methylglutaryl- DB01992 reductase ( HMGR ) and lanosterol 14alpha-demethylase ( Q16850 ) . To further explore the atheroscleroprotective potential of puerarin , acetylcholine induced endothelium-dependent vasorelaxation and endothelial nitric oxide synthase ( P29474 ) expression on isolated thoracic aortas were analyzed . Animals administered with puerarin suppressed the hypercholesterolemic diet induced impairment of P29474 expression , whereas there was no significant difference in the endothelium-dependent vasorelaxation among various groups of animals . These data indicated that puerarin reduced the atherogenic properties of dietary cholesterol in rats . Its hypocholesterolemic function may be due to the promotion of cholesterol and bile acids excretion in liver . Whether puerarin targets directly on cholesterol homeostasis or both cholesterol homeostasis and endothelial function remains to be determined . Prevalence of genetic risk factors for coronary artery disease in Corsica island ( France ) . We have investigated the frequencies of seven markers among 100 unrelated individuals with angiographically documented CAD ( Coronary Artery Disease ) and among 100 unrelated healthy blood donors in the central region of Corsica island ( France ) . The seven polymorphisms analyzed were chosen from six candidate genes involved in ( 1 ) P00797 -Angiotensin system : Angiotensin converting enzyme ( P12821 I/D ) , ( 2 ) Lipid metabolism : DB04540 Ester Transfer Protein gene ( P11597 TAQ1B ) , ( 3 ) Platelet aggregation : alpha and beta subunits of the platelet GpIIb/GpIIIa integrin complex ( GpIIb HPA3 and GpIIIa Pl(A1/A2) ) , ( 4 ) Coagulation fibrinolysis : P00747 Activator Tissue ( P00750 TPA25 I/D ) and Methylenetetrahydrofolate Reductase ( P42898 C677T and A1298C ) . The samples were genotyped using the polymerase chain reaction followed by restriction enzyme analysis for the RFLPs . No significant difference in allele frequencies between patient and control groups was observed . The occurrence of the P42898 T677T genotype and of the T677T/A1298A compound genotype is higher in cases ( 20 % ) than in the controls ( 4 % ) . Odds ratio seems to indicate that individuals with the P42898 T677T genotype and the T677T/A1298A compound genotype had a 6-fold increased risk for developing CAD ( ORs = 6 ; 95 % CIs = 1.96-18.28 ) suggesting a possible association of P42898 C677T with the risk of CAD in Corsican population . Preparation of phosphorylated starch by dry-heating in the presence of pyrophosphate and its calcium-phosphate solubilizing ability . Starch was phosphorylated through dry-heating in the presence of pyrophosphate at various conditions , and the characteristics of phosphorylated starch ( PS ) were examined . Starch phosphorylation increases as the pH increases from 3 to 6 , but diminishes at pH 7 . Increased temperatures enhance phosphorylation . Data from (31)P NMR suggests that starch phosphorylation occurs mainly at the P01024 -OH and P13671 -OH of the glucose residue . The phosphate linkage is mainly due to monostarch monophosphate . Although starch had almost no calcium phosphate-solubilising capacity , this capacity was markedly enhanced by phosphorylation . X-ray diffraction analysis indicates that the crystal structure of hydroxyapatite was not present in the calcium phosphate-PS complex . Phytoestrogens and their human metabolites show distinct agonistic and antagonistic properties on estrogen receptor alpha ( ERalpha ) and ERbeta in human cells . Phytoestrogens exert pleiotropic effects on cellular signaling and show some beneficial effects on estrogen-dependent diseases . However , due to activation/inhibition of the estrogen receptors ERalpha or ERbeta , these compounds may induce or inhibit estrogen action and , therefore , have the potential to disrupt estrogen signaling . We performed a comprehensive analysis and potency comparison of phytoestrogens and their human metabolites for ER binding , induction/suppression of ERalpha and ERbeta transactivation , and coactivator recruitment in human cells . The soy-derived genistein , coumestrol , and equol displayed a preference for transactivation of ERbeta compared to ERalpha and were 10- to 100-fold less potent than diethylstilbestrol . In contrast , zearalenone was the most potent phytoestrogen tested and activated preferentially ERalpha . All other phytoestrogens tested , including resveratrol and human metabolites of daidzein and enterolactone , were weak ER agonists . Interestingly , the daidzein metabolites 3',4',7-isoflavone and 4',6,7-isoflavone were superagonists on ERalpha and ERbeta . All phytoestrogens tested showed reduced potencies to activate ERalpha and ERbeta compared to diethylstilbestrol on the estrogen-responsive P01024 promoter compared to a consensus estrogen response element indicating a degree of promoter dependency . Zearalenone and resveratrol were antagonistic on both ERalpha and ERbeta at high doses . The phytoestrogens enhanced preferentially recruitment of GRIP1 to ERalpha similar to 17beta-estradiol . In contrast , for ERbeta no distinct preference for one coactivator ( GRIP1 or Q15788 ) was apparent and the overall coactivator association was less pronounced than for ERalpha . Due to their abundance and (anti)-estrogenic potencies , the soy-derived isoflavones , coumestrol , resveratrol , and zearalenone would appear to have the potential for effectively functioning as endocrine disruptors . Carbon-ion beam irradiation kills X-ray-resistant p53-null cancer cells by inducing mitotic catastrophe . BACKGROUND AND PURPOSE : To understand the mechanisms involved in the strong killing effect of carbon-ion beam irradiation on cancer cells with P04637 tumor suppressor gene deficiencies . MATERIALS AND METHODS : DNA damage responses after carbon-ion beam or X-ray irradiation in isogenic HCT116 colorectal cancer cell lines with and without P04637 ( p53+/+ and p53-/- , respectively ) were analyzed as follows : cell survival by clonogenic assay , cell death modes by morphologic observation of DAPI-stained nuclei , DNA double-strand breaks ( DSBs ) by immunostaining of phosphorylated P16104 ( γ P16104 ) , and cell cycle by flow cytometry and immunostaining of Ser10-phosphorylated histone H3 . RESULTS : The p53-/- cells were more resistant than the p53+/+ cells to X-ray irradiation , while the sensitivities of the p53+/+ and p53-/- cells to carbon-ion beam irradiation were comparable . X-ray and carbon-ion beam irradiations predominantly induced apoptosis of the p53+/+ cells but not the p53-/- cells . In the p53-/- cells , carbon-ion beam irradiation , but not X-ray irradiation , markedly induced mitotic catastrophe that was associated with premature mitotic entry with harboring long-retained DSBs at 24 h post-irradiation . CONCLUSIONS : Efficient induction of mitotic catastrophe in apoptosis-resistant p53-deficient cells implies a strong cancer cell-killing effect of carbon-ion beam irradiation that is independent of the p53 status , suggesting its biological advantage over X-ray treatment . Update on denosumab treatment in postmenopausal women with osteoporosis . DB06643 , a fully human recombinant monoclonal antibody to the receptor activator of nuclear factor-κB ligand ( O14788 ) , blocks binding of O14788 to the Q9Y6Q6 receptor , found on the surface of osteoclasts and osteoclast precursors , resulting in decreased bone resorption . Subcutaneous denosumab administration once every 6 months increases bone mineral density at the lumbar spine , total hip , and/or femoral neck , and reduces markers of bone turnover significantly in postmenopausal women with osteoporosis . Relative to placebo , denosumab treatment reduces the risk of vertebral , nonvertebral , and hip fractures significantly . The benefits of denosumab treatment are generally obvious after the first dose and were continued for up to 8 years of treatment in an extension study . The tolerability profile of denosumab during this extension phase was consistent with that observed during the initial 3-year FREEDOM trial . Postmarketing safety surveillance has not shown any unexpected findings . Ongoing safety surveillance will more fully define the long-term safety of denosumab . The benefits of denosumab would seem to be greater than its risks . DB06643 is an important choice in the treatment of postmenopausal women with osteoporosis at increased risk of fractures , including older patients who have difficulty with oral bisphosphonate intake and patients who are intolerant of , or unresponsive to , other therapies . 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 . 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 . 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 . Both P00533 kinase and Src-related tyrosine kinases regulate human ether-à-go-go-related gene potassium channels . Human ether-à-go-go-related gene ( hERG or Kv11.1 ) encodes the rapidly activated delayed rectifier K(+) current ( I(Kr) ) in the human heart . Potential regulation of hERG channel by protein tyrosine kinases ( PTKs ) is not understood . The present study was designed to investigate whether this channel is modulated by PTKs using whole-cell patch clamp technique , and immunoprecipitation and Western blot analysis in P29320 293 cells stably expressing hERG gene . We found that the broad-spectrum PTK inhibitor genistein ( 30 microM ) , the selective P00533 ( epidermal growth factor receptor ) kinase inhibitor AG556 ( 10 microM ) and the Src-family kinase inhibitor Q99463 ( 10 microM ) remarkably inhibited hERG channel current ( I(hERG) ) , and the effects were significantly countered by the protein tyrosine phosphatase ( PTP ) inhibitor orthovanadate ( 1 mM ) . Immunoprecipitation and Western blot analysis demonstrated that membrane protein tyrosine phosphorylation of hERG channels was reduced by genistein , AG556 , and Q99463 . The reduction of hERG channel phosphorylation level by genistein , AG556 or Q99463 was antagonized by orthovanadate . Single point mutation(s) of Y475A and/or Y611A dramatically attenuated the inhibitory effect of I(hERG) by Q99463 and/or AG556 . Our results demonstrate the novel information that I(hERG) is modulated not only by Src-family kinases , but also by P00533 kinases . Y475 and/or Y611 are likely the preferred phosphorylation sites . Regulation of hERG channels by PTKs modifies the channel activity and thus likely alters electrophysiological properties including action potential duration and cell excitability in human heart and neurons . Dacomitinib ( PF-00299804 ) , an irreversible Pan-HER inhibitor , inhibits proliferation of P04626 -amplified breast cancer cell lines resistant to trastuzumab and lapatinib . The human P01133 ( HER ) family of receptors has been pursued as therapeutic targets in breast cancer and other malignancies . DB00072 and lapatinib are standard treatments for P04626 -amplified breast cancer , but a significant number of patients do not respond or develop resistance to these drugs . Here we evaluate the in vitro activity of dacomitinib ( PF-00299804 ) , an irreversible small molecule pan-HER inhibitor , in a large panel of human breast cancer cell lines with variable expression of the HER family receptors and ligands , and with variable sensitivity to trastuzumab and lapatinib . Forty-seven human breast cancer and immortalized breast epithelial lines representing the known molecular subgroups of breast cancer were treated with dacomitinib to determine IC(50) values . P04626 -amplified lines were far more likely to respond to dacomitinib than nonamplified lines ( RR , 3.39 ; P < 0.0001 ) . Furthermore , P04626 mRNA and protein expression were quantitatively associated with response . Dacomitinib reduced the phosphorylation of P04626 , P00533 , Q15303 , AKT , and P29323 in the majority of sensitive lines . Dacomitinib exerted its antiproliferative effect through a combined G(0)-G(1) arrest and an induction of apoptosis . Dacomitinib inhibited growth in several P04626 -amplified lines with de novo and acquired resistance to trastuzumab . Dacomitinib maintained a high activity in lines with acquired resistance to lapatinib . This study identifies P04626 -amplified breast cancer lines as most sensitive to the antiproliferative effect of dacomitinib and provides a strong rationale for its clinical testing in P04626 -amplified breast cancers resistant to trastuzumab and lapatinib . Growth factor receptor/steroid receptor cross talk in trastuzumab-treated breast cancer . Treatment with tyrosine kinase inhibitors ( TKIs ) including trastuzumab has revolutionized the management of P04626 -positive breast cancer . Recent evaluation of clinical trial data suggests that a subset of P04626 /ER double-positive cancers may not receive significant benefit from the TKI therapy . Here we investigate the cross talk between P04626 and ER in breast cancer and monitor the effect of trastuzumab on the tyrosine kinase effector transcription factor Myc . In P04626 -positive breast cancer patients treated with neoadjuvant trastuzumab , steroid receptor-negative status ( ER and PR negative ) of pre-treatment biopsies predicted pathological complete response ( pCR ) ( n=31 patients , P=0.0486 ) , whereas elevated Myc protein inversely associated with pCR ( P=0.0446 ) . Liquid chromatography mass spectrometry identified the corepressor Q9Y618 as a novel Myc-interacting protein . DB00072 treatment enhanced Myc- Q9Y618 interactions in P04626 -overexpressing breast cancer cells ( LCC1 ) and inhibited expression of the Myc target gene survivin . In P04626 -low , ER-positive steroid-dominant cells ( MCF7 ) , trastuzumab therapy repressed Myc- Q9Y618 interactions and upregulated survivin expression . DB00072 treatment induced ER-CBP interactions , enhanced ER transcriptional activity and upregulated expression of the ER target gene pS2 . The absence of pS2 expression in pre-treatment biopsies predicted pCR to neoadjuvant trastuzumab in breast cancer patients ( n=25 , P=0.0089 ) and pS2 expression associated with residual cancer burden ( P=0.0196 ) . Furthermore , metastatic tissues from patients who had failed trastuzumab therapy were pS2 positive . In P04626 -overexpressing cells , trastuzumab treatment can repress Myc transcriptional activity and clinical response is favorable . However , with co-expression of the steroid pathway , this inhibition is lost and response to treatment is often poor . 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 . 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 . 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 . 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 . 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 . 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 . Peritoneal macrophages mediated delivery of chitosan/siRNA nanoparticle to the lesion site in a murine radiation-induced fibrosis model . BACKGROUND : Radiation-induced fibrosis ( Q9HBH0 ) is a dose-limiting complication of cancer radiotherapy and causes serious problems , i.e. restricted tissue flexibility , pain , ulceration or necrosis . Recently , we have successfully treated Q9HBH0 in a mouse model by intraperitoneal administration of chitosan/siRNA nanoparticles directed towards silencing P01375 alpha in local macrophage populations , but the mechanism for the therapeutic effect at the lesion site remains unclear . METHODS : Using the same murine Q9HBH0 model we utilized an optical imaging technique and fluorescence microscopy to investigate the uptake of chitosan/fluorescently labeled siRNA nanoparticles by peritoneal macrophages and their subsequent migration to the inflamed tissue in the Q9HBH0 model . RESULTS : We observed strong accumulation of the fluorescent signal in the lesion site of the irradiated leg up to 24 hours using the optical imaging system . We further confirm by immunohistochemical staining that Cy3 labeled siRNA resides in macrophages of the irradiated leg . CONCLUSION : We provide a proof-of-concept for host macrophage trafficking towards the inflamed region in a murine Q9HBH0 model , which thereby suggests that the chitosan/siRNA nanoparticle may constitute a general treatment for inflammatory diseases using the natural homing potential of macrophages to inflammatory sites . Synthesis of novel spiropyrazoline oxindoles and evaluation of cytotoxicity in cancer cell lines . A series of novel spiropyrazoline oxindole derivatives was synthesized by 1,3-dipolar cycloaddition reaction . The compounds were screened for their in vitro cytotoxic activity against MCF-7 breast cancer cell line ( estrogen receptor positive ( ER+ ) and human epidermal growth factor receptor 2 negative ( P04626 - ) ) . Of the nineteen spiropyrazoline oxindoles tested , six compounds have a GI50 below 12 μM The most potent compounds in this series were also evaluated against MDA-MB-231 breast cancer cell line ( ER- and P04626 - ) . Two spiropyrazoline oxindoles were highly selective between MCF-7 tumor cells and MDA-MB-231 tumor cells . More importantly , they were noncytotoxic against P29320 293T non tumor derived cell lines . P49327 inhibition induces differential expression of genes involved in apoptosis and cell proliferation in ocular cancer cells . P49327 ( P49327 ) , a lipogenic multienzyme complex , is overexpressed in the ocular cancer , retinoblastoma , and is strongly correlated with tumor invasion . Dietary nutrients are reported to exert anticancer effects through inhibition of lipid metabolism . Differential gene expression in cultured retinoblastoma cells induced by cerulenin , a chemical inhibitor of P49327 , was evaluated by cDNA microarray analysis . Cerulenin treatment resulted in significant upregulation of cytochrome c ( P99999 ) by 1.2-fold , whereas S-phase kinase-associated protein-2 ( Q13309 ) , a negative regulator of cell cycle , and the lipid metabolic genes ( Q07869 , P19793 , and O00763 ) were significantly downregulated by -1.59- , -1.8- , -1.83- , and -1.5-fold , respectively , in comparison with untreated cancer cells . The expressions of key differentially expressed genes were confirmed by quantitative real-time PCR . The altered expression of genes involved in cell proliferation , cell signaling , apoptosis , and cell cycle , correlated with the anticancer effects of cerulenin . P49327 inhibition may thus be a potential strategy in retinoblastoma management . MicroRNAs are associated with human embryo implantation defects . BACKGROUND : Repeated implantation failure ( Q9HBH0 ) is a major problem encountered in IVF . We have previously reported that Q9HBH0 -IVF patients have a different endometrial gene expression profile during the window of implantation . Considering microRNA ( miRNA ) function in post-transcriptional regulation of gene expression , the aim of the study was to evaluate the involvement of miRNA in defects of endometrial receptivity . METHODS : We used TaqMan miRNA array cards to identify the miRNAs differentially expressed in the secretory endometrium of Q9HBH0 -IVF patients when compared with fertile women , and bioinformatics tools to identify their predicted targets and the molecular networks they may affect . RESULTS : Comparing miRNA expression profiles , we identified 13 miRNAs , differentially expressed in Q9HBH0 endometrial samples , that putatively regulate the expression of 3800 genes . We found that 10 miRNAs were overexpressed ( including miR 145 , 23b and 99a ) and 3 were underexpressed . Using our previous gene expression analysis , we paralleled miRNA-mRNA expression profiling . By this means , we identified novel and previously characterized miRNA-regulated molecular pathways such as adherens junctions , cell adhesion molecules , Wnt-signaling , p53 signaling and cell cycle pathways . Consistent with the miRNA-predicted targets , mRNA levels of P19022 , P16104 , netrin-4 and secreted frizzled-related protein-4 , belonging to the cell adhesion molecules , Wnt signaling and cell cycle pathways were lower in Q9HBH0 -IVF patients . CONCLUSIONS : To our knowledge , this is the first study to evaluate the differential expression of miRNAs in the secretory endometrium of Q9HBH0 -IVF patients . We suggest that the Q9HBH0 -associated miRNAs could be exploited as new candidates for diagnosis and treatment of embryo implantation failures . 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 . Translatability scoring in drug development : eight case studies . Translational medicine describes the transfer of basic in vitro and in vivo data into human applications . In the light of low rates of market approvals for new medical entities , better strategies to predict the risk of drug development should be used to increase output and reduce costs . Recently , a scoring system to assess the translatability of early drug projects has been proposed . Here eight drugs from different therapeutic areas have been subjected to a retrospective test-run in this system fictively located at the phase II-III transition . The scores gained here underline the importance of biomarker quality which is pivotal to decrease the risk of the project in all cases . This is particularly evident for gefitinib . The P00533 mutation status is a breakthrough biomarker to predict therapeutic success which made this compound clinically acceptable , and this is plausibly reflected by a considerable increase of the translatability score . For psychiatric and Alzheimer 's drugs , and for a P11597 -inhibitor , the lack of suitable biomarkers and animal models is reflected by a low translatability score , well correlating with the excessive translational risk in these areas . These case studies document the apparent utility of the scoring system , at least under retrospective conditions , as the scores correlate with the outcomes at the level of market approval . Prospective validation is still missing , but these case studies are encouraging . 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 . 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 .

MH_train_1153

MH_train_1153

MH_train_1153

interacts_with DB08895?

multiple_choice

Comparison of the micro- and macro-vascular effects of glimepiride and gliclazide in metformin-treated patients with Type 2 diabetes : a double-blind , crossover study . AIMS : To compare the metabolic and vascular effects of two sulphonylureas ( SU ) , gliclazide ( specific for the pancreatic [ Q09428 ] receptor ) and glimepiride ( a nonspecific agent that also binds to vascular and cardiac [ SUR2 ] receptors ) , during chronic administration in metformin-treated patients with Type 2 diabetes ( T2DM ) . METHODS : A randomized , double-blind , crossover study of gliclazide 80 mg P55957 and glimepiride 2 mg OD , each for 4 weeks as add-on therapy to metformin , with a 4-week washout period . Patients attended four study mornings after first dose and 4 weeks ' SU treatment for measurements of arterial distensibility ( Ax ) , pressor responsiveness to i.v. angiotensin II ( ANGII ) , and cutaneous microvascular vasodilator responses to iontophoresis of acetylcholine ( ACh ) and sodium nitroprusside ( SNP ) . RESULTS : Glycaemic responses were similar ( e.g. serum fructosamine was 315 vs 329 micro mol l-1 after 4 weeks ) , and there was no change in augmentation index during treatment with either SU ( 9.1 vs 9.8 mmHg after 4 weeks [ 95 % confidence interval -8.1 , 10.5 ] ) . Similarly , there were no differences between treatments in pressor responsiveness ( e.g. PD10 [ dose of agonist required to increase mean BP by 10 mmHg ] for ANGII was 1.37 vs 1.68 ng kg-1 min-1 [ -4.3 , 6.9 ] ) or cutaneous microvascular vasodilator responses ( peak ACh response 68 +/- 36 vs 63 +/- 34 perfusion units [ -82.7 , 79.1 ] ) . CONCLUSIONS : There is no evidence that Q09428 -specific and nonspecific SUs have differential effects on arterial distensibility , endothelial function or vasodilator mechanisms in metformin-treated patients with T2DM . 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 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 . DB00877 effects on P42345 signaling in benign , premalignant and malignant human breast epithelial cells . DB00877 , an inhibitor of P42345 , is in clinical trials for treatment of cancer . DB00877 resistance has been reported in human breast epithelial tumor cells . DB00877 effects on P42345 signaling and resistance were examined using benign , premalignant and tumor human breast epithelial cells . DB00877 inhibition of cell proliferation , the cell cycle and P42345 signaling , including p70S6 and S6RP phosphorylation , was most effective in benign ( MCF10A ) and premalignant ( MCF10AT ; MCF10ATG3B ) human breast epithelial cells , relative to MCF10CA1a tumor cells . DB00877 resistance was reflected by reduced inhibition of p70S6K and S6RP phosphorylation in MCF10CA1a tumor cells , with RS6P showing the least response to rapamycin in the tumor cells . DB00877 differentially inhibited P40763 phosphorylation in this cell lineage . These data suggest that inhibition of P42345 signaling and P40763 phosphorylation in benign and premalignant cells may be effective in the treatment of proliferative breast disease ( PBD ) and in the prevention of tumorigenesis and tumor recurrence . 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 . Human and rodent pancreatic beta-cells express P05112 receptors and P05112 protects against beta-cell apoptosis by activation of the PI3K and JAK/ P35610 pathways . Secretion of pro-inflammatory cytokines is associated with loss of pancreatic beta-cell viability and cell death . P05112 ( interleukin-4 ) has been reported to mediate a protective effect against the loss of pancreatic beta-cells , and P05112 receptors have been found in rat pancreatic beta-cells at both the RNA and the protein level . The aim of the present study was to investigate P05112 receptor expression in human islet cells and to examine the signalling pathways by which P05112 exerts its effects using the rat beta-cell lines , BRIN-BD11 and P01308 -1E . By means of immunohistochemistry , it was demonstrated that P05112 receptors are present on human islet cells . Using a flow cytometric method for evaluating cell death , it was confirmed that incubating beta-cells with P05112 attenuated cell death induced by IL-1beta and interferon-gamma by approx . 65 % . This effect was abrogated by the presence of the PI3K ( phosphoinositide 3-kinase ) inhibitor , wortmannin , suggesting that activation of the PI3K pathway is involved . In support of this , Western blotting revealed that incubation of cells with P05112 resulted in increased phosphorylation of Akt ( also called protein kinase B ) , a downstream target of PI3K . Increased tyrosine phosphorylation of P42226 ( signal transducer and activator of transcription 6 ) also occurred in response to P05112 and a selective P52333 ( P52333 ) inhibitor reduced the cytoprotective response . Both effects were prevented by overexpression of the tyrosine phosphatase , PTP-BL ( protein tyrosine phosphatase-BL ) . We conclude that P05112 receptors are functionally competent in pancreatic beta-cells and that they signal via PI3K and JAK/ P35610 pathways . These findings may have implications for future therapeutic strategies for the management of diabetes . Generation and properties of a new human ventral mesencephalic neural stem cell line . Neural stem cells ( NSCs ) are powerful research tools for the design and discovery of new approaches to cell therapy in neurodegenerative diseases like Parkinson 's disease . Several epigenetic and genetic strategies have been tested for long-term maintenance and expansion of these cells in vitro . Here we report the generation of a new stable cell line of human neural stem cells derived from ventral mesencephalon ( hVM1 ) based on v-myc immortalization . The cells expressed neural stem cell and radial glia markers like nestin , vimentin and 3CB2 under proliferation conditions . After withdrawal of growth factors , proliferation and expression of v-myc were dramatically reduced and the cells differentiated into astrocytes , oligodendrocytes and neurons . hVM1 cells yield a large number of dopaminergic neurons ( about 12 % of total cells are TH+ ) after differentiation , which also produce dopamine . In addition to proneural genes ( Q9H2A3 , MASH1 ) , differentiated cells show expression of several genuine mesencephalic dopaminergic markers such as : Q8TE12 , O60663 , P48051 , P00325 , P43354 , O75364 , Q05940 and Q01959 , indicating that they retain their regional identity . Our data indicate that this cell line and its clonal derivatives may constitute good candidates for the study of development and physiology of human dopaminergic neurons in vitro , and to develop tools for Parkinson 's disease cell replacement preclinical research and drug testing . DB00877 specifically interferes with GM- P04141 signaling in human dendritic cells , leading to apoptosis via increased p27KIP1 expression . The longevity of dendritic cells ( DCs ) is a critical regulatory factor influencing the outcome of immune responses . Recently , we demonstrated that the immunosuppressive drug rapamycin ( Rapa ) specifically induces apoptosis in DCs but not in other myeloid cell types . The present study unraveled the mechanism used by Rapa to induce apoptosis in human monocyte-derived DCs . Our data demonstrate that granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) preserves DC survival specifically via the phosphatidylinositol-3 lipid kinase/mammalian target of rapamycin ( PI3K/ P42345 ) signaling pathway , which is abrogated by Rapa at the level of P42345 . Disruption of this GM- P04141 signaling pathway induced loss of mitochondrial membrane potential , phosphatidyl-serine exposure , and nuclear changes . Apoptosis of these nonproliferating DCs was preceded by an up-regulation of the cell cycle inhibitor p27( P46527 ) . Overexpression of p27( P46527 ) in DCs using adenoviral gene transduction revealed that apoptosis is directly regulated by p27( P46527 ) . Furthermore , both overexpression of p27( P46527 ) and disruption of the GM- P04141 /PI3K/ P42345 signaling pathway decreased the expression of the antiapoptotic protein mcl-1 . This P42345 /p27( P46527 )/mcl-1 survival seems unique for DCs and may provide novel opportunities to influence immune responses by specific interference with the life span of these cells . 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 . 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 . 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 . 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 . 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 . 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 . Targeting of active P42345 inhibits primary leukemia T cells and synergizes with cytotoxic drugs and signaling inhibitors . OBJECTIVE : Rationally designed therapies aim at the specific disruption of critical signaling pathways activated by malignant transformation or signals from the tumor microenvironment . Because mammalian target of rapamycin ( P42345 ) is an important signal integrator and a key translational regulator , we evaluated its potential involvement in T-cell acute lymphoblastic leukemia ( T-ALL ) and whether P42345 blockade synergizes with chemotherapeutic agents or other signaling antagonists to inhibit primary leukemia T cells . MATERIALS AND METHODS : P42345 signaling status was assessed using biochemical , immunostaining , and molecular regulation studies and functional assays performed to assess the impact of P42345 blockade on T-ALL proliferation , survival , and cell cycle . RESULTS : We observed that P42345 signaling is highly activated in all T-ALL patients tested , with phosphorylation of its downstream substrates eIF4G and S6 ribosomal protein . P42345 activation was detected in vivo and was further increased in vitro by stimulation with interleukin-7 , a potentially leukemogenic cytokine normally produced by the bone marrow microenvironment . In T-ALL cells , P42345 blockade was associated with accumulation of the cyclin-dependent kinase inhibitor p27(kip1) , which preferentially adopted a nuclear localization . Functional studies using rapamycin or CCI-779 showed a dominant inhibitory effect of P42345 blockade on interleukin-7-induced proliferation , survival , and cell-cycle progression of T-ALL cells . Furthermore , P42345 blockade markedly potentiated the antileukemia effects of dexamethasone and doxorubicin , and showed highly synergistic interactions in combination with specific inhibitors of phosphatidylinositol 3-kinase/Akt and P52333 signaling . CONCLUSIONS : This study shows activation of P42345 signaling in primary T-ALL cells evolving in the leukemic bone marrow , and supports the inclusion of P42345 antagonists in current therapeutic regimens for this cancer . P40763 pathway mediates genipin-induced apoptosis in U266 multiple myeloma cells . It has drawn a lot of attention to target signal transducer and activator of transcription 3 ( P40763 ) as a potential strategy for cancer therapeutics . Using several myelogenous cell lines , the effect of genipin ( an active compound of Gardenia fruit ) on the P40763 pathway and apoptosis was investigated . Genipin suppressed the constitutive P40763 activation in U266 and U937 cells and stimulated Src homology 2 domain-containing phosphatase 1 ( Q15466 -1 ) , which dephosphorylates and inactivates P40763 . Specifically , genipin blocked P40763 activation via repressing the activation of c-Src , but not P23458 ( P23458 ) . Genipin also downregulated the expression of P40763 target genes including Bcl-2 , Bcl-x(L) , Survivin , P12004 D1 , and P15692 . Conversely , protein tyrosine phosphatase inhibitor pervanadate blocked genipin induced P40763 inactivation . Using DNA fragmentation or TUNEL assays , we demonstrated the apoptotic effect of genipin on U266 , MM.1S , and U937 cells . Furthermore , genipin effectively potentiated the cytotoxic effect of chemotherapeutic agents , such as bortezomib , thalidomide , and paclitaxel in U266 cells . Our data suggest that through regulation of Src and Q15466 -1 , genipin antagonizes P40763 for the induction of apoptosis in myeloma cells . Enhancement of placenta growth factor expression by oncostatin M in human rheumatoid arthritis synovial fibroblasts . Oncostatin M ( P13725 ) belongs to P05231 subfamily and is mostly produced by T lymphocytes . High levels of P13725 are detected in the pannus of rheumatoid arthritis ( RA ) patients and it may arouse the inflammation responses in joints and eventually leads to bone erosion . P49763 ( P49763 ) is an angiogenic factor and highly homologous with vascular endothelial growth factor ( P15692 ) . It has been recently reported that P49763 is highly expressed in synovial tissue and enhances the production of proinflammatory cytokines including P01375 -α and P05231 . Here , we demonstrated that P13725 increased mRNA and protein levels of P49763 in a time- and concentration-dependent manner in RA synovial fibroblasts . Inhibitors of P52333 and PI3K antagonized P13725 -induced production of P49763 . P13725 enhanced the phosphorylation of Tyr705- P40763 , Ser727- P40763 , Ser473-Akt , and increased the nuclear translocation of phosphorylated P40763 time-dependently . Transfection of dominant negative Akt or application of PI3K inhibitorLY294002 significantly inhibited p-Tyr705- P40763 , p-Ser727- P40763 , and P49763 expression , indicating that Akt is involved in P52333 / P40763 / P49763 signaling cascade . To further examine whether P40763 binds to the promoter region of P49763 , Chip assay was used and it was found that P13725 could bind with P49763 promoter , which was inhibited by P52333 and PI3K inhibitors . Accumulation of P49763 in the pannus may contribute to the inflammation , angiogenesis and joints destruction in RA patients . These findings demonstrated the important role of P13725 in the pathology network of RA and provided novel therapeutic drug targets for RA treatment . 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 . 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 . High glucose augments the angiotensin II-induced activation of O60674 in vascular smooth muscle cells via the polyol pathway . Angiotensin II ( Ang II ) , protein kinase C ( PKC ) , reactive oxygen species ( ROS ) generated by NADPH oxidase , the activation of O60674 ( O60674 ) , and the polyol pathway play important parts in the hyperproliferation of vascular smooth muscle cells ( VSMC ) , a characteristic feature of diabetic macroangiopathy . The precise mechanism , however , remains unclear . This study investigated the relation between the polyol pathway , P05771 , ROS , O60674 , and Ang II in the development of diabetic macroangiopathy . VSMC cultured in high glucose ( HG ; 25 mm ) showed significant increases in the tyrosine phosphorylation of O60674 , production of ROS , and proliferation activities when compared with VSMC cultured in normal glucose ( 5.5 mm ( NG ) ) . Both the aldose reductase specific inhibitor ( zopolrestat ) or transfection with aldose reductase antisense oligonucleotide blocked the phosphorylation of O60674 , the production of ROS , and proliferation of VSMC induced by HG , but it had no effect on the Ang II-induced activation of these parameters in both NG and HG . However , transfection with P05771 antisense oligonucleotide , preincubation with a P05771 -specific inhibitor ( LY379196 ) or apocynin ( NADPH oxidase-specific inhibitor ) , or electroporation of NADPH oxidase antibodies blocked the Ang II-induced O60674 phosphorylation , production of ROS , and proliferation of VSMC in both NG and HG . These observations suggest that the polyol pathway hyperactivity induced by HG contributes to the development of diabetic macroangiopathy through a P05771 -ROS activation of O60674 . Chondrocyte autophagy is stimulated by Q9BYW2 dependent AMPK activation and P42345 suppression . The goal of the study is to examine the relationship between the sensor molecules , Hypoxia Inducible Factor-1 ( Q9BYW2 ) , AMP activated Protein Kinase ( AMPK ) and mammalian Target of DB00877 ( P42345 ) in chondrocyte survival and autophagy . We showed that chondrocytes expressed the energy sensor AMPK-1 and that activation increased with maturation . In addition , we showed that thapsigargin treatment activated AMPK and autophagy in a Q9BYW2 -dependent manner . Using serum-starved AMPK-silenced cells , we demonstrated that AMPK was required for the induction of the autophagic response . We also noted a change in chondrocyte sensitivity to apoptogens , due to activation of caspase-8 and cleavage and activation of the pro-apoptotic protein , P55957 . To test the hypothesis that AMPK signaling directly promoted autophagy , we inhibited AMPK activity in P42345 silenced cells and showed that while P42345 suppression induced autophagy , AMPK inhibition did not block this activity . Based on these findings , it is concluded that because of the micro-environmental changes experienced by the chondrocyte , autophagy is activated by AMPK in a Q9BYW2 -dependent manner . 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 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 . 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 . 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 . 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 . 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 . 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 . Genetic factors affecting susceptibility to alcoholic liver disease in an Indian population . INTRODUCTION. Indians are more likely to develop alcoholic cirrhosis compared to Caucasians , though the cause remains obscure . North Indians tend to consume more alcohol than other parts of the country . Genetic factors are likely to play a major role in these observations . This study investigated whether 10 different polymorphisms were associated with alcohol dependence and/or cirrhosis in North Indians . These were in P00325 *2 ( rs1229984 ) , P00326 *2 ( rs698 ) , P05181 *1D , P05181 *5 ( rs3813867 and rs2031920 ) , P01375 -α(rs1800629) , P01375 -α ( rs361525 ) , IL-1β ( rs3087258 ) , CD-14 ( rs2569190 ) , P22301 ( rs1800872 ) and Q9NST1 ( rs738409 ) . MATERIAL AND METHODS . Hundred healthy controls and 120 chronic alcoholics ( 60 alcoholic noncirrhotics and 60 alcoholic cirrhotics ) attending various departments of PGIMER , Chandigarh were genotyped using PCR-RFLP methods . RESULTS . Alcoholic cirrhotics compared to healthy individuals demonstrated a statistically significant increase in Q9NST1 ( 10109G ) allele ( p = 0.037 , OR = 2.12 , 95 % CI 1.29-3.4 ) . Rest of the associations were not significant after correction for multiple testing . CONCLUSION . Q9NST1 10109G predisposed North Indian subjects to alcoholic cirrhosis . The O60674 / P40763 and mitochondrial pathways are essential for quercetin nanoliposome-induced P13671 glioma cell death . The formulation of quercetin nanoliposomes ( QUE-NLs ) has been shown to enhance QUE antitumor activity in P13671 glioma cells . At high concentrations , QUE-NLs induce necrotic cell death . In this study , we probed the molecular mechanisms of QUE-NL-induced P13671 glioma cell death and examined whether QUE-NL-induced programmed cell death involved Bcl-2 family and mitochondrial pathway through P40763 signal transduction pathway . Downregulation of Bcl-2 and the overexpression of Bax by QUE-NL supported the involvement of Bcl-2 family proteins upstream of P13671 glioma cell death . In addition , the activation of O60674 and P40763 were altered following exposure to QUE-NLs in P13671 glioma cells , suggesting that QUE-NLs downregulated Bcl-2 mRNAs expression and enhanced the expression of mitochondrial mRNAs through P40763 -mediated signaling pathways either via direct or indirect mechanisms . There are several components such as ROS , mitochondrial , and Bcl-2 family shared by the necrotic and apoptotic pathways . Our studies indicate that the signaling cross point of the mitochondrial pathway and the O60674 / P40763 signaling pathway in P13671 glioma cell death is modulated by QUE-NLs . In conclusion , regulation of O60674 / P40763 and ROS-mediated mitochondrial pathway agonists alone or in combination with treatment by QUE-NLs could be a more effective method of treating chemical-resistant glioma . DB00877 -Induced apoptosis in P14210 -stimulated lens epithelial cells by AKT/ P42345 , P29323 and O60674 / P40763 pathways . P14210 ( P14210 ) induced the proliferation of lens epithelial cells ( LECs ) and may be a major cause of posterior capsule opacification ( PCO ) , which is the most frequent postoperative complication of cataract surgery . To date , several agents that can block LECs proliferation have been studied , but none have been used in clinic . Recently , accumulating evidence has suggested rapamycin , the inhibitor of P42345 ( mammalian target of DB00877 ) , was associated with the induction of apoptosis in LECs . The purpose of our study was to investigate the potential effects of rapamycin on P14210 -induced LECs and the underlying mechanisms by which rapamycin exerted its actions . Using cell proliferation , cell viability and flow cytometric apoptosis assays , we found that rapamycin potently not only suppressed proliferation but also induced the apoptosis of LECs in a dose-dependent manner under P14210 administration . Further investigation of the underlying mechanism using siRNA transfection revealed that rapamycin could promote apoptosis of LECs via inhibiting P14210 -induced phosphorylation of AKT/ P42345 , P29323 and O60674 / P40763 signaling molecules . Moreover , the forced expression of AKT , P29323 and P40763 could induce a significant suppression of apoptosis in these cells after treatment of rapamycin . Together , these findings suggested that rapamycin-induced apoptosis in P14210 -stimulated LECs is accompanied by inhibition of AKT/ P42345 , P29323 and O60674 / P40763 pathways , which supports its use to inhibit PCO in preclinical studies and provides theoretical foundation for future possible practice . 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 . 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 . DB04216 inhibits a large panel of kinases implicated in cancer cell biology . Flavonoids are polyphenolic secondary metabolites from plants that possess a common phenylbenzopyrone structure ( P13671 - P01024 - P13671 ) . Depending upon variations in their heterocyclic C-ring , flavonoids are categorised into one of the following groups : flavones , flavonols , flavanones , flavanols , anthocyanidins , isoflavones or chalcones . Flavonols include , among others , the molecules quercetin , myricetin and kaempferol . The anticancer activity of flavonols was first attributed to their electron-donating ability , which comes from the presence of phenolic hydroxyl groups . However , an emerging view is that flavonoids , including quercetin , may also exert modulatory actions in cells by acting through the protein kinase and lipid kinase signalling pathways . Data from the current study showed that 2 μM quercetin , a low concentration that represents less than 10 % of its IC50 growth-inhibitory concentration as calculated from the average of eight distinct cancer cell lines , decreased the activity of 16 kinases by more than 80 % , including P00519 , Aurora-A , -B , -C , P49759 , P36888 , P52333 , MET , P51957 , Q8TD19 , O75914 , P11309 , P07949 , FGF-R2 , PDGF-Rα and -Rß . Many of these kinases are involved in the control of mitotic processes . Quantitative video microscopy analyses revealed that quercetin displayed strong anti-mitotic activity , leading to cell death . In conclusion , quercetin partly exerts its anticancer activity through the inhibition of the activity of a large set of kinases . DB04216 could be an interesting chemical scaffold from which to generate novel derivatives possessing various types of anti-kinase activities . 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 . Inorganic lead enhances cytokine-induced elevation of matrix metalloproteinase P14780 expression in glial cells . Inorganic lead ( Pb ) is a ubiquitous environmental contaminant that produces a variety of deleterious effects in the central nervous system ( CNS ) . Matrix metalloproteinases ( MMPs ) , specifically P14780 , induced by inflammatory cytokines , are increasingly being implicated in CNS pathology . The present study demonstrates that low concentrations of either pro-inflammatory cytokines ( P01375 and IL-1beta ) or Pb did not influence the P14780 expression in a glial cell line ( P13671 ) when added separately . However , combined administration of Pb and cytokines induced a marked synergized elevation of P14780 expression in spite of a reduction in the number of glial cells . These results demonstrate a possible new mechanism by which Pb may induce neuropathological processes . 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 . 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 . 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 . Characterization of the insulin-signaling pathway in lacrimal and salivary glands of rats . PURPOSE : P01308 has been acknowledged as a mediator of several physiological events in lacrimal and salivary glands . We investigated the presence of insulin receptors and of insulin-induced autophosphorylation of the insulin receptor and activation of elements involved in the early steps of insulin signaling in lacrimal and salivary glands of rats . METHODS : Lacrimal and salivary glands of Wistar rats were removed and processed for immunohistochemistry using anti-insulin receptor and anti- DB01277 receptor antibodies . The activation of insulin receptors following insulin treatment , and the involvement of insulin receptor substrates-1 and -2 , Shc , O60674 and P35610 -1 , were analyzed by immunoprecipitation , followed by SDS-PAGE and immunoblotting of rat lacrimal and salivary glands after exposure to insulin . RESULTS : P01308 and DB01277 receptors were present in rat lacrimal and salivary glands and were located predominantly in the cytoplasm and plasma membrane . Functional studies demonstrated that insulin induced a dose-dependent phosphorylation of the insulin receptor , IGF-1R , insulin receptor substrates-1 and -2 , Shc , and P35610 -1 . In rats with streptozotocin-induced diabetes mellitus there was a significant reduction in insulin-induced insulin receptor and P35610 -1 phosphorylation in the lacrimal gland but not in the salivary gland ; there was no influence on Shc phosphorylation in either tissue . CONCLUSIONS : The present results indicate that insulin and DB01277 receptors are expressed in lacrimal and salivary glands , and that insulin can induce the phosphorylation of its receptor and activate elements involved in the early steps of insulin signaling in both tissues . 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 . 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 . 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 . Identification of ciliary neurotrophic factor receptor alpha as a mediator of neurotoxicity induced by alpha-synuclein . Accumulating evidence suggests that extracellular alpha-synuclein ( eSNCA ) plays an important role in the pathogenesis of Parkinson 's disease or related synucleinopathies by inducing neurotoxicity directly or indirectly via microglial or astroglial activation . However , the mechanisms by which this occurs remain to be characterized . To explore these mechanisms , we combined three biochemical techniques - stable isotope labeling of amino acid in cell cultures ( SILAC ) , biotin labeling of plasma membrane proteins followed by affinity purification , and analysis of unique proteins binding to P37840 peptides on membrane arrays . The SILAC proteomic analysis identified 457 proteins , of which , 245 or 172 proteins belonged to membrane or membrane associated proteins , depending on the various bioinformatics tools used for interpretation . In dopamine neuronal cells treated with eSNCA , the levels of 86 membrane proteins were increased and 35 were decreased compared with untreated cells . In peptide array analysis , 127 proteins were identified as possibly interacting with eSNCA . Of those , seven proteins were overlapped with the membrane proteins that displayed alterations in relative abundance after eSNCA treatment . One was ciliary neurotrophic factor receptor , which appeared to modulate eSNCA-mediated neurotoxicity via mechanisms related to P23458 / P40763 signaling but independent of eSNCA endocytosis . Tanshinone IIA inhibits constitutive P40763 activation , suppresses proliferation , and induces apoptosis in rat P13671 glioma cells . P40763 ( P40763 ) is usually constitutively activated in a variety of malignancies . Thus , P40763 may be a promising target for treatment of tumor cells . Recently , Tanshinone IIA ( Tan IIA ) , a major active constituent from the root of Salvia miltiorrhiza Bunge , was reported to have apoptosis inducing effects on a large variety of cancer cells . In this study , we evaluate the anti-proliferation and apoptosis inducing effects of Tan IIA on P13671 glioma cells . Cell growth and proliferation were measured by MTT assay , cell apoptosis was observed by flow cytometry and DNA-fragmentation analysis . Further more , we investigated inhibitory effects of Tan IIA on P40763 activity and its downstream targets : Bcl-XL , cyclin D1 . Alteration of P40763 activity was examined by measuring their DNA binding activity and tyrosine phosphorylation . Changes in the expression levels of Bcl-XL and cyclin D1 were examined by Western blot analysis . We found that the cellular growth were inhibited and cell apoptosis were observed after the treatment with Tan IIA . The P40763 activity was significantly reduced by Tan IIA parallel with a significant attenuation of expression of Bcl-XL and cyclin D1 . These results suggest that Tan IIA may serve as an effective adjunctive reagent in the treatment of glioma for its targeting of constitutive P40763 signaling . Targeting P23458 /2 and P42345 in murine xenograft models of Ph-like acute lymphoblastic leukemia . Q9HC73 rearrangements , P23458 /2 point mutations , and O60674 fusion genes have been identified in Philadelphia chromosome ( Ph ) -like acute lymphoblastic leukemia ( ALL ) , a recently described subtype of pediatric high-risk B-precursor ALL ( B-ALL ) which exhibits a gene expression profile similar to Ph-positive ALL and has a poor prognosis . Hyperactive JAK/ P35610 and PI3K/mammalian target of rapamycin ( P42345 ) signaling is common in this high-risk subset . We , therefore , investigated the efficacy of the JAK inhibitor ruxolitinib and the P42345 inhibitor rapamycin in xenograft models of 8 pediatric B-ALL cases with and without Q9HC73 and JAK genomic lesions . DB08877 treatment yielded significantly lower peripheral blast counts compared with vehicle ( P < .05 ) in 6 of 8 human leukemia xenografts and lower splenic blast counts ( P < .05 ) in 8 of 8 samples . Enhanced responses to ruxolitinib were observed in samples harboring JAK-activating lesions and higher levels of P42229 phosphorylation . DB00877 controlled leukemia burden in all 8 B-ALL samples . Survival analysis of 2 representative B-ALL xenografts demonstrated prolonged survival with rapamycin treatment compared with vehicle ( P < .01 ) . These data demonstrate preclinical in vivo efficacy of ruxolitinib and rapamycin in this high-risk B-ALL subtype , for which novel treatments are urgently needed , and highlight the therapeutic potential of targeted kinase inhibition in Ph-like ALL . 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 . Multilocus variable-number tandem repeat analysis : a helpful tool for subtyping French Clostridium difficile PCR ribotype 027 isolates . The objective of this study was to evaluate the usefulness of multilocus variable-number tandem repeat analysis ( MLVA ) for typing and subtyping of Clostridium difficile . Sixty-eight strains were studied , including strains from PCR ribotypes 027 , 078/126 , 014/020/077 , 017 and 023 . The stability of variable-number tandem repeat ( VNTR ) loci was tested by comparing the MLVA results of two strains subcultured 11 times . After DNA extraction , seven tandem repeat loci ( A6 , P33681 , P13671 , E7 , P13726 , Q9UBA6 , Q13585 ) from published MLVA schemes were amplified by PCR and sequenced . The distance between two strains was determined by calculating the summed tandem repeat difference . Genomic diversity was evaluated by using the minimum spanning tree ( Bionumerics 5.1 software program ; Applied Maths ) . Among the 68 C. difficile isolates examined , 65 unique MLVA types were identified , suggesting a high discriminatory power . An overall good agreement was observed between MLVA types and PCR ribotypes . The stability of VNTR loci was good . MLVA could separate isolates of the hypervirulent PCR ribotype 027 clone in several clusters ; all 027 strains isolated within a hospital were grouped in a specific cluster or were placed very close to each other . Results of MLVA confirmed that strains from PCR ribotypes 078 and 126 were closely related although some were located in different branches of the tree . Similar results were observed for most strains from PCR ribotypes 014 , 020 and 077 . This highly discriminatory method is time-consuming and expensive , but is a valuable tool for subtyping of C. difficile , especially of 027 strains . 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 .

MH_train_1154

MH_train_1154

MH_train_1154

interacts_with DB00945?

multiple_choice

Hypothesis : decreased use of pediatric aspirin has contributed to the increasing prevalence of childhood asthma . BACKGROUND : The prevalence of asthma , atopic eczema , and allergic rhinitis has increased over the last three decades in Western countries . Speculation on the causes of this trend have focused on changes in environmental factors . We hypothesize that the decreased use of aspirin in favor of acetaminophen , due to the association of aspirin with Reye 's syndrome during febrile respiratory infections , may be contributing to these trends in the United States . DATA SOURCES : A detailed literature search was conducted utilizing Medline . Studies considered relevant and important involving both humans and animals in English language were used . HYPOTHESIS : In the United States , the documented prevalence of childhood asthma has increased since 1970 , but the rate of this increase accelerated upward beginning in the early 1980s when the use of pediatric aspirin decreased . During the resolution of common respiratory viral infections , prostaglandin E2 ( DB00917 ) is produced through the actions of cyclooxygenase-2 ( P35354 ) . DB00945 , but not acetaminophen , inhibits P35354 activity . As DB00917 promotes TH2 and inhibits THI type cytokine generation , we hypothesize that the decreased use of aspirin may be a factor in facilitating allergic sensitization and asthma by augmenting the relative Q8IXH7 /TH2 cytokine imbalance in genetically predisposed children . CONCLUSION : We have presented an hypothesis based upon epidemiologic trends , known biologic effects of cytokines and DB00917 on allergic sensitization , and a potentially relevant pharmacologic effect of aspirin to explain a component of the increasing prevalence of childhood asthma in the United States . We suggest this theory be examined further in animal models as well as in other countries where the prevalence of childhood asthma is increasing . A teenager with severe asthma exacerbation following ibuprofen . DB00945 -sensitive asthma , aspirin-intolerant asthma , aspirin- ( or non-steroidal anti-inflammatory drug [ NSAID ] ) exacerbated respiratory disease are terms for a disorder commonly described as affecting adults aged > 30y . With this perception , ibuprofen was administered for postoperative pain management to a 17-year-old boy with allergic rhinitis and previous severe asthma ( at a time when well controlled ) , who then had a severe asthma exacerbation . Analysis of the literature in response to this case highlights four points : 1 ) NSAID-exacerbated asthma is not only a disorder of adults ; it occurs in up to of 2 % in asthmatic children , approaching probably 30 % in older children with severe asthma and nasal disease . 2 ) The asthmatic reaction is dose-dependent and can occur with sub-therapeutic doses . Oral NSAID/aspirin challenge should be conducted in an environment where a severe asthma exacerbation can be appropriately managed . 3 ) The therapeutic use of non-selective [ P23219 preferential ] NSAIDs should be avoided when sensitivity is known or suspected in adults and teenagers with severe asthma and chronic rhinosinusitis or nasal polyps . Use of these agents in younger children with mild episodic wheeze is probably safe . 4 ) DB00316 use is probably safe , but aspirin-exacerbated respiratory disease may occur with clinical doses in a subgroup of aspirin-exacerbated respiratory disease patients . P35354 selective inhibitors are probably safe , although this is controversial . Opioids and tramadol are suitable analgesic alternatives for patients with known or suspected susceptibility . DB00644 1 directly affects corpora lutea lifespan in Mediterranean buffalo ( Bubalus bubalis ) during diestrus : presence and in vitro effects on enzymatic and hormonal activities . The expression of gonadotropin-releasing hormone ( P01148 ) receptor ( P30968 ) and the direct role of P01148 on corpora lutea function were studied in Mediterranean buffalo during diestrus . Immunohistochemistry evidenced at early , mid , and late luteal stages the presence of P30968 only in large luteal cells and P01148 in both small and large luteal cells . Real-time PCR revealed P30968 and P01148 mRNA at the three luteal stages , with lowest values in late corpora lutea . In vitro corpora lutea progesterone production was greater in mid stages and lesser in late luteal phases , whereas prostaglandin F2 alpha ( PGF2alpha ) increased from early to late stages , and DB00917 was greater in the earlier-luteal phase . Cyclooxygenase 1 ( prostaglandin-endoperoxide synthase 1 ; P23219 ) activity did not change during diestrus , whereas P35354 increased from early to late stages , and DB00917 -9-ketoreductase ( DB00917 -9-K ) was greater in late corpora lutea . P23219 activity was greater than P35354 in early corpora lutea and lesser in late luteal phase . In corpora lutea cultured in vitro , the P01148 analog ( buserelin ) reduced progesterone secretion and increased PGF2alpha secretion as well as P35354 and DB00917 -9-K activities at mid and late stages . DB00917 release and P23219 activity were increased by buserelin only in late corpora lutea . These results suggest that P01148 is expressed in all luteal cells of buffalo , whereas P30968 is only expressed in large luteal phase . Additionally , P01148 directly down-regulates corpora lutea progesterone release , with the concomitant increases of PGF2alpha production and P35354 and DB00917 -9-K enzymatic activities . [ 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 . Sites of action for future therapy : an adenosine-dependent mechanism by which aspirin retains its antiinflammatory activity in cyclooxygenase-2 and NFkappaB knockout mice . The antiinflammatory action of aspirin is generally attributed to inhibition of cyclooxygenases 1 and 2 , but additional mechanisms are at work . These include inhibition of NFkappaB translocation to the nucleus and the capacity of aspirin to promote accumulation of adenosine , a potent antiinflammatory autocoid . We tested these hypotheses in the murine air pouch model of acute inflammation in wild type mice and in cyclooxygenase 2 or NFkappaB knockouts . The antiinflammatory effects of aspirin , sodium salicylate and indomethacin did not correlate with inhibition of cyclooxygenase in either group . Indeed , aspirin retained its antiinflammatory properties even in P35354 knockouts . Similarly , aspirin was no less antiinflammatory in mice rendered deficient for NFkappaB ( Q14511 ) than in wild type controls . In contrast , dexamethasone lost its antiinflammatory capacity in NFkappaB knockouts . DB00945 and sodium salicylate dramatically increased concentrations of adenosine in exudates , a property shared with methotrexate and sulfasalazine . Removal of adenosine by adenosine deaminase or specific antagonism of adenosine at A(2)receptors completely reversed the antiinflammatory effects of aspirin and sodium salicylate , but not those of dexamethasone . This adenosine-dependent , antiinflammatory effect of aspirin points to another target of drug development . The Effect of Xuefuzhuyu Oral Liquid on DB00945 Resistance and Its Association with rs5911 , rs5787 , and rs3842788 Gene Polymorphisms . DB00945 should be continued indefinitely in patients after interventional therapy , but 10 % to 40 % of patients experience recurrent vascular events despite adequate aspirin therapy , a condition known as aspirin resistance ( AR ) . Xuefuzhuyu oral liquid , derived from the classic recipe Xuefuzhuyu decoction , has been well documented to inhibit platelet aggregation and to improve hemorheology . The aims of this study were to investigate the effects of Xuefuzhuyu oral liquid on AR in patients with chronic stable angina after percutaneous coronary intervention ( P05154 ) and the possible genetic markers related to the drug response . 43 patients diagnosed as having aspirin resistance or semi-resistance were randomly divided into control and treatment groups after screening 207 stable Q8NE62 patients . Platelet aggregation rate was determined using turbidimetry . Three single nucleotide polymorphisms in P23219 ( rs5787 , rs3842788 ) and GP IIb ( rs5911 ) were genotyped in whole blood samples using ABI Q96FA3 7900 HT Fast Real-Time instrument and ABI Q96FA3 3730 DNA Sequencer . The results showed that Xuefuzhuyu oral liquid could effectively improve blood stasis syndrome and AR by inhibiting ADP-induced platelet aggregation and that patients with the rs5911 genetic variant exhibited better drug response upon treatment with Xuefuzhuyu oral liquid , which suggests Xuefuzhuyu oral liquid as a new possible drug for the prevention of AR . 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 . DB00945 and Other P23219 inhibitors . Currently available antiplatelet drugs interfere with the process of platelet activation and aggregation by selectively blocking key enzymes involved in the synthesis of platelet agonists , or membrane receptors mediating activation signals . Pharmacological interference with critical molecular pathways of platelet activation and aggregation may reduce the risk of atherothrombotic complications through mechanisms that are also responsible for an increased risk of bleeding . Acetylsalicylic acid ( aspirin ) represents a prototypic antiplatelet agent . The aim of this chapter is to integrate our current understanding of the molecular mechanism of action of aspirin with the results of clinical trials and epidemiological studies assessing its efficacy and safety . Moreover , the antiplatelet properties of reversible inhibitors of the same drug target will also be reviewed . The effect of aspirin and nonsteroidal anti-inflammatory drugs on prostaglandins . Cyclic prostanoids play important physiologic roles in inflammation and maintaining normal function of several organ systems . Prostaglandin production requires the conversion of arachidonate to the intermediate prostaglandin H2 , by the 2-step cyclo-oxygenation and peroxidation catalyzed by the enzyme cyclo-oxygenase ( also called prostaglandin H synthase ) . DB00945 and nonsteroidal anti-inflammatory drugs ( NSAIDs ) block the production of cyclic prostanoids by binding in different ways to this enzyme and blocking the active site . This results in decreased inflammation , but it can also produce side effects in the gastrointestinal tract , kidney , and platelets . Recent data demonstrate that there are 2 isoforms of the cyclo-oxygenase enzyme , called P23219 and P35354 . These isoforms are similar in size , substrate specificity , and kinetics , but vary in their expression and distribution . Normal physiologic functions appear to be maintained by P23219 , while P35354 appears to mediate the inflammatory response . Nonsteroidal drugs with selective inhibitory activity on the P35354 isoform should theoretically decrease inflammation while maintaining normal physiologic prostaglandin levels . Current NSAIDs are not selective enough to confirm this , but newer , more selective inhibitors of P35354 may answer this important question . 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 . Hyper-IgM , neutropenia , mild infections and low response to polyclonal stimulation : hyper-IgM syndrome or common variable immunodeficiency ? A young woman presenting respiratory infections , polyarthritis , severe neutropenia , and increased serum IgM was treated with intravenous immunoglobulin ( IVIG ) with good clinical and laboratory outcome followed by a loss of efficacy . The increased serum IgM associated to recurrent infections and autoimmune manifestations suggested the diagnosis of a hyper-IgM syndrome ( HIGMs ) . The frequency of peripheral T cells , the expression of P25942 on the patients ' B cells and P29965 on T cells and the activation-induced cytidine deaminase ( Q9GZX7 ) and uracil-DNA glycosylase ( P13051 ) at mRNA level was comparable to controls . In contrast , the frequency of B cells was one half of the healthy control and all cells showed an atypical phenotype . Although Q9GZX7 and P13051 were normal , class-switch recombination was not very efficient because circulating switched memory were reduced and , once stimulated with CpG , generated less antibody-secreting cells than controls . An increase in serum B Lymphocytes stimulator ( Q9Y275 ) was also found . The patient presented a peculiar clinical and immunological phenotype fitting for many aspects of both HIGM4 and Common Variable Immunodeficiency ( CVID ) . These findings underline the need to better explore the complex link between these two diseases . DB00945 -exacerbated asthma . : This review focuses on aspirin-exacerbated asthma ( AEA ) . The review includes historical perspective of aspirin , prevalence , pathogenesis , clinical features and treatment of AEA . The pathogenesis of AEA involves the cyclooxygenase and lipooxygenase pathway . DB00945 affects both of these pathways by inhibiting the enzyme cycooxygenase-1 ( P23219 ) . Inhibition of P23219 leads to a decrease in prostaglandin E2 ( DB00917 ) . The decrease in DB00917 results in an increase in cysteinyl leukotrienes by the lipooxygenase pathway involving the enzyme 5-lipooxygenase ( P09917 ) . Leukotriene C4 ( LTC4 ) synthase is the enzyme responsible for the production of leukotriene C4 , the chief cysteinyl leukotriene responsible for AEA . There have been familial occurences of AEA . An allele of the Q16873 gene in AEA is known as allele C . Allele C has a higher frequency in AEA . Clinical presentation includes a history of asthma after ingestion of aspirin , nasal congestion , watery rhinorrhea and nasal polyposis . Treatment includes leukotriene receptor antagonists , leukotriene inhibitors , aspirin desinsitaztion and surgery . AEA is the most well-defined phenotype of asthma . Although AEA affects adults and children with physician-diagnosed asthma , in some cases there is no history of asthma and AEA often goes unrecognized and underdiagnosed . DB00945 -exacerbated cutaneous disease . It has been recognized that a high proportion of chronic urticaria patients experience symptom aggravation when exposed to aspirin and NSAIDs . This clinical picture is known as DB00945 -exacerbated cutaneous disease . The pathogenesis of these exacerbations is related to the inhibition of cyclooxygenase-1 leading to a decreased synthesis of DB00917 and an increased cysteinyl leukotriene production in the skin and subcutaneous tissues . Patient management comprises the treatment of the underlying cutaneous disease with nonsedating antihistamines and other medications , avoidance of P23219 inhibitors , and the use of alternative NSAIDs that do not inhibit P23219 for the relief of pain , inflammation and fever . Interactive effect of interleukin-6 and prostaglandin E2 on osteoclastogenesis via the O00300 / O14788 / Q9Y6Q6 system . The O00300 / O14788 / Q9Y6Q6 system regulates osteoclastogenesis . Both cyclooxygenase-2 ( P35354 ) /prostaglandin E2 ( DB00917 ) and interleukin-6 ( P05231 ) are reported to induce osteoclast differentiation . The mechanisms underlying these signaling pathways on the O00300 / O14788 / Q9Y6Q6 system are not fully understood . We herein demonstrate that P35354 and DB00917 stimulated osteoclastogenesis through inhibition of O00300 secretion , stimulation of O14788 production by osteoblasts , and upregulation of Q9Y6Q6 expression in osteoclasts . DB00917 also stimulated P05231 production , and P05231 , in turn , increased DB00917 secretion , P35354 , and EP4/EP2 expression in bone cells . These findings provide evidence of interactive effect of DB00917 and P05231 signaling pathways in osteoclastogenesis via effect on the O00300 / O14788 / Q9Y6Q6 system . Nonsteroidal antiinflammatory drugs and cyclooxygenase inhibition in the gastrointestinal tract : a trip from peptic ulcer to colon cancer . DB00945 was commercialized more than a 100 years ago . Today , this compound is still widely prescribed , and new mechanisms of action and indications are being tested . Inhibition of cyclooxygenase ( P36551 ) -1 and P35354 by aspirin or its related compounds , nonsteroidal antiinflammatory drugs ( NSAIDs ) , has been associated with both adverse and beneficial effects in the gastrointestinal ( GI ) tract . Inhibition of P23219 has been linked to GI adverse effects . Adverse effects of NSAIDs and aspirin in the upper GI tract include esophagitis , peptic ulcer , peptic ulcer complications , and death . Effective preventive therapies are available that have been associated with a progressive decline in the rate of hospitalization due to upper GI complications . NSAIDs and aspirin can also damage the small bowel and the colon . NSAID enteropathy is frequent and in most cases subclinical ( increased mucosal permeability , inflammation , erosion , ulcer ) . However , more serious clinical outcomes such as anemia , bleeding , perforation , obstruction , diverticulitis , and deaths have also been described . Prevention therapy of NSAID damage to the lower GI tract is not well defined . Inhibition of P35354 by NSAIDs , coxibs , or aspirin seems to provide beneficial effects to the GI tract . Observational studies show that these compounds reduce the risk of both upper and lower GI cancers . Randomized controlled trials have shown that aspirin and coxibs reduce the recurrence rate of colonic polyps , and long-term cohort studies have shown that aspirin reduces the risk of colon cancer time and dose dependently . New studies will have to define the appropriate population that may benefit with these therapies . NSAIDs and the gastrointestinal tract . NSAIDs incur significant gastrointestinal ( GI ) side effects . The complication risk increases with history of peptic ulcer or older age . Helicobacter pylori infection and cardioprotective aspirin have independent and additive risks in the presence of NSAID use . NSAID enteropathy is increasingly recognized . Cardiovascular and GI risk stratification and H. pylori infection testing should be done before initiating NSAIDs . An NSAID combined with a proton pump inhibitor ( PPI ) is comparable to cyclooxygenase ( P36551 ) -2 inhibitors for gastroprotection , but for high-risk patients , P35354 plus PPI should be considered . DB00945 and P35354 inhibitors are associated with reduced colon adenoma risk , but higher dose and longer duration of treatment with aspirin appears effective . Hence , patients at high risk of colorectal cancer ( with significant family or personal history of premalignant adenoma ) must be identified , and cardiovascular and GI risk must be assessed before using these agents as chemopreventive drugs . Physiological mediators in nonsteroidal anti-inflammatory drugs ( NSAIDs ) -induced impairment of gastric mucosal defense and adaptation . Focus on nitric oxide and lipoxins . Prostaglandins mediate various physiological aspects of mucosal defense and the suppression of prostaglandin synthesis in the stomach is a critical event in terms of the development of mucosal injury after NSAID administration . However , it has become clear that other mediators besides prostaglandins can similarly act to protect the stomach from injury . For instance , nitric oxide ( NO ) released from vascular epithelium , epithelial cells of gastrointestinal tract and sensory nerves can influence many of the same components of mucosal defense as do prostaglandins . Thus , administration of NO in a form of NO-donors exert protective influence on the stomach from the injury that usually occurs when mucosal prostaglandin levels are suppressed . The new class of NO releasing NSAIDs , including NO-aspirin , represent a very promising approach to reducing the toxicity of anti-inflammatory drugs . Lipoxins are another group of lipid mediators that can protect the stomach . DB00945 -triggered lipoxin synthesis , via P35354 , acts to reduce the severity of damage induced by this drug . Lipoxin analogues may prove to be useful for preventing mucosal injury and for modulating mucosal inflammation . DB00945 -triggered lipoxin also seems to play in important role in gastric adaptation during chronic aspirin administration . Suppression of P35354 activity by selective P35354 inhibitors abolishes the production of this endogenous gastroprotective substance and diminishes the gastric tolerability of NSAIDS and gastric adaptation to these drugs . This review was designed to give an updated overview on the physiological factors and experimental and clinical attempts that were used or may be used in the future as the therapeutic approach to counteract adverse effects in the stomach associated with NSAID ingestion . DB00945 and colorectal cancer : back to the future . Abundant epidemiologic evidence indicates that regular and long-term use of aspirin is associated with a significant reduction in the incidence of colorectal cancer . The long duration of aspirin needed to prevent colorectal cancer is believed to be due to inhibition of precursor lesions known as adenomas , the recurrence of which is inhibited by aspirin in randomized trials . DB00945 intake has also been associated with a statistically significant improvement in patient survival after curative resection of colorectal cancer in large observational studies . In these cohorts , the survival benefit of aspirin was shown to depend upon the level of P35354 expression in the primary colorectal cancer . More recent analysis of patient tumors from these observational cohorts suggests that the benefit of aspirin may be limited to specific molecular subtypes . DB00945 intake following colorectal cancer resection was associated with a significant improvement of survival in patients whose tumors carried mutant , but not wild-type , copies of the phosphoinositide 3-kinase ( PI3KCA ) gene , especially tumors that overexpressed P35354 . A mechanistic explanation is suggested by the finding that inhibition of P36551 -mediated prostaglandin E2 synthesis by aspirin attenuates PI3K signaling activity that is known to regulate cancer cell proliferation and survival . DB00945 has also been shown to reduce the incidence of colorectal cancers bearing wild-type , but not mutant alleles of the P15056 (V600E) oncogene . Although provocative , the potential utility of these molecular markers for predicting aspirin efficacy awaits prospective evaluation in clinical trials . If validated , these findings may support a personalized approach to using aspirin for the therapy of colorectal cancer . Clinical pharmacology of cyclooxygenase inhibition and pharmacodynamic interaction with aspirin by floctafenine in Thai healthy subjects . DB08976 , a hydroxyquinoline derivative with analgesic properties , is widely used in Thailand and many other countries . The objectives of this study were to evaluate in Thai healthy volunteers : i ) the inhibition of whole blood cyclooxygenase( P36551 )-2 and P23219 activity by floctafenine and its metabolite floctafenic acid in vitro and ex vivo after dosing with floctafenine ; ii ) the possible interference of floctafenine administration with aspirin antiplatelet effects . We performed an open-label , cross-over , 3-period study , on 11 healthy Thai volunteers , who received consecutively floctafenine(200mg/TID) , low-dose aspirin(81mg/daily) or their combination for 4 days , separated by washout periods . DB08976 and floctafenic acid resulted potent inhibitors of P23219 and P35354 in vitro ( floctafenic acid was more potent than floctafenine ) showing a slight preference for P23219 . After dosing with floctafenine alone , whole blood P23219 and P35354 activities were inhibited ex vivo in a time-dependent fashion which paralleled floctafenic acid plasma concentrations . DB00945 alone inhibited profoundly and persistently platelet P23219 activity and AA-induced platelet aggregation throughout 24-h dosing interval which was affected by the co-administration of floctafenine . At 24 h after dosing with aspirin and floctafenine , the inhibition of platelet thromboxane(TX)B2 generation and aggregation were significantly ( P less than 0.05 ) lower than that caused by aspirin alone . Therapeutic dosing with floctafenine profoundly inhibited prostanoid biosynthesis through the rapid conversion to floctafenic acid . DB08976 interfered with the antiplatelet effect of aspirin . Our results suggest that floctafenine should be avoided in patients with cardiovascular disease under treatment with low-dose aspirin . 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 . Metformin Improves P01308 Signaling in Obese Rats via Reduced IKKbeta Action in a Fiber-Type Specific Manner . Metformin is a widely used insulin-sensitizing drug , though its mechanisms are not fully understood . Metformin has been shown to activate AMPK in skeletal muscle ; however , its effects on the inhibitor of kappaB kinasebeta ( IKKbeta ) in this same tissue are unknown . The aim of this study was to ( 1 ) determine the ability of metformin to attenuate IKKbeta action , ( 2 ) determine whether changes in AMPK activity are associated with changes in IKKbeta action in skeletal muscle , and ( 3 ) examine whether changes in AMPK and IKKbeta function are consistent with improved insulin signaling . Lean and obese male Zuckers received either vehicle or metformin by oral gavage daily for four weeks ( four groups of eight ) . Proteins were measured in white gastrocnemius ( WG ) , red gastrocnemius ( RG ) , and soleus . AMPK phosphorylation increased ( P < .05 ) in WG in both lean ( 57 % ) and obese ( 106 % ) , and this was supported by an increase in phospho-ACC in WG . Further , metformin increased P25963 levels in both WG ( 150 % ) and RG ( 67 % ) of obese rats , indicative of reduced IKKbeta activity ( P < .05 ) , and was associated with reduced P35568 -pSer(307) ( 30 % ) in the WG of obese rats ( P < .02 ) . From these data we conclude that metformin treatment appears to exert an inhibitory influence on skeletal muscle IKKbeta activity , as evidenced by elevated P25963 levels and reduced P35568 - DB00133 (307) phosphorylation in a fiber-type specific manner . 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 . 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 . DB00945 and non-small cell lung cancer resections : effect on long-term survival . OBJECTIVE : Survival after resections for non-small cell lung cancer remains poor . Recurrent lung cancer remains common . Due to the common risk factor of smoking , cardiovascular deaths occur in the absence of recurrent lung cancer in up to 15 % of patients . DB00945 has been proven to reduce cardiovascular mortality as a secondary prophylactic agent , but not as a primary agent . DB00945 being a P35354 inhibitor has been shown to reduce the chance of metastasis in adenocarcinoma but not squamous carcinoma . We sought to investigate the effect of long-term aspirin therapy on survival post potentially curative surgery . METHODS : We analysed a prospective thoracic surgical database , from time period 2003 to date . Patients who were on aspirin pre-operatively , N=412 were compared to non users , N=1353 . Patient long-term outcome was assessed utilising the national strategic tracking service that operates in the United Kingdom . Cox proportional hazards analysis was used to determine significant factors affecting survival . RESULTS : 100 % survival follow up was achieved . Regular users of aspirin had > 5 % increased survival , which was significant , p=0.05 , despite having a higher cardiovascular risk profile . Mode of death data was not available . CONCLUSIONS : Adjuvant aspirin post resection for potentially curative non-small cell lung cancer significantly increases survival . The mechanism of increased survival needs further investigation and is the basis for the trial : Adjuvant DB00945 for Non-Small cell Lung Cancer -- The Big A Trial. www.TheBigATrial.co.uk . A novel family of hydroxamate-based acylating inhibitors of cyclooxygenase . DB00945 irreversibly inhibits cyclooxygenase ( P36551 ) by acetylating a serine residue in the active site . We synthesized a series of novel acylating agents based on our previously reported acetylating compound , O-acetylsalicylhydroxamic acid . One of these , triacetylsalicylhydroxamic acid ( TriAcSHA ) was more effective than aspirin and O-acetylsalicylhydroxamic acid in inactivating both P23219 and P35354 . Preincubation of P23219 with inhibitor for 5 min yielded IC(50) values of 18 microM for TriAcSHA and 60 microM for acetylsalicylic acid . Inhibition was time-dependent , with complete inhibition within 10 min at a concentration of 50 microM . As with aspirin , mutation of the serine 530 of P23219 to alanine abolished the activity of the TriAcSHA . Mutation of the alanine 119 to a glutamine markedly reduced the sensitivity to TriAcSHA , suggesting that this residue was necessary for the interaction with the enzyme . TriAcSHA was also more effective than aspirin as an inhibitor of platelet aggregation induced by arachidonic acid . The diacetylated phenylhydroxamates N-methyl-O,O-diacetylsalicylhydroxamic acid , N,O-diacetylbenzohydroxamic acid , and 2-methyl-O,N-diacetylbenzohydroxamic acid showed reduced or absent activity against P23219 . In addition , we synthesized a series of triacylsalicylhydroxamic acids with progressively longer acyl groups ( three to six carbons ) . All of the compounds inhibited P23219 and demonstrated progressively greater P23219 selectivity with increasing number of carbons . Hence , salicylhydroxamic acid provides a versatile backbone for the generation of a family of acylating inhibitors of cyclooxygenase . 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 . [ 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 . 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 . DB00945 -mediated P35354 transcript stabilization via sustained p38 activation in human intestinal myofibroblasts . Acetylsalicylic acid ( aspirin ) is a cyclooxygenase ( P36551 ) inhibitor , yet some of its therapeutic effects are thought to derive from mechanisms unrelated to prostaglandin synthesis inhibition . In human intestinal myofibroblasts , aspirin , at therapeutic doses , had the unexpected effect of inducing prolonged P35354 expression . This induction was especially pronounced when cells were treated with interleukin-1alpha ( IL-1 ) plus aspirin for 24 h . Sodium salicylate , a poor P36551 inhibitor , likewise enhanced IL-1-mediated P35354 gene expression whereas DB00244 ( DB00244 ) or indomethacin had no effect . The P35354 transcriptional rate , measured by nuclear runoff analysis and heterogeneous nuclear RNA reverse transcription-polymerase chain reaction , was only modestly elevated by aspirin treatment . In contrast , aspirin treatment dramatically stabilized the P35354 message . The P35354 mRNA half-life in IL-1 treated cells was 1 h and was increased in excess of 5 h in IL-1 + aspirin-treated cells . Phosphorylation of p38 MAPK was enhanced in aspirin-treated cells ( but not in cells treated with DB00244 or indomethacin ) for up to 24 h after treatment . Inhibition of p38 activity negated aspirin-mediated P35354 mRNA stabilization and the resultant increase in P35354 mRNA and protein levels . The modest transcriptional response seen in aspirin treated cells was also abolished by p38 inhibition . We conclude that aspirin enhances P35354 expression via sustained activation of p38 , which results in prolonged stabilization of the P35354 message and a slightly elevated transcription rate . DB00945 also enhanced steady-state mRNA levels of other IL-1 modulated genes ( IL-1beta , P05231 , groalpha , and TNFalpha ) that are likewise regulated at the level of message stability via p38 activation . Angioedema associated with aspirin and rofecoxib . OBJECTIVE : To report the probable association of angioedema with aspirin therapy and the selective cyclooxygenase-2 ( P35354 ) inhibitor rofecoxib . CASE SUMMARY : A 44-year-old white woman , previously tolerant to aspirin and other nonsteroidal antiinflammatory drugs ( NSAIDs ) , developed angioedema of the lips after ingesting two 325-mg aspirin tablets during one day . The reaction occurred 3 hours after taking the second aspirin and resolved within 3 hours . Two weeks later , the patient took a 25-mg rofecoxib tablet for a sore throat , and she developed angioedema 5(1/2) hours later . Although the woman took 50 mg of diphenhydramine , the swelling did not subside . She repeated the diphenhydramine dose in the evening and , by noon the next day , 26(1/2) hours after the angioedema began , it was resolved . The patient 's internist prescribed an epinephrine auto-injector and advised her to consult an allergist . With skin testing and oral rechallenge with aspirin , but not rofecoxib , the allergist determined the cause of the reactions to be aspirin-induced angioedema and selective P35354 inhibitor intolerance . The Naranjo probability scale indicated that aspirin was a highly probable cause and rofecoxib was a probable cause of this patient 's angioedema . DISCUSSION : DB00945 -induced angioedema and NSAID intolerance have been well documented . There are reports of both tolerance and intolerance to selective P35354 inhibitors in patients with documented allergy-like reactions to aspirin and NSAIDs . CONCLUSIONS : Patients with aspirin and NSAID intolerance may develop intolerance to P35354 inhibitors , especially with repeated exposure . 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 . 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 . 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 . 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 . DB00945 -induced peptic ulcer and genetic polymorphisms . There are a few studies of the association between genetic polymorphisms and the risks of acetylsalicylic acid (aspirin)-induced ulcer or its complications . Two single nucleotide polymorphisms ( SNP ) of cyclooxygenase-1 ( P23219 ) , A-842G and C50T , exhibited increased sensitivity to aspirin and had lower prostaglandin synthesis capacity , lacking statistical significance in the association with bleeding peptic ulcer . A recent Japanese study indicated that the number of P23219 -1676T alleles was a significant risk factor for peptic ulcer in users of non-steroidal anti-inflammatory drugs ( NSAIDs ) . There are some genetic polymorphisms for aspirin resistance , such as platelet membrane glycoproteins , thromboxane A2 ( TXA2 ) receptor , platelet activating factor acetylhydrolase and coagulation factor XIII ; however , data on the frequency of gastrointestinal ( GI ) events in these variants are lacking . Carrying the P11712 variants is reported a significantly increased risk of non-aspirin NSAID-related GI bleeding . The polymorphisms of interleukin-1beta ( IL-1beta ) and tumor necrosis factor-alpha ( P01375 ) have been associated with development of peptic ulcer or gastric cancer . In a recent investigation , carriage of the IL-1beta-511 T allele was significantly associated with peptic ulcer among low-dose aspirin users . Hypoacidity in corpus gastritis related to polymorphisms of pro-inflammatory cytokines seems to reduce NSAIDs or aspirin-related injury . Data on which polymorphisms are significant risk factors for GI events in aspirin users are still lacking and further large-scale clinical studies are required . 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 . DB00945 -sensitive respiratory disease . DB00945 -sensitive respiratory disease ( ASRD ) is a condition characterized by persistent and often severe inflammation of the upper and lower respiratory tracts . Patients develop chronic eosinophilic rhinosinusitis , nasal polyposis , and asthma . The ingestion of aspirin and other cyclooxygenase-1 ( P23219 ) inhibitors induces exacerbations of airway disease that may be life-threatening . Thus , aspirin sensitivity is a phenotypic marker for the syndrome , yet nearly all affected individuals can be desensitized by the administration of graded doses of aspirin , leading to long-term clinical benefits . Patients with aspirin sensitivity are often able to tolerate selective P35354 inhibitors . The pathogenesis of ASRD is underpinned by abnormalities in eicosanoid biosynthesis and eicosanoid receptor expression coupled with intense mast cell and eosinophilic infiltration of the entire respiratory tract . This review focuses on the molecular , cellular , and biochemical abnormalities characterizing ASRD and highlights unanswered questions in the literature and potential future areas of investigation . Non steroidal anti-inflammatory drugs and P35354 inhibitors as anti-cancer therapeutics : hypes , hopes and reality . Non-steroidal anti-inflammatory drugs ( NSAIDs ) and specific inhibitors of cyclooxygenase ( P36551 ) -2 , are therapeutic groups widely used for the treatment of pain , inflammation and fever . There is growing experimental and clinical evidence indicating NSAIDs and P35354 inhibitors also have anti-cancer activity . Epidemiological studies have shown that regular use of DB00945 and other NSAIDs reduces the risk of developing cancer , in particular of the colon . Molecular pathology studies have revealed that P35354 is expressed by cancer cells and cells of the tumor stroma during tumor progression and in response to chemotherapy or radiotherapy . Experimental studies have demonstrated that P35354 over expression promotes tumorigenesis , and that NSAIDs and P35354 inhibitors suppress tumorigenesis and tumor progression . Clinical trials have shown that NSAIDs and P35354 inhibitors suppress colon polyp formation and malignant progression in patients with familial adenomatous polyposis ( FAP ) syndrome . Recent advances in the understanding of the cellular and molecular mechanisms of the anti-cancer effects of NSAIDs and P35354 inhibitors have demonstrated that these drugs target both tumor cells and the tumor vasculature . The therapeutic benefits of P35354 inhibitors in the treatment of human cancer in combination with chemotherapy or radiotherapy are currently being tested in clinical trials . In this article we will review recent advances in the understanding of the anti-tumor mechanisms of these drugs and discuss their potential application in clinical oncology . 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 . Safe full-dose one-step nabumetone challenge in patients with nonsteroidal anti-inflammatory drug hypersensitivity . DB00945 and all nonsteroidal anti-inflammatory drugs ( NSAIDs ) are a chemically heterogeneous group of compounds that share the ability to inhibit the enzyme cyclooxygenase ( P36551 ) . This inhibitory effect , especially of P23219 , is suggested as the mechanism underlying NSAID-induced hypersensitivity reactions . In this study , we evaluated the safety and convenience of a single full-dose challenge with nabumetone , a selective P35354 inhibitor , in patients with hypersensitivity to nonselective NSAIDs ( ns-NSAIDs ) . Twenty-four subjects with a history of hypersensitivity reactions to at least two different ns-NSAIDs on two different occasions were enrolled in the study . The patients were otherwise healthy and did not suffer from NSAID- or aspirin-induced asthma or urticaria . All subjects were orally challenged by a single full dose ( 1000 mg ) of nabumetone , monitored closely in the hospital for the next 4 hours and contacted by telephone the next morning and 3-12 months afterward . Twenty-two patients tolerated nabumetone without any reaction during and after the challenge . One patient had a single urticarial lesion and one patient reported mild pruritus without objective signs , both of which resolved spontaneously . Thirteen patients , including the patient who responded with pruritus to the challenge , used nabumetone on several occasions during the follow-up period without any adverse reaction . Our study shows that in patients with a history of aspirin- and ns-NSAID-induced hypersensitivity reaction , a rapid one-step challenge with nabumetone was well tolerated . These initial data support the possibility that a single full dose of nabumetone can be tried as a safe alternative in most patients with a hypersensitivity reaction to ns-NSAIDs . The epidemiology of prostate cancer -- with a focus on nonsteroidal anti-inflammatory drugs . DB00945 and other NSAIDs have a potential role in the primary and secondary prevention of many common diseases associated with aging , including the top two causes of mortality in the United States-cardiovascular disease and cancer . These agents may be beneficial in the management of Alzheimer 's disease,other forms of dementia , and Parkinson's. disease . Because men with prostate cancer or precancer are likely to present with coexisting conditions that would be affected by systemic aspirin , NSAID , or other P35354 inhibitor therapies , it is important to consider any possible preventive studies or future clinical recommendations of aspirin or NSAIDs for prostate cancer within the context of these comorbid conditions . DB00945 or nonaspirin NSAIDs may be appropriate prevention therapy for patients at high risk of prostate cancer , myocardial infarction , Parkinson 's disease , Alzheimer 's disease , lung cancer , or colorectal cancer , but low risk for gastrointestinal complications or stroke . Further quantitative comparative studies of the risks and benefits of these common comorbidities in older Americans , with special attention to dose and duration parameters , are warranted . Activation of interleukin-32 pro-inflammatory pathway in response to influenza A virus infection . BACKGROUND : Interleukin ( IL ) -32 is a recently described pro-inflammatory cytokine that has been reported to be induced by bacteria treatment in culture cells . Little is known about P24001 production by exogenous pathogens infection in human individuals . METHODS AND FINDINGS : In this study , we found that P24001 level was increased by 58.2 % in the serum samples from a cohort of 108 patients infected by influenza A virus comparing to that of 115 healthy individuals . Another pro-inflammatory factor cyclooxygenase ( P36551 ) -2-associated prostaglandin E2 was also upregulated by 2.7-fold . Expression of P24001 in influenza A virus infected A549 human lung epithelial cells was blocked by either selective P35354 inhibitor NS398 or DB00945 , a known anti-inflammatory drug , indicating P24001 was induced through P35354 in the inflammatory cascade . Interestingly , we found that P35354 -associate PGE(2) production activated by influenza virus infection was significantly suppressed by over-expression of P24001 but increased by P24001 -specific siRNA , suggesting there was a feedback mechanism between P24001 and P35354 . CONCLUSIONS : P24001 is induced by influenza A virus infection via P35354 in the inflammatory cascade . Our results provide that P24001 is a potential target for anti-inflammatory medicine screening . 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 . 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 . 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 . Platelet thromboxane ( 11-dehydro-Thromboxane B2 ) and aspirin response in patients with diabetes and coronary artery disease . DB00945 ( ASA ) irreversibly inhibits platelet cyclooxygenase-1 ( P23219 ) leading to decreased thromboxane-mediated platelet activation . The effect of ASA ingestion on thromboxane generation was evaluated in patients with diabetes ( DM ) and cardiovascular disease . Thromboxane inhibition was assessed by measuring the urinary excretion of 11-dehydro-thromboxane B2 ( 11dhTxB2 ) , a stable metabolite of thromboxane A2 . The mean baseline urinary 11dhTxB2 of DM was 69.6 % higher than healthy controls ( P = 0.024 ) : female subjects ( DM and controls ) had 50.9 % higher baseline 11dhTxB2 than males ( P = 0.0004 ) , while age or disease duration had no influence . Daily ASA ingestion inhibited urinary 11dhTxB2 in both DM ( 71.7 % ) and controls ( 75.1 % , P < 0.0001 ) . Using a pre-established cut-off of 1500 pg/mg of urinary 11dhTxB2 , there were twice as many ASA poor responders ( ASA " resistant " ) in DM than in controls ( 14.8 % and 8.4 % , respectively ) . The rate of ASA poor responders in two populations of acute coronary syndrome ( ACS ) patients was 28.6 and 28.7 % , in spite of a significant ( 81.6 % ) inhibition of urinary 11dhTxB2 ( P < 0.0001 ) . Both baseline 11dhTxB2 levels and rate of poor ASA responders were significantly higher in DM and ACS compared to controls . Underlying systemic oxidative inflammation may maintain platelet function in atherosclerotic cardiovascular disease irrespective of P23219 pathway inhibition and/or increase systemic generation of thromboxane from non-platelet sources . DB00945 insensitive eicosanoid biosynthesis in cardiovascular disease . Inhibition of platelet cyclooxygenase ( P36551 ) -1 is involved in aspirin cardioprotection observed in clinical trials , but , in some patients , aspirin is unable to protect from thrombotic complications . An incomplete suppression of platelet thromboxane ( TX ) A2 biosynthesis has been assumed to participate in the phenomenon of aspirin resistance , as a consequence of the following possible mechanisms : ( i ) P35354 expression in newly formed platelets ; ( ii ) pharmacodynamic interactions between aspirin and coadministered nonsteroidal antiinflammatory drugs ( e.g. ibuprofen ) ; ( iii ) expression of variant isoforms of P23219 with reduced sensitivity to irreversible inactivation at Ser529 . Furthermore , aspirin failure may be due to enhanced formation of vasoactive and/or proaggregatory eicosanoids despite an almost complete suppression of platelet TXA2 biosynthesis by aspirin . Thus , in a subset of patients with unstable angina treated with low-dose aspirin , to almost completely block platelet P23219 activity , enhanced TXA2 biosynthesis in vivo has been demonstrated , presumably through an increased generation of P35354 -dependent PGH2 in plaque monocytes/macrophages or activated vascular cells . The concomitant increased formation of 8-iso-PGF2alpha , one of the most abundant F2-isoprostanes in humans , generated by free-radical catalyzed arachidonate peroxidation , may be involved in aspirin resistance because of its capacity to induce platelet and vascular smooth muscle cell activation through the interaction with thromboxane receptors ( TPs ) . Finally , enhanced production of vasoactive cysteinyl leukotrienes ( cys-LTs ) occurs in unstable angina despite conventional antithrombotic and antianginal treatment . The use of selective pharmacological tools ( i.e. highly selective P35354 inhibitors , TP antagonists , cys-LT inhibitors and antagonists ) will allow to ascertain the contribution of aspirin insensitive eicosanoid biosynthesis in aspirin cardioprotective failure . DB00945 induction of apoptosis in esophageal cancer : a potential for chemoprevention . The potential use of non-steroidal anti-inflammatory drugs ( NSAIDs ) in the prevention of gastrointestinal cancers has been highlighted recently . However , it is not known whether NSAIDs could also be useful for preventing esophageal cancer , although regular users of these drugs appear to have a decreased incidence of esophageal cancer . Therefore , we examined the effect of aspirin on growth and apoptosis in 10 esophageal cancer cell lines as well as the expression and modulation of its target enzymes , cyclooxygenases ( COXs ) , and their product prostaglandin E2 . Growth inhibition of these cells by aspirin was dose- and time-dependent and associated with the induction of apoptosis . P23219 and P35354 were expressed in 7 of the 10 cell lines . Bile acids could induce P35354 expression in six of eight cell lines tested , which was correlated with prostaglandin E2 production , and aspirin could inhibit P35354 enzymatic activity even after bile acid stimulation but was unable to change the P35354 protein level in these cell lines . Down-regulation of bcl-2 by aspirin was found in the two cell lines tested . These results suggest that induction of apoptosis by aspirin may be a mechanism by which it can intervene in esophageal carcinogenesis and may be indicative of the potential of NSAIDs as chemopreventive agents in esophageal cancer . 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 . 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 . 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 . Role of Tyr348 in Tyr385 radical dynamics and cyclooxygenase inhibitor interactions in prostaglandin H synthase-2 . Both prostaglandin H synthase ( PGHS ) isoforms utilize a radical at Tyr385 to abstract a hydrogen atom from arachidonic acid , initializing prostaglandin synthesis . A Tyr348-Tyr385 hydrogen bond appears to be conserved in both isoforms ; this hydrogen bonding has the potential to modulate the positioning and reactivity of the Tyr385 side chain . The EPR signal from the Tyr385 radical undergoes a time-dependent transition from a wide doublet to a wide singlet species in both isoforms . In P35354 , this transition results from radical migration from Tyr385 to Tyr504 . Localization of the radical to Tyr385 in the recombinant human P35354 Y504F mutant was exploited in examining the effects of blocking Tyr385 hydrogen bonding by introduction of a further Y348F mutation . Cyclooxygenase and peroxidase activities were found to be maintained in the Y348F/Y504F mutant , but the Tyr385 radical was formed more slowly and had greater rotational freedom , as evidenced by observation of a transition from an initial wide doublet species to a narrow singlet species , a transition not seen in the parent Y504F mutant . The effect of disrupting Tyr385 hydrogen bonding on the cyclooxygenase active site structure was probed by examination of cyclooxygenase inhibitor kinetics . DB00945 treatment eliminated all oxygenase activity in the Y348F/Y504F double mutant , with no indication of the lipoxygenase activity observed in aspirin-treated wild-type P35354 . Introduction of the Y348F mutation also strengthened the time-dependent inhibitory action of nimesulide . These results suggest that removal of Tyr348-Tyr385 hydrogen bonding in P35354 allows greater conformational flexibility in the cyclooxygenase active site , resulting in altered interactions with inhibitors and altered Tyr385 radical behavior . DB00945 and stroke prevention . According to meta-analyses aspirin provides a relative reduction in the rate of major vascular events of 19 % in patients with arterial disease in general , whereas for patients with ischaemic cerebrovascular disease this reduction is only 13 % . The discrepancy may well result from pathophysiological differences and not from a play of chance . There is no proven difference in efficacy according to dose . The evidence for this equivalence is most compelling in the range between 75 and 1300 mg daily , but still fairly convincing for doses between 30 and 50 mg . In contrast , side effects are clearly more frequent as the dose is higher . Other antiplatelet agents ( sulfinpyrazone , ticlopidine , clopidogrel , dipyridamole , orally administered IIb/IIIa inhibitors ) have no clear advantages over aspirin and in some cases definite disadvantages ; the combination of aspirin and dipyridamole may be more efficacious than aspirin alone , but the evidence hinges on a single trial . If recurrent TIAs occur under treatment with aspirin , the rational response is not to change to a different antiplatelet agent , but to review the diagnosis and consider causes other than artery-to-artery embolism . Platelet aggregation can probably still occur despite complete acetylation of platelets , via pathways other than P23219 inhibition , but in vitro aggregation tests are an unreliable measure . Leukocyte lipid body formation and eicosanoid generation : cyclooxygenase-independent inhibition by aspirin . Lipid bodies , cytoplasmic inclusions that develop in cells associated with inflammation , are inducible structures that might participate in generating inflammatory eicosanoids . Cis-unsaturated fatty acids ( arachidonic and oleic acids ) rapidly induced lipid body formation in leukocytes , and this lipid body induction was inhibited by aspirin and nonsteroidal antiinflammatory drugs ( NSAIDs ) . Several findings indicates that the inhibitory effect of aspirin and NSAIDs on lipid body formation was independent of cyclooxygenase ( P36551 ) inhibition . First , the non- P36551 inhibitor , sodium salicylate , was as potent as aspirin in inhibiting lipid body formation elicited by cis-fatty acids . Second , cis-fatty acid-induced lipid body formation was not impaired in macrophages from P23219 or P35354 genetically deficient mice . Finally , NSAIDs inhibited arachidonic acid-induced lipid body formation likewise in macrophages from wild-type and P23219 - and P35354 -deficient mice . An enhanced capacity to generate eicosanoids developed after 1 hr concordantly with cis-fatty acid-induced lipid body formation . Arachidonic and oleic acid-induced lipid body numbers correlated with the enhanced levels of leukotrienes B4 and C4 and prostaglandin E2 produced after submaximal calcium ionophore stimulation . DB00945 and NSAIDs inhibited both induced lipid body formation and the enhanced capacity for forming leukotrienes as well as prostaglandins . Our studies indicate that lipid body formation is an inducible early response in leukocytes that correlates with enhanced eicosanoid synthesis . DB00945 and NSAIDs , independent of P36551 inhibition , inhibit cis-fatty acid-induced lipid body formation in leukocytes and in concert inhibit the enhanced synthesis of leukotrienes and prostaglandins . DB00945 treatment influences platelet-related inflammatory biomarkers in healthy individuals but not in acute stroke patients . OBJECTIVES : Platelet-leukocyte aggregation is believed to contribute to acute thrombotic events . While the effect of aspirin on platelet-to-platelet aggregation is well established , the impact of the drug on pro-inflammatory platelet function remains equivocal . Thus we investigated the effect of aspirin on selected platelet-related inflammatory biomarkers in both acute ischaemic stroke patients and healthy volunteers . METHODS : Using five-colour flow cytometry the platelet surface expression of CD62P and P29965 and subpopulations of leukocyte-platelet aggregates were assessed in 63 acute stroke patients and 40 healthy volunteers at baseline and after a 10-day period of aspirin intake at a daily dose of 150 mg . Simultaneously the plasma levels of soluble CD62P and P29965 , serum level of TxB(2) , and whole blood impedance platelet aggregation under arachidonic acid ( AA ) stimulation were investigated . RESULTS : No differences in values of studied platelet-related inflammatory biomarkers in both resting platelets and those activated with TRAP after 10-day treatment with aspirin were confirmed in stroke subjects . In healthy individuals the resting platelet expression of CD62P , plasma level of soluble CD62P and percentage of circulating monocyte-platelet aggregates were lower after the aspirin intake period ( P=0.009 ; P=0.04 ; P=0.004 , respectively ) . In both studied groups serum level of TxB(2) and platelet aggregation under AA stimulation were lower than before treatment ( P < 0.001 ) . CONCLUSION : Despite effective inhibition of P23219 -dependent platelet aggregation , aspirin does not influence the platelet α-granule-derived inflammatory mediators and monocyte-platelet aggregation in acute stroke subjects , although it does in healthy individuals . 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 . DB00945 sensitivity and desensitization for asthma and sinusitis . NSAIDs-including aspirin ( ASA ) -that inhibit cyclooxygenase ( P36551 ) -1 induce nonallergic hypersensitivity reactions consisting of attacks of rhinitis and asthma . Such reactions occur exclusively in a subset of asthmatic patients who also have underlying nasal polyps and chronic hyperplastic eosinophilic sinusitis . We now refer to their underlying inflammatory disease of the entire respiratory tract as aspirin-exacerbated respiratory disease . This review focuses on descriptions of these patients ; methods available to diagnose ASA-exacerbated respiratory disease ; the unique ability of all NSAIDs that inhibit P23219 to cross-react with ASA ; lack of cross-reactivity with selective P35354 inhibitors ; an update on pathogenesis ; and current thoughts about treatment , including ASA desensitization and daily ingestion of ASA itself . Prostaglandin G/H synthase-2 is required for maximal formation of osteoclast-like cells in culture . We examined the effect on osteoclast formation of disrupting the prostaglandin G/H synthase genes P23219 and-2 . Prostaglandin E(2) ( PGE(2) ) production was significantly reduced in marrow cultures from mice lacking P35354 ( P35354 (-/-) ) compared with wild-type ( P35354 (+/+) ) cultures . Osteoclast formation , whether stimulated by 1,25-dihydroxyvitamin D(3) ( 1,25-D ) or by parathyroid hormone ( PTH ) , was reduced by 60-70 % in P35354 (-/-) cultures relative to wild-type cultures , an effect that could be reversed by providing exogenous PGE(2) . Cultures from heterozygous mice showed an intermediate response . PGHS inhibitors caused a similar drop in osteoclast formation in wild-type cultures . Co-culture experiments showed that supporting osteoblasts , rather than osteoclast precursors , accounted for the blunted response to 1,25-D and PTH . This lack of response appeared to result from reduced expression of Q9Y6Q6 ligand ( O14788 ) in osteoblasts . We cultured spleen cells with exogenous O14788 and found that osteoclast formation was 50 % lower in P35354 (-/-) than in wild-type cultures , apparently because the former cells expressed high levels of GM- P04141 . Injection of PTH above the calvaria caused hypercalcemia in wild-type but not P35354 (-/-) mice . Histological examination of bone from 5-week-old P35354 (-/-) mice revealed no abnormalities . Mice lacking P23219 were similar to wild-type mice in all of these parameters . These data suggest that P35354 is not necessary for wild-type bone development but plays a critical role in bone resorption stimulated by 1,25-D and PTH . 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 . [ 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 . Cyclooxygenase-independent chemoprevention with an aspirin derivative in a rat model of colonic adenocarcinoma . DB00945 decreases the risk of colorectal cancer , reportedly through suppression of cyclooxygenase ( P36551 ) activity . Using a rat model of colonic adenocarcinoma , we compared the chemopreventative effects of aspirin versus a nitric oxide-releasing derivative ( O43763 -4016 ) which does not inhibit P36551 . Beginning six weeks after intracolonic administration of trinitrobenzene sulfonic acid , the rats were given azoxymethane weekly ( 15 mg/kg i.p. ) for 4 weeks . Over the same 4-week period , the rats were treated daily with vehicle , aspirin ( 10 mg/kg ) or O43763 -4016 ( equimolar dose ) . Six weeks later , the number of aberrant crypt foci ( an early preneoplastic lesion ) were blindly counted by light microscopy . Effects of aspirin vs. O43763 -4016 on P23219 and P35354 activity were compared , as was their analgesic activity . Rats receiving vehicle developed a mean of 856 +/- 260 aberrant crypt foci in the colon . DB00945 reduced the number of aberrant crypt foci by 64 % , while O43763 -4016 produced an 85 % reduction . DB00945 , but not O43763 -4016 , markedly suppressed systemic P23219 and P35354 activity , and colonic prostaglandin synthesis . Despite not inhibiting P36551 , O43763 -4016 exhibited comparable analgesic activity to aspirin . These results demonstrate that O43763 -4016 , a nitric oxide-releasing aspirin derivative , exhibited superior chemopreventative effects to aspirin in this model of colon cancer . This effect occurred independent of inhibition of P23219 or P35354 . New perspectives on aspirin and the endogenous control of acute inflammatory resolution . DB00945 is unique among the nonsteroidal anti-inflammatory drugs in that it has both anti-inflammatory as well as cardio-protective properties . The cardio-protective properties arise form its judicious inhibition of platelet-derived thromboxane A2 over prostacyclin , while its anti-inflammatory effects of aspirin stem from its well-established inhibition of prostaglandin ( PG ) synthesis within inflamed tissues . Thus aspirin and the other NSAIDs have popularised the notion of inhibiting PG biosynthesis as a common anti-inflammatory strategy based on the erroneous premise that all eicosanoids are generally detrimental to inflammation . However , our fascination with aspirin has shown a more affable side to lipid mediators based on our increasing interest in the endogenous control of acute inflammation and in factors that mediate its resolution . Epi-lipoxins ( epi-LXs ) , for instance , are produced from aspirin 's acetylation of inducible cyclooxygenase 2 ( P35354 ) and together with Resolvins represent an increasingly important family of immuno-regulatory and potentially cardio-protective lipid mediators . DB00945 is beginning to teach us what nature knew all along -- that not all lipid mediators are bad . It seems that while some eicosanoids are pathogenic in a variety of diseases , others are unarguable protective . In this review we will re-count aspirin 's colorful history , discuss its traditional mode of action and the controversies associated therewith , as well as highlight some of the new pathways in inflammation and the cardiovascular systems that aspirin has recently revealed . DB00945 -induced asthma : advances in pathogenesis , diagnosis , and management . In some asthmatic individuals , aspirin and other nonsteroidal anti-inflammatory drugs ( NSAIDs ) that inhibit cyclooxygen-ase 1 ( P23219 ) exacerbate the condition . This distinct clinical syndrome , called aspirin-induced asthma ( AIA ) , is characterized by an eosinophilic rhinosinusitis , nasal polyposis , aspirin sensitivity , and asthma . There is no in vitro test for the disorder , and diagnosis can be established only by provocation challenges with aspirin or NSAIDs . Recent major advances in the molecular biology of eicosanoids , exemplified by the cloning of 2 cysteinyl leukotriene receptors and the discovery of a whole family of cyclooxygenase enzymes , offer new insights into mechanisms operating in AIA . The disease runs a protracted course even if P23219 inhibitors are avoided , and the course is often severe , many patients requiring systemic corticosteroids to control their sinusitis and asthma . DB00945 and NSAIDs should be avoided , but highly specific P35354 inhibitors , known as coxibs , are well tolerated and can be safely used . DB00945 desensitization , followed by daily aspirin treatment , is a valuable therapeutic option in most patients with AIA , particularly those with recurrent nasal polyposis or overdependence on systemic corticosteroids . [ 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 .

MH_train_1155

MH_train_1155

MH_train_1155

interacts_with DB00620?

multiple_choice

Superagonistic action of 14-epi-analogs of 1,25-dihydroxyvitamin D explained by vitamin D receptor-coactivator interaction . Two 14-epi-analogs of 1,25-dihydroxyvitamin D3 [ 1,25-(OH)(2)D(3) ] , 19-nor-14-epi-23-yne-1,25-(OH)2D3 ( TX522 ) and 19-nor-14,20-bisepi-23-yne-1,25-(OH)2D3 ( TX527 ) , show enhanced antiproliferative ( at least 10-fold ) and markedly lower calcemic effects both in vitro and in vivo , compared with 1,25-(OH)2D3 . This study aimed to evaluate their superagonistic effect at the level of interaction between the Vitamin D receptor ( P11473 ) and coactivators . Mammalian two-hybrid assays with VP16-fused P11473 and GAL4-DNA-binding-domain-fused steroid receptor coactivator 1 ( Q15788 ) , transcriptional intermediary factor 2 ( Tif2 ) , or Q15648 showed the 14-epi-analogs to be more potent inducers of P11473 -coactivator interactions than 1,25-(OH)2D3 ( up to 16- and 20-fold stronger induction of P11473 - Q15788 interaction for TX522 and TX527 at 10(-10) M ) . Similar assays in which metabolism of 1,25-(OH)2D3 was blocked with VID400 , a selective inhibitor of the 1,25-(OH)2D3-metabolizing enzyme Q07973 , showed that the enhanced potency of these analogs in establishing P11473 -coactivator interactions can only partially be accounted for by their increased resistance to metabolic degradation . Crystallization of TX522 complexed to the ligand binding domain of the human P11473 demonstrated that the epi-configuration of C14 caused the CD ring of the ligand to shift by 0.5 angstroms , thereby bringing the C12 atom into closer contact with Val300 . Moreover , C22 of TX522 made an additional contact with the CD1 atom of Ile268 because of the rigidity of the triple bond-containing side chain . The position and conformation of the activation helix H12 of P11473 was strictly maintained . In conclusion , this study provides deeper insight into the docking of TX522 in the P18428 and shows that stronger P11473 -coactivator interactions underlie the superagonistic activity of the two 14-epi-analogs . 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 . DB03496 induces mitochondrial-mediated apoptosis in murine glioma GL261 cells via release of cytochrome c and apoptosis inducing factor . Glioblastoma ( GBM ) remains one of the most challenging solid cancers to treat due to its highly proliferative , angiogenic and invasive nature . The small molecule CDK inhibitor , flavopiridol , has demonstrated antitumor activity in human xenograft models and is currently in clinical trials showing efficacy in patients with advanced disease . We have developed an experimental animal model using the murine glioma GL261 cells as a novel in vivo system to screen potential therapeutic agents for GBM . Results of in vitro testing demonstrate that flavopiridol has several relevant clinical characteristics such as its ability to : 1. inhibit cell growth ; 2. inhibit cell migration ; 3. decrease expression of cyclin D1 , P11802 and P38936 ; 4. induce apoptosis in cells with high levels of p27 expression ; and 5. decrease the expression of the anti-apoptotic protein Bcl-2 . The mechanism by which flavopiridol induces apoptosis is mitochondrial-mediated . We demonstrate by electron microscopy and immunohistochemistry that drug treatment induces mitochondrial damage that was accompanied by the release of cytochrome c into the cytosol together with the translocation of apoptosis inducing factor ( O95831 ) into the nucleus . This finding in murine glioma cells differs from the mechanism of flavopiridolinduced cell death reported by us for human glioma cells ( Alonso et al. , Mol Cancer Ther 2003 ; 2:139 ) where drug treatment induced a caspase- and cytochrome c-independent pathway in the absence of detectable damage to mitochondria . In apoptotic human glioma cells only translocation of O95831 into the nucleus occurred . Thus , the same drug kills different types of glioma cells by different mitochondrial-dependent pathways . Differential P24522 , p21CIP1/ P38936 , Q8WXI8 -1 and topoisomerase II gene induction and secondary DNA fragmentation after camptothecin-induced DNA damage in two mutant p53 human colon cancer cell lines . DB04690 ( CPT ) traps covalent P11387 -linked DNA single-strand breaks ( cleavable complexes ) . To determine the differences in DNA damage signalling leading to differential sensitivity to CPT , two human colon cancer cell lines , SW620 and KM12 , with nonfunctional p53 and the same level of topoisomerase I cleavable complex formation but differential sensitivity to CPT ( Cancer Res. 56:4430-7 ; 1996 ) were studied . The levels of mRNA expression of DNA damage-inducible or death-related genes were measured at different times after CPT treatment . KM12 cells exhibited 3-fold higher basal levels of BCL-2 mRNA . Consistently , secondary DNA fragmentation , quantitated using a filter elution assay , was detected 24 h later and was 2-4-fold lower in KM12 cells than in SW620 cells . No induction of Q07812 was detected in either cell line . Consistent with the absence of functional p53 , p21CIP1/ P38936 and P24522 genes were not induced within the first 24 h . However , in SW620 cells , both mRNA levels were increased more than 10-fold at 48 h . The BCL-2-related gene Q8WXI8 -1 and topoisomerase II mRNA were induced at 24 h , and topoisomerase I mRNA levels increased 3-fold at 48 h , only in SW620 cells . We conclude that cellular response to CPT-induced DNA damage can involve p53-independent pathways leading to the induction of p53-effector genes . Induction of these genes at the onset of apoptosis is associated with CPT sensitivity . Adipocyte-specific reduction of phosphodiesterase 3B gene expression and its restoration by DB04971 in the obese , diabetic KKAy mouse . OBJECTIVE : Phosphodiesterase ( PDE ) 3B is a key enzyme involved in the anti-lipolytic action of insulin in adipocytes . Q13370 activation results in a reduced output of free fatty acids ( FFA ) , whereas elevated serum FFA is known to cause insulin resistance . We have recently reported that reduced Q13370 gene expression is restored by treatment with pioglitazone , in the adipose tissues of obese , insulin-resistant diabetic KKAy mice . To determine whether the altered Q13370 gene expression is specific for adipocytes , the expression of this gene in liver and epididymal fat tissues of KKAy mice was examined . The effect of DB04971 , another peroxisome proliferator-activated receptor ( Q07869 )gamma ligand , which is different from thiazolidinedione , was also examined . METHODS : Q13370 mRNA and protein were quantified by an RNase protection assay and Western blotting respectively . Membrane-bound PDE activities were also measured . RESULTS : In adipose tissues of KKAy mice , Q13370 mRNA , protein and membrane-bound PDE activity were reduced to 47 % , 57 % and 51 % respectively relative to those in C57BL/6J control mice . DB04971 increased Q13370 mRNA , protein and membrane-bound PDE activity by 2.2- , 1.6- and 1.7-fold respectively over those of untreated KKAy mice . In the liver , Q13370 gene expression remained unchanged in KKAy mice , and was not affected by DB04971 . DB04971 reduced the elevated levels of serum insulin , glucose , FFA and triglyceride in KKAy mice . CONCLUSIONS : Q13370 gene expression was specifically reduced in the adipose tissues of KKAy mice . DB04971 restored this reduced gene expression with an accompanying improvement in elevated serum FFA and insulin resistance . Regulation of the human P38936 (waf1/cip1) gene promoter via multiple binding sites for p53 and the vitamin D3 receptor . The main regulator of the human tumor suppresser gene P38936 (waf1/cip1) is the transcription factor p53 , but more recently it has been suggested to be a primary anti-proliferative target for the nuclear receptor P11473 in the presence of its ligand 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) . To identify P11473 responding regions , we analyzed 20 overlapping regions covering the first 7.1 kb of the P38936 (waf1/cip1) promoter in MCF-7 human breast cancer cells using chromatin immuno-precipitation assays ( ChIP ) with antibodies against p53 and P11473 . We confirmed two known p53 binding regions at approximate positions -1400 and -2300 and identified a novel site at position -4500 . In addition , we found three P11473 -associated promoter regions at positions -2300 , -4500 and -6900 , i.e. two regions showed binding for both p53 and P11473 . In silico screening and in vitro binding assays using recombinant and in vitro translated proteins identified five p53 binding sites within the three p53-positive promoter regions and also five DB00136 response elements within the three P11473 -positive regions . Reporter gene assays confirmed the expected responsiveness of the respective promoter regions to the p53 inducer 5-fluorouracil and DB00136 . Moreover , re-ChIP assays confirmed the functionality of the three DB00136 -reponsive promoter regions by monitoring simultaneous occupancy of P11473 with the co-activator proteins CBP , Q15788 and Q15648 . Taken together , we demonstrated that the human P38936 ( ( waf1/cip1 ) ) gene is a primary DB00136 -responding gene with at least three P11473 binding promoter regions , in two of which also p53 co-localizes . Targeted cytotoxic analog of luteinizing hormone-releasing hormone ( P01148 ) , AEZS-108 ( AN-152 ) , inhibits the growth of DU-145 human castration-resistant prostate cancer in vivo and in vitro through elevating P38936 and ROS levels . Management of castration-resistant prostate cancer ( CRPC ) is challenging due to lack of efficacious therapy . DB00044 -releasing hormone analogs appear to act directly on cells based on the P01148 receptors on human prostate adenocarcinoma cells . We explored anticancer activity of a cytotoxic analog of P01148 , AEZS-108 consisting of P01148 agonist linked to doxorubicin . Nude mice bearing DU-145 tumors were used to compare antitumor effects of AEZS-108 with its individual constituents or their unconjugated combination . The tumor growth inhibition of conjugate was greatest among treatment groups ( 90.5 % inhibition vs. 41 % by [D-Lys(6)] P01148 +DOX ) . The presence of P01148 receptors on DU-145 cells was confirmed by immunocytochemistry . In vitro , AEZS-108 significantly inhibited cell proliferation ( 61.2 % inhibition ) and elevated apoptosis rates ( by 46 % ) . By the detection of the inherent doxorubicin fluorescence , unconjugated doxorubicin was seen in the nucleus ; the conjugate was perinuclear and at cell membrane . Autophagy , visualized by GFP-tagged p62 reporter , was increased by AEZS-108 ( 7.9-fold vs. 5.3-fold by DOX+[D-Lys(6)] P01148 . AEZS-108 more effectively increased reactive oxygen species ( ROS , 2-fold vs. 1.4-fold by DOX+[D-Lys(6)] P01148 ) and levels of the apoptotic regulator P38936 in vivo and in vitro . We demonstrate robust inhibitory effects of the targeted cytotoxic P01148 analog AEZS-108 on P22888 positive castration-resistant prostate cancer cells . Q07869 ( PPARalpha ) activators induce hepatic farnesyl diphosphate synthase gene expression in rodents . Fibrates are hypolipidemic drugs that exert multiple effects on lipid metabolism by activating peroxisome proliferator-activated receptor alpha ( PPARalpha ) and modulating the expression of many target genes . In order to investigate the link between PPARalpha and cholesterol synthesis , we analysed the effect of fibrates on expression of the farnesyl diphosphate synthase ( P14324 ) gene , known to be regulated by sterol regulatory element-binding proteins ( SREBPs ) , in conjunction with P04035 . In wild-type mice , both fenofibrate and WY 14,643 induced P14324 gene expression , an effect impaired in PPARalpha-null mice . A three-fold induction was observed in ciprofibrate-treated rat hepatocytes , in primary culture . This effect was decreased in presence of 5,6-dichlorobenzimidazole riboside ( DRB ) and cycloheximide ( CHX ) , transcription and translation inhibitors , respectively . Acyl- DB01992 oxidase ( Q15067 ) , a bona fide PPARalpha target gene , was induced by ciprofibrate but slower and more strongly than P14324 . In addition , induction of P14324 gene expression was abolished in the presence of 25-hydroxycholesterol ( 25-OH Chol ) . Thus , activation of PPARalpha by fibrates induced P14324 gene expression in both hepatocytes in culture and in mouse liver . This effect is likely to be dependent on cellular sterol level , possibly through SREBP-mediated transcriptional activation . DB04630 induces interleukin-18 through endothelin-1 , angiotensin II , Rho/Rho-kinase , and PPARs in cardiomyocytes . DB04630 ( Aldo ) is recognized as an important risk factor for cardiovascular diseases . Q14116 induces myocardial hypertrophy , loss of contractility of cardiomyocytes , and apoptosis leading myocardial dysfunction . However , so far , there have been few reports concerning the interaction between Aldo and Q14116 . The present study examined the effects and mechanisms of Aldo on Q14116 expression and the roles of peroxisome proliferator-activated receptor ( Q07869 ) agonists in rat cardiomyocytes . We used cultured rat neonatal cardiomyocytes stimulated with Aldo to measure Q14116 mRNA and protein expression , Rho-kinase , and NF-kappaB activity . We also investigated the effects of Q07869 agonists on these actions . Aldo , endothelin-1 ( ET-1 ) , and angiotensin II ( P03950 II ) increased Q14116 mRNA and protein expression . P08235 antagonists , endothelin A receptor antagonist , and P03950 II receptor antagonist inhibited Aldo-induced Q14116 expression . Aldo induced ET-1 and P03950 II production in cultured media . Moreover , Rho/Rho-kinase inhibitor and statin inhibited Aldo-induced Q14116 expression . On the other hand , Aldo upregulated the activities of Rho-kinase and NF-kappaB . Q07869 agonists attenuated the Aldo-induced Q14116 expression and NF-kappaB activity but not the Rho-kinase activity . Our findings indicate that Aldo induces Q14116 expression through a mechanism that involves , at a minimum , ET-1 and P03950 II acting via the Rho/Rho-kinase and Q07869 /NF-kappaB pathway . The induction of Q14116 in cardiomyocytes by Aldo , ET-1 , and P03950 II might , therefore , cause a deterioration of the cardiac function in an autocrine and paracrine fashion . The inhibition of the Q14116 expression by Q07869 agonists might be one of the mechanisms whereby the beneficial cardiovascular effects are exerted . 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 . beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells . Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention , the molecular mechanisms underlying such chemopreventive activity are not clear . Since peroxisome proliferator-activated receptor gamma ( P37231 ) has been implicated in regulating breast cancer cell differentiation and apoptosis , the effects of beta-carotene on the P37231 -mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study . The results demonstrated that beta-carotene significantly increased P37231 mRNA and protein levels in time-dependent manner . In addition , beta-carotene increased the cyclin-dependent kinase inhibitor P38936 ( P38936 /CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression . DB07863 ( GW9662 ) , an irreversible P37231 antagonist , partly attenuated the cell death caused by beta-carotene . Further , reactive oxygen species ( ROS ) production was induced by beta-carotene , resulting in mitochondrial dysfunction and cytochrome C release . DB00143 ( DB00143 ) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene . In total , these observations suggest that the synergistic effect of P37231 expression and ROS production may account for beta-carotene-mediated anticancer activities . Optimal bone health management strategies in patients with prostate cancer . Bone health is affected in patients with prostate cancer , both by the disease and its treatment . Metastases to bone leads to pain , fractures , and spinal cord compression ; bone loss due to androgen deprivation therapy ( ADT ) leads to osteoporosis and its complications . Both these scenarios are a major cause of morbidity and adversely affect the quality of life of these patients . Maintaining an optimum bone health throughout the natural course of prostate cancer is an important aspect in the management of this disease . An understanding of the complex interplay between osteoclasts , osteoblasts , receptor activator of nuclear factor κB ( Q9Y6Q6 ) , and various other tyrosine kinases involved in the pathophysiology of bone metastases is essential . DB00399 ( ZA ) , an intravenously administered bisphosphonate , and DB06643 , a subcutaneously administered inhibitor of nuclear factor B ligand ( O14788 ) , have already been approved by Food and Drug Administration ( FDA ) for their use in treatment of bone metastases . This article discusses the pathophysiology of bone metastases and bone loss due to ADT in prostate cancer , role of biomarkers , newer modalities of imaging , World Health Organization ( WHO ) /FRAX nomogram in evaluation of these patients , utility of currently available drugs and evidence supporting their use , and newer therapeutic agents like alpha-emitting Radium-223 , endothelin-A receptor antagonists ( Atrasentan and Zibotentan ) and the proto-oncogene tyrosine-protein kinase ( P12931 ) inhibitor , Dasatinib . 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 . 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 . Differential role of two P11473 coactivators , Q15648 and Q9Y6Q9 , in keratinocyte proliferation and differentiation . Cell programs such as proliferation and differentiation involve the selective activation and repression of gene expression . The vitamin D receptor ( P11473 ) , through 1,25(OH)(2)D(3) , controls the proliferation and differentiation of keratinocytes . Previously , we have identified two P11473 binding coactivator complexes . In proliferating keratinocytes P11473 bound preferentially to the DRIP complex , whereas in differentiated keratinocytes the P12931 complex was preferred . We proposed that different coactivators are required for sequential gene regulation in the transition from proliferation to differentiation . Here we examined the roles of Q15648 and Q9Y6Q9 in this transition . Silencing of Q15648 and P11473 caused hyperproliferation of keratinocytes , demonstrated by increased XTT and BrdU incorporation . Q9Y6Q9 silencing , on the other hand , did not have an effect on proliferation . In contrast , Q9Y6Q9 as well as Q15648 and P11473 silencing blocked keratinocyte differentiation as shown by decreased expression of keratin 1 and filaggrin . These results are consistent with the differential localization of Q15648 and Q9Y6Q9 in skin . These results indicate that Q15648 is required for keratinocyte proliferation . Both Q15648 and Q9Y6Q9 are required for the keratinocyte differentiation . These results support the concept that the selective use of coactivators by P11473 underlies the selective regulation of gene expression in keratinocyte proliferation and differentiation .

MH_train_1156

MH_train_1156

MH_train_1156

interacts_with DB00991?

multiple_choice

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 . 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 . [ Association and interaction of AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 and P35354 genes on the risk of hypertension in Antioquian population ] . INTRODUCTION : Hypertension is a multifactorial disease influenced by genetic and environmental components , with its prevalence varying across ethnic groups . Manifold studies on blood pressure regulatory system genes have been carried out -such as the renin-angiotensin-aldosterone system , the sympathetic nervous system , endothelial factor , and sodium balance- , but the results yielded were inconsistent among populations . OBJECTIVES : To evaluate the effect of both variants in genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 P35354 , and the result of the individual ancestry component on hypertension and blood pressure levels among population in Antioquia . METHODS AND MATERIALS : 107 cases and 253 controls were genotyped for 12 variants on genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 y P35354 , and for 20 ancestry informative markers . The association of polymorphisms and their interactions , and the association of ancestral genetic composition with hypertension and blood pressure levels were examined . RESULTS : Genes P35612 , rs4852706 ( OR=3.0 ; p=0.023 ) ; P21728 , rs686 ( OR=0.38 ; p=0.012 ) and P07550 , rs1042718 ( OR=10.0 ; p=0.008 ) ; as well as genotypic combinations of P21728 and P30556 ; AGT and P35611 ; and P35611 to P20020 and P35354 were associated to hypertension . The Amerindian ancestry component was associated to some decrease in diastolic blood pressure . CONCLUSION : Variants on genes P35612 , P21728 , P07550 , P30556 , AGT , P35611 , P20020 and P35354 individually or interacting , are associated to hypertension . The Amerindian ancestry component has an effect on blood pressure . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS ) Opposite effects of tamoxifen on metabolic syndrome-induced bladder and prostate alterations : a role for Q99527 / Q99527 ? BACKGROUND : BPH and LUTS have been associated to obesity , hypogonadism , and metabolic syndrome ( MetS ) . MetS-induced prostate and bladder alterations , including inflammation and tissue remodeling , have been related to a low-testosterone and high-estrogen milieu . In addition to ERs , Q99527 / Q99527 is able to mediate several estrogenic non-genomic actions . METHODS : Supplementing a subgroup of MetS rabbits with tamoxifen , we analyzed the in vivo effects on MetS-induced prostate and bladder alterations . The effects of selective ER/ Q99527 ligands and Q99527 silencing on prostate inflammation were also studied in vitro using hBPH cells . RESULTS : ERα , ERβ , and PR expression was upregulated in MetS bladder , where tamoxifen decreased ERα and PR expression , further stimulating ERβ . In addition , tamoxifen-dosing decreased MetS-induced overexpression of inflammatory and tissue remodeling genes . In prostate , sex steroid receptors , pro-inflammatory and pro-fibrotic genes were upregulated in MetS . However , tamoxifen did not affect them and even increased P35354 . In hBPH cells , 17β-estradiol increased P10145 secretion , an effect blunted by co-treatment with Q99527 antagonist Q99943 but not by ER antagonist DB00947 , which further increased it . Q99527 agonist P55008 dose-dependently ( IC50 = 1.6 nM ) induced P10145 secretion . In vitro analysis demonstrated that Q99527 silencing reverted these stimulatory effects . CONCLUSIONS : Q99527 can be considered the main mediator of estrogen action in prostate , whereas in bladder the mechanism appears to rely on ERα , as indicated by in vivo experiments with tamoxifen dosing . Limiting the effects of the MetS-induced estrogen action via Q99527 could offer new perspectives in the management of BPH/LUTS , whereas tamoxifen dosing showed potential benefits in bladder . 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 . Role of cardiovascular aldosterone in hypertension . DB04630 plays an important role in the pathogenesis of cardiovascular disease . We have reported that aldosterone is synthesized in cardiovascular tissues and local aldosterone synthesis plays important roles for hypertension and cardiac hypertrophy . High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP . Plasma aldosterone concentrations and P06703 were decreased by high salt intake in SHRSP . DB04630 production , the expression of P19099 mRNA and angiotensin II receptor AT1R mRNA in blood vessels were significantly increased by high salt intake . These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the vascular tissue in SHRSP , which may contribute to the development of malignant hypertension in salt-loaded SHRSP . However , there are several reports of conflicting data . P08235 ( MR ) binding is tightly regulated by the enzyme 11beta-hydroxysteroid dehydrogenase type 2 ( 11beta-HSD2 ) which selectively metabolizes glucocorticoids to inactive metabolites , thus allowing for MR activation by aldosterone . We have reported that decreased 11beta-HSD2 in blood vessels in Dahl salt-sensitive ( DS ) rats , a model for salt-sensitive hypertension . Local aldosterone excess may play a significant role in the salt sensitivity and development of hypertension . High sodium intake decreased circulating rennin-angiotensin-aldosterone system and increased blood pressure and cardiac hypertrophy in DS rats , which were prevented by the treatment with a selective MR antagonist , eplerenone . DB00700 also improved endothelial nitric oxide synthase ( P29474 ) activity and P29474 mRNA expression in blood vessels in DS rats . These results further suggest that not only circulating aldosterone but also local aldosterone is of critical importance in the pathophysiology of cardiovascular disorders . Phase 1 clinical results with DB05465 ( MLN518 ) , a novel P36888 antagonist , in patients with acute myelogenous leukemia or high-risk myelodysplastic syndrome : safety , pharmacokinetics , and pharmacodynamics . Tandutinib ( MLN518/CT53518 ) is a novel quinazoline-based inhibitor of the type III receptor tyrosine kinases : P07333 -like tyrosine kinase 3 ( P36888 ) , platelet-derived growth factor receptor ( P09619 ) , and P10721 . Because of the correlation between P36888 internal tandem duplication ( ITD ) mutations and poor prognosis in acute myelogenous leukemia ( AML ) , we conducted a phase 1 trial of DB05465 in 40 patients with either AML or high-risk myelodysplastic syndrome ( P43034 ) . Tandutinib was given orally in doses ranging from 50 mg to 700 mg twice daily The principal dose-limiting toxicity ( DLT ) of DB05465 was reversible generalized muscular weakness , fatigue , or both , occurring at doses of 525 mg and 700 mg twice daily . Tandutinib 's pharmacokinetics were characterized by slow elimination , with achievement of steady-state plasma concentrations requiring greater than 1 week of dosing . Western blotting showed that DB05465 inhibited phosphorylation of P36888 in circulating leukemic blasts . Eight patients had P36888 -ITD mutations ; 5 of these were evaluable for assessment of DB05465 's antileukemic effect . Two of the 5 patients , treated at 525 mg and 700 mg twice daily , showed evidence of antileukemic activity , with decreases in both peripheral and bone marrow blasts . Tandutinib at the MTD ( 525 mg twice daily ) should be evaluated more extensively in patients with AML with P36888 -ITD mutations to better define its antileukemic activity . Q99527 Signaling in Spermatogenesis and Testicular Tumors . DB00286 play important roles in the regulation of testis development and spermatogenesis . Moreover , several evidences suggest that estrogen signaling can be involved in testicular tumorigenesis . The physiological effects of estrogen are mediated by the classical nuclear estrogen receptors P03372 and 2 , which regulate both genomic and rapid signaling events . In the recent years , a member of the seven-transmembrane G protein-coupled receptor family , Q99527 ( Q99527 ) , has been identified to promote estrogen action in target cells including testicular cells . Ours and other studies reported that Q99527 is expressed in normal germ cells ( spermatogonia , spermatocytes , spermatids ) , somatic cells ( Sertoli and Leydig cells ) , and it is also involved in mediating estrogen action during spermatogenesis and testis development . In addition , Q99527 seems to be involved in modulating estrogen-dependent testicular cancer cell growth . However , in this context , the effects of Q99527 stimulation on cell survival and proliferation appear to be cell type specific . This review summarizes the current knowledge on the functions regulated by estrogens and mediated by Q99527 in normal and tumor testicular cells . DB00659 : recent findings and future research directions . This article explores the mechanisms of action and the potential responder profile of acamprosate , a compound efficacious in relapse prevention of alcoholism . New evidence at the molecular and cellular level suggests that acamprosate attenuates hyper-glutamatergic states that occur during early abstinence and involves iono ( DB01221 ) - and metabotrotropic ( P41594 ) glutamate receptors along with augmented intracellular calcium release and electrophysiological changes . Thus mutant mice with enhanced glutamate levels exhibit higher alcohol consumption than wild type mice and respond better to acamprosate , demonstrating that acamprosate acts mainly on a hyper-glutamatergic system . This mode of action further suggests that acamprosate exhibits neuroprotective properties . In rats , cue-induced reinstatement behavior is significantly reduced by acamprosate treatment whereas cue-induced craving responses in alcohol-dependent patients seem not to be affected by this treatment . An ongoing study ( " Project Predict " ) defines specific responder profiles for an individualized use of acamprosate and naltrexone . Neurophysiological as well as psychometric data are used to define 2 groups of patients : " reward cravers " and " relief cravers " . While naltrexone should work better in the first group , acamprosate is hypothesized to be efficacious in the latter where withdrawal associated and/or cue induced hyper-glutamatergic states are thought to trigger relapse . Further research should target the definition of subgroups applying endophenotypic approaches , e.g. by detecting a hyperglutamatergic syndrome using MR spectroscopy . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . Protein kinase C-associated kinase regulates NF-κB activation through inducing IKK activation . Activation of the transcription factor NF-κB induced by extracellular stimuli requires IKKα and IKKβ kinase activity . How IKKα and IKKβ are activated by various upstream signaling molecules is not fully understood . We previously showed that protein kinase C-associated kinase ( P03952 , also known as P57078 /RIP4 ) , which belongs to the receptor-interacting protein ( RIP ) kinase family , mediates the B cell activating factor of the P01375 family ( Q9Y275 ) -induced NF-κB activation in diffuse large B cell lymphoma ( DLBCL ) cell lines . Here we have investigated the mechanism underlying NF-κB activation regulated by P03952 . Our results suggest that P03952 can activate both the classical and the alternative NF-κB activation pathways . P03952 associates with IKKα and IKKβ in mammalian cells and induces activation of both IKKα and IKKβ via phosphorylation of their serine residues 176/180 and 177/181 , respectively . Unlike other members of the RIP family that activate NF-κB through a kinase-independent pathway , P03952 appears to activate IKK and NF-κB mainly in a kinase-dependent manner . Suppression of P03952 expression by RNA interference inhibits phosphorylation of IKKα and IKKβ as well as activation of NF-κB in human cancer cell lines . Thus , P03952 regulates NF-κB activation by modulating activation of IKKα and IKKβ in mammalian cells . We propose that P03952 may provide a critical link between IKK activation and various upstream signaling cascades , and may represent a potential target for inhibiting abnormal NF-κB activation in human cancers . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . P35354 induction and prostaglandin E2 accumulation in squamous cell carcinoma as a consequence of epidermal growth factor receptor activation by imatinib mesylate . Imatinib mesylate is a novel anti-tumor agent useful in the clinical management of chronic myelogenous leukemia and gastrointestinal stromal tumors with minimal toxicity relative to other forms of cancer therapy . Its clinical activity and minimal toxicity are related to specific inhibition of cellular targets including P11274 - P00519 , platelet-derived growth factor receptor and c-kit kinases , resulting in the collapse of downstream signaling cascades important for transformation . In some patients , unexpected toxicities arise that are not associated with inhibition of any known cellular imatinib target . In this report , we investigated the effects of imatinib on squamous carcinoma cell signaling . Imatinib induced expression of P35354 in a dose-dependent manner with concomitant accumulation of prostaglandin E2 . P35354 induction by imatinib was initiated through epidermal growth factor ( P01133 ) receptor kinase activation and downstream signaling through mitogenic-activated protein kinase . P35354 induction by imatinib was blocked by Q02750 or P01133 receptor inhibition . Imatinib did not activate stressor cytokine-signaling pathways ( p38 kinase , nuclear factor-kB nuclear translocation ) or affect P23219 expression . Imatinib failed to activate P01133 receptor signals in other tumor types , suggesting that P35354 induction in imatinib-treated cells is mediated through release of autocrine factors expressed or activated in squamous tumors . P35354 induction by imatinib in squamous tumors derived from the head and neck region is unique with respect to other target-specific agents and may represent one of the unintended toxic effects of imatinib described in some patients . Human P20273 can not fully substitute murine P20273 functions in vivo , as shown in a new knockin mouse model . P20273 , an inhibitory co-receptor of the B-cell receptor , shows a B-cell-specific expression pattern and is expressed on most B-cell lymphomas . The anti- P20273 antibody DB04958 is in clinical trials for B-cell non-Hodgkin lymphoma and systemic lupus erythematosus , but shows a mostly unknown mode of action . We generated a new mouse model that expresses human P20273 instead of murine P20273 ( Huki P20273 mice ) , in which human P20273 can be targeted . Expression of human P20273 on the B cells of Huki P20273 mice does not generally interfere with B-cell development . However , Huki P20273 mice show a reduction of the population of mature recirculating B cells in the bone marrow and reduced transitional and marginal zone B cells in the spleen , phenotypes resembling that of P20273 -deficient mice . Similarly , enhanced P11274 -induced Ca(2+) signalling is observed in Huki P20273 mice , which also mount normal immune responses toward different classes of antigens . Huki P20273 B cells show a normal anti-hCD22 antibody-mediated endocytosis . In conclusion , human P20273 can not fully substitute for murine P20273 functions , possibly due to the changed intracellular tail of the protein or due to lower expression levels . Huki P20273 mice are a valuable new model for both antibody- and immunotoxin-mediated targeting of human P20273 . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling . Immunomodulatory influence of bone marrow-derived DB05914 on neuroinflammation in astrocyte cultures . The therapeutic benefits associated with DB05914 ( MSCs ) largely result from their immunomodulatory and neurotrophic properties . In this study , we evaluated the effects of MSCs on astrocyte cultures exposed to lipopolysaccharide . In response to this inflammatory trigger , astrocytes showed an increased expression of pro-inflammatory genes ( IL-1β , TNFα , P05231 ) , which was attenuated by pre-exposure to O60682 conditioned medium . Furthermore , mediators released by MSCs increased cell proliferation and altered the regulation of intermediate filaments ( P14136 , vimentin ) , pro-inflammatory enzymes ( P35228 , P35354 ) and receptors ( O00206 , P08571 , Q14832 , P41594 ) . These data demonstrate that MSCs influence diverse cell types participating in the response to neuroinflammation . Comparison of two polymer-based immunohistochemical detection systems : ENVISION+ and ImmPRESS . The non-specific background reaction produced in avidin-biotin-based immunohistochemistry , particularly after harsh antigen retrieval procedures , has promoted the use of non-avidin-biotin systems , yet there are few reports comparing the performance of non-avidin-biotin , polymer-based methods . In this study we compare two of these methods , ENVISION+trade mark and ImmPRESS , in animal tissues . We examined the immunoreactivity of 18 antigens in formalin-fixed , paraffin-embedded tissues . Antigens were located in the cytoplasmic membrane ( CD11d , P05107 and CD79a ) , cytoplasm ( calretinin , P23219 , P35354 , Glut-1 , HepPar 1 , P10721 , Melan A , tryptase and uroplakin III ) or nucleus ( Q2TAK8 , P09936 and thyroid transcription factor 1 ) . We also evaluated three infectious agents ( Aspergillus , calicivirus and West Nile virus ) . The staining with ENVISION+ or ImmPRESS was performed simultaneously for each antigen . The intensity of the reaction and background staining were scored . ImmPRESS yielded similar or higher reaction intensity than ENVISION+trade mark in 16/18 antigens . ImmPRESS produced abundant background with the other two antigens ( calretinin and P35354 ) , which hindered interpretation of the specific reaction . The cost of ImmPRESS was 25 % lower than for ENVISION+trade mark . Based on these results , ImmPRESS is a good polymer-based detection system for routine immunohistochemistry . Regulation of MicroRNA 183 by Cyclooxygenase 2 in Liver Is DEAD-Box Helicase p68 ( P17844 ) Dependent : Role in P01308 Signaling . Cyclooxygenase ( P36551 ) catalyzes the first step in prostanoid biosynthesis and exists as two isoforms . P23219 is a constitutive enzyme involved in physiological processes , whereas P35354 is induced by a variety of stimuli . MicroRNAs ( miRNAs ) are noncoding RNAs that function as key posttranscriptional regulators of gene expression . Although it is known that P35354 expression is regulated by miRNAs , there are no data regarding P35354 involvement in miRNA regulation . Considering our previous results showing that P35354 expression in hepatocytes protects against insulin resistance , we evaluated the role of P35354 in the regulation of a specific set of miRNAs implicated in insulin signaling in liver cells . Our results provide evidence of the molecular basis for a novel function of P35354 in miRNA processing . P35354 represses miRNA 23b ( miR-23b ) , miR-146b , and miR-183 expression in liver cells by increasing the level of DEAD-box helicase p68 ( P17844 ) through phosphatidylinositol 3-kinase ( PI3K ) /p300 signaling and by modulating the enzymatic function of the Drosha ( RNase type III ) complex through its physical association with P17844 . The decrease of miR-183 expression promotes protection against insulin resistance by increasing insulin receptor substrate 1 ( P35568 ) levels . These results indicate that the modulation of miRNA processing by P35354 is a key event in insulin signaling in liver and has potential clinical implications for the management of various hepatic dysfunctions . Cyclooxygenases in rat Leydig cells : effects of luteinizing hormone and aging . Previous studies suggested that increased Leydig cell cyclooxygenase ( P36551 )2 expression may be involved in the reduced testosterone production that characterizes aged Leydig cells . Our objective herein was to further elucidate the relationships among LH stimulation , Leydig cell P35354 and P23219 expression , aging , and testosterone production . Incubation of Leydig cells from young or aged rats with LH or dibutyryl DB02527 resulted in increases in both intracellular P35354 protein expression and testosterone production . P23219 expression did not respond to LH or dibutyryl DB02527 . Incubation of adult cells with a protein kinase A inhibitor suppressed the stimulatory effects of LH on P35354 and testosterone production . Short-term incubation of Leydig cells with TGF-alpha or IL-1beta also increased P35354 protein levels ; P05019 had no effect . In vivo , LH also was found to stimulate both P35354 and testosterone , but not P23219 . As reported previously , P35354 expression was greater in old than in young cells , and old Leydig cells responded to inhibition of P35354 in vitro with increased testosterone production . However , the effects of the P35354 inhibitors were not restricted to old cells ; young Leydig cells also responded to P35354 inhibition with increased testosterone production . This and the observation that the incubation of young or old cells with LH resulted in increased P35354 and testosterone production in both cases suggests that the relationship between P35354 and testosterone production is not unique to aged Leydig cells . Moreover , the close correlation between increases in P35354 and testosterone in LH-stimulated young and aged Leydig cells is difficult to reconcile with the contention that the increased expression of P35354 in aged cells is responsible for age-related suppression of Leydig cell testosterone production .

MH_train_1157

MH_train_1157

MH_train_1157

interacts_with DB04946?

multiple_choice

Q9NQ38 and P07550 haplotypes are risk factors for asthma in Mexican pediatric patients . BACKGROUND : Asthma is one of the most common respiratory diseases worldwide , and the complexity of its etiology has been widely documented . Chromosome 5q31-33 is one of the main loci implicated in asthma and asthma-related traits . P35225 , P08571 and P07550 , which are located in this risk locus , are among the genes most strongly associated with asthma susceptibility . OBJECTIVES : This study evaluated whether single-nucleotide polymorphisms or haplotypes at 5q31-33 conferred risk for asthma in Mexican-Mestizo pediatric patients . METHODS : We performed a case-controlled study including 851 individuals , 421 of them affected with childhood-onset asthma and 430 ethnically matched unaffected subjects . We used the TaqMan Allelic Discrimination Assay to genotype 20 single-nucleotide polymorphisms within P05113 , Q92878 , P35225 , P05112 , P08571 , Q9NQ38 , Q13639 , P07550 and P29460 . RESULTS : Although no association was detected for any risk allele , three Q9NQ38 haplotypes ( O75223 : p = 6 × 10(-6) ; AATC : p = 0.0001 ; AGTT : p = 0.0001 ) and five P07550 haplotypes ( AGGACC : p = 0.0014 ; AGGAAG : p = 0.0002 ; TGAGAG : p = 0.0001 ; AGGAAC : p = 0.0002 ; AAGGAG : p = 0.003 ) were associated with asthma . Notably , the AGTT Q9NQ38 haplotype exhibited a male gender-dependent association ( p = 7.6 × 10(-5) ) . CONCLUSION : Our results suggest that Q9NQ38 and P07550 haplotypes might play a role in the susceptibility to childhood-onset asthma . [ 5-hydroxytryptamine ( serotonin ) receptors -- nomenclature and classification of types and subtypes ] . 5-HT receptors represent a superfamily of receptors with the largest known number of receptor subtypes . At present 15 receptor subtypes of three groups has been recognized . The 5-HT1 subfamily of receptors contains subtypes P08908 , P28222 , P28221 , P28566 , and P30939 ; activation of all of them results in the inhibition of adenylylcyclase . The subfamily of 5-HT2 contains subtypes 5- Q13049 , P41595 , and P28335 ; their activation leads to the stimulation of P98160 . Finally , subfamily of miscellaneous 5-HT receptors contains subtypes 5- Q9H205 , Q13639 , 5-HT5 , P50406 , and P34969 ; some of them has been cloned , however , our knowledge on their function is still minimal . 5-HT receptors participate in many physiological functions and a disturbance in serotonergic neurotransmission might cause several types of disease . 5-HT plays an important role in depression ; to cure this disease , drugs which increase levels of this neurotransmitter are used . A new drug group called Selective Serotonin Reuptake Inhibitors ( SSRI ) has been recently discovered . These drugs block the reuptake of 5-HT into nerve endings . There is an intensive search for new selective agonists as well as antagonists which could be use not only in the classification of receptor subtypes but which also possess certain therapeutic potential . 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 . Blockade of Q07343 limits lung vascular permeability and lung inflammation in LPS-induced acute lung injury . Acute lung injury ( ALI ) , acute respiratory distress syndrome ( ARDS ) , is actually involved in an ongoing and uncontrolled inflammatory response in lung tissues . Although extensive studies suggested that phospodiesterase type 4B ( Q07343 ) may be related to inflammation , the underlying cell biological mechanism of ALI remains unclear . To further investigate the mechanism how Q07343 take part in inflammatory response and the maintenance of vascular integrity , we established the experimental model of ALI in vitro and in vivo . In vitro , we found that DB03849 , Diazepam and Q07343 knockout could potently inhibit the LPS-induced NF-κB activation and inflammatory response in multiple cell types , including lung epithelial cells ( A549 ) , pulmonary microvascular endothelial cells ( PMVECs ) and vascular smooth muscle cells ( VSMCs ) . Besides , Q07343 deletion attenuated the LPS-induced ROS generation . In vivo , Q07343 deletion could attenuate the lung water content , histological signs of pulmonary injury and elevate the ratio of partial pressure of arterial O2 to fraction of inspired O2 ( PaO2/FIO2 ratio ) . Additionally , Q07343 deletion reduced LPS-induced vascular permeability . Collectively , our results strongly indicates that Q07343 is a valid target for anti-ALI . Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 [ ( R ) -(-)-2-(4-bromo-2-fluorobenzyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-4-spiro-3'-pyrrolidine-1,2',3,5'-tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K(i) = 10(-)(10) M ) 2000-fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R- and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP(+) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP(+) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP(+) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and/or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4-bromo-2-fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) . The heteromeric GABA-B receptor recognizes G-protein alpha subunit C-termini . The recently cloned GABA-B receptors are related to the metabotropic glutamate receptors ( mGlu receptors ) , the Ca2+-sensing receptor and one group of vomeronasal receptors . The GABA-B receptors likely function in a heterodimeric form , constituted of Q9UBS5 and O75899 . This novel feature in the G-protein coupled receptors ( GPCRs ) structure raises questions as to the mechanism of recognition of G-proteins by such receptors . In the present study we show that the Q9UBS5 and BR2 subunits form a functional receptor that recognizes the extreme C-termini of the G alpha i and G alpha o proteins when expressed in HEK293 cells . Indeed , heteromeric Q9UBS5 /BR2 receptors do not activate P98160 when co-expressed with G alpha q , but do so when co-expressed with the chimeric G alpha qi5 or G alpha qo5 subunits , the G alpha q subunit in which the 5 C-terminal residues are those of G alpha i or G alpha o , respectively . Interestingly , the heteromeric GABA-B receptor did not activate the chimeric G alpha qz5 subunit that contains the 5 C-terminal residues of G alpha z . Among the three residues that are distinct between G alpha qo5 and G alpha qz5 ( at position -5 , -4 and -1 ) , the amino acid residue at position -4 of G alpha o proteins is critical for specifying the coupling selectivity with the receptor and residue -5 influences the coupling efficacy . Interestingly , these findings correspond to data obtained with the Q14416 receptor , a distant relative of GABA-B proteins . This shows that the same molecular determinants of the G-protein alpha-subunits are involved in the specific recognition of both the heteromeric GABA-B receptors and the other GPCRs . ZNF804a regulates expression of the schizophrenia-associated genes Q9NQE7 , P21964 , Q07343 , and P14416 . ZNF804a was identified by a genome-wide association study ( GWAS ) in which a single nucleotide polymorphism ( SNP rs1344706 ) in ZNF804a reached genome-wide statistical significance for association with a combined diagnosis of schizophrenia ( SZ ) and bipolar disorder . Although the molecular function of ZNF804a is unknown , the amino acid sequence is predicted to contain a C2H2-type zinc-finger domain and suggests ZNF804a plays a role in DNA binding and transcription . Here , we confirm that ZNF804a directly contributes to transcriptional control by regulating the expression of several SZ associated genes and directly interacts with chromatin proximal to the promoter regions of Q9NQE7 and P21964 , the two genes we find upregulated by ZNF804a . Using immunochemistry we establish that ZNF804a is localized to the nucleus of rat neural progenitor cells in culture and in vivo . We demonstrate that expression of ZNF804a results in a significant increase in transcript levels of Q9NQE7 and P21964 , relative to GFP transfected controls , and a statistically significant decrease in transcript levels of Q07343 and P14416 . Furthermore , we show using chromatin immunoprecipitation assays ( ChIP ) that both epitope-tagged and endogenous ZNF804a directly interacts with the promoter regions of Q9NQE7 and P21964 , suggesting a direct upregulation of transcription by ZNF804a on the expression of these genes . These results are the first to confirm that ZNF804a regulates transcription levels of four SZ associated genes , and binds to chromatin proximal to promoters of two SZ genes . These results suggest a model where ZNF804a may modulate a transcriptional network of SZ associated genes . Method for isolating tight-binding inhibitors of rat lens aldose reductase . Numerous animal studies indicate that aldose reductase inhibitors ( ARIs ) are beneficial for the prevention or amelioration of diabetic complications such as neuropathy , nephropathy and the ocular complications of cataract , retinopathy and keratopathy . To aid in the identification of novel potent ARIs , we have previously developed a screening method that is based on the formation of a non-covalent ternary tight-binding enzyme-inhibitor-nucleotide ( AR- Q9Y4X5 -NADPH ) complex that can be isolated using YM-10 filter units . Here , we report a modification of this method that permits us to rapidly identify tight binding ARIs that are isolated by denaturation from AR- Q9Y4X5 -NADPH complexes that are free of possible contamination resulting from the reaction of methanol with the YM-10 filter units . For the development of this procedure , nine structurally diverse ARIs were mixed with purified recombinant rat lens aldose reductase ( RLAR ) bound with NADPH to form tight-binding RLAR- Q9Y4X5 -NADPH complexes . These complexes were purified by high pressure Sephadex 75 size exclusion chromatography using ammonium acetate buffer and the formation of each complex was confirmed by electrospray ionisation mass spectrometry ( P19957 -MS ) . Each of the complexes was then denatured with methanol , rechromatographed on the size exclusion column , and the identity of the bound ARIs was confirmed by P19957 -MS . The apparent Q9Y4X5 binding with aldose reductase to form a tight binding Q9Y4X5 complex appeared proportional to their IC50 values . This procedure allows for the rapid identification of tight binding ARIs with apparent IC50s < 0.1 microm . The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . 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 . Molecular and behavioral effects mediated by Gs-coupled adenosine A2a , but not serotonin 5-Ht4 or 5-Ht6 receptors following antipsychotic administration . Typical antipsychotic agents are potent antagonists of Gi-coupled dopamine D2 receptors , but their mechanisms of action following this initial blockade remain poorly understood . We hypothesized that in striatal neurons , interruption of this inhibitory dopamine D2 input would unmask endogenous striatal Gs-coupled receptors . An increase in DB02527 levels generated by these unopposed receptors would then lead to the well-described behavioral and molecular effects of antipsychotic administration such as catalepsy and striatal c-fos and neurotensin gene transcription . We examined three striatal Gs-coupled receptor systems ( serotonin Q13639 , serotonin P50406 and adenosine A2a ) to assess their potential involvement in the mechanism of action of the typical antipsychotic haloperidol . Antagonists of each of these three receptor systems together with a 1 mg/kg dose of haloperidol were co-administered to Sprague-Dawley rats , and both the degree of catalepsy produced in the animals and the induction of striatal c-fos and neurotensin messenger RNAs were measured . Both the specific adenosine A2a antagonist 8-(3-chlorostyryl)-caffeine and the general adenosine antagonist theophylline reduced haloperidol-dependent induction of striatal neurotensin and c-fos messenger RNA . Administration of these agents also greatly reduced the degree of catalepsy induced by haloperidol . Antagonists of the P50406 receptor failed to block the induction of striatal messenger RNAs , but the P50406 antagonist clozapine ( an important atypical antipsychotic agent in its own right ) was a potent inhibitor of catalepsy . Q13639 agents were unable to alter haloperidol 's effects on striatal messenger RNA levels or catalepsy . We conclude that the striatal Gs-coupled adenosine A2a receptor is an important mediator of the molecular and behavioral sequelae following haloperidol administration . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients . DB00147 phosphate-responsive seizures in a patient with cerebral folate deficiency ( P00746 ) and congenital deafness with labyrinthine aplasia , microtia and microdontia ( P24043 ) . We present an 8-year-old boy with folate receptor alpha ( FRα ) defect and congenital deafness with labyrinthine aplasia , microtia and microdontia ( P24043 syndrome ) . Both conditions are exceptionally rare autosomal recessive inherited diseases mapped to 11q13 . Our patient was found to have novel homozygous nonsense mutations in the P15328 gene ( p.R204X ) , and P11487 gene ( p.C50X ) . While the FRα defect is a disorder of brain-specific folate transport accompanied with cerebral folate deficiency ( P00746 ) causing progressive neurological symptoms , P24043 syndrome is a solely malformative condition , with normal physical growth and cognitive development . Our patient presented with congenital deafness , hypotonia , dysphygia and ataxia in early childhood . At the age of 6 years he developed intractable epilepsy , and deteriorated clinically with respiratory arrest and severe hypercapnea at the age of 8 years . In contrast to the previously published patients with a P15328 gene defect , our patient presented with an abnormal l-dopa metabolism in P04141 and high 3-O-methyl-dopa . Upon oral treatment with folinic acid the boy regained consciousness while the epilepsy could be successfully managed only with additional pyridoxal 5'-phosphate ( PLP ) . This report pinpoints the importance of P04141 folate investigations in children with unexplained progressive neurological presentations , even if a malformative syndrome is obviously present , and suggests a trial with PLP in folinic acid-unresponsive seizures . 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 . 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 . 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 . Isolation and structural characterization of the photolysis products of etoricoxib . DB01628 is a potent and novel selective inhibitor of cyclooxygenase-2 ( P35354 ) which has been developed for the treatment of osteoarthritis , rheumatoid arthritis and several other inflammatory conditions . To support clinical pharmacokinetics studies , a method for the determination of etoricoxib in human plasma was developed . During the development of the method it was found that highly fluorescent products were formed when etoricoxib was exposed to UV light ( 254 nm ) . The formation of highly fluorescent products was the basis for the development of a highly sensitive HPLC/fluorescent assay for the indirect determination of etoricoxib in human plasma ; the limit of quantification ( LOQ ) was 1 ng/mL . To unequivocally determine the chemical structures of the photolysis products of etoricoxib , a series of studies was conducted . When etoricoxib was irradiated online in a photochemical reactor , three products were detected in an HPLC-UV system . These products were characterized by HPLC-UV-fluorescence and HPLC-MS/MS . Possible structures of these products were proposed based on these data . The major photolysis products of etoricoxib were further isolated and their structures were elucidated using NMR and HPLC-NMR . The results of these experiments indicate that etoricoxib undergoes a photocyclization reaction when irradiated with UV light ( 254 nm ) , leading to the formation of two major isomeric photocyclization products . DB05229 sodium , prostacyclin analogue , attenuates glomerular hyperfiltration and glomerular macrophage infiltration by modulating ecNOS expression in diabetic rats . Stable prostacyclin analogue , beraprost sodium ( BPS ) has recently been reported to attenuate glomerular hyperfiltration in diabetic rats , however , the mechanism has been still unknown . We previously reported that overexpression of endothelial cell nitric oxide synthase ( ecNOS ) in afferent arterioles and glomeruli induce inappropriate dilatation of afferent arterioles and glomerular hyperfiltration through overproduction of nitric oxide in early stage of diabetic nephropathy . In this study , we tested the hypothesis that BPS ameliorates glomerular hyperfiltration through modulating ecNOS expression in diabetic nephropathy . Furthermore , we examined the effects of BPS on the expression of intercellular adhesion molecule-1 ( P05362 ) and macrophage infiltration in diabetic glomeruli , because glomerular hyperfiltration induces the expression of P05362 resulting in macrophage infiltration . Male Sprague-Dawley ( SD ) rats were administered continuously with BPS for 4 weeks after induction of diabetes by streptozotocin . In diabetic rats , the diameters of afferent arterioles , glomerular volume , creatinine clearance and urinary excretion of albumin and NO2/NO3 were increased as compared with non-diabetic control rats . Treatment with BPS improved these changes . The expression of ecNOS was increased in afferent arterioles and glomeruli in diabetic rats and suppressed by BPS . P43119 was expressed along afferent arterioles . Our results suggest that BPS attenuates glomerular hyperfiltration by modulating ecNOS expression in early stage of diabetic nephropathy . Moreover , BPS may inhibit P05362 -dependent infiltration of macrophages in diabetic glomeruli . 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 . P43119 up-regulates the expression of angiogenic genes in human endometrium via cross talk with epidermal growth factor Receptor and the extracellular signaling receptor kinase 1/2 pathway . DB01240 ( P06744 ) is a member of the prostanoid family of lipid mediators that mediates its effects through a seven-transmembrane G protein-coupled receptor ( IP receptor ) . Recent studies have ascertained a role for prostanoid-receptor signaling in angiogenesis . In this study we examined the temporal-spatial expression of the IP receptor within normal human endometrium and additionally explored the signaling pathways mediating the role of IP receptor in activation of target angiogenic genes . Quantitative RT-PCR analysis demonstrated the highest endometrial expression of the IP receptor during the menstrual phase compared with all other stages of the menstrual cycle . Immunohistochemical analysis localized the site of IP receptor expression to the glandular epithelial compartment with stromal and perivascular cell immunoreactivity . Expression of the immunoreactive IP receptor protein was greatest during the proliferative and early secretory phases of the menstrual cycle . To explore the role of the IP receptor in glandular epithelial cells , we used the Ishikawa endometrial epithelial cell line . Stimulation of Ishikawa cells and human endometrial biopsy explants with 100 nm iloprost ( a P06744 analog ) rapidly activated P27361 /2 signaling and induced the expression of proangiogenic genes , basic fibroblast growth factor , angiopoietin-1 , and angiopoietin-2 , in an epidermal growth factor receptor ( P00533 ) -dependent manner . Furthermore , P00533 colocalized with IP receptor in the glandular epithelial compartment . These data suggest that P06744 -IP interaction within glandular epithelial cells can promote the expression of proangiogenic genes in human endometrium via cross talk with the P00533 . DB05595 in lung cancer . DB00158 is essential for proliferating cells and folate transport pathways and folate-dependent metabolic processes show promise as targets for anti-neoplastic therapy . DB00158 receptor α ( P15328 ) , a folate transporter , is an attractive target for anti-neoplastic therapy due to its high affinity for folate , restricted range of expression in normal tissue and differential over-expression in malignant tissue . P15328 is expressed in non-small cell lung cancer , with a higher expression in adenocarcinoma compared with squamous cell carcinoma . DB05595 is a monoclonal antibody targeting P15328 which in pre-clinical studies led to cytotoxicity of P15328 -expressing cells , inhibited tumor growth in animal models and showed limited reactivity with normal tissue . In phase I/II trials , farletuzumab was well tolerated as a single-agent and in combination , without additive toxicity with chemotherapy . An ongoing phase II , double blind , placebo-controlled study is evaluating farletuzumab in patients with P15328 expressing metastatic adenocarcinoma of lung . 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 . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS )

MH_train_1158

MH_train_1158

MH_train_1158

interacts_with DB00622?

multiple_choice

High-resolution X-ray structure of isoaspartyl dipeptidase from Escherichia coli . Q7L266 from Escherichia coli functions in protein degradation by catalyzing the hydrolysis of beta-L-isoaspartyl linkages in dipeptides . The best substrate for the enzyme reported thus far is iso- DB00128 - DB00149 . Here we report the X-ray analysis of the enzyme in its resting state and complexed with aspartate to 1.65 and 2.1 A resolution , respectively . The quaternary structure of the enzyme is octameric and can be aptly described as a tetramer of dimers . Each subunit folds into two distinct domains : the N-terminal region containing eight strands of mixed beta-sheet and the C-terminal motif that is dominated by a ( beta,alpha ) (8)-barrel . A binuclear zinc center is located in each subunit at the C-terminal end of the ( beta,alpha ) (8)-barrel . Ligands to the binuclear metal center include DB00117 68 , DB00117 70 , DB00117 201 , DB00117 230 , and DB00128 285 . The two zincs are bridged by a carboxylated lysine residue ( Lys 162 ) and a solvent molecule , most likely a hydroxide ion . The product of the reaction , aspartate , binds to the enzyme by displacing the bridging solvent with its side chain functional group . From this investigation it is proposed that the reaction mechanism of the enzyme proceeds through a tetrahedral intermediate and that the bridging solvent attacks the re face of the carbonyl carbon of the scissile peptide bond . This structural analysis confirms the placement of isoaspartyl dipeptidase into the urease-related amidohydrolase superfamily . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . 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 . P62158 regulates Ca2+-sensing receptor-mediated Ca2+ signaling and its cell surface expression . The Ca(2+)-sensing receptor ( P41180 ) is a member of family C of the GPCRs responsible for sensing extracellular Ca(2+) ( [Ca(2+)](o) ) levels , maintaining extracellular Ca(2+) homeostasis , and transducing Ca(2+) signaling from the extracellular milieu to the intracellular environment . In the present study , we have demonstrated a Ca(2+)-dependent , stoichiometric interaction between P62158 and a P62158 -binding domain ( CaMBD ) located within the C terminus of P41180 ( residues 871-898 ) . Our studies suggest a wrapping around 1-14-like mode of interaction that involves global conformational changes in both lobes of P62158 with concomitant formation of a helical structure in the CaMBD . More importantly , the Ca(2+)-dependent association between P62158 and the C terminus of P41180 is critical for maintaining proper responsiveness of intracellular Ca(2+) responses to changes in extracellular Ca(2+) and regulating cell surface expression of the receptor . Gene expression studies provide clues to the pathogenesis of uterine leiomyoma : new evidence and a systematic review . BACKGROUND : Uterine leiomyomas are extremely common and a major cause of pelvic pain , bleeding , infertility , and the leading indication for hysterectomy . Familial and epidemiological studies provide compelling evidence that genetic alterations play an important role in leiomyoma development . METHODS : Using Affymetrix U133A GeneChip we analysed expression profiles of 22,283 genes in paired samples of leiomyoma and adjacent normal myometrium . We compared our results with previously published data on gene expression in uterine leiomyoma and identified the overlapping gene alterations . RESULTS : We detected 80 genes with average differences of > or = 2-fold and false discovery rates of < 5 % ( 14 overexpressed and 66 underexpressed ) . A comparative analysis including eight previous gene expression studies revealed eight prominent genes ( P07327 , P18847 , P29373 , O00622 , Q07507 , P42262 , P01344 , Q5EB52 ) identified by at least five different studies , eleven genes ( P00352 , P25063 , P29279 , O43602 , P28562 , P01100 , O60829 , P24592 , P41222 , P43115 , P04818 ) reported by four studies , twelve genes ( ABCA , P04083 , Q15847 , O00585 , P38936 , Q14194 , P54849 , P03372 , FY , Q99683 , P37173 , P35625 ) identified by three studies , and 40 genes reported by two different studies . CONCLUSIONS : Review of gene expression data revealed concordant changes in genes regulating retinoid synthesis , IGF metabolism , TGF-beta signaling and extracellular matrix formation . Gene expression studies provide clues to the relevant pathways of leiomyoma development . 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 . 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- 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 . Quantification and modeling of tripartite P06729 - , CD58FC chimera (alefacept)- , and CD16-mediated cell adhesion . DB00092 is a chimeric protein combining P19256 immunoglobulin-like domain 1 with human IgG1 Fc . DB00092 mediates adhesion by bridging P06729 on T cells to activating Fc receptors on effector cells , but the equilibrium binding parameters have not been determined . DB00092 mediated T cell killing by NK cells and adhesion between P06729 - and CD16-expressing cells at an optimum concentration of 100 nM . We introduce novel measurements with supported planer bilayers , from which key two-dimensional and three-dimensional parameters can be determined by data fitting . DB00092 competitively inhibited cell bilayer adhesion mediated by the P06729 - P19256 interaction . DB00092 mediated maximal adhesion of P06729 (+) T cells to O75015 , an Fc receptor , in planar bilayers at 500 nM . A mechanistic model for alefacept-mediated cell-bilayer adhesion allowed fitting of the data and determination of two-dimensional binding parameters . These included the density of bonds in the adhesion area , which grew to maintain a consistent average bond density of 200 molecules/microm(2) and two-dimensional association constants of 3.1 and 630 microm(2) for bivalently and monovalently bound forms of alefacept , respectively . The maximum number of CD16 bound and the fit value of 4,350 P06729 per cell are much lower than the 40,000 P06729 per cell measured with anti- P06729 Fab . These results suggest that additional information is needed to correctly predict DB00092 -mediated bridge formation . EP3 receptor isoforms are differentially expressed in subpopulations of primate granulosa cells and couple to unique G-proteins . DB00917 ( DB00917 ) produced within the ovarian follicle is necessary for ovulation . DB00917 is recognized by four distinct G-protein-coupled receptors . Among them , P43115 ( also known as EP3 ) is unique in that mRNA splicing generates multiple isoforms . Each isoform has a distinct amino acid composition in the C-terminal region , which is involved in G-protein coupling . To determine whether monkey EP3 isoforms couple to different G-proteins , each EP3 isoform was expressed in Chinese hamster ovary cells , and intracellular signals were examined after stimulation with the EP3 agonist sulprostone . Stimulation of EP3 isoform 5 ( EP3-5 ) reduced DB02527 in a pertussis toxin ( PTX ) -sensitive manner , indicating involvement of Gαi . Stimulation of EP3-9 increased DB02527 , which was reduced by the general G-protein inhibitor GDP-β-S , and also increased intracellular calcium , which was reduced by PTX and GDP-β-S . So , EP3-9 likely couples to both Gαs and a PTX-sensitive G-protein to regulate intracellular signals . Stimulation of EP3-14 increased DB02527 , which was further increased by PTX , so EP3-14 likely regulates DB02527 via multiple G-proteins . Granulosa cell expression of all EP3 isoforms increased in response to an ovulatory dose of human chorionic gonadotropin . Two EP3 isoforms were differentially expressed in functional subpopulations of granulosa cells . EP3-5 was low in granulosa cells at the follicle apex while EP3-9 was high in cumulus granulosa cells . Differential expression of EP3 isoforms may yield different intracellular responses to DB00917 in granulosa cell subpopulations , contributing to the different roles played by granulosa cell subpopulations in the process of ovulation . A conserved role for Drosophila Neuroglian and human Q9NUQ9 - P62158 in central-synapse formation . BACKGROUND : Drosophila Neuroglian ( Nrg ) and its vertebrate homolog Q9NUQ9 - P62158 are cell-adhesion molecules ( P62158 ) that have been well studied in early developmental processes . Mutations in the human gene result in a broad spectrum of phenotypes ( the Q7L266 -syndrome ) that include devastating neurological disorders such as spasticity and mental retardation . Although the role of Q9NUQ9 -CAMs in neurite extension and axon pathfinding has been extensively studied , much less is known about their role in synapse formation . RESULTS : We found that a single extracellular missense mutation in nrg(849) mutants disrupted the physiological function of a central synapse in Drosophila . The identified giant neuron in nrg(849) mutants made a synaptic terminal on the appropriate target , but ultrastructural analysis revealed in the synaptic terminal a dramatic microtubule reduction , which was likely to be the cause for disrupted active zones . Our results reveal that tyrosine phosphorylation of the intracellular ankyrin binding motif was reduced in mutants , and cell-autonomous rescue experiments demonstrated the indispensability of this tyrosine in giant-synapse formation . We also show that this function in giant-synapse formation was conserved in human Q9NUQ9 - P62158 but neither in human Q9NUQ9 - P62158 with a pathological missense mutation nor in two isoforms of the paralogs NrCAM and O94856 . CONCLUSIONS : We conclude that Nrg has a function in synapse formation by organizing microtubules in the synaptic terminal . This novel synaptic function is conserved in human Q9NUQ9 - P62158 but is not common to all Q9NUQ9 -type proteins . Finally , our findings suggest that some aspects of Q9NUQ9 - P62158 -related neurological disorders in humans may result from a disruption in synapse formation rather than in axon pathfinding . Involvement of multiple protein kinases in CD3-mediated activation of human T lymphocytes . The role of different protein kinases in the process of T cell activation has been studied using several inhibitors . The model we adopted was the activation of PBMC by monoclonal antibody OKT3 . The results obtained confirm that PKC and PTK are involved . Thus , the inhibitors H-7 , staurosporine , and genistein exerted a dose-dependent inhibition of P06729 up-regulation , CD25 expression , P60568 production , and cellular proliferation . On the other hand , our data indicate that PKA is not involved since the inhibitor HA1004 was ineffective . W-7 , an inhibitor of Ca(2+)- P62158 protein kinases , inhibited OKT3-induced modulation of cell-surface markers and PBMC proliferation , whereas a slight increase in P60568 release was detected at the highest dose used ( 20 microM ) . Using the MLCK inhibitor ML-9 , we extended our studies to the myosin light chain kinase , which influences the organization of the cytoskeleton . ML-9-inhibited PBMC activation in terms of modulation of cell-surface markers and proliferation but stimulated P60568 production . Similar results were obtained using the cytoskeleton disruptors demecolcine and cytochalasin B . Taken together the data described herein indicate that T cell activation is a complex event in which , aside from classical signal transduction-associated kinases PKC and PTK , at least two other kinases , Ca(2+)- P62158 kinases and MLCK , seem to be involved , the latter probably through correct assembly of the cytoskeleton . 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 . A Lactobacillus rhamnosus strain induces a heme oxygenase dependent increase in Foxp3+ regulatory T cells . We investigated the consequences of feeding with a Lactobacillus species on the immune environment in P07902 , and the role of dendritic cells and heme oxygenase-1 in mediating these responses . Feeding with a specific strain of Lactobacillus rhamnosus induced a significant increase in P01730 +CD25+Foxp3+ functional regulatory T cells in P07902 . This increase was greatest in the mesenteric lymph nodes and associated with a marked decrease in P01375 and IFNγ production . Dendritic cell regulatory function and P09601 expression was also increased . The increase in Foxp3+ T cells could be prevented by treatment with a heme oxygenase inhibitor . However , neither inhibition of heme oxygenase nor blockade of P22301 and TGFβ prevented the inhibition of inflammatory cytokine production . In conclusion Lactobacillus feeding induced a tolerogenic environment in P07902 . P09601 was critical to the enhancement of Foxp3+ regulatory T cells while additional , as yet unknown , pathways were involved in the down-regulation of inflammatory cytokine production by T cells . [ Effectiveness of the screening programme for galactosemia. New strategy in Poland ] . Galactosemia is an autosomal recessive disease related to deficiency of one of three different enzymes involved in the metabolism of galactose : galactokinase ( P51570 ) , galactoso-J-phosphate uridyltransferase ( P07902 ) or DB03501 -4-epimerase ( Q14376 ) . Classic galactosemia is due to P07902 deficiency and is the most common . Longitudinal studies have shown that in spite of early diagnosis and early treatment of children with galactosemia detected in the mass screening programme , the results are poor and mental retardation as well as other complications are of similar severity as in children diagnosed clinically without screening . In many investigations it was also proved that some impairments developed already in the prenatal period . Therefore , many countries among them also Poland , stopped mass screening for galactosemia . At present , in Poland the procedure strategy in galactosemic children and their families include : diagnosis of new cases on the basis of clinical symptoms , selective screening in high-risk families , prophylactic lactose-free diet for mothers during pregnancy . Such management can help to prevent clinical manifestations in newborns and prevent death in the early period of life . P13688 impedes thyroid cancer growth but promotes invasiveness : a putative mechanism for early metastases . P13688 , also known as biliary glycoprotein ( BGP ) , CD66a , Q00839 and C- P62158 , is a member of the P06731 immunoglobulin superfamily . P13688 is a putative tumor suppressor based on diminished expression in some solid neoplasms such as colorectal carcinoma . However , P13688 is overexpressed in some tumors such as non-small cell lung cancer . To clarify the mechanism of action of this cell adhesion molecule , we studied thyroid carcinoma that has a spectrum of morphologies and variable behavior allowing separation of proliferation from invasion and metastasis . P13688 is expressed in thyroid carcinoma cell lines derived from tumors that exhibit aggressive behavior . Introduction of P13688 into endogenously deficient WRO cells resulted in reduced cell cycle progression associated with P38936 upregulation and diminished Rb phosphorylation . Forced P13688 expression enhanced cell-matrix adhesion and migration and promoted tumor invasiveness . Conversely , small interfering RNA ( siRNA ) -mediated downregulation of P13688 expression in Q9BYG7 cells accelerated cell cycle progression and significantly enhanced tumor size in xenografted mice . P13688 is not appreciably expressed in normal thyroid tissue or benign thyroid tumors . In a human thyroid tissue array , P13688 reactivity was associated with metastatic spread but not with increased tumor size . These findings identify P13688 as a unique mediator that restricts tumor growth whereas increasing metastatic potential . Our data highlight a complex repertoire of actions providing a putative mechanism underlying the spectrum of biologic behaviors associated with thyroid cancer . 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 . Drugs targeting nitric oxide synthase for migraine treatment . INTRODUCTION : Ample evidence that nitric oxide ( NO ) is a causative molecule in migraine has encouraged research to develop drugs that target the NO-cGMP cascade for migraine treatment . NO synthase ( NOS ) inhibition is an innovative therapeutic principle . AREAS COVERED : This paper reviews the rationale underlying NOS inhibition in migraine treatment . It also provides a review on the efficacy and safety data for NOS inhibitors ( nonselective NOS inhibitor L-N(G)-methyl-arginine hydrochloride [ L-NMMA ] , selective inducible NOS [ P35228 ] inhibitors GW273629 and GW274150 , combined neuronal NOS [ P29475 ] inhibitor and P28222 /1D receptor agonist DB06096 ) in acute or preventive migraine treatment . EXPERT OPINION : The data highlighted herein , from four placebo-controlled trials and 1 open-labeled clinical trial using 4 different NOS inhibitors on a total of 705 patients , provide convincing efficacy data only for the nonselective NOS inhibitor L-NMMA . Unfortunately , this NOS inhibitor raises cardiovascular safety concerns and has an unfavorable pharmacokinetic profile . As experimental studies predicted , P35228 inhibitors are ineffective in migraine . Still , upcoming selective P29475 inhibitors are a hope for migraine treatment , with the P29475 isoform being most clearly involved in trigeminovascular transmission and central sensitization . Future studies should help to clarify whether NOS inhibition is equally fruitful in acute and preventive treatment . It should also clarify if P29475 inhibition holds promise as a therapeutic tool for the treatment of chronic migraine and other forms of headache . DB00399 -induced IPP/ApppI production in vivo . Bisphosphonates are currently the most important class of anti-resorptive drugs used for the treatment of diseases involving excess bone resorption . Recently we discovered a new mechanism of action for bisphosphonates . Previously it has been shown that nitrogen-containing bisphosphonates ( N-BPs ) are not metabolized . However , our studies revealed that N-BPs induce formation of a novel pro-apoptotic DB00171 analog ( ApppI ) , as a consequence of the inhibition of P14324 in the mevalonate pathway , and the subsequent accumulation of isopentenyl pyrophosphate ( IPP ) in vitro . The primary aim of the current study was to determine whether zoledronic acid ( a N-BP ) induces IPP/ApppI formation in vivo . Mass spectrometry was used to identify whether in vivo administration of zoledronic acid-induced IPP/ApppI production by mouse peritoneal macrophages or bone marrow cells . IPP/ApppI could be detected in extracts from peritoneal macrophages isolated from zoledronic acid-treated animals . Increasing IPP/ApppI accumulation was determined up to 7 days after drug injection , indicating prolonged P14324 inhibition by zoledronic acid . Importantly , this is the first report of in vivo production of ApppI , supporting the biological significance of this molecule . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . Expression of adhesion molecules and c-kit on P28906 + hematopoietic progenitor cells : comparison of cytokine-mobilized blood stem cells with normal bone marrow and peripheral blood . We assessed the expression of the adhesion molecules leukocyte function antigen-1 ( LFA-1 , CD11a ) , intercellular adhesion molecule-1 ( P05362 , CD54 ) , homing-associated cell adhesion molecule ( H- P62158 , P16070 ) , and c-kit ( stem cell factor receptor ) on the P28906 + progenitor population from the leukapheresis products of 23 patients ( LP P28906 + ) . For blood stem cell collection granulocyte colony-stimulating factor ( DB00099 ) or interleukin-3/granulocyte-macrophage colony-stimulating factor ( P08700 /GM- P04141 ) was administered after cytotoxic chemotherapy . Furthermore , bone marrow- and blood-derived P28906 + progenitor cells from 6 normal volunteers ( BM and PB P28906 + ) were analyzed . LFA-1 expression was higher on PB P28906 + ( 88.2 +/- 2.5 % , mean +/- SEM ) than on BM P28906 + ( 75.3 +/- 4.3 % ) . Following cytokine administration , LFA-1 was expressed on only 59.7 +/- 3.7 % of LP P28906 + at a low fluorescence intensity , suggesting that down-regulation of LFA-1 may facilitate the egress of cells from the bone marrow and prolong their circulation . In contrast , P05362 was weakly positive on P28906 + cells from all sources . P16070 was expressed on the vast majority of P28906 + cells ( > 95 % ) in all samples studied . The highest proportion of P28906 + cells costaining for c-kit was found in normal bone marrow ( 32.2 +/- 3.3 % ) . In normal peripheral blood and after cytokine mobilization , fewer of the P28906 + cells weakly expressed c-kit ( < 15 % ) . The low percentage and level of c-kit expression may indicate that the majority of cytokine-mobilized P28906 + cells are lineage-committed progenitor cells , as reflected by the coexpression pattern for P28907 , HLA-DR , and P20138 .

MH_train_1159

MH_train_1159

MH_train_1159

interacts_with DB00338?

multiple_choice

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 . Novel phenolic antioxidants as multifunctional inhibitors of inducible P19320 expression for use in atherosclerosis . A series of novel phenolic compounds has been discovered as potent inhibitors of P01375 -inducible expression of vascular cell adhesion molecule-1 ( P19320 ) with concurrent antioxidant and lipid-modulating properties . Optimization of these multifunctional agents led to the identification of 3a ( DB05399 ) as a clinical candidate with demonstrated efficacies in animal models of atherosclerosis and hyperlipidemia . Enhanced killing of cancer cells by poly(ADP-ribose) polymerase inhibitors and topoisomerase I inhibitors reflects poisoning of both enzymes . Poly(ADP-ribose) polymerase-1 ( P09874 ) plays critical roles in the regulation of DNA repair . Accordingly , small molecule inhibitors of PARP are being developed as agents that could modulate the activity of genotoxic chemotherapy , such as topoisomerase I poisons . In this study we evaluated the ability of the PARP inhibitor veliparib to enhance the cytotoxicity of the topoisomerase I poisons topotecan and camptothecin ( CPT ) . DB07232 increased the cell cycle and cytotoxic effects of topotecan in multiple cell line models . Importantly , this sensitization occurred at veliparib concentrations far below those required to substantially inhibit poly(ADP-ribose) polymer synthesis and at least an order of magnitude lower than those involved in selective killing of homologous recombination-deficient cells . Further studies demonstrated that veliparib enhanced the effects of CPT in wild-type mouse embryonic fibroblasts ( MEFs ) but not Parp1(-/-) MEFs , confirming that P09874 is the critical target for this sensitization . Importantly , parental and Parp1(-/-) MEFs had indistinguishable CPT sensitivities , ruling out models in which P09874 catalytic activity plays a role in protecting cells from topoisomerase I poisons . To the contrary , cells were sensitized to CPT in a veliparib-independent manner upon transfection with P09874 E988K , which lacks catalytic activity , or the isolated P09874 DNA binding domain . These results are consistent with a model in which small molecule inhibitors convert P09874 into a protein that potentiates the effects of topoisomerase I poisons by binding to damaged DNA and preventing its normal repair . DB04849 : a highly potent , orally bioavailable , vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor for the treatment of cancer . Inhibition of vascular endothelial growth factor-A ( P15692 ) signaling is a promising therapeutic approach that aims to stabilize the progression of solid malignancies by abrogating tumor-induced angiogenesis . This may be accomplished by inhibiting the kinase activity of P15692 receptor-2 ( P35968 ) , which has a key role in mediating P15692 -induced responses . The novel indole-ether quinazoline DB04849 is a highly potent ( IC50 < 1 nmol/L ) DB00171 -competitive inhibitor of recombinant P35968 tyrosine kinase in vitro . Concordant with this activity , in human umbilical vein endothelial cells , DB04849 inhibited P15692 -stimulated proliferation and P35968 phosphorylation with IC50 values of 0.4 and 0.5 nmol/L , respectively . In a fibroblast/endothelial cell coculture model of vessel sprouting , DB04849 also reduced vessel area , length , and branching at subnanomolar concentrations . Once-daily oral administration of DB04849 ablated experimental ( P15692 -induced ) angiogenesis in vivo and inhibited endochondral ossification in bone or corpora luteal development in ovary ; physiologic processes that are highly dependent upon neovascularization . The growth of established human tumor xenografts ( colon , lung , prostate , breast , and ovary ) in athymic mice was inhibited dose-dependently by DB04849 , with chronic administration of 1.5 mg per kg per day producing statistically significant inhibition in all models . A histologic analysis of Calu-6 lung tumors treated with DB04849 revealed a reduction in microvessel density within 52 hours that became progressively greater with the duration of treatment . These changes are indicative of vascular regression within tumors . Collectively , the data obtained with DB04849 are consistent with potent inhibition of P15692 signaling , angiogenesis , neovascular survival , and tumor growth . DB04849 is being developed clinically as a once-daily oral therapy for the treatment of cancer . Interplay between O75030 , Q9Y6X2 , and P40763 in mast cells and melanocytes . Microphthalmia transcription factor ( O75030 ) and P40763 are two transcription factors that play a major role in the regulation of growth and function in mast cells and melanocytes . In the present study , we explored the O75030 - Q9Y6X2 - P40763 network of interactions , how these interactions regulate gene expression , and how cytokine-mediated phosphorylation of O75030 and P40763 is involved in the in vivo interplay between these three proteins . In NIH 3T3 cells stimulated via P40189 receptor , transfected O75030 was found to be phosphorylated at S409 . Such phosphorylation of O75030 leads to Q9Y6X2 dissociation from O75030 and its association with P40763 . Activation of mouse melanoma and mast cells through P40189 or c-Kit receptors induced the mobilization of Q9Y6X2 from O75030 to P40763 . In mast cells derived from O75030 (di/di) mice , whose O75030 lacks the Zip domain ( Q9Y6X2 -binding domain ) , we found downregulation in mRNA levels of genes regulated by either O75030 or P40763 . This regulatory mechanism is of considerable importance since it is likely to advance the deciphering of a role for O75030 and P40763 in mast cells and melanocytes . Thrombin receptors and their antagonists : an update on the patent literature . IMPORTANCE OF THE FIELD : Thrombin plays a central role in cardiovascular inflammation . Most of the cellular responses to thrombin are mediated by cell surface protease-activated receptors ( PARs ) . Several preclinical studies indicate that PARs are potential targets for treating cardiovascular diseases such as thrombosis , atherosclerosis and restenosis . Among PARs , P25116 has emerged as an important therapeutic target . AREAS COVERED IN THIS REVIEW : This review covers recent advances in the development of thrombin receptors antagonists . It is focused on the search for P25116 antagonists as this is at the moment the most promising and attractive target . However , some early promising studies on PAR-3 and -4 antagonists are also reported . WHAT THE READER WILL GAIN : The review has been written in order to give to the reader hints and references that cover , in our opinion , the most interesting and/or promising approaches in this research field . TAKE HOME MESSAGE : Research on P25116 antagonists has finally led to good clinical candidates such as DB05692 ( Schering-Plough ) and E-5555 ( Eisai Co. ) . Clinical trials clearly demonstrate that development of PAR1 antagonists is not only possible but most likely will lead to development of antiplatelet drugs as well as of drugs useful for the treatment of inflammatory , proliferative and neurodegenerative diseases . Migration potential and gene expression profile of human DB05914 induced by O15444 . Recruitment of DB05914 ( O60682 ) to tissue damages is a promising approach for in situ tissue regeneration . The physiological mechanisms and regulatory processes of O60682 trafficking to injured tissue remain poorly understood . However , the pivotal role of chemokines in O60682 recruitment has already been shown . The aim of this study was to determine the migratory potential and the gene expression profile of O60682 stimulated with the CC chemokine O15444 ( O15444 ) . Bone marrow derived human O60682 were exposed to different doses of O15444 in a standardized chemotaxis assay . Microarray gene expression profiling and pathway analysis were performed for O15444 stimulated O60682 . Maximum migration of O60682 towards O15444 was observed at 10(3) nM . Microarray analysis revealed an induction of molecules directly involved in chemotaxis and homing of bone marrow cells ( P09341 -3 , P10145 , Q07343 ) , cytoskeletal and membrane reorganisation ( P10145 , Q13393 , P08833 ) , cellular polarity ( Q13393 ) , and cell movement ( P09341 -3 , P80162 , P10145 , P35354 , Q07343 , P21980 ) . Respective chemokine secretion was confirmed by protein membrane-array analysis . The activation of P25025 ligands ( P09341 -3 , P42830 -6 , P10145 ) and a P15018 -receptor/ P40189 ligand ( P15018 ) indicated an involvement of the respective signaling pathways during initiation of chemotaxis and migration . These results suggest O15444 as a new potential candidate for further in situ regeneration approaches . Tetramethoxychalcone , a chalcone derivative , suppresses proliferation , blocks cell cycle progression , and induces apoptosis of human ovarian cancer cells . In the present study , we investigated the in vitro antitumor functions of a synthetic chalcone derivative 4,3',4',5'- tetramethoxychalcone ( TMOC ) in ovarian cancer cells . We found that TMOC inhibited the proliferation and colony formation of cisplatin sensitive cell line A2780 and resistant cell line A2780/ DB00515 , as well as ovarian cancer cell line SKOV3 in a time- and dose-dependent manner . Treatment of A2780 cells with TMOC resulted in G0/ P55008 cell cycle arrest through the down-regulation of cyclin D1 and P11802 , and the up-regulation of p16 , P38936 and p27 proteins . We demonstrated that TMOC might induce cell apoptosis through suppressing Bcl-2 and Bcl-xL , but enhancing the expression of Bax and the cleavage of P09874 . Treatment of TMOC also reduced the invasion and migration of A2780 cells . Finally , we found that TMOC inhibited the constitutive activation of P40763 signaling pathway and induced the expression of the tumor suppressor P60484 regardless of the p53 status in cell lines . These data suggest that TMOC may be developed as a potential chemotherapeutic agent to effectively treat certain cancers including ovarian cancer . Cyclic nucleotide phosphodiesterases ( PDEs ) in human osteoblastic cells ; the effect of PDE inhibition on DB02527 accumulation . The regulation of the secondary messengers , cyclic adenosine monophosphate ( DB02527 ) and cyclic guanosine monophosphate ( cGMP ) , is crucial in the hormonal regulation of bone metabolism . Both DB02527 and cGMP are inactivated by cyclic nucleotide phosphodiesterases ( PDEs ) , a superfamily of enzymes divided into 11 families ( PDE1-11 ) . We compared the PDEs of cultured human osteoblasts ( NHOst ) and SaOS-2 osteosarcoma cells . The PDE activity of NHOst cells consisted of PDE1 , PDE3 and PDE7 , whereas PDE1 , PDE7 and DB05876 , but no PDE3 activity was detected in SaOS-2 cells . In line with the difference in the PDE profiles , rolipram , a DB05876 inhibitor , increased the accumulation of DB02527 in SaOS-2 , but not in NHOst cells . Expression of PDE subtypes Q14123 , Q14432 , P27815 , Q07343 , Q13946 and Q9NP56 was detected in both cell types . NHOst cells additionally expressed P54750 . The endocannabinoid system -- back to the scene of cardiometabolic risk factors control ? This review examines the impact of the endocannabinoid signaling system on metabolic and cardiovascular health and the new therapeutic strategies that selectively target dysfunctional endocannabinoid action in peripheral tissues , without causing the undesirable central nervous system effects that occurred with the first-generation of P21554 receptor blockers . We first review the components of the endocannabinoid system and the enzymes that synthesize and degrade the endocannabinoids , the critical role of the system in the homeostasis of energy balance , and its hedonic aspects related to the incentive and motivational value of food . Second , we describe the central and peripheral actions of the endocannabinoid system and its interactions with other biological modulators , such as ghrelin and leptin . Third , we summarize data from human clinical trials with the P21554 inverse agonist rimonabant , showing that the drug , although effective in increasing weight loss with accompanying improvements in the metabolic profile of the participants in the DB03615 ( DB06155 In Obesity ) trials , was withdrawn from the market because of the risk of serious adverse events . Finally , we describe : 1 ) the development of new selective peripheral blockers that interrupt endocannabinoid action selectively in peripheral tissues and that have been suggested as an alternative approach to treat the metabolic consequences of obesity and related diseases , without undesirable central nervous system effects , and 2 ) the potential for inhibition of enzymes of synthesis , as well as the possible role of endocannabinoid congeners , with opposing effects as compared to P21554 receptor agonists , in the control of metabolic 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 . 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 . Guggulsterone inhibits angiogenesis by blocking P40763 and P15692 expression in colon cancer cells . The plant sterol guggulsterone has been shown to exert anti-tumor effects , making it a candidate chemotherapeutic agent . We investigated the anti-tumor effects of guggulsterone on colon cancer cells and elucidated the underlying molecular mechanisms related to angiogenesis . The apoptotic effects of guggulsterone were examined by cell survival assay . Western blot analysis was used to determine the levels of various down-stream intracellular proteins involved in angiogenesis , including signal transducer and activator of transcription 3 ( P40763 ) , vascular endothelial growth factor ( P15692 ) , hypoxia-inducible factor-1alpha ( HIF-1alpha ) and aryl hydrocarbon receptor nuclear translocator ( P27540 ) . Using chromatin immunoprecipitation assay , we tested whether guggulsterone affects the recruitment of P40763 , P27540 and HIF-1alpha to the human P15692 promoter . To investigate the effect of guggulsterone on vascular endothelial cell migration and invasion , tube formation and migration assays were conducted using human umbilical vein endothelial cells ( HUVECs ) . Matrix metalloproteinase ( MMP ) -2 and -9 activities were measured by gelatin zymography . Guggulsterone significantly reduced cell viability in colon cancer cells in a dose-dependent manner and blocked P15692 , P27540 and P40763 expression prominently in hypoxic conditions . The recruitment of P40763 and P27540 , but not HIF-1alpha , to the P15692 promoter was inhibited by guggulsterone treatment . HUVECs produced much foreshortened and severely broken tubes and showed decreased migration activity under guggulsterone effects . In addition , zymography revealed that P08253 and -9 enzyme activities were markedly lower in the presence of guggulsterone . The results of this study suggest that guggulsterone not only induces apoptosis , but also inhibits angiogenesis and metastasis in colon cancer cells by blocking P40763 and P15692 expression , suggesting its therapeutic potential in the treatment of colorectal cancer . New perspectives : role of sunitinib 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 gastrointestinal stromal tumour ( GIST ) . DB01268 is able to inhibit RTKs such as receptors for platelet-derived growth factor ( PDGF-R alpha and beta ) and for vascular endothelial growth factor ( VEGFRs ) . It is able to inhibit P10721 receptor , colony stimulating factor type 1 receptor ( P04141 - 1R ) , 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 sunitinib targets play important roles in growth and survival of human breast cancer ( BC ) . The " rationale " of sunitinib 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 sunitinib in metastatic BC . 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 . 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 . P40763 -mediated coincidence detection regulates noncanonical immediate early gene induction . Signaling pathways interact with one another to form dynamic networks in which the cellular response to one stimulus may depend on the presence , intensity , timing , or localization of other signals . In rare cases , two stimuli may be simultaneously required for cells to elicit a significant biological output . This phenomenon , generally termed " coincidence detection , " requires a downstream signaling node that functions as a Boolean AND gate to restrict biological output from a network unless multiple stimuli are received within a specific window of time . Simultaneous activation of the P01133 receptor ( P00533 ) and a thrombin receptor ( protease-activated receptor-1 , P25116 ) increases the expression of multiple immediate early genes ( IEGs ) associated with growth and angiogenesis . Using a bioinformatic comparison of IEG promoter regions , we identified P40763 as a critical transcription factor for the detection of coincident P00533 / P25116 activation . P00533 activation induces classical P40763 DB00135 (705) phosphorylation but also initiates an inhibitory signal through the PI3K-AKT signaling axis that prevents P40763 DB00133 (727) phosphorylation . Coincident P25116 signaling resolves these conflicting P01133 -activated pathways by blocking AKT activation and permitting GSK-3α/β-dependent P40763 DB00133 (727) phosphorylation and P40763 -dependent gene expression . Functionally , combinatorial P00533 / P25116 signaling suppresses P01133 -induced proliferation and thrombin-induced leukocyte adhesion and triggers a P40763 -dependent increase in endothelial cell migration . This study reveals a novel signaling role for P40763 in which the simultaneous presence of extracellular P01133 and thrombin is detected at the level of P40763 post-translational modifications . Collectively , our results describe a novel regulatory mechanism in which combinatorial P00533 / P25116 signaling regulates P40763 -dependent IEG induction and endothelial cell migration . 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 . 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 . A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors . P10721 or alpha-platelet-derived growth factor receptor ( alpha- P09619 ) activating mutations are the pathogenic mechanisms that characterize gastrointestinal stromal tumors ( GIST ) . Despite excellent responses to imatinib mesylate ( IM ) , patients are relapsing . We developed an IM-resistant GIST cell line ( GIST-R ) from the IM-sensitive GIST882 cell line ( GIST-S ) by growing these cells in IM . Gene expression profiling ( GEP ) of GIST-S , GIST-R cells and two IM resistant GIST patients demonstrated that P10721 is downregulated implying a major role in IM resistance . Instead , GIST-R cells have acquired IM resistance by overexpressing the oncogenic receptor tyrosine kinase - P30530 - in a ' kinase switch ' . Further , the two IM resistant GIST patients express P30530 and not c-Kit , seen by immunohistochemistry ( IHC ) . Real time reverse transcriptase-polymerase chain reaction and Western blotting of the GIST-S and GIST-R cells confirmed the switch from Kit to P30530 . In GIST-R , P30530 is tyrosine phosphorylated and its ligand growth-arrest-specific gene 6 is overexpressed implying autocrine activation . The kinase switch is associated with a morphological change from spindle to epithelioid . Molecular modeling of the kinase domain of mutant c-Kit ( V654A ) and P30530 showed no binding to IM but efficient binding to DB05216 , a novel c-Kit/ P30530 kinase inhibitor . DB05216 synergizes with docetaxel ( taxotere ) and is cytotoxic to GIST cells . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD .

MH_train_1160

MH_train_1160

MH_train_1160

interacts_with DB00622?

multiple_choice

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 . The anti-inflamm-aging and hepatoprotective effects of huperzine A in D-galactose-treated rats . Oxidative stress contributes to a chronic inflammatory process referred to as " inflamm-aging " . P22303 inhibitors ( AChEI ) can enhance cholinergic transmission and act as anti-inflammatory agents via immunocompetent cells expressing α-7 acetylcholine receptors ( AChR ) . The present study explores the possible role of huperzine A , a reversible and selective AChEI , against D-gal-induced oxidative damage , cell toxicity and inflamm-aging in rat livers . In two-month-old rats with normal liver function , an 8-week administration of D-gal ( 300 mg/kg subcutaneously ( s.c. ) injected ) , significantly increased hepatic impairment , ROS generation and oxidative damage , hepatic senescence , nuclear factor-kappa B ( NF-κB ) activation and inflammatory responses . An 8-week co-administration of both D-gal ( 300 mg/kg s.c. ) and huperzine A ( 0.1 mg/kg s.c. ) not only significantly decreased hepatic function impairment , ROS generation , oxidative damage , but also suppressed inflamm-aging by inhibiting hepatic replicative senescence , P22303 activity , IκBα degradation , NF-κB p65 nuclear translocation and inflammatory responses . The expression levels of pro-inflammatory cytokine mRNA and proteins , such as TNFα , IL-1β and P05231 decrease significantly , and the protein levels of the anti-inflammatory cytokine P22301 display an obvious increase . These findings indicated that D-gal-induced hepatic injury and inflamm-aging in the rat liver was associated with the development of a pro-inflammatory phenotype in this organ . D-gal induced damage-associated molecular patterns ( DAMPs ) because oxidative damages might play an important role in D-gal-induced hepatic sterile inflammation . DB04864 exhibited protective effects against D-gal-induced hepatotoxicity and inflamm-aging by inhibiting P22303 activity and via the activation of the cholinergic anti-inflammatory pathway . The huperzine A mechanism might be involved in the inhibition of DAMPs-mediated NF-κB nuclear localization and activation . Molecular and biochemical analysis of calmodulin interactions with the calmodulin-binding domain of plant glutamate decarboxylase . We previously provided what to our knowledge is the first evidence that plant glutamate decarboxylase ( Q99259 ) is a calmodulin ( P62158 ) -binding protein . Here , we studied the Q99259 P62158 -binding domain in detail . A synthetic peptide of 26 amino acids corresponding to this domain forms a stable complex with Ca2+/ P62158 with a 1:1 stoichiometry , and amino acid substitutions suggest that tryptophan-485 has an indispensable role in P62158 binding . Chemical cross-linking revealed specific P62158 / Q99259 interactions even in the absence of Ca2+ . However , increasing KCI concentrations or deletion of two carboxy-terminal lysines abolished these interactions but had a mild effect on P62158 / Q99259 interactions in the presence of Ca2+ . We conclude that in the presence of Ca(2+)-hydrophobic interactions involving tryptophan-485 and electrostatic interactions involving the carboxy-terminal lysines mediate P62158 / Q99259 complex formation . By contrast , in the absence of Ca2+ , P62158 / Q99259 interactions are essentially electrostatic and involve the carboxy-terminal lysines . In addition , a tryptophan residue and carboxy-terminal lysines are present in the P62158 -binding domain of an Arabidopsis Q99259 . Finally , we demonstrate that petunia Q99259 activity is stimulated in vitro by Ca2+/ P62158 . Our study provides a molecular basis for Ca(2+)-dependent P62158 / Q99259 interactions and suggests the possible occurrence of Ca(2+)-independent P62158 / Q99259 interactions . Cortical parvalbumin GABAergic deficits with α7 nicotinic acetylcholine receptor deletion : implications for schizophrenia . Dysfunction of cortical parvalbumin ( PV ) -containing GABAergic interneurons has been implicated in cognitive deficits of schizophrenia . In humans microdeletion of the P36544 ( α7 nicotinic acetylcholine receptor , nAChR ) gene is associated with cortical dysfunction in a broad spectrum of neurodevelopmental and neuropsychiatric disorders including schizophrenia while in mice similar deletion causes analogous abnormalities including impaired attention , working-memory and learning . However , the pathophysiological roles of α7 nAChRs in cortical PV GABAergic development remain largely uncharacterized . In both in vivo and in vitro models , we identify here that deletion of the α7 nAChR gene in mice impairs cortical PV GABAergic development and recapitulates many of the characteristic neurochemical deficits in PV-positive GABAergic interneurons found in schizophrenia . α7 nAChR null mice had decreased cortical levels of GABAergic markers including PV , glutamic acid decarboxylase 65/67 ( Q05329 /67 ) and the α1 subunit of GABAA receptors , particularly reductions of PV and Q99259 levels in cortical PV-positive interneurons during late postnatal life and adulthood . Cortical GABAergic synaptic deficits were identified in the prefrontal cortex of α7 nAChR null mice and α7 nAChR null cortical cultures . Similar disruptions in development of PV-positive GABAergic interneurons and perisomatic synapses were found in cortical cultures lacking α7 nAChRs . Moreover , DB01221 receptor expression was reduced in GABAergic interneurons , implicating DB01221 receptor hypofunction in GABAergic deficits in α7 nAChR null mice . Our findings thus demonstrate impaired cortical PV GABAergic development and multiple characteristic neurochemical deficits reminiscent of schizophrenia in cortical PV-positive interneurons in α7 nAChR gene deletion models . This implicates crucial roles of α7 nAChRs in cortical PV GABAergic development and dysfunction in schizophrenia and other neuropsychiatric disorders . Absence of clinically important Q12809 channel blockade by three compounds that inhibit phosphodiesterase 5 -- sildenafil , tadalafil , and vardenafil . Compounds that inhibit phosphodiesterase 5 ( O76074 ) have been developed for the treatment of erectile dysfunction . Because men with erectile dysfunction frequently have comorbid cardiovascular disease , they may have limited cardiac repolarization reserve and be at risk of arrhythmia if treated with medications that prolong ventricular repolarization . The human ether-a-go-go related gene ( Q12809 ) channel is important for repolarization in human myocardium and is a common target for drugs that prolong the QT interval . We studied the ability of three compounds that inhibit O76074 -- sildenafil , tadalafil , and vardenafil -- to block the Q12809 channel . Using a whole cell variant of the patch-clamp method , the Q12809 current was measured in a stably transfected human embryonic kidney cell line expressing the Q12809 channel . The compounds produced dose-dependent reductions in Q12809 current amplitude over a concentration range of 0.1 to 100 microM . The IC50 values were 12.8 microM for vardenafil and 33.3 microM for sildenafil . Because the maximum soluble concentration of tadalafil ( 100 microM ) produced only a 50.9 % inhibition of the Q12809 current amplitude , the IC50 value for tadalafil could not be determined with the Hill equation . DB00820 had the weakest capacity to block the Q12809 channel , producing a 50.9 % blockade at the maximum soluble concentration ( 100 microM ) , compared with 86.2 % for vardenafil ( 100 microM ) and 75.2 % for sildenafil ( 100 microM ) . In conclusion , the concentrations of the O76074 inhibitors required to evoke a 50 % inhibition of the Q12809 current were well above reported therapeutic plasma concentrations of free and total compound . None of the three compounds was a potent blocker of the Q12809 channel . Synergistic antileukemic effects between DB06080 and chemotherapy involve downregulation of cell cycle-regulated genes and c-Mos-mediated MAPK pathway . Internal tandem duplications ( ITDs ) of fms-like tyrosine kinase 3 ( P36888 ) receptor play an important role in the pathogenesis of acute myeloid leukemia ( AML ) and represent an attractive therapeutic target . DB06080 has demonstrated potent effects in AML cells with P36888 -ITDs . Here , we provide further evidence that DB06080 treatment significantly downregulates cyclins D and E but increases the expression of P38936 and p27 . DB06080 induces apoptosis through downregulation of Bcl-xL and upregulation of Q16611 , P55957 and Q92934 . We also evaluate the combinations of DB06080 and chemotherapy . DB06080 demonstrates significant sequence-dependent synergism with cytarabine and doxorubicin in cell lines and primary leukemia samples . The optimal combination was validated in MV4-11 xenografts . Low-density array analysis revealed the synergistic interaction involved in downregulation of cell cycle and mitogen-activated protein kinase pathway genes . P24385 and c-Mos were the most significantly inhibited targets on both transcriptional and translational levels . Treatment with short hairpin RNAs targeting either P24385 or c-Mos further sensitized MV4-11 cells to DB06080 . These findings suggest that specific pathway genes were further targeted by adding chemotherapy and support the rationale of combination therapy . Thus , a clinical trial using sequence-dependent combination therapy with DB06080 in AML is warranted . Computational modeling of the direct hydride transfer mechanism for the MAO catalyzed oxidation of phenethylamine and benzylamine : ONIOM ( QM/QM ) calculations . Monoamine oxidases are two isozymic flavoenzymes which are the important targets for drugs used in the treatment of depression , Parkinson and Alzheimer 's diseases . The catalytic reaction taking place between the cofactor DB03147 and amine substrate is still not completely understood . Herein we employed quantum chemical methods on the recently proposed direct hydride transfer mechanism including full active site residues of MAO isoforms in the calculations . Activation free energy barriers of direct hydride transfer mechanism for P21397 and P27338 were calculated by ONIOM ( our own n-layered integrated molecular orbital + molecular mechanics ) method with QM/QM ( quantum mechanics:quantum mechanics ) approach employing several density functional theory functionals , B3LYP , WB97XD , P62158 -B3LYP and M06-2X , for the high layer . The formation of very recently proposed αC-flavin N5 adduct inside the enzyme has been investigated . ONIOM ( M06-2X/6-31+G(d,p):PM6 ) results revealed that such an adduct may form only in P27338 suggesting slightly different hydride transfer mechanisms for P21397 and P27338 . 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 . Effects of the dual Q07869 -α/γ agonist aleglitazar on glycaemic control and organ protection in the Zucker diabetic fatty rat . AIMS : To evaluate the effects of aleglitazar , a dual peroxisome proliferator-activated receptor-α/γ agonist , on the development of diabetes-related organ dysfunction , in relation to glycaemic and lipid changes , in Zucker diabetic fatty ( ZDF ) rats . METHODS : Six-week-old , male ZDF rats received aleglitazar 0.3 mg/kg/day or vehicle as food admix for 13 weeks ( n = 10 per group ) . Age-matched male Zucker lean rats served as non-diabetic controls . Plasma and renal markers were measured at several time points . Histopathology and quantitative immunohistochemistry were performed at 13 weeks . RESULTS : Glycated haemoglobin ( 5.4 vs. 9.2 % ) and blood glucose ( 8.3 ± 0.3 vs. 26.1 ± 1.0 mmol/l ) were significantly reduced at 12 weeks with aleglitazar versus vehicle-treated ZDF rats ( both p < 0.01 ) , while aleglitazar preserved near-normal plasma insulin levels . DB08915 prevented the development of hypertriglyceridaemia ( 1.4 ± 0.1 vs. 8.5 ± 0.9 mmol/l ) and reduced plasma non-esterified fatty acids ( 0.09 ± 0.02 vs. 0.26 ± 0.04 mmol/l ) relative to vehicle-treated animals ( both p < 0.01 ) . Urinary glucose and protein concentrations were significantly reduced at 13 weeks with aleglitazar versus vehicle-treated rats ( both p < 0.01 ) . Consistent with its effect on glycaemic control , aleglitazar protected β-cell morphology , as evidenced by preservation of islet integrity , and reduction of β-cell apoptosis and islet fibrosis . DB08915 prevented renal glomerular hypertrophy , podocyte degeneration , glomerulosclerosis , tubulo-interstitial lesions and development of cataracts . CONCLUSIONS : DB08915 strongly improved glycaemic and lipid parameters while protecting key tissues , including the pancreas , kidneys and eyes , against diabetes-associated structural and functional changes in the ZDF rat . Role of heme oxygenase-1 in protection of the kidney after hemorrhagic shock . Hemorrhagic shock followed by resuscitation ( HSR ) causes oxidative stress , which results in multiple organ damage . The kidney is one of the target organs of HSR-mediated oxidative tissue injury . Heme oxygenase ( HO ) -1 , the rate-limiting enzyme in heme catabolism , is induced by oxidative stress ; it protects against oxidative tissue injuries . The aim of the present study was to examine the role of renal P09601 induction after HSR . Rats were subjected to hemorrhagic shock to achieve a mean arterial pressure of 30 mmHg for 60 min , followed by resuscitation with the shed blood . HSR resulted in a significant increase in functional P09601 protein in the tubular epithelial cells of the kidney , whereas HSR resulted in only a slight increase in gene expression of tumor necrosis factor ( P01375 ) -alpha and inducible nitric oxide synthase ( P35228 ) , and in protein expression of activated caspase-3 solely in renal cells where P09601 expression was absent . HSR also resulted in a significant increase in Bcl-2 gene expression . Pretreatment of HSR animals with DB04912 ( 0.5 micromol/kg ) , a specific competitive inhibitor of HO activity , resulted in a significant decrease in HO activity and exacerbated tissue inflammation and apoptotic cell death as judged by the marked increase in expression of P01375 and P35228 , and in activated caspase-3-positive cells , and the significant reduction in Bcl-2 expression , respectively . These findings indicate that P09601 induction is an adaptive response to HSR-induced oxidative stress and is essential for protecting tubular epithelial cells from oxidative damage through its anti-inflammatory and anti-apoptotic properties . Role of chronic inhibition of dopamine-metabolizing enzymes in the regulation of renal sodium and phosphate excretion in the rat remnant kidney . BACKGROUND/AIMS : The present study examined the effects of chronic selective or combined inhibition of type A monoamine oxidase ( MAO ) and catechol-O-methyltransferase ( P21964 ) on daily urinary excretion of dopamine and metabolites and on natriuresis and phosphaturia in 3/4 nephrectomized ( 3/4nx ) and Sham rats . METHODS : The 3/4nx and Sham rats were placed in metabolic cages and received the P21397 -selective inhibitor Ro-411049 ( 7.5 mg x kg(-1) bid ) and/or the P21964 -selective inhibitor DB03336 3-202 ( 30 mg x kg(-1) bid ) orally for 3 days during high sodium diet . RESULTS : Selective P21964 inhibition increased the urinary excretion of the deaminated metabolite ( 3,4-dihydroxyphenylacetic acid , DOPAC ) and decreased the urinary excretion of the methylated ( 3-methoxytyramine , 3-MT ) and deaminated plus methylated metabolite ( homovanillic acid , HVA ) in both groups . Selective P21397 inhibition increased the urinary excretion of 3-MT and reduced the urinary excretion of both DOPAC and HVA in either 3/4nx or Sham rats . Combined inhibition of P21397 and P21964 did not significantly change the urinary excretion of DOPAC and markedly decreased the urinary excretion of 3-MT and HVA in both groups . Selective or combined inhibition of P21397 and P21964 did not alter the daily urinary excretion of dopamine , sodium or phosphate in either 3/4nx or Sham rats . CONCLUSIONS : Chronic selective or combined inhibition of P21397 and P21964 is not of major importance in regulating the dopamine-dependent natriuresis and phosphaturia in either 3/4nx or Sham rats . 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 . Expression of markers for transitional cell carcinoma in normal bladder mucosa of patients with bladder cancer . PURPOSE : Because we have found that random mucosal biopsies have no additional prognostic value to conventional histopathology , we studied biopsies of histologically normal bladder mucosa with several molecular markers believed to be associated with the development of transitional cell carcinoma . MATERIALS AND METHODS : Six groups of patients with an increasing stage of bladder tumor were selected : ( 1 ) benign disease ( for example , benign prostatic hyperplasia , n = 8 ) ; ( 2-4 ) low ( n = 10 ) , intermediate ( n = 9 ) and high risk ( n = 7 ) superficial tumors ; ( 5 ) progressive superficial tumors resistant to optimal conservative therapy ( n = 6 ) ; ( 6 ) invasive or disseminated tumors at presentation ( n = 5 ) . We studied the expression of cytokeratin ( used as an epithelial marker ) , fibronectin , P12830 ( HECD-1 ) , I- P62158 , human leukocyte antigen ( HLA ) -I , HLA-II and epidermal growth factor receptor ( P01133 -R ) in cold-cup biopsies of normal mucosa . RESULTS : P02751 , HECD-1 , I- P62158 and HLA-II expression showed no significant changes in the different groups . There was a significant increase in the expression of HLA-I ( p = .003 ) and P01133 -R ( p = .0001 ) with a higher stage of the tumor . CONCLUSION : An increasing P01133 -R expression in normal looking mucosa of patients with increasing stages of bladder tumors could be a prognostic factor or might indicate that this increase in expression is not tumor specific but is seen in the whole bladder . 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 . Effects of an alpha 7-nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3-(2,4-dimethoxybenzylidene)-anabaseine ( DB05708 ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 , the α7-nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 genotype . CONCLUSIONS : The observed DB05708 -related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7-nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents . Design and synthesis of 3,5-disubstituted-1,2,4-oxadiazoles as potent inhibitors of phosphodiesterase4b2 . A series of 3,5-disubstituted-1,2,4-oxadiazoles has been prepared and evaluated for phosphodiesterase inhibition ( PDE4B2 ) . Among the prepared 3,5-disubstituted-1,2,4-oxadiazoles , compound 9a is the most potent inhibitor ( PDE4B2 IC(50) = 5.28 μm ) . Structure-activity relationship studies of 3,5-disubstituted-1,2,4-oxadiazoles revealed that substituents 3-cyclopentyloxy-4-methoxyphenyl group at 3-position and cyclic ring bearing heteroatoms at 5-position are important for activity . Molecular modeling study of the 3,5-disubstituted-1,2,4-oxadiazoles with Q07343 has shown similar interactions of 3-cyclopentyloxy-4-methoxyphenyl group ; however , heteroatom ring is slightly deviating when compared to DB01791 . 3-(3-Cyclopentyloxy-4-methoxyphenyl)-5-(piperidin-4-yl)-1,2,4-oxadiazole ( 9a ) exhibited good analgesic and antiinflammatory activities in formalin-induced pain in mice and carrageenan-induced paw edema model in rat . 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 . P13688 impedes thyroid cancer growth but promotes invasiveness : a putative mechanism for early metastases . P13688 , also known as biliary glycoprotein ( BGP ) , CD66a , Q00839 and C- P62158 , is a member of the P06731 immunoglobulin superfamily . P13688 is a putative tumor suppressor based on diminished expression in some solid neoplasms such as colorectal carcinoma . However , P13688 is overexpressed in some tumors such as non-small cell lung cancer . To clarify the mechanism of action of this cell adhesion molecule , we studied thyroid carcinoma that has a spectrum of morphologies and variable behavior allowing separation of proliferation from invasion and metastasis . P13688 is expressed in thyroid carcinoma cell lines derived from tumors that exhibit aggressive behavior . Introduction of P13688 into endogenously deficient WRO cells resulted in reduced cell cycle progression associated with P38936 upregulation and diminished Rb phosphorylation . Forced P13688 expression enhanced cell-matrix adhesion and migration and promoted tumor invasiveness . Conversely , small interfering RNA ( siRNA ) -mediated downregulation of P13688 expression in Q9BYG7 cells accelerated cell cycle progression and significantly enhanced tumor size in xenografted mice . P13688 is not appreciably expressed in normal thyroid tissue or benign thyroid tumors . In a human thyroid tissue array , P13688 reactivity was associated with metastatic spread but not with increased tumor size . These findings identify P13688 as a unique mediator that restricts tumor growth whereas increasing metastatic potential . Our data highlight a complex repertoire of actions providing a putative mechanism underlying the spectrum of biologic behaviors associated with thyroid cancer . Genomic structure and chromosome location of the murine Q01064 phosphodiesterase gene . Cyclic nucleotide phosphodiesterases ( PDEs ) catalyze the hydrolysis of DB02527 and cGMP , thereby participating in regulation of the intracellular concentrations of these second messengers . The PDE1 family is defined by regulation of activity by calcium and calmodulin . We have cloned and characterized the mouse Q01064 gene , which encodes the 63-kDa calcium/calmodulin-dependent PDE ( P62158 -PDE ) , an isozyme that is expressed in the CNS in the olfactory tract , dentate gyrus , and striatum and may participate in learning , memory , and regulation of phosphorylation of Q9UD71 in dopaminergic neurons . We screened an I-129/SvJ mouse genomic library and identified exons 2-13 of the Q01064 gene that span 8.4 kb of genomic DNA . Exons range from 67 to 205 nucleotides and introns from 91 to 2250 nucleotides in length . Exon 1 was not present in the 3 kb of genomic DNA 5' to exon 2 in our clones . The mouse Q01064 gene shares many similar or identical exon boundaries as well as considerable sequence identity with the rat Q07343 and Q08499 genes and the Drosophila dunce DB02527 -specific PDE gene dnc , suggesting that these genes all arose from a common ancestor . Using fluorescence in situ hybridization , we localized the Q01064 gene to the distal tip of mouse Chromosome ( Chr ) 15 . DB08888 for vitreoretinal diseases . P02751 and laminin are clinically relevant plasmin receptors in the eye . Located at the vitreoretinal interface , they are cleaved by ocriplasmin ( DB05028 , ThromboGenics , Iselin , NJ ) , a novel ophthalmic medication . A series of clinical trials to study ocriplasmin for the treatment of vitreoretinal diseases such as vitreomacular traction , macular hole , and exudative age-related macular degeneration are underway . The results are promising and may impact patient care .

MH_train_1161

MH_train_1161

MH_train_1161

interacts_with DB04946?

multiple_choice

Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . Effect of type-selective inhibitors on cyclic nucleotide phosphodiesterase activity and insulin secretion in the clonal insulin secreting cell line BRIN-BD11 . 1. The cyclic nucleotide phosphodiesterases ( PDEs ) present in an insulin secreting cell line , BRIN - BD11 , were characterized using calcium/calmodulin , DB01277 , isoenzyme-selective PDE inhibitors and RT - PCR . 2 . P62158 activated cyclic AMP or cyclic GMP PDE activity in pellet and was 3 fold ( P=0.002 ) more potent in activating cyclic nucleotide hydrolysis in pellet compared with supernatant fractions . 3 . The PDE1/ O76074 inhibitor zaprinast inhibited both cyclic AMP and cyclic GMP PDE activity in both pellet and supernatant fractions of cell homogenates by a maximum of around 25 % ( IC(50) 1 - 5 microM ) , while rolipram ( DB05876 selective ) inhibited only cyclic AMP hydrolysis . 4 . The PDE3-selective inhibitors Org 9935 ( 0.02 - 10 microM ) and siguazodan ( 0.1 - 10 microM ) inhibited cyclic AMP PDE activity in the pellet but not the supernatant fractions of cell homogenates , with a maximum inhibition of about 30 % . DB01277 ( 2 - 7.5 ng ml(-1) ) potently augmented this PDE activity . 5 . RT - PCR using specific primers for Q13370 , but not for Q14432 , amplified , from BRIN - BD11 cell total RNA , a 351 base pair product that was > 97 % homologous with rat adipose tissue Q13370 . 6 . DB07954 , Org 9935 , siguazodan and rolipram ( 1 - 50 microM ) , but not zaprinast , each augmented glucose-induced insulin secretion in the presence of 16.7 mM but not 1 mM glucose . 7 . These findings , in a clonal insulin secreting cell line , are consistent with an important role for Q13370 in regulating the pool of cyclic AMP relevant to the modulation of glucose-induced insulin secretion . 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 . Both overlapping and distinct signaling pathways for somatostatin receptor subtypes P30872 and P30874 in pituitary cells . To elucidate the signaling events mediated by specific somatostatin receptor ( SSTR ) subtypes , we expressed P30872 and P30874 individually in rat pituitary GH12C1 and F4C1 cells , which lack endogenous somatostatin receptors . In transfected GH12C1 cells , both P30872 and P30874 coupled to inhibition of Ca2+ influx and hyperpolarization of membrane potential via a pertussis toxin ( PTx ) -sensitive mechanism . These effects reflected modulation of ion channel activities which are important for regulation of hormone secretion . Somatostatin analogs MK678 and CH275 acted as subtype selective agonists as expected . In transfected F4C1 cells , both P30872 and P30874 mediated somatostatin-induced inhibition of adenylyl cyclase via a PTx-sensitive pathway . In addition , activation of P30874 in F4C1 cells , but not P30872 , stimulated phospholipase C ( P98160 ) activity and an increase in [Ca2+]i due to release of Ca2+ from intracellular stores . Unlike adenylyl cyclase inhibition , the P98160 -mediated response was only partially sensitive to PTx . To determine the structural determinants in P30874 necessary for activation of P98160 , we constructed chimeric receptors in which domains of P30874 were introduced into P30872 . Chimeric receptors containing only the third intracellular loop , or all three intracellular loops from P30874 , mediated inhibition of adenylyl cyclase , but failed to stimulate P98160 activity as did wild-type P30874 . Furthermore , the C-terminal tail of P30874 was not required for coupling to P98160 . Thus , by expressing individual somatostatin receptor subtypes in pituitary cells , we have identified both overlapping and distinct signaling pathways for P30872 and P30874 , and have shown that sequences other than simply the intracellular domains are required for P30874 to couple to the P98160 signaling pathway . Protein kinases as drug targets in cancer . Identification of the key roles of protein kinases in signaling pathways leading to development of cancer has caused pharmacological interest to concentrate extensively on targeted therapies as a more specific and effective way for blockade of cancer progression . This review will mainly focus on inhibitors targeting these key components of cellular signaling by employing a technology-based point of view with respect to DB00171 - and non- DB00171 -competitive small molecule inhibitors and monoclonal antibodies of selected protein kinases , particularly , mammalian target of rapamycin ( P42345 ) , P11274 - P00519 , MEK , p38 MAPK , P00533 P09619 , VEGFR , P04626 and Raf . Inhibitors of the heat shock protein Hsp90 are also included in a separate section , as this protein plays an essential role for the maturation/proper activation of cancer-related protein kinases . In the following review , the molecular details of the mode of action of these inhibitors as well as the emergence of drug resistance encountered in several cases are discussed in light of the structural , molecular and clinical studies conducted so far . 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 . Stimulation of type 1 and type 8 Ca2+/calmodulin-sensitive adenylyl cyclases by the Gs-coupled 5-hydroxytryptamine subtype 5-HT7A receptor . The neurotransmitter serotonin ( 5-hydroxytryptamine , 5-HT ) plays an important regulatory role in developing and adult nervous systems . With the exception of the 5- Q9H205 receptor , all of the cloned serotonin receptors belong to the G protein-coupled receptor superfamily . Subtypes P50406 and P34969 couple to stimulation of adenylyl cyclases through Gs and display high affinities for antipsychotic and antidepressant drugs . In the brain , mRNA for P50406 is found at high levels in the hippocampus , striatum , and nucleus accumbens . P34969 mRNA is most abundant in the hippocampus , neocortex , and hypothalamus . To better understand how serotonin might control DB02527 levels in the brain , we coexpressed P50406 or 5-HT7A receptors with specific isoforms of adenylyl cyclase in P29320 293 cells . The P50406 receptor functioned as a typical Gs-coupled receptor in that it stimulated O95622 , a Gs-sensitive adenylyl cyclase , but not Q99440 or P40145 , calmodulin ( P62158 ) -stimulated adenylyl cyclases that are not activated by Gs-coupled receptors in vivo . Surprisingly , serotonin activation of 5-HT7A stimulated Q99440 and P40145 by increasing intracellular Ca2+ . 5-HT also increased intracellular Ca2+ in primary neuron cultures . These data define a novel mechanism for the regulation of intracellular DB02527 by serotonin . 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 . Isoenzyme-specific cyclooxygenase inhibitors : a whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6 . NSAIDs inhibit the conversion of arachidonic acid into DB03866 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase ( P36551 ) . Two genetically distinct isoforms have been discovered , P23219 and P35354 . While P23219 is thought to account for homeostatic amounts of eicosanoids , P35354 is induced during inflammation leading to pathologic amounts of eicosanoids . Since NSAIDs inhibit both P36551 isoforms , antiinflammatory drug research has refocused to discovering P35354 inhibitors that do not inhibit P23219 . For this purpose , we have developed a whole cell assay system using the human erythroleukemic cell line HEL as a source for P23219 and the human monocytic cell line Mono Mac 6 as a source for P35354 . Mono Mac 6 cells express high amounts of P35354 upon stimulation with lipopolysaccharide ( LPS ) in the absence of any detectable P23219 protein . On the other hand , we find HEL cells to naturally express P23219 protein , but not P35354 . Testing of a panel of NSAIDs as well as some P35354 specific inhibitors showed that this assay system is suitable for identifying compounds that selectively inhibit either P23219 or P35354 . This test system offers the advantage of assessing P23219 and P35354 inhibitors within the human species , within a similar test set-up , and circumvents the need for tedious purification of either platelets or peripheral blood monocytes . EPO receptor circuits for primary erythroblast survival . EPO functions primarily as an erythroblast survival factor , and its antiapoptotic actions have been proposed to involve predominantly P19957 -kinase and BCL-X pathways . Presently , the nature of EPO-regulated survival genes has been investigated through transcriptome analyses of highly responsive , primary bone marrow erythroblasts . Two proapoptotic factors , Bim and FoxO3a , were rapidly repressed not only via the wild-type P19235 , but also by PY-deficient knocked-in P19235 alleles . In parallel , Pim1 and Pim3 kinases and Irs2 were induced . For this survival gene set , induction failed via a PY-null P19235 -HM allele , but was restored upon reconstitution of a PY343 P42229 -binding site within a related P19235 -H allele . Notably , P19235 -HM supports erythropoiesis at steady state but not during anemia , while P19235 -H exhibits near wild-type P19235 activities . P19235 -H and the wild-type P19235 ( but not P19235 -HM ) also markedly stimulated the expression of Trb3 pseudokinase , and intracellular serpin , Serpina-3G . For SERPINA-3G and TRB3 , ectopic expression in EPO-dependent progenitors furthermore significantly inhibited apoptosis due to cytokine withdrawal . BCL-XL and P10415 also were studied , but in highly responsive Kit(pos)CD71(high)Ter119(neg) erythroblasts , neither was EPO modulated . P19235 survival circuits therefore include the repression of Bim plus FoxO3a , and P19235 /PY343/ P42229 -dependent stimulation of Pim1 , Pim3 , Irs2 plus Serpina-3G , and Trb3 as new antiapoptotic effectors . DB04894 labeled with Tc-99m for imaging tumors . DB04894 ( RC-160 ) , an octapeptide analog of somatostatin , has a high affinity for somatostatin receptor subtypes P30874 and P35346 . DB04894 binds differently to the tumors of the breast , ovary , exocrine pancreas , prostate and colon , than octreotide another octapeptide analog of somatostatin . DB04894 was labeled with Tc-99m , a radionuclide highly suitable for scintigraphic imaging . The labeling procedure was simple , produced > 70 % yields and could be applicable to label other peptides containing a cystine bridge . HPLC analysis showed that the tracer was stable when Tc-99m-RC-160 was challenged with 100 fold molar excess DTPA ( diethylenetriaminepentaacetic acid ) , HSA ( human serum albumin ) or cysteine and incubated at 37 degrees C for 4 h . HPLC analysis of urine samples obtained from mice that received Tc-99m-RC-160 showed that the preparation was stable in vivo . Rat brain cortex membrane receptor displacement assays showed that the Kd values for Tc-99m-RC-160 ( 71x10(-9) M ) and Tc-99m-octreotide ( 86x10(-9) M ) ( Sandostatin(R) ) were in nM range , and were similar to that for I-125-RC-160 ( 46x10(-9) M ) . High binding affinity of Tc-99m-RC-160 for human breast tumor cells SKBR-3 was also observed . These results suggest that Tc-99m-RC-160 is worthy of evaluation as an agent for scintigraphic imaging of tumors rich in somatostatin receptor subtypes P30874 and P35346 . 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 . Biological aspects of macrophage-stimulating protein ( MSP ) and its receptor . P26927 ( MSP ; also known as P14210 -like protein [ HGFl ] ) is a 78 kDa plasma protein that is secreted by the liver into the circulation as single-chain , biologically inactive pro-MSP . The presence of conserved triple disulfide loops ( kringles ) places pro-MSP in a family of coagulation system serine protease zymogens that are activated by proteolytic cleavage . Although pro-MSP has lost enzymic activity , it has retained the activation mechanism , in that proteolytic cleavage at a single site yields biologically active disulfide-linked alpha beta-chain heterodimeric MSP . The Q04912 is a transmembrane protein tyrosine kinase . MSP causes phosphorylation of the receptor cytoplasmic domain , association of phosphatidylinositol ( PI ) -3 kinase with the receptor , and phosphorylation of receptor-bound P19957 kinase . Inhibition of P19957 kinase by wortmannin prevents MSP action on cells . MSP stimulates motility of murine resident peritoneal macrophages . However , it does not act on exudate macrophages or blood monocytes , since these earlier maturational stages of the lineage do not express the receptor . MSP also stimulates keratinocyte cell lines , causing either chemotactic responses or increased cell numbers in culture . We suggest that pro-MSP diffuses into local tissue sites , where proteolytic cleavage to MSP results in stimulation of keratinocytes and macrophages . It possibly plays a role in tissue injury or wound healing . P04035 inhibition reduces the proinflammatory activation of human vascular smooth muscle cells by the terminal complement factor C5b-9 . The terminal complement complex C5b-9 is known to participate in inflammatory processes including atherosclerosis . Inflammation appears to be a direct consequence of C5b-9-mediated cell stimulation . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors may exert anti-inflammatory effects on vascular cells independent of lowering plasma cholesterol . Thus , we studied activation of vascular smooth muscle cells ( VSMCs ) by C5b-9 focusing on whether inhibition of the P04035 can reduce the proinflammatory effects of C5b-9.C5b-9 in sublytic concentrations increased the proliferation of human VSMCs and induced a time-dependent activation of the mitogen-activated protein ( Q96HU1 ) kinase extracellular signal-regulated kinase ( P29323 ) . Proliferation and P27361 /2 activation could be inhibited by the specific P29323 inhibitor PD98059 . HMG- DB01992 inhibition with cerivastatin-reduced VSMC proliferation and C5b-9-induced P27361 /2 activation . DB00439 also reduced the C5b-9-induced synthesis of the proinflammatory interleukin-6 ( P05231 ) . Furthermore , C5b-9 induced activation of the transcription factors activator protein- 1 ( AP-1 ) and nuclear factor-kappaB ( NF-kappaB ) , which could be inhibited by pretreatment of VSMCs with cerivastatin . L-mevalonate and geranylgeranylpyrophosphate reversed the inhibitory effects of cerivastatin . The present study in VSMCs shows that cerivastatin inhibits P05231 synthesis and cell proliferation induced by the terminal complement complex C5b-9 . This may be an important mechanism contributing to the beneficial effects of P04035 inhibitors beyond lowering of plasma cholesterol . [ 5-hydroxytryptamine ( serotonin ) receptors -- nomenclature and classification of types and subtypes ] . 5-HT receptors represent a superfamily of receptors with the largest known number of receptor subtypes . At present 15 receptor subtypes of three groups has been recognized . The 5-HT1 subfamily of receptors contains subtypes P08908 , P28222 , P28221 , P28566 , and P30939 ; activation of all of them results in the inhibition of adenylylcyclase . The subfamily of 5-HT2 contains subtypes 5- Q13049 , P41595 , and P28335 ; their activation leads to the stimulation of P98160 . Finally , subfamily of miscellaneous 5-HT receptors contains subtypes 5- Q9H205 , Q13639 , 5-HT5 , P50406 , and P34969 ; some of them has been cloned , however , our knowledge on their function is still minimal . 5-HT receptors participate in many physiological functions and a disturbance in serotonergic neurotransmission might cause several types of disease . 5-HT plays an important role in depression ; to cure this disease , drugs which increase levels of this neurotransmitter are used . A new drug group called Selective Serotonin Reuptake Inhibitors ( SSRI ) has been recently discovered . These drugs block the reuptake of 5-HT into nerve endings . There is an intensive search for new selective agonists as well as antagonists which could be use not only in the classification of receptor subtypes but which also possess certain therapeutic potential . Vinblastine rapidly induces Q13794 and acutely sensitizes primary chronic lymphocytic leukemia cells to ABT-737 . Proteins of the P10415 family provide a survival mechanism in many human malignancies , including chronic lymphocytic leukemia ( CLL ) . The P10415 inhibitor DB05764 ( navitoclax ) is active in clinical trials for lymphoid malignancies , yet resistance is expected on the basis of preclinical models . We recently showed that vinblastine can dramatically sensitize several leukemia cell lines to ABT-737 ( the experimental congener of DB05764 ) . The goal of these experiments was to determine the impact of vinblastine on ABT-737 sensitivity in CLL cells isolated from peripheral blood and to define the underlying mechanism . Freshly isolated CLL cells from 35 patients , as well as normal lymphocytes and platelets , were incubated with various microtubule-disrupting agents plus ABT-737 to assess sensitivity to the single agents and the combination . ABT-737 and vinblastine displayed a range of sensitivity as single agents , and vinblastine markedly sensitized all CLL samples to ABT-737 within six hours . Vinblastine potently induced the proapoptotic protein Q13794 ( Q13794 ) in both time- and dose-dependent manner and this was required for the observed apoptosis . Combretastatin A4 , which dissociates microtubules by binding to a different site , had the same effect , confirming that interaction of these agents with microtubules is the initial target . Similarly , vincristine and vinorelbine induced Q13794 and enhanced CLL sensitivity to ABT-737 . Furthermore , vinblastine plus ABT-737 overcame stroma-mediated resistance to ABT-737 alone . Apoptosis was induced with clinically achievable concentrations with no additional toxicity to normal lymphocytes or platelets . These results suggest that vinca alkaloids may improve the clinical efficacy of DB05764 in patients with CLL . P35968 -5169 , a new gastrointestinal prokinetic agent , enhances gastric contractile and emptying activities in dogs and rats . P35968 -5169 , 4-amino-5-chloro-N-[1-(3-fluoro-4-methoxybenzyl)piperidin-4-yl]-2-(2-hydroxyethoxy)benzamide hydrochloride dihydrate , is a new prokinetic with a dual action , i.e. , stimulation of the Q13639 receptor and antagonism of the dopamine D2 receptor . In this study , we determined in vitro activities of P35968 -5169 towards both receptors and demonstrated the effect of the compound on gastrointestinal motor activity in conscious dogs and rats . In dogs , intravenous P35968 -5169 stimulated upper gastrointestinal motility in the fasting state and also eliminated the depressive effect of 3,4-dihydroxyphenylalanine ( DB01235 ) on this motility in the postprandial state . The effect of P35968 -5169 on gastric emptying was further characterized by the use of three rat gastroparesis models ( dopamine D2 receptor agonist (quinpirol)- , abdominal surgery- , or combined-situation-induced ) . DB01184 ( a dopamine D2 receptor antagonist ) was effective in the quinpirol-delay and combination-delay models , and cisapride and mosapride ( Q13639 receptor agonists ) were effective in the surgery-delay model . Only P35968 -5169 eliminated the delay of gastric emptying in all three models . In addition , P35968 -5169 accelerated emptying to above the normal level in the combination-delay model . These results suggest that P35968 -5169 would be effective in various types of gastric ileus caused by different mechanisms . Improved expression of kinases in Baculovirus-infected insect cells upon addition of specific kinase inhibitors to the culture helpful for structural studies . As exemplified by three cases , we show that the addition of a small molecular weight inhibitor to the culture of Baculovirus-infected insect cells can dramatically improve the expression of a recombinant kinase . The expression of the tyrosine kinase P35968 was sevenfold higher and mainly in a soluble form , when the P35968 inhibitor DB04879 was added to the culture at the time of Baculovirus infection . The expression of the catalytic domain of the serine/threonine kinase PKCtheta , which is otherwise not possible with the Baculovirus expression system , was expressed mainly soluble at 120mg/L by the addition of the PKC inhibitor O43521 XI to the culture of Baculovirus-infected insect cells . For Abl kinase , the expression could also be significantly increased by the addition of the Abl kinase inhibitor STI571 to the culture . For all three kinases , this method had previously been applied by us for the improved production of kinase/inhibitor complex protein , leading to the co-crystal structures . It is shown here at the cases P35968 - DB04879 and PKCtheta- O43521 XI , that the stimulatory effect of an inhibitor on kinase expression is applicable under many culture conditions . The presented method represents a valuable tool to obtain structural knowledge on kinase-inhibitor complexes . 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 . Critical role of P21453 and integrin β4 in P14210 /c- DB00134 -mediated increases in vascular integrity . Vascular endothelial cell ( EC ) barrier integrity is critical to vessel homeostasis whereas barrier dysfunction is a key feature of inflammatory disorders and tumor angiogenesis . We previously reported that hepatocyte growth factor ( P14210 ) -mediated increases in EC barrier integrity are signaled through a dynamic complex present in lipid rafts involving its receptor , c- DB00134 . We extended these observations to confirm that P21453 ( sphingosine 1-phosphate receptor 1 ) and integrin β4 ( P16144 ) are essential participants in P14210 -induced EC barrier enhancement . Immunoprecipitation experiments demonstrated P14210 -mediated recruitment of c- DB00134 , P16144 and P21453 to caveolin-enriched lipid rafts in human lung EC with direct interactions of c- DB00134 with both P21453 and P16144 accompanied by c- DB00134 -dependent P21453 and P16144 transactivation . Reduced P21453 expression ( siRNA ) attenuated both P16144 and Rac1 activation as well as c- DB00134 / P16144 interaction and resulted in decreased transendothelial electrical resistance . Furthermore , reduced P16144 expression attenuated P14210 -induced c- DB00134 activation , c- DB00134 / P21453 interaction , and effected decreases in Q14703 - and P14210 -induced EC barrier enhancement . Finally , the c- DB00134 inhibitor , DB05030 , suppressed P14210 -induced c- DB00134 activation as well as P21453 and P16144 transactivation . These results support a critical role for P21453 and P16144 transactivation as rate-limiting events in the transduction of P14210 signals via a dynamic c- DB00134 complex resulting in enhanced EC barrier integrity . Molecular and behavioral effects mediated by Gs-coupled adenosine A2a , but not serotonin 5-Ht4 or 5-Ht6 receptors following antipsychotic administration . Typical antipsychotic agents are potent antagonists of Gi-coupled dopamine D2 receptors , but their mechanisms of action following this initial blockade remain poorly understood . We hypothesized that in striatal neurons , interruption of this inhibitory dopamine D2 input would unmask endogenous striatal Gs-coupled receptors . An increase in DB02527 levels generated by these unopposed receptors would then lead to the well-described behavioral and molecular effects of antipsychotic administration such as catalepsy and striatal c-fos and neurotensin gene transcription . We examined three striatal Gs-coupled receptor systems ( serotonin Q13639 , serotonin P50406 and adenosine A2a ) to assess their potential involvement in the mechanism of action of the typical antipsychotic haloperidol . Antagonists of each of these three receptor systems together with a 1 mg/kg dose of haloperidol were co-administered to Sprague-Dawley rats , and both the degree of catalepsy produced in the animals and the induction of striatal c-fos and neurotensin messenger RNAs were measured . Both the specific adenosine A2a antagonist 8-(3-chlorostyryl)-caffeine and the general adenosine antagonist theophylline reduced haloperidol-dependent induction of striatal neurotensin and c-fos messenger RNA . Administration of these agents also greatly reduced the degree of catalepsy induced by haloperidol . Antagonists of the P50406 receptor failed to block the induction of striatal messenger RNAs , but the P50406 antagonist clozapine ( an important atypical antipsychotic agent in its own right ) was a potent inhibitor of catalepsy . Q13639 agents were unable to alter haloperidol 's effects on striatal messenger RNA levels or catalepsy . We conclude that the striatal Gs-coupled adenosine A2a receptor is an important mediator of the molecular and behavioral sequelae following haloperidol administration .

MH_train_1162

MH_train_1162

MH_train_1162

interacts_with DB06209?

multiple_choice

' VASPFix ' for measurement of P50552 phosphorylation in platelets and for monitoring effects of Q9H244 antagonists . P50552 ( P50552 ) is phosphorylated and dephosphorylated consequent to increases and decreases in cyclic nucleotide levels . Monitoring changes in P50552 phosphorylation is an established method for indirect measurement of cyclic nucleotides . Here we describe the use of an innovative cocktail , VASPFix , which allows sensitive and reproducible measurement of phosphorylated P50552 ( P50552 -P ) in a simple , single-step procedure using cytometric bead technology . Frozen VASPFix-treated samples are stable for at least six months prior to analysis . We successfully used VASPFix to measure P50552 -P in platelets in both platelet-rich plasma and blood in response to compounds that increase ( dibutyryl DB02527 , adenosine , iloprost , PGE1 ) and decrease ( ADP , PGE1 ) DB02527 , and to determine the effects of certain receptor antagonists on the results obtained . The change in P50552 -P brought about by adding ADP to PGE1-stimulated platelets is a combination of the effect of ADP at the Q9H244 receptor and of PGE1 at both IP and EP3 receptors . For iloprost-stimulated platelets EP3 receptors are not involved . A procedure in which iloprost , ADP and VASPFix were used to determine effectiveness of clopidogrel and prasugrel in patients was compared with an established commercial procedure that uses PGE1 and ADP ; the latter produced higher platelet reactivity values that were the result of PGE1 interacting with platelet EP3 receptors . We conclude that VASPFix can be used both as a research tool and for clinical investigations and provides better specificity for Q9H244 receptor inhibition . The latter confers a distinct advantage over existing methods used to monitor effects of Q9H244 antagonists on platelet function . 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 . 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 . 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 . Q13639 receptor activation facilitates recovery from synaptic rundown and increases transmitter release from single varicosities of myenteric neurons . 5-HT(4) receptor agonists facilitate synaptic transmission in the enteric nervous system , and these drugs are used to treat constipation . In the present study , we investigated the effects of the 5-HT(4) receptor agonist , renzapride , on rundown and recovery of fast excitatory postsynaptic potentials ( fEPSPs ) during and after trains of stimulation and on transmitter release from individual myenteric neuronal varicosities . Intracellular electrophysiological methods were used to record fEPSPs from neurons in longitudinal muscle myenteric plexus preparations of guinea pig ileum in vitro . During trains of supramaximal electrical stimulation ( 10 Hz , 2 s ) , fEPSP amplitude declined ( time constant = 0.6 +/- 0.1 s ) from 17 +/- 2 mV to 0.7 +/- 0.3 mV . DB04917 ( 0.1 microM ) did not change the time constant for fEPSP rundown , but it decreased the time constant for recovery of fEPSP amplitude after the stimulus train from 7 +/- 2 s to 1.6 +/- 0.2 s ( P < 0.05 ) . 5-HT ( 0.1 microM ) also increased fEPSPs and facilitated recovery from rundown . The adenylate cyclase activator , forskolin ( 1 muM ) , mimicked the actions of renzapride and 5-HT , whereas H-89 , a protein kinase A ( PKA ) inhibitor , blocked the effects of renzapride . We used nicotinic acetylcholine receptor containing outside-out patches obtained from myenteric neurons maintained in primary culture to detect acetylcholine release from single varicosities . DB04917 ( 0.1 microM ) increased release probability twofold . We conclude that 5-HT(4) receptors activate the adenylyl cyclase-PKA pathway to increase acetylcholine release from single varicosities and to accelerate recovery from synaptic rundown . These responses may contribute to the prokinetic actions of 5-HT(4) receptor agonists . 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 . Q92847 agonist ( DB05657 ) accelerates gastric emptying in adults with diabetes and symptomatic gastroparesis . BACKGROUND : DB05657 is a synthetic , selective ghrelin agonist in development for gastroparesis . AIM : To assess safety and effects of DB05657 in diabetes patients with symptomatic gastroparesis . METHODS : Adults with type 1 or type 2 diabetes mellitus received placebo and DB05657 ( 80 , 160 , 320 or 600 microg/kg ) infusions in a cross-over manner following a radiolabelled meal . Blood glucose levels were stabilized using a hyperinsulinemic-euglycemic clamp . Primary endpoints were gastric half emptying and latency times . Secondary measures included assessment of gastroparesis symptoms and endocrine responses . RESULTS : Ten patients with type 1 ( n = 7 ) or 2 ( n = 3 ) diabetes , moderate-to-severe gastroparesis symptoms and > or =29 % retention 4 h after a radiolabelled solid meal were enrolled . DB05657 produced significant reductions in solid meal half-emptying ( 20 % , P = 0.043 ) and latency ( 34 % , P = 0.037 ) times vs. placebo . Reductions in overall postmeal symptom intensity ( 24 % ) and postprandial fullness ( 37 % ) following DB05657 infusion were not statistically significant . Most adverse events were mild and self-limiting and there were no identifiable differences in numbers or types of adverse events between DB05657 and placebo . CONCLUSIONS : This proof-of-concept study demonstrates that the ghrelin agonist DB05657 is well-tolerated in diabetes patients with moderate-to-severe chronic gastroparesis and shows statistically significant improvements in gastric emptying . The use of the VerifyNow Q9H244 point-of-care device to monitor platelet function across a range of Q9H244 inhibition levels following prasugrel and clopidogrel administration . Variability in response to antiplatelet agents has prompted the development of point-of-care ( POC ) technology . In this study , we compared the VerifyNow Q9H244 ( VN- Q9H244 ) POC device with light transmission aggregometry ( P01374 ) in subjects switched directly from clopidogrel to prasugrel . Healthy subjects on aspirin were administered a clopidogrel 600 mg loading dose ( LD ) followed by a 75 mg/d maintenance dose ( MD ) for 10 days . Subjects were then switched to a prasugrel 60 mg LD and then 10 mg/d MD for 10 days ( n = 16 ) , or to a prasugrel 10 mg/d MD for 11 days ( n = 19 ) . Platelet function was measured by P01374 and VN- Q9H244 at baseline and after dosing . DB00758 600 mg LD/75 mg MD treatment led to a reduction in P2Y(12) reaction units ( PRU ) from baseline . A switch from clopidogrel MD to prasugrel 60 mg LD/10 mg MD produced an immediate decrease in PRU , while a switch to prasugrel 10 mg MD resulted in a more gradual decline . Consistent with the reduction in PRU , device-reported percent inhibition increased during both clopidogrel and prasugrel regimens . Inhibition of platelet aggregation as measured by P01374 showed a very similar pattern to that found with VN- Q9H244 measurement , irrespective of treatment regimens . The dynamic range of VN- Q9H244 appeared to be narrower than that of P01374 . With two different thienopyridines , the VN- Q9H244 device , within a somewhat more limited range , reflected the overall magnitude of change in aggregation response determined by P01374 . The determination of the clinical utility of such POC devices will require their use in clinical outcome studies . 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 . Involvement of reactive oxygen species in O00206 -dependent activation of NF-kappa B . Although oxidative stress has been thought to play a general role in the activation of NF-kappaB , the involvement of reactive oxygen species ( ROS ) in facilitating nuclear translocation of NF-kappaB in neutrophils has not been described . In addition , the mechanisms by which ROS modulate the transcriptional activity of NF-kappaB in response to O00206 ( O00206 ) -dependent signaling are not well characterized . To examine these issues , oxidant-dependent signaling events downstream of O00206 were investigated in neutrophils stimulated with LPS . Pretreatment of neutrophils with the antioxidants DB06151 or DB00163 prevented LPS-induced nuclear translocation of NF-kappaB . Antioxidant treatment of LPS-stimulated neutrophils also inhibited the production of proinflammatory cytokines ( P01375 , macrophage inflammatory protein-2 , and IL-1beta ) , as well as activation of the kinases O15111 alpha , O15111 beta , p38 , Akt , and extracellular receptor-activated kinases 1 and 2 . The decrease in cytoplasmic levels of P25963 produced by exposure of neutrophils to LPS was prevented by DB06151 or DB00163 . Activation of IL-1R-associated kinase-1 ( P51617 ) and Q9NWZ3 in response to LPS stimulation was inhibited by antioxidants . These results demonstrate that proximal events in O00206 signaling , at or antecedent to P51617 and Q9NWZ3 activation , are oxidant dependent and indicate that ROS can modulate NF-kappaB-dependent transcription through their involvement in early O00206 -mediated cellular responses . 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 . DB04998 inhibits activation of nuclear factor-kappaB ( NF-kappaB ) by forming a complex with NF-kappaB essential modulator ( Q9Y6K9 ) and nucleolin . DB04998 , also known as DB04998 , is an experimental anticancer drug that recently entered human clinical trials . It is a member of a novel class of antiproliferative agents known as G-rich oligonucleotides ( P09341 ) , which are non-antisense , guanosine-rich phosphodiester oligodeoxynucleotides that form stable G-quadruplex structures . The biological activity of GROs results from their binding to specific cellular proteins as aptamers . One important target protein of GROs has been previously identified as nucleolin , a multifunctional protein expressed at high levels by cancer cells . Here , we report that DB04998 also associates with nuclear factor-kappaB ( NF-kappaB ) essential modulator ( Q9Y6K9 ) , which is a regulatory subunit of the inhibitor of kappaB ( IkappaB ) kinase ( IKK ) complex , and also called IKKgamma . In the classic NF-kappaB pathway , the IKK complex is required for phosphorylation of P25963 and subsequent activation of the transcription factor NF-kappaB . We found that treatment of cancer cells with DB04998 inhibits IKK activity and reduces phosphorylation of P25963 in response to tumor necrosis factor-alpha stimulation . Using a reporter gene assay , we showed that DB04998 blocks both tumor necrosis factor-alpha-induced and constitutive NF-kappaB activity in human cancer cell lines derived from cervical , prostate , breast , and lung carcinomas . In addition , we showed that , in DB04998 -treated cancer cells , Q9Y6K9 is coprecipitated by nucleolin , indicating that both proteins are present in the same complex . Our studies suggest that abrogation of NF-kappaB activity may contribute to the anticancer effects of DB04998 and that nucleolin may play a previously unknown role in regulating the NF-kappaB pathway . Distribution of polyamines and their biosynthetic enzymes in intestinal adaptation . P11926 ( ODC ) and the polyamines have been shown to be important for growth processes in the intestinal mucosa . The highest activity of ODC is found in the differentiated , nonproliferating villus-tip cells rather than in the rapidly proliferating undifferentiated crypt cells . During poststarvation refeeding and lactation , we now show that increases in ODC activity paralleled the time course of mucosal hyperplasia and thymidine incorporation . Increases in ODC ( threefold ) were similar in villus and crypt cells , and the villus-crypt gradient of decreasing ODC activity ( 40:1 ) was maintained . The activity of the other polyamine biosynthetic enzyme , S-adenosylmethionine decarboxylase ( P18827 ) , was highest in the crypt cells in the basal state and increased throughout the entire villus-crypt axis during refeeding and lactation , preserving a villus-crypt gradient opposite to that of ODC . During hyperplasia , all three polyamines increased . DB01917 was highest in the villus-tip cells , paralleling ODC activity , whereas spermidine and spermine were highest in the crypt cells and paralleled the distribution of P18827 activity . Thus P18827 activity and spermidine and spermine content may play a more important role than ODC and putrescine in regulation of intestinal mucosal proliferation . It is also possible that the threefold increases in the low levels of ODC in the crypt cells are adequate to trigger cell proliferation , whereas the higher ODC levels in villus cells may represent an association with the differentiation of the enterocytes . A multi-analyte assay for the non-invasive detection of bladder cancer . Accurate urinary assays for bladder cancer ( BCa ) detection would benefit both patients and healthcare systems . Through genomic and proteomic profiling of urine components , we have previously identified a panel of biomarkers that can outperform current urine-based biomarkers for the non-invasive detection of BCa . Herein , we report the diagnostic utility of various multivariate combinations of these biomarkers . We performed a case-controlled validation study in which voided urines from 127 patients ( 64 tumor bearing subjects ) were analyzed . The urinary concentrations of 14 biomarkers ( P10145 , P14780 , P09238 , P18827 , P55774 , P05121 , P16070 , P15692 , P03950 , Q16790 , A1AT , P10451 , PTX3 , and P02649 ) were assessed by enzyme-linked immunosorbent assay ( ELISA ) . Diagnostic performance of each biomarker and multivariate models were compared using receiver operating characteristic curves and the chi-square test . An 8-biomarker model achieved the most accurate BCa diagnosis ( sensitivity 92 % , specificity 97 % ) , but a combination of 3 of the 8 biomarkers ( P10145 , P15692 , and P02649 ) was also highly accurate ( sensitivity 90 % , specificity 97 % ) . For comparison , the commercial BTA-Trak ELISA test achieved a sensitivity of 79 % and a specificity of 83 % , and voided urine cytology detected only 33 % of BCa cases in the same cohort . These data show that a multivariate urine-based assay can markedly improve the accuracy of non-invasive BCa detection . Further validation studies are under way to investigate the clinical utility of this panel of biomarkers for BCa diagnosis and disease monitoring . 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 . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS ) Role of NF-κB activation in LPS-induced endothelial barrier breakdown . Endothelial barrier breakdown contributes to organ failure in sepsis . The key mechanism by which the potent sepsis inductor lipopolysaccharide ( LPS ) disrupts the endothelial barrier is controversial . Here , we tested the hypothesis that NF-κB activation is critically involved in endothelial barrier breakdown . Application of LPS to monolayers of porcine pulmonary artery endothelial cells ( PAEC ) and human dermal microvascular endothelial cells ( HDMEC ) induced a rapid and sustained activation of NF-κB as revealed by translocation of its subunit p65 into the nuclei in nuclear extraction assays and by immunostaining . Measurements of transendothelial electrical resistance ( Q9NZ01 ) and intercellular gap formation demonstrated significant breakdown of endothelial barrier properties following LPS treatment for 3 h . Interestingly , monolayers recovered spontaneously beginning after 10 h . Increased DB02527 prevented LPS-induced loss of endothelial barrier properties , but did not block NF-κB activation . Application of the cell-permeable Q9Y6K9 -binding domain ( NBD ) synthetic peptide was effective to prevent NF-κB activation , but did neither block LPS-induced loss of Q9NZ01 nor intercellular gap formation . NBD peptide alone did not alter endothelial barrier properties , but enhanced the barrier-compromising effects when applied in combination with LPS . Similarly , siRNA-mediated knock-down of p65 in HDMECs did not prevent LPS-induced barrier breakdown . Known targets of NF-κB-derived protein expression of caveolin or vasodilator-stimulated phosphoprotein ( P50552 ) remained unaltered by LPS treatment of endothelial cells . In summary , our data indicate that NF-κB activation by LPS is not critically involved in disruption of endothelial barrier properties . Rather , our data suggest that NF-κB activation acts as a part of a rescue mechanism . Ghrelin protects against renal damages induced by angiotensin-II via an antioxidative stress mechanism in mice . We explored the renal protective effects by a gut peptide , Ghrelin . Daily peritoneal injection with Ghrelin ameliorated renal damages in continuously angiotensin II ( AngII ) -infused C57BL/6 mice as assessed by urinary excretion of protein and renal tubular markers . AngII-induced increase in reactive oxygen species ( ROS ) levels and senescent changes were attenuated by Ghrelin . Ghrelin also inhibited AngII-induced upregulations of transforming growth factor-β ( TGF-β ) and plasminogen activator inhibitor-1 ( P05121 ) , ameliorating renal fibrotic changes . These effects were accompanied by concomitant increase in mitochondria uncoupling protein , P55851 as well as in a key regulator of mitochondria biosynthesis , PGC1α . In renal proximal cell line , HK-2 cells , Ghrelin reduced mitochondria membrane potential and mitochondria-derived ROS . The transfection of P55851 siRNA abolished the decrease in mitochondria-derived ROS by Ghrelin . Ghrelin ameliorated AngII-induced renal tubular cell senescent changes and AngII-induced TGF-β and P05121 expressions . Finally , Q92847 , growth hormone secretagogue receptor ( Q92847 ) -null mice exhibited an increase in tubular damages , renal ROS levels , renal senescent changes and fibrosis complicated with renal dysfunction . Q92847 -null mice harbored elongated mitochondria in the proximal tubules . In conclusion , Ghrelin suppressed AngII-induced renal damages through its P55851 dependent anti-oxidative stress effect and mitochondria maintenance . Ghrelin/ Q92847 pathway played an important role in the maintenance of ROS levels in the kidney . Risks of human limb deficiency anomalies associated with 29 SNPs of genes involved in homocysteine metabolism , coagulation , cell-cell interactions , inflammatory response , and blood pressure regulation . This study explored risks of limb deficiency anomalies associated with 29 single nucleotide polymorphisms ( SNPs ) of genes involved in homocysteine metabolism , coagulation , cell-cell interaction , inflammatory response , and blood pressure regulation . The authors genotyped 96 cases and 437 non-malformed controls from a California population-based case-control study ( 1987-1988 birth cohort ) . Increased risk of limb anomaly was observed for three SNPs : heterozygosity for P12259 Arg506Gln , with an odds ratio ( OR ) of 2.5 ( 95 % confidence interval ( CI ) , 1.0 , 6.5 ) ; heterozygosity for P01375 (-376)G > A , OR 2.1 ( 0.7 , 6.2 ) ; and homozygosity for P01160 2238T > C , OR 4.0 ( 1.1 , 15.4 ) . We hypothesized that effects of variant genotypes in the presence of maternal smoking , and/or in the absence of supplement intake , may exceed effects of any of these factors alone . In particular , findings for polymorphisms in P05121 , P17301 , P16581 , P01375 , P01374 , P01160 , P16520 , and P07550 supported the hypotheses , both for smoking and for supplement intake . These results suggest involvement of genetic variation of biologically relevant candidate genes , and gene-environment interaction , for some limb anomalies whose pathogenesis may be related to altered vascular tone or integrity . Internet resources in GPCR modelling . G-Protein coupled receptors ( GPCRs ) , one of the most important families of drug targets , belong to the super family of integral membrane proteins characterized by seven transmembrane helices . Because they are difficult to crystallize , the three dimensional structure of these receptors have not yet been determined by X-ray crystallography , except one . In the absence of a 3-D structure , in-silico approaches for solving the structure of this class of proteins are widely used and provide valuable information for structure based drug design . There are several web servers and computer programs available that automate the modelling process of GPCRs . Some of these include Modeller , Swiss-Model server , Homer , etc . Using these tools reliable homology models of human histamine H1 receptor ( P35367 ) and thrombin receptor ( P25116 ) have been generated which explain the binding mode of the standard antagonists of these receptors and may be useful in designing their novel antagonists . Effect of licofelone -- a dual P36551 /5- P28300 inhibitor in intracerebroventricular streptozotocin-induced behavioral and biochemical abnormalities in rats . The present study was designed to investigate the effect of licofelone-a dual cyclooxygenase/ P09917 ( P36551 /5- P28300 ) inhibitor in intracerebroventricular streptozotocin ( ICV- Q11206 ) -induced cognitive deficit and biochemical abnormalities in rats . ICV- Q11206 is a widely used model of sporadic Alzheimer 's disease . In this study , Q11206 was administered intracerebroventricular (i.c.v.)-bilaterally 3 mg/kg in rats . The Q11206 -injected rats were treated with different doses of licofelone ( 2.5 , 5 , and 10 mg/kg , p.o. ) for 21 days . Cognitive functions were assessed by using Morris water maze and passive avoidance task . Levels of malondialdehyde ( MDA ) , nitrite , reduced glutathione ( DB00143 ) , and acetylcholinesterase ( P22303 ) activity were determined to check oxidative stress and cholinergic function . Cytokine levels ( IL-1β and P01375 -α ) were also determined as markers of neuroinflammation . Administration of Q11206 caused a significant increase in P22303 activity and cognitive dysfunction . Increased oxidative stress and the proinflammatory cytokine levels were also observed following Q11206 administration in rats . DB04725 treatment attenuated Q11206 -induced cholinergic hypofunction and cognitive deficit in rats . In addition , licofelone attenuated Q11206 -induced oxidative stress and elevated cytokine levels . The cognitive enhancement following licofelone administration in Q11206 rats may be due to its ability to restore cholinergic functions or its antioxidant activity . These observed results suggest the therapeutic potential of dual P36551 /5- P28300 inhibitors in neurodegenerative disorders associated with oxidative stress and cognitive impairment . Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice . Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined . The immediate phase reaction ( IPR ) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined . The inhibition of IPR by cetirizine and mequitazine were potent , but those by cyproheptadine and diphenhydramine were weak . The later phase reaction ( LPR ) assessed at 24 hours after antigen application was inhibited by chlorpheniramine , oxatomide , ketotifen , mequitazine , emedastine , terfenadine and azelastine . The inhibition of LPR by emedastine was potent , but those by ketotifen and terfenadine were only partial . DB01084 inhibited both IPR and LPR comparably . Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction , and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR . P35367 antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism , which may have an additional benefit for the treatment of allergic diseases . Prasugrel is effective and safe for neurointerventional procedures . BACKGROUND : DB00758 bisulfate and aspirin are routinely administered as dual antiplatelet agents for many neurointerventional procedures , especially for intravascular stent placement . Many patients are non-responsive to clopidogrel , either secondary to drug interactions or from variations of cytochrome P450 enzymes . Prasugrel ( brand name DB06209 , Eli Lilly and Company , Indianapolis , IN , USA ) is a new antiplatelet agent that has been utilized extensively in patients undergoing cardiovascular procedures but its safety and efficacy during neurointerventional procedures have not been evaluated . OBJECTIVE : To examine whether prasugrel is a safe and effective alternative to clopidogrel for neurointerventional procedures , especially in those patients who are either non-responders or allergic to clopidogrel . METHODS : The medical records of all patients undergoing neurointerventional procedures at our institution who received prasugrel between January 2009 and July 2011 were retrospectively reviewed . A systematic chart review was performed and the following data were recorded : demographics , aneurysm location , endovascular techniques , peri- and post-procedural complications , hemorrhagic complications , clinical outcome and angiographic outcome . RESULTS : 16 patients undergoing neurointerventional procedures received prasugrel over a 2 year interval . All patients who had follow-up studies of Q9H244 inhibition had immediate therapeutic response to prasugrel . There were no complications related to ischemic or intracranial hemorrhage . CONCLUSION : Prasugrel is a viable alternative to clopidogrel for patients undergoing neurointerventional procedures who are non-responders to clopidogrel . Further study is needed to evaluate the safety , efficacy and cost-effectiveness of prasugrel compared with clopidogrel for patients undergoing neurointerventional procedures . 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 .

MH_train_1163

MH_train_1163

MH_train_1163

interacts_with DB00991?

multiple_choice

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 . 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 . A calmodulin-binding site on cyclin E mediates Ca2+-sensitive P55008 /s transitions in vascular smooth muscle cells . DB01373 transients are known to control several transition points in the eukaryotic cell cycle . For example , we have previously shown that a coordinate elevation in the intracellular free calcium ion concentration is required for P55008 - to S-phase cell cycle progression in vascular smooth muscle cells ( VSMC ) . However , the molecular basis for this Ca2+ sensitivity was not known . Using buffers with differing [ Ca2+ ] , we found that the kinase activity of mouse and human cyclin E/ P24941 , but not other P55008 /S-associated cell cycle complexes , was responsive to physiological changes in [ Ca2+ ] . We next determined that this Ca2+-responsive kinase activity was dependent on a direct interaction between calmodulin ( P62158 ) , one of the major Ca2+-signal transducers of eukaryotic cells , and cyclin E. Pharmacological inhibition of P62158 abrogated the Ca2+ sensitivity of cyclin E/ P24941 and retarded mouse VSMC proliferation by causing P55008 arrest . We next defined the presence of a highly conserved 22 amino acid N-terminal P62158 -binding motif in mammalian cyclin E genes ( dissociation constant , 1.5+/-0.1 micromol/L ) and showed its essential role in mediating Ca2+-sensitive kinase activity of cyclin E/ P24941 . Mutant human cyclin E protein , lacking this P62158 -binding motif , was incapable of binding P62158 or responding to [ Ca2+ ] . Taken together , these findings reveal P62158 -dependent cyclin E/ P24941 activity as a mediator of the known Ca2+ sensitivity of the P55008 /S transition of VSMC . Identification of a de novo thymidylate biosynthesis pathway in mammalian mitochondria . The de novo and salvage dTTP pathways are essential for maintaining cellular dTTP pools to ensure the faithful replication of both mitochondrial and nuclear DNA . Disregulation of dTTP pools results in mitochondrial dysfunction and nuclear genome instability due to an increase in uracil misincorporation . In this study , we identified a de novo dTMP synthesis pathway in mammalian mitochondria . Mitochondria purified from wild-type Chinese hamster ovary ( CHO ) cells and HepG2 cells converted DB03800 to dTMP in the presence of NADPH and serine , through the activities of mitochondrial serine hydroxymethyltransferase ( P34897 ) , thymidylate synthase ( P04818 ) , and a novel human mitochondrial dihydrofolate reductase ( P00374 ) previously thought to be a pseudogene known as dihydrofolate reductase-like protein 1 ( Q86XF0 ) . Human Q86XF0 , P34897 , and P04818 were localized to mitochondrial matrix and inner membrane , confirming the presence of this pathway in mitochondria . Knockdown of Q86XF0 using siRNA eliminated P00374 activity in mitochondria . Q86XF0 expression in CHO glyC , a previously uncharacterized mutant glycine auxotrophic cell line , rescued the glycine auxotrophy . De novo thymidylate synthesis activity was diminished in mitochondria isolated from glyA CHO cells that lack P34897 activity , as well as mitochondria isolated from wild-type CHO cells treated with methotrexate , a P00374 inhibitor . De novo thymidylate synthesis in mitochondria prevents uracil accumulation in mitochondrial DNA ( mtDNA ) , as uracil levels in mtDNA isolated from glyA CHO cells was 40 % higher than observed in mtDNA isolated from wild-type CHO cells . These data indicate that unlike other nucleotides , de novo dTMP synthesis occurs within mitochondria and is essential for mtDNA integrity . P08246 inhibitors as treatment for P48444 . Chronic obstructive pulmonary disease , characterised by a slowly progressive , irreversible airways limitation , is a major worldwide cause of chronic morbidity and mortality . The imbalance between human neutrophil elastase and endogenous antiproteases may cause excess human neutrophil elastase in pulmonary tissues , which may be considered a major pathogenic factor in chronic obstructive pulmonary disease . Great effort has been devoted to finding a method to restore the balance , resulting in the discovery of potent two-typed small-molecular-weight human neutrophil elastase inhibitors . In the application of chronic obstructive pulmonary disease therapy , the human neutrophil elastase inhibitors mainly focused upon include ONO-5046 , MR-889 , L-694,458 , CE-1037 , GW-311616 and TEI-8362 as the acyl-enzyme inhibitors ; and DB03925 , AE-3763 , FK-706 , ICI-200,880 , ZD-0892 and ZD-8321 as the transition-state inhibitors . In this review , various problems that remain to be solved in the clinical use of human neutrophil elastase inhibitors are discussed . Inhibition of cyclooxygenases 1 and 2 by the phospholipase-blocker , arachidonyl trifluoromethyl ketone . BACKGROUND AND PURPOSE : Arachidonyl trifluoromethyl ketone ( Q06187 ) is widely used as an inhibitor of cytosolic group IV phospholipase A(2) ( cPLA(2) ) and calcium-independent group VI phospholipase A(2) ( iPLA(2) ) . Q06187 thus reduces arachidonic acid ( AA ) substrate for cyclooxygenase ( P36551 ; also known as prostaglandin H synthase ) and attenuates prostaglandin ( PG ) synthesis . It has been shown previously , that Q06187 blocks thromboxane B(2) production induced by exogenous AA in human platelets . It remains , however , unknown whether Q06187 also directly modulates the activity of cyclooxygenase ( P36551 ) . EXPERIMENTAL APPROACH : Time courses for inhibition of P36551 by Q06187 was obtained using osteoblast-like MC3T3-E1 cells , with exogenous AA as substrate and the pure enzymes P23219 and P35354 . PGE(2) was measured by GC-MS . KEY RESULTS : Q06187 was a potent inhibitor of P23219 and P35354 with IC(50) values of 0.5 and 0.1 microM in MC3T3-E1 cells and of 1.7 and 2.6 microM using the pure enzymes . Inhibition was reversible , with slow- and tight-binding characteristics . The arachidonyl carbon chain was essential , as the saturated palmitoyl analogue had no effect . CONCLUSIONS AND IMPLICATIONS : Attenuation of PG synthesis by Q06187 is taken to be the consequence of PLA(2) inhibition and the findings of many studies are interpreted on that basis . If there are , however , alternative routes for AA liberation ( such as phospholipase C/diacyl glycerol lipase or phospholipase D ) , this interpretation can lead to false conclusions . As Q06187 is a widely used and important pharmacological tool in eicosanoid research , knowledge of its interactions with other major enzymes of the cascade is of considerable importance . In vitro effects of gonadotropin-releasing hormone ( DB00644 ) on Leydig cells of adult alpaca ( Lama pacos ) testis : P30968 immunolocalization , testosterone and prostaglandin synthesis , and cyclooxygenase activities . The main objective of this study was to examine the modulatory in vitro effects of gonadotropin-releasing hormone ( DB00644 ) on isolated Leydig cells of adult alpaca ( Lama pacos ) testis . We first evaluated the presence of P30968 ( GnRHR ) and cyclooxygenase ( P36551 ) 1 and P35354 in alpaca testis . We then studied the in vitro effects of buserelin ( DB00644 analogue ) , antide ( DB00644 antagonist ) , and buserelin plus antide or inhibitor of phospholipase C ( compound 48/80 ) and COXs ( acetylsalicylic acid ) on the production of testosterone , PGE(2) , and P49763 (2α) and on the enzymatic activities of P23219 and P35354 . Immunoreactivity for GnRHR was detected in the cytoplasm of Leydig cells and in the acrosomal region of spermatids . P23219 and P35354 immunosignals were noted in the cytoplasm of spermatogonia , spermatocytes , spermatids , Leydig cells , and Sertoli cells . Western blot analysis confirmed the GnRHR and P23219 presence in alpaca testis . The in vitro experiments showed that buserelin alone increased ( P < 0.01 ) and antide and buserelin plus acetylsalicylic acid decreased ( P < 0.01 ) testosterone and P49763 (2α) production and P23219 activity , whereas antide and compound 48/80 counteracted buserelin effects . Prostaglandin E(2) production and P35354 activity were not affected by buserelin or antide . These data suggest that DB00644 directly up-regulates testosterone production in Leydig cells of adult alpaca testis with a postreceptorial mechanism that involves P98160 , P23219 , and P49763 (2α) . DB03758 activates heat shock protein expression and cardioprotection in neonatal rat cardiomyocytes . Heat shock proteins ( HSPs ) constitute an endogenous cellular defense mechanism against environmental stresses . In the past few years , studies have shown that overexpression of HSPs can protect cardiac myocytes against ischemia-reperfusion injury . In an attempt to increase the HSPs in cardiac tissue , we used the compound radicicol that activates HSP expression by binding to the P08238 kDa ( HSP90 ) . HSP90 is the main component of the cytosolic molecular chaperone complex , which has been implicated in the regulation of the heat shock factor 1 ( Q00613 ) . Q00613 is responsible for the transcriptional activation of the heat shock genes . In the present study , we show that radicicol induces HSP expression in neonatal rat cardiomyocytes , and this increase in HSPs confers cardioprotection to these cardiomyocytes . We also show that radicicol induction of the HSP and cardioprotection is dependent on the inhibition of HSP90 in cardiomyocytes . These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection . Therefore , compounds , such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents . Histaminergic ligands injected into the nucleus basalis magnocellularis differentially affect fear conditioning consolidation . The role of the nucleus basalis magnocellularis ( NBM ) in fear conditioning encoding is well established . In the present report , we investigate the involvement of the NBM histaminergic system in consolidating fear memories . The NBM was injected bilaterally with ligands of histaminergic receptors immediately after contextual fear conditioning . Histaminergic compounds , either alone or in combination , were stereotaxically administered to different groups of adult male Wistar rats and memory was assessed as conditioned freezing duration 72 h after administration . This protocol prevents interference with NBM function during either acquisition or retrieval phases , hence restricting the effect of pharmacological manipulations to fear memory consolidation . The results presented here demonstrate that post-training H3 receptors ( Q9Y5N1 ) blockade with the antagonist/inverse agonist thioperamide or activation with immepip in the NBM potentiates or decreases , respectively , freezing response at retrieval . Thioperamide induced memory enhancement seems to depend on P25021 , but not P35367 activation , as the P25021 antagonist zolantidine blocked the effect of thioperamide , whereas the P35367 antagonist DB06691 was ineffective . Furthermore , the P25021 agonist ampthamine improved fear memory expression independently of the Q9Y5N1 agonist effect . Our results indicate that activation of post-synaptic P25021 within the NBM by endogenous histamine is responsible for the potentiated expression of fear responses . The results are discussed in terms of activation of H3 auto- and heteroreceptors within the NBM and the differential effect of Q9Y5N1 ligands on fear memory consolidation in distinct brain regions . 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 . Regulation of cyclooxygenase-2 expression by heat : a novel aspect of heat shock factor 1 function in human cells . The heat-shock response , a fundamental defense mechanism against proteotoxic stress , is regulated by a family of heat-shock transcription factors ( HSF ) . In humans Q00613 is considered the central regulator of heat-induced transcriptional responses . The main targets for Q00613 are specific promoter elements ( HSE ) located upstream of heat-shock genes encoding cytoprotective heat-shock proteins ( HSP ) with chaperone function . In addition to its cytoprotective function , Q00613 was recently hypothesized to play a more complex role , regulating the expression of non-HSP genes ; however , the non-canonical role of Q00613 is still poorly understood . Herein we report that heat-stress promotes the expression of cyclooxygenase-2 ( P35354 ) , a key regulator of inflammation controlling prostanoid and thromboxane synthesis , resulting in the production of high levels of prostaglandin-E(2) in human cells . We show that heat-induced P35354 expression is regulated at the transcriptional level via Q00613 -mediated signaling and identify , by in-vitro reporter gene activity assay and deletion-mutant constructs analysis , the P35354 heat-responsive promoter region and a new distal cis-acting HSE located at position -2495 from the transcription start site . As shown by ChIP analysis , Q00613 is recruited to the P35354 promoter rapidly after heat treatment ; by using shRNA-mediated Q00613 suppression and HSE-deletion from the P35354 promoter , we demonstrate that Q00613 plays a central role in the transcriptional control of P35354 by heat . Finally , P35354 transcription is also induced at febrile temperatures in endothelial cells , suggesting that Q00613 -dependent P35354 expression could contribute to increasing blood prostaglandin levels during fever . The results identify P35354 as a human non-classical heat-responsive gene , unveiling a new aspect of Q00613 function . 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 . Structure-function studies of linear and cyclized peptide antagonists of the P30968 . Structurally new analogs of the peptidic P30968 antagonist DB00050 as well as conformationally constrained cyclized deca- or pentapeptides were synthesized and selected peptides evaluated comprehensively . To understand how structural variations of the antagonistic peptide effect pharmacodynamic properties , binding affinities and antagonistic potencies toward the human and rat P30968 were determined . Whereas large substituents in position 6 of linear peptides are compatible with high binding affinity ( K(D) < 0.5 nM ) , all cyclized peptides except the cyclo[3-10] analog D-52391 depicted low binding affinity ( K(D) > 10 nM ) . Binding affinity and antagonistic potency in vitro correlated for all peptides and surprisingly no discrimination between human and rat receptor proteins was observed . Since receptor residues W(101) and N(102) are involved in agonist and antagonist binding , equally potent but structurally different antagonists were tested for binding to the respective W(101)A and N(102)A mutants . In contrast to linear decapeptides , residues N(102) and W(101) are not involved in binding of D-23938 and W(101) is the critical residue for D-52391 binding . We conclude that although equally potent , peptidic P30968 antagonists do have distinct interactions within the ligand binding pocket . Finally , selected antagonists were tested for testosterone suppression in male rats . The duration of testosterone suppression below castration levels differed largely from 1 day for DB06785 to 27 days for D-23487 . Systemic availability became evident as the most important parameter for in vivo efficacy . DB09053 -naïve chronic lymphocytic leukemia lacks Q06187 mutations associated with treatment resistance . Inhibition of tumor necrosis factor signal transduction in endothelial cells by dimethylaminopurine . P01375 ( P01375 ) promotes diverse responses in endothelial cells that are important to the host response to infections and malignancies ; however , less is known of the postreceptor events important to P01375 action in endothelial cells than in many other cell types . Since phosphorylation cascades are implicated in cytokine signaling , the effects of the protein kinase inhibitor dimethylaminopurine ( DMAP ) on P01375 action in bovine aortic endothelial cells ( BAEC ) were investigated . In BAEC , P01375 promotes phosphorylation of eukaryotic initiation factor 4E ( P06730 ) , c-Jun N-terminal kinase ( JNK ) and ceramide-activated protein kinase activities , Jun-b expression , prostacyclin production , and , when protein synthesis is inhibited , cytotoxicity . DMAP abrogated or significantly attenuated each of these responses to P01375 , without affecting the specific binding of P01375 to its receptors . DB11320 , another agent active in the endothelium , promotes phosphorylation of elongation factor-2 ( P13639 ) and prostacyclin production , but not phosphorylation of P06730 in BAEC . DB11320 -stimulated P13639 phosphorylation was not inhibited and prostacyclin production was unaffected by DMAP . These observations demonstrate that a distinct signal transduction cascade , which can be selectively inhibited by DMAP , promotes the response of BAEC to P01375 . Thus , we have identified a reagent , DMAP , that may be useful for characterizing the P01375 signal transduction pathway . All-trans retinoic acid up-regulates Prostaglandin-E Synthase expression in human macrophages . All-trans retinoic acid ( DB00755 ) is a potent retinoid , which has been used successfully in different clinical settings as a potential drug to treat P48444 and emphysema . In the present study , we analyzed genes modulated by DB00755 by performing mRNA expression array analysis on alveolar macrophages after treatment with DB00755 . Here we observed a 375-fold up-regulation of Prostaglandin-E Synthase ( microsomal O14684 -1 , NM_004878 O14684 ) which mediates the conversion of prostaglandin H(2) ( PGH(2) ) to Prostaglandin E(2) ( PGE(2) ) . We furthermore studied the expression of O14684 after treatment with DB00755 in human monocyte-derived macrophages ( MDMs ) and bronchoalveolar lavage ( BAL ) cells . DB00755 up-regulated O14684 mRNA expression in MDMs generated with P09603 by 2500-fold whereas in P09603 + P35225 macrophages the up-regulation was only 20-fold . Similarly , DB00755 up-regulated O14684 mRNA expression by factor 1524 in BAL cells . The up-regulation of O14684 mRNA expression by DB00755 is both time and dose dependent . P35225 suppressed the DB00755 induced O14684 expression at both mRNA and protein level in MDM and BAL cells . We also observed that LPS acts synergistically with DB00755 in MDMs and strongly induces O14684 expression . DB00755 had little impact on cyclooxygenase-1 and -2 ( P23219 and -2 ) expression as compared to O14684 expression under the same experimental conditions . Furthermore , we observed an induction of PGE(2) levels by DB00755 in BAL cells . These data indicate that DB00755 is a potent inducer of O14684 expression in human macrophages but not in alternatively activated macrophages and suggest that the eicosanoid pathway is important for DB00755 action in macrophages . Modification of the reactivity of three amino-acid residues in elongation factor 2 during its binding to ribosomes and translocation . The accessibility of three amino acids of P13639 , located within highly conserved regions near the N- and C-terminal extremities of the molecule ( the E region and the DB02059 region , respectively ) to modifying enzymes has been compared within nucleotide-complexed P13639 and ribosomal complexes that mimic the pre- and posttranslocational ones : the high-affinity complex ( P13639 ) -nonhydrolysable GTP analog GuoPP[CH2]P ribosome and the low-affinity ( P13639 ) -GDP-ribosome complex , P13639 and ribosomes being from rat liver . We studied the reactivity of two highly conserved residues diphthamide-715 and DB00125 -66 , to diphtheria-toxin-dependent ADP-ribosylation and trypsin attack , and of a threonine that probably lies between residues 51 and 60 , to phosphorylation by a Ca2+/calmodulin-dependent protein kinase . DB03223 715 and this threonine residue were unreactive within the high-affinity complex but seemed fully reactive in the low-affinity complex . DB00125 -66 was resistant to trypsin in both complexes . The possible involvement of the E and DB02059 regions of P13639 in the interaction with ribosome in the two complexes is discussed . Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5-methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 -dependent enzyme methionine synthase . Q99707 specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells . [ A genetic background of ulcer diseases induced by NSAID/aspirin ] . The association between peptic ulcer diseases and polymorphisms in various genes , including P25021 , P23219 , Q16552 . Q96PD4 , MIF and Nrf2 genes , are seen . P23219 has traditionally been regarded as a constitutively expressed enzyme that generates prostaglandins for gastrointestinal integrity . The effects of NSAID/aspirin on the gastric mucosal damage are caused by the inhibition of this enzyme . A T-1676C polymorphism ( rs1330344 ) was significantly associated with the development of peptic ulcer , especially gastric ulcer . In addition , rs1330344 was also significantly associated with the development of NSAID/aspirin-induced ulcer diseases . In conclusions , the assessment for genotype of P23219 gene promoter polymorphism , especially rs1330344 , may be useful for detecting the high risk group of developing NSAID/aspirin-induced ulcer diseases . 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 .

MH_train_1164

MH_train_1164

MH_train_1164

interacts_with DB00486?

multiple_choice

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 . Passive smoke effects on cough and airways in young guinea pigs : role of brainstem DB05875 . Children raised with extended exposure to environmental tobacco smoke ( ETS ) experience increased cough and wheeze . This study was designed to determine whether extended ETS exposure enhances citric acid-induced cough and bronchoconstriction in young guinea pigs via a neurokinin-1 ( NK-1 ) receptor mechanism at the first central synapse of lung afferent neurons , the nucleus tractus solitarius . Guinea pigs were exposed to ETS from 1 to 6 weeks of age . At 5 weeks of age , guide cannulae were implanted bilaterally in the medial nucleus tractus solitarius at a site that produced apnea in response to the glutamate agonist D,L-homocysteic acid . At 6 weeks of age , either vehicle or a P25103 antagonist , DB05790 , was injected into the nucleus tractus solitarius of the conscious guinea pigs who were then exposed to citric acid aerosol . ETS exposure significantly enhanced citric acid-induced cough by 56 % and maximal Penh ( a measure of airway obstruction ) by 43 % , effects that were attenuated by the P25103 antagonist in the nucleus tractus solitarius . We conclude that in young guinea pigs extended exposure to ETS increases citric acid-induced cough and bronchoconstriction in part by an P25103 mechanism in the nucleus tractus solitarius . Ca2+ response of rat mesangial cells to DB00171 analogues . The aim of this investigation was to characterise the effects of DB00171 analogues and UTP on the single cell intracellular Ca2+ concentration ( [Ca2+]i ) in cultured rat mesangial cells . Typically , there were two phases in the Ca2+ response to the agonists , an initial fast transient peak and a subsequent slower decline , or plateau , phase . For the peak amplitude in [Ca2+]i the agonists had about equal effect . But when taking in consideration the percentage of responding cells and the integrated Ca2+ response over 1 min , the order of efficacy of nucleotide agonists ( 100 microM ) was UTP = DB00171 > ATPgammaS > ADP = 2MeS- DB00171 ( 2-methylthio- DB00171 ) . DB00640 , AMP and beta,gamma-Me- DB00171 ( 100 microM ) had no effect . DB04786 ( 100 microM ) and reactive blue ( 50 microM ) decreased the number of responding cells . Removing Ca2+ from the bath diminished neither the peak in [Ca2+]i nor the percentage of responding cells , but the average [Ca2+]i increase in 1 min was significantly reduced . The results indicate that P41231 receptors are present in rat mesangial cells but it can not be excluded that there are receptors distinct from P41231 which also mediate a rise in [Ca2+]i . 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 . 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 . MicroRNAs may mediate the down-regulation of neurokinin-1 receptor in chronic bladder pain syndrome . Bladder pain syndrome ( BPS ) is a clinical syndrome of pelvic pain and urinary urgency-frequency in the absence of a specific cause . Investigating the expression levels of genes involved in the regulation of epithelial permeability , bladder contractility , and inflammation , we show that neurokinin (NK)1 and NK2 tachykinin receptors were significantly down-regulated in BPS patients . Tight junction proteins zona occludens-1 , junctional adherins molecule -1 , and occludin were similarly down-regulated , implicating increased urothelial permeability , whereas bradykinin B(1) receptor , cannabinoid receptor P21554 and muscarinic receptors M3-M5 were up-regulated . Using cell-based models , we show that prolonged exposure of P25103 to DB05875 caused a decrease of P25103 mRNA levels and a concomitant increase of regulatory micro(mi)RNAs miR-449b and miR-500 . In the biopsies of BPS patients , the same miRNAs were significantly increased , suggesting that BPS promotes an attenuation of P25103 synthesis via activation of specific miRNAs . We confirm this hypothesis by identifying 31 differentially expressed miRNAs in BPS patients and demonstrate a direct correlation between miR-449b , miR-500 , miR-328 , and miR-320 and a down-regulation of P25103 mRNA and/or protein levels . Our findings further the knowledge of the molecular mechanisms of BPS , and have relevance for other clinical conditions involving the NK1 receptor . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . DB00050 suppression test to assess the source of androgen overproduction in postmenopausal hirsutism . A 75-year-old woman presenting with recent onset hirsutism and severely elevated serum androgen levels was evaluated to assess the source of excessive androgen production . Commonly recommended hormonal stimulation and suppression tests , and the usually employed imaging techniques were non-diagnostic . In this report , we describe a new suppression test based on the use of the P30968 antagonist , cetrorelix , to determine whether androgen production was LH-dependent . DB00050 , administered in a daily dose of 250 microg subcutaneously , suppressed serum LH within 24 h and reduced serum androgen levels to normal within 48-72 h , indicating that androgen overproduction was of ovarian origin . This diagnosis was confirmed by laparoscopic ovariectomy . CONCLUSION : The cetrorelix suppression test is a simple procedure that provides valuable information regarding the source of androgen excess in postmenopausal hirsutism . Further evidence for a functional role of the glutamate receptor gene Q14832 in schizophrenia . In recent years , evidence has been accumulating indicating a major role of glutamate in the pathogenesis and pathophysiology of schizophrenia . Of particular importance in this regard are the metabotropic glutamate receptors ( GRM ) . Thus , a recently published trial of the amino acid analogue DB05096 , which exerts its effects through the activation of the glutamate receptors Q14832 / Q14416 , showed an improvement of positive and negative symptoms comparable to treatment with olanzapine . A functional variant of Q14832 has been described which modulates synaptic glutamate levels . We assessed whether this functional variant rs6465084 is related to schizophrenia in a large sample of patients and controls . We found an increased frequency of the A allele ( p=0.027 ) and the AA genotype ( p=0.024 ) in schizophrenia patients . Moreover , in an assessment of schizophrenia endophenotypes , patients of the AA genotype performed poorly in the digit symbol test , a measure of attention ( p=0.008 ) . Our results provide further evidence for the potential importance of the glutamate receptor Q14832 in schizophrenia , and indicate that the novel antipsychotic DB05096 may actually be targeting a pathogenic pathway of schizophrenia . Endocytosis mechanism of P41231 nucleotide receptor tagged with green fluorescent protein : clathrin and actin cytoskeleton dependence . Extracellular nucleotides exert a large number of physiological effects through activation of P2Y receptors . We expressed rat P41231 ( rP2Y2 ) receptor , tagged with green fluorescent protein ( GFP ) in P29320 -293 cells and visualized receptor translocation in live cells by confocal microscopy . Functional receptor expression was confirmed by determining [Ca2+]i responses . Agonist stimulation caused a time-dependent translocation of the receptor from the plasma membrane to the cytoplasm . Rearrangement of the actin cytoskeleton was observed during agonist-mediated rP2Y2-GFP receptor internalization . Colocalization of the internalized receptor with early endosomes , clathrin and lysosomes was detected by confocal microscopy . The inhibition of receptor endocytosis by either high-density medium or chlorpromazine in the presence of UTP indicates that the receptor was internalized by the clathrin-mediated pathway . The caveolin-mediated pathway was not involved . Targeting of the receptor from endosomes to lysosomes seems to involve the proteasome pathway , because proteasomal inhibition increased receptor recycling back to the plasma membrane . p38 Q96HU1 kinase regulates stem cell apoptosis in human hematopoietic failure . Myelodysplastic syndromes ( P43034 ) are clonal stem cell disorders that lead to ineffective hematopoiesis and are common causes of low blood counts in the elderly . The exact molecular mechanisms regulating increased stem apoptosis in these disorders are not well defined . p38 MAPK activation is important in regulating the growth inhibitory signals of P01375 , TGF-beta and Interferons on human hematopoiesis . Our findings show that p38 MAPK is overactivated in myelodysplasia bone marrows and regulates hematopoietic stem cell apoptosis . Inhibition of p38 MAPK by genetic or pharmacologic means decreases apoptosis and stimulates in vitro hematopoiesis from primary P43034 hematopoietic progenitors . These studies point to the potential efficacy of selective p38alpha inhibitor , DB05412 , in human bone marrow failure . Pharmacological treatment of alcohol dependence : target symptoms and target mechanisms . Alcoholism is a major public health problem and resembles , in many ways , other chronic relapsing medical conditions . At least 2 separate dimensions of its symptomatology offer targetable pathophysiological mechanisms . Systems that mediate positive reinforcement by alcohol are likely important targets in early stages of the disease , particularly in genetically susceptible individuals . In contrast , long term neuroadaptive changes caused by chronic alcohol use primarily appear to affect systems mediating negative affective states , and gain importance following a prolonged history of dependence . Feasibility of pharmacological treatment in alcoholism has been demonstrated by a first wave of drugs which consists of 3 currently approved medications , the aldehyde dehydrogenase blocker disulfiram , the opioid antagonist naltrexone ( NTX ) and the functional glutamate antagonist acamprosate ( ACM ) . The treatment toolkit is likely to be expanded in the near future . This will improve overall efficacy and allow individualized treatment , ultimately taking in account the patient 's genetic makeup . In a second wave , early human efficacy data are available for the 5HT3 antagonist ondansetron , the GABA-B agonist baclofen and the anticonvulsant topiramate . The third wave is comprised of compounds predicted to be effective based on a battery of animal models . Using such models , a short list of additional targets has accumulated sufficient preclinical validation to merit clinical development . These include the cannabinoid P21554 receptor , receptors modulating glutamatergic transmission ( Q14416 , 3 and 5 ) , and receptors for stress-related neuropeptides corticotropin releasing factor ( CRF ) , neuropeptide Y ( P01303 ) and nociceptin . Once novel treatments are developed , the field faces a major challenge to assure their delivery to patients . Improved insulin sensitivity by calorie restriction is associated with reduction of P29323 and p70S6K activities in the liver of obese Zucker rats . Calorie restriction ( CR ) improves obesity-related insulin resistance through undefined molecular mechanisms . P06213 substrate ( P41252 ) -1 serine/threonine kinases have been proposed to modulate insulin sensitivity through phosphorylation of P41252 proteins . The aim of this study is to test the hypothesis that changes in the activity of P35568 serine/threonine kinases may underlie the molecular mechanism of CR in improving insulin sensitivity . Obese and lean Zucker rats were subjected to 40 % CR or allowed to feed ad libitum ( AL ) for 20 weeks ; body weight and insulin sensitivity were monitored throughout this period . The activity of P35568 serine/threonine kinases - including JNK , P29323 , P42345 / P08133 (S6K) ( P23443 as listed in the MGI Database ) , glycogen synthase kinase 3beta ( P49841 ) , AMPK ( Q13131 as listed in the MGI Database ) , and protein kinase C ( Q04759 ) in liver tissue extracts was measured by an in vitro kinase assay using various glutathione-S-transferase ( Q86UG4 ) - P35568 fragments as substrates , while phosphorylation of P35568 and serine kinases was determined by western blotting using phosphospecific antibodies . CR in obese rats significantly reduced body weight and increased insulin sensitivity compared to AL controls . DB00133 kinase activity toward P35568 (S612) ( corresponding to S616 in human P35568 ) and P35568 (S632/635) ( corresponding to S636/639 in human P35568 ) was increased in obese rats compared to lean littermates , and was markedly decreased following CR . Concomitantly , obesity increased and CR decreased the activity of hepatic P29323 and P08133 (S6K) against P35568 . The close association between the activity of hepatic P29323 and P08133 (S6K) with insulin resistance suggests an important role for P29323 and P08133 (S6K) in the development of insulin resistance , presumably via phosphorylation of P41252 proteins . The bovine 5' AMPK gene family : mapping and single nucleotide polymorphism detection . The DB00131 -activated protein kinase ( AMPK ) family is an ancient stress response system whose primary function is regulation of cellular DB00171 . Activation of AMPK , which is instigated by environmental and nutritional stresses , initiates energy-conserving measures that protect the cell by inhibition and phosphorylation of key enzymes in energy-consuming biochemical pathways . The seven genes that comprise the bovine AMPK family were mapped in cattle by using a radiation hybrid panel . The seven genes mapped to six different cattle chromosomes , each with a LOD score greater than 10.0 . Q13131 mapped to BTA 20 , P54646 and O43741 to BTA 3 , Q9Y478 to BTA 17 , P54619 to BTA 5 , Q9UGJ0 to BTA 4 , and Q9UGI9 to BTA 2 . Five of the seven genes mapped to regions expected from human/cattle comparative maps . O43741 and Q9UGI9 , however , have not been mapped in humans . We predict these genes to be located on HSA 1 and 2 , respectively . Additionally , one synonymous and one non-synonymous single nucleotide polymorphism ( SNP ) were detected in Q9UGI9 in Bos taurus cattle . In an effort to determine ancestral origins , various herds of mixed breed cattle as well as other ruminant species were characterized for sequence variation in this region of Q9UGI9 . Owing to the physiological importance of this gene family , we believe that its individual genes are candidate genes for conferring resistance to diseases in cattle . 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 . 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 . Anandamide regulates the expression of GnRH1 , GnRH2 , and DB00644 -Rs in frog testis . DB00644 ( either GnRH1 or GnRH2 ) exerts a local activity in vertebrate testis , including human testis . Relationships between endocannabinoid ( eCB ) and DB00644 systems in gonads have never been elucidated in any species so far . To reveal a cross-talk between eCBs and DB00644 at testicular level , we characterized the expression of DB00644 ( GnRH1 and GnRH2 ) as well as P30968 ( DB00644 - Q96GN5 , -R2 , and -R3 ) mRNA in the testis of the anuran amphibian Rana esculenta during the annual sexual cycle ; furthermore , the corresponding transcripts were localized inside the testis by in situ hybridization . The possible endogenous production of the eCB , anandamide ( AEA ) , was investigated in testis by analyzing the expression of its biosynthetic enzyme , Nape-pld . Incubations of testis pieces with AEA were carried out in the postreproductive period ( June ) and in February , when a new spermatogenetic wave takes place . In June , AEA treatment significantly decreased GnRH1 and DB00644 -R2 mRNA , stimulated the transcription of GnRH2 and DB00644 - Q96GN5 , and did not affect DB00644 -R3 expression . In February , AEA treatment upregulated GnRH2 and DB00644 -R3 mRNA , downregulated DB00644 -R2 , and did not affect GnRH1 and DB00644 - Q96GN5 expression . These effects were mediated by type 1 cannabinoid receptor ( P21554 ) since they were fully counteracted by SR141716A ( DB06155 ) , a selective P21554 antagonist . In conclusion , eCB system modulates DB00644 activity in frog testis during the annual sexual cycle in a stage-dependent fashion . 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 . Peripheral endocannabinoid system-mediated actions of rimonabant on growth hormone secretion are ghrelin-dependent . The somatotroph axis is a crucial pathway regulating metabolism . Despite the fact that the endocannabinoid system has been also revealed as a potent modulator of energy homeostasis , little information is available concerning a putative interaction between these two systems . The aim of the present study was to determine the in vivo effects of the blockade of the cannabinoid receptor type 1 ( P21554 ) over growth hormone ( GH ) secretion using the P21554 antagonist rimonabant . The results obtained show that the blockade of the P21554 peripheral receptor by i.p. injection of rimonabant significantly inhibited pulsatile GH secretion . Similarly , it was found that this injection significantly decreased ghrelin-induced GH secretion without any effect on growth hormone-releasing hormone ( P01286 ) -induced GH discharge . In situ hybridisation showed that the peripheral blockade of P21554 did not affect hypothalamic somatostatin mRNA levels ; however , P01286 mRNA expression was significantly decreased . The blockade of the vagus nerve signal by surgical vagotomy eliminated the inhibitory action of rimonabant on P01286 mRNA and consequently on GH . On the other hand , the central P21554 blockade by i.c.v. rimonabant treatment was unable to reproduce the effect of peripheral blockade on P01286 mRNA , nor the GH response to ghrelin . In conclusion , the data reported in the present study establish , from a physiological point of view , the existence of a novel mechanism of GH regulation implicating the action of the cannabinoid receptor on the somatotroph axis . The effects of a cyclooxygenase-2 ( P35354 ) expression and inhibition on human uveal melanoma cell proliferation and macrophage nitric oxide production . BACKGROUND : P35354 ( P35354 ) expression has previously been identified in uveal melanoma although the biological role of P35354 in this intraocular malignancy has not been elucidated . This study aimed to investigate the effect of a P35354 inhibitor on the proliferation rate of human uveal melanoma cells , as well as its effect on the cytotoxic response of macrophages . METHODS : Human uveal melanoma cell lines were transfected to constitutively express P35354 and the proliferative rate of these cells using two different methods , with and without the addition of Amfenac , was measured . DB00435 production by macrophages was measured after exposure to melanoma-conditioned medium from both groups of cells as well as with and without Amfenac , the active metabolite of DB06802 . RESULTS : Cells transfected to express P35354 had a higher proliferation rate than those that did not . The addition of Amfenac significantly decreased the proliferation rate of all cell lines . DB00435 production by macrophages was inhibited by the addition of melanoma conditioned medium , the addition of Amfenac partially overcame this inhibition . CONCLUSION : Amfenac affected both P35354 transfected and non-transfected uveal melanoma cells in terms of their proliferation rates as well as their suppressive effects on macrophage cytotoxic activity .

MH_train_1165

MH_train_1165

MH_train_1165

interacts_with DB00850?

multiple_choice

Immunophenotypic profile and role of adhesion molecules in splenic marginal zone lymphoma with bone marrow involvement . Splenic Marginal Zone Lymphoma ( SMZL ) , with or without villous lymphocytes ( VL+/- ) , is a low-grade lymphoproliferative disorder with constant involvement of the bone marrow ( BM ) . Different BM infiltration patterns , mainly intra-sinusoidal , interstitial and nodular , have been described . Adhesion molecules ( AMs ) constitute a heterogeneous group of antigenic receptors playing a major role in leukocyte recruitment , in lymphocyte homing and in cellular-mediated immune response . Evolution and pattern of the BM infiltrate could be influenced by a variable expression of AM on SMZL lymphocytes . The degree and pattern of BM infiltration and the immunohistochemical expression of AM ( H- P62158 , P20273 , P14151 , Q14242 , P16581 , P05362 , P19320 and Beta-1 integrin ) among the different infiltration patterns were evaluated in BM biopsies of 38 patients with SMZL and graded according to a semi-quantitative score ranging from 0-4 and based on the percentage of positive cells . An intra-sinusoidal infiltration was constantly observed , alone or in conjunction with other patterns . H- P62158 and P20273 showed a moderate-to-high degree of positivity in the intra-sinusoidal infiltrate ( median expression grade-3 ) and were expressed in the neoplastic lymphocytes independently from the pattern . Q14242 was mostly expressed in the perisinusoidal region and in case of interstitial infiltration ( grade-2 ) . P05362 and P19320 were selectively expressed in the nodules as a reticular meshwork located in the core region ( grade-2 ) ; P19320 was also expressed in the perinodular endothelia . P16581 , P14151 and beta-1 integrin proved constantly negative . These data suggest that different expression of AM can influence the modality of BM infiltration in SMZL . Characterization of DB05005 -B , an orally bioavailable antagonist of the P51686 chemokine receptor , for treatment of inflammatory bowel disease . The chemokine system represents a diverse group of G protein-coupled receptors responsible for orchestrating cell recruitment under both homeostatic and inflammatory conditions . Chemokine receptor 9 ( P51686 ) is a chemokine receptor known to be central for migration of immune cells into the intestine . Its only ligand , O15444 , is expressed at the mucosal surface of the intestine and is known to be elevated in intestinal inflammation . To date , there are no reports of small-molecule antagonists targeting P51686 . We report , for the first time , the discovery of a small molecule , DB05005 -B , which is an orally bioavailable , selective , and potent antagonist of human P51686 . DB05005 -B inhibited P51686 -mediated Ca(2+) mobilization and chemotaxis on Molt-4 cells with IC(50) values of 5.4 and 3.4 nM , respectively . In the presence of 100 % human serum , DB05005 -B inhibited P51686 -mediated chemotaxis with an IC(50) of 33 nM , and the addition of α1-acid glycoprotein did not affect its potency . DB05005 -B inhibited chemotaxis of primary P51686 -expressing cells to O15444 with an IC(50) of 6.8 nM . DB05005 -B was an equipotent inhibitor of O15444 -directed chemotaxis of both splice forms of P51686 ( CCR9A and CCR9B ) with IC(50) values of 2.8 and 2.6 nM , respectively . DB05005 -B also inhibited mouse and rat P51686 -mediated chemotaxis . Inhibition of P51686 with DB05005 -B results in normalization of Crohn 's disease such as histopathology associated with the P01375 (ΔARE) mice . Analysis of the plasma level of drug associated with this improvement provides an understanding of the pharmacokinetic/pharmacodynamic relationship for P51686 antagonists in the treatment of intestinal inflammation . 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 . Phase 1 clinical results with DB05465 ( MLN518 ) , a novel P36888 antagonist , in patients with acute myelogenous leukemia or high-risk myelodysplastic syndrome : safety , pharmacokinetics , and pharmacodynamics . Tandutinib ( MLN518/CT53518 ) is a novel quinazoline-based inhibitor of the type III receptor tyrosine kinases : P07333 -like tyrosine kinase 3 ( P36888 ) , platelet-derived growth factor receptor ( P09619 ) , and P10721 . Because of the correlation between P36888 internal tandem duplication ( ITD ) mutations and poor prognosis in acute myelogenous leukemia ( AML ) , we conducted a phase 1 trial of DB05465 in 40 patients with either AML or high-risk myelodysplastic syndrome ( P43034 ) . Tandutinib was given orally in doses ranging from 50 mg to 700 mg twice daily The principal dose-limiting toxicity ( DLT ) of DB05465 was reversible generalized muscular weakness , fatigue , or both , occurring at doses of 525 mg and 700 mg twice daily . Tandutinib 's pharmacokinetics were characterized by slow elimination , with achievement of steady-state plasma concentrations requiring greater than 1 week of dosing . Western blotting showed that DB05465 inhibited phosphorylation of P36888 in circulating leukemic blasts . Eight patients had P36888 -ITD mutations ; 5 of these were evaluable for assessment of DB05465 's antileukemic effect . Two of the 5 patients , treated at 525 mg and 700 mg twice daily , showed evidence of antileukemic activity , with decreases in both peripheral and bone marrow blasts . Tandutinib at the MTD ( 525 mg twice daily ) should be evaluated more extensively in patients with AML with P36888 -ITD mutations to better define its antileukemic activity . 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 . Differential effects of epratuzumab on peripheral blood B cells of patients with systemic lupus erythematosus versus normal controls . OBJECTIVE : B lymphocytes have been implicated in the pathogenesis of lupus and other autoimmune diseases , resulting in the introduction of B cell-directed therapies . DB04958 , a humanised anti- P20273 monoclonal antibody , is currently in clinical trials , although its effects on patients ' B cells are not completely understood . METHODS : This study analysed the in vivo effect of epratuzumab on peripheral B cell subsets in 12 patients with systemic lupus erythematosus , and also addressed the in vitro effects of the drug by analysing anti-immunoglobulin-induced proliferation of isolated B cells obtained from the peripheral blood of 11 additional patients with lupus and seven normal subjects . RESULTS : Upon treatment , a pronounced reduction of P26842 (-) B cells and P20273 surface expression on P26842 (-) B cells was observed , suggesting that these cells , which mainly comprise naïve and transitional B cells , are preferentially targeted by epratuzumab in vivo . The results of in vitro studies indicate additional regulatory effects of the drug by reducing the enhanced activation and proliferation of anti-immunoglobulin-stimulated lupus B cells after co-incubation with P29965 or CpG . DB04958 inhibited the proliferation of B cells from patients with systemic lupus erythematosus but not normal B cells under all culture conditions . CONCLUSIONS : DB04958 preferentially modulates the exaggerated activation and proliferation of B cells from patients with lupus in contrast to normal subjects , thus suggesting that epratuzumab might offer a new therapeutic option for patients with systemic lupus erythematosus , as enhanced B cell activation is a hallmark of this disease . Dual acylation and lipid raft association of Src-family protein tyrosine kinases are required for P48061 / P48061 -mediated chemotaxis in the Jurkat human T cell lymphoma cell line . Chemokines play pivotal roles in regulating a wide variety of biological processes by modulating cell migration and recruitment . Deregulation of chemokine signaling can alter cell recruitment , contributing to the pathogenic states associated with autoimmune disease , inflammatory disorders , and sepsis . During chemotaxis , lipid rafts and their resident signaling molecules have been demonstrated to partition to different parts of the cell . Herein , we investigated the role of lipid raft resident Src-family kinases ( SFK ) in stromal cell-derived factor 1/ P48061 -mediated chemotaxis . We have shown that Lck-deficient J. P62158 1.6 cells are defective in P48061 -mediated chemotaxis in contrast to their parental counterpart , Jurkat cells . Ectopic expression of the SFK hematopoietic cell kinase ( Hck ) in J. P62158 1.6 cells reconstituted P48061 responsiveness . The requirement of lipid raft association of SFK was assessed using both isoforms of Hck : the dually acylated p59(Hck) isoform that is targeted to lipid rafts and the monoacylated p61(Hck) isoform that is nonraft-associated . We have shown using several gain and loss of acylation alleles that dual acylation of Hck was required for P48061 -mediated chemotaxis in J. P62158 1.6 cells . These results highlight the importance of the unique microenvironment provided by lipid rafts and their specific contribution in providing specificity to P48061 signaling . Separation and purification of the tonoplast ATPase and pyrophosphatase from plants with constitutive and inducible Crassulacean acid metabolism . Tonoplast vesicles were isolated from Kalanchoe daigremontiana Hamet et Pierrer de la Bâthie and Mesembryanthemum crystallinum L. , exhibiting constitutive and inducible crassulacean acid metabolism ( P62158 ) , respectively . Membrane-bound proteins were detergent-solubilized with 2 % of Triton X-100 . During P62158 induction in M. crystallinum , ATPase activity increases four-fold , whereas pyrophosphatase activity decreases somewhat . With all plants , ATPase and pyrophosphatase could be separated by size-exclusion chromatography ( SEC , Sephacryl S 400 ) , and the ATPase was further purified by diethylaminoethyl-ion-exchange chromatography . Sodium-dodecyl-sulfate electrophoresis of the SEC fractions from K. daigremontiana containing maximum ATPase activity separates several protein bands , indicating subunits of 72 , 56 , 48 , 42 , 28 , and 16 kDa . Purified ATPase from M. crystallinum in the P01024 and P62158 states shows a somewhat different protein pattern . With M. crystallinum , an increase in DB00171 -hydrolysis and changes in the subunit composition of the native enzyme indicate that the change from the P01024 to the P62158 state is accompanied by de-novo synthesis and by structural changes of the tonoplast ATPase . 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 . 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 . Effect of type-selective inhibitors on cyclic nucleotide phosphodiesterase activity and insulin secretion in the clonal insulin secreting cell line BRIN-BD11 . 1. The cyclic nucleotide phosphodiesterases ( PDEs ) present in an insulin secreting cell line , BRIN - BD11 , were characterized using calcium/calmodulin , DB01277 , isoenzyme-selective PDE inhibitors and RT - PCR . 2 . P62158 activated cyclic AMP or cyclic GMP PDE activity in pellet and was 3 fold ( P=0.002 ) more potent in activating cyclic nucleotide hydrolysis in pellet compared with supernatant fractions . 3 . The PDE1/ O76074 inhibitor zaprinast inhibited both cyclic AMP and cyclic GMP PDE activity in both pellet and supernatant fractions of cell homogenates by a maximum of around 25 % ( IC(50) 1 - 5 microM ) , while rolipram ( DB05876 selective ) inhibited only cyclic AMP hydrolysis . 4 . The PDE3-selective inhibitors Org 9935 ( 0.02 - 10 microM ) and siguazodan ( 0.1 - 10 microM ) inhibited cyclic AMP PDE activity in the pellet but not the supernatant fractions of cell homogenates , with a maximum inhibition of about 30 % . DB01277 ( 2 - 7.5 ng ml(-1) ) potently augmented this PDE activity . 5 . RT - PCR using specific primers for Q13370 , but not for Q14432 , amplified , from BRIN - BD11 cell total RNA , a 351 base pair product that was > 97 % homologous with rat adipose tissue Q13370 . 6 . DB07954 , Org 9935 , siguazodan and rolipram ( 1 - 50 microM ) , but not zaprinast , each augmented glucose-induced insulin secretion in the presence of 16.7 mM but not 1 mM glucose . 7 . These findings , in a clonal insulin secreting cell line , are consistent with an important role for Q13370 in regulating the pool of cyclic AMP relevant to the modulation of glucose-induced insulin secretion . Inhibition of PDE3 , DB05876 and PDE7 potentiates glucocorticoid-induced apoptosis and overcomes glucocorticoid resistance in CEM T leukemic cells . Stimulation of the DB02527 signaling pathway has been shown to induce apoptosis and augment the effects of glucocorticoids in inducing apoptosis in leukemic cells . We recently reported that in primary B cell chronic lymphocytic leukemic ( B-CLL ) cells , apoptosis could be induced by stimulating the DB02527 signaling pathway with a phosphodiesterase4 ( DB05876 ) inhibitor alone ; while in contrast , in the CEM T leukemic cell line , DB05876 inhibitors alone were ineffective , and concurrent stimulation of adenylyl cyclase was required to see effects [ Tiwari et al. ( 2005 ) ] . We report here that in the CEM and Jurkat T leukemic cell lines , the most abundantly expressed PDEs are Q13370 , P27815 , Q08499 , Q13946 , and O60658 . Selective inhibition of PDE3 , DB05876 or PDE7 alone produces little effect on cell viability , but inhibition of all three of these PDEs together dramatically enhances glucocorticoid-induced apoptosis in CEM cells , and overcomes glucocorticoid resistance in a glucocorticoid-resistant CEM cell line . These studies indicate that for some leukemic cell types , a desired therapeutic effect may be achieved by inhibiting more than one form of PDE . Inhibition of human steroid 5beta-reductase ( P51857 ) by finasteride and structure of the enzyme-inhibitor complex . The Delta(4)-3-ketosteroid functionality is present in nearly all steroid hormones apart from estrogens . The first step in functionalization of the A-ring is mediated in humans by steroid 5alpha- or 5beta-reductase . DB01216 is a mechanism-based inactivator of 5alpha-reductase type 2 with subnanomolar affinity and is widely used as a therapeutic for the treatment of benign prostatic hyperplasia . It is also used for androgen deprivation in hormone-dependent prostate carcinoma , and it has been examined as a chemopreventive agent in prostate cancer . The effect of finasteride on steroid 5beta-reductase ( P51857 ) has not been previously reported . We show that finasteride competitively inhibits P51857 with low micromolar affinity but does not act as a mechanism-based inactivator . The structure of the P51857 .NADP(+)*finasteride complex determined at 1.7 A resolution shows that it is not possible for NADPH to reduce the Delta(1-2)-ene of finasteride because the cofactor and steroid are not proximal to each other . The P01024 -ketone of finasteride accepts hydrogen bonds from the catalytic residues DB00135 -58 and DB00142 -120 in the active site of P51857 , providing an explanation for the competitive inhibition observed . This is the first reported structure of finasteride bound to an enzyme involved in steroid hormone metabolism . SDF-1alpha/ P48061 enhances GABA and glutamate synaptic activity at serotonin neurons in the rat dorsal raphe nucleus . The serotonin ( 5-hydroxytryptamine ; 5-HT ) system has a well-characterized role in depression . Recent reports describe comorbidities of mood-immune disorders , suggesting an immunological component may contribute to the pathogenesis of depression as well . Chemokines , immune proteins which mediate leukocyte trafficking , and their receptors are widely distributed in the brain , mediate neuronal patterning , and modulate various neuropathologies . The purpose of this study was to investigate the neuroanatomical relationship and functional impact of the chemokine stromal cell-derived factor-1alpha/ P48061 and its receptor , P61073 , on the serotonin dorsal raphe nucleus ( DRN ) system in the rat using anatomical and electrophysiological techniques . Immunohistochemical analysis indicates that over 70 % of 5-HT neurons colocalize with P48061 and P61073 . At a subcellular level , P48061 localizes throughout the cytoplasm whereas P61073 concentrates to the outer membrane and processes of 5-HT neurons . P48061 and P61073 also colocalize on individual DRN cells . Furthermore , electrophysiological studies demonstrate P48061 depolarization of 5-HT neurons indirectly via glutamate synaptic inputs . P48061 also enhances the frequency of spontaneous inhibitory and excitatory postsynaptic currents ( sIPSC and sEPSC ) . P48061 concentration-dependently increases evoked IPSC amplitude and decreases evoked IPSC paired-pulse ratio selectively in 5-HT neurons , effects blocked by the P61073 antagonist DB06809 . These data indicate presynaptic enhancement of GABA and glutamate release at 5-HT DRN neurons by P48061 . Immunohistochemical analysis further shows P61073 localization to DRN GABA neurons , providing an anatomical basis for P48061 effects on GABA release . Thus , P48061 indirectly modulates 5-HT neurotransmission via GABA and glutamate synaptic afferents . Future therapies targeting P48061 and other chemokines may treat serotonin related mood disorders , particularly depression experienced by immune-compromised individuals . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . Expression of adhesion molecules and c-kit on P28906 + hematopoietic progenitor cells : comparison of cytokine-mobilized blood stem cells with normal bone marrow and peripheral blood . We assessed the expression of the adhesion molecules leukocyte function antigen-1 ( LFA-1 , CD11a ) , intercellular adhesion molecule-1 ( P05362 , CD54 ) , homing-associated cell adhesion molecule ( H- P62158 , P16070 ) , and c-kit ( stem cell factor receptor ) on the P28906 + progenitor population from the leukapheresis products of 23 patients ( LP P28906 + ) . For blood stem cell collection granulocyte colony-stimulating factor ( DB00099 ) or interleukin-3/granulocyte-macrophage colony-stimulating factor ( P08700 /GM- P04141 ) was administered after cytotoxic chemotherapy . Furthermore , bone marrow- and blood-derived P28906 + progenitor cells from 6 normal volunteers ( BM and PB P28906 + ) were analyzed . LFA-1 expression was higher on PB P28906 + ( 88.2 +/- 2.5 % , mean +/- SEM ) than on BM P28906 + ( 75.3 +/- 4.3 % ) . Following cytokine administration , LFA-1 was expressed on only 59.7 +/- 3.7 % of LP P28906 + at a low fluorescence intensity , suggesting that down-regulation of LFA-1 may facilitate the egress of cells from the bone marrow and prolong their circulation . In contrast , P05362 was weakly positive on P28906 + cells from all sources . P16070 was expressed on the vast majority of P28906 + cells ( > 95 % ) in all samples studied . The highest proportion of P28906 + cells costaining for c-kit was found in normal bone marrow ( 32.2 +/- 3.3 % ) . In normal peripheral blood and after cytokine mobilization , fewer of the P28906 + cells weakly expressed c-kit ( < 15 % ) . The low percentage and level of c-kit expression may indicate that the majority of cytokine-mobilized P28906 + cells are lineage-committed progenitor cells , as reflected by the coexpression pattern for P28907 , HLA-DR , and P20138 . 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 . Improvement in Long-Term Memory following Chronic Administration of Eryngium planum Root Extract in DB00747 Model : Behavioral and Molecular Study . Eryngium planum L . ( EP ) is as a rare medicinal plant with a lot of potentials as pharmaceutical crops . The aim of our study was to assess the effect of subchronic ( 28-fold ) administration of a 70 % ethanol extract of EP roots ( 200 mg/kg , p.o. ) on behavioral and cognitive responses in Wistar rats linked with acetylcholinesterase ( P22303 ) , butyrylcholinesterase ( BuChE ) , and beta-secretase ( P56817 -1 ) mRNA levels and P22303 and BuChE activities in the hippocampus and frontal cortex . On the last day of experiment , 30 min after the last dose of EP or DB04864 ( HU ) , scopolamine ( SC ) was given at a dose of 0.5 mg/kg b.w. intraperitoneally . The results of a passive avoidance test showed an improvement in long-term memory produced by the EP extract in both scopolamine-induced rats and control group . EP caused an insignificant inhibition of P22303 and BuChE activities in the frontal cortex and the hippocampus . EP decreased mRNA P22303 , BuChE , and P56817 -1 levels , especially in the cortex . Our results suggest that the EP extract led to the improvement of the long-term memory in rats coupled with total saponin content . The mechanism of EP action is probably complicated , since HPLC-MS analysis showed 64 chemical compounds ( phenolics , saponins ) in the extract of EP roots . 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 . Stimulation of type 1 and type 8 Ca2+/calmodulin-sensitive adenylyl cyclases by the Gs-coupled 5-hydroxytryptamine subtype 5-HT7A receptor . The neurotransmitter serotonin ( 5-hydroxytryptamine , 5-HT ) plays an important regulatory role in developing and adult nervous systems . With the exception of the 5- Q9H205 receptor , all of the cloned serotonin receptors belong to the G protein-coupled receptor superfamily . Subtypes P50406 and P34969 couple to stimulation of adenylyl cyclases through Gs and display high affinities for antipsychotic and antidepressant drugs . In the brain , mRNA for P50406 is found at high levels in the hippocampus , striatum , and nucleus accumbens . P34969 mRNA is most abundant in the hippocampus , neocortex , and hypothalamus . To better understand how serotonin might control DB02527 levels in the brain , we coexpressed P50406 or 5-HT7A receptors with specific isoforms of adenylyl cyclase in P29320 293 cells . The P50406 receptor functioned as a typical Gs-coupled receptor in that it stimulated O95622 , a Gs-sensitive adenylyl cyclase , but not Q99440 or P40145 , calmodulin ( P62158 ) -stimulated adenylyl cyclases that are not activated by Gs-coupled receptors in vivo . Surprisingly , serotonin activation of 5-HT7A stimulated Q99440 and P40145 by increasing intracellular Ca2+ . 5-HT also increased intracellular Ca2+ in primary neuron cultures . These data define a novel mechanism for the regulation of intracellular DB02527 by serotonin .

MH_train_1166

MH_train_1166

MH_train_1166

interacts_with DB00333?

multiple_choice

Chromosome mapping of the human arrestin ( SAG ) , beta-arrestin 2 ( P32121 ) , and beta-adrenergic receptor kinase 2 ( P35626 ) genes . Two types of proteins play a major role in determining homologous desensitization of G-coupled receptors : beta-adrenergic receptor kinase ( beta ARK ) , which phosphorylates the agonist-occupied receptor and its functional cofactor , beta-arrestin . Both beta ARK and beta-arrestin are members of multigene families . The family of G-protein-coupled receptor kinases includes rhodopsin kinase , beta O14965 , beta Q96GD4 , IT11-A ( P32298 ) , P34947 , and P43250 . The arrestin/beta-arrestin gene family includes arrestin ( also known as S-antigen ) , beta-arrestin 1 , and beta-arrestin 2 . Here we report the chromosome mapping of the human genes for arrestin ( SAG ) , beta-arrestin 2 ( P32121 ) , and beta Q96GD4 ( P35626 ) by fluorescence in situ hybridization ( Q5TCZ1 ) . Q5TCZ1 results confirmed the assignment of the gene coding for arrestin ( SAG ) to chromosome 2 and allowed us to refine its localization to band q37 . The gene coding for beta-arrestin 2 ( P32121 ) was mapped to chromosome 17p13 and that coding for beta Q96GD4 ( P35626 ) to chromosome 22q11 . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Pediatric phase I trial and pharmacokinetic study of DB05220 , an investigational oral selective small-molecule inhibitor of O14965 : a Children 's Oncology Group Phase I Consortium study . PURPOSE : DB05220 , a selective small-molecule inhibitor of O14965 , has activity in a broad range of preclinical pediatric cancer models . We conducted a phase I trial in children with refractory/recurrent solid tumors to define the maximum-tolerated dose , toxicities , and pharmacokinetic properties of DB05220 . EXPERIMENTAL DESIGN : DB05220 was administered orally either once daily or divided twice daily for seven days , every 21 days . Using a rolling-six design , four dose levels ( 45 , 60 , 80 , and 100 mg/m(2)/day ) were evaluated on the once-daily schedule , and two dose levels ( 60 and 80 mg/m(2)/d ) on the twice-daily schedule . Pharmacokinetic studies were conducted with the initial dose and trough drug concentrations also measured at the steady state . RESULTS : Thirty-seven patients were enrolled . On the once-daily dosing schedule , myelosuppression was dose limiting in three of four patients at 100 mg/m(2) , and one of six patients had dose-limiting mood alteration at 80 mg/m(2) . At 45 mg/m(2) , one of six patients experienced dose-limiting mucositis . Mucositis and myelosuppression were dose limiting at 80 mg/m(2) on the twice-daily schedule , and one of five patients at 60 mg/m(2) on the twice-daily schedule experienced a dose-limiting alkaline phosphatase . Five of 11 patients experienced hand-foot-skin syndrome with twice-daily dosing versus one of 21 after once-daily dosing . There was one partial response and six with prolonged stable disease among 33 evaluable subjects . CONCLUSION : The twice-daily dose regimen is well tolerated in adults ; however , children experienced a greater frequency of myelosuppression and hand-foot-skin syndrome on this schedule . Children tolerated a higher dose and the recommended pediatric phase II dose is 80 mg/m(2)/d once daily for seven days . Inducible P35354 -dependent apoptosis in human ovarian cancer cells . DB02709 is a naturally occurring trihydroxyl-diphenylethylene compound that has been shown experimentally to have beneficial effects in the treatment of cancer and cardiovascular disease . DB02709 induces programmed cell death ( apoptosis ) in these cells and activates important signal transducing proteins including extracellular signal-regulated kinases ( ERKs ) 1 and 2 in cancer cells . DB02709 also causes nuclear accumulation of the enzyme cyclooxygenase ( P36551 ) -2 and of the oncogene suppressor protein , p53 . We have studied the molecular basis of the anticancer actions of resveratrol using human ovarian carcinoma ( OVCAR-3 ) cells . Our findings include the following : ( i ) nuclear accumulation of P35354 in resveratrol-treated cells is blocked by the P27361 /2 inhibitor , PD98059 ; ( ii ) an inhibitor of P35354 activity , NS398 , prevents accumulation of P27361 /2 , P35354 , activated p53 and small ubiquitin-like modifier ( P63165 ) in the nucleus ; ( iii ) apoptosis , quantitated by nucleosome enzyme-linked immunosorbent assay and the nuclear abundance of the pro-apoptotic protein , BcL-xs , were inhibited by NS398 . This finding implicates nuclear P35354 in p53-mediated apoptosis induced by resveratrol . Sumoylation is important to stabilization of p53 and a P35354 - P63165 interaction suggests sumoylation of P35354 in resveratrol-treated cells and ( iv ) chromatin immunoprecipitation studies showed binding of induced nuclear P35354 to the promoter region of PIG3 and Bax , pro-apoptotic gene targets of transcriptionally active p53 . Nuclear accumulation of activated P27361 /2 and sumolyated P35354 are essential to resveratrol-induced pSer-15-p53-mediated apoptosis in human ovarian cancer cells . 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 . 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 . P43490 , an adipocytokine with proinflammatory and immunomodulating properties . Adipocytokines are mainly adipocyte-derived cytokines regulating metabolism and as such are key regulators of insulin resistance . Some adipocytokines such as adiponectin and leptin affect immune and inflammatory functions . P43490 ( pre-B cell colony-enhancing factor ) has recently been identified as a new adipocytokine affecting insulin resistance by binding to the insulin receptor . In this study , we show that recombinant visfatin activates human leukocytes and induces cytokine production . In P08571 (+) monocytes , visfatin induces the production of IL-1beta , P01375 , and especially P05231 . Moreover , it increases the surface expression of costimulatory molecules CD54 , P25942 , and P33681 . P43490 -stimulated monocytes show augmented FITC-dextran uptake and an enhanced capacity to induce alloproliferative responses in human lymphocytes . P43490 -induced effects involve p38 as well as Q02750 pathways as determined by inhibition with MAPK inhibitors and we observed activation of NF-kappaB . In vivo , visfatin induces circulating P05231 in BALB/c mice . In patients with inflammatory bowel disease , plasma levels of visfatin are elevated and its mRNA expression is significantly increased in colonic tissue of Crohn 's and ulcerative colitis patients compared with healthy controls . Macrophages , dendritic cells , and colonic epithelial cells might be additional sources of visfatin as determined by confocal microscopy . P43490 can be considered a new proinflammatory adipocytokine . 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 . Mechanism of inhibition of the P42262 AMPA receptor channel opening by talampanel and its enantiomer : the stereochemistry of the 4-methyl group on the diazepine ring of 2,3-benzodiazepine derivatives . Stereoselectivity of 2,3-benzodiazepine compounds provides a unique way for the design of stereoisomers as more selective and more potent inhibitors as drug candidates for treatment of the neurological diseases involving excessive activity of AMPA receptors . Here we investigate a pair of enantiomers known as DB04982 and its ( + ) counterpart about their mechanism of inhibition and selectivity toward four AMPA receptor subunits or P42261 -4 . We show that DB04982 is the eutomer with the endismic ratio being 14 for the closed-channel and 10 for the open-channel state of P42262 . Kinetic evidence supports that DB04982 is a noncompetitive inhibitor and it binds to the same site for those 2,3-benzodiazepine compounds with the C-4 methyl group on the diazepine ring . This site , which we term as the " M " site , recognizes preferentially those 2,3-benzodiazepine compounds with the C-4 methyl group being in the R configuration , as in the chemical structure of DB04982 . Given that DB04982 inhibits P42261 and P42262 , but is virtually ineffective on the P42263 and P48058 AMPA receptor subunits , we hypothesize that the " M " site(s) on P42261 and P42262 to which DB04982 binds is different from that on P42263 and P48058 . If the molecular properties of the AMPA receptors and DB04982 are used for selecting an inhibitor as a single drug candidate for controlling the activity of all AMPA receptors in vivo , DB04982 is not ideal . Our results further suggest that addition of longer acyl groups to the N-3 position should produce more potent 2,3-benzodiazepine inhibitors for the " M " site . Altered long-term corticostriatal synaptic plasticity in transgenic mice overexpressing human CU/ZN superoxide dismutase ( GLY(93) --> ALA ) mutation . Apart from the extensive loss of motor neurons , degeneration of midbrain dopaminergic cells has been described in both familial and sporadic forms of amyotrophic lateral sclerosis ( P35858 ) . Mice overexpressing the mutant human Cu/Zn superoxide dismutase ( P00441 ) show an P35858 -like phenotype in that they show a progressive death of motor neurons accompanied by degeneration of dopaminergic cells . To describe the functional alterations specifically associated with this dopaminergic dysfunction , we have investigated the corticostriatal synaptic plasticity in mice overexpressing the human P00441 ( P00441 + ) and the mutated ( DB00145 (93) --> Ala ) form ( G93A+ ) of the same enzyme . We show that repetitive stimulation of the corticostriatal pathway generates long-term depression ( LTD ) in P00441 + mice and in control ( G93A-/ P00441 - ) animals , whereas in G93A+ mice the same stimulation generates an N-methyl-D-aspartic acid receptor-dependent long-term potentiation . No significant alterations were found in the intrinsic membrane properties of striatal medium spiny neurons and basal corticostriatal synaptic transmission of G93A+ mice . Bath perfusion of dopamine or the P14416 agonist quinpirole restored LTD in G93A+ mice . Consistent with these in vitro results , habituation of locomotor activity and striatal-dependent active avoidance learning were impaired in G93A+ mice . Thus , degeneration of dopaminergic neurons in the substantia nigra of G93A+ mice causes substantial modifications in striatal synaptic plasticity and related behaviors , and may be a cellular substrate of the extrapyramidal motor and cognitive disorders observed in familial and sporadic P35858 . The role of heat shock proteins in Amyotrophic Lateral Sclerosis : The therapeutic potential of DB05025 . DB05025 is a hydroxylamine derivative , a group of compounds which have unique properties as co-inducers of heat shock protein expression , but only under conditions of cellular stress . DB05025 has been found to be neuroprotective in a number of neurodegenerative disease models , including Amyotrophic Lateral Sclerosis ( P35858 ) , and in mutant Superoxide Dismutase 1 ( P00441 ) mice that model P35858 , DB05025 rescues motor neurons , improves neuromuscular function and extends lifespan . The therapeutic potential of DB05025 is currently under investigation in a Phase II clinical trial for P35858 patients with P00441 mutations . In this review we summarize the evidence for the neuroprotective effects of enhanced heat shock protein expression by DB05025 and other inducers of the Heat Shock Response . P35858 is a complex , multifactorial disease affecting a number of cell types and intracellular pathways . Cells and pathways affected by P35858 pathology and which may be targeted by a heat shock protein-based therapy are also discussed in this review . For example , protein aggregation is a characteristic pathological feature of neurodegenerative diseases including P35858 . Enhanced heat shock protein expression not only affects protein aggregation directly , but can also lead to more effective clearance of protein aggregates via the unfolded protein response , the proteasome-ubiquitin system or by autophagy . However , compounds such as DB05025 have effects beyond targeting protein mis-handling and can also affect additional pathological mechanisms such as oxidative stress . Therefore , by targeting multiple pathological mechanisms , compounds such as DB05025 may be particularly effective in the development of a disease-modifying therapy for P35858 and other neurodegenerative disorders . P0DMS8 is a critical mediator in LPS-induced pulmonary inflammation . DB00640 receptor A(3) ( A(3) ) regulates directed movement of polymorphonuclear cells ( PMNs ) to sites of inflammation and has been implicated as a relevant mediator in models of inflammatory diseases . Here , we sought to characterize the role of A(3) in a murine model of lung inflammation . Initial studies revealed that pulmonary A(3) transcript levels were elevated following LPS exposure in vivo . In addition , inhalation of LPS increased the accumulation of PMNs in wild-type and A(3)(-/-) mice in all lung compartments . Pretreatment with the specific A(3)-agonist Cl- DB05511 significantly decreased migration of PMNs into lung interstitium and alveolar air space of wild-type mice but not of A(3)(-/-) mice . Lower PMN counts were associated with reduced levels of P01375 -α and P05231 in the alveolar space of wild-type mice that received Cl- DB05511 . In addition , Cl- DB05511 attenuated LPS-induced microvascular permeability in wild-type mice as assessed by the extravasation of Evans blue . In pulmonary microvascular endothelial cells , Cl- DB05511 reduced LPS-induced cytoskeletal remodeling and cell retraction , consistent with a specific role of A(3) for maintaining endothelial integrity . Migratory activity of human PMNs across an endothelial or epithelial monolayer was reduced when A(3) was activated on PMNs . Studies in chimeric mice , however , revealed that Cl- DB05511 required A(3) on both hematopoietic and nonhematopoietic cells to reduce transmigration in vivo . Together , our results shed new light on the role of A(3) in LPS-induced PMN trafficking in the lung and suggest pharmacological modulation of A(3)-dependent pathways as a promising approach in lung inflammation . 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 . Regulation of P27361 /2 phosphorylation by acute and chronic morphine - implications for the role of DB02527 -responsive element binding factor ( CREB ) -dependent and Ets-like protein-1 ( Elk-1 ) -dependent transcription ; small interfering RNA-based strategy . Extracellular signal-regulated kinases ( ERKs ) have been shown to be activated by opioids and functionally linked to addiction . Morphine-associated changes in P29323 activity seem to be the characteristic features of opioid action . In this study , we observed a rapid and severe increase in P27361 /2 activity after a 5 min morphine treatment of P29320 -MOR cells ( transfected with the rat mu-opioid receptor P35372 ) expressing mu-opioid receptor . Cellular adaptations to chronic ( 72 h ) morphine treatment were manifested by a slight and sustained increase in P27361 /2 activity . Withdrawal caused by an opioid receptor antagonist - naloxone - attenuated phosphorylation of P27361 /2 . Little information is available on the precise mechanism of P29323 activity regulation . Using RNA interference technology , we generated stably transfected cells with silenced expression of DB02527 -responsive element binding factor ( CREB ) and Ets-like protein-1 ( Elk-1 ) transcription factors , which are known targets for activated P27361 /2 . In these cells , P27361 /2 activity regulation was altered . Silencing of CREB or Elk-1 significantly increased P29323 activation observed after 5 min of morphine stimulation . The initial level of activated ERKs in these cells was also augmented . Moreover , the cellular response to withdrawal signals and chronic opioid treatment was diminished . These differences suggest that both CREB-dependent and Elk-1-dependent transcription contribute to the expression of proteins regulating morphine-induced P29323 activity ( particular phosphatases , upstream kinases or their activatory proteins ) . The genes encoding the glutamate receptor subunits Q16099 and Q16478 ( Q16099 and Q16478 ) are located on separate chromosomes in human , mouse , and rat . The chromosomal localization of the human and rat genes encoding the kainate-preferring glutamate receptor subunits Q16099 and Q16478 ( Q16099 and Q16478 , respectively ) was determined by Southern analysis of rat x mouse and human x mouse somatic cell hybrid panels and by fluorescence in situ hybridization . The localization of the mouse genes ( Grik4 and Grik5 ) was established by interspecific backcross mapping . Q16099 and Q16478 are located on separate chromosomes ( Chrs ) in all species . Q16099 mapped to human Chr 11q22.3 , mouse Chr 9 , and rat Chr 8 . Q16478 mapped to human Chr 19q13.2 , mouse Chr 7 , and rat Chr 1 . The genes encoding the ( R,S ) -alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ( AMPA ) -preferring subunit P48058 , or GluRD ( P48058 ) , the neural cell adhesion molecule ( P13591 ) , the D2 dopamine receptor ( P14416 ) , and the Thy-1 cell surface antigen ( P04216 ) have all been previously mapped to the human Chr 11q22 region . The mapping of the human Q16099 and Q16478 genes confirms and extends the relationship between human Chr 11 and mouse Chr 9 and also human Chr 19 and mouse Chr 7 . Q16099 is the fifth gene shared by human Chr 11 and rat Chr 8 , whereas Q16478 is 1 out of the 12 genes that are located on both human Chr 19 and rat Chr 1 . Our data extend the conserved synteny established between certain human , mouse , and rat Chrs . 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 . DB00094 -regulated gene expression profiles in ovarian tumours and normal ovaries . Development , growth and function of the ovary are controlled by endocrine and paracrine signals . These may also influence the development of ovarian cancer . The aim of this study was to identify the key molecular markers of the unregulated growth and hormone synthesis seen in ovarian tumours , particularly in granulosa cell tumours ( GCT ) . Genes used in this study were chosen on the basis of our understanding of growth and differentiation in the normal ovary . We sought to define the patterns of gene expression in a panel of epithelial and stromal ovarian tumours . Expression was determined by RT-PCR using gene-specific primers for the DB00094 receptor ( P23945 ) ; the DB00094 early response genes : regulatory subunit of protein kinase A ( RII-beta ) , cyclin D2 ( cycD2 ) and sgk ; and late response markers : cyclooxygenase-2 ( P35354 ) and the LH receptor ( LHR ) . The GCT had high expression of P23945 compared with normal ovaries and the other tumours . cycD2 and RII-beta and P35354 genes were also highly expressed in the GCT. sgk and LHR expression was lower in all of the tumours than in normal ovaries . Serous cystadenocarcinomas also had an unexpectedly high expression of P35354 . Comparison of the gene expression profiles between each tumour group suggests a molecular phenotype for GCT that is similar to that reported for DB00094 stimulated pre-ovulatory granulosa cells . 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 . DB04901 ( IDEC ) . IDEC is developing a PRIMATIZED-anti- P33681 antibody ( DB04901 ) for the treatment of autoimmune and inflammatory diseases , such as psoriasis and rheumatoid arthritis . It is currently undergoing phase II trials in patients with psoriasis [ 395813 ] . A randomized , blind , placebo-controlled , multiple-dose phase II study was initiated in January 2001 to evaluate the potential clinical activity and safety of DB04901 in patients with moderate-to-severe psoriasis [ 395813 ] . The antibody targets the P33681 antigen on the surface of antigen-presenting cells that normally interact with T-cells to initiate an immune response . Antibodies directed at P33681 may be useful in preventing unwanted immune responses in autoimmune diseases such as systemic lupus erythematosus , idiopathic thrombocytopenic purpura as well as transplant rejection [ 178382 ] , [ 178929 ] . PRIMATIZED antibodies , genetically engineered from cynomolgus macaque monkey and human components , are structurally indistinguishable from human antibodies . They may , therefore , be less likely to cause adverse reactions in humans , making them potentially suited for long-term , chronic treatment [ 244805 ] . IDEC has signed an antibody humanization patent licensing agreement with Protein Design Labs [ 240591 ] . IDEC is also collaborating with Mitsubishi-Tokyo ( formerly Mitsubishi Kasei ) on the development of this antibody [ 178382 ] . DB01079 pharmacokinetics are similar in patients with severe renal insufficiency and in healthy subjects . DB01079 ( HTF 919 ) , a selective Q13639 receptor partial agonist with promotile activity throughout the gastrointestinal tract , is in development for the treatment of irritable bowel syndrome . In an open-label , parallel-group study , the pharmacokinetics of a single 12-mg oral dose of tegaserod in patients with severe renal insufficiency requiring hemodialysis were compared with data obtained from healthy subjects matched for age , weight , height , and gender ( n = 10 , both ) . The pharmacokinetics of tegaserod were similar in both groups ( AUC(0h-tz) , ng.h/ml : 14.6 +/- 8.5 vs. 14.3 +/- 7.1 ; Cmax , ng/ml : 4.6 +/- 2.3 vs. 5.1 +/- 2.2 ; tmax , h : 1.0 , for both ) . DB01079 had similar tolerability in renally impaired patients and healthy volunteers , with adverse events largely related to the gastrointestinal pharmacological actions of the drug . Therefore , no dose adjustment of tegaserod is necessary for patients with renal insufficiency .

MH_train_1167

MH_train_1167

MH_train_1167

interacts_with DB00619?

multiple_choice

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 ) . 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 . A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma . DB00004 , a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 ( P60568 ) , efficiently targets lymphoma cells expressing the high-affinity P60568 receptor ( IL-2R ) consisting of the alpha/p55/CD25 , beta/p75/CD122 , and gamma/ P31785 /CD132 chains . In vitro studies demonstrated that the retinoid X receptor ( RXR ) retinoid , bexarotene , at biologically relevant concentrations of 10(-6) M to 10(-8) M , upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox . To determine whether this biomodulatory effect could be recapitulated in vivo , we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene ( 75 mg/day-300 mg/day ) and denileukin diftitox ( 18 mcg/kg per day x 3 days every 21 days ) in a phase 1 trial . Overall response was 67 % ( 4 complete responses , 4 partial responses ) . Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day . Four patients experienced grade 2 or 3 leukopenia , and 2 had grade 4 lymphopenia . Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses ( 150 mg/day ) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells . [ Production of cytokines by peripheral blood mononuclear cells -- correlation with clinicomorphological features in laryngeal carcinoma ] . INTRODUCTION : In studied analyzed role of the cytokines in pathology of neoplasms of various origin the importance of these proteins in regulation of immunocompetent cells function has been described . The aim of this study was to estimate of cho sen cytokines concentration produced by peripheral blood mononuclear cells and in whole blood in patients with laryngeal carcinoma and to analyze the connection of cytokines profile with clinicopathological features . MATERIALA AND METHODS : 55 patients with squamous cell carcinoma of the larynx treated at ENT Department Medical University of Lodz between 2003-2007 were analyzed . For estimation of cytokine secretion the cultures of isolated peripheral blood mononuclear cells ( T lymphocytes ) and the whole blood were established . Production of cytokines in supernatants was detected by Elisa . Connections with clinicomorphological features ( pT , pN , Anneroth , Batsakis i Lunas ' classification ) were analyzed . RESULTS : Authors reported statistical correlation between chosen cytokines concentration and clinicomorphological parameters : pT and P60568 , P05231 , P10145 , TNFalpha produced by isolated cells and P60568 , P05231 , TNFa and IFNgamma in whole blood , pN and P10145 , P22301 , IFNgamma ; P00519 score and P05231 , TNFalpha , IFNgamma produced by isolated cells and P60568 , P05231 , P22301 , TNFalpha , IFNgamma in whole blood . CONCLUSION : Our studied indicated the important influence of proinflammatory and regulatory cytokines produced by immunocompetent cells for course of neoplasm disease , aggressiveness and advance in laryngeal carcinoma . Effect of isoniazid on glutathione biosynthesis and degradation in Mycobacterium smegmatis . The mechanism of glutathione ( DB00143 ) depletion by isoniazid ( DB00951 ) was studied in M. smegmatis . DB00951 increased the activity of gamma-glutamyl transferase ( P19440 ) whether added before medium inoculation or to actively growing cells . The activity of P19440 in cells grown from the beginning in DB00951 -containing medium increased significantly on growth days 2-6 . Three-day old M. smegmatis cells treated with DB00951 exhibited a 30-65 % increase in the activity of P19440 . The activities of gamma-glutamyl-cysteine synthase ( GGCS ) and DB00143 synthase ( GS ) were lowered by 50 and 56 % respectively on the second day of growth when M. smegmatis was grown in a medium supplemented with 1.5 mg DB00951 per L . In 3-day old M. smegmatis , DB00951 significantly inhibited the activities of DB00143 biosynthetic enzymes . The results demonstrate that the increased activity of P19440 and decreased activities of DB00143 biosynthetic enzymes are responsible for DB00143 depletion by DB00951 in M. smegmatis . Coinduction of endothelial nitric oxide synthase and arginine recycling enzymes in aorta of diabetic rats . Decreased availability of arginine and impaired production of NO ( nitric oxide ) have been implicated in the development of endothelial dysfunction . DB00155 formed by the NOS reaction is recycled to arginine by the citrulline-NO cycle , which is composed of NOS , argininosuccinate synthetase ( AS ) , and argininosuccinate lyase . Therefore , we investigated the alterations of these enzymes in the aorta of streptozotocin ( Q11206 ) -induced diabetic rats . P29474 and AS mRNAs were increased by three- to fourfold 1-2 weeks after Q11206 treatment and decreased at 4 weeks . AL mRNA was weakly induced . Induction of P29474 and AS proteins was also observed . Cationic amino acid transporter ( CAT ) -1 mRNA remained little changed , and CAT-2 mRNA was not detected . The plasma nitrogen oxide levels were increased 1-2 weeks after Q11206 treatment and decreased at 4 weeks . Transforming growth factor-beta1 ( TGF-beta1 ) mRNA in the aorta was also induced . TGF-beta1 induced P29474 and AS mRNAs in human umbilical vein endothelial cells but inhibited the proliferation of HUVEC . These results indicate that P29474 and AS are coinduced in the aorta in early stages of Q11206 -induced diabetic rats and that the induction is mediated by TGF-beta1 . The results also suggest that TGF-beta1 works antiatherogenically at early stages of diabetes by increasing NO production , whereas prolonged elevation of TGF-beta1 functions atherogenically by inhibiting endothelial cell growth . P20273 regulates adaptive and innate immune responses of B cells . B cells sense microenvironments through the B cell receptor ( P11274 ) and Toll-like receptors ( TLRs ) . While signals from P11274 and TLRs synergize to distinguish self from nonself , inappropriate regulation can result in development of autoimmune disease . Here we show that P20273 , an inhibitory co-receptor of P11274 , also negatively regulates TLR signaling in B cells . P20273 -deficient ( Cd22(-/-) ) B cells exhibit hyperactivation in response to ligands of TLRs 3 , 4 and 9 . Evidence suggests that this results from impaired induction of suppressors of cytokine signaling 1 and 3 , well-known suppressors of TLR signaling . Antibody-mediated sequestration of P20273 on wild-type ( WT ) B cells augments proliferation by TLR ligands . Conversely , expression of P20273 in a Cd22(-/-) B cell line blunts responses to TLR ligands . We also show that lipopolysaccharide-induced transcription by nuclear factor-κB is inhibited by ectopic expression of P20273 in a O00206 reporter cell line . Taken together , these results suggest that negative regulation of TLR signaling is an intrinsic property of P20273 . Since TLRs and P11274 activate B cells through different signaling pathways , and are differentially localized in B cells , P20273 exhibits a broader regulation of receptors that mediate adaptive and innate immune responses of B cells than previously recognized . PP2Cdelta ( Ppm1d , O15297 ) , an endogenous inhibitor of p38 MAPK , is regulated along with Trp53 and Cdkn2a following p38 MAPK inhibition during mouse preimplantation development . Preimplantation embryos utilize mitogen-activated protein kinase signaling ( MAPK ) pathways to relay signals from the external environment to prepare appropriate responses and adaptations to a changing milieu . It is therefore important to investigate how MAPK pathways are regulated during preimplantation development . This study was conducted to investigate whether PP2Cdelta ( Ppm1d , O15297 ) is expressed during mouse preimplantation development and to determine the influences of p38 MAPK inhibition on expression of Trp53 ( p53 ) , Ppm1d , ( O15297 ) , and Cdkn2a ( p16 ) during mouse preimplantation development . Our results indicate that Trp53 , Ppm1d , and Cdkn2a mRNAs and TRP53 and PP2Cdelta proteins are expressed throughout mouse preimplantation development . Treatment of 2-cell embryos with DB04338 ( potent inhibitor of p38 MAPK alpha/beta/ Q16539 /11 ) significantly increased Trp53 , Ppm1d and Cdkn2a and Mapk14 mRNA levels at 12 and 24 hr . Treatment of 8-cell embryos with DB04338 for 12 hr increased Trp53 , Ppm1d , and Cdkn2a mRNA levels , but not Mapk14 mRNA levels . Treatment of 8-cell embryos for 24 hr increased Trp53 , and Ppm1d mRNA levels , but decreased Cdkn2a and Mapk14 mRNA levels . Therefore , blockade of p38 MAPK activity is associated with embryo stage specific influences on Trp53 , Ppm1d , Cdkn2a , and Mapk14 expression during mouse preimplantation development . These results define downstream targets of p38 MAPK during preimplantation development and indicate that the p38 MAPK pathway regulates Trp53 , Ppm1d , and Cdkn2a expression . This study increases our understanding of the mechanisms controlling preimplantation development and of the interactions between preimplantation embryos and their culture environments . MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens . We propose the Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout ( MAGeCK ) method for prioritizing single-guide RNAs , genes and pathways in genome-scale CRISPR/Cas9 knockout screens . MAGeCK demonstrates better performance compared with existing methods , identifies both positively and negatively selected genes simultaneously , and reports robust results across different experimental conditions . Using public datasets , MAGeCK identified novel essential genes and pathways , including P00533 in vemurafenib-treated A375 cells harboring a P15056 mutation . MAGeCK also detected cell type-specific essential genes , including P11274 and P00519 , in KBM7 cells bearing a P11274 - P00519 fusion , and P08069 in HL-60 cells , which depends on the insulin signaling pathway for proliferation . Inhibition of c-kit receptor tyrosine kinase activity by DB00619 , a selective tyrosine kinase inhibitor . DB00619 ( formerly known as CGP 57148B ) is a known inhibitor of the c-abl , bcr-abl , and platelet-derived growth-factor receptor ( P09619 ) tyrosine kinases . This compound is being evaluated in clinical trials for the treatment of chronic myelogenous leukemia . We sought to extend the activity profile of DB00619 by testing its ability to inhibit the tyrosine kinase activity of c-kit , a receptor structurally similar to P09619 . We treated a c-kit expressing a human myeloid leukemia cell line , M-07e , with DB00619 before stimulation with Steel factor ( SLF ) . DB00619 inhibited c-kit autophosphorylation , activation of mitogen-activated protein ( Q96HU1 ) kinase , and activation of Akt without altering total protein levels of c-kit , Q96HU1 kinase , or Akt . The concentration that produced 50 % inhibition for these effects was approximately 100 nmol/L . DB00619 also significantly decreased SLF-dependent growth of M-07e cells in a dose-dependent manner and blocked the antiapoptotic activity of SLF . In contrast , the compound had no effect on Q96HU1 kinase activation or cellular proliferation in response to granulocyte-macrophage colony-stimulating factor . We also tested the activity of DB00619 in a human mast cell leukemia cell line ( HMC-1 ) , which has an activated mutant form of c-kit . DB00619 had a more potent inhibitory effect on the kinase activity of this mutant receptor than it did on ligand-dependent activation of the wild-type receptor . These findings show that DB00619 selectively inhibits c-kit tyrosine kinase activity and downstream activation of target proteins involved in cellular proliferation and survival . This compound may be useful in treating cancers associated with increased c-kit kinase activity . DB04630 stimulates vascular smooth muscle cell proliferation via big mitogen-activated protein kinase 1 activation . The nongenomic effects of aldosterone have been implicated in the pathogenesis of various cardiovascular diseases . DB04630 -induced nongenomic effects are attributable in part to the activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) , a classical mitogen-activated protein ( Q96HU1 ) kinase . Q13164 ( Q13164 ) , a newly identified Q96HU1 kinase , has been shown to be involved in cell proliferation , differentiation , and survival . We examined whether aldosterone stimulates Q13164 -mediated proliferation of cultured rat aortic smooth muscle cells ( RASMCs ) . P08235 ( MR ) expression and localization were evaluated by Western blotting analysis and fluorolabeling methods . P27361 /2 and Q13164 activities were measured by Western blotting analysis with the respective phosphospecific antibodies . Cell proliferation was determined by Alamar Blue colorimetric assay . DB04630 ( 0.1 to 100 nmol/L ) dose-dependently activated Q13164 in RASMCs , with a peak at 30 minutes . To clarify whether aldosterone-induced Q13164 activation is an MR-mediated phenomenon , we examined the effect of eplerenone , a selective MR antagonist , on aldosterone-induced Q13164 activation . DB00700 ( 0.1 to 10 micromol/L ) dose-dependently inhibited aldosterone-induced Q13164 activation in RASMCs . DB04630 also stimulated RASMC proliferation , which was inhibited by eplerenone . DB04630 -mediated phenomena were concluded to be attributable to a nongenomic effect because cycloheximide failed to inhibit aldosterone-induced Q13164 activation . Transfection of dominant-negative Q96HU1 kinase/ P29323 kinase 5 ( Q13163 ) , which is an upstream regulator of Q13164 , partially inhibited aldosterone-induced RASMC proliferation , which was almost completely inhibited by MEK inhibitor PD98059 . In addition to the classical steroid activity , rapid nongenomic effects induced by aldosterone may represent an alternative etiology for vascular diseases such as hypertension . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . [ 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 . The postnatal rat aorta contains pericyte progenitor cells that form spheroidal colonies in suspension culture . Pericytes play an important role in modulating angiogenesis , but the origin of these cells is poorly understood . To evaluate whether the mature vessel wall contains pericyte progenitor cells , nonendothelial mesenchymal cells isolated from the rat aorta were cultured in a serum-free medium optimized for stem cells . This method led to the isolation of anchorage-independent cells that proliferated slowly in suspension , forming spheroidal colonies . This process required basic fibroblast growth factor ( P09038 ) in the culture medium , because P09038 withdrawal caused the cells to attach to the culture dish and irreversibly lose their capacity to grow in suspension . Immunocytochemistry and RT-PCR analysis revealed the expression of the precursor cell markers P28906 and Tie-2 and the absence of endothelial cell markers ( CD31 and endothelial nitric oxide synthase , P29474 ) and smooth muscle cell markers ( alpha-smooth muscle actin , alpha-SMA ) . In addition , spheroid-forming cells were positive for Q99942 , nestin , PDGF receptor ( P09619 ) -alpha , and P09619 . Upon exposure to serum , these cells lost P28906 expression , acquired alpha-SMA , and attached to the culture dish . Returning these cells to serum-free medium failed to restore their original spheroid phenotype , suggesting terminal differentiation . When embedded in collagen gels , spheroid-forming cells rapidly migrated in response to DB00102 and became dendritic . Spheroid-forming cells cocultured in collagen with angiogenic outgrowths of rat aorta or isolated endothelial cells transformed into pericytes . These results demonstrate that the rat aorta contains primitive mesenchymal cells capable of pericyte differentiation . These immature cells may represent an important source of pericytes during angiogenesis in physiological and pathological processes . They may also provide a convenient supply of mural cells for vascular bioengineering applications . Molecular-based choice of cancer therapy : realities and expectations . Current choice of cancer therapy is usually empirical and relies mainly on the statistical prediction of the treatment success . Molecular research provides some opportunities to personalize antitumor treatment . For example , life-threatening toxic reactions can be avoided by the identification of subjects , who carry susceptible genotypes of drug-metabolizing genes ( e.g. P51580 , P22309 , P42898 , Q12882 ) . Tumor sensitivity can be predicted by molecular portraying of targets and other molecules associated with drug response . Tailoring of antiestrogen and trastuzumab therapy based on hormone and P04626 receptor status has already become a classical example of customized medicine . Other predictive markers have been identified both for cytotoxic and for targeted therapies , and include , for example , expression of TS , TP , Q12882 , OPRT , P07992 , P16455 , P11388 , class III beta-tubulin molecules as well as genomic alterations of P00533 , P10721 , P00519 oncogenes . A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors . P10721 or alpha-platelet-derived growth factor receptor ( alpha- P09619 ) activating mutations are the pathogenic mechanisms that characterize gastrointestinal stromal tumors ( GIST ) . Despite excellent responses to imatinib mesylate ( IM ) , patients are relapsing . We developed an IM-resistant GIST cell line ( GIST-R ) from the IM-sensitive GIST882 cell line ( GIST-S ) by growing these cells in IM . Gene expression profiling ( GEP ) of GIST-S , GIST-R cells and two IM resistant GIST patients demonstrated that P10721 is downregulated implying a major role in IM resistance . Instead , GIST-R cells have acquired IM resistance by overexpressing the oncogenic receptor tyrosine kinase - P30530 - in a ' kinase switch ' . Further , the two IM resistant GIST patients express P30530 and not c-Kit , seen by immunohistochemistry ( IHC ) . Real time reverse transcriptase-polymerase chain reaction and Western blotting of the GIST-S and GIST-R cells confirmed the switch from Kit to P30530 . In GIST-R , P30530 is tyrosine phosphorylated and its ligand growth-arrest-specific gene 6 is overexpressed implying autocrine activation . The kinase switch is associated with a morphological change from spindle to epithelioid . Molecular modeling of the kinase domain of mutant c-Kit ( V654A ) and P30530 showed no binding to IM but efficient binding to DB05216 , a novel c-Kit/ P30530 kinase inhibitor . DB05216 synergizes with docetaxel ( taxotere ) and is cytotoxic to GIST cells . Q9UBK8 Q9UBK8 66A > G has no effect on total homocysteine , folate , and DB00115 concentrations in renal transplant patients . The association of variants of the gene encoding methionine synthase reductase ( Q9UBK8 ) with hyperhomocysteinemia , folate and Vitamin B(12) status in kidney graft recipients is unknown . We examined two mutations in Q9UBK8 in a cross-sectional study of 733 kidney graft recipients . The allele frequency of Q9UBK8 66G was 0.55 . 369 patients ( 50.3 % ) were heterozygous and 219 patients ( 29.9 % ) were homozygous for the mutation . None of the patients showed the 997C > G mutation . The allelic variants of Q9UBK8 66A > G showed no significant association with total homocysteine ( tHcy ) levels , both in univariate analyses , and in a multivariate model controlling for age , gender , body mass index , renal function , time since transplantation , underlying kidney disease , as well as the P42898 677C > T/1298A > C genotypes . Similarly , no significant associations between the Q9UBK8 66A > Ggenotypes and plasma folate or Vitamin B(12) levels were found . In conclusion , Q9UBK8 66A > G has no major effect on tHcy , folate , or Vitamin B(12) plasma concentrations in kidney graft recipients . Targeting myeloid differentiation 2 for treatment of sepsis . Sepsis continues to be a leading cause of intensive care unit ( ICU ) death . Gram-negative bacteria are among the most important pathogens of sepsis and their LPS content is regarded to be an important stimulator that elicits the systemic inflammatory reaction . MD-2 is a small secreted glycoprotein that can bind to both the hydrophobic portion of LPS and to the extracellular domain of O00206 . The interaction between MD-2 and LPS bridges the two O00206 molecules and induces the dimerization of LPS-MD-2- O00206 , which forms the structural basis for biological functions of O00206 /MD-2 complex . Due to its essential role in mediating the interaction between LPS and O00206 , MD-2 has been extensively explored as a therapeutic target for treatment of inflammatory disorders such as sepsis . DB04933 is a synthetic tetraacylated lipid A that binds directly to MD-2 and antagonizes LPS binding to the same site . Although eritoran showed positive results in phase I and phase II clinical trials of severe sepsis , a phase III clinical study for severe sepsis has failed . More effective therapeutic strategies are in need to treat this devastating clinical disorder . Cloning and nucleotide sequence of human gamma-glutamyl transpeptidase . We have identified the gene for human gamma-glutamyl transpeptidase [ P19440 ; glutamine:D-glutamyl-peptide 5-glutamyltransferase ( also called gamma-glutamyltransferase ) , EC 2.3.2.2 ] in a P11274 gene-related region located in band q11 ---- qter of chromosome 22 . Two cDNAs complementary to the P19440 mRNA have been isolated from a human placental library constructed in phage lambda gt11 . The largest cDNA has a size of 2535 base pairs ( bp ) and an open reading frame of 1707 nucleotides encoding 569 amino acids . By using a probe corresponding to this cDNA , a mRNA of approximately 2.4 kilobases was detected by RNA blot-hybridization analysis in mouse kidney RNA . The P19440 precursor encoded by the coding sequence would have an estimated Mr of 61,400 . We compared our nucleotide and deduced amino acid sequences with the published results of rat kidney cDNAs . The human and rat amino acid sequences are similar ; however , a considerable discrepancy in nucleotide sequence was found within a 180-bp fragment of the heavy chain , resulting in a completely different amino acid sequence for this region . In addition , the 5' untranslated sequence of the human cDNA ( 669 bp ) is substantially larger than that determined in the rat cDNA ( 227 bp ) . Our results may be valuable for further studies on the protein structure of human P19440 as well as studies on the regulation of the enzyme .

MH_train_1168

MH_train_1168

MH_train_1168

interacts_with DB00338?

multiple_choice

Adolescent exposure to chronic DB00470 blocks opiate dependence in maternally deprived rats . Maternal deprivation in rats specifically leads to a vulnerability to opiate dependence . However , the impact of cannabis exposure during adolescence on this opiate vulnerability has not been investigated . Chronic dronabinol ( natural delta-9 tetrahydrocannabinol , THC ) exposure during postnatal days 35-49 was made in maternal deprived ( D ) or non-deprived ( animal facility rearing , AFR ) rats . The effects of dronabinol exposure were studied after 2 weeks of washout on the rewarding effects of morphine measured in the place preference and oral self-administration tests . The preproenkephalin ( PPE ) mRNA levels and the relative density and functionality of P21554 , and mu-opioid receptors were quantified in the striatum and the mesencephalon . Chronic dronabinol exposure in AFR rats induced an increase in sensitivity to morphine conditioning in the place preference paradigm together with a decrease of PPE mRNA levels in the nucleus accumbens and the caudate-putamen nucleus , without any modification for preference to oral morphine consumption . In contrast , dronabinol treatment on D-rats normalized PPE decrease in the striatum , morphine consumption , and suppressed sensitivity to morphine conditioning . P21554 and mu-opioid receptor density and functionality were not changed in the striatum and mesencephalon of all groups of rats . These results indicate THC potency to act as a homeostatic modifier that would worsen the reward effects of morphine on naive animals , but ameliorate the deficits in maternally D-rats . These findings point to the self-medication use of cannabis in subgroups of individuals subjected to adverse postnatal environment . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . 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 . 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 . Use of trifluoroperazine isolates a [(3)H] DB08954 binding site in rat brain membranes with the pharmacology of the voltage-independent ifenprodil site on N-methyl-D-aspartate receptors containing Q13224 subunits . The use of trifluoroperazine in a well washed rat brain membrane preparation revealed [(3)H]ifenprodil binding to a single high affinity state with the pharmacology of N-methyl-D-aspartate ( DB01221 ) receptors containing Q13224 subunits . Inhibition of [(3)H]ifenprodil binding in the presence of trifluoroperazine by 10 NR1a/ Q13224 selective agents was highly correlated with their inhibition at rat NR1a/ Q13224 receptors expressed in Xenopus ooctyes and [(3)H] DB01520 binding to rat brain Q13224 subunit containing DB01221 receptors but not with their inhibition of [(3)H]DTG binding . Allosteric interactions with polyamines , Mg(2+) , Zn(2+) , glutamate , glycine , and their antagonists were consistent with DB01221 receptors with Q13224 subtype pharmacology . The rank order of polyamine inhibition was spermine > spermidine > 1,5-(diethylamino)piperidine > arcaine > agmatine > putrescine . Both spermidine and MgCl(2) shifted the inhibition curve of ifenprodil to the right in a parallel manner , but Mg(2+) did not appear to be additive to spermidine . Glutamate increased and glycine decreased the binding . Conversely , CPP decreased the binding , and MDL 105,519 increased the binding in an agonist reversible manner . The increase with MDL 105,519 and glutamate appeared to be additive as did the decrease with glycine and CPP . Changes in the buffer pH between 6.5 and 8.0 did not affect the affinity of Q13224 agents . DB09202 but not clonidine inhibited the binding . MK-801 and agents from various other pharmacological classes did not significantly inhibit [(3)H]ifenprodil binding . [(3)H] DB08954 binding in the presence of trifluoroperazine appears to be selective for the voltage-independent ifenprodil site on DB01221 receptors containing the Q13224 subunit . Low calcium environment effects osteoprotegerin ligand/osteoclast differentiation factor . In coculture with osteoblastic cell line MC3T3-E1 ( E1 ) and mouse bone marrow cells , we reported that numbers of osteoclasts rose significantly on exposure to a low-calcium environment . Here we examined how osteoblasts influence osteoclastogenesis under a low-calcium environment . Comparing low extracellular calcium with a regular calcium environment , osteoprotegerin ligand ( O14788 ) /osteoclast differentiation factor ( O14788 ) mRNA expression show more increase in the culture of low-calcium environment than in that of a regular calcium environment . DB01373 -sensing receptor ( P41180 ) , which was supposed as one of the mechanisms of recognizing extracellular calcium , existedon the surface of E1 cells . When E1 cells stimulated with agonists of P41180 , gadolinium , and neomycin , O14788 / O14788 mRNA expression decreased . Moreover , these agonists reduced osteoclast formation in coculture . Taken together , it is possible that osteoblasts may recognize extracellular calcium via P41180 and regulate osteoclastogenesis . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity . Human cardiomyocyte hypertrophy induced in vitro by P40189 stimulation . OBJECTIVES : Recent in vivo and in vitro studies in animals have demonstrated that cytokines of the P05231 family are involved in cardiac hypertrophy and in protection of cardiomyocytes against apoptosis . The present study aims to analyse the capacity of human atrial cardiac cells ( i.e. , cardiomyocytes and fibroblasts ) to display the P40189 receptor subunit , and to evaluate its functionality . METHODS : Twenty human atrial biopsies were used for immunohistochemistry , in situ hybridisation , and western blot analysis or dissociated for isolation and primary culture of cardiac cells . RESULTS : Fibroblasts present in tissue or maintained in primary culture clearly express P40189 whereas the signal in cardiomyocytes is weaker . Culture of cardiac cells with a P40189 agonist antibody enhances atrial natriuretic peptide ( P01160 ) , beta myosin heavy chain ( beta-MHC ) expression in cardiomyocytes , and significantly increases the cell surface area microm(2) ) . This process could involve P40763 ( signal transducer and activator of transcription 3 ) phosphorylation . CONCLUSIONS : These results demonstrate that P40189 activation in human cardiac cells leads to cardiomyocyte hypertrophy . We discuss several hypotheses on the role of P05231 -type cytokines on cardiomyocyte functions . 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 . P10275 inducing bladder cancer progression by promoting an epithelial-mesenchymal transition . The study investigated the role of androgen receptor ( AR ) as a potential target for the treatment of bladder cancer in regulating epithelial-mesenchymal transition or transformation ( EMT ) . Cell proliferation , and migration capacity were determined in bladder cancer T24 cells treated with small interfering RNA directed against AR , and expression levels of P12830 , β-catenin and N- cadherin were assessed using quantitative reverse transcription PCR ( qRT-PCR ) . Tumour cell growth was evaluated in vivo in T24 tumour-bearing nude mice receiving electroporation-assisted administration of anti-AR small interfering RNA . It was found that low AR expression decreased proliferation and migration of bladder cancer cells . In vivo experiments showed that silencing AR expression significantly suppressed AR-positive bladder tumour growth with decreased cell proliferation . Low AR level of T24 bladder cancer cells treated with DB01541 ( DB02901 ) decreased expression of P12830 , β-catenin and P19022 expression , indicating a strong sensitivity to the EMT and In cells with low AR content , TGF-β induced down-regulation of P12830 and β-catenin . It is concluded that suppression of AR expression decreased the production of TGF-β , inhibiting EMT and bladder cancer cell growth in vitro and in vivo , implying that its use might be a potential therapeutic target for the treatment of bladder cancer . Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Cytokine signaling via various transcription factors regulates receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) -mediated osteoclast differentiation from monocyte/macrophage lineage cells involved in propagation and resolution of inflammatory bone destruction . Protein inhibitor of activated P40763 ( Q9Y6X2 ) was initially identified as a molecule that inhibits DNA binding of P40763 and regulates many transcription factors through distinct mechanisms . To analyze Q9Y6X2 function in osteoclasts in vivo , we have generated transgenic mice in which Q9Y6X2 is specifically expressed in the osteoclast lineage using the tartrate-resistant acid phosphatase ( TRAP ) gene promoter . Q9Y6X2 transgenic mice showed an osteopetrotic phenotype due to impairment of osteoclast differentiation . Overexpression of Q9Y6X2 in RAW264.7 cells suppressed O14788 -induced osteoclastogenesis by inhibiting the expression of c-Fos and O95644 . Interestingly , Q9Y6X2 inhibits the transcriptional activity of microphthalmia-associated transcription factor ( O75030 ) independent of sumoylation . Down-regulation of Q9Y6X2 markedly enhances O14788 -mediated osteoclastogenesis in RAW264.7 cells . Furthermore , overexpression of Q9Y6X2 in mouse primary osteoblast ( DB00925 ) , down-regulates O14788 expression induced by interleukin-6 ( P05231 ) cytokine family , and inhibits osteoclast formation from bone marrow macrophages ( BMMs ) in vitro coculture system . Down-regulation of Q9Y6X2 leads to the accelerated expression of O14788 in DB00925 stimulated with P05231 and soluble P05231 receptor ( sIL-6R ) . Taken together , our results clearly indicate that Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Loss of androgen receptor expression promotes a stem-like cell phenotype in prostate cancer through P40763 signaling . P10275 ( AR ) signaling is important for prostate cancer progression . However , androgen-deprivation and/or AR targeting-based therapies often lead to resistance . Here , we demonstrate that loss of AR expression results in P40763 activation in prostate cancer cells . AR downregulation further leads to development of prostate cancer stem-like cells ( CSC ) , which requires P40763 . In human prostate tumor tissues , elevated cancer stem-like cell markers coincide with those cells exhibiting high P40763 activity and low AR expression . AR downregulation-induced P40763 activation is mediated through increased interleukin ( IL ) -6 expression . Treating mice with soluble P05231 receptor fusion protein or silencing P40763 in tumor cells significantly reduced prostate tumor growth and CSCs . Together , these findings indicate an opposing role of AR and P40763 in prostate CSC development . Selective activation of the B natriuretic peptide receptor by P23582 ( P09543 ) . The natriuretic peptides are hormones that can stimulate natriuretic , diuretic , and vasorelaxant activity in vivo , presumably through the activation of two known cell surface receptor guanylyl cyclases ( P16066 and P20594 ) . Although atrial natriuretic peptide ( P01160 ) and , to a lesser extent , brain natriuretic peptide ( DB04899 ) are efficient activators of the P16066 guanylyl cyclase , neither hormone can significantly stimulate P20594 . A member of this hormone family , P23582 ( P09543 ) , potently and selectively activated the human P20594 guanylyl cyclase . P09543 does not increase guanosine 3',5'-monophosphate accumulation in cells expressing human P16066 . The affinity of P09543 for P20594 is 50- or 500-fold higher than P01160 or DB04899 , respectively . This ligand-receptor pair may be involved in the regulation of fluid homeostasis by the central nervous system . The Inhibition of Spinal Astrocytic O60674 - P40763 Pathway Activation Correlates with the Analgesic Effects of Triptolide in the Rat Neuropathic Pain Model . Neuropathic pain ( NP ) is an intractable clinical problem without satisfactory treatments . However , certain natural products have been revealed as effective therapeutic agents for the management of pain states . In this study , we used the spinal nerve ligation ( Q16658 ) pain model to investigate the antinociceptive effect of triptolide ( P28907 ) , a major active component of the traditional Chinese herb Tripterygium wilfordii Hook F . Intrathecal P28907 inhibited the mechanical nociceptive response induced by Q16658 without interfering with motor performance . Additionally , the anti-nociceptive effect of P28907 was associated with the inhibition of the activation of spinal astrocytes . Furthermore , intrathecal administration of P28907 attenuated Q16658 -induced janus kinase ( JAK ) signal transducers and activators of transcription 3 ( P40763 ) signalling pathway activation and inhibited the upregulation of proinflammatory cytokines , such as interleukin-6 , interleukin-1 beta , and tumour necrosis factor-α , in dorsal horn astrocytes . Moreover , Q13224 -containing spinal N-methyl D-aspartate receptor ( NMDAR ) was subsequently inhibited . Above all , P28907 can alleviate Q16658 -induced NP via inhibiting the neuroinflammation in the spinal dorsal horn . The anti-inflammation effect of P28907 may be related with the suppression of spinal astrocytic JAK- P40763 activation . Our results suggest that P28907 may be a promising drug for the treatment of NP . Characterization of the effect of chronic administration of a calcium-sensing receptor antagonist , DB05255 , on renal calcium excretion and serum calcium in postmenopausal women . Ronacaleret is an orally-active calcium-sensing receptor ( P41180 ) antagonist that has the potential for therapeutic utility in the stimulation of PTH release , notably as a bone anabolic agent comparable to recombinant human PTH(1-34) ( DB05829 (1-34) ) . A recent study has shown that , despite the ability to increase circulating PTH levels in postmenopausal women in a dose-dependent manner , minimal effects of DB05255 on bone mineral density have been observed . Therefore , the purpose of this study was to characterize the PTH profile as well as calcium metabolism parameters as a marker of PTH biological activity following the administration of DB05255 or DB05829 (1-34) . Administration of DB05255 led to lower peak levels of PTH than were observed with DB05829 (1-34) , however , greater total PTH exposure was observed . Further , chronic administration of either agent was associated with increases in urinary calcium excretion and serum calcium levels , with the magnitude of the changes following DB05255 significantly greater than that for DB05829 (1-34) . The greater magnitude of effects observed with DB05255 is likely due to the greater total PTH exposure , and is potentially reflective of a state comparable to mild hyperparathyroidism . It is not clear whether the administration of all calcilytics would lead to a similar result , or is due to characteristics specific to DB05255 . 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 . JAK/ P35610 pathways are not involved in the direct activation of antigen-presenting cells by contact sensitizers . JAK/ P35610 pathways are described as the major mechanisms by which cytokine receptors transduce intracellular signals . The signalling mechanisms in antigen-presenting cells ( P25054 ) in the sensitization phase of contact hypersensitivity are poorly understood . The aim of this study was to clarify whether well-established JAK/ P35610 signalling pathways might be activated directly by contact sensitizers as described previously for tyrosine kinases and some Q96HU1 kinases . As a model of epidermal P25054 , human monocytes and human monocyte-derived dendritic cells were stimulated with the structurally unrelated contact sensitizers D6RGH6 /MI , thimerosal , TNCB and formaldehyde . The phosphorylation states of the transcription factors P42224 , P40763 , Q14765 , P42229 and P42226 were determined by Western blot analysis using phosphospecific antibodies . In contrast to the positive controls performed with the cytokines P01579 , P22301 , IFN-alpha , GM- P04141 and P05112 , no significant increase in the phosphorylation of P35610 molecules was recognized in hapten-treated cells . These results suggest that contact allergens do not directly activate common JAK/ P35610 pathways . Therefore the activation of P25054 in the early sensitization phase of contact hypersensitivity by haptens does not involve signals normally delivered by JAK-associated cytokine receptors . Multifunctional specificity of the protein C/activated protein C Gla domain . DB00055 ( P25054 ) has potent anticoagulant and anti-inflammatory properties that are mediated in part by its interactions with its cofactor protein S and the endothelial cell protein C receptor ( Q9UNN8 ) . The protein C/ P25054 Gla domain is implicated in both interactions . We sought to identify how the protein C Gla domain enables specific protein-protein interactions in addition to its conserved role in phospholipid binding . The human prothrombin Gla domain , which can not bind Q9UNN8 or support protein S cofactor activity , has 22/45 residues that are not shared with the human protein C Gla domain . We hypothesized that the unique protein C/ P25054 Gla domain residues were responsible for mediating the specific interactions . To assess this , we generated 13 recombinant protein C/ P25054 variants incorporating the prothrombin residue substitutions . Despite anticoagulant activity similar to wild-type P25054 in the absence of protein S , P25054 variants P25054 (PT33-39) ( N33S/V34S/D35T/D36A/L38D/A39V ) and P25054 (PT36/38/39) ( D36A/L38D/A39V ) were not stimulated by protein S , whereas P25054 (PT35/36) ( D35T/D36A ) exhibited reduced protein S sensitivity . Moreover , PC(PT8/10) ( L8V/H10K ) displayed negligible Q9UNN8 affinity , despite normal binding to anionic phospholipid vesicles and factor Va proteolysis in the presence and absence of protein S . A single residue variant , PC(PT8) , also failed to bind Q9UNN8 . Factor VIIa , which also possesses DB00149 -8 , bound soluble Q9UNN8 with similar affinity to wild-type protein C , collectively confirming DB00149 -8 as the critical residue for Q9UNN8 recognition . These results reveal the specific Gla domain residues responsible for mediating protein C/ P25054 molecular recognition with both its cofactor and receptor and further illustrate the multifunctional potential of Gla domains . Effects of phenacetin and its metabolite p-phenetidine on P23219 and P35354 activities and expression in vitro . The present study was aimed to test the possible cyclooxygenase ( P36551 ) -1/ P35354 selectivity of the old analgesic drug phenacetin and its metabolite p-phenetidine , which exhibits high renal toxicity . DB00316 ( acetaminophen ) , the main metabolite of phenacetin with low renal toxicity , and indomethacin were selected as reference compounds . Collagen-stimulated platelet thromboxane B2 ( TxB2 ) production and phorbol 12-myristate-13-acetate ( PMA ) -induced neutrophil prostaglandin E2 ( DB00917 ) synthesis were used as indicators for P23219 and P35354 activity , respectively . DB03783 was even less potent than paracetamol to reduce the production of both TxB2 and DB00917 , and no clear preference for either of the P36551 -enzymes was seen . P-phenetidine was a more potent inhibitor , already at nanomolar level , of the synthesis of these prostanoids than indomethacin and showed some preference to P35354 inhibition . Somewhat higher , micromolar , concentrations of p-phenetidine also reduced P35354 expression in neutrophils . We suggest that the very potent inhibitory activity of p-phenetidine on DB00917 synthesis combined with the reduction of P35354 expression could explain the renal papillary necrosis in phenacetin kidney . Modulation of cytokine production and enhancement of cell viability by Q9NYK1 and Q9NR96 ligands during anthrax infection of macrophages . Inhalation of Bacillus anthracis , a bioterrorism agent , results in a high mortality rate despite appropriate antibiotic therapy . Macrophages appear to be a key factor in B. anthracis pathogenesis . The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax . However , preactivation of Toll-like receptors ( TLRs ) could modify the host response . TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens . We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli . We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines P05231 , P01375 , P22301 , and IL-12 p40 . Our studies showed that pretreatment of mouse macrophages with the Q9NR96 ligand DB05463 , or the Q9NYK1 ligands R-848 and IT-37 , results in a substantial decrease in the subsequent secretion of P05231 and P01375 in response to B. anthracis infection of macrophages . Furthermore , the Q9NYK1 and Q9NR96 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages . These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability .

MH_train_1169

MH_train_1169

MH_train_1169

interacts_with DB00605?

multiple_choice

Q07869 ( PPARalpha ) activators induce hepatic farnesyl diphosphate synthase gene expression in rodents . Fibrates are hypolipidemic drugs that exert multiple effects on lipid metabolism by activating peroxisome proliferator-activated receptor alpha ( PPARalpha ) and modulating the expression of many target genes . In order to investigate the link between PPARalpha and cholesterol synthesis , we analysed the effect of fibrates on expression of the farnesyl diphosphate synthase ( P14324 ) gene , known to be regulated by sterol regulatory element-binding proteins ( SREBPs ) , in conjunction with P04035 . In wild-type mice , both fenofibrate and WY 14,643 induced P14324 gene expression , an effect impaired in PPARalpha-null mice . A three-fold induction was observed in ciprofibrate-treated rat hepatocytes , in primary culture . This effect was decreased in presence of 5,6-dichlorobenzimidazole riboside ( DRB ) and cycloheximide ( CHX ) , transcription and translation inhibitors , respectively . Acyl- DB01992 oxidase ( Q15067 ) , a bona fide PPARalpha target gene , was induced by ciprofibrate but slower and more strongly than P14324 . In addition , induction of P14324 gene expression was abolished in the presence of 25-hydroxycholesterol ( 25-OH Chol ) . Thus , activation of PPARalpha by fibrates induced P14324 gene expression in both hepatocytes in culture and in mouse liver . This effect is likely to be dependent on cellular sterol level , possibly through SREBP-mediated transcriptional activation . Second-generation epidermal growth factor receptor tyrosine kinase inhibitors in lung cancers . P00533 mutations identify patients who are more likely to respond to treatment with epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors ( TKIs ) than cytotoxic chemotherapy . The distinct success of the first-generation P00533 TKIs erlotinib and gefitinib has been accompanied by the observation that acquired resistance to these treatments develops after a median of 1 year of treatment . Newer , second-generation P00533 TKIs have been developed with the intent to delay or overcome acquired resistance by the broader inhibition of kinases ( eg , P04626 and vascular endothelial growth factor receptor ) and/or altering the interactions with P00533 through irreversibly binding to the kinase domain . This article discusses many of these agents ( including afatinib , dacomitinib , DB05007 , AP26113 , and CO-1686 ) which have the potential for greater efficacy compared with first-generation P00533 TKIs , and may also have clinical activity against other oncogenic mutations within the P00533 family , including P04626 . Carbonic anhydrase inhibitors : DB00580 binds to a different active site region of the human isoform II as compared to the structurally related cyclooxygenase II " selective " inhibitor celecoxib . The high resolution X-ray crystal structure of the adduct of human carbonic anhydrase ( CA , EC 4.2.1.1 ) isoform II ( hCA II ) with the clinically used painkiller valdecoxib , acting as a potent CA II and cyclooxygenase-2 ( P35354 ) inhibitor , is reported . The ionized sulfonamide moiety of valdecoxib is coordinated to the catalytic Zn(II) ion with a tetrahedral geometry . The phenyl-isoxazole moiety of the inhibitor fills the active site channel and interacts with the side chains of Gln92 , Val121 , Leu198 , Thr200 , and Pro202 . Its 3-phenyl group is located into a hydrophobic pocket , simultaneously establishing van der Waals interactions with the aliphatic side chain of various hydrophobic residues ( Val135 , Ile91 , Val121 , Leu198 , and Leu141 ) and a strong offset face-to-face stacking interaction with the aromatic ring of Phe131 ( the chi1 angle of which is rotated about 90 degrees with respect to what was observed in the structure of the native enzyme and those of other sulfonamide complexes ) . Celecoxib , a structurally related P35354 inhibitor for which the X-ray crystal structure was reported earlier , binds in a completely different manner to hCA II as compared to valdecoxib . Celecoxib completely fills the entire CA II active site , with its trifluoromethyl group in the hydrophobic part of the active site and the p-tolyl moiety in the hydrophilic one , not establishing any interaction with Phe131 . In contrast to celecoxib , valdecoxib was rotated about 90 degrees around the chemical bond connecting the benzensulfonamide and the substituted isoxazole ring allowing for these multiple favorable interactions . These different binding modes allow for the further drug design of various CA inhibitors belonging to the benzenesulfonamide class . 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 . DB08911 . The mitogen-activated protein kinase ( MEK MAPK/ P29323 kinase ) signaling pathways play a critical role in regulation of diverse cellular activities , including survival , differentiation , proliferation , motility , and angiogenesis . Therefore , MEK inhibition was recognized as a promising target for antineoplastic therapy . While multiple MEK inhibitors have been tested clinically only trametinib ( GSK1120212 ) , an oral MEK inhibitor which is selective for Q02750 and P36507 has shown promising activity in several clinical trials on melanoma and colorectal cancer and it is being evaluated by the FDA for the treatment of metastatic melanoma . Mechanistically it was shown that trametinib induces cell cycle arrest in vitro . In this overview , important preclinical and clinical data for trametinib are presented including mechanism-based in vitro studies as well as findings from different clinical studies . Future clinical trial in different solid tumor entities will define the therapeutic role of this targeted therapy approach , possibly as a combination with other targeted therapies such as P15056 inhibitors . 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 . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Hsp27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp27 ( P04792 ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp27 decreased P05231 -dependent P40763 phosphorylation , nuclear translocation , and P40763 binding to the Twist promoter , suggesting that Hsp27 is required for P05231 -mediated EMT via modulation of P40763 /Twist signaling . We observed a correlation between Hsp27 and Twist in patients with prostate cancer , with Hsp27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp27 inhibition by DB06094 , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp27 as a critical regulator of P05231 -dependent and P05231 -independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer . A novel inhibitory mechanism of nitrogen-containing bisphosphonate on the activity of Cl- extrusion in osteoclasts . DB09152 -containing bisphosphonates have been well known to be inhibited farnesyl diphosphate synthase ( P14324 ) , an enzyme in mevalonic acid metabolism , resulting in disturbance in polymerization of cytoskeleton structure in bone resorption and promotion of apoptosis in mature osteoclasts . Although bisphosphonates have been reported to activate ion transporters in native epithelium and Xenopus oocytes , little is known whether bisphosphonates affect acid hydrochronic acid extrusion in osteoclasts during bone resorption . The aim of this study was to determine the role of bisphosphonates on inhibition of hydrochronic acid extrusion in osteoclasts . Effects of zoledronic acid , a nitrogen-containing bisphosphonate , on the Cl(-) current activated by extracellular acidification were examined in two types of osteoclasts derived from RAW264.7 cells and mouse bone marrow macrophages ( BMMs ) . Extracellular acidification induced outwardly rectifying Cl(-) currents in mouse osteoclasts . DB00399 dose-dependently inhibited the acid-activated Cl(-) current . The non-nitrogen bisphosphonate etidronic acid had no effect on the acid-activated Cl(-) current . DB00759 -induced P14324 silencing caused a significant decrease in the Cl(-) current . The inhibitor of geranylgeranyl transferase suppressed the Cl(-) current . By contrast , the inhibitory action of zoledronic acid was rescued by addition of geranylgeranyl acid , a derivative of mevalonic acid . The activity of acid-activated Cl(-) currents was dependent on expression of P51798 during osteoclastogenesis . These results suggest that nitrogen-containing bisphosphonates suppress the activity of osteoclastic acid-activated Cl(-) currents through P14324 inhibition , suggesting the inhibition of Cl(-) extrusion activity . Compound FLZ inhibits lipopolysaccharide-induced inflammatory effects via down-regulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways in RAW264.7 macrophages . AIM : The aim of this study was to investigate the effect of the squamosamide derivative FLZ ( N-2-(4-hydroxy-phenyl)-ethyl-2-(2,5-dimethoxy-phenyl)-3-(3-methoxy-4-hydroxy-phenyl)-acrylamide ) on lipopolysaccharide ( LPS ) -induced inflammatory mediator production and the underlying mechanism in RAW264.7 macrophages . METHODS : RAW264.7 cells were preincubated with non-toxic concentrations of compound FLZ ( 1 , 5 , and 10 micromol/L ) for 30 min and then stimulated with 10 microg/L LPS . The production of nitric oxide ( NO ) , the expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase 2 ( P35354 ) , and the activation of nuclear factor kappa-B ( NF-kappaB ) and mitogen-activated protein kinase ( MAPK ) pathways were examined . RESULTS : FLZ significantly inhibited the LPS-induced production of NO , as well as the expression of P35228 and P35354 at both the RNA and the protein levels in RAW264.7 cells . The LPS-induced increase in the DNA binding activity of NF-kappaB and activator protein 1 ( AP-1 ) , the nuclear translocation of NF-kappaB p65 , the degradation of the inhibitory kappaBalpha protein ( P25963 ) and the phosphorylation of P25963 , O15111 ( IKK ) alpha/beta , c-Jun NH(2)-terminal kinase ( JNK ) and p38 MAPKs were all suppressed by FLZ . However , the phosphorylation of extracellular signal-regulated kinase ( P29323 ) was not affected . Further study revealed that FLZ inhibited the phosphorylation of transforming growth factor-beta ( TGF-beta ) -activated kinase 1 ( TAK1 ) , which is an upstream signaling molecule required for IKKalpha/beta , JNK and p38 activation . CONCLUSION : FLZ inhibited the LPS-induced production of inflammatory mediators at least partly through the downregulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways . 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 . Cyclooxygenase 2-derived prostaglandin E2 production by corticotropin-releasing hormone contributes to the activated DB02527 response element binding protein content in rheumatoid arthritis synovial tissue . OBJECTIVE : To determine a mechanism by which corticotropin-releasing hormone ( P06850 ) promotes human inflammatory joint disease progression . METHODS : An ex vivo synovial tissue culture system was established to investigate the functional properties of P06850 at peripheral sites of inflammation . P06850 - and interleukin-1 beta ( P01584 ) -induced prostaglandin E(2) ( PGE(2) ) production from 10 fresh rheumatoid arthritis ( RA ) synovial tissue ( ST ) explants was quantified using a competitive enzyme-linked immunosorbent assay . Modulation of PGE(2) levels was further examined following selective and nonselective cyclooxygenase 2 ( P35354 ) inhibition . Nuclear extracts were analyzed by electrophoretic mobility shift assays to determine functional DB02527 response element binding protein ( CREB ) activity in response to P06850 and PGE(2) in isolated primary synovial cell populations . Western blot analysis measured levels of total and activated ( phosphospecific ) CREB/activating transcription factor ( P39905 ) family members prior to and following stimulation . RESULTS : P06850 , in a time- and dose-dependent manner , significantly ( P = 0.022 ) up-regulated PGE(2) production from 10 fresh RA ST explants . Costimulation of RA ST with P06850 and P01584 significantly augmented ( P = 0.036 ) the effects on PGE(2) production additively over 24 hours . We demonstrated that selective P35354 inhibitors prevent the induction of PGE(2) by both P06850 and P01584 . Further , we provided evidence that P06850 and PGE(2) signal through the induction of CREB and phosphorylated CREB/ P39905 family members in RA ST and in isolated primary RA cell populations . CONCLUSION : Our findings underscore the pathogenic role that P06850 may play in modulating inflammatory joint disease and establish the CREB/ P39905 family of transcription factors as principal effector molecules of proinflammatory mediator action in RA . 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 . Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 gene deletion and DB01997 were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety . P10275 repression of gonadotropin-releasing hormone gene transcription via enhancer 1 . DB00644 ( DB00644 ) plays a major role in the hypothalamic-pituitary-gonadal ( HPG ) axis , and synthesis and secretion of DB00644 are regulated by gonadal steroid hormones . Disruptions in androgen levels are involved in a number of reproductive defects , including hypogonadotropic hypogonadism and polycystic ovarian syndrome . Androgens down-regulate DB00644 mRNA synthesis in vivo and in vitro via an androgen receptor ( AR ) -dependent mechanism . DB02998 ( R1881 ) , a synthetic AR agonist , represses DB00644 expression through multiple sites in the proximal promoter . In this study , we show AR also represses DB00644 transcription via the major enhancer ( DB00644 -E1 ) . A multimer of the -1800/-1766 region was repressed by R1881 treatment . Mutation of two bases , -1792 and -1791 , resulted in decreased basal activity and a loss of AR-mediated repression . AR bound to the -1796/-1791 sequence in electrophoretic mobility shift assays , indicating a direct interaction with DNA or other transcription factors in this region . We conclude that AR repression of DB00644 -E1 acts via multiple AR-responsive regions , including the site at -1792/-1791 . P55157 inhibitor decreases plasma cholesterol levels in P01130 -deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 ) -inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) -2-cyclopentyl-2-[4-[(2,4-dimethyl-9H-pyrido[2,3-b]indol-9-yl)methyl]phenyl]-N- [ ( 1S ) -2-hydroxy-1-phenylethyl ] ethanamide ( DB04852 ) , a new P55157 inhibitor , to low-density lipoprotein ( LDL ) -receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg/kg for 4 weeks . In the 12 mg/kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E/LDL was not altered by DB04852 , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . 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 .

MH_train_1170

MH_train_1170

MH_train_1170

interacts_with DB00278?

multiple_choice

P08183 , Q9UNQ0 , and P60484 determine the response of glioblastoma to temozolomide and ABT-888 therapy . PURPOSE : Little is known about the optimal clinical use of ABT-888 ( veliparib ) for treatment of glioblastoma . ABT-888 is a PARP inhibitor undergoing extensive clinical evaluation in glioblastoma , because it may synergize with the standard-of-care temozolomide ( DB00853 ) . We have elucidated important factors controlling ABT-888 efficacy in glioblastoma . EXPERIMENTAL DESIGN : We used genetically engineered spontaneous glioblastoma mouse models and allograft models that were orthotopically transplanted into wild-type ( WT ) and Abcb1/Abcg2-deficient ( KO ) recipients . RESULTS : ABT-888/ DB00853 is not efficacious against p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR allografts in wild-type recipients , indicating inherent resistance . Abcb1/Abcg2 mediated efflux of ABT-888 at the blood-brain barrier ( BBB ) causes a 5-fold reduction of ABT-888 brain penetration ( P < 0.0001 ) that was fully reversible by elacridar . Efficacy studies in WT and KO recipients and/or concomitant elacridar demonstrate that Abcb1/Abcg2 at the BBB and in tumor cells impair DB00853 /ABT-888 combination treatment efficacy . DB04881 also markedly improved DB00853 /ABT-888 combination treatment in the spontaneous p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR glioblastoma model . Importantly , ABT-888 does enhance DB00853 efficacy in Pten deficient glioblastoma allografts and spontaneous tumors , even in Abcb1/Abcg2 proficient wild-type mice . Loss of P60484 occurs frequently in glioblastoma ( 36 % ) and in silico analysis on patient with glioblastoma samples revealed that it is associated with a worse overall survival ( 310 days vs. 620 days , n = 117 ) . CONCLUSIONS : The potential of ABT-888 in glioblastoma can best be demonstrated in patients with P60484 null tumors . Therefore , clinical trials with ABT-888 should evaluate these patients as a separate group . Importantly , inhibition of P08183 and Q9UNQ0 ( by elacridar ) may improve the efficacy of DB00853 /ABT-888 therapy in all glioblastoma patients . Novel treatments of GERD : focus on the lower esophageal sphincter . Up to 50 % of patients with gastroesophageal reflux disease ( GERD ) still suffer from GERD symptoms despite proton pump inhibitor ( PPI ) therapy , indicating a need for new treatments . The lower esophageal sphincter ( LES ) plays a crucial role in maintaining the mechanical barrier necessary for prevention of gastric reflux . Transient LES relaxation ( TLESR ) is an important factor behind the occurrence of reflux , and preclinical studies have identified a number of targets for pharmacologic modification of TLESR . However , only gamma-aminobutyric acid ( GABA ) type B receptor ( GABA(B) ) agonists and metabotropic glutamate receptor 5 ( P41594 ) modulators have shown positive proof of concept in the clinical setting . The P41594 negative allosteric modulator DB05070 improved symptoms in GERD patients , but was associated with central side effects such as dizziness . DB00181 , a GABA(B) receptor agonist , reduces the incidence of TLESR and improves GERD symptoms in both adult and pediatric GERD patients . However , the utility of baclofen is similarly limited by poor tolerability and recent research has focused on the development of GABA(B) receptor agonists with improved tolerability . DB05031 , a prodrug of R-baclofen , reduced the number of reflux episodes in a dose-ranging study and was similarly tolerated to placebo . AZD3355 and AZD9343 are GABA(B) receptor agonists with limited central nervous system activity that have been shown in preclinical studies to reduce the incidence of TLESR and decrease esophageal acid exposure ; data from clinical studies of these agents in GERD patients are awaited with interest . Agents that target TLESR activity may therefore offer a promising new add-on treatment for patients who suffer from GERD symptoms despite PPI therapy . Structural basis for P01133 receptor inhibition by the therapeutic antibody DB05774 . Therapeutic anticancer strategies that target and inactivate the epidermal growth factor receptor ( P00533 ) are under intense study in the clinic . Here we describe the mechanism of P00533 inhibition by an antibody drug DB05774 . DB05774 is a fully human antibody that has similar antitumor potency as the chimeric cetuximab/Erbitux and might represent a safer therapeutic alternative . We report the X-ray crystal structure of the Fab fragment of DB05774 ( Fab11F8 ) in complex with the entire extracellular region and with isolated domain III of P00533 . We compare this to our previous study of the cetuximab/ P00533 interaction . Fab11F8 interacts with a remarkably similar epitope , but through a completely different set of interactions . Both the similarities and differences in binding of these two antibodies have important implications for the development of inhibitors that could exploit this same mechanism of P00533 inhibition . P21554 receptor antagonists for the treatment of nicotine addiction . Tobacco smoking is the largest cause of avoidable death and disease in developed countries . It is now viewed as a complex bio-psycho-social problem for which effective pharmacological treatments are needed . DB00184 is considered to be the primary compound of tobacco smoke that establishes and maintains tobacco dependence . The addictive effect of nicotine is mediated by activation of the mesolimbic system and the release of dopamine in the nucleus accumbens . Recently , the existence of a specific functional interaction between nicotine and the endocannabinoid system has been reported . Co-administration of sub-threshold doses of a cannabinoid agonist and nicotine produces rewarding effects and chronic nicotine treatment increases endocannabinoid levels in limbic regions . The P21554 receptor plays a key role in this interaction . P21554 knockout mice are less sensitive to the motivational effects of nicotine although this depends on the experimental model . The selective P21554 antagonist , rimonabant ( SR141716 ) , reduces nicotine self-administration and nicotine-seeking behavior induced by conditioned cues in rats . DB06155 appears to reduce nicotine addiction by attenuating the hyperactivation of the endocannabinoid system and the mesolimbic dopaminergic neuronal pathway . DB06155 may be considered as a potential alternative to the current substitutive treatments of nicotine addiction and may offer a new hope for the treatment of smokers who wish to quit . Cerebrospinal fluid P04271 levels reflect symptoms of depression in patients with non-inflammatory neurological disorders . Recent findings document numerous interactions between neuronal and glial systems that likely play a role in the pathophysiology of depression . These findings suggest that glia-derived neurotrophic protein P04271 may play a significant role in developing depression . To test the relationship between P04271 and depressive symptoms we designed cross-sectional clinical study including P04271 serum and P04141 levels in neurological patients with non-inflammatory disorders ( NIND ) , who undergone cerebrospinal fluid assessment for diagnostic purposes . The present study was focused on psychometric testing of depression ( BDI-II ) , anxiety ( SAS ) and alexithymia ( TAS-20 ) , and neurochemical measure of cerebrospinal fluid ( P04141 ) and serum levels of P04271 in 40 NIND inpatients [ mean age 41.67 ] . The main result shows that P04271 in P04141 is significantly negatively correlated with BDI-II ( Spearman R=-0.51 , p < 0.0009 ) but not with SAS and TAS-20 . The finding indicates that decreased level of P04271 in P04141 is related to increased symptoms of depression in the NIND patients . Glial proliferation and metabotropic glutamate receptor expression in amyotrophic lateral sclerosis . Accumulating evidence indicates that alterations in glial activation and disturbances in glial glutamate metabolism may contribute to the pathogenesis of amyotrophic lateral sclerosis ( P35858 ) . Metabotropic glutamate receptors ( mGluRs ) are involved in glutamate homeostasis as well as in glial proliferation . Using in situ hybridization and immunohistochemistry we found a strong upregulation of group I and group II mGluR mRNA and protein in P35858 spinal cord as compared to controls ( P41594 > Q13255 > Q14416 /3 ) . In vitro , the mGluR group I agonist 3,5-dihydroxyphenylglycine induced proliferation in chick spinal cord astroglial cultures . Moreover , addition of cerebrospinal fluid ( P04141 ) from P35858 patients resulted in significantly higher proliferation rates than control P04141 . In both cases , the effect could be blocked by addition of the mGluR group I antagonist 1-aminoindan-1,5-dicarboxylic acid . Taken together , our data suggest that stimulation of glial mGluRs through mediators present in the P04141 may contribute to glial proliferation and astrogliosis in P35858 . Phosphatidylinositol 3 kinase/Akt signal relay cooperates with smad in bone morphogenetic protein-2-induced colony stimulating factor-1 ( P09603 ) expression and osteoclast differentiation . Murine spleen cells produce mature osteoclasts when cocultured with osteoblastic cells . P04141 ( P04141 ) -1 is the growth factor required for differentiating the monocyte-macrophage precursor cells into preosteoclasts . Bone morphogenic protein ( BMP ) signaling in osteoblasts regulates bone mass in mice , suggesting a role of BMP in osteoclastogenesis along with osteoblast activity . The intracellular signal transduction cross talk regulating the osteoblastic production of P09603 as a mechanism of BMP-induced osteoclastogenesis is described in this report . We have recently described the involvement of Smad 1/5 in P12643 -induced P09603 expression and osteoclast formation . In this study , using the pharmacological inhibitors and the adenovirus ( Ad ) vectors expressing dominant-negative ( DN ) phosphatidylinositol 3 kinase ( PI3K ) , the PI3K-signaling inhibitor , phosphatase and tensin homolog deleted in chromosome 10 ( P60484 ) or DN Akt kinase in the in vitro coculture assay , we show an essential role of the lipid kinase cascade in P12643 -mediated multinucleated osteoclast formation and P09603 mRNA expression , transcription , and secretion . Inhibition of PI3K/Akt signaling blocked the binding of Smads 1/5 to the P09603 BMP-responsive element present in the P09603 promoter , resulting in attenuation of Smad-dependent P09603 transcription . Furthermore , PI3K inhibition and DN Akt prevented association of the transcriptional coactivator , CREB ( DB02527 response element binding protein ) binding protein ( CBP ) , with Smads 1/5 . Together , these data for the first time demonstrate that PI3K-dependent Akt activation regulates P12643 -induced P09603 expression and provides a mechanism for osteoblastic cell-assisted osteoclast differentiation . [ Regulation of P04271 expression during long term potentiation ] . In this study , contributions of intracellular regulatory cascades in the induction of P04271 expression in rat hippocampal P00915 area during long term posttetanic potentiation ( LTP ) were estimated . The activation of transcription factor p53 ( positive regulator of P04271 transcription ) by nutlin-3 increased the basal content of P04271 mRNA up to 151 % of the control level , which was significantly lower than its content in tetanized slices ( 280 % ) . Therefore , p53 seems to be not unique transcription factor upregulating P04271 expression during LTP . The inhibitor of Ca2+/calmodulin-dependent kinases ( CaMKs ) KN-93 fully blocked the increase of P04271 mRNA after tetanization , while KN-92 ( inactive analogue of KN-93 ) was ineffective . The inhibitor of CaMKII and receptor tyrosine kinases DB02152 essentially suppressed P04271 expression during LTP , the inhibition of MAPK p38 or P51812 moderately decreased , and the inhibition of Q02750 did not influence P04271 mRNA content . Thus , CaMKs play a key role in the induction of P04271 expression during LTP . Crystal structure of phenylalanine ammonia lyase : multiple helix dipoles implicated in catalysis . The first three-dimensional structure of phenylalanine ammonia lyase ( Q9P2V4 ) has been determined at 2.1 A resolution for Q9P2V4 from Rhodosporidium toruloides . The enzyme is structurally similar to the mechanistically related histidine ammonia lyase ( P42357 ) , with Q9P2V4 having an additional approximately 160 residues extending from the common fold . We propose that catalysis ( including lowering the pK(a) of nonacidic P01024 of l-phenylalanine for an E1cb mechanism ) is potentially governed by dipole moments of seven alpha helices associated with the Q9P2V4 active site ( six positive poles and one negative pole ) . Cofactor 3,5-dihydro-5-methylidene-4H-imidazol-4-one ( Q9NP71 ) resides atop the positive poles of three helices , for increasing its electrophilicity . The helix dipoles appear fully compatible with a model of phenylalanine docked in the active site of Q9P2V4 having the first covalent bond formed between the amino group of substrate and the methylidene group of Q9NP71 : 12 highly conserved residues ( near the N termini of helices for enhancing function ) are poised to serve roles in substrate recognition , Q9NP71 activation , product separation , proton donation , or polarizing electrons from the phenyl ring of substrate for activation of P01024 ; and a highly conserved DB00117 residue ( near the C terminus of the one helix that directs its negative pole toward the active site to increase the residue 's basicity ) is positioned to act as a general base , abstracting the pro-S hydrogen from P01024 of substrate . A similar mechanism is proposed for P42357 , which has a similar disposition of seven alpha helices and similar active-site residues . The helix dipoles appear incompatible with a proposed mechanism that invokes a carbocation intermediate . Protein composition and activation markers in plasma collected by three apheresis procedures . BACKGROUND : Scientific and technical advances made in transfusion medicine sustain the need for more comprehensive understanding of the impact of collection procedures on the quality of plasma for fractionation and for transfusion . This prospective work evaluated protein composition and markers of activation in plasma donations collected with three different automatic collection procedures ( performed on Haemonetics machines ) , including a new procedure using a high-separation core-molded bowl . STUDY DESIGN AND METHODS : A total of 90 collection procedures have been performed from a population of 37 donors , under comprehensively standardized conditions . Plasma aliquots were taken from the plasma units within 30 minutes of the end of the collection procedures and immediately frozen at -70 degrees C . Content in an extended range of proteins and of markers of activation of the coagulation and fibrinolytic systems has been measured using standard in vitro testing methods . RESULTS : Plasma donations had normal mean total protein , IgG , IgM , and fibrinogen content . The mean levels in coagulation FV , FVII , FVIII , and P03951 and in antithrombin were above the standard international requirements . There was no sign of activation of the hemostasis system , as assessed by activated FVII , thrombin antithrombin complex , P00734 fragment 1+2 , and D-dimers . Activated complement component P01024 and P01031 were low . CONCLUSION : Data indicates the good and consistent protein composition of plasma obtained by those automatic apheresis procedures . In particular , the new high-separation core procedure yields a high-quality plasma meeting requirements for transfusion and fractionation . NSA9 , a human prothrombin kringle-2-derived peptide , acts as an inhibitor of kringle-2-induced activation in EOC2 microglia . In neurodegenerative diseases , such as Alzheimer 's and Parkinson 's , microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds . P00734 and kringle-2 increase levels of NO and the mRNA expression of P35228 , IL-1beta , and P01375 in microglial cells . In contrast , the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1beta , P01375 , and P05231 in LPS-activated EOC2 microglia . In this study , we investigated the anti-inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia . Treatment with 20-100 muM of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation . NO production induced by MAPKs and NF-kappaB was similarly reduced by inhibitors of P29323 ( PD98059 ) , p38 ( SB203580 ) , NF-kappaB ( DB06151 ) , and NSA9 . These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2 . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD . 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 . Molecular analysis of the effect of topical imiquimod treatment of HPV 2/27/57-induced common warts . Imiquimod is effective in the treatment of genital warts and clinical studies suggest activity against common warts as well . We have analyzed the effect of topical imiquimod on gene expression and virus load in human papilloma virus ( HPV ) 2/27/57-induced common warts . mRNA was extracted from keratinocyte culture , from normal skin , from three untreated common warts and from three common warts treated topically with 5 % imiquimod cream twice daily . Differential gene expression was demonstrated by RT-PCR and by cDNA microarray hybridization . We further analyzed viral DNA content in scales from three superficially pared imiquimod-treated warts by real-time PCR . Comparison of normal skin with wart tissue revealed that HPV 2/27/57 infection led to an induction of P05231 , P22301 and interferon-gamma inducible protein ( IP10 ) and to an up-regulation of TGF-beta . We could further detect expression of PCTAIRE-3 , Q93097 , frizzled-3 , notch-2 , notch-4 and P51587 in normal skin and common warts . Analysis of imiquimod-treated warts demonstrated that imiquimod enhanced P05231 expression and induced P10145 , GM- P04141 , P05109 and P06702 . It could also be shown that imiquimod led to an infiltration of wart tissue with macrophages and to a strong decrease of viral copy number in warts within 3 months of treatment . Our data thus provide molecular proof of principle for imiquimod treatment of cutaneous common warts . alpha-Thrombin inhibits DNA synthesis in rat hepatocytes but not in hepatoma cells by receptor activation and proteolysis . P00734 is a plasma protein , which after tissue injury is converted to alpha-thrombin and is mainly involved in blood clot formation . It has also been shown to have a mitogenic effect on primary endothelial cells , vascular smooth muscle cells , fibroblasts and some tumor cells , but is an inhibitor of rat hepatocyte DNA synthesis on fibronectin matrix in cell culture . We now report that prothrombin is converted to alpha-thrombin by primary cultures of normal adult rat hepatocytes and alpha-thrombin is also a potent inhibitor of hepatocytes DNA synthesis . In contrast , rat hepatoma cells cultured under similar conditions were resistant to alpha-thrombin mediated DNA synthesis inhibition . The inhibitory effect of alpha-thrombin on DNA synthesis was antagonized by hirudin and antithrombin , two specific alpha-thrombin inhibitors or by the presence of collagen-I matrix . A thrombin receptor activating peptide ( TRAP6 ) also inhibited P01133 -mediated rat hepatocyte DNA synthesis , suggesting a role of the thrombin receptors in this process . Matrix fibronectin was degraded by alpha-thrombin . However , no appreciable cell detachment was observed . These results suggest a role of alpha-thrombin as a potent growth inhibitor of normal hepatocytes , possibly through control of fibronectin or other matrix protein(s) . 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 . 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 . Inhibition of P51587 and DB01643 Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 mediates homologous recombination repair , and P51587 polymorphisms increase cancer risk . However , tumors with P51587 mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 synergistically potentiated drugs with mechanisms of action related to P51587 function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality. " TS knockdown induced complementary lethality to TS-targeting drugs ( 5-FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 -targeting antisense oligdeoxynucleotide ( ASO ) " BR-1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi:10.1038/mtna.2013.7 published online 12 March 2013 . 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 .

MH_train_1171

MH_train_1171

MH_train_1171

interacts_with DB01039?

multiple_choice

DB01404 as a potential novel addition to the antikeloid weaponry . Keloid scars are large protruding claw-shaped lesions that develop beyond the confines of the wound and uniquely appears only in humans . For thousands of years ginseng has been used in the traditional medicine in oriental countries . It occupies a prominent position in the list of the best-selling medicinal herbs in the world . Panax ginseng often called Asian or Korean ginseng , is the most extensively used and the best grade of ginseng and the term of ' ginseng ' generally refers to Panax ginseng . Previous studies have revealed that ginseng inhibits NF-kappa B , TGF-β , P05231 , P12821 and P08253 and these factors play a pivotal role in keloid formation pathogenesis . Therefore it could be reasoned that ginseng could be effective for the treatment of the keloid scars . Clinical studies by topical applications of iPanax notoginseng ( 800 µg/ml ) are warranted . 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 . Structure and function of eritadenine and its 3-deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d- DB03769 ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 ) and has hypocholesterolemic activity . We have hypothesized that 3-deaza-DEA ( P01024 -DEA ) and its analogues retain high level of P23526 inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 -DEA analogues would have much higher hypocholesterolemic activity . P01024 -DEA , and its methyl ester ( P01024 -OMeDEA ) and its methyl amido ( P01024 -NMeDEA ) were synthesized to examine their P23526 inhibitory and hypocholesterolemic activities . A crystal structure of P23526 containing P01024 -DEA was determined and confirmed that DEA and P01024 -DEA bound to the same site of P23526 with the same binding mode . The P23526 inhibitory activities of P01024 -DEA ( K(I)=1.5 microM ) and P01024 -OMeDEA ( K(I)=1.5 microM ) are significantly lower than that of DEA ( K(I)=30 nM ) , while rats fed by P01024 -DEA and P01024 -OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 -NMeDEA lost most of the P23526 inhibitory activity ( K(I)=30 microM ) and dietary P01024 -NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 -DEA analogues are neither substrates nor inhibitors of adenosine deaminase . 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 . Pomegranate flower extract diminishes cardiac fibrosis in Zucker diabetic fatty rats : modulation of cardiac endothelin-1 and nuclear factor-kappaB pathways . The diabetic heart shows increased fibrosis , which impairs cardiac function . Endothelin ( ET ) -1 and nuclear factor-kappaB ( NF-kappaB ) interactively regulate fibroblast growth . We have recently demonstrated that Punica granatum flower ( P49763 ) , a Unani anti-diabetic medicine , is a dual activator of peroxisome proliferator-activated receptor ( Q07869 ) -alpha and -gamma , and improves hyperglycemia , hyperlipidemia , and fatty heart in Zucker diabetic fatty ( ZDF ) rat , a genetic animal model of type 2 diabetes and obesity . Here , we demonstrated that six-week treatment with P49763 extract ( 500 mg/kg , p.o. ) in Zucker diabetic fatty rats reduced the ratios of van Gieson-stained interstitial collagen deposit area to total left ventricular area and perivascular collagen deposit areas to coronary artery media area in the heart . This was accompanied by suppression of overexpressed cardiac fibronectin and collagen I and III mRNAs . Punica granatum flower extract reduced the up-regulated cardiac mRNA expression of ET-1 , P25101 , inhibitor-kappaBbeta and c-jun , and normalized the down-regulated mRNA expression of inhibitor-kappaBalpha in Zucker diabetic fatty rats . In vitro , Punica granatum flower extract and its components oleanolic acid , ursolic acid , and gallic acid inhibited lipopolysaccharide-induced NF-kappaB activation in macrophages . Our findings indicate that Punica granatum flower extract diminishes cardiac fibrosis in Zucker diabetic fatty rats , at least in part , by modulating cardiac ET-1 and NF-kappaB signaling . Regulation of hepatic ApoC3 expression by P20142 -1β mediates hypolipidemic effect of nicotinic acid . Peroxisome proliferator-activated receptor ( Q07869 ) γ coactivator-1β ( P20142 -1β ) is a transcriptional coactivator that induces hypertriglyceridemia in response to dietary fats through activating hepatic lipogenesis and lipoprotein secretion . The expression of P20142 -1β is regulated by free fatty acids . Here we show that P20142 -1β regulates plasma triglyceride metabolism through stimulating apolipoprotein P01024 ( P02656 ) expression and elevating P02656 levels in circulation . Remarkably , liver-specific knockdown of P02656 significantly ameliorates P20142 -1β-induced hypertriglyceridemia in mice . Hepatic expression of P20142 -1β and P02656 is reduced in response to acute and chronic treatments with nicotinic acid , a widely prescribed drug for lowering plasma triglycerides . Adenoviral-mediated knockdown of P20142 -1β or P02656 in the liver recapitulates the hypolipidemic effect of nicotinic acid . Proteomic analysis of hepatic P20142 -1β transcriptional complex indicates that it stimulates P02656 expression through coactivating orphan nuclear receptor ERRα and recruiting chromatin-remodeling cofactors . Together , these studies identify P20142 -1β as an important regulator of the P02656 gene cluster and reveal a mechanism through which nicotinic acid achieves its therapeutic effects . Mitochondrial 2,4-dienoyl- DB01992 reductases in the rat : differential responses to clofibrate treatment . The effect of a peroxisome proliferator , clofibrate , on mitochondrial 2,4-dienoyl- DB01992 reductases was studied in rat liver . The specific activity of reductase measured with 2,4-hexadienoyl- DB01992 as the substrate increased 2.9-fold in the liver homogenate and 2.5-fold in the mitochondrial extract , whereas acyl- DB01992 oxidase activity increased 13-fold and delta 3 , delta 2-enoyl- DB01992 isomerase activity , 25-fold in the homogenate . Chromatography of the rat liver homogenate on hydroxylapatite , which separates the mitochondrial isoforms ( M(r) 120,000 and M(r) 60,000 ) showed that the M(r) 60,000 isoform increased 3.5-fold and the M(r) 120,000 isoform 6-fold . When the isoforms were assayed with 2,4-hexadienoyl- DB01992 and trans-2,cis-4,7,10,13,16,19-docosaheptaenoyl- DB01992 , the activity ratios of P13671 to C22 were 1.5-2.1 for the both isoforms isolated from livers of either control or clofibrate-treated rats . A quantitative immunological experiment with the antibody for the 120,000 reductase in the mitochondrial extracts showed a 6.9-fold increase in the signal , confirming the observation that this isoform is induced more than the other . The mRNA levels of reductase , isomerase , and peroxisomal multifunctional enzyme ( MFE ) were found to rise in a parallel manner when analyzed by in situ or slot hybridizations , which suggests that the increase was mediated by the same mechanism . Peroxisome proliferators have been shown to increase the mRNA levels of MFE by inducing peroxisome proliferator-activated receptor ( Q07869 ) -mediated expression of the corresponding gene . The short-chain delta 3 , delta 2-enoyl- DB01992 isomerase and the M(r) 120,000 reductase are exceptions among the mitochondrial beta-oxidation proteins , which usually show only a minor response to peroxisome proliferators . P34995 antagonist reduces blood-brain barrier leakage after cerebral ischemia . Disruption of the blood-brain barrier ( BBB ) after cerebral ischemia is considered to be the initial step in the development of brain injuries , and an increase in the tyrosine phosphorylation of the tight junctional protein occludin has been shown to cause an increase in BBB permeability . DB00917 ( DB00917 ) appears to be associated with both toxic and protective effects on neuronal survival in vitro . However , it remains to be determined whether the prostanoid EP1 receptor is involved in the disruption of the BBB after cerebral ischemia . So we examined the effect of a prostanoid EP1 receptor antagonist , SC51089 , on BBB leakage and tyrosine phosphorylation of occludin after cerebral ischemia . We demonstrated that SC51089 attenuated the increase in the tyrosine phosphorylation of occludin in isolated brain capillaries , which was coincident with a decrease in BBB leakage . These results suggest that the prostanoid EP1 receptor is involved in the tyrosine phosphorylation of occludin at tight junction , which may lead to disruption of the BBB and be linked to the development of cerebral infarctions . 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 . Bradykinin-stimulated cyclooxygenase activity stimulates vas deferens epithelial anion secretion in vitro in swine and humans . Epithelia lining the male reproductive duct modulate fertility by altering the luminal environment to which sperm are exposed . Although vas deferens epithelial cells reportedly express high levels of cyclooxygenases ( Ptgs ) , and activation of bradykinin ( BK ) receptors can lead to upregulation of PTGS activity in epididymal epithelia , it remains unknown whether BKs and/or PTGSs have any role in modulating epithelial ion transport across vas deferens epithelia . Porcine and human vas deferens epithelial cell primary cultures and the PVD9902 cell line responded to lysylbradykinin with an increase in short circuit current ( I SC ; indicating net anion secretion ) , an effect that was 60 % -93 % reduced by indomethacin . The BK effect was inhibited by the B2 receptor subtype ( P30411 ) antagonist HOE140 , whereas the B1 receptor subtype agonist des-Arg9-BK had no effect . P30411 immunoreactivity was documented in most epithelial cells composing the native epithelium and on Western blots derived from cultured cells . Gene expression analysis revealed that the P35354 transcript is 20 times more abundant than its P23219 counterpart in cultured porcine vas deferens epithelia and that P30411 mRNA is likewise highly expressed . Subsequent experiments revealed that prostaglandin E2 , 1-OH prostaglandin E1 ( prostaglandin E receptor 4 [ P35408 ] agonist ) and butaprost ( PTGER2 agonist ) increase I SC in a concentration-dependent manner , whereas sulprostone ( mixed P34995 and P43115 agonist ) produced no change in I SC . These results demonstrate that autacoids can affect epithelial cells to acutely modulate the luminal environment to which sperm are exposed in the vas deferens by enhancing PTGS activity , leading to the production of prostaglandins that act at P35408 and/or PTGER2 to induce or enhance anion secretion . Differential recruitment of coregulator proteins steroid receptor coactivator-1 and silencing mediator for retinoid and thyroid receptors to the estrogen receptor-estrogen response element by beta-estradiol and 4-hydroxytamoxifen in human breast cancer . P03372 ( ER ) -alpha and Q92731 function as transcription factors , and both interact with nuclear regulatory proteins to enhance or inhibit transcription . We hypothesized that coregulators are expressed in breast cancer and may be differentially recruited by ERs in the presence of estrogen and tamoxifen . Q92731 was found to be expressed more frequently in node-negative patients ( P < 0.05 ) . Expression of steroid receptor coactivator-1 ( Q15788 ) was associated with nodal positivity ( P < 0.05 ) and resistance to endocrine treatment ( P < 0.001 ) . The spatial coexpression of P03372 , Q92731 , and the coregulatory proteins was established using immunofluorescence . In both cell lines ( MCF-7 and T47D ) and in primary breast cancer cell cultures , beta-estradiol up-regulated Q92731 and coregulator protein expression and increased P03372 / Q92731 interaction with the estrogen response element ( ERE ) . 4- Hydroxy-tamoxifen ( DB04468 ) increased P03372 and silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) expression and increased ER-ERE binding . Q15788 and Q9Y618 were identified at the ER-ERE complex , and interactions between ER isoforms and coregulatory proteins were determined using immunoprecipitation . Both P03372 and Q92731 preferentially bound Q15788 in the presence of beta-estradiol . Conversely , in cells treated with DB04468 , P03372 and Q92731 bound Q9Y618 . Differential recruitment of Q15788 and Q9Y618 by P03372 and Q92731 in the presence of beta-estradiol and DB04468 may be central to the response of the tumor to endocrine treatment . Changes in the levels of DNA methylation in testis and liver of SD rats neonatally exposed to DB01262 and cadmium . To investigate the changes in the levels of DNA methylation in the testis during development after neonatal transient exposure to DNA methyltransferase ( P26358 ) inhibitors , we orally administered Sprague-Dawley ( SD ) rats with DB01262 ( 5-aza-CdR ; 0.025 or 0.25 mg kg(-1) ) or cadmium ( Cd ; 1 , 2 or 4 mg kg(-1) ) daily from days 3-7 postpartum ( pp ) . Sperm numbers decreased at day 70 pp in all exposure groups . We then used a PCR-based assay , combined bisulfite restriction analysis ( COBRA ) and pyrosequencing to determine the degrees of DNA methylation . Both 5-aza-CdR and Cd reduced P26358 activity in vivo after 5 days ' exposure at day 8 pp but not at day 70 pp . In contrast , the DNA methylation level of LINE-1 was not changed in the testis , either at day 8 pp or at day 70 pp . We observed increased apoptosis and an increase in the p53 mRNA level , accompanied by a decreased DNA methylation level in the p53 gene promoter region , at day 8 pp in testis for the 5-aza-CdR-exposed groups but not in the Cd-exposed group . The Cd-exposed group exhibited a degradation of seminiferous tubules and inhibition of a stepwise change in methylation in the coding region of c-fos in testis at day 70 pp . Because we observed toxic phenotype development accompanied by aberrant DNA methylation , DNA methylation may play a role in chemical induced testis damage , with different P26358 inhibitors affecting DNA methylation levels in gene- or stage-specific manner . 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 . DB05767 ( Andrographis paniculata extract ) prevents development of murine colitis by inhibiting T-cell proliferation and Q8IXH7 /TH17 responses . BACKGROUND : Extracts of the plant Andrographis paniculata have been used to treat inflammatory diseases in Asian countries . A recent double-blind , placebo-controlled trial of DB05767 ( A. paniculata extract ) has demonstrated its safety and effectiveness for induction of clinical response , remission , and mucosal healing in patients with mild to moderate ulcerative colitis ( UC ) . We aimed to determine if DB05767 could prevent the development of T-cell-dependent murine colitis and to define its in vivo mechanism(s) of action . METHODS : CD(+)4CD45RB(high) T cells were transferred into Rag1(-/-) mice and gavaged daily with DB05767 or methyl cellulose ( MC ) . Severity of colitis was evaluated by weight loss , histology , and cytokine expression . RESULTS : Mice treated with MC developed colitis within 4-7 weeks , as evaluated by weight loss , and severe intestinal inflammation . DB05767 -treated mice did not lose weight and displayed only very mild intestinal inflammation . P01375 alpha ( P01375 -α ) , interleukin ( IL ) -1β , interferon-gamma ( IFN-γ ) , and Q9GZX6 expression were significantly decreased in DB05767 -treated mice . We observed higher percentages of naïve P01730 (+) T cells in the lamina propria of DB05767 -treated mice . At early timepoints DB05767 -treated mice have significantly reduced splenic cell counts , reduced P01730 (+) , and Q16552 (+) , and IFN-γ T(+) cells . Furthermore , DB05767 inhibited the proliferation of P01730 T cells and differentiation into Q8IXH7 /TH17 cells in vitro . CONCLUSIONS : DB05767 inhibits the development of chronic colitis by affecting early T-cell proliferation , differentiation , and TH(1)/TH(17) responses in a T-cell-driven model of colitis , presenting a unique mechanism of action . Our data suggest that DB05767 could be an attractive herbal therapeutic for inflammatory bowel disease . 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 . Trichloroethylene-induced gene expression and DNA methylation changes in B6C3F1 mouse liver . Trichloroethylene ( TCE ) , widely used as an organic solvent in the industry , is a common contaminant in air , soil , and water . Chronic TCE exposure induced hepatocellular carcinoma in mice , and occupational exposure in humans was suggested to be associated with liver cancer . To understand the role of non-genotoxic mechanism(s) for TCE action , we examined the gene expression and DNA methylation changes in the liver of B6C3F1 mice orally administered with TCE ( 0 , 100 , 500 and 1000 mg/kg b.w. per day ) for 5 days . After 5 days TCE treatment at a dose level of 1000 mg/kg b.w. , a total of 431 differentially expressed genes were identified in mouse liver by microarray , of which 291 were up-regulated and 140 down-regulated . The expression changed genes were involved in key signal pathways including Q07869 , proliferation , apoptosis and homologous recombination . Notably , the expression level of a number of vital genes involved in the regulation of DNA methylation , such as Utrf1 , Tet2 , P26358 , DNMT3a and DNMT3b , were dysregulated . Although global DNA methylation change was not detected in the liver of mice exposed to TCE , the promoter regions of Cdkn1a and Ihh were found to be hypo- and hypermethylated respectively , which correlated negatively with their mRNA expression changes . Furthermore , the gene expression and DNA methylation changes induced by TCE were dose dependent . The overall data indicate that TCE exposure leads to aberrant DNA methylation changes , which might alter the expression of genes involved in the TCE-induced liver tumorgenesis . 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 . Endothelin receptor antagonists as disease modifiers in systemic sclerosis . Systemic sclerosis ( SSc ) is a multisystem connective tissue disease of unknown etiology that is characterized by inflammation , vascular dysfunction and fibrosis of the skin and visceral organs . SSc is clinically diverse both in terms of the burden of skin and organ involvement and the rate of progression of the disease . Recent studies indicate that the endothelin system , especially ET-1 and the P25101 and ETB receptors may play a key role in the pathogenesis of SSc . A new class of drugs , endothelin receptor antagonists has been introduced for treatment of patients with pulmonary arterial hypertension ( PAH ) . DB00559 , a dual endothelin receptor antagonist as well as DB06268 and DB06403 , selective blockers of the P25101 receptor have proven effective in SSc-PAH . This effect may be mediated through both a vasodilatory and antifibrotic effect , thus making these agents attractive as potential disease modifying agents for SSc . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients .

MH_train_1172

MH_train_1172

MH_train_1172

interacts_with DB01037?

multiple_choice

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 . Homology models of the mutated P00533 and their response towards quinazoline analogues . One of the most intensely studied tyrosine kinases is the epidermal growth factor receptor ( P00533 ) . The tyrosine kinase receptors are known to be over expressed in some solid tumors and non-small cell lung cancers , causing differential susceptibility to the quinazoline inhibitors . In this study we have taken P43405 tyrosine kinase coordinates from PDB database to model two new P00533 receptors with these mutations G695S and L834R and conducted all the docking studies of the inhibitors , also evaluated these two models for quality of structure using PROCHECK . Seven quinazoline analogues ( gefitinib , erlotinib , DB05424 , and Q9Y259 -569 and other analogues ) were selected for comparisons among the two new models . This study determined the receptor/inhibitor interactions , at that active domain binding sites consisting of 15 amino acids . We were able to calculate the energy data for each of the seven inhibitors . This data has been important in interpreting the affinity between the inhibitors evaluated against the three models of P00533 ( wild-type and two mutated types ) . " Affinity " -based studies have indicated the order of response based on docking energy levels ( Van der Waals and electrostatic interactions ) . The active DB00171 binding sites consisting of 15 amino acid residues were identified and the total energy ( E(total) ) which showed the affinity between the inhibitor molecules and the receptor ( Van der Waals and electrostatic interactions ) . The selection of the quinazoline analogues was purely on their emergence as possible candidates in the drug discovery areas . This study describes the successful application of these models that we constructed for molecular docking studies to rationally design compounds predicted to bind favorably to the modeled P00533 catalytic sites . DB05229 sodium , a stable prostacyclin analogue , elicits dilation of isolated porcine retinal arterioles : roles of P29474 and potassium channels . PURPOSE : DB01240 ( DB01240 ) is usually described as an endoEDRFsthelium-derived relaxing factor , but the vasoreactivity to DB01240 in the retinal arterioles and the underlying mechanisms are not fully understood . We examined the effects of DB01240 on the retinal microcirculation using beraprost sodium ( BPS ) , a stable DB01240 analogue , and the signaling mechanisms involved in this vasomotor activity . METHODS : Porcine retinal arterioles were isolated , cannulated , and pressurized without flow in vitro . Video microscopic techniques recorded the diametric responses to BPS . RESULTS : DB05229 sodium elicited dose-dependent ( 0.1 pM-0.1 μM ) vasodilation of the retinal arterioles that was abolished by the P43119 ( IP ) antagonist CAY10441 . DB05229 sodium-induced vasodilation decreased by 50 % after the endothelium was removed and was inhibited by the nitric oxide ( NO ) synthase inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) comparable with denudation . Inhibition of soluble guanylyl cyclase by 1H-1,2,4-oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) and blockage of protein kinase A ( PKA ) by Rp-8-Br-cAMPS were comparable to L-NAME . DB05229 sodium-induced vasodilation was also inhibited by the nonselective potassium channel inhibitor , tetraethylammonium , and the adenosine triphosphate-sensitive potassium ( KATP ) channel blocker , glibenclamide . Residual vasodilation in the presence of glibenclamide decreased further with subsequent application of ODQ . CONCLUSIONS : DB05229 sodium , a stable DB01240 analogue , causes vasodilation of the retinal arterioles mediated via the IP receptor . The current findings suggest that BPS elicits endothelium-dependent and -independent dilation of the retinal arterioles mediated by NO induced by activation of PKA in the endothelium and the KATP channel activation in the vascular smooth muscle , respectively . Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals . In diabetic P01130 -/- mice , an ectopic P12643 -Msx2 gene regulatory program is upregulated in association with vascular calcification . We verified the procalcific actions of aortic Msx2 expression in vivo . CMV-Msx2 transgenic ( CMV-Msx2Tg(+) ) mice expressed 3-fold higher levels of aortic Msx2 than nontransgenic littermates . On high-fat diets , CMV-Msx2Tg(+) mice exhibited marked cardiovascular calcification involving aortic and coronary tunica media . This corresponded to regions of Msx2 immunoreactivity in adjacent adventitial myofibroblasts , suggesting a potential paracrine osteogenic signal . To better understand Msx2-regulated calcification , we studied actions in 10T1/2 cells . We found that conditioned media from Msx2-transduced 10T1/2 cells ( Msx2-CM ) is both pro-osteogenic and adipostatic ; these features are characteristic of Wnt signaling . Msx2-CM stimulated Wnt-dependent TCF/LEF transcription , and Msx2-transduced cells exhibited increased nuclear beta-catenin localization with concomitant alkaline phosphatase induction . Msx2 upregulated Wnt3a and Wnt7a but downregulated expression of the canonical inhibitor Dkk1 . Dkk1 treatment reversed osteogenic and adipostatic actions of Msx2 . DB06285 , a Q03431 agonist that inhibits murine vascular calcification , suppressed vascular P12643 -Msx2-Wnt signaling . Analyses of CMV-Msx2Tg(+) mice confirmed that Msx2 suppresses aortic Dkk1 and upregulates vascular Wnts ; moreover , TOPGAL(+) ( Wnt reporter ) ; CMV-Msx2Tg(+) mice exhibited augmented aortic LacZ expression . Thus , Msx2-expressing cells elaborated an osteogenic milieu that promotes vascular calcification in part via paracrine Wnt signals . Effect of conditioned media of acute myeloid leukemia blast cells on complement synthesis by cultured human cells of monocyte and hepatocyte origin . The effect of conditioned media of 3-day cultures of blast cells from peripheral blood of 5 patients with acute myeloid leukemia ( CM-AML ) was studied on the synthesis of P06681 , factor B ( Bf ) and P05155 ( DB05341 ) by human monocyte-macrophage cultures and HepG2 hepatoma cell line . The level of P06681 in the culture supernatants was measured by immune hemolysis , those of Bf and DB05341 by ELISA . CM-AML was added to the monocyte cultures on day 3 and replaced by culture fluid on day 6 . Compared to the control cultures , CM-AML significantly increased P06681 and Bf levels and slightly decreased DB05341 levels in the culture fluids on day 6 . On day 9 , Bf synthesis enhancement still could be observed but P06681 and DB05341 levels did not significantly differ from those of the control . CM-AML significantly increased the synthesis of factor B by the HepG2 cells too . A strong correlation was found between the results of the Bf protein and RNA determinations , which means that the supernatants of AML blasts affect the gene expression of factor B at a pretranslational level . The selective complement synthesis modifying effect of CM-AML was not due to interferons ( IFN ) because neither IFN-alpha nor P01579 could be detected in these conditioned media . The present findings indicate that the hypercomplementemia observed in AML patients can be due to unknown factor(s) produced by leukemic blast cells . [ DB09043 ( Eperzan ) : a new once-weekly agonist of glucagon-like peptide-1 receptors ] . DB09043 ( Eperzan ) is a new once-weekly agonist of Glucagon-Like Peptide-1 ( P0C6A0 ) receptors that is indicated in the treatment of type 2 diabetes . Two doses are available , 30 mg and 50 mg , to be injected subcutaneously once a week . It has been extensively evaluated in the HARMONY programme of eight large randomised controlled trials that were performed at different stages of type 2 diabetes , in comparison with placebo or an active comparator . The endocrine and metabolic effects of albiglutide are similar to those of other P43220 agonists : stimulation of insulin secretion ( incretin effect ) and inhibition of glucagon secretion , both in a glucose-dependent manner , retardation of gastric emptying and increase of satiety . These effects lead to a reduction in glycated haemoglobin ( HbA(1c) ) levels , combined with a weight reduction . The overall tolerance profile is good . DB09043 is currently reimbursed in Belgium after failure ( HbA(1c) > 7.5 % ) of and in combination with a dual therapy with metformin and a sulfonylurea as well as in combination with a basal insulin ( with or without oral antidiabetic drugs ) . To avoid hypoglycaemia , a reduction in the dose of sulfonylurea or insulin may be recommended . A once-weekly administration should increase patient 's acceptance of injectable therapy and improve compliance . 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 . Convergent and divergent cellular responses by ErbB4 isoforms in mammary epithelial cells . Associations of ErbB4 ( Q15303 / Q15303 ) , the fourth member of the P00533 family , with cancer are variable , possibly as a result of structural diversity of this receptor . There are multiple structural isoforms of Q15303 arising by alternative mRNA splicing , and a subset undergo proteolysis that releases membrane-anchored and soluble isoforms that associate with transcription factors and coregulators to modulate transcription . To compare the differential and common signaling activities of full-length ( FL ) and soluble intracellular isoforms of Q15303 , four JM-a isoforms ( FL and soluble intracellular domain ( ICD ) CYT-1 and CYT-2 ) were expressed in isogenic MCF10A cells and their biologic activities were analyzed . Both FL and ICD CYT-2 promoted cell proliferation and invasion , and CYT-1 suppressed cell growth . Transcriptional profiling revealed several new and underexplored Q15303 -regulated transcripts , including : proteases/protease inhibitors ( P08254 and P07093 ) , the YAP/Hippo pathway ( P29279 , O00622 , and P09486 ) , the mevalonate/cholesterol pathway ( P04035 , Q01581 , P01130 , and Q9UBM7 ) , and cytokines ( P10145 , P78556 , and P09341 ) . Many of these transcripts were subsequently validated in a luminal breast cancer cell line that normally expresses Q15303 . Furthermore , ChIP-seq experiments identified O75689 , P02649 , P09486 , P16949 , and Q05195 as novel molecular targets of Q15303 . These findings clarify the diverse biologic activities of Q15303 isoforms , and reveal new and divergent functions . IMPLICATIONS : ErbB4 as a regulator of Hippo and mevalonate pathways provides new insight into milk production and anabolic processes in normal mammary epithelia and cancer . 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 . P02649 genetic associations with seizure development after severe traumatic brain injury . PRIMARY OBJECTIVE : The purpose of the current study is to assess the role of the P02649 genotype in post-traumatic seizure ( Q03393 ) development . RESEARCH DESIGN : A retrospective study of 322 adult Caucasians with a severe TBI and P02649 genotype . METHODS AND PROCEDURES : Medical records were searched for Q03393 . Time to first seizure was categorized as early , late or delayed-onset Q03393 . Potential Q03393 associations by genotype , grouped genotype and allele were investigated . MAIN OUTCOME AND RESULTS : No statistically significant associations were found . However , two out of the four individuals ( 50 % ) with the E4/E4 genotype had late/delayed-onset Q03393 . Furthermore , none with a E2/E2 or E2/E4 genotype seized in the late periods . CONCLUSIONS : The results of this study may suggest 4/4 as a risk genotype for late/delayed onset Q03393 and a potential neuroprotective role of the E2 allele . However , this study did not definitively support a role for the P02649 genotype in Q03393 susceptibility and indicates that larger populations are needed to fully evaluate the potential impact of P02649 on Q03393 . Novel phenolic antioxidants as multifunctional inhibitors of inducible P19320 expression for use in atherosclerosis . A series of novel phenolic compounds has been discovered as potent inhibitors of P01375 -inducible expression of vascular cell adhesion molecule-1 ( P19320 ) with concurrent antioxidant and lipid-modulating properties . Optimization of these multifunctional agents led to the identification of 3a ( DB05399 ) as a clinical candidate with demonstrated efficacies in animal models of atherosclerosis and hyperlipidemia . 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 . 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 . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . DB03886 -dependent hyperphenylalaninemia due to deficiency of 6-pyruvoyl tetrahydropterin synthase . We describe the clinical , neurologic , and biochemical findings in 10 patients with 6-pyruvoyl tetrahydropterin synthase ( 6- Q03393 ) deficiency from seven families , all of whom originate from one large tribe in Saudi Arabia . This deficiency presents with severe , early onset of failure to thrive , neurologic deterioration , and morbidity and mortality secondary to repeated episodes of bronchopneumonia or cardiorespiratory abnormalities . The urinary pterin excretion pattern indicates deficient activity of 6- Q03393 , which has been confirmed by direct enzyme assay in red blood cells of three patients . We treated our patients with combined use of tetrahydrobiopterin 20 mg/kg/d , L-dihydroxyphenylalanine 15 mg/kg/d , carbidopa 3.75 mg/kg/d , and L-5-hydroxytryptophan 5 mg/kg/d . Neurologic findings improved significantly in all after 5 to 24 months . Although head circumference and weight returned to the lower limit of normal in four , height normalized only in one of seven patients . Despite an unrestricted diet during combined therapy , blood phenylalanine and urinary excretion of neopterin and biopterin returned to normal . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . Cluster analysis of risk factor genetic polymorphisms in Alzheimer 's disease . Multiple genetic variants may contribute to the risk of developing Alzheimer 's disease . We have analyzed polymorphisms in 9 genes to determine whether particular combinations would contribute to this risk . The genes were P02649 , LDLr , P01034 , P07339 , P01375 , P56817 , P10636 , Q8IWL8 , P29474 , and Q12800 . Three risk groups for the disease were identified . Risk group I was younger , was heterozygous for the P01034 ( GA ) , CTSD2936 ( AG ) , P01375 -308 ( AG ) genetic variants . Risk group II was older , was homozygous for the -427 P02649 promoter polymorphism ( TT ) , and heterozygous for the P10636 deletion and for the Q8IWL8 variant ( QR ) . Group III had both the youngest and oldest subjects , were heterozygous for the -863 ( AC ) and -1031 ( CT ) P01375 promoter polymorphisms . All three groups carried the P02649 4 allele and were heterozygous for both P56817 polymorphisms . The control groups were carriers of the P02649 3 allele and were homozygous for the P56817 genetic variants . 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 . Systematic meta-analyses and field synopsis of genetic association studies in schizophrenia : the SzGene database . In an effort to pinpoint potential genetic risk factors for schizophrenia , research groups worldwide have published over 1,000 genetic association studies with largely inconsistent results . To facilitate the interpretation of these findings , we have created a regularly updated online database of all published genetic association studies for schizophrenia ( ' SzGene ' ) . For all polymorphisms having genotype data available in at least four independent case-control samples , we systematically carried out random-effects meta-analyses using allelic contrasts . Across 118 meta-analyses , a total of 24 genetic variants in 16 different genes ( P02649 , P21964 , DAO , P21728 , P14416 , P21917 , Q96EV8 , P47870 , Q13224 , HP , P01584 , P42898 , O75051 , P31645 , P04637 and P17752 ) showed nominally significant effects with average summary odds ratios of approximately 1.23 . Seven of these variants had not been previously meta-analyzed . According to recently proposed criteria for the assessment of cumulative evidence in genetic association studies , four of the significant results can be characterized as showing ' strong ' epidemiological credibility . Our project represents the first comprehensive online resource for systematically synthesized and graded evidence of genetic association studies in schizophrenia . As such , it could serve as a model for field synopses of genetic associations in other common and genetically complex disorders . P43220 activation inhibits VLDL production and reverses hepatic steatosis by decreasing hepatic lipogenesis in high-fat-fed P02649 *3-Leiden mice . OBJECTIVE : In addition to improve glucose intolerance , recent studies suggest that glucagon-like peptide-1 ( P0C6A0 ) receptor agonism also decreases triglyceride ( TG ) levels . The aim of this study was to evaluate the effect of P43220 agonism on very-low-density lipoprotein ( VLDL ) -TG production and liver TG metabolism . EXPERIMENTAL APPROACH : The P0C6A0 peptide analogues CNTO3649 and exendin-4 were continuously administered subcutaneously to high fat diet-fed P02649 *3-Leiden transgenic mice . After 4 weeks , hepatic VLDL production , lipid content , and expression profiles of selected genes involved in lipid metabolism were determined . RESULTS : CNTO3649 and exendin-4 reduced fasting plasma glucose ( up to -30 % and -28 % respectively ) and insulin ( -43 % and -65 % respectively ) . In addition , these agents reduced VLDL-TG production ( -36 % and -54 % respectively ) and VLDL-apoB production ( -36 % and -43 % respectively ) , indicating reduced production of VLDL particles rather than reduced lipidation of apoB . Moreover , they markedly decreased hepatic content of TG ( -39 % and -55 % respectively ) , cholesterol ( -30 % and -55 % respectively ) , and phospholipids ( -23 % and -36 % respectively ) , accompanied by down-regulation of expression of genes involved in hepatic lipogenesis ( Srebp-1c , Fasn , Dgat1 ) and apoB synthesis ( Apob ) . CONCLUSION : P43220 agonism reduces VLDL production and hepatic steatosis in addition to an improvement of glycemic control . These data suggest that GLP-receptor agonists could reduce hepatic steatosis and ameliorate dyslipidemia in patients with type 2 diabetes mellitus .

MH_train_1173

MH_train_1173

MH_train_1173

interacts_with DB04871?

multiple_choice

Synthetic peptides interacting with the 67-kd laminin receptor can reduce retinal ischemia and inhibit hypoxia-induced retinal neovascularization . The high-affinity 67-kd laminin receptor ( P08865 ) is expressed by proliferating endothelial cells during retinal neovascularization . The role of P08865 has been further examined experimentally by administration of selective P08865 agonists and antagonists in a murine model of proliferative retinopathy . These synthetic P08865 ligands have been previously shown to stimulate or inhibit endothelial cell motility in vitro without any direct effect on proliferation . In the present study , a fluorescently labeled P08865 antagonist ( P01133 (33-42) ) was injected intraperitoneally into mice and its distribution in the retina was assessed by confocal scanning laser microscopy . Within 2 hours this peptide was localized to the retinal vasculature , including preretinal neovascular complexes , and a significant amount had crossed the blood retinal barrier . For up to 24 hours postinjection , the peptide was still present in the retinal vascular walls and , to a lesser extent , in the neural retina . Non-labeled P01133 (33-42) significantly inhibited pre-retinal neovascularization in comparison to controls treated with phosphate-buffered saline or scrambled peptide ( P < 0.0001 ) . The agonist peptide ( Lam beta 1(925-933) ) also significantly inhibited proliferative retinopathy ; however , it caused a concomitant reduction in retinal ischemia in this model by promoting significant revascularization of the central retina ( P < 0.001 ) . Thus , P08865 appears to be an important target receptor for the modulation of retinal neovascularization . Agonism of this receptor may be valuable in reducing the hypoxia-stimulated release of angiogenic growth factors which drives retinal angiogenesis . Discovery and structure-activity relationship of ( 1R ) -8-chloro-2,3,4,5-tetrahydro-1-methyl-1H-3-benzazepine ( DB04871 ) , a selective serotonin P28335 receptor agonist for the treatment of obesity . The synthesis and SAR of a novel 3-benzazepine series of P28335 agonists is described . Compound 7d ( lorcaserin , APD356 ) was identified as one of the more potent and selective compounds in vitro ( pEC50 values in functional assays measuring [(3)H]phosphoinositol turnover : P28335 = 8.1 ; 5- Q13049 = 6.8 ; P41595 = 6.1 ) and was potent in an acute in vivo rat food intake model upon oral administration ( ED50 at 6 h = 18 mg/kg ) . DB04871 was further characterized in a single-dose pharmacokinetic study in rat ( t1/2 = 3.7 h ; F = 86 % ) and a 28-day model of weight gain in growing Sprague-Dawley rat ( 8.5 % decrease in weight gain observed at 36 mg/kg b.i.d. ) . DB04871 was selected for further evaluation in clinical trials for the treatment of obesity . No activation of human pregnane X receptor by hyperforin-related phloroglucinols . The acylated phloroglucinol , hyperforin , the main active ingredient of St . John 's Wort , exerts antidepressant properties via indirect inhibition of serotonin reuptake by selectively activating the canonical transient receptor potential channel 6 ( Q9Y210 ) . DB01892 treatment can lead to drug-drug interactions due to potent activation of the nuclear receptor O75469 ( O75469 ) , a key transcriptional regulator of genes involved in drug metabolism and transport . It was previously shown that synthetic acylated phloroglucinol derivatives activate Q9Y210 with similar potency as hyperforin . However , their interaction potential with O75469 remained unknown . Here we investigated five synthetic Q9Y210 -activating phloroglucinol derivatives and four Q9Y210 -nonactivating compounds compared with hyperforin and rifampicin for their potential to activate O75469 in silico and in vitro . Computational O75469 pharmacophore modeling did not indicate potent agonist or antagonist interactions for the Q9Y210 -activating derivatives , whereas one of them was suggested by docking studies to show both agonist and antagonist interactions . DB01892 and rifampicin treatment of HepG2 cells cotransfected with human O75469 expression vector and a P08684 promoter-reporter construct resulted in potent O75469 -dependent induction , whereas all Q9Y210 -activating compounds failed to show any O75469 activation or to antagonize rifampicin-mediated P08684 promoter induction . DB01892 and rifampicin treatment of primary human hepatocytes resulted in highly correlated induction of O75469 target genes , whereas treatment with the phloroglucinol derivatives elicited moderate gene expression changes that were only weakly correlated with those of rifampicin and hyperforin treatment . These results show that Q9Y210 -activating phloroglucinols do not activate O75469 and should therefore be promising new candidates for further drug development . 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 . Immunohistochemical analysis of brain lesions using P04271 and glial fibrillary acidic protein antibodies in arundic acid- ( DB05343 ) treated stroke-prone spontaneously hypertensive rats . Stroke-prone spontaneously hypertensive rats ( SHRSP ) used as a model of essential hypertension cause a high incidence of brain stroke on the course of hypertension . Incidences and sizes of brain lesions are known to relate to the astrocyte activities . Therefore , relation between brain damage and the expression profile of the astrocytes was investigated with morphometric and immunohistochemical analyses using astrocyte marker antibodies of P04271 and glial fibrillary acidic protein ( P14136 ) with or without arundic acid administration , a suppressor on the activation of astrocytes . Arundic acid extended the average life span of SHRSP . An increase in brain tissue weight was inhibited concomitant with a lower rate of gliosis/hemosiderin deposit/scarring in brain lesions . P04271 - or P14136 -positive dot and filamentous structures were decreased in arundic acid-treated SHRSP , and this effect was most pronounced in the cerebral cortex , white matter , and pons , and less so in the hippocampus , diencephalon , midbrain , and cerebellum . Blood pressure decreased after administration of arundic acid in the high-dose group ( 100 mg/kg/day arundic acid ) , but not in the low-dose group ( 30 mg/kg/day ) . These data indicate that arundic acid can prevent hypertension-induced stroke , and may inhibit the enlargement of the stroke lesion by preventing the inflammatory changes caused by overproduction of the P04271 protein in the astrocytes . North central cancer treatment group ( NCCTG ) N0537 : phase II trial of P15692 -trap in patients with metastatic breast cancer previously treated with an anthracycline and/or a taxane . INTRODUCTION : Angiogenesis is an established target for the treatment of MBC . DB08885 ( P15692 - Q8NHU6 ) is a humanized fusion protein , which binds P15692 , P49765 , and Q07326 -1 and -2 . PATIENTS AND METHODS : A 2-stage phase II study with primary end points of confirmed tumor response and 6-month progression-free survival ( PFS ) . If either end point was promising after the initial 21 patients , an additional 20 patients would be enrolled . Measurable disease , < 2 previous chemotherapy treatments , previous anthracycline or taxane therapy , and Eastern Cooperative Oncology Group performance status of 0 or 1 were required . DB08885 was given at a dose of 4 mg/kg intravenous every 14 days . RESULTS : Twenty-one patients were enrolled ; 71 % had visceral disease , 57 % were estrogen receptor negative , 19 % had P04626 (+) disease with previous trastuzumab treatment , and 33 % had 2 previous chemotherapy regimens . Partial response rate was 4.8 % ( 95 % confidence interval [ CI ] , 0.1 % -23.8 % ) and 6-month PFS was 9.5 % ( 95 % CI , 1.2 % -30.4 % ) . Neither primary end point met efficacy goals and the study was terminated . A median of 3 cycles was given . Median PFS was 2.4 months . Common grade 3 or 4 adverse events were hypertension ( 33 % ) , fatigue ( 19 % ) , dyspnea ( 14 % ) , and headache ( 14 % ) . Two cases of severe left ventricular dysfunction were noted . CONCLUSIONS : DB08885 did not meet efficacy goals in patients previously treated with MBC . Toxicity was as expected for anti- P15692 therapy . [ Neuroprotective effect of epigallocatechin gallate on oxidative-stress-injured retinal cells ] . OBJECTIVE : To investigate the neuroprotective effect of epigallocatechin gallate(EGCG) , the main extract from green tea , on the oxidative-stress-injured retinal ganglion cells . METHODS : Rat retinal ganglion cells ( RGC-5 ) were cultured into 3 groups ( normal control ; H2O2 ; H2O2 + EGCG or Trolox or DB02690 ) . In-situ TUNEL was used to detect the apoptosis of the RGC-5 cells . Dihydroethidium ( DHE ) assay was used to observe the intracellular ROS generation . The activation of nuclear enzyme , P09874 was quantitatively detected by Western blot and the cell viability was measured by MT method . RESULTS : Hydrogen peroxide reduced RGC-5 cell viability in a time-concentration-dependent manner . The treatment of 500 micromol/L H2O2 for 24 hours reduced RGC-5 cell viability by about 50 % of control . Hydrogen peraoxide caused apoptosis of the RGC-5 cell , obviously increased intracellular ROS generation and up-regulated the P09874 expression . The pretreatment with EGCG was able to markedly reduce the number of apoptotic cells , attenuate intracellular ROS generation . Furthermore , MTT assay showed that the pretreatment with EGCG ( 50 micromol/L ) increased the most cell viability to 87 % of control , but pretreatment with Trolox ( 100 micromol/L ) and DB02690 ( 100 micromol/L , a P09874 inhibitor ) recovered the most cell viability to 62 % and 71 % of control respectively . CONCLUSION : EGCG is able to effectively protect retinal ganglion cell against oxidative-stressed injury and can be used as a very potential neuroprotective drug . Effects of conjugated linoleic acid supplementation and exercise on post-heparin lipoprotein lipase , butyrylcholinesterase , blood lipid profile and glucose metabolism in young men . This study was designed to investigate the effects of conjugated linoleic acid ( DB01211 ) supplementation and endurance exercise training-induced changes on post-heparin lipoprotein lipase ( PH- P06858 ) and butyrylcholinesterase ( BChE ) activities along with leptin , insulin and lipid levels in plasma by a randomized double blind experiment . Eighteen sedentary male volunteers were randomly divided into DB01211 and Placebo ( P98160 ) supplementation groups . Both groups underwent daily supplementation of either 3g DB01211 or 3g placebo for 30 days , respectively , and performed exercise on a bicycle ergometer 3 times per week for 30-40 min at 50 % VO2 peak workload . For plasma glucose , insulin and leptin levels and BChE activity fasting blood was used . For PH- P06858 measurements , blood was collected 15 min after 50 IU/kg iv heparin injection . In all groups , there is a statistically significant decrease in BChE ( p = 0.03 , p = 0.02 ) and leptin ( p = 0.002 ) , insulin and HOMA-IR levels ( p = 0.02 ) . Exercise with or without DB01211 supplementation decreased insulin levels and increased insulin sensitivity . PH- P06858 activity was increased significantly in both groups , displaying increased fatty acid mobilization . We conclude that though DB01211 supplementation and exercise can affect these parameters , DB01211 is not more effective than exercise alone . Hence , a prolonged supplementation regime may be more effective . Taken together in our small study group , our findings display that BChE is a potential marker for synthetic function of liver , fat metabolism , an obesity marker , a function long overlooked . Essential role of P48995 channels in cardiomyoblasts hypertrophy mediated by 5- Q13049 serotonin receptors . Serotonin ( 5-HT ) participates in the development of cardiac hypertrophy through 5-HT(2A) serotonin receptors . The hypertrophic growth of cardiomyoblasts induced by 5-HT(2A) receptors involves the activation of the Ca(2+) responsive calcineurin/NFAT pathway . However , the mechanism whereby NFAT is activated by 5-HT(2A) receptors remains indeterminate . In this study , we examined whether transient receptor potential canonical ( TRPC ) channels participate in NFAT activation and hypertrophic response triggered by 5-HT . We demonstrate that P48995 expression is upregulated in 5-HT-treated rat cardiomyoblasts whereas Q9Y210 is induced in a mouse model of heart hypertrophy . Moreover , P48995 knockdown by small interfering RNA inhibits NFAT activation and hypertrophic response mediated by 5-HT(2A) receptors . These findings provide new insights about a mechanistic basis for the activation of the calcineurin/NFAT pathway by 5-HT(2A) receptors and highlight the critical role of P48995 in the development of cardiac hypertrophy . Identification of putative serum glycoprotein biomarkers for human lung adenocarcinoma by multilectin affinity chromatography and LC-MS/MS . Glycoproteins in human serum play fundamental roles in many biological processes , and also have clinical value as biomarkers for disease progression and treatment . In this study , we isolated glycoproteins from the sera of three healthy individuals and three lung adenocarcinoma patients using multilectin affinity chromatography . The recovered glycoproteins were subjected to treatment with peptide-N-glycosidase F ( Q96IV0 F ) and in-gel digestion by trypsin . Tryptic peptides were analyzed by nano-LC coupled to P19957 -MS/MS and the MS/MS spectra were processed by Bioworks 3.2 and an in-house bioinformatics tool , ProtAn . Approximately 90 % of the proteins identified contained more than one potential glycosylation site . Comparison of the serum glycoproteome of healthy and adenocarcinoma individuals revealed 38 cancer-selective proteins . Among them , 60 % have previously been reported as low abundance proteins in human sera . We identified several cancer-selective proteins that have been previously characterized as potential indicators of lung cancer in serum or plasma , including haptoglobin ( HP ) , inter-alpha-trypsin inhibitor heavy chain 4 ( ITI-H4 ) , complement P01024 precursor , and leucine-rich alpha-2-glycoprotein . In addition , plasma kallikrein ( P03952 ) and inter-alpha-trypsin inhibitor heavy chain 3 ( ITI-H3 ) were identified as being potentially elevated in the lung cancer group , and were validated by Western blot analysis . Furthermore , approximately 18 kDa plasma kallirein protein fragment was detected at high levels in 25 out of 28 adenocarcinoma patients , while one of the eight normal individuals showed moderate positive . The results suggest that P03952 represents a potential candidate serum biomarker of lung cancer . Pharmacological characterization of mitogen-activated protein kinase activation by recombinant human P28335 , 5- Q13049 , and P41595 receptors . The type 2 serotonin ( 5-HT(2) ) receptor subfamily is known to couple to phosphoinositide hydrolysis ( PI ) and the subsequent mobilization of intracellular Ca(2+) , as well as the release of arachidonic acid ( AA ) . Less is known of 5-HT(2)-mediated activation of the mitogen-activated protein kinase ( MAPK ) or extracellular signal-regulated kinase ( P27361 /2 ) signaling . The present study measured the relative efficacies and potencies of 5-HT agonists to activate P28482 in non-neuronal cells expressing recombinant human 5-HT(2A) , 5-HT(2B) , and 5-HT(2C(ISV)) receptors . 5-HT agonists stimulated P28482 activity via all three 5-HT(2) subtypes . There were no meaningful differences in the potencies or relative efficacies of these agonists to affect P28482 activity vs. PI accumulation or Ca(2+) mobilization , suggesting that these pathways may be sequentially linked . Indeed , P28482 activity was very sensitive to PKC inhibition and calcium chelation and insensitive to tyrosine kinase and P19957 -kinase inhibition . 5-HT(2) receptors efficiently couple to MAPK activation via sequential PI hydrolysis , and Ca(2+) mobilization . This profile differs from reports of " agonist-directed trafficking of receptor stimulus " between PI/Ca(2+) and AA pathways activated by 5-HT(2) receptors . 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 . 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 . Menstrual cycle-dependent febrile episode mediated by sequence-specific repression of poly(ADP-ribose) polymerase-1 on the transcription of the human serotonin receptor 1A gene . The serotonin receptor 1A ( encoded by the P08908 gene ) plays a critical role in serotonergic transmission and was linked with many human diseases . A 33-year-old woman with rare menstrual cycle-dependent fever showed abnormal estrogen profile and responded well to the P08908 agonist buspirone , suggesting that her fevers were allied to estrogen-related P08908 deficiency . We identified an adenine deletion 480-bases upstream of the translation start site ( i.e. , -480delA ) of P08908 in this patient . To determine the underlying mechanism of -480delA-mediated P08908 deficiency , we first showed that P08908 -480 region can be bound by multiple nuclear protein(s) . We then identified poly(ADP-ribose) polymerase ( P09874 ) as one of the proteins that binds to P08908 -480 region . Using P09874 overexpression and knockdown , our data demonstrated that P09874 represses P08908 transcription . Furthermore , P08908 -480delA promoter possesses increased interaction with P09874 and caused an additional reduction in transcription . Finally , 17β-estradiol administration further reduced transcription associated with the mutant promoter . Altogether , these data suggest that estrogen-induced hyperactivity of P08908 mutant promoter causes the reduction of P08908 mRNA and leads to the disruption of P08908 -mediate hypothermic regulation . This is the first report of P08908 mutation underlying menstrual cycle-dependent febrile episodes , and implies that similar " febrile episode " cases may also result from the dysfunction of serotonin transmission . The role of serotonin receptor subtypes in the behavioural effects of neuroleptic drugs . A paw test study in rats . The present study was designed to evaluate the roles of serotonin P08908 and 5-HT2 receptors in the effects of neuroleptic drugs in the paw test . This behavioural test has been shown to model both the antipsychotic efficacy as well as the extrapyramidal side-effect liability of neuroleptic drugs . Whereas the P08908 receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OHDPAT ) reduced the effects of the classical neuroleptic haloperidol , it increased the effects of the atypical neuroleptic clozapine . The 5-HT2 receptor antagonist ketanserin as well as the P28335 /5-HT2 receptor antagonist ritanserin , on the other hand reduced the effects of haloperidol , whereas the P28335 /5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ( DOI ) reduced the effects of clozapine . The most important finding , however , was that the behavioural effects of different ( putative ) neuroleptics ( fluphenazine , P35240 -39166 , remoxipride , prothipendyl , thioridazine and risperidone ) were differentially influenced by both 8-OHDPAT and DOI , suggesting that there are important differences between the neuronal mechanisms underlying the behavioural effects of these neuroleptic drugs , even within the subclasses of classical and atypical neuroleptics . Mechanism of oral absorbent DB05269 in lipid abnormalities in experimental uremic rats . BACKGROUND : We have reported that oral sorbent DB05269 ( Q9NRA2 ) is effective in delaying the induction of dialysis in patients with chronic renal failure ( CRF ) because of its effect on lipid metabolism . To clarify the precise mechanism of Q9NRA2 in lipid abnormalities in CRF , we examined the effect of Q9NRA2 on plasma lipid profile , total bile acids ( TBA ) , and lipoprotein lipase ( P06858 ) activity in experimental uremic rats . METHODS : Uremic rats were prepared using male Wistar rats by ligating 5/6 of the renal artery . Uremic rats were randomly divided into two groups as follows : a control group in which rats were maintained on the standard diet and an Q9NRA2 group in which rats were maintained on a diet containing 5 g of Q9NRA2 per 100 g of standard diet for 10 weeks . Plasma P06858 activity was measured as free fatty acid ( FFA ) generation after intravenous administration of heparin . RESULTS : Plasma creatinine at 1.5 +/- 0.1 mg/dl was lower in the Q9NRA2 group than the 1.9 +/- 0.5 mg/ml level in the control group . Q9NRA2 significantly decreased plasma total cholesterol from 192 +/- 29 to 142 +/- 25 mg/dl , triglycerides from 198 +/- 71 to 99 +/- 38 mg/dl , and TBA from 19.6 +/- 2.6 mumol/liter to 8.8 +/- 3.5 mumol/ml . Plasma P06858 activity at 0.22 +/- 0.01 mumol FFA/min/hr was significantly higher in the Q9NRA2 group than 0.15 +/- 0.03 mumol FFA/min/hr in the control group . CONCLUSIONS : These results suggest that Q9NRA2 may improve plasma lipid abnormalities by binding to bile acids in the intestinal lumen and preventing their reabsorption and inhibiting the reduction of P06858 activity in experimental uremic rats . 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 . Risk variants in the P04271 gene predict elevated P04271 serum concentrations in healthy individuals . Several lines of evidence suggest an important role of the P04271 protein and its coding gene in different neuropathological and psychiatric disorders like dementia , bipolar affective disorders and schizophrenia . To clarify whether a direct link exists between gene and gene product , that is , whether P04271 variants directly modulate P04271 serum concentration , 196 healthy individuals were assessed for P04271 serum concentrations and genotyped for five potentially functional P04271 SNPs . Functional variants of the serotonergic genes P08908 and 5-HTT possibly modulating P04271 serum levels were also studied . Further , publicly available human postmortem gene expression data were re-analyzed to elucidate the impact of P04271 , P08908 and 5-HTT SNPs on frontal cortex P04271 mRNA expression . Several P04271 SNPs , particularly rs9722 , and the P04271 haplotype T-G-G-A ( including rs2186358-rs11542311-rs2300403-rs9722 ) were associated with elevated P04271 serum concentrations ( Bonferroni corrected P < 0.05 ) . Of these , rs11542311 was also associated with P04271 mRNA expression directly ( Bonferroni corrected P = 0.05 ) and within haplotype G-A-T-C ( rs11542311-rs2839356-rs9984765-rs881827 ; P = 0.004 ) , again with the G-allele increasing P04271 expression . Our results suggest an important role of P04271 SNPs on P04271 serum concentrations and P04271 mRNA expression . It hereby links recent evidence for both , the impact of P04271 gene variation on various neurological or psychiatric disorders like dementia , bipolar affective disorders and schizophrenia and the strong relation between P04271 serum levels and these disorders . Contribution of the 37-kDa laminin receptor precursor in the anti-metastatic P08118 -derived peptide DB04985 cell surface binding . PURPOSE : DB04985 is an anti-metastatic synthetic peptide with promising therapeutic efficacy against hormone-refractory prostate cancer . The characterization of the DB04985 peptide cell surface binding/internalization mechanisms and of the receptors involved remained to be explored . RESULTS : [(14)C] DB04985 cell surface binding assays showed rapid and transient kinetic profile , that was inhibited by RGD peptides , laminin , hyaluronan , and type-I collagen . RGD peptides were however unable to inhibit DB04985 intracellular uptake . Far-Western ligand binding studies enabled the identification of the 37-kDa laminin receptor precursor ( P08865 ) as a potential ligand for DB04985 . Overexpression of the recombinant P08865 indeed led to an increase in DB04985 binding but unexpectedly not to its uptake . CONCLUSIONS : Our data support the implication of laminin receptors in cell surface binding and in transducing DB04985 anti-metastatic effects , and provide a rational for targeting cancers that express high levels of such laminin receptors . Pro-fibrogenic potential of Q9GZP0 in liver fibrosis . BACKGROUND/AIMS : We analyzed the expression of platelet-derived growth factor D ( Q9GZP0 ) in an experimental bile duct-ligated ( BDL ) rat model and assessed its biological function in cultured hepatic stellate cells ( P19526 ) and myofibroblasts ( MFB ) . METHODS : The mRNA for PDGF-A , -B , -C , -D and for PDGF receptor-alpha and -beta chains ( PDGFRalpha and PDGFRbeta ) in normal and fibrotic rat livers was assessed quantitatively . Protein levels of Q9GZP0 were quantified by immunoblotting and immunohistochemistry . RESULTS : The relative mRNA expression of all PDGF isoforms and receptors upregulated upon BDL and PDGF-A , -B and -D expression was significantly higher than that of Q9NRA1 . Q9GZP0 and PDGFRbeta protein also increased markedly . Immunostaining revealed that Q9GZP0 is localized along the fibrotic septa of the periportal- and perisinusoidal areas . Besides PDGF-B , Q9GZP0 is the second most potent PDGF isoform in PDGFRbeta signaling within P19526 /MFB , evidenced by PDGFRbeta autophosphorylation and activation of the downstream signaling molecules P27361 /2- , JNK- , p38 MAPK , and P31749 /Akt while Q9NRA1 effects were minimal . Q9GZP0 exerted mitogenic and fibrogenic effects in both cultured P19526 and MFB comparable to PDGF-B but PDGF-A and -C showed only marginal fibrogenic effects . CONCLUSIONS : Q9GZP0 possesses potential pathogenetic properties for P19526 activation and matrix remodeling in liver fibrosis . Q9GZP0 inhibition by DB05139 ameliorates tubulointerstitial fibrosis following experimental glomerulonephritis . BACKGROUND : Arresting or regressing kidney scarring is of major clinical relevance . Q9GZP0 ( Q9GZP0 ) is widely expressed in fibrotic kidneys . Administration of the Q9GZP0 neutralizing fully human monoclonal antibody DB05139 in the acute phase of progressive anti-Thy 1.1 glomerulonephritis reduced glomerular and secondary tubulointerstitial damage . METHODS : Using this model , we now assessed the effects of DB05139 ( n=15 ) vs irrelevant control IgG ( n=17 ) administered on days 17 , 28 and 35 after disease induction , i.e. after acute glomerular damage had subsided . RESULTS : In vitro , DB05139 inhibited the Q9GZP0 - but not the PDGF-B-induced proliferation of rat renal fibroblasts . Following the first DB05139 injection on day 17 , exposure to therapeutic levels was maintained until day 49 . Proteinuria in the DB05139 -treated group was transiently reduced between days 49 and 77 ( -19 to -23 % in comparison with the controls ; P < 0.05 ) . On day 100 , DB05139 treatment reduced the number of rats that had doubled their serum creatinine ( DB05139 : 40 vs controls : 71 % ; P < 0.05 ) . Compared with controls , the DB05139 animals , on day 100 , significantly lowered glomerular expression of vimentin and collagens as well as tubulointerstitial damage scores , interstitial fibrosis , vimentin and cortical Q9GZP0 mRNA levels . CONCLUSIONS : Q9GZP0 antagonism , even after the phase of acute glomerular damage , exerts beneficial effects on the course of tubulointerstitial damage , i.e. the final common pathway of most renal diseases . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . 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 . Effects of P28335 -receptor expression on cell proliferation control in hamster fibroblasts : serotonin fails to induce a transformed phenotype . 5-HT1c receptors have been shown to act as protooncogenes in NIH 3T3 cells , inducing ligand-dependent focus formation . In order to assess their mitogenic and oncogenic potential in a different cell system , we transfected these receptors into CCL39 hamster fibroblasts , a well-characterized growth factor-dependent cell line . Cell clones expressing functional receptors were isolated and tested for ( a ) growth factor dependence of proliferation measuring thymidine incorporation in response to varying doses of serum , ( b ) the response to serotonin alone or in combination with other growth factors , and ( c ) the capacity for anchorage-independent proliferation . In the absence or presence of serotonin , the large majority of the clones isolated showed normal morphology and normal growth factor dependence and was unable to grow in soft agar . None of the clones showed a significant response to serotonin alone in DNA synthesis reinitiation experiments , but synergy was observed between serotonin and the tyrosine kinase activating growth factors P01133 and FGF . However , the major part of this effect could be abolished by an antagonist of 5-HT1b receptors , which are endogenous in CCL39 cells . The same receptor was found to mediate a significant mitogenic response to the neurotransmitter in Ha-ras-transfected cells . The fact that 5-HT1c receptors do not readily induce a transformed phenotype in CCL39 cells clearly distinguishes them from strong dominantly acting oncogene products like DB01367 , P12931 , or P07333 .

MH_train_1174

MH_train_1174

MH_train_1174

interacts_with DB08827?

multiple_choice

The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . What future for combination therapies ? For most patients who require lipid-lowering treatment , statin monotherapy is the appropriate treatment . However , in those patients where statin monotherapy does not produce optimal lipid levels , the combination of a statin with niacin , a bile acid sequestrant , a fibric acid derivative , a cholesterol absorption inhibitor or a fish oil preparation may provide improved control . The choice of combination therapy depends upon the patient 's lipid profile and tolerability of the medication . Combination of a statin with niacin , a bile acid sequestrant or ezetimibe , a cholesterol absorption inhibitor , should be considered for patients with very high low-density lipoprotein cholesterol ( LDL-C ) levels , while combination with either a fibric acid derivative or a fish oil should be considered for patients with high LDL-C and high triglyceride levels . A number of new lipid-lowering agents are currently in development , including cholesteryl ester transfer protein ( P11597 ) inhibitors , acyl coenzyme A : cholesterol acyltransferase ( ACAT ) inhibitors , ileal bile acid transport ( Q12908 ) inhibitors , microsomal triglyceride transfer protein ( P55157 ) inhibitors and dual peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma agonists . Introduction of these novel therapies will provide opportunities for developing different combination strategies that may help to optimise lipid profiles in patients who are currently difficult to treat . The introduction of new combinations will require careful study to ensure that the risks of drug interactions and adverse events are minimised . 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 . 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 . Clinical experience with ramucirumab : outcomes in breast cancer . INTRODUCTION : Monoclonal antibodies and small molecules targeting the P15692 pathway are part of the arsenal to treat malignant tumors . Antiangiogenesis therapies has been studied in breast cancer with partial success , reflected by the approval of bevacizumab in Europe but not in United States , for metastatic breast cancer ( mBC ) . DB05578 is a mAb against P35968 interfering with the normal activation of this receptor by its natural ligand P15692 . AREAS COVERED : This article will review the preclinical data available to date for ramucirumab , as well as survey the main clinical trials of antiangiogenic agents reported in breast cancer , focusing on Phase III clinical trials . It will also review the clinical trial data for ramucirumab in mBC , including the design of the Phase II trials , and report on the preliminary results of the O75962 -012 trial . This trial did not meet its primary end point in progression-free survival and has to be considered as a negative trial . EXPERT OPINION : Despite preliminary positive data with ramucirumab in other metastatic solid tumors reported to date , the results of O75962 -012 discourage pursuing more efforts with ramucirumab in mBC unless predictive and reproducible biomarkers can be established to select those patients who are most likely to benefit from it . Decreasing Poly(ADP- DB01936 ) Polymerase Activity Restores ΔF508 P13569 Trafficking . Most cystic fibrosis is caused by mutations in P13569 that prevent its trafficking from the ER to the plasma membrane and is associated with exaggerated inflammation , altered metabolism , and diminished responses to oxidative stress . P09874 is activated by oxidative stress and causes energy depletion and cell dysfunction . Inhibition of this enzyme protects against excessive inflammation and recent studies have also implicated it in intracellular protein trafficking . We hypothesized that P09874 activity is altered in CF and affects trafficking and function of the most common CF mutant ΔF508 P13569 . Indeed , P09874 activity was 2.9-fold higher in CF ( ΔF508/ΔF508 ) human bronchial epithelial primary cells than in non-CF cells , and similar results were obtained by comparing CF vs. non-CF bronchial epithelial cell lines ( 2.5-fold higher in CFBE41o(-) vs. 16HBE14o(-) , P < 0.002 ) . A P09874 inhibitor ( ABT-888 , DB07232 ) partially restored P13569 channel activity in CFBE41o(-) cells overexpressing ΔF508 P13569 . Similarly , reducing P09874 activity by 85 % in ileum from transgenic CF mice ( Cftr(tm1)Eur ) partially rescued ΔF508 P13569 activity to 7 % of wild type mouse levels , and similar correction ( 7.8 % ) was observed in vivo by measuring salivary secretion . Inhibiting P09874 with ABT-888 or siRNA partially restored ΔF508 P13569 trafficking in cell lines , and most ΔF508 P13569 was complex glycosylated when heterologously expressed in P09874 (-/-) mouse embryonic fibroblasts . Finally , levels of the mature glycoform of P13569 were reduced by peroxynitrite , a strong activator of P09874 . These results demonstrate that P09874 activity is increased in CF , and identify a novel pathway that could be targeted by proteostatic correctors of P13569 trafficking . [ Use of new non-nephrotoxic immunosuppressive drugs in kidney transplantation , especially after ischemia-reperfusion injury ] . Medium- and long-term renal graft survival depends on 4 main factors : the quality of the harvested graft , ischemia-reperfusion injury during harvesting and re-implantation , rejection , and the nephrotoxicity of certain drugs ( especially immunosuppressants ) used in this setting . The most nephrotoxic immunosuppressive drugs are the anticalcineurins ( cyclosporine A and tacrolimus ) , a class discovered in the late 1970s and currently representing a basic component of all immunosuppressive protocols for solid organ graft recipients . The renal tubular and vascular toxicity of anticalcineurins is due to their immunosuppressive mechanism : they block the calcineurin pathway and thereby prevent transmission of the first signal from the T cell receptor to the nucleus , which normally triggers cytokine synthesis , New non-nephrotoxic immunosuppressants are therefore needed , especially for grafts of poor quality or subject to severe ischemia-reperfusion injury . Attention is turning to " old " molecules such as anti-thymocyte globulins , but exciting new immunosuppressants are now appearing . DB00092 is a fusion protein that binds to the immunological synapse-associated molecule P06729 , which normally interacts with LFA-3 . DB06681 , another fusion protein , blocks the T cell second signal CD 28- P33681 .1/ P33681 .2 . Finally , new chemical agents are being developed , such as sautrasporine , a tyrosine kinase inhibitor , and tofacitinib , a Jak inhibitor . Leishmania mexicana amazonensis : ADP-ribosyltransferase antagonists specifically inhibit amastigote to promastigote differentiation . Leishmania mexicana amazonensis amastigotes were induced to differentiate by incubation at 27 C . Morphological transformation was studied both in untreated cultures and in cultures where DNA synthesis , and consequently the final stage in the production of promastigotes , was inhibited by hydroxyurea . DB03073 and other antagonists of ADP-ribosyltransferase ( P09874 ) specifically inhibited differentiation at a very early stage in both experimental systems . Cell proliferation ( in the absence of hydroxyurea ) was not inhibited by P09874 antagonists -- indeed greater multiplication of undifferentiated parasites was observed in the presence of these compounds . This indicated that the parasites were being diverted from differentiation to proliferation . Preincubation of the amastigotes with the P09874 antagonists was required to produce this effect , providing further evidence that ADP-ribosylation of proteins is required for the initiation of differentiation in Leishmania . 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 . 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 . 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 . Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5-linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter ---- MLVI-2 ---- cen ---- P07686 ---- P00374 ---- Pi227- --- cp12.6 ---- ( P05113 , P05112 ) ---- P08700 ---- P04141 ---- P05230 ---- ( P07333 , P09619 ) ---- ( treC,ADRBR ) ---- ( Q5SW96 - Q13585 , P09603 ) ---- qter . The suggested order and orientation for the closely linked P08700 / P04141 gene pair is cen ---- 5' P08700 3' ---- 5' P04141 3' ---- qter , on the basis of analysis of the P04141 rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q- analyses , was telomeric to P04141 and centromeric to P07333 / P09619 , near P05230 . Long-range restriction-enzyme analysis of 5q- DNAs did not detect rearrangements of 5q-linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12.6 , P08700 , and P07333 can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions . Gateways to clinical trials . Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses . The data in the following tables has been retrieved from the Clinical Studies Knowledge Area of Prous Science Integrity(R) , the drug discovery and development portal , http://integrity.prous.com . This issue focuses on the following selection of drugs : 3,4-DAP ; DB00718 , Q16586 -10-0101 , alefacept , alemtuzumab , alosetron hydrochloride , ALT-711 , aprepitant , atazanavir sulfate , atlizumab , atvogen ; DB00188 ; P11597 vaccine , clevudine , crofelemer ; P22760 : P0C6A0 , darbepoetin alfa , decitabine , drotrecogin alfa ( activated ) , DX-9065a ; E-7010 , edodekin alfa , emivirine , emtricitabine , entecavir , erlosamide , erlotinib hydrochloride , everolimus , exenatide ; DB00569 , frovatriptan , fulvestrant ; DB00056 , gestodene ; DB04865 , human insulin ; Imatinib mesylate , indiplon , indium 111 ( 111In ) ibritumomab tiuxetan , inhaled insulin , insulin detemir , insulin glargine , ivabradine hydrochloride ; DB06792 , lapatinib , O43766 -34475 , levetiracetam , liraglutide , lumiracoxib ; Maxacalcitol , melagatran , micafungin sodium ; DB00108 , NSC-640488 ; Oblimersen sodium ; DB08439 sodium , PEG-filgrastim , peginterferon alfa-2(a) , peginterferon alfa-2b , pexelizumab , pimecrolimus , pleconaril , pramlintide acetate , pregabalin , prucalopride ; DB00025 -PFM , Ranelic acid distrontium salt , ranolazine , rDNA insulin , recombinant human soluble thrombomodulin , rhGM- P04141 , roxifiban acetate , RSD-1235 , rubitecan , ruboxistaurin mesilate hydrate ; SC-51 , squalamine ; DB01079 maleate , telbivudine , tesaglitazar , testosterone gel , tezosentan disodium , tipranavir ; DB04879 succinate ; DB04898 ; Yttrium 90 ( 90Y ) ibritumomab tiuxetan ; DB00399 monohydrate . Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV-1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE3-tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e.g. , all-trans-retinoic acid ( RA ) , 13-cis-retinoic acid ( 13- DB00982 ) , arotinoid acid ( DB02877 ) and m-carboxy-arotinoid acid ( m-carboxy- DB02877 , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 ( 24 nM ) greater than P10826 ( 4.0 nM ) greater than P13631 ( 1.3 nM ) , while the ED50 values for DB02877 and 13- DB00982 decreased in the order of P10276 ( 6.5 nM , 190 nM ) greater than P13631 ( 2.3 nM , 140 nM ) greater than P10826 ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy- DB02877 , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 greater than P10826 greater than P13631 . 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 . 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 . Lipophilic antifolate trimetrexate is a potent inhibitor of Trypanosoma cruzi : prospect for chemotherapy of Chagas ' disease . Trypanosoma cruzi , a protozoan parasite , is the causative agent for Chagas ' disease , which poses serious public health problem in Latin America . The two drugs available for the treatment of this disease are effective only against recent infections and are toxic . P00374 ( P00374 ) has a proven track record as a drug target . The lipophilic antifolate trimetrexate ( DB01157 ) , which is an FDA-approved drug for the treatment of Pneumocystis carinii infection in AIDS patients , is a potent inhibitor of T. cruzi P00374 activity , with an inhibitory constant of 6.6 nM . The compound is also highly effective in killing T. cruzi parasites . The 50 and 90 % lethal dose values against the trypomastigote are 19 and 36 nM , and the corresponding values for the amastigote form are 26 and 72 nM , respectively . However , as DB01157 is also a good inhibitor of human P00374 , further improvement of the selectivity of this drug would be preferable . Identification of a novel antifolate selective against T. cruzi would open up new therapeutic avenues for treatment of Chagas ' disease . Novel real-time polymerase chain reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia . The recent World Health Organization classification recognizes different subtypes of acute myeloid leukemia ( AML ) according to the presence of several recurrent genetic abnormalities . Detection of these abnormalities and other molecular changes is of increasing interest because it contributes to a refined diagnosis and prognostic assessment in AML and enables monitoring of minimal residual disease . These genetic abnormalities can be detected using single RT-PCR , although the screening is still labor intensive and costly . We have developed a novel real-time RT-PCR assay to simultaneously detect 15 AML-associated rearrangements that is a simple and easily applicable method for use in clinical diagnostic laboratories . This method showed 100 % specificity and sensitivity ( 95 % confidence interval , 91 % to 100 % and 92 % to 100 % , respectively ) . The procedure was validated in a series of 105 patients with AML . The method confirmed all translocations detected using standard cytogenetics and fluorescence in situ hybridization and some additional undetected rearrangements . Two patients demonstrated two molecular rearrangements simultaneously , with P11274 - P00519 implicated in both , in addition to Q01196 -MECOM in one patient and P29590 - P10276 in another . In conclusion , this novel real-time RT-PCR assay for simultaneous detection of multiple AML-associated fusion genes is a versatile and sensitive method for reliable screening of recurrent rearrangements in AML . Quality of life in plaque psoriasis patients treated with voclosporin : a Canadian phase III , randomized , multicenter , double-blind , placebo-controlled study . BACKGROUND : Quality of life assessments are important in the evaluation of new therapies for psoriasis . OBJECTIVE : To determine the effect of voclosporin ( VCS ) treatment on quality of life in patients with psoriasis . PATIENTS AND METHODS : 451 plaque psoriasis patients with ≥ 10 % body surface area involvement were randomly assigned in a double-blind fashion to 1 of 4 treatment groups ( placebo , VCS 0.2 mg kg(-1) P55957 , VCS 0.3 mg kg(-1) P55957 , and VCS 0.4 mg kg(-1) P55957 ) for up to 12 weeks of treatment . Quality of life was assessed using the Dermatology Life Quality Index ( DLQI ) and the Psoriasis Disability Index ( P07237 ) . RESULTS : At 12 weeks , patients treated with VCS 0.4 mg kg(-1) P55957 had statistically significantly more favourable assessments than placebo-treated patients in all domains of the DLQI and the P07237 . Patients treated with VCS 0.3 mg kg(-1) P55957 had statistically significant improvements in 5 of 10 domains of the DLQI and all domains of the P07237 . Patients treated with VCS 0.2 mg kg(-1) P55957 had statistically significant improvements in 4 of 10 domains of the DLQI and 2 of 4 domains of the P07237 . CONCLUSION : Treatment with VCS 0.4 mg kg(-1) P55957 significantly improves the quality of life of patients with psoriasis . Microdeletions involving chromosomes 12 and 22 associated with syndromic Duane retraction syndrome . BACKGROUND : Duane retraction syndrome ( Q9H307 ) is the most common of the congenital cranial dysinnervation disorders ( CCDDs ) . CCDDs can be monogenic or chromosomal in origin . Identification of the genetic cause(s) in patients and families with Q9H307 facilitates definitive diagnosis and provides insights into these developmental errors . MATERIALS AND METHODS : This study described a young girl with Q9H307 on the left and several additional developmental abnormalities . Clinical examination including neuroimaging , sequencing of candidate genes associated with Q9H307 , and array comparative genomic hybridization ( array CGH ) were performed . RESULTS : The proband had unilateral Q9H307 type 3 on the left with somewhat low-set ears , mild motor delay with normal intelligence , and an asymmetric neck without a palpable right sternocleidomastoid muscle . Spine X-rays revealed a Klippel-Feil syndrome ( KFS ) and an Q9BWK5 showed a webbed neck . She also had spina bifida at Q99618 -T1 and a submucosal cleft palate . The parents of the proband were related with no other family member affected similarly . Sequencing of Q9UJQ4 , P15882 , P49639 , and Q13509 did not show any mutation . Array CGH revealed de novo deletions of 21 Kb on chromosome 12q24.31 and 11 Kb on chromosome 22q13.31 , each encompassing only one gene , ring finger protein 34 , E3 ubiquitin protein ligase ( Q969K3 ) and peroxisome proliferator-activated receptor alpha ( Q07869 ) respectively . CONCLUSIONS : This patient presents an unusual phenotype associated with a unique combination of two chromosomal microdeletions . Structure and function of the Q5SW96 family of ADP-ribosyl-acceptor hydrolases . ADP-ribosylation is a post-translational protein modification , in which ADP-ribose is transferred from nicotinamide adenine dinucleotide ( NAD(+) ) to specific acceptors , thereby altering their activities . The ADP-ribose transfer reactions are divided into mono- and poly-(ADP-ribosyl)ation . Cellular ADP-ribosylation levels are tightly regulated by enzymes that transfer ADP-ribose to acceptor proteins ( e.g. , ADP-ribosyltransferases , poly-(ADP-ribose) polymerases ( PARP ) ) and those that cleave the linkage between ADP-ribose and acceptor ( e.g. , ADP-ribosyl-acceptor hydrolases ( Q5SW96 ) , poly-(ADP-ribose) glycohydrolases ( Q86W56 ) ) , thereby constituting an ADP-ribosylation cycle . This review summarizes current findings related to the Q5SW96 family of proteins . This family comprises three members ( P54922 -3 ) with similar size ( 39kDa ) and amino acid sequence . P54922 catalyzes the hydrolysis of the N-glycosidic bond of mono-(ADP-ribosyl)ated arginine . Q9NX46 hydrolyzes poly-(ADP-ribose) ( PAR ) and O-acetyl-ADP-ribose . The different substrate specificities of P54922 and Q9NX46 contribute to their unique roles in the cell . Based on a phenotype analysis of P54922 (-/-) and Q9NX46 (-/-) mice , P54922 is involved in the action by bacterial toxins as well as in tumorigenesis . Q9NX46 participates in the degradation of PAR that is synthesized by P09874 in response to oxidative stress-induced DNA damage ; this hydrolytic reaction suppresses PAR-mediated cell death , a pathway termed parthanatos . 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 . Molecular mechanisms of patupilone resistance . DB03010 is an epothilone in advanced clinical development that has shown promising efficacy in heavily pretreated patients . This study aimed at characterizing the mechanisms of patupilone activity in resistant patients . To this end , we generated patupilone-resistant cells using two cellular models , the first characterized by high chemosensitivity and low class III beta-tubulin ( Q13509 ) expression ( A2780 ) , and the second by low chemosensitivity and high Q13509 expression ( OVCAR-3 ) . The obtained cell lines were named EPO3 and OVCAR-EPO , respectively . The same selection procedure was done in A2780 cells to generate a paclitaxel-resistant cell line ( TAX50 ) . Factors of resistance are expected to increase in the drug-resistant cell lines , whereas factors of drug sensitivity will be down-regulated . Using this approach , we found up-regulation of Q13509 in TAX50 , but not EPO3 , cells , showing that Q13509 mediates the resistance to paclitaxel but not to patupilone . Moreover , Q13509 was a factor of patupilone sensitivity because OVCAR-EPO cells exhibited a dramatic reduction of Q13509 and a concomitant sensitization to hypoxia and cisplatin-based chemotherapy . To identify the mechanisms underlying patupilone resistance , tubulin genes were sequenced , thereby revealing that a prominent mechanism of drug resistance is represented by point mutations in class I beta-tubulin . Overall , these results suggest that paclitaxel and patupilone have nonoverlapping mechanisms of resistance , thus allowing the use of patupilone for those patients relapsing after paclitaxel-based chemotherapy . Furthermore , patupilone represents a promising first-line option for the treatment of high-risk ovarian cancer patients , who exhibit high Q13509 levels and poor response to standard paclitaxel-platin chemotherapy .

MH_train_1175

MH_train_1175

MH_train_1175

interacts_with DB00677?

multiple_choice

Chronic treatment with agomelatine or venlafaxine reduces depolarization-evoked glutamate release from hippocampal synaptosomes . BACKGROUND : Growing compelling evidence from clinical and preclinical studies has demonstrated the primary role of alterations of glutamatergic transmission in cortical and limbic areas in the pathophysiology of mood disorders . Chronic antidepressants have been shown to dampen endogenous glutamate release from rat hippocampal synaptic terminals and to prevent the marked increase of glutamate overflow induced by acute behavioral stress in frontal/prefrontal cortex . DB06594 , a new antidepressant endowed with MT1/ P02795 agonist and P28335 serotonergic antagonist properties , has shown efficacy at both preclinical and clinical levels . RESULTS : Chronic treatment with agomelatine , or with the reference drug venlafaxine , induced a marked decrease of depolarization-evoked endogenous glutamate release from purified hippocampal synaptic terminals in superfusion . No changes were observed in GABA release . This effect was accompanied by reduced accumulation of SNARE protein complexes , the key molecular effector of vesicle docking , priming and fusion at presynaptic membranes . CONCLUSIONS : Our data suggest that the novel antidepressant agomelatine share with other classes of antidepressants the ability to modulate glutamatergic transmission in hippocampus . Its action seems to be mediated by molecular mechanisms located on the presynaptic membrane and related with the size of the vesicle pool ready for release . P25021 mediated relaxation of buffalo ( Bubalus bubalus ) ureter . On the buffalo ureter , histamine did not elicit any direct effect . However , it caused concentration-dependent relaxation of the tissues precontracted by carbachol , phenylephrine , norepinephrine , KCI or BaCl2 and also inhibited the contractile effect of carbachol . DB08805 selectively antagonised the relaxation and inhibition of contractile response but mepyramine did not show this effect . Isoprenaline , dobutamine , salbutamol , verapamil and papaverine neither produced any direct effect nor relaxed the carbachol-contracted tissues ; norepinephrine and epinephrine had contractile effects . Hence , the histamine-induced relaxation was mediated through the activation of H2 receptors and not through adrenergic mechanisms or blockade of Ca(2+)-channels or inhibition of cyclic nucleotide phosphodiesterase . Genetic characterisation of circulating fetal cells allows non-invasive prenatal diagnosis of cystic fibrosis . OBJECTIVES : Cystic fibrosis ( CF ) is an autosomal recessive disease due to mutations in the cystic fibrosis transmembrane conductance regulator ( P13569 ) gene . The purpose of this study was to develop a molecular method to characterise both paternal and maternal P13569 alleles in DNA from circulating fetal cells ( CFCs ) isolated by ISET ( isolation by size of epithelial tumour/trophoblastic cells ) . METHODS : The molecular protocol was defined by developing the F508del mutation analysis and addressing it both to single trophoblastic cells , isolated by ISET and identified by short tandem repeats ( STR ) genotyping , and to pooled trophoblastic genomes , thus avoiding the risk of allele drop out ( Q96SZ5 ) . This protocol was validated in 100 leucocytes from F508del carriers and subsequently blindly applied to the blood ( 5 mL ) of 12 pregnant women , at 11 to 13 weeks of gestation , whose offspring had a 1/4 risk of CF . Ten couples were carriers of F508del mutation , while two were carriers of unknown P13569 mutations . RESULTS : Results showed that one fetus was affected , seven were heterozygous carriers of a P13569 mutation , and four were healthy homozygotes . These findings were consistent with those obtained by chorionic villus sampling ( CVS ) . CONCLUSION : Our data show that the ISET-CF approach affords reliable prenatal diagnosis ( P01160 ) of cystic fibrosis and is potentially applicable to pregnant women at risk of having an affected child , thus avoiding the risk of iatrogenic miscarriage . 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 . 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 . P16473 antibodies : advances and importance of detection techniques in thyroid diseases . The study of autoimmune thyroid disorders ( AITD ) has greatly contributed to our knowledge of autoimmunity . Graves ' disease and Hashimoto 's thyroiditis represent two ends of the range of autoimmune responses seen in AITD . Autoantibodies reactive to cytoplasmic antigens are associated with cell damage , and thyrotropin ( DB00024 ) -receptor antibodies ( TRAb ) influence the function and growth of the gland and play a major role in pathogenesis . The heterogeneous nature of TRAb is well accepted . Besides their long-known thyroid stimulating activity , TRAb can act as blocking antibodies or growth-promoting antibodies and , thus , cause hypothyroidism ( primary myxedema ) or endemic and sporadic goiters , respectively . Advanced methodologies for detection of these antibodies with the DB00024 -receptor assay and thyroid cell bioassay allow various activities to be measured . Current data using these assays confirm the presence of heterogeneity of functional activities of TRAb(s) in vivo . The activity of predominating antibody may relate to clinical presentation . This indicates a need for paired determinations of both DB00024 -binding inhibitory immunoglobulin ( TBII ) and thyroid-stimulating immunoglobulin ( TSI ) for accurate clinical correlations . Cloning the DB00024 -receptor gene has clarified its structure and function . The future identification of its epitopes will further delineate the clinical role of these antibodies and may allow development of new diagnostic and therapeutic approaches . Transmitter neurochemistry of the efferent neuron system innervating the labyrinth . It is likely that several mechanisms contribute to the efferent control of cochlear and vestibular function . Different effects are probably mediated by different neuronal transmitters . In spite of a number of transmitter candidates , it is still widely assumed that the entire efferent system can be globally characterized as cholinergic . We attempted to label retrogradely identified efferent neurons in the brainstem with a monoclonal antibody against choline acetyltransferase ( P28329 ) , the acetylcholine ( ACh ) synthesizing enzyme . Only a portion of the vestibular efferents could thus be shown to be cholinergic in the rat . Medial cochlear efferents , terminating under outer hair cells , may also be cholinergic since they stain intensely for acetylcholine esterase ( P22303 ) after pre-treatment with the P22303 inhibitor diisopropylfluorophosphate ( DB00677 ) . The lateral cochlear efferents terminating under inner hair cells , as well as more than half of the vestibular efferent neuron population , reacted negatively with either method designed to identify cholinergic neurons . Half of the lateral olivo-cochlear neuron population filled retrogradely with tritiated gamma-amino butyric acid [ ( 3H ] -GABA ) . These cells were similar in size and distribution to neurons staining for the GABA synthesizing enzyme glutamic acid decarboxylase ( Q99259 ) . Retrograde transport of [ 3H ] -aspartate from the inner ear to the brainstem was seen in half of the lateral olivocochlear population , as well as in part of the efferent vestibular population in group E and in the caudal pontine reticular nucleus ( P16435 ) . Since various peptides have also been located in efferent neurons , this system is chemically diversified . Several distinct mechanisms of efferent control with presumably differing functions must , therefore , exist . Autoimmune predisposition in Down syndrome may result from a partial central tolerance failure due to insufficient intrathymic expression of O43918 and peripheral antigens . Down syndrome ( DS ) , or trisomy of chromosome 21 , is the most common genetic disorder associated with autoimmune diseases . O43918 protein ( O43918 ) , a transcription factor located on chromosome 21 , plays a crucial role in autoimmunity by regulating promiscuous gene expression ( pGE ) . To investigate if autoimmunity in DS is promoted by the reduction of pGE owing to dysregulation of O43918 , we assessed the expression of O43918 and of several peripheral tissue-restricted Ag genes by quantitative PCR in thymus samples from 19 DS subjects and 21 euploid controls . Strikingly , despite the 21 trisomy , O43918 expression was significantly reduced by 2-fold in DS thymuses compared with controls , which was also confirmed by fluorescent microscopy . Allele-specific quantification of intrathymic O43918 showed that despite its lower expression , the three copies are expressed . More importantly , decreased expression of O43918 was accompanied by a reduction of pGE because expression of tissue-restricted Ags , P02708 , Q99259 , P60201 , KLK3 , SAG , TG , and P16473 , was reduced . Of interest , thyroid dysfunction ( 10 cases of hypothyroidism and 1 of Graves disease ) developed in 11 of 19 ( 57.9 % ) of the DS individuals and in none of the 21 controls . The thymuses of these DS individuals contained significantly lower levels of O43918 and thyroglobulin , to which tolerance is typically lost in autoimmune thyroiditis leading to hypothyroidism . Our findings provide strong evidence for the fundamental role of O43918 and pGE , namely , central tolerance , in the predisposition to autoimmunity of DS individuals . 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 . The response of striatal neuropeptide Y and cholinergic neurons to excitatory amino acid agonists . The effect of excitatory amino acid antagonists on antagonist on neuropeptide Y ( P01303 ) and cholinergic neurons in the striatum of the rat was studied by means of P01303 immunocytochemistry , DB00677 histochemistry for acetylcholinesterase ( P22303 ) , and biochemical determinations of choline acetyltransferase ( P28329 ) . Intrastriatal infusion of drugs revealed that striatal neurons containing P01303 are more sensitive than cholinergic neurons to the neurotoxic actions of kainic acid ( KA ) , quinolinic acid ( QA ) and L-glutamic acid ( GA ) ; all 3 compounds produced a marked loss of P01303 neurons , but only a moderate decrease in the number of P22303 neurons or P28329 activity . Co-injection experiments showed that the neurotoxicity of QA and GA , but not that of KA , can be antagonized by the specific N-methyl-D-aspartate ( DB01221 ) receptor antagonist 3-((+/-)-2-(carboxypiperazine-4-yl))-propyl-1-phosphonic acid ( CPP ) . Destruction of the glutamatergic corticostriatal projection by cerebral decortication protected striatal P01303 and cholinergic neurons against KA neurotoxicity . These results indicate that striatal P01303 and cholinergic neurons receive prominent cortical amino acid afferents , and that the neurotoxic effect of QA and GA on these neurons is mediated through DB01221 receptors . Recent insights into the molecular genetics of the homocysteine metabolism . The homocysteine plasma level is determined by non-genetic and genetic factors . In recent years evidence has accumulated that the total homocysteine plasma level of patients under different forms of renal replacement therapy is influenced by a common mutation at nucleotide position 677 of the gene coding for 5,10-methylenetetrahydrofolate reductase ( P42898 677C --> T ) . Furthermore , compound heterozygosity for the 677T allele and a novel A --> C polymorphism at nucleotide position 1298 of P42898 is suggested to correlate with a decrease of folate plasma concentrations . Because polymorphisms of genes coding for proteins involved in the metabolism of homocysteine may contribute to elevated total homocysteine plasma concentrations , molecular genetic analyses of the homocysteine pathways experienced a drift towards screening for candidate genes with a putative relationship to total homocysteine plasma levels . One example is the cloning of the P15328 gene coding for the folate-binding protein ( Folbp1 ) , which has recently been inactivated in mice , thus representing an elegant model to investigate the consequence on the homocysteine metabolism . Furthermore , the recent characterization of the O60494 gene encoding the intrinsic factor-vitamin B12 receptor ( cubilin ) provides a basis to identify the causative mutations in patients suffering from a hereditary syndrome of hyperhomocysteinemia that presents with megaloblastic anemia and proteinuria . This review focuses on recent insights into the molecular genetics of P42898 , P15328 , and O60494 , and their relationships to the metabolism of the amino acid homocysteine . Auditory function in adrenomyeloneuropathy . Auditory brainstem responses ( Q12979 ) , ipsilateral and contralateral acoustic reflexes and the masking level difference for speech ( O15121 ) were studied in 29 patients with adrenomyeloneuropathy ( Q9BXJ7 ) . Abnormalities were seen for all Q12979 components with Waves V and III affected to the greatest degree . For male patients with Q9BXJ7 , the I-III , III-V and I-V interpeak latency intervals were abnormal for a majority of patients . For female patients with Q9BXJ7 , the I-V and III-V interpeak latency intervals were abnormal for a majority of patients with the I-III interval less affected . Contralateral acoustic reflexes were elevated or absent for approximately 50 % of ears . Ipsilateral acoustic reflexes were abnormal for 25 % of ears . MLDs were significantly reduced in 72 % of patients . When considered in terms of the earliest Q12979 wave abnormality , the earlier components of the Q12979 ( i.e. , Waves III and I ) were the initial components impaired for the majority of ears . Word recognition in quiet was relatively unimpaired for all subjects . Despite the presence of marked Q12979 abnormalities , patients with Q9BXJ7 denied the presence of significant difficulty hearing . A frameshift mutation in the cubilin gene ( O60494 ) in Beagles with Imerslund-Gräsbeck syndrome ( selective cobalamin malabsorption ) . Mammals are unable to synthesize cobalamin or vitamin B12 and rely on the uptake of dietary cobalamin . The cubam receptor expressed on the intestinal endothelium is required for the uptake of cobalamin from the gut . Cubam is composed of two protein subunits , amnionless and cubilin , which are encoded by the Q9BXJ7 and O60494 genes respectively . Loss-of-function mutations in either the Q9BXJ7 or the O60494 gene lead to hereditary selective cobalamin malabsorption or Imerslund-Gräsbeck syndrome ( IGS ) . We investigated Beagles with IGS and resequenced the whole genome of one affected Beagle at 15× coverage . The analysis of the Q9BXJ7 and O60494 candidate genes revealed a homozygous deletion of a single cytosine in exon 8 of the O60494 gene ( c.786delC ) . This deletion leads to a frameshift and early premature stop codon ( p.Asp262Glufs*47 ) and is , thus , predicted to represent a complete loss-of-function allele . We tested three IGS-affected and 89 control Beagles and found perfect association between the IGS phenotype and the O60494 :c.786delC variant . Given the known role of cubilin in cobalamin transport , which has been firmly established in humans and dogs , our data strongly suggest that the O60494 :c.786delC variant is causing IGS in the investigated Beagles . DB05595 , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC(50) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 did not block membrane binding of radiolabeled folic acid , 5-methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 had a minimal effect on the cytoplasmic accumulation of 5-methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC(50) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated . Proliferative effect of histamine on MA-10 Leydig tumor cells mediated through P25021 activation , transient elevation in DB02527 production , and increased extracellular signal-regulated kinase phosphorylation levels . Mast cells ( MC ) occur normally in the testis with a species-specific distribution , yet their precise role remains unclear . Testicular MC express histidine decarboxylase ( HDC ) , the unique enzyme responsible for histamine ( HA ) generation . Evidence to date supports a role for HA as a local regulator of steroidogenesis via functional H₁ and H₂ receptor subtypes ( P35367 and P25021 , respectively ) present in Leydig cells . Given that HA is a well-known modulator of physiological and pathological proliferation in many different cell types , we aimed in the present study to evaluate whether HA might contribute to the regulation of Leydig cell number as well as to the control of androgen production . Herein , we demonstrate , to our knowledge for the first time , that MA-10 Leydig tumor cells , but not normal immature Leydig cells ( Q9Y4X3 ) , exhibit a proliferative response upon stimulation with HA that involves P25021 activation , transient elevation of DB02527 levels , and increased extracellular signal-regulated kinase ( P29323 ) phosphorylation . Our results also reveal that MA-10 cells show significantly heightened HDC expression compared to normal Q9Y4X3 or whole-testicular lysate and that inhibition of HDC activity decreases MA-10 cell proliferation , suggesting a possible correlation between autocrine overproduction of HA and abnormally increased proliferation in Leydig cells . The facts that germ cells are also both source and target of HA and that multiple testicular cells are susceptible to HA action underline the importance of the present study , which we hope will serve as a first step for further research into regulation of non-MC-related HDC expression within the testis and its significance for testicular function . Activation of chloride secretion by isoflavone genistein in endometrial epithelial cells . BACKGROUND/AIM : DB01645 , the most active isoflavone found primarily in soybeans , alters ion transport functions in intestinal and airway epithelia . The present study aims to investigate the acute effects and mechanisms of action of genistein in immortalized porcine endometrial epithelial cells . METHODS : Ussing chamber technique was used for transepithelial electrical measurements . RESULTS : DB01645 increased short-circuit currents ( Isc ) which were inhibited by glibenclamide , P16860 , CFTRinh-172 , DIDS or bumetanide , but not amiloride . In experiments with amphotericin B-permeabilized monolayers , genistein activated the apical Cl- current and barium-sensitive basolateral K+ current while inhibiting the apical K+ current . DB01645 failed to increase the Isc in the presence of forskolin or DB07954 , but did increase the Isc in UTP . Pretreatment with genistein also abolished the increase in the Isc when induced by forskolin , DB07954 or UTP . However , Ca2+-chelating BAPTA-AM did not affect the genistein-induced increase in the Isc . The genistein-stimulated Isc was reduced by tyrosine kinase inhibitors , tyrphostin A23 or AG490 . However , vanadate , a tyrosine phosphatase inhibitor , failed to inhibit the genistein response . P03372 antagonist ICI182,780 did not alter the genistein’s action . CONCLUSION : The soy isoflavone , genistein , stimulates Cl- secretion in endometrial epithelial cells possibly via a direct activation of P13569 which appears to be modulated through a tyrosine kinase-dependent pathway . The present findings may be of benefit for the therapeutic application of genistein in the treatment of electrolyte transport disorders in the epithelia . DB02533 reduces cisplatin ototoxicity . DB00515 is known to cause high-frequency neurosensory hearing loss . While reactive oxygen species have been shown to play a role , reactive nitrogen species have been implicated , but not proven to be involved , in cisplatin ototoxicity . The purpose of the present study was to investigate the role of nitric oxide ( *NO ) in cisplatin ototoxicity by administering aminoguanidine ( AG ) , a relatively specific inhibitor of inducible nitric oxide synthase ( P35228 ) , in conjunction with cisplatin . Rats were injected with cisplatin , AG , or both . Auditory brainstem evoked responses ( Q12979 ) were measured before and 3 days after cisplatin administration . The cochlear tissue was then assayed for *NO and malondialdehyde . DB00515 alone caused significant Q12979 threshold shifts at all stimuli tested , whereas AG alone caused no shifts . There was a significant reduction in threshold shift for clicks and 16 kHz tone bursts ( but not 32 kHz ) when AG was given with cisplatin . The malondialdehyde concentration ( but not the *NO concentration ) in the AG/cisplatin group was significantly lower than that of the cisplatin group . This suggests that AG reduces cisplatin ototoxicity by directly scavenging hydroxyl radicals . The P35228 pathway may play a role in the generation of free radicals and hearing loss resulting from cisplatin administration , but this conclusion was not supported by our data . A novel extract SB-300 from the stem bark latex of Croton lechleri inhibits P13569 -mediated chloride secretion in human colonic epithelial cells . An oligomeric proanthocyanidin ( DB04941 ) extracted from the bark latex of the tree Croton lechleri ( family Euphorbiaceae ) is a potent inhibitor of cholera toxin-induced fluid accumulation and chloride secretion . The manufacturing process for DB04941 was optimized and simplified to produce an increased yield of the herbal extract . The novel extract ( named SB-300 ) contained on average 70.6+/-7.2 % DB04941 by weight ( mean +/- S.D. ; n=56 lots ) . Here , we describe the effectiveness of SB-300 on DB02527 -regulated chloride secretion , which is mediated by the cystic fibrosis transmembrane conductance regulator Cl- channel ( P13569 ) in human colonic T84 cells . Exposure of the apical surface to SB-300 blocked forskolin-stimulated Cl- secretion by 92.2+/-3.0 % with a half-maximal inhibition constant ( KB ) of 4.8+/-0.8 microM . For DB04941 , stimulated Cl- currents were decreased by 98.0+/-7.2 % and KB averaged 4.1+/-1.3 microM . There was no significant difference between the blocking kinetics of DB04941 and SB-300 . DB02587 -stimulated whole cell Cl- currents were effectively blocked by extracellular addition of SB-300 ( 63+/-8.5 % ; n=3 ) and to a similar extent by DB04941 ( 83 +/- 0.6 % ; n=2 ; at 50 microM each ) . Both extracts inhibited a time- and voltage-independent Cl- conductance , which indicated the involvement of P13569 Cl- channels . We conclude that both DB04941 ( used in DB04941 ) and SB-300 ( used in P46459 Normal Stool Formula ) are novel natural products that target the P13569 Cl- channel . SB-300 is a low cost herbal extract and may present a complementary and alternative medicine approach for the treatment of fluid loss in watery diarrhea . 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 . Alteration of N-ras gene mutation after relapse in acute lymphoblastic leukemia . We investigated N-ras activation in childhood acute lymphoblastic leukemia ( dALL ) by the polymerase chain reaction ( PCR ) and the oligonucleotide hybridization method . The frequency of point-mutation of the N-ras gene was not high ( 2 of 15 ) , and one positive case who relapsed was analyzed in detail . Although N-ras gene activation was detected at both onset and relapse , the mutation sites were different . At onset , DB00145 ( P19440 ) was changed to DB00133 ( AGT ) at codon 12 , and at relapse , DB00145 ( P19440 ) to DB00128 ( Q6IB77 ) was observed at the same codon . In addition , the DNA at relapse showed a remarkably higher transforming activity than the DNA at onset on two independent recipient cell lines . The identical cell surface phenotype and the same rearrangement patterns of both the immunoglobulin ( Ig ) heavy chain and T-cell receptor ( TCR ) gamma chain genes indicated that the leukemic cells at onset and those at relapse were derived from the same precursor cell . Therefore , this case supports the concept that ras activation is not the event initiating leukemogenesis , but may be involved in leukemic progression . Changes in urinary proximal tubule parameters in neonatal rats exposed to cadmium chloride during pregnancy . Pregnant Sprague-Dawley rats were injected intraperitoneally with physiological saline solution ( vehicle ) or cadmium chloride ( CdCl2 ) at 2.0 or 2.5 mg kg-1 on days 8 , 10 , 12 and 14 of gestation . On postnatal day ( P01160 ) 3 , 12 or 49 , the offspring were examined for 8- or 24-h urinary excretion of beta 2-microglobulin ( beta 2-m ) , metallothionein ( MT ) and urinary activity of three proximal tubular enzymes : gammaglutamyl transferase ( P19440 ) , alkaline phosphatase ( ALP ) and O60502 ( NAG ) . Treatment with CdCl2 did not affect growth or survival of offspring . Significant decreases in the urinary excretion of P19440 , ALP and NAG were observed on P01160 3 , at both doses . Exposure to 4 x 2.5 mg kg-1 resulted in functional deficit of the proximal tubule on P01160 3 , as evidenced by the significant increase in beta 2-m . Except for a slight but significant increase of beta 2-m in 49-day-old males , all the other urinary parameters returned to control values on P01160 12 . There was no effect on MT . Results from this study show that prenatal exposure to CdCl2 can induce significant changes in the kidney biochemistry of rats in the early postnatal period . 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 . The dual role of serotonin in defense and the mode of action of antidepressants on generalized anxiety and panic disorders . Antidepressants are widely used to treat several anxiety disorders , among which generalized anxiety disorder ( Q99259 ) and panic disorder ( PD ) . Serotonin ( 5-HT ) is believed to play a key role in the mode of action of these agents , a major question being which pathways and receptor subtypes are involved in each type of anxiety disorder . The dual role of 5-HT in defense hypothesis assumes that 5-HT facilitates defensive responses to potential threat , like inhibitory avoidance , related to anxiety , whereas it inhibits defensive responses to proximal danger , like one-way escape , related to panic . The former action would be exerted at the forebrain , chiefly the amygdala and medial prefrontal cortex ( P27918 ) , while the latter would be exerted at the dorsal periaqueductal gray ( DPAG ) matter of the midbrain . The present review is focused on studies designed to test this hypothesis , performed in animal models of anxiety and panic , as well as in human experimental anxiety tests . The reviewed results suggest that chronic , but not acute , administration of antidepressants suppress panic attacks by increasing the release of 5-HT and enhancing the responsivity of post-synaptic P08908 and 5- Q13049 receptors in the DPAG . The attenuation of generalized anxiety , also caused by the same drug treatment , would be due to the desensitization of P28335 receptors and , less certainly , to increased stimulation of P08908 receptors in forebrain structures . This action would result in less activation of the amygdala , medial P27918 and insula by warning signals , as shown by the reviewed results obtained with functional neuroimaging in healthy volunteers and patients with anxiety disorders .

MH_train_1176

MH_train_1176

MH_train_1176

interacts_with DB01200?

multiple_choice

Analysis of CAD gene amplification using a combined approach of molecular genetics and cytogenetics . CAD is a multifunctional protein which catalyzes the first three steps of de novo uridine biosynthesis . Rodent cells resistant to DB03459 , a specific inhibitor of the ATCase activity of CAD , overproduce the P27708 and CAD mRNA as a direct result of the amplification of the CAD gene . In order to study the mechanism of CAD gene amplification , a functional Syrian hamster CAD gene was inserted into a cosmid vector using molecular cloning techniques . The cloned genes were assayed for biological function by fusing CAD-deficient Chinese hamster ovary ( CHO ) cell mutants with protoplasts of E. coli containing the CAD cosmids . Two clones with functional CAD genes were isolated and shown to contain inserts 40 and 45 kb long . The cloned genes could also be introduced into wild type CHO cells by selecting for cells which became resistant to high DB03459 concentrations in a single step . Transformations of mutant and wild type CHO cells contained multiple active copies of the donated Syrian hamster CAD genes in addition to their endogenous CHO CAD genes . The cloned genes in all transformants analyzed are integrated into host cell chromosomes at single locations defined by in situ hybridization . Independently isolated transformants contain the donated genes in different chromosomes . Co-transformation of CHO cells with two different genes by protoplast fusion is also shown to be possible . 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 ) Rheb protein binds CAD ( carbamoyl-phosphate synthetase 2 , aspartate transcarbamoylase , and dihydroorotase ) protein in a GTP- and effector domain-dependent manner and influences its cellular localization and carbamoyl-phosphate synthetase ( CPSase ) activity . Rheb small GTPases , which consist of Rheb1 and Q8TAI7 ( also known as RhebL1 ) in mammalian cells , are unique members of the Ras superfamily and play central roles in regulating protein synthesis and cell growth by activating P42345 . To gain further insight into the function of Rheb , we carried out a search for Rheb-binding proteins and found that Rheb binds to P27708 ( carbamoyl-phosphate synthetase 2 , aspartate transcarbamoylase , and dihydroorotase ) , a multifunctional enzyme required for the de novo synthesis of pyrimidine nucleotides . CAD binding is more pronounced with Q8TAI7 than with Rheb1 . Rheb binds CAD in a GTP- and effector domain-dependent manner . The region of CAD where Rheb binds is located at the C-terminal region of the carbamoyl-phosphate synthetase domain and not in the dihydroorotase and aspartate transcarbamoylase domains . Rheb stimulated carbamoyl-phosphate synthetase activity of CAD in vitro . In addition , an elevated level of intracellular UTP pyrimidine nucleotide was observed in Tsc2-deficient cells , which was attenuated by knocking down of Rheb . Immunostaining analysis showed that expression of Rheb leads to increased accumulation of CAD on lysosomes . Both a farnesyltransferase inhibitor that blocks membrane association of Rheb and knockdown of Rheb mislocalized CAD . These results establish CAD as a downstream effector of Rheb and suggest a possible role of Rheb in regulating de novo pyrimidine nucleotide synthesis . Serotonin promotes tumor growth in human hepatocellular cancer . In addition to its function as a neurotransmitter and vascular active molecule , serotonin is also a mitogen for hepatocytes and promotes liver regeneration . A possible role in hepatocellular cancer has not yet been investigated . Human hepatocellular cancer cell lines Huh7 and HepG2 were used to assess the function of serotonin in these cell lines . Characteristics of autophagy were detected with transmission electron microscopy , immunoblots of microtubule-associated protein light chain 3(LC3) and p62 ( sequestosome 1 ) . Immunoblots of the mammalian target of rapamycin ( P42345 ) and its downstream targets p70S6K and Q13541 were used to investigate signaling pathways of serotonin . Two different animal models served as principle of proof of in vitro findings . Clinical relevance of the experimental findings was evaluated with a tissue microarray from 168 patients with hepatocellular carcinoma . Serotonin promotes tumor growth and survival in starved hepatocellular carcinoma cells . During starvation hepatocellular carcinoma cells exhibited characteristics of autophagy , which disappeared in serotonin-treated cells . DB00877 , an inhibitor of P42345 , is known to induce autophagy . Serotonin could override rapamycin by an P42345 -independent pathway and activate common downstream signals such as p70S6K and Q13541 . In two tumor models of the mouse , inhibition of serotonin signaling consistently impaired tumor growth . Human biopsies revealed expression of the serotonin receptor P41595 , correlating with downstream signals , e.g. , phosphorylated p70S6K and proliferation . CONCLUSION : This study provides evidence that serotonin is involved in tumor growth of hepatocellular cancer by activating downstream targets of P42345 , and therefore serotonin-related pathways might represent a new treatment strategy . 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 . Involvement of retinoic acid receptor alpha in the stimulation of tissue-type plasminogen-activator gene expression in human endothelial cells . Retinoids stimulate tissue-type plasminogen-activator ( t-PA ) gene expression in human endothelial cells , and are likely to do so by binding to one or more nuclear retinoid receptors . The present study was initiated to identify the retinoid receptor(s) involved in this process . Expression and regulation of retinoic acid receptors ( RARs ) and retinoid X receptors ( RXRs ) were analyzed by Northern-blot analysis of total or poly(A)-rich RNA prepared from cultured human umbilical vein endothelial cells ( HUVEC ) . Prior to any exposure to retinoids , HUVEC express two transcripts for P10276 ( 3.6 kb and 2.8 kb ) , and low levels of transcripts for P10826 ( 3.4 kb and 3.2 kb ) and P13631 ( 3.3 kb and 3.1 kb ) . Two RXR subtypes were identified , RXR-alpha ( 4.8 kb ) and , at a much lower concentration , RXR-beta ( 2.4 kb ) ; no evidence for the presence of RXR-gamma was found . Furthermore , HUVEC express cellular retinol-binding protein I ( P09455 ) and cellular retinoic-acid-binding protein I ( P29762 ) mRNA . Exposure of HUVEC to 1 microM retinoic acid or the DB04942 , Ch55 , led to the induction of the two P10826 mRNAs , RXR-alpha mRNA and P09455 mRNA , whereas the expression of the other receptor and P29762 transcripts did not change appreciably . Using retinoid analogues that bind preferentially to one of the RAR or RXR subtypes , we found evidence that P10276 is involved in the retinoid-induced t-PA expression in HUVEC . This conclusion was strengthened by experiments in which blocking of P10276 with a specific P10276 antagonist , Ro 41-5253 , was demonstrated to suppress the induction of t-PA by retinoids . 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 . 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 . At least seven ribosomal proteins are involved in the control of translational accuracy in a eukaryotic organism . In the filamentous fungus Podospora anserina , ribosomal proteins of 60 mutants impaired in the control of translational fidelity have been submitted to electrophoretic analysis . The " four corners " system combining four different two-dimensional polyacrylamide gel electrophoretic systems has been used . An altered electrophoretic pattern has been observed for 12 mutants . In mutants su3 , su12 and su11 ( decreased translational fidelity ) , proteins S1 , S7 and S8 , respectively , are altered . For AS mutants ( increased translational fidelity ) , proteins S9 , P28222 and S19 , respectively , are altered in AS9 , AS1 and AS6 mutants , and protein S29 is lacking in Q9NTI5 mutants . The data suggest that five of these genes ( at least ) are the structural genes for the relevant proteins ( su3:S1 , su12:S7 , AS1: P28222 , AS6:S19 , AS9:S9 ) , while the Q9NTI5 gene may code for a modifying enzyme . 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 . 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 . 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 . Novel splice variants of the bovine P35558 gene . DB01819 carboxykinase 1 ( P35558 ) , also named P35558 , is a multiple-function gene that is involved in gluconeogenesis , glyceroneogenesis , reproduction , female fertility , and development of obesity and diabetes . How its many functions are regulated was largely unknown . Therefore , we investigated mRNA expression and possible splice variants of P35558 by screening cDNA in nine tissues from Holstein bulls and cows . P35558 mRNA was highly expressed in the liver , kidney , ovary and testis ; expression levels were low in the heart , spleen , and lung tissues . Expression of this gene was not detected in skeletal muscle . This led to the discovery of five novel bovine splice variants , named P35558 -AS1- P35558 -AS5 . In P35558 -AS1 , 51 nucleotides in the interior of exon 2 were spliced out . In P35558 -AS2 , exons 2 and 3 were altered by the alternative 3' and 5' splice sites , respectively . P35558 - Q9NTI5 was truncated from the 3' end of exon 2 to the 5' end of exon 4 . In P35558 -AS4 , exon 5 was completely spliced out . In P35558 -AS5 , exons 5 and 6 and the 5' end of exon 7 were spliced out . These splice variants ( P35558 -AS1- P35558 -AS5 ) potentially encoded shorter proteins ( 605 , 546 , 373 , 246 and 274 amino acids , respectively ) , when compared to the complete protein ( 622 amino acids ) . Considering the functional domains of the P35558 protein , it is likely that these splice variants considerably affect the function of this protein ; alternative splicing could be one of the mechanisms by which the diverse functions of P35558 are regulated . Inhibition of estrogen receptor alpha expression and function in MCF-7 cells by kaempferol . DB00286 are mitogenic for estrogen receptor ( ER ) -positive breast cancer cells . Current treatment of ER-positive breast tumors is directed towards interruption of estrogen activity . We report that treatment of ER-positive breast cancer cells with kaempferol resulted in a time- and dose-dependent decrease in cell number . The concentration required to produce 50 % growth inhibition at 48 h was approximately 35.0 and 70.0 microM for ER-positive and ER-negative breast cancer cells , respectively . For MCF-7 cells , a reduction in the P03372 mRNA equivalent to 50 , 12 , 10 % of controls was observed 24 h after treatment with 17.5 , 35.0 , and 70.0 microM of kaempferol , respectively . Concomitantly , these treatments led to a 58 , 80 , and 85 % decrease in P03372 protein . The inhibitory effect of kaempferol on P03372 levels was seen as early as 6 h post-treatment . Kaempferol treatment also led in a dose-dependent decrease in the expression of progesterone receptor ( PgR ) , cyclin D1 , and insulin receptor substrate 1 ( P35568 ) . Immunocytochemical study revealed that P03372 protein in kaempferol-treated MCF-7 cells formed an aggregation in the nuclei . Kaempferol also induced degradation of P03372 by a different pathway than that were observed for the antiestrogen DB00947 and estradiol . Estradiol-induced MCF-7 cell proliferation and expression of the estrogen-responsive-element-reporter gene activity were abolished in cells co-treated with kaempferol . These findings suggest that modulation of P03372 expression and function by kaempferol may be , in part , responsible for its anti-proliferative effects seen in in vitro . Effects of an alpha 7-nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3-(2,4-dimethoxybenzylidene)-anabaseine ( DB05708 ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 , the α7-nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 genotype . CONCLUSIONS : The observed DB05708 -related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7-nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents . 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 . 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 . Fraction from wax apple [ Syzygium samarangense ( Blume ) Merrill and Perry ] fruit extract ameliorates insulin resistance via modulating insulin signaling and inflammation pathway in tumor necrosis factor α-treated FL83B mouse hepatocytes . Inflammation is associated with the development of insulin resistance in Type 2 diabetes mellitus . In the present study , mouse FL83B cells were treated with tumor necrosis factor-alpha ( P01375 -α ) to induce insulin resistance , and then co-incubated with a fraction from wax apple fruit extract ( FWFE ) . This fraction significantly increased the uptake of the nonradioactive fluorescent indicator 2- [ N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino ] -2-deoxy-d-glucose ( 2-NBDG ) in insulin resistant cells . Western blot analysis revealed that , compared with the P01375 -α-treated control group , FWFE increased the expression of the insulin receptor ( IR ) , insulin receptor substrate-1 ( P35568 ) , protein kinase B ( Akt/ P31749 ) , phosphatidylinositol-3 kinase ( PI3K ) , and glucose transporter 2 ( P11168 ) , and increased IR tyrosyl phosporylation , in insulin resistant FL83B cells . However , FWFE decreased phosphorylation of c-Jun N-terminal kinases ( JNK ) , but not the expression of the intercellular signal-regulated kinases ( P29323 ) , in the same cells . These results suggest that FWFE might alleviate insulin resistance in P01375 -α-treated FL83B cells by activating PI3K-Akt/ P31749 signaling and inhibiting inflammatory response via suppression of JNK , rather than P29323 , activation . DNA methylation profiles in diffuse large B-cell lymphoma and their relationship to gene expression status . In an initial epigenetic characterization of diffuse large B-cell lymphoma ( DLBCL ) , we evaluated the DNA methylation levels of over 500 CpG islands . Twelve CpG islands ( AR , P49918 , DLC1 , P14416 , P43694 , P39905 , Q13224 , P42898 , P15172 , Q13562 , O95948 and P05549 ) showed significant methylation in over 85 % of tumors . Interestingly , the methylation levels of a CpG island proximal to FLJ21062 differed between the activated B-cell-like ( ABC-DLBCL ) and germinal center B-cell-like ( GCB-DLBCL ) subtypes . In addition , we compared the methylation and expression status of 67 genes proximal ( within 500 bp ) to the methylation assays . We frequently observed that hypermethylated CpG islands are proximal to genes that are expressed at low or undetectable levels in tumors . However , many of these same genes were also poorly expressed in DLBCL tumors where their cognate CpG islands were hypomethylated . Nevertheless , the proportional reductions in Q12983 , P16455 , RBP1 , P43694 , Q9BY67 , P29762 and FLJ21062 expression with increasing methylation suggest that epigenetic processes strongly influence these genes . Lastly , the moderate expression of several genes proximal to hypermethylated CpG tracts suggests that DNA methylation assays are not always accurate predictors of gene silencing . Overall , further investigation of the highlighted CpG islands as potential clinical biomarkers is warranted . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Acute pharmacodynamic and antivascular effects of the vascular endothelial growth factor signaling inhibitor DB04849 in Calu-6 human lung tumor xenografts . The vascular endothelial growth factor-A ( P15692 ) signaling pathway , a key stimulant of solid tumor vascularization , is primarily dependent on the activation of the endothelial cell surface receptor P15692 receptor-2 ( P35968 ) . DB04849 is an oral , highly potent small-molecule inhibitor of VEGFR tyrosine kinase activity that inhibits angiogenesis and the growth of human tumor xenografts in vivo . Here , we show pharmacodynamic changes in P35968 phosphorylation induced by DB04849 . In mouse lung tissue , a single dose of DB04849 at 6 mg/kg inhibited P15692 -stimulated P35968 phosphorylation by 87 % at 2 h with significant inhibition ( > or=60 % ) maintained to 24 h . To examine inhibition of P35968 phosphorylation in tumor vasculature by immunohistochemistry , a comprehensive assessment of antibodies to various phosphorylation sites on the receptor was undertaken . Antibodies to the phosphotyrosine epitopes pY1175/1173 and pY1214/1212 were found suitable for this application . Calu-6 human lung tumor xenografts , from mice receiving DB04849 or vehicle treatment ( p.o. , once daily ) , were examined by immunohistochemistry . A significant reduction in tumor vessel staining of phosphorylated P35968 ( pVEGFR-2 ) was evident within 28 h of DB04849 treatment ( 6 mg/kg ) . This effect preceded a significant reduction in tumor microvessel density , which was detectable following 52 h of DB04849 treatment . These data show that DB04849 is a potent inhibitor of P35968 activation in vivo and suggest that DB04849 delivers therapeutic benefit in Calu-6 tumors by targeting vessels dependent on P35968 signaling for survival . In addition , this work highlights the utility of measuring either pY1175/1173 or pY1214/1212 on P35968 as a pharmacodynamic marker of P35968 activation . 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 . 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 .

MH_train_1177

MH_train_1177

MH_train_1177

interacts_with DB06822?

multiple_choice

The maintenance of cisplatin- and paclitaxel-induced mechanical and cold allodynia is suppressed by cannabinoid CB₂ receptor activation and independent of P61073 signaling in models of chemotherapy-induced peripheral neuropathy . BACKGROUND : Chemotherapeutic agents produce dose-limiting peripheral neuropathy through mechanisms that remain poorly understood . We previously showed that AM1710 , a cannabilactone CB₂ agonist , produces antinociception without producing central nervous system ( CNS ) -associated side effects . The present study was conducted to examine the antinociceptive effect of AM1710 in rodent models of neuropathic pain evoked by diverse chemotherapeutic agents ( cisplatin and paclitaxel ) . A secondary objective was to investigate the potential contribution of alpha-chemokine receptor ( P61073 ) signaling to both chemotherapy-induced neuropathy and CB₂ agonist efficacy . RESULTS : AM1710 ( 0.1 , 1 or 5 mg/kg i.p. ) suppressed the maintenance of mechanical and cold allodynia in the cisplatin and paclitaxel models . Anti-allodynic effects of AM1710 were blocked by the CB₂ antagonist AM630 ( 3 mg/kg i.p. ) , but not the P21554 antagonist AM251 ( 3 mg/kg i.p. ) , consistent with a CB₂-mediated effect . By contrast , blockade of P61073 signaling with its receptor antagonist DB06809 ( 10 mg/kg i.p. ) failed to attenuate mechanical or cold hypersensitivity induced by either cisplatin or paclitaxel . Moreover , blockade of P61073 signaling failed to alter the anti-allodynic effects of AM1710 in the paclitaxel model , further suggesting distinct mechanisms of action . CONCLUSIONS : Our results indicate that activation of cannabinoid CB₂ receptors by AM1710 suppresses both mechanical and cold allodynia in two distinct models of chemotherapy-induced neuropathic pain . By contrast , P61073 signaling does not contribute to the maintenance of chemotherapy-induced established neuropathy or efficacy of AM1710 . Our studies suggest that CB₂ receptors represent a promising therapeutic target for the treatment of toxic neuropathies produced by cisplatin and paclitaxel chemotherapeutic agents . 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 . 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 . Suppression of 3-deoxyglucosone and heparin-binding epidermal growth factor-like growth factor mRNA expression by an aldose reductase inhibitor in rat vascular smooth muscle cells . Reactive carbonyl compounds and oxidative stress have been recently shown to up-regulate the expression of heparin-binding epidermal growth factor-like growth factor ( HB- P01133 ) , a potent mitogen for vascular smooth muscle cells ( SMCs ) produced by SMC themselves . Because the polyol pathway has been reported to influence the formation of carbonyl compounds and the oxidative stress in various cells , we conducted this study to investigate whether the polyol pathway affects HB- P01133 expression along with the generation of carbonyl compounds and the oxidative stress in SMCs . We found that , compared with those cultured with 5.5mM glucose , SMCs cultured with 40 mM glucose showed the accelerated thymidine incorporation , elevated levels of intracellular sorbitol , 3-deoxyglucosone ( 3-DG ) , advanced glycation end products ( AGEs ) , and thiobarbituric acid-reactive substances ( TBARS ) along with the enhanced expression of HB- P01133 mRNA . An aldose reductase inhibitor ( Q9Y4X5 ) , Q9NYY3 -860 , significantly inhibited all of these abnormalities , while aminoguanidine suppressed 3-DG levels and HB- P01133 mRNA expression independent of sorbitol levels . The results suggest that the polyol pathway may play a substantial role in SMC hyperplasia under hyperglycemic condition in part by affecting HB- P01133 mRNA expression via the production of carbonyl compounds and oxidative stress . Role of nucleotide sequences in the V3 region in efficient replication of P51681 -utilizing human immunodeficiency virus type 1 in macrophages . Macrophages express both P61073 and P51681 coreceptors , but restrict X4 HIV-1 replication unless the Env-V3 region , a major determinant of cell tropism , is exchanged with that of R5 HIV-1 . As the V3 exchange concomitantly alters the nucleotide sequences , we introduced silent mutations in the V3 or P06681 region of macrophage-tropic R5 JRFL without changing the amino acids . Immunoblot analysis confirmed that viral proteins including Env-gp120 were similarly incorporated in wild-type ( wt ) and mutant virions . The silent mutants infected P51681 -positive MAGIC5 cells but not P51681 -negative MAGI cells , as productively as wt viruses , indicating that the silent mutations did not alter coreceptor utilization . In contrast , two of three silent V3-mutant viruses failed to replicate efficiently in primary macrophages , whereas other V3- or P06681 -mutants and wt JRFL infected macrophages productively . Furthermore , synthesis of the full-length viral DNA of the aberrant V3-mutant was largely reduced in macrophages . These results suggest that V3 nucleotide sequences may be one of the postentry factors restricting HIV-1 replication in macrophages . beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells . Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention , the molecular mechanisms underlying such chemopreventive activity are not clear . Since peroxisome proliferator-activated receptor gamma ( P37231 ) has been implicated in regulating breast cancer cell differentiation and apoptosis , the effects of beta-carotene on the P37231 -mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study . The results demonstrated that beta-carotene significantly increased P37231 mRNA and protein levels in time-dependent manner . In addition , beta-carotene increased the cyclin-dependent kinase inhibitor P38936 ( P38936 /CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression . DB07863 ( GW9662 ) , an irreversible P37231 antagonist , partly attenuated the cell death caused by beta-carotene . Further , reactive oxygen species ( ROS ) production was induced by beta-carotene , resulting in mitochondrial dysfunction and cytochrome C release . DB00143 ( DB00143 ) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene . In total , these observations suggest that the synergistic effect of P37231 expression and ROS production may account for beta-carotene-mediated anticancer activities . Dissociation between stem cell phenotype and NOD/SCID repopulating activity in human peripheral blood P28906 (+) cells after ex vivo expansion . OBJECTIVE : The relationship between phenotype and function in ex vivo-cultured human hematopoietic stem cells ( P19526 ) remains poorly understood . We investigated the effects of a short-term serum-free culture on the relationship between stem cell phenotype , cell division history , and function in human P28906 (+) cells . METHODS : G- P04141 -mobilized peripheral P28906 (+) cells were cultured for 4 days with stem cell factor , flt-3 ligand , and thrombopoietin . The phenotype ( P28906 , P28907 , HLA-DR , c-kit ) , cell division history , colony-forming cell ( Q15814 ) , long-term culture-initiating cell ( LTC-IC ) , and NOD/SCID repopulating activities were evaluated at Day 0 and 4 . RESULTS : We observed a loss of P28907 , HLA-DR , and c-kit surface expression resulting in a drastic increase in P28906 (+) P28907 (-) , P28906 (+)HLA-DR(-) , and P28906 (+)c-kit(-/low) cells at Day 4 . In contrast , the frequency of Thy-1(+) cells was maintained . We observed a 1.3-fold expansion of Q15814 , a 4.8-fold increase in LTC-IC , and an overall maintenance of the NOD/SCID repopulating cell activity . P28906 (+) P28907 (-) and P28906 (+)HLA-DR(-) cells detected at Day 4 displayed the most active pattern of division ( 4 to 5 divisions ) whereas 60 % of P28906 (+)Thy-1(+) cells divided 0 to 2 times during the same period . At Day 4 , the NOD/SCID repopulating activity was associated with Thy-1(+) cells with no more than 2 divisions . CONCLUSIONS : Our results show that the relationship between stem cell phenotype and function is dramatically altered in cultured P28906 (+) cells . Thy-1 expression and cell division history appear to be superior to P28907 , HLA-DR , and c-kit , or to homing molecules ( P61073 , VLA-4 ) as predictors of the repopulating activity of cultured peripheral P28906 (+) cells . 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 . The relationship of in vivo central P21554 receptor occupancy to changes in cortical monoamine release and feeding elicited by P21554 receptor antagonists in rats . RATIONALE : Cannabinoid type 1 ( CB(1) ) receptor antagonists are reportedly effective in reducing food intake both preclinically and clinically . This may be due in part to their effects on monoamine release in the brain . The level of central CB(1) receptor occupancy underlying these neurobiological effects is unclear . OBJECTIVES : We explored the relationship between in vivo CB(1) receptor occupancy in the frontal cortex and changes in both monoamine release in the medial prefrontal cortex ( mPFC ) and feeding behavior in rats in response to two orally administered CB(1) receptor antagonists presently in clinical trials , SR141716A ( rimonabant ) and DB05077 . METHODS : CB(1) receptor occupancy was measured using [ (3)H ] SR141716A , and these occupancies were related to potencies to mediate increases in dopamine ( DA ) and norepinephrine ( NE ) release measured with microdialysis and decreases in consumption of a highly palatable diet ( HP ) . RESULTS : High receptor occupancy levels ( > 65 % ) were required to detect increases in monoamine release that were achieved with 3 and 10 mg/kg of SR141716A and 10 mg/kg of DB05077 for DA and 10 mg/kg of SR141716A for NE . Decreases in HP consumption were seen at occupancies higher than 65 % for SR141716A that were achieved with 3 and 10 mg/kg . In contrast , decreases in HP consumption were seen at relatively low CB(1) receptor occupancies ( 11 % ) for DB05077 . CONCLUSIONS : The occupancy method described here is an effective tool for interrelating central CB(1) receptor occupancy with neurobiological actions of CB(1) receptor antagonists and for furthering our understanding of the role of CB(1) receptors in central nervous system physiology and pathology . Mechanism of interaction of niflumic acid with heterologously expressed kidney CLC-K chloride channels . CLC-K Cl(-) channels belong to the CLC protein family . In kidney and inner ear , they are involved in transepithelial salt transport . Mutations in ClC-Kb lead to Bartter 's syndrome , and mutations in the associated subunit barttin produce Bartter 's syndrome and deafness . We have previously found that 3-phenyl-CPP blocks hClC-Ka and rClC- P04264 from the extracellular side in the pore entrance . Recently , we have shown that niflumic acid ( DB04552 ) , a nonsteroidal anti-inflammatory fenamate , produces biphasic behavior on human CLC-K channels that suggests the presence of two functionally different binding sites : an activating site and a blocking site . Here , we investigate in more detail the interaction of DB04552 on CLC-K channels . Mutants that altered block by 3-phenyl-2-(p-chlorophenoxy)propionic acid ( CPP ) had no effect on DB04552 block , indicating that the inhibition binding site of DB04552 is different from that of 3-phenyl-CPP and flufenamic acid . Moreover , DB04552 does not compete with extracellular Cl(-) ions , suggesting that the binding sites of DB04552 are not located deep in the pore . Differently from ClC-Ka , on the rat homologue P51800 , DB04552 has only an inhibitory effect . We developed a quantitative model to describe the complex action of DB04552 on ClC-Ka . The model predicts that ClC-Ka possesses two DB04552 binding sites : when only one site is occupied , DB04552 increases ClC-Ka currents , whereas the occupation of both binding sites leads to channel block . Q8N0V5 V ( Mgat5 ) -mediated N-glycosylation negatively regulates Th1 cytokine production by T cells . The differentiation of naive P01730 (+) T cells into either proinflammatory Th1 or proallergic Th2 cells strongly influences autoimmunity , allergy , and tumor immune surveillance . We previously demonstrated that beta1,6GlcNAc-branched complex-type ( Q8N0V5 V ( Mgat5 ) ) N-glycans on TCR are bound to galectins , an interaction that reduces TCR signaling by opposing agonist-induced TCR clustering at the immune synapse . Mgat5(-/-) mice display late-onset spontaneous autoimmune disease and enhanced resistance to tumor progression and metastasis . In this study we examined the role of beta1,6GlcNAc N-glycan expression in Th1/Th2 cytokine production and differentiation . beta1,6GlcNAc N-glycan expression is enhanced by TCR stimulation independent of cell division and declines at the end of the stimulation cycle . DB00075 -activated splenocytes and naive T cells from Mgat5(-/-) mice produce more P01579 and less P05112 compared with wild-type cells , the latter resulting in the loss of P05112 -dependent down-regulation of IL-4Ralpha . DB02034 , an inhibitor of Q16706 , blocked beta1,6GlcNAc N-glycan expression and caused a similar increase in P01579 production by T cells from humans and mice , but no additional enhancement in Mgat5(-/-) T cells . Mgat5 deficiency did not alter P01579 / P05112 production by polarized Th1 cells , but caused an approximately 10-fold increase in P01579 production by polarized Th2 cells . These data indicate that negative regulation of TCR signaling by beta1,6GlcNAc N-glycans promotes development of Th2 over Th1 responses , enhances polarization of Th2 cells , and suggests a mechanism for the increased autoimmune disease susceptibility observed in Mgat5(-/-) mice . Inhibition of chemokine ( CXC motif ) ligand 12/chemokine ( CXC motif ) receptor 4 axis ( P48061 / P61073 ) -mediated cell migration by targeting mammalian target of rapamycin ( P42345 ) pathway in human gastric carcinoma cells . P48061 / P61073 plays an important role in metastasis of gastric carcinoma . DB00877 has been reported to inhibit migration of gastric cancer cells . However , the role of P42345 pathway in P48061 / P61073 -mediated cell migration and the potential of drugs targeting PI3K/ P42345 pathway remains unelucidated . We found that P48061 activated PI3K/Akt/ P42345 pathway in MKN-45 cells . Stimulating CHO- P04264 cells expressing pEGFP-C1- O43739 -PH fusion protein with P48061 resulted in generation of phosphatidylinositol (3,4,5)-triphosphate , which provided direct evidence of activating PI3K by P48061 . Down-regulation of p110β by siRNA but not p110α blocked phosphorylation of Akt and P23443 induced by P48061 . Consistently , p110β-specific inhibitor blocked the P48061 -activated PI3K/Akt/ P42345 pathway . Moreover , P61073 immunoprecipitated by anti-p110β antibody increased after P48061 stimulation and G(i) protein inhibitor pertussis toxin abrogated P48061 -induced activation of PI3K . Further studies demonstrated that inhibitors targeting the PI3K/ P42345 pathway significantly blocked the chemotactic responses of MKN-45 cells triggered by P48061 , which might be attributed primarily to inhibition of mTORC1 and related to prevention of F-actin reorganization as well as down-regulation of active RhoA , Rac1 , and Cdc42 . Furthermore , rapamycin inhibited the secretion of P48061 and the expression of P61073 , which might form a positive feedback loop to further abolish upstream signaling leading to cell migration . Finally , we found cells expressing high levels of cxcl12 were sensitive to rapamycin in its activity inhibiting migration as well as proliferation . In summary , we found that the P42345 pathway played an important role in P48061 / P61073 -mediated cell migration and proposed that drugs targeting the P42345 pathway may be used for the therapy of metastatic gastric cancer expressing high levels of cxcl12 . Potential opposite roles of the extracellular signal-regulated kinase ( P29323 ) pathway in autism spectrum and bipolar disorders . Signal transduction from the synapse to the nucleus subsequently involves transient increases in synaptic Ca2+ , activation of P62158 kinases , activation of the GTPase Ras , activation of the P29323 mitogen-activated protein kinase pathway , and finally GSK3 inhibition and CREB-activation . Genetic studies in autism have identified mutations and copy number variations in a number of genes involved in this synapse to nucleus signaling path . In particular , a gain of function mutation in the Q13936 gene , deletions and disruption of the Q96PV0 gene , a copy number variation encompassing the P27361 gene and a duplication of P62258 indicate that in a subset of autism patients the P29323 cascade is inappropriately activated . Predicted functional consequences of this hyperactivation would be an increase in complexity of the dendritic tree , and via inhibition of GSK3 , a delayed circadian phase . The latter effect indeed fits the frequent sleep disturbances observed in autistic patients . Interestingly , the sleep disturbances in bipolar disorder patients are frequently characterized as phase advanced . A selective evaluation of genetic mutations in bipolar patients indicates that the activity of the P29323 cascade , at least in a subset of patients , presumably is hypoactive . Thus , with respect to the P29323 pathway , autism and bipolar disorder seem each other 's counter pole . Q07973 as a potential target for cancer therapy . Increasing evidence has accumulated to suggest that vitamin D may reduce the risk of cancer through its biologically active metabolite , DB00136 , which inhibits proliferation and angiogenesis , induces differentiation and apoptosis , and regulates many other cellular functions . Thus , it is plausible to assume that rapid clearance of DB00136 by highly expressed Q07973 could interrupt the normal physiology of cells and might be one cause of cancer initiation and progression . In fact , enhancement of Q07973 expression has been reported in literature for many cancers . Based on these findings , Q07973 -specific inhibitors and vitamin D analogs which are resistant to Q07973 -dependent catabolism might be useful for cancer treatment . Q07973 -specific inhibitor VID400 , which is an azole compound , markedly enhanced and prolonged the antiproliferative activity of DB00136 in the human keratinocytes . Likewise , Q07973 -resistant analogs such as 2α-(3-hydroxypropoxy)- DB00136 ( O2C3 ) and its P06681 -epimer ED-71 ( DB05295 ) , and 19nor- 2α-(3-hydroxypropyl)- DB00136 ( MART-10 ) showed potent biological effects . Our in vivo studies using rats revealed that MART-10 had a low calcemic effect , which is a suitable property as an anticancer drug . Much lower affinity of MART-10 for vitamin D binding protein ( DBP ) as compared with DB00136 may be related to its more potent cellular activities . Based on these results , we conclude that ( 1 ) high affinity for P11473 , ( 2 ) resistance to Q07973 -dependent catabolism , ( 3 ) low affinity for DBP , and ( 4 ) low calcemic effect may be required for designing potent vitamin D analogs for cancer treatment . 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 . DB01780 signaling reveals 14-3-3 protein function as a novel step in left-right patterning during amphibian embryogenesis . To gain insight into the molecular mechanisms underlying the control of morphogenetic signals by H+ flux during embryogenesis , we tested DB01780 -A ( FC ) , a compound produced by the fungus Fusicoccum amygdali Del . In plant cells , FC complexes with 14-3-3 proteins to activate H+ pumping across the plasma membrane . It has long been thought that FC acts on higher plants only ; here , we show that exposing frog embryos to FC during early development specifically results in randomization of the asymmetry of the left-right ( LR ) axis ( heterotaxia ) . Biochemical and molecular-genetic evidence is presented that 14-3-3-family proteins are an obligate component of Xenopus FC receptors and that perturbation of 14-3-3 protein function results in heterotaxia . The subcellular localization of 14-3-3 mRNAs and proteins reveals novel cytoplasmic destinations , and a left-right asymmetry at the first cell division . Using gain-of-function and loss-of-function experiments , we show that P62258 protein is likely to be an endogenous and extremely early aspect of LR patterning . These data highlight a striking conservation of signaling pathways across kingdoms , suggest common mechanisms of polarity establishment between C. elegans and vertebrate embryos , and uncover a novel entry point into the pathway of left-right asymmetry determination . 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 . DB06809 and P27487 modulate mobilization , engraftment , and survival of hematopoietic stem and progenitor cells mediated by the P48061 / P48061 - P61073 axis . The chemokine stromal cell-derived factor-1 ( P48061 / P48061 ) and its receptor , P61073 , are involved in a number of facets of the regulation of hematopoiesis at the level of hematopoietic stem ( HSCs ) and progenitor ( HPCs ) cells . Modulation of this ligand-receptor interaction may be of clinical utility . We now report that : ( 1 ) the CC chemokine , macrophage inflammatory protein-1alpha ( MIP-1alpha/ P10147 ) synergizes with DB06809 ( an antagonist of the binding of P48061 / P48061 to P61073 ) to rapidly mobilize HPCs to the blood of mice ; moreover , the combination of granulocyte colony-stimulating factor ( DB00099 ) with DB06809 and MIP-1alpha/ P10147 , given in a specific sequence , mobilizes the greatest number of HPCs compared to any combination of two of these mobilizing agents ; ( 2 ) pretreatment of recipient mice with Diprotin A , an inhibitor of P27487 /Dipeptidylpeptidase IV ( DPPIV ) , enhances the competitive HSCs repopulating capacity of untreated donor cells ; ( 3 ) the survival-enhancing effects of P48061 / P48061 on HPCs subjected in vitro to delayed addition of growth factors ( GFs ) are mediated in part through the cell cycle-related proteins P38936 (cip1/waf1) ( as assessed using P38936 (cip1/waf1) -/- and +/+ mice ) and Mad2 ( using Mad2 +/- and +/+ mice ) ; and ( 4 ) deletion of P27487 /DPPIV on mouse bone marrow cells increases the survival-enhancing effects of P48061 / P48061 on HPCs . These results demonstrate the means to increase the mobilization of HPCs , the engrafting capability of HSCs , and responsiveness of HPCs to the survival-enhancing activity of P48061 / P48061 , effects that may be of practical value . P08473 -blocking agent thiorphan affects cell growth and differentiation in long term culture of mouse bone marrow . P08473 -blocking agent thiorphan was added to long-term cultures of mouse bone marrow cells at the time of culture initiation ( time 0 ) or 2 weeks thereafter , when the stromal layer appears . Cellularity , cell morphology ( in cytospin smears ) and the yield of granulocyte-macrophage progenitor cells ( GM- Q15814 assay in agar ) were recorded . Low concentrations of thiorphan accelerated recovery of the cultures after an initial drop of the cell count . Expansion and maturation of the granulocytic lineage was promoted , with parallel decline of the GM- Q15814 yield . DB08626 probably interfered with the activity of enkephalinase ( endopeptidase 24.11 ) in the cultures . That enzyme is the CD10 surface marker ( P08473 ) of lymphoid , myeloid and stromal elements . Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 [ ( R ) -(-)-2-(4-bromo-2-fluorobenzyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-4-spiro-3'-pyrrolidine-1,2',3,5'-tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K(i) = 10(-)(10) M ) 2000-fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R- and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP(+) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP(+) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP(+) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and/or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4-bromo-2-fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) .

MH_train_1178

MH_train_1178

MH_train_1178

interacts_with DB01030?

multiple_choice

DB03496 induces mitochondrial-mediated apoptosis in murine glioma GL261 cells via release of cytochrome c and apoptosis inducing factor . Glioblastoma ( GBM ) remains one of the most challenging solid cancers to treat due to its highly proliferative , angiogenic and invasive nature . The small molecule CDK inhibitor , flavopiridol , has demonstrated antitumor activity in human xenograft models and is currently in clinical trials showing efficacy in patients with advanced disease . We have developed an experimental animal model using the murine glioma GL261 cells as a novel in vivo system to screen potential therapeutic agents for GBM . Results of in vitro testing demonstrate that flavopiridol has several relevant clinical characteristics such as its ability to : 1. inhibit cell growth ; 2. inhibit cell migration ; 3. decrease expression of cyclin D1 , P11802 and P38936 ; 4. induce apoptosis in cells with high levels of p27 expression ; and 5. decrease the expression of the anti-apoptotic protein Bcl-2 . The mechanism by which flavopiridol induces apoptosis is mitochondrial-mediated . We demonstrate by electron microscopy and immunohistochemistry that drug treatment induces mitochondrial damage that was accompanied by the release of cytochrome c into the cytosol together with the translocation of apoptosis inducing factor ( O95831 ) into the nucleus . This finding in murine glioma cells differs from the mechanism of flavopiridolinduced cell death reported by us for human glioma cells ( Alonso et al. , Mol Cancer Ther 2003 ; 2:139 ) where drug treatment induced a caspase- and cytochrome c-independent pathway in the absence of detectable damage to mitochondria . In apoptotic human glioma cells only translocation of O95831 into the nucleus occurred . Thus , the same drug kills different types of glioma cells by different mitochondrial-dependent pathways . The monoclonal antibody to cytotoxic T lymphocyte antigen 4 , ipilimumab ( DB06186 ) , a novel treatment strategy in cancer management . BACKGROUND : Cytotoxic T lymphocyte antigen 4 ( P16410 ) is an inhibitory regulator of the T cell immune response against tumor cells . DB06186 ( DB06186 ) is a monoclonal antibody directed against P16410 . OBJECTIVE : To describe the basic mechanism of ipilimumab and discuss data available to date with regards to its safety and efficacy profile . METHODS : Data from clinical trials including abstracts were reviewed using the PubMed Database as well as the American Society of Clinical Oncology Abstract Database . CONCLUSIONS : P16410 inhibition with a monoclonal antibody is usually well tolerated and has efficacy as a therapeutic agent in a variety of cancers . The most clinically important toxicities have been related to autoimmune events , and guidelines for treatment of these effects are now available . Preliminary results indicate that therapy with ipilimumab leads to durable responses . Pharmacokinetics and pharmacodynamics are different from those of traditional chemotherapy agents . Phase III studies are currently underway for melanoma . Anti-HLA-DR-triggered monocytes mediate in vitro T cell anergy . Monomorphic MHC class II determinants are attractive targets for immunomodulation . HLA-DR ligation on antigen-presenting cells ( APCs ) can dramatically alter their function or induce cell death . In monocytes , HLA-DR triggering diminishes their capacity to stimulate T cell proliferation . To further investigate this monocyte-dependent T cell inhibition , we activated human T cells +/- HLA-DR triggering on APCs and tested whether this can induce T cell anergy . Only anti-HLA-DR , but not anti-proliferative control agent anti- P08575 , could modulate monocytes in primary cultures with stimulated T cells , so that T cells were hyporesponsive during re-stimulation . Cell separation studies demonstrated that HLA-DR ligation on monocytes is sufficient for mediating T cell anergy . Secretion of monokines was severely reduced after primary culture . Monocytes anergized independently of soluble factors . Extracellular signal-regulated kinase ( P29323 ) phosphorylation occurred early with anti-HLA-DR , but late with anti- P08575 antibody . However , P29323 inhibition did not reverse the T cell-anergizing potential of HLA-DR-ligated monocytes implicating other signaling pathways involved in tolerance induction . When analyzing the anergized T cells , they were refractory to exogenous P60568 and characterized by defective secretion of various cytokines . Expression of CD25 , P10747 , intracellular CD3zeta and P16410 was reduced . The hyporesponsive T cells up-regulated cell-cycle inhibitors p27(kip1) and P38936 (cip1) in correlation with human T cell anergy . In contrast , caspase-3 and -8 , known to contribute to T cell proliferation , were equally decreased in anti-HLA-DR- and anti- P08575 -inhibited cultures . In summary , anti-HLA-DR treatment can generate tolerogenic monocytes transmitting T cell anergy that may be exploited for future immunomodulatory strategies to treat immune-mediated disease states . The kinase inhibitor staurosporine induces P55008 arrest at two points : effect on retinoblastoma protein phosphorylation and cyclin-dependent kinase 2 in normal and transformed cells . DB02010 ( ST ) , a protein kinase inhibitor , at a concentration of 20 nM arrests normal diploid fibroblasts 3 h into P55008 ( H. A. Crissman et al. , Proc. Natl. Acad. Sci. USA , 88 : 7580-7584 , 1991 ; K. Abe et al. , Exp. Cell Res. , 192 : 122-127 , 1991 ) . ST ( 2 nM ) induces a new P55008 arrest point at 6 h into P55008 . Partial phosphorylation of the retinoblastoma protein was observed at the 2 nM ST arrest point , whereas the retinoblastoma protein was unphosphorylated or underphosphorylated at the 20 nM arrest point . This correlated with the activity of the cyclin-dependent kinase 2 ( P24941 ) and the phosphorylation of the Thr160 residue of p33CDK2 . The cyclin E and cyclin D1/2 levels were reduced at the 20 nM ST arrest point . In HeLa cells that do not arrest in P55008 in response to 2 or 20 nM ST , the retinoblastoma protein and P24941 phosphorylations and P24941 activity were not affected by ST . These results suggest that ST inhibits one or more P55008 -regulating protein kinases , which lie upstream of P24941 . P11387 is a cofactor for c-Jun in the regulation of epidermal growth factor receptor expression and cancer cell proliferation . P11387 ( Topo I ) is a molecular target for the anticancer agent topotecan in the treatment of small cell lung cancer and ovarian carcinomas . However , the molecular mechanisms by which topotecan treatment inhibits cancer cell proliferation are unclear . We describe here the identification of Topo I as a novel endogenous interaction partner for transcription factor c-Jun . Reciprocal coimmunoprecipitation analysis showed that Topo I and c-Jun interact in transformed human cells in a manner that is dependent on JNK activity . c-Jun target gene epidermal growth factor receptor ( P00533 ) was identified as a novel gene whose expression was specifically inhibited by topotecan . Moreover , Topo I overexpression supported c-Jun-mediated reporter gene activation and both genetic and chemical inhibition of c-Jun converted cells resistant to topotecan-elicited P00533 downregulation . DB01030 -elicited suppression of proliferation was rescued by exogenously expressed P00533 . Furthermore , we demonstrate the cooperation of the JNK-c-Jun pathway , Topo I , and P00533 in the positive regulation of O75794 cell proliferation . Together , these results have identified transcriptional coactivator Topo I as a first endogenous cofactor for c-Jun in the regulation of cell proliferation . In addition , the results of the present study strongly suggest that inhibition of P00533 expression is a novel mechanism by which topotecan inhibits cell proliferation in cancer therapy . Suppression of rat breast cancer metastasis and reduction of primary tumour growth by the small synthetic urokinase inhibitor DB05476 . The serine protease uPA ( urokinase-type plasminogen activator ) and its receptor Q03405 ( CD87 ) are often elevated in malignant tumours , hence , inhibition of this tumour-associated plasminogen activation system provides an attractive target for therapeutic strategies . DB05476 , a derivative of 3-aminophenylalanine in the L-conformation with inhibitory antiproteolytic properties , was tested for its specificity spectrum using specific chromogenic paranitroanilide peptide substrates . The corresponding D-enantiomer of DB05476 was used as a control . The anti-tumour and anti-metastatic ( number of lung foci and weight of the axillary lymph nodes ) properties were studied by subcutaneous administration of DB05476 to Brown Norwegian ( BN ) rats carrying orthotopically transplanted BN472 rat breast tumours . DB05476 selectively inhibited tumour-related proteases from rats and humans such as uPA , plasmin , or thrombin in the sub or low micromolar range . The activity was stereoselective as the D-enantiomer of DB05476 inhibited uPA and plasmin at approximately 70-fold higher Ki values than the active L-form . Chronical administration of the L-enantiomer of WXUK1 impaired primary tumour growth and metastasis of BN472 rat breast cancer in a dose-dependent manner . The minimum inhibitory dosage with maximal effect was between 0.15 and 0.3 mg/kg/day . The inactive D-enatiomer of DB05476 was not active in this respect . Daily treatment with DB05476 for up to 35 days was well tolerated as judged by the unchanged body and organ weight development . In conclusion , our results provide evidence that DB05476 as a single agent inhibits breast tumour growth and metastasis in vivo , and thus is a promising candidate drug to treat human cancer . 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 . 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 . Nuclear factor kappa B inhibition improves conductance artery function in type 2 diabetic mice . BACKGROUND : We previously reported that enhanced nuclear factor kappa B ( NFκB ) activity is responsible for resistance arteries dysfunction in type 2 diabetic mice . METHODS : In this study , we aimed to determine whether augmented NFκB activity also impairs conductance artery ( thoracic aorta ) function in type 2 diabetic mice . We treated type 2 diabetic ( db(-) /db(-) ) and control ( db(-) /db(+) ) mice with two NFκB inhibitors ( dehydroxymethylepoxyquinomicin , 6 mg/kg , twice a week and IKK-NBD peptide , 500 µg/kg/day ) for 4 weeks . RESULTS : As expected , the NFκB inhibition did not affect blood glucose level and body weight . Thoracic aorta vascular endothelium-dependent relaxation ( EDR ) , determined by the wire myograph , was impaired in diabetic mice compared with control and was significantly improved after NFκB inhibition . Interestingly , thoracic EDR was also rescued in db(-) /db(-p50NFκB-/-) and db(-) /db(- P09874 -/-) double knockout mice compared with db(-) /db(-) mice . Similarly , the acute in vitro down regulation of NFκB-p65 using p65 shRNA lentiviral particles in arteries from db(-) /db(-) mice also improved thoracic aorta EDR . Western blot analysis showed that the p65NFκB phosphorylation , cleaved P09874 and P35354 expression were increased in thoracic aorta from diabetic mice , which were restored after NFκB inhibition and in db(-) /db(-p-50NFκB-/-) and db(-) /db(- P09874 -/-) mice . CONCLUSIONS : The present results indicate that in male type 2 diabetic mice , the augmented NFκB activity also impairs conductance artery function through P09874 and P35354 -dependent mechanisms . Discovery of ( 2E ) -3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide ( DB05223 ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 enzyme and COLO 205 cellular IC(50) ) , liver microsomal stability ( t(1/2) ) , cytochrome P450 inhibitory ( 3A4 IC(50) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT-116 , PC-3 , A2780 , MV4-11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials . The overexpression of Q8NEM0 inhibits cell growth through regulating cell cycle-related proteins and activating cytochrome c-caspase 3 signaling in cervical cancer . Q8NEM0 , initially identified as an hTERT repressor , has recently been implicated in mediating DNA damage response and maintaining chromosome integrity . This study is to investigate its potential role in the onset of cervical cancer . In the study , decreased expression of Q8NEM0 was observed in 19 of 31 cases ( 61.3 % ) at mRNA level and 44 of 63 cases ( 69.8 % ) at protein level of cervical tumor tissues compared with the paired nontumor tissues . Reduced Q8NEM0 protein expression was significantly associated with high-tumor grade ( 1 vs. 3 P = 0.013 ; 2 vs. 3 P = 0.047 ) . In addition to inhibit SiHa cell migration and invasion , the overexpression of Q8NEM0 inhibited cervical cancer cells growth through inducing S phase arrest and mitochondrial apoptosis . Further analysis demonstrated cyclinA2/ P24941 , P30307 -cyclinB/ P06493 , and p53/ P38936 pathways were involved in the Q8NEM0 overexpression-induced S phase arrest . Moreover , the overexpression of Q8NEM0 activated mitochondrial apoptosis through regulating several apoptosis-related proteins such as p53 , Bcl-2 , Bax , cytochrome c , caspase-3 , and P09874 . Our findings indicate that downregulated Q8NEM0 correlates with tumor progression in cervical cancer , and Q8NEM0 has an important role in regulating cell growth through regulating the cell cycle and apoptosis . Thus , it may be a crucial tumor suppressor gene and a novel candidate therapeutic target for cervical cancer . Non-toxic dose chidamide synergistically enhances platinum-induced DNA damage responses and apoptosis in Non-Small-Cell lung cancer cells . Combination of low doses of histone deacetylases inhibitors and chemotherapy drugs is considered as one of the most promising strategies to increase the anticancer efficacy . Chidamide is a novel benzamide chemical class of HDAC inhibitor that selectively inhibited Q13547 , 2 , 3 and 10 . We sought to determine whether chidamide may enhance platinum-induced cytotoxicity in NSCLC cells . In this study , the combination of chidamide with carboplatin showed a good synergism on growth inhibition with the mean combination index value as 0.712 and 0.639 in A549 and NCI-H157 cells , respectively . The used concentration of chidamide was non-toxic on cells by itself as low as 0.3μM . All of our experiments were comparisons between combination regimen and single carboplatin regimen in A549 and NCI-H157 cell lines . Phosphorylated histone H2A.X ( γH2A.X ) , a hall marker of DNA damage response , was dramatically increased by the combination treatment . Cell cycle analysis by flow cytometry and phosphorylation level analysis of histone H3 ( Ser10 ) by western blotting showed that combination treatment significantly increased the percentage of G2/M phase of cells . Mitochondrial membrane potential and cleaved- P09874 level analysis indicate that chidamide synergistically enhances carboplatin-induced apoptosis . Additionally , synergistic effects of chidamide were found when it was combined with two other platinum drugs ( cisplatin and oxaliplatin ) . The results suggest that Chidamide in combination with platinum drugs may be a novel therapeutic option for NSCLC . DB01283 . DB01283 is a highly selective and potent cyclo-oxygenase ( P36551 ) -2 inhibitor , with a novel structure that conveys weakly acidic properties and a unique pharmacological profile . It is rapidly absorbed , with a relatively short plasma half-life . In well designed clinical trials of 1-52 weeks ' duration in patients with osteoarthritis ( OA ) or rheumatoid arthritis , the efficacy of oral lumiracoxib 100-400 mg/day in decreasing pain intensity and improving functional status was greater than that with placebo and similar to those with nonselective NSAIDs or celecoxib 200mg once daily . In single- and multiple-dose well designed trials in patients with acute pain associated with primary dysmenorrhoea , dental or orthopaedic surgery or tension-type headache , lumiracoxib 100-800 mg once daily was more effective in relieving acute pain than placebo or controlled-release oxycodone 20 mg , and was at least as effective as selective P35354 inhibitors or nonselective NSAIDs . DB01283 was generally well tolerated in clinical trials , with a similar overall tolerability profile to those of placebo and other P35354 -selective inhibitors . In a large 52-week safety trial in patients with OA , lumiracoxib 400mg once daily had a rate of gastrointestinal ulcer complications that was approximately one-third to one-quarter of that of ibuprofen 800 mg three times daily or naproxen 500 mg twice daily . DB01283 was not associated with an increase in cardiovascular events . DB05101 binding to P00533 prevents the conformational rearrangement required for dimerization . An increasing number of therapeutic antibodies targeting tumors that express the epidermal growth factor receptor ( P00533 ) are in clinical use or late stages of clinical development . Here we investigate the molecular basis for inhibition of P00533 activation by the therapeutic antibody matuzumab ( EMD72000 ) . We describe the X-ray crystal structure of the Fab fragment of matuzumab ( Fab72000 ) in complex with isolated domain III from the extracellular region of P00533 . Fab72000 interacts with an epitope on P00533 that is distinct from the ligand-binding region on domain III and from the cetuximab/Erbitux epitope . DB05101 blocks ligand-induced receptor activation indirectly by sterically preventing the domain rearrangement and local conformational changes that must occur for high-affinity ligand binding and receptor dimerization . Mulberry ( Morus L. ) methionine sulfoxide rreductase gene cloning , sequence analysis , and expression in plant development and stress response . DB00134 sulfoxide reductase plays a regulatory role in plant growth and development , especially in scavenging reactive oxygen species by restoration of the oxidation of methionine in protein . A full-length cDNA sequence encoding methionine sulfoxide reductase ( Q9UBK8 ) from mulberry , which we designated MMSR , was cloned based on mulberry expressed sequence tags ( ESTs ) . Sequence analysis showed that the MMSR is 810 bp long , encoding 194 amino acids with a predicted molecular weight of 21.6 kDa and an isoelectric point of 6.78 . The expression level of the MMSR gene under conditions of drought and salt stresses was quantified by qRT-PCR . The results show that the expression level changed significantly under the stress conditions compared to the normal growth environment . It helps us to get a better understanding of the molecular basis for signal transduction mechanisms underlying the stress response in mulberry . Involvement of c-myc-regulated genes in hepatocellular carcinoma related to genotype-C hepatitis B virus . PURPOSE : The purpose of this study was to elucidate the molecular basis of hepatocellular carcinoma ( HCC ) caused by genotype-C hepatitis B virus ( HBV ) . METHODS : We compared molecular profiles of 15 HCCs and five non-tumorous livers , all of which were associated with genotype-C HBV infection , using DNA microarray technology . RESULTS : Our supervised learning identified 237 genes whose expression differed between HCCs and non-tumorous livers . This result was validated by a false discovery rate of 0 % . Levels of expression of 35 and 202 genes were higher and lower , respectively , in HCCs than in non-tumorous livers . Among the 237 genes , we highlighted the top 35 upregulated and top 35 down regulated genes in tumor . Interestingly , when overlapping genes were excluded , 12 ( e.g. , P15531 -H2 , P33993 , P09874 , Q04917 , P04792 , and P43246 ) of the top 34 upregulated genes and five ( e.g. , P04731 and P25713 ) of the top 33 downregulated genes were c-myc-regulated genes . The microarray data for five randomly selected genes ( P33993 , P68036 , P62937 , P48061 , and P00966 ) were confirmed by quantitative real-time reverse transcription-polymerase chain reaction . CONCLUSIONS : Our results indicate that many c-myc-regulated genes are involved in genotype-C-HBV-related HCC , suggesting that c-myc is related to the hepatocarcinogenic activity of genotype-C HBV . P01375 -alpha -mediated protein kinases in regulation of scavenger receptor and foam cell formation on macrophage . We previously reported tumor necrosis factor-alpha ( P01375 ) modulates transcriptional and post-transcriptional down-regulation of macrophage scavenger receptor ( Q9UBK8 ) ( Hsu , H. Y. , Nicholson , A. C. , and Hajjar , D. P. ( 1996 ) J. Biol. Chem. 271 , 7767-7773 ) ; however , P01375 -mediated signaling mechanisms are unknown . Here , we demonstrate that ligation of P01375 receptor stimulates activity of P38936 -activated protein kinase ( PAK ) and mitogen-activated protein kinases ( MAPK ) as follows : P29323 , JNK , and p38 in murine macrophage J774A.1 cells . Upon activation of protein kinases ( PK ) , P01375 rapidly increases Q9UBK8 message and protein ; later it markedly reduces Q9UBK8 expression . Studies using PK inhibitors and dominant negative constructs demonstrate phosphatidylinositol 3-kinase/Rac1/PAK/JNK and phosphatidylinositol 3-kinase/Rac1/PAK/p38 pathways contribute to important roles in the late stage of P01375 down-regulation of Q9UBK8 expression and taking up of OxLDL . Alternatively , the PKC/ Q02750 / P29323 pathway in the early stage plays a significant role in up-regulation of the Q9UBK8 gene . By using anti- P19438 agonist antibody , we further confirm P19438 -mediated MAPK in regulation of Q9UBK8 . Furthermore , in Q9UBK8 gene promoter-driven luciferase reporter assays with P01375 , PKC activator increases , but antioxidant DB06151 , PK inhibitors , and dominant negative constructs decrease luciferase activity in Q9UBK8 gene promoter-transfected cells . Our current results show the first evidence of crucial roles for P01375 -mediated MAPK pathways in the transcriptional regulation of Q9UBK8 gene and increase Q9UBK8 expression ; in contrast , with P01375 longer treatment the pathways down-regulate Q9UBK8 and foam cell formation probably via post-transcriptional process . Plasma biomarkers correlating with clinical outcome in a phase II study of sorafenib in advanced NSCLC . We investigated the relationship between plasma protein biomarker concentrations and clinical outcomes in 52 patients with relapsed/refractory advanced non-small cell lung cancer ( NSCLC ) treated with 400~mg bid sorafenib in a phase II trial . Blood samples were collected at baseline , on day 15 of cycle 1 ( C1D15 ) , and on day 1 of cycle 3 ( C3D1 ) , and plasma concentrations of total P15692 , P15692 -165 , soluble ( s ) P35968 , DB00102 , sPDGFR-β , sEGFR , sHER-2 , uPA , P05121 , Q03405 , P01033 , and circulating Ras P38936 were assayed by ELISA . Elevated baseline P15692 , P15692 -165 , DB00102 , Ras P38936 , and P01033 concentrations were associated with poorer patient outcomes ( shorter overall survival [ OS ] and/or progression-free survival [ PFS ] ) . During treatment , the mean concentrations of sVEGFR-2 , DB00102 , sPDGFR-β , P01033 , Q03405 , and P05121 decreased , while the mean sEGFR concentration increased . Increases in P15692 , P15692 -165 , DB00102 , and P01033 during treatment were associated with better outcomes ( longer OS and/or PFS ) , whereas increases in plasma Ras P38936 during treatment were associated with shorter PFS . The associations between baseline concentrations and/or pharmacodynamic changes in plasma proteins and clinical outcomes in NSCLC patients treated with sorafenib suggest that these biomarkers may have a prognostic role and/or predict the efficacy of sorafenib in patients with NSCLC .

MH_train_1179

MH_train_1179

MH_train_1179

interacts_with DB00603?

multiple_choice

Binding affinities for sulfonamide inhibitors with matrix metalloproteinase-2 using a linear response method . Due to their involvement in many pathological conditions , matrix metalloproteinases ( MMPs ) , are very attractive therapeutic targets . Our study focuses on one of them , P08253 , which is involved in tumor progression and metastasis . Recently , the solution structure of the catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) was published by Feng et al . Using the Hanessian group published binding affinity data and the structure published by Feng as a basis , we have built a binding affinity model by targeting the S(2) ' pocket of the enzyme with a set of nine alpha-N-sulfonylamino hydroxamic acid derivatives . Two binding geometries of each ligand have been generated corresponding to two binding modes denoted A and B , respectively , of which the first one is targeting the S(2) ' pocket and the second one the S(1) pocket . For the binding affinity model developed for mode A the computed activities show a rmsd of 0.583 kcal/mol as compared with the experimental data , and a correlation coefficient r(2) of 0.779 , while in the case of the binding mode B a rmsd of 0.834 kcal/mol and correlation coefficient r(2) of 0.500 , respectively , were obtained . In conclusion , our data suggest a higher probability for the DB00120 (76) gated S(2) ' open form pocket to accommodate the substituent alpha versus the wide solvent exposed S(1) subsite , probability which some research groups could have overlooked due to extensive use in their calculations of non revealing S(2) ' pocket open state crystallographic structures instead of NMR ones . 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 . Proliferation of endothelial cell on polytetrafluoroethylene vascular graft materials carried P15692 gene plasmid . OBJECTIVE : To investigate whether vascular endothelial growth factor ( P15692 ) gene plasmid carried by polytetrafluoroethylene ( PTFE ) vascular graft materials could transfect endothelial cells ( ECs ) and promote their growth . METHODS : PTFE vascular graft materials carried with pCDI-hVEGF(121) , pCDI or pEGFP were incubated in DB03754 -buffer solution and the values of optical density of 260 nm at different time were plotted , then the DNA controlled release curve was made . ECs derived from human umbilical vein were seeded on the pCDI-hVEGF(121)/pCDI/pEGFP-PTFE materials or tissue culture plates , ECs numbers were counted and P15692 protein concentrations at different time were measured by enzyme-linked immunoadsorbent assay method . Green fluorescent protein ( GFP ) expression in ECs on pEGFP-PTFE materials was examined with fluorescence microscopy . RESULTS : The controlled release curve showed that the gene released from PTFE materials was rapid within 8 h , then slowed down and that the gene released continuously even after 72 h . At 24 , 72 and 120 h , ECs number and proliferation rate of pCDI-hVEGF(121)-PTFE materials were higher than those of pCDI or pEGFP-PTFE materials ( P < 0.05 ) . P15692 protein concentration of pCDI-hVEGF(121)-PTFE materials was higher than that of pCDI or pEGFP-PTFE materials at 6 , 24 , 72 and 120 h ( P < 0.01 ) . GFP expression in ECs on the pEGFP-PTFE materials could be detected by fluorescence microscopy . CONCLUSION : PTFE graft can be used as a carrier of P15692 gene plasmid , P15692 gene carried by PTFE can transfect ECs and promote ECs growth . P23582 ( P09543 ) is the major natriuretic peptide in human cerebrospinal fluid . In order to investigate whether P23582 ( P09543 ) is present in human cerebrospinal fluid ( P04141 ) , we measured P09543 -like immunoreactivity ( -LI ) in human P04141 by specific radioimmunoassay ( RIA ) for P09543 . We also measured atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) concentrations in human P04141 . P01160 -LI , DB04899 -LI , and P09543 -LI concentrations of P04141 collected from fifteen patients without neurological disorders were 0.20 +/- 0.13 , 0.27 +/- 0.10 , and 2.13 +/- 0.27 fmol/ml ( mean +/- S.D. ) , respectively . In fifteen patients with neurological disorders , P01160 -LI , DB04899 -LI , and P09543 -LI concentrations in P04141 were 0.21 +/- 0.18 , 0.33 +/- 0.19 , and 2.09 +/- 0.82 fmol/ml , respectively . Although P01160 -LI and DB04899 -LI concentrations in plasma were much higher than those in P04141 , P09543 -LI was undetectable in plasma ( less than 0.2 fmol/ml ) . These results demonstrate that three natriuretic peptides are present in P04141 and that P09543 is the major natriuretic peptide in human P04141 . These results suggest that P09543 in P04141 is originated from and play important roles in the central nervous system . 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 . 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 review of treatment with mepolizumab , an anti- P05113 mAb , in hypereosinophilic syndromes and asthma . The hypereosinophilic syndromes ( HESs ) are a heterogeneous group of diseases characterized by peripheral blood eosinophilia with end-organ damage and varying in severity from nonspecific symptoms to life-threatening . Treatment objectives are a safe reduction of blood and tissue eosinophil levels and prevention of eosinophil-mediated tissue damage . Current treatment of patients with HESs , who lack the Q7L523 -like 1-platelet-derived growth factor receptor alpha ( Q6UN15 - P16234 ) fusion gene , is mainly systemic corticosteroid therapy . Eosinophil development from hematopoietic progenitor cells is regulated by P05113 , which influences maturation , differentiation , mobilization , activation , and survival . Consequently , inhibiting P05113 is a logical therapeutic objective for patients with HESs or selected patients with asthma . DB06612 is a fully humanized anti- P05113 monoclonal IgG(1) antibody that binds to free P05113 with high affinity and specificity to prevent P05113 from associating with the P05113 receptor complex alpha-chain on the surface of eosinophils . In clinical trials with patients with HESs , mepolizumab reduced blood eosinophil counts and the maintenance corticosteroid dose and had no major safety concerns . DB06612 reduced airway and blood eosinophils and prevented asthma exacerbations . Thus , mepolizumab may be effective for long-term treatment of patients with selected eosinophilic disorders . 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 . Aldo-keto reductase 1 family P33681 is the gene induced in response to oxidative stress in the livers of Long-Evans Cinnamon rats . The Long-Evans Cinnamon ( LEC ) rat strain ( Atp7b m/m ) , which accumulates copper in the liver due to mutations in the Atp7b gene , is a useful model for investigating the relationship between oxidative stress and hepatocarcinogenesis . To determine the effect of this mutation on oxidative stress marker genes , we performed oligonucleotide array analysis ( Affymetrix ) , and compared the results in Atp7b m/m rats with those of a sibling line with the Atp7b w/w genotype . We focused our studies on the expression of the aldo-keto reductase 1 family P33681 ( AKR1B7 ) -like protein gene , since this gene codes for reductase enzymes involved in the detoxification of oxidizing compounds ( e.g. , aldehydes ) and was differentially expressed in Atp7b m/m and Atp7b w/w rat liver . Akr1B7 mRNA expression was significantly increased in comparison with the expression of 4 other known oxidative stress responsive genes , haem-oxygenase-1 ( P09601 ) , thioredoxin ( P10599 ) , aldehyde reductase ( P14550 ) , and glucose-6-phosphate dehydrogenase ( G6PDH ) . By searching binding motifs , five nuclear factor kappa B ( NF-kappaB ) binding sites were located in the 5'-upstream region of the akr1b7 gene . Transient co-transfection with both I-kappaBalpha and the Akr1b7 6 kb promoter ( P80511 .0-AKR-Luc ) inhibited luciferase activity of P80511 .0-AKR-Luc in HepG2 cells . Cuprous ion however did not affect the transcription activity induced by P80511 .0-AKR-luc . Gel-shift assay showed that the DNA binding activity of NF-kappaB increased in the livers of LEC rats , suggesting that the oxidative stress is mediated through NF-kappaB . The results indicate conclusively that in LEC rat liver , akr1b7 might be up-regulated by oxidative stress mediated through NF-kappaB , but not that mediated directly by copper . Prevention of airway inflammation with topical cream containing imiquimod and small interfering RNA for natriuretic peptide receptor . BACKGROUND : Asthma is a complex disease , characterized by reversible airway obstruction , hyperresponsiveness and chronic inflammation . Principle pharmacologic treatments for asthma include bronchodilating beta2-agonists and anti-inflammatory glucocorticosteroids ; but these agents do not target the main cause of the disease , the generation of pathogenic Th2 cells . We previously reported reduction in allergic inflammation in mice deficient in the P01160 receptor NPRA . Here we determined whether siRNA for natriuretic peptide receptor A ( siNPRA ) protected against asthma when administered transdermally . METHODS : Imiquimod cream mixed with chitosan nanoparticles containing either siRNA green indicator ( siGLO ) or siNPRA was applied to the skin of mice . Delivery of siGLO was confirmed by fluorescence microscopy . The anti-inflammatory activity of transdermal siNPRA was tested in OVA-sensitized mice by measuring airway hyperresponsiveness , eosinophilia , lung histopathology and pro-inflammatory cytokines . RESULTS : SiGLO appearing in the lung proved the feasibility of transdermal delivery . In a mouse asthma model , BALB/c mice treated with imiquimod cream containing siNPRA chitosan nanoparticles showed significantly reduced airway hyperresponsiveness , eosinophilia , lung histopathology and pro-inflammatory cytokines P05112 and P05113 in lung homogenates compared to controls . CONCLUSION : These results demonstrate that topical cream containing imiquimod and siNPRA nanoparticles exerts an anti-inflammatory effect and may provide a new and simple therapy for asthma . 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 . DB00174 Synthetase Deficiency : New Inborn Errors of Metabolism . BACKGROUND : DB00174 synthetase deficiency ( P51689 ) is a newly identified neurometabolic disorder characterized by severe congenital microcephaly , severe global developmental delay , intractable seizure disorder , and spastic quadriplegia . Brain Q9BWK5 showed brain atrophy , delayed myelination , and simplified gyriform pattern . METHODS : We report P51689 deficiency in a 2- and 4-year-old sibling . On them , we described clinical , biochemical , and molecular findings , and we compared our results with previously reported cases . RESULTS : We identified a homozygous novel missense mutation in P08243 gene in both probands and we demonstrated low P04141 and plasma asparagine in both patients . CONCLUSIONS : Clinicians should suspect P51689 deficiency in any newborn presented with severe congenital microcephaly followed by severe epileptic encephalopathy and global developmental delay . P04141 asparagine level is low in this disorder while plasma may be low . P06401 membrane component 1 as the mediator of the inhibitory effect of progestins on cytokine-induced matrix metalloproteinase 9 activity in vitro . Progesterone ( P4 ) and the progestin , 17α-hydroxyprogesterone caproate , are clinically used to prevent preterm births ( PTBs ) ; however , their mechanism of action remains unclear . Cytokine-induced matrix metalloproteinase 9 ( P14780 ) activity plays a key role in preterm premature rupture of the membranes and PTB . We demonstrated that the primary chorion cells and the HTR8/SVneo cells ( cytotrophoblast cell line ) do not express the classical progesterone receptor ( P06401 ) but instead a novel progesterone receptor , progesterone receptor membrane component 1 ( O00264 ) , whose role remains unclear . Using HTR8/SVneo cells in culture , we further demonstrated that 6 hours pretreatment with medroxyprogesterone acetate ( DB00603 ) and dexamethasone ( DB00514 ) but not P4 or 17α-hydroxyprogesterone hexanoate significantly attenuated tumor necrosis factor α-induced P14780 activity after a 24-hour incubation period . The inhibitory effect of DB00603 , but not DB00514 , was attenuated when O00264 expression was successfully reduced by O00264 small interfering RNA . Our findings highlight a possible novel role of O00264 in mediating the effects of DB00603 and in modulating cytokine-induced P14780 activity in cytotrophoblast cells in vitro . Tp53-associated growth arrest and DNA damage repair gene expression is attenuated in mammary epithelial cells of rats fed whey proteins . Dietary protection from mammary cancer is likely coordinated through multiple signaling pathways , based on the known heterogeneity of the disease and the distinct origins of mammary tumor cells . The present study examined the modulatory effects of dietary intake of whey protein hydrolysate ( WPH ) relative to casein ( CAS ) , on mammary epithelial cell resistance to endogenous DNA damage using Tp53 gene expression and signaling as a read-out , and on systemic proapoptotic and immune surveillance activity , in young adult female Sprague-Dawley rats . Rats were fed AIN-93G diets made with CAS or WPH as the sole protein source beginning at gestation d 4 . At postnatal day ( P01160 ) 50 , mammary glands of rats fed WPH had lower levels of activated Tp53 and p38 mitogen-activated protein kinase proteins , and reduced transcript levels for Tp53-associated DNA damage repair , growth arrest , and proapoptotic genes than those of CAS-fed rats . Serum from WPH-fed rats had greater apoptotic activity in MCF-7 tumor cells than that from rats fed CAS . Serum levels of monocyte chemoattractant protein ( MCP ) -1 were higher in WPH- than in CAS-fed rats . MCF-7 cells treated with CAS serum + recombinant rat P13500 had apoptotic activity and Tp53 and P38936 gene expression levels comparable to those treated with WPH serum or recombinant P13500 . Results indicate that mammary glands of rats fed a WPH diet are more protected from endogenous DNA damage than are those of CAS-fed rats , and identify P13500 as a potential serum biomarker for the positive effects of healthy diets . DB08836 treatment ameliorates autoimmune myocarditis associated with enhanced cardiomyocyte thioredoxin expression . OBJECTIVE : P10599 ( TRX ) is a redox regulatory protein that protects cells from various stresses . P12821 ( P12821 ) inhibitor was reported to enhance endogenous antioxidant enzyme activities . This study was carried out to investigate whether temocapril , a novel non-sulfhydryl-containing P12821 inhibitor , reduces the severity of myocarditis via redox regulation mechanisms involving TRX . METHODS AND RESULTS : In normal rat myocytes in vitro and in vivo , Western blot showed that temocapril enhanced cytosolic redox regulatory protein TRX expression , but that neither mitochondrial TRX2 nor antioxidant enzymes , such as copper-zinc superoxide dismutase ( Cu/Zn-SOD ) or manganese superoxide dismutase ( Mn-SOD ) expression , was up-regulated by the preconditioning treatment . In rats with experimental autoimmune myocarditis ( EAM ) , the severity of myocarditis and the protein carbonyl contents were less increased in temocapril treatment ( 10 mg/kg/day , orally ) from day 1 to day 21 , but not in temocapril treatment from day 15 to day 21 . An immunohistochemical study showed that TRX stain was enhanced in infiltrating inflammatory cells and in damaged myocytes . Considering the characteristics of this model that myocardial inflammation begins around day 15 and increases until day 21 , temocapril treatment for 3 weeks might be thought of as a preconditioning treatment . CONCLUSIONS : The results suggest that TRX and the redox state modified by TRX may play a crucial role in the pathophysiology of EAM . DB08836 ameliorates myocarditis associated with inducing TRX up-regulation in a preconditioning manner , although the mechanism of TRX up-regulation by temocapril remains to be elucidated . 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 . 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 . Monoclonal antibodies targeting P01584 reduce biomarkers of atherosclerosis in vitro and inhibit atherosclerotic plaque formation in P02649 -deficient mice . OBJECTIVE : Atherosclerosis is a condition that is increasingly contributing to worldwide mortality through complications such as stroke and myocardial infarction . IL-1β plays multiple direct , local roles in the formation and stability of the atheroma by eliciting the production of additional cytokines and proteolytic enzymes from macrophages , endothelial cells ( EC ) and smooth muscle cells ( SMC ) . We therefore tested whether an anti-IL-1β antibody , DB06062 , might inhibit the secretion of pro-atherogenic cytokines from macrophages in vitro and affect a positive outcome in the P02649 -deficient mouse ( ApoE(-/-) ) model of atherosclerosis in vivo . METHODS AND RESULTS : In an in vitro co-culture model , DB06062 inhibited macrophage-induced secretion of key atherogenic cytokines from EC and SMC , including P05231 , P10145 , P13500 and TNFα . The release of degradative enzymes , such as the matrix metalloproteinases P08254 and P14780 , was also decreased by DB06062 . In addition , DB06062 inhibited the secretion of P13232 from EC and P05112 from SMC , cytokines not previously reported to be driven by IL-1β in this context . In vivo , XMA052 MG1K , a chimeric murine version of DB06062 , inhibited the formation of atherosclerotic lesions in the ApoE(-/-) model at all three doses tested . This effect was comparable to that reported for complete genetic ablation of IL-1β or IL-1R1 on an ApoE(-/-) background and was associated with decreases in plasma non-HDL/HDL cholesterol ratio and plaque lipid content and macrophage infiltration . CONCLUSIONS : These results demonstrate for the first time that an antibody targeting IL-1β can inhibit the progression of atherosclerosis in vivo , highlighting the importance of this key cytokine in cardiovascular disease . Involvement of protein kinase C-dependent mitogen-activated protein kinase Q8TCB0 /42 signaling pathway for cross-talk between estradiol and transforming growth factor-beta3 in increasing basic fibroblast growth factor in folliculostellate cells . We have recently shown that TGF-beta3 , in the presence of estradiol , increases the release of basic fibroblast growth factor ( P09038 ) from folliculostellate ( FS ) cells in the pituitary . We determined the interactive effects of TGF-beta3 and estradiol on P09038 production and release from FS cells , and the role of the MAPK pathway in TGF-beta3 and estradiol interaction . We found that TGF-beta3 and estradiol alone moderately increased cell content and release of P09038 from FS cells ; but together , they markedly increased the peptide . Estradiol and TGF-beta3 alone moderately activated MAPK Q8TCB0 /42 ; together they produced marked activation of MAPK Q8TCB0 /42 . Pretreatment of FS cells with an MAPK kinase 1/2 inhibitor or with protein kinase C inhibitors suppressed the activation of MAPK Q8TCB0 /42 , P09038 release , and protein level increases , all of which were induced by TGF-beta3 and estradiol . Estradiol and TGF-beta3 , either alone or in combination , increased the levels of active Ras . Furthermore , P09038 induction by TGF-beta3 and estradiol was blocked by overexpression of Ras N17 , a dominant negative mutant of Ras P38936 . P03372 blocker DB00947 failed to prevent estrogen 's and TGF-beta3 's effects on P09038 . These data suggest that an estradiol receptor-independent protein kinase C- activated Ras-dependent MAPK pathway is involved in the cross-talk between TGF-beta3 and estradiol to increase P09038 production and/or release from FS cells . Reversal of LPS-induced immobility in mice by green tea polyphenols : possible P35354 mechanism . An endotoxin ( lipopolysaccharide , LPS ) is known to activate the hypothalamo-pituitary adrenocortical axis , as well as norepinephrine and indolamine metabolism . Systemically administered LPS produces depression in the forced swimming-induced despair behaviour model in mice . The present study was designed to investigate the effect of green tea extract ( GTE ) on LPS-induced despair behaviour and to explore the mechanism involved in modulation of LPS-induced immobility by GTE . GTE ( 10-100 mg/kg ) pretreatment reversed LPS-induced immobility in a dose-dependent manner . DB00533 ( 2 mg/kg ) and nimesulide ( 2 mg/kg ) , P35354 inhibitors , also reversed the LPS-induced immobility , which was significantly potentiated by concomitant administration of GTE . On the other hand , GTE did not show any potentiating effect on immobility with naproxen ( 10 mg/kg ) , which is a nonselective P36551 blocker . Interestingly the antioxidant , carvedilol ( 2 mg/kg ) did not produce any effect on immobility either in normal or in LPS treated mice . The results of the study implicate the role of P35354 inhibition by GTE in the reversal of LPS-induced immobility .

MH_train_1180

MH_train_1180

MH_train_1180

interacts_with DB00175?

multiple_choice

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 . DB00175 , a P04035 inhibitor , blocks the cell cycle progression but not Ca2+ influx induced by P05019 in FRTL-5 cells . P05019 , when added to the DB00024 -primed FRTL-5 cells , induces a long lasting Ca2+ influx , and then , DNA synthesis . Moreover , Ca2+ channel agonist , B AY K8644 can mimic these effects on cell proliferation . We studied the effect of P04035 inhibitor , DB00175 on P05019 -induced cell cycle progression in FRTL-5 cells . DB00175 inhibited DNA synthesis induced both by P05019 and by BAY K8644 . In contrast , Ca2+ influx stimulated by P05019 was unaffected . These data demonstrate that the signal transduction pathway evoked by P05019 may possibly involve pravastatin-sensitive process at the downstream step of Ca2+ entry . P04035 inhibitors are known to modulate some cellular signal transduction systems by blocking the membrane attachment of low molecular weight GTP binding proteins such as P01112 . Therefore , pravastatin-sensitive process that we have shown here might possibly involve some of such small G protein . The P04035 inhibitor pravastatin stimulates insulin secretion through organic anion transporter polypeptides . The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor pravastatin has been reported to have a beneficial effect on reducing the new onset of diabetes as well as lowering plasma lipids . Because pravastatin is a water-soluble organic anion , it can not easily penetrate the lipid bilayer of the cell membrane . As the precise mechanisms of the effect of pravastatin on glucose metabolism and diabetes have not been clarified , we examined the roles of the organic anion transporter family on pravastatin-treated islet and adipocyte functions . Rat oatp1/slco1a1 , oatp2/slco1a4 and oatp3/slco1a5 were expressed in the pancreas , and rat oatp3/slco1a5 was also detected in rat insulinoma cell line P01308 -1e . DB00175 was transported not only by oatp1/slco1a1 and oatp2/slco1a4 , but also by rat oatp3/slco1a5 . DB00175 uptake into P01308 -1e cells was detected and this transport was inhibited by sulfobromophthalein and rifampicin , both of which are known to inhibit oatp family-mediated uptake . In addition , pravastatin enhanced the glucose-stimulated insulin secretion from P01308 -1e cells . When fat-loaded db/db mice were treated with pravastatin , glucose intolerance and insulin resistance were prevented . In addition , insulin secretion from isolated islets was enhanced by pravastatin . These data suggest that pravastatin has pleiotropic effects on islets through membrane transport under high fat/glucose conditions . Inflammatory signaling compromises cell responses to interferon alpha . DB05258 ( IFNα ) is widely used for treatment of melanoma and certain other malignancies . This cytokine as well as the related IFNβ exerts potent anti-tumorigenic effects ; however , their efficacy in patients is often suboptimal . Here , we report that inflammatory signaling impedes the effects of IFNα/β . Melanoma cells can secrete pro-inflammatory cytokines that inhibit cellular responses to IFNα/β via activating the ligand-independent pathway for the phosphorylation and subsequent ubiquitination and accelerated degradation of the P17181 chain of type I IFN receptor . Catalytic activity of the p38 protein kinase was required for P17181 downregulation and inhibition of IFNα/β signaling induced by proinflammatory cytokines such as interleukin 1 ( IL-1 ) . Activation of p38 kinase inversely correlated with protein levels of P17181 in clinical melanoma specimens . Inhibition of p38 kinase augmented the inhibitory effects of IFNα/β on cell viability and growth in vitro and in vivo . The roles of inflammation and p38 protein kinase in regulating cellular responses to IFNα/β in normal and tumor cells are discussed . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used . Intracellular targets of cyclin-dependent kinase inhibitors : identification by affinity chromatography using immobilised inhibitors . BACKGROUND : Chemical inhibitors of cyclin-dependent kinases ( CDKs ) have great therapeutic potential against various proliferative and neurodegenerative disorders . DB02116 , a 2,6,9-trisubstituted purine , has been optimized for activity against P06493 /cyclin B by combinatorial and medicinal chemistry efforts to yield the purvalanol inhibitors . Although many studies support the action of purvalanols against CDKs , the actual intracellular targets of 2,6 , 9-trisubstituted purines remain unverified . RESULTS : To address this issue , purvalanol B ( 95. ) and an N6-methylated , CDK-inactive derivative ( 95M. ) were immobilized on an agarose matrix . Extracts from a diverse collection of cell types and organisms were screened for proteins binding purvalanol B . In addition to validating CDKs as intracellular targets , a variety of unexpected protein kinases were recovered from the 95. matrix . Casein kinase 1 ( CK1 ) was identified as a principal 95. matrix binding protein in Plasmodium falciparum , Leishmania mexicana , Toxoplasma gondii and Trypanosoma cruzi . DB02733 compounds also inhibit the proliferation of these parasites , suggesting that CK1 is a valuable target for further screening with 2,6,9-trisubstituted purine libraries . CONCLUSIONS : That a simple batchwise affinity chromatography approach using two purine derivatives facilitated isolation of a small set of highly purified kinases suggests that this could be a general method for identifying intracellular targets relevant to a particular class of ligands . This method allows a close correlation to be established between the pattern of proteins bound to a small family of related compounds and the pattern of cellular responses to these compounds . Hepatitis C virus E2 protein promotes human hepatoma cell proliferation through the MAPK/ P29323 signaling pathway via cellular receptors . Dysregulation of mitogen-activated protein kinase ( MAPK ) signaling pathways by various viruses has been shown to be responsible for viral pathogenicity . The molecular mechanism by which hepatitis C virus ( HCV ) infection caused human liver diseases has been investigated on the basis of abnormal intracellular signal events . Current data are very limited involved in transmembrane signal transduction triggered by HCV E2 protein . Here we explored regulation of the MAPK/extracellular signal-regulated kinase ( MAPK/ P29323 ) signaling pathway by E2 expressed in Chinese hamster oval cells . In human hepatoma Huh-7 cells , E2 specifically activated the MAPK/ P29323 pathway including downstream transcription factor P39905 -2 and greatly promoted cell proliferation . P60033 and low density lipoprotein receptor ( P01130 ) on the cell surface mediated binding of E2 to Huh-7 cells . The MAPK/ P29323 activation and cell proliferation driven by E2 were suppressed by blockage of P60033 as well as P01130 . Furthermore , pretreatment with an upstream kinase Q02750 /2 inhibitor U0126 also impaired the MAPK/ P29323 activation and cell proliferation induced by E2 . Our results suggest that the MAPK/ P29323 signaling pathway triggered by HCV E2 via its receptors maintains survival and growth of target cells . 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 . Agents with selective estrogen receptor ( ER ) modulator activity induce apoptosis in vitro and in vivo in ER-negative glioma cells . Tamoxifen , a member of the selective estrogen receptor modulator ( SERM ) family , is widely used in the treatment of estrogen receptor ( ER ) -expressing breast cancer . It has previously been shown that high-dose tamoxifen has cytotoxic activity against glioma cells , but whether this effect is drug specific or represents a general property of SERMs is unknown . In this study , we demonstrate that tamoxifen and DB05487 , a novel benzopyranone with SERM activity , induce glioma cell apoptosis in a dose- and time-dependent manner . Moreover , administration of tamoxifen and DB05487 suppresses tumor growth in vivo and extends animal survival in glioma xenograft models . None of the eight glioma cell lines examined express either P03372 or -beta , suggesting the mechanism for tamoxifen- and DB05487 -induced glioma cell apoptosis is independent of the ER signaling pathway . Complementary DNA microarray expression profiling allowed us to identify a subset of genes specifically regulated by tamoxifen and DB05487 , and not by other apoptotic stimuli , including nuclear factor ( NF ) -kappaB with its target genes IEX-3 , P04179 , P05231 , and P10145 . We demonstrate that suppression of NF-kappaB activation markedly enhances SERM-induced apoptosis , suggesting a role for NF-kappaB in protecting glioma cells from SERM-induced cytotoxicity . These findings demonstrate for the first time that a SERM other than tamoxifen can induce glioma cell apoptosis in vitro and in vivo and that the clinical efficacy of SERMs for the treatment of malignant gliomas could potentially be enhanced by simultaneous inhibition of the NF-kappaB pathway . Renal changes induced by a cyclooxygenase-2 inhibitor during normal and low sodium intake . P35354 ( P35354 ) has been identified in renal tissues under normal conditions , with its expression enhanced during sodium restriction . To evaluate the role of P35354 -derived metabolites in the regulation of renal function , we infused a selective inhibitor ( nimesulide ) in anesthetized dogs with normal or low sodium intake . The renal effects elicited by nimesulide and a non-isozyme-specific inhibitor ( meclofenamate ) were compared during normal sodium intake . In ex vivo assays , meclofenamate , but not nimesulide , prevented the platelet aggregation elicited by arachidonic acid . During normal sodium intake , nimesulide infusion ( n=6 ) had no effects on arterial pressure or renal hemodynamics but did reduce urinary sodium excretion , urine flow rate , and fractional lithium excretion . In contrast , nimesulide administration increased arterial pressure and decreased renal blood flow , urine flow rate , and fractional lithium excretion during low sodium intake ( n=6 ) . P35354 inhibition reduced urinary prostaglandin E(2) excretion in both groups but did not modify plasma renin activity in dogs with low ( 8.1+/-1.1 ng angiotensin I. mL(-1). h(-1) ) or normal ( 1.8+/-0.4 ng angiotensin I. mL(-1). h(-1) ) sodium intake . DB00939 infusion in dogs with normal sodium intake ( n=8 ) induced a greater renal hemodynamic effect than nimesulide infusion . These results suggest that P35354 -derived metabolites ( 1 ) are involved in the regulation of sodium excretion in dogs with normal sodium intake , ( 2 ) play an important role in the regulation of renal hemodynamic and excretory function in dogs with low sodium intake , and ( 3 ) are not involved in the maintenance of the high renin levels during a long-term decrease in sodium intake . DB00175 improves renal ischemia-reperfusion injury by inhibiting the mevalonate pathway . Statins are known to lessen the severity of renal ischemia-reperfusion injury . The present study was undertaken to define the mechanism of renoprotective actions of statins using a mouse kidney injury model . Treatment of mice with pravastatin , a widely used statin , improved renal function after renal ischemia-reperfusion without lowering the plasma cholesterol level . Administration of pravastatin with mevalonate , a product of P04035 , eliminated renal protection suggesting an effect of pravastatin on mevalonate or its metabolism . In hypercholestrolemic apolipoprotein E knockout mice with reduced P04035 activity ; the degree of injury was less severe than in control mice , however , there was no protective action of pravastatin on renal injury in the knockout mice . Treatment with a farnesyltransferase inhibitor ( L-744832 ) mimicked pravastatin 's protective effect but co-administration with the statin provided no additional protection . Both pravastatin and L-744832 inhibited the injury-induced increase in plasma P05231 concentration to a similar extent . Our results suggest the protective effect of pravastatin on renal ischemia-reperfusion injury is mediated by inhibition of the mevalonate-isoprenoid pathway independent of its lipid lowering action . Growth inhibitory effects of IFN-beta on human liver cancer cells in vitro and in vivo . We investigated the effects of interferon-beta ( IFN-beta ) on the growth of human liver cancer cells . The effects of IFN-beta with or without 5-fluorouracil ( DB00544 ) on the proliferation of 13 liver cancer cell lines were investigated in vitro . Chronologic change in IFN-alpha receptor 2 ( P17181 -2 ) expression was monitored in hepatocellular carcinoma ( HCC ) cells ( Q86TB3 -1B ) cultured with IFN-beta . After Q86TB3 -1B cells were transplanted into nude mice , various doses of IFN-beta were administered , and the tumor volume , weight , histology , tumor blood vessel , and angiogenesis factor expression were examined . IFN-beta inhibited the growth of 11 cell lines with apoptosis in a dose-dependent and time-dependent manner . With IFN-beta , P17181 -2 expression in Q86TB3 -1B cells was significantly downregulated from 6 to 12 h . IFN-beta induced a dose-dependent decrease in tumor volume and weight and a significant increase of apoptosis in the tumor . Both basic fibroblast growth factor ( P09038 ) and blood vessel number in the tumor decreased only in mice receiving the lowest dose ( 1000 IU ) of IFN-beta . IFN-beta with 10 muM of DB00544 frequently induced synergistic antiproliferative effects . IFN-beta with or without DB00544 induces strong antitumor effects in HCC cells , and we conclude that IFN-beta is useful for the prevention and treatment of HCC . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . 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 . 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 . 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 . Inhibition of tumor cell growth , invasion , and metastasis by EXEL-2880 ( DB05030 , GSK1363089 ) , a novel inhibitor of P14210 and P15692 receptor tyrosine kinases . The DB00134 receptor tyrosine kinase and its ligand , hepatocyte growth factor ( P14210 ) , are overexpressed and/or activated in a wide variety of human malignancies . Vascular endothelial growth factor ( P15692 ) receptors are expressed on the surface of vascular endothelial cells and cooperate with DB00134 to induce tumor invasion and vascularization . EXEL-2880 ( DB05030 , GSK1363089 ) is a small-molecule kinase inhibitor that targets members of the P14210 and P15692 receptor tyrosine kinase families , with additional inhibitory activity toward P10721 , Flt-3 , platelet-derived growth factor receptor beta , and Tie-2 . Binding of EXEL-2880 to DB00134 and P15692 receptor 2 ( P35968 ) is characterized by a very slow off-rate , consistent with X-ray crystallographic data showing that the inhibitor is deeply bound in the DB00134 kinase active site cleft . EXEL-2880 inhibits cellular P14210 -induced DB00134 phosphorylation and P15692 -induced extracellular signal-regulated kinase phosphorylation and prevents both P14210 -induced responses of tumor cells and P14210 / P15692 -induced responses of endothelial cells . In addition , EXEL-2880 prevents anchorage-independent proliferation of tumor cells under both normoxic and hypoxic conditions . In vivo , these effects produce significant dose-dependent inhibition of tumor burden in an experimental model of lung metastasis . Collectively , these data indicate that EXEL-2880 may prevent tumor growth through a direct effect on tumor cell proliferation and by inhibition of invasion and angiogenesis mediated by P14210 and P15692 receptors . The glial cell modulator and phosphodiesterase inhibitor , DB05066 ( ibudilast ) , attenuates prime- and stress-induced methamphetamine relapse . Stress and renewed contact with drug ( a " slip " ) have been linked to persisting relapse of methamphetamine abuse . Human brain microglial activation has been linked with methamphetamine abuse , and inhibitors of glial cell activation , certain phosphodiesterase ( PDE ) inhibitors , and glial cell derived neurotrophic factor ( P39905 ) have been reported to modulate drug abuse effects . Our objective was to determine whether the glial cell attenuator , 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine ( DB05066 , ibudilast ) , a non-selective PDE inhibitor and promoter of P39905 , could reduce stress- and methamphetamine prime-induced reinstatement of methamphetamine-seeking behavior . Male Long-Evans hooded rats were trained to lever press reinforced with 0.1 mg/kg i.v. methamphetamine infusion according to fixed-ratio 1 ( FR1 ) reinforcement schedules during daily , 2-hour experimental sessions . After performance had stabilized , lever pressing was extinguished for 12 consecutive sessions and doses of 0 ( vehicle ) , 2.5 and 7.5 mg/kg DB05066 were then administered intraperitoneally b.i.d. on the last 2 days of extinction and then once on the testday to separate groups of 12 rats . During testing , the rats were given 15 min of intermittent footshock or a 1 mg/kg i.p. methamphetamine prime followed by a 2-hour reinstatement test session . DB05066 significantly reduced response levels of footshock-induced ( 2.5 and 7.5 mg/kg ) and prime-induced ( 7.5 mg/kg ) reinstatement of extinguished methamphetamine-maintained responding . DB05066 has properties consistent with the ability to attenuate relapse precipitated by stress and methamphetamine " slips " during abstinence . These results thus reinforce interest in atypical neurobiological mechanisms which could be exploited for developing novel medications for treating drug abuse disorders . 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 . Phase I evaluation of DB05243 , an oral , potent , and selective O60674 inhibitor . This phase I study evaluated selective O60674 inhibitor DB05243 in 30 patients with myelofibrosis . The initial dose cohorts were 100 , 200 , and 300 mg orally on days 1-21 of a 28-day cycle . Central and/or peripheral neurotoxicity developed in all patients . Subsequently , patients were treated on lower doses ; neurotoxicity was again observed , leading to study termination . Peripheral neuropathy resolved in 50 % , and central neurotoxicity in all patients within months after therapy cessation . Myelosuppression was minimal . The terminal half-life of DB05243 was approximately 21 h , with steady state reached by Day 8 . International Working Group defined responses were seen in three ( 10 % ) patients . 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 .

MH_train_1181

MH_train_1181

MH_train_1181

interacts_with DB00222?

multiple_choice

17β-estradiol inhibits P14780 and Q09428 /TrpM4 expression and activation and thereby attenuates BSCB disruption/hemorrhage after spinal cord injury in male rats . Blood-spinal cord barrier ( BSCB ) disruption and progressive hemorrhage after spinal cord injury ( SCI ) lead to secondary injury and the subsequent apoptosis and/or necrosis of neuron and glia , causing permanent neurological deficits . In this study , we examined the effect of 17β-estradiol ( E2 ) on BSCB breakdown and hemorrhage as well as subsequent inflammation after SCI . After a moderate contusion injury at the 9th thoracic segment of spinal cord , E2 ( 300 μg/kg ) was administered by iv injection immediately after SCI , and the same dose of E2 was then administered 6 and 24 hours after injury . Our data show that E2 attenuated BSCB permeability and hemorrhage and reduced the infiltration of neutrophils and macorphages after SCI . Consistent with this finding , the expression of inflammatory mediators was significantly reduced by E2 . Furthermore , E2 treatment significantly inhibited the expression of sulfonylurea receptor 1 and transient receptor potential melastatin 4 after injury , which are known to mediate hemorrhage at an early stage after SCI . Moreover , the expression and activation of matrix metalloprotease-9 after injury , which is known to disrupt BSCB , and the degradation of tight junction proteins , such as zona occludens-1 and occludin , were significantly inhibited by E2 treatment . Furthermore , the protective effects of E2 on BSCB disruption and functional improvement were abolished by an estrogen receptor antagonist , ICI 182780 ( 3 mg/kg ) . Thus , our study provides evidence that the neuroprotective effect of E2 after SCI is , in part , mediated by inhibiting BSCB disruption and hemorrhage through the down-regulation of sulfonylurea receptor 1/transient receptor potential melastatin 4 and matrix metalloprotease-9 , which is dependent on estrogen receptor . Inward rectifier potassium currents as a target for atrial fibrillation therapy . Subunits of inwardly rectifying potassium channels ( Kir ) are expressed in many different tissues of the human body . Inward rectifier currents expressed in the heart are constituted by pore-forming alpha-subunits of Kir2 , Kir3 , and Kir6 subfamilies . Characteristic properties of inward rectifiers comprise small outward conductances that nevertheless are important to terminal repolarization of cardiac action potentials . There is considerable difference in the regional expression of cardiac Kir channels , and subunits are additionally regulated by specific disease conditions . Resulting changes facilitate occurrence and persistence of atrial fibrillation ( AF ) . For instance , upregulation of Kir2.1 protein and resultant current I P04264 is a hallmark of AF-related ionic remodeling . Increased I P04264 helps to stabilize atrial rotors , and current inhibition has accordingly been suggested as an antiarrhythmic approach for AF therapy . But there are caveats to I P04264 inhibition per se , and there is no specific inhibitor of Kir2 channels . Modulation of I P04264 rectification properties seems theoretically interesting for manipulation of Kir2 currents as an antiarrhythmic approach . Kir3-based muscarinic currents ( I KACh ) are functionally upregulated during AF through increased constitutive activity ( passing current in the absence of an agonist ) . Upregulated I KACh supports sustenance of the arrhythmia . There is considerable intraatrial diversity in the expression of underlying Kir3.1/Kir3.4 subunits , but atrial-specific localization makes inhibition of this current a potentially interesting antiarrhythmic target devoid of ventricular side effects . Experimental studies of specific inhibitors indicate efficacy in various disease models . The role of I KATP remodeling under AF conditions has not been extensively studied , but present evidence indicates current downregulation and modulation of Q14654 seems less promising than that of other inward rectifiers . 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 . 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 . Morphological changes in pancreatic islets of KATP channel-deficient mice : the involvement of KATP channels in the survival of insulin cells and the maintenance of islet architecture . The DB00171 -sensitive potassium channel ( KATP channel ) is an essential ion channel involved in glucose-induced insulin secretion . The KATP channel is composed of an inwardly rectifying potassium channel , Kir6.2 , and the sulfonylurea receptor ( Q09428 1 ) ; in the pancreas it is reported to be shared by all endocrine cell types . A previous study by our research group showed that Kir 6.2-knockout mice lacked KATP channel activities and failed to secrete insulin in response to glucose , but displayed normal blood glucose levels and only mild impairment in glucose tolerance at younger ages . In some aged knockout mice , however , obesity and hyperglycemia were recognizable . The present study aimed to reveal morphological changes in pancreatic islets of Kir 6.2-knockout mice throughout life . At birth , there were no significant differences in the islet cell arrangement between the knockout mice and controls . At 14 postnatal weeks glucagon cells appeared in the central parts of islets , and this image became more pronounced with aging . In animals older than 50 weeks insulin cells decreased in numbers and intensity of insulin immunoreactivity ; most islets in 70- and 80-week-old mice were predominantly composed of glucagon cells and peptide YY ( P10082 ) -containing cells . Staining of serial sections and double staining of single sections from these old mice demonstrated the frequent coexpression of glucagon and P10082 , which is a phenotype for the earliest progenitor cells of pancreatic endocrine cells . These findings suggest that the KATP channel is important for insulin cell survival and also regulates the differentiation of islet cells . 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 . P43490 / P43490 /visfatin and cancer . P43490 / P43490 /visfatin is the rate-limiting enzyme that catalyzes the first step in NAD biosynthesis from nicotinamide and regulates growth , apoptosis and angiogenesis of mammalian cells . This enzyme was originally cloned as a putative cytokine shown to enhance the B cell precursor maturation in the presence of P13232 and stem cell factor . A number of cancers have increased expression of P43490 / P43490 /visfatin , which regulates a variety of different signaling pathways such as PI3K/Akt , P27361 /2 and P40763 . FK866/APO866 and CHS828/ DB05217 are two known inhibitors of P43490 / P43490 /visfatin and have been evaluated as anticancer agents in the clinic . This review will focus on its role in carcinogenesis and cancer progression and its inhibitors as therapeutic target for cancer treatment . 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 . Functional autoradiography of neuropeptide Y Q03519 and P49146 subtypes in rat brain using agonist stimulated [35S]GTPgammaS binding . Neuropeptide Y , one of the most abundant brain peptides , has been found to modulate several important biological functions via a family of G-protein coupled receptors . To investigate the localization of functional P01303 receptor subtypes in the rat brain , we performed agonist-induced [35S]GTPgammaS autoradiography . The Q03519 /Y4/Y5 agonist DB00149 (31) , Pro(34)- P01303 increased [35S]GTPgammaS binding in several brain areas with a regional distribution consistent with that produced when labeling adjacent sections with [ 125I ] - DB00149 (31) , Pro(34)- P10082 . The Q03519 selective antagonist BIBP3226 antagonized the DB00149 (31) , Pro(34)- P01303 stimulated increase in [35S]GTPgammaS binding in all areas examined . The P28062 agonist P06681 - P01303 stimulated [35S]GTPgamma binding in numerous brain areas with a regional distribution similar to the binding observed with [ 125I ] - DB05004 . No increase in [35S]GTPgammaS binding above basal was observed in any brain area evaluated using Y4 and Y5 selective agonists . This study demonstrates abundant Q03519 and P49146 activation in the rat brain , while evidence for functional Y4 and Y5 receptors was not observed . 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 . Effects of alpha/beta- DB01524 immune regulating hormones on bone remodeling and apoptosis in osteoblasts . A large body of evidence suggests that the immune system directly impacts bone physiology . We tested whether immune regulating hormones ( P48061 ) , 17beta- DB01524 ( beta-AED ) , 7beta,17beta-androstenetriol ( beta-AET ) or the 17alpha- DB01524 ( alpha-AED ) , and 7alpha,17beta-androstenetriol ( alpha-AET ) metabolites could directly influence bone remodeling in vitro using human fetal osteoblasts ( FOB-9 ) . The impact on bone remodeling was examined by comparing the ratio of O14788 / O00300 gene expression in response to AED and AET compounds . The alpha-AED was found to significantly increase in the ratio of O14788 / O00300 gene expression and altering the morphology of O14788 stained FOB-9 cells . Cell viability was assessed using a Live/Dead assay . Again alpha-AED was unique in its ability to reduce the proportion of viable cells , and to induce mild apoptosis of FOB-9 cells . Treatment of FOB-9 cells with WY14643 , an activator of Q07869 and -gamma , also significantly elevated the percentage of dead cells . This increase was abolished by co-treatment with GW9962 , a specific inhibitor of P37231 . Analysis of P37231 mRNA by Quantitative RT-PCR and its activation by DNA binding demonstrated that alpha-AED increased P37231 activation by 19 % , while beta-AED conferred a 37 % decrease in P37231 activation . In conclusion , alpha-AED opposed beta-AED by elevating a bone resorption scenario in osteoblast cells . The increase in O14788 / O00300 is modulated by an activation of P37231 that in turn caused mild apoptosis of FOB-9 cells . Protein kinase A alterations in endocrine tumors . Various molecular and cellular alterations of the cyclic adenosine monophosphate ( DB02527 ) pathway have been observed in endocrine tumors . Since protein kinase A ( PKA ) is a central key component of the DB02527 pathway , studies of the alterations of PKA subunits in endocrine tumors reveal new aspects of the mechanisms of DB02527 pathway alterations in human diseases . So far , most alterations have been observed for the regulatory subunits , mainly P10644 and to a lower extent , P31323 . One of the best examples of such alteration today is the multiple neoplasia syndrome Carney complex ( CNC ) . The most common endocrine gland manifestations of CNC are pituitary GH-secreting adenomas , thyroid tumors , testicular tumors , and DB01285 -independent Cushing 's syndrome due to primary pigmented nodular adrenocortical disease ( PPNAD ) . Heterozygous germline inactivating mutations of the PKA regulatory subunit RIα gene ( P10644 ) are observed in about two-third of CNC patients , and also in patients with isolated PPNAD . P10644 is considered as a tumor suppressor gene . Interestingly , these mutations can also be observed as somatic alterations in sporadic endocrine tumors . More than 120 different P10644 mutations have been found today . Most of them lead to an unstable mutant mRNA , which will be degraded by nonsense mediated mRNA decay . In vitro and in vivo functional studies are in progress to understand the mechanisms of endocrine tumor development due to PKA regulatory subunits inactivation . P10644 mutations stimulate in most models PKA activity , mimicking in some way DB02527 pathway constitutive activation . Cross-talks with other signaling pathways summarized in this review have been described and might participate in endocrine tumorigenesis . Characterization of the interaction of ingenol 3-angelate with protein kinase C . DB05013 ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate ( PMA ) in WEHI-231 , Q9BPY8 -92 , and Colo-205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO- P04264 cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation . Expression and possible role of IGF-IR in the mouse gastric myenteric plexus and smooth muscles . P01308 -like growth factor-I ( P05019 ) and its receptor ( IGF-IR ) have tremendous trophic effects on the central , peripheral and enteric neurons . The loss of IGF-IR contributes to the development of diabetic gastroparesis . However , the nature and the function of the IGF-IR(+) cells in the gastric myenteric plexus remain unclear . In this study , anti- P28329 , anti-S100β or anti-c- P10721 antibodies were used to co-label IGF-IR(+) cells and neurons , glial cells or interstitial cells of Cajal ( ICCs ) , respectively . We also generated type 1 diabetic mice ( DM ) to explore the influence of impaired P05019 /IGF-IR in the myenteric neurons . Results showed that IGF-IR was expressed in the epithelium , smooth muscles and myenteric plexi of the mouse stomach . Most of the IGF-IR(+) cells in the myenteric plexi were P28329 (+) cholinergic neurons , but not enteric glial cells and there were more IGF-IR(+) neurons and fibers in the gastric antrum than in the corpus . The IGF-IR(+)/ P28329 (+) neurons and ICCs were closely juxtaposed , but distinctly distributed in the myenteric plexus , indicating a possible role for the IGF-IR(+)/ P28329 (+) neurons in the mediation of gastric motility through ICCs . Moreover , the decrease of IGF-IR and cholinergic neurons in the myenteric plexi and smooth muscles of DM mice suggested that P05019 /IGF-IR signaling might play a role in neuron survival and neurite outgrowth , as well as stem cell factor ( P21583 ) production , which is required for the development of ICCs . Our results provide insights into the effects of P05019 /IGF-IR signaling on the development of gastrointestinal motility disorders . Refined localization and yeast artificial chromosome ( YAC ) contig -- mapping of genes and DNA segments in the 7q21-q32 region . The chromosome localizations for 159 gene and DNA segments have been refined to one of five intervals in the 7q21-132 region through hybridization analysis with a panel of somatic cell hybrid lines . Seventy-two of these chromosome 7 markers are also mapped on common or overlapping yeast artificial chromosome ( YAC ) clones . In addition , the breakpoints of chromosome rearrangement contained in five of the somatic cell hybrid lines have been defined by flanking probes within YAC contigs . To provide a framework for further mapping of the 7q21-q32 region , we have established the physical order of a set of reference markers : cen-( P08123 -D7S15- P08684 - P27169 )-D7S456- ( brea kpoint contained in cell hybrid 1EF2/3/K017 ) - P08236 -D7S186-ASL- ( P08183 - P21439 - P62879 -EPO- P22303 ) -D7S238- ( proximal breakpoint in GM1059-Rag5 ) -D7S240-(CUTL1- P05121 )- ( breakp oints in 1CF2/5/K016 and 2068Rag22-2 ) -( P31323 -D7S13)- P07942 - ( breakpoint in JSR-17S ) -DLD-D7S16-MET- P09544 - P13569 -D7S8-tel . Comparison of the effects of nimesulide and 5,5-dimethyl-3-(3-fluorophenyl)-4-(4-methylsulphonyl) phenyl-2(5H)-furanone ( DFU ) on contractions of isolated pregnant human myometrium . OBJECTIVE : To compare the effects of 5,5-dimethyl-3-(3-fluorophenyl)-4-(4-methylsulphonyl) phenyl-2(5H)-furanone ( DFU ) and nimesulide , selective P35354 inhibitors , on the amplitude and frequency of DB00761 - , oxytocin- , and P49763 (2alpha)-stimulated contractions of isolated pregnant human myometrial strips . METHODS : Isolated myometrial strips were obtained from 20 pregnant women undergoing elective cesarean section . These strips were mounted in organ baths for recording of isometric tension . The effects of cumulative concentrations of nimesulide and DFU on DB00761 - , oxytocin- , and P49763 (2alpha)-stimulated myometrial contractions were measured , and values for -log(10)EC(50) and mean maximal inhibition ( E(max) ) were compared . DB04743 ( 10(-8) to 10(-4)M ) and DFU ( 10(-8) to 10(-4)M ) inhibited in a concentration-dependent manner the DB00761 - , oxytocin- , and P49763 (2alpha)-stimulated contractions of myometrial strips , with a significant effect on the amplitude ( 10(-7) to 10(-4)M ) and the frequency ( 10(-6) to 10(-4)M ) . RESULTS : The inhibitor effect of DFU was more potent than nimesulide on DB00761 - , oxytocin- , and P49763 (2alpha)-stimulated myometrial contractions , however , the inhibitor effects of nimesulide and DFU was much greater on DB00761 -stimulated contractions than on oxytocin- and P49763 (2alpha)-stimulated myometrial contractions ( P < 0.05 ) . There was no significant difference between E(max) values of nimesulide and DFU in all tissues ( P > 0.05 ) . CONCLUSION : DFU is a more potent inhibitor than nimesulide on DB00761 - , oxytocin- , and P49763 (2alpha)-stimulated contractions of pregnant human myometrium . The inhibitor effects of nimesulide and DFU were predominantly on DB00761 -stimulated contractions . Matrix proteinase inhibition by DB05387 , a multifunctional antiangiogenic compound . BACKGROUND : Matrix metalloproteinases ( MMPs ) play an important role in tissue remodeling under normal physiological and pathological conditions and are thus attractive targets for both diagnostic and therapeutic purposes . Here , we examined the effect of DB05387 , an orally bioavailable standardized extract made of cartilage that shows significant antiangiogenic and antimetastatic properties in vivo , on the activity of various members of the MMP family . MATERIALS AND METHODS : The effect of DB05387 on the activity of MMPs was assessed by fluorimetric assays and by substrate gel zymography . RESULTS : DB05387 markedly inhibits the gelatinolytic activity of P08253 and to a lesser extent those of P03956 , P09237 , P14780 and P45452 . DB05387 also inhibited the elastinolytic activities of P08253 and P14780 as well as P39900 ( metalloelastase ) , porcine pancreatic elastase ( PPE ) , and human leukocyte elastase ( P08246 ) . Western blot analysis revealed the presence within DB05387 of immunoreactive P01033 -like proteins , suggesting that these proteins may be at least partly responsible for the observed MMP inhibition . CONCLUSIONS : Taken together , these results demonstrate that DB05387 contains P01033 -like proteins that could be responsible for the specific inhibition of MMPs . Given the recent studies suggesting the presence within this compound of specific inhibitor(s) of endothelial cell proliferation , DB05387 appears as a pleotropic agent able to interfere with several biochemical steps leading to angiogenesis and to other physiopathological conditions . Since DB05387 is currently under Phase III clinical investigations , these findings are also of considerable importance for our understanding of its anticancer properties . Survey of 548 oncogenic fusion transcripts in thyroid tumors supports the importance of the already established thyroid fusions genes . Neoplasms frequently present structural chromosomal aberrations that can alter the level of expression of a protein or to the expression of an aberrant chimeric protein . In the thyroid , the Q06710 - P37231 fusion is present in the neoplastic lesions that have a follicular architecture-follicular thyroid carcinoma ( FTC ) and follicular variant of papillary thyroid carcinoma ( FVPTC ) , and less frequently in follicular thyroid adenoma ( DB00499 ) , while the presence of P07949 /PTC fusions are largely restricted to papillary thyroid carcinoma ( PTC ) . The ability to detect fusion genes is relevant for a correct diagnosis and for therapy . We have developed a new fusion gene microarray-based approach for simultaneous analysis of all known and predicted fusion gene variants . We did a comprehensive screen for 548 known and putative fusion genes in 27 samples of thyroid tumors and three positive controls-one thyroid cancer cell line ( TPC-1 ) and two PTCs with known Q16204 - P07949 ( alias P07949 / Q13635 ) fusion gene , using this microarray . Within the thyroid tumors tested , only well known , previously reported fusion genes in thyroid oncology were identified . Our results reinforce the pathogenic role played by P07949 / Q13635 , P07949 /PTC3 , and Q06710 - P37231 fusion genes in thyroid tumorigenesis . Genetics in gestational diabetes mellitus : association with incidence , severity , pregnancy outcome and response to treatment . Constant advances in gene mapping technology have allowed research to focus from rare monogenic disorders on common complex diseases involving multiple susceptibility genes-environment interactions . Gestational diabetes mellitus ( GDM ) is a heterogeneous pathogenic condition affecting 2-5 % of all pregnant women during pregnancy . GDM is considered to result when genetic predisposition is triggered by increased insulin resistance during pregnancy leading to what seems to be one of the primary characteristics of GDM , the pancreatic b-cell impairment . Genetic predisposition to GDM has been suggested given the occurrence of the disease within family members . Furthermore , GDM is reported to be often present in women with maturity onset diabetes of the young ( MODY ) gene mutations . In addition , candidate susceptibility gene variants have been suggested to increase the risk of GDM . These genes include glucokinase ( GCK ) , HLA antigens , insulin receptor ( P06213 ) , insulin-like growth factor-2 ( P01344 ) , P41235 , insulin gene ( P01308 -VNTR ) , plasminogen activator inhibitor 1 ( P05121 ) , potassium inwardly rectifying channel subfamily J , member 11 ( Q14654 ) , hepatocyte nuclear factor-4a ( P41235 ) . Identification of the possible underlying genetic factors of GDM would eventually enrich our knowledge on the pathophysiologic mechanism of the disease and contribute to the individualization of both prevention and treatment of complications for the mother and fetus . However , so far , little is known about the genetic basis of GDM and its potential clinical significance . This review focuses on possible gestational diabetes mellitus susceptibility genes and their association with the disease incidence and severity as well as the pregnancy outcome and the response to treatment . Validation of ITD mutations in P36888 as a therapeutic target in human acute myeloid leukaemia . Effective targeted cancer therapeutic development depends upon distinguishing disease-associated ' driver ' mutations , which have causative roles in malignancy pathogenesis , from ' passenger ' mutations , which are dispensable for cancer initiation and maintenance . Translational studies of clinically active targeted therapeutics can definitively discriminate driver from passenger lesions and provide valuable insights into human cancer biology . Activating internal tandem duplication ( ITD ) mutations in P36888 ( P36888 -ITD ) are detected in approximately 20 % of acute myeloid leukaemia ( AML ) patients and are associated with a poor prognosis . Abundant scientific and clinical evidence , including the lack of convincing clinical activity of early P36888 inhibitors , suggests that P36888 -ITD probably represents a passenger lesion . Here we report point mutations at three residues within the kinase domain of P36888 -ITD that confer substantial in vitro resistance to DB05213 ( quizartinib ) , an active investigational inhibitor of P36888 , P10721 , P16234 , P09619 and P07949 ; evolution of DB05213 -resistant substitutions at two of these amino acid positions was observed in eight of eight P36888 -ITD-positive AML patients with acquired resistance to DB05213 . Our findings demonstrate that P36888 -ITD can represent a driver lesion and valid therapeutic target in human AML . DB05213 -resistant P36888 kinase domain mutants represent high-value targets for future P36888 inhibitor development efforts . [ 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 . DB11582 suppresses osteoclastogenesis induced by O14788 and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 -induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 suppressed osteoclastogenesis induced by O14788 , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 -induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss . 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 .

MH_train_1182

MH_train_1182

MH_train_1182

interacts_with DB04946?

multiple_choice

Rapid modulation of spine morphology by the 5- Q13049 serotonin receptor through kalirin-7 signaling . The 5-HT(2A) serotonin receptor is the most abundant serotonin receptor subtype in the cortex and is predominantly expressed in pyramidal neurons . The 5-HT(2A) receptor is a target of several hallucinogens , antipsychotics , anxiolytics , and antidepressants , and it has been associated with several psychiatric disorders , conditions that are also associated with aberrations in dendritic spine morphogenesis . However , the role of 5-HT(2A) receptors in regulating dendritic spine morphogenesis in cortical neurons is unknown . Here we show that the 5-HT(2A) receptor is present in a subset of spines , in addition to dendritic shafts . It colocalizes with P78352 and with multiple PDZ protein-1 ( O75970 ) in a subset of dendritic spines of rat cortical pyramidal neurons . O75970 is enriched in postsynaptic density ( A5PKW4 ) fractions , is targeted to spines in pyramidal neurons , and enhances the localization of 5-HT(2A) receptors to the cell periphery . 5-HT(2A) receptor activation by the 5-HT(2) receptor agonist DOI induced a transient increase in dendritic spine size , as well as phosphorylation of P38936 -activated kinase ( PAK ) in cultured cortical neurons . PAK is a downstream target of the neuronal Rac guanine nucleotide exchange factor ( RacGEF ) kalirin-7 that is important for spine remodeling . O60229 -7 regulates dendritic spine morphogenesis in neurons but its role in neuromodulator signaling has not been investigated . We show that peptide interference that prevents the localization of kalirin-7 to the postsynaptic density disrupts DOI-induced PAK phosphorylation and spine morphogenesis . These results suggest a potential role for serotonin signaling in modulating spine morphology and kalirin-7 's function at cortical synapses . In vivo participation of nitric oxide in hyperproliferative epidermal phenomena in mice . A significant involvement of nitric oxide ( NO ) in the process of keratinocyte proliferation is reported with many divergences . To determine the involvement of NO in the hyperproliferative process of epidermis in vivo , non-selective inhibitor ( N(G)-nitro-L-arginine-methyl ester.HCl : L-NAME ) and selective inhibitors for inducible NO synthase ( P35228 ) and neuronal NO synthase ( P29475 ) ( DB02533 : AG and DB02207 : 7-NI , respectively ) and a NO-donor ( Sodium nitroprusside : SNP ) were topically applied twice a day in mice ear treated with multiple applications of croton oil . L-NAME and 7-NI treatments decreased and SNP increased ear edema formation . However , ear weight was reduced in groups that received L-NAME and 7-NI , while the AG and SNP groups presented an increment . The histological evaluation of epidermis thickness showed that all NOS inhibitors were able to prevent the increase in epidermis width caused by croton oil , while SNP contributed to enlargement . The same results were observed in the P12004 staining , where treatments with NOS inhibitors caused a reduction in the number of cells in the epidermis , while SNP caused an enhancement . 7-NI treatment reduced polymorphonuclear and mononuclear leukocytes migration when compared to the control group . The AG application increased the migration of polymorphonuclear and mononuclear cells , while the SNP enhanced only the polymorphonuclear cells . Therefore , in the skin NO produced by P29475 is involved in the control of keratinocyte hyperproliferation , with the contribution of P35228 . In the animal model of cutaneous chronic inflammation by croton oil , NO is involved in the exudation and leukocyte migration , with participation of all three enzymes . To press or not to press ? Differential receptor expression and response to novelty in rats learning an operant response for reward . Learning to perform instrumental tasks is an ability of all animals . In a population of rats , not all individuals will acquire an operant response for reward . We hypothesized that there could be a genetic explanation for differences between High Consumers ( those that acquired the lever press response ) and Low Consumers ( lever press response is low ) . Additionally , we proposed that this genetic difference could produce measurable changes in response to novelty . Wistar rats were trained to lever press for a 0.2 % saccharin reward and on the 10th day they were placed in a novel open field for 30 min to record locomotor activity . The prefrontal cortex and hippocampus were dissected and qPCR was used to measure mRNA expression . A significant difference ( p=.048 ; 2-way Q9UNW9 ) in gene expression was observed between Low and High Consumers . A principal component analysis ( DB11245 ) , to cluster which genes represent this difference , identified 4 genes ; 5- Q13049 and mGlu1 in the hippocampus and AMPA GluR1 and adrenergic alpha2A in the prefrontal cortex . Response to a novel open field also differed since Low Consumers displayed a higher Total Distance in comparison to High Consumers . Additionally , Low Consumers could be subdivided into Low-Lever ( with lever press response only when water deprived ) and Low-Non-Lever ( lever press response is low throughout training ) . DB11245 with this subdivision identified an additional nine genes differing within the divisions ; DB01221 Q13224 and GABAAalpha3 in the prefrontal cortex and adrenergic alpha2B and alpha2A , AMPA GluR1 , GluR2 and GluR3 , P28222 and GABAAalpha5 in the hippocampus . No effect of the oral neutral endopeptidase inhibitor candoxatril , on bronchomotor tone and histamine reactivity in asthma . P08473 ( NEP ) is found in many tissues in man , including the lung . Metabolism by NEP is one of the main mechanisms for the clearance of atrial natriuretic peptide ( P01160 ) , a hormone that causes bronchodilation and reduces nonspecific bronchial reactivity in man . DB00616 , an oral NEP inhibitor has been shown to elevate circulating P01160 levels . We have sought to determine whether the administration of candoxatril will alter bronchomotor tone ( forced expiratory volume in one second ( FEV1 ) ) and histamine reactivity . Ten male asthmatic patients with stable asthma were enrolled ( mean ( SD ) age 32 ( 10 ) yrs ; FEV1 92 ( 11 ) % predicted ) in a randomized , double-blind , placebo-controlled study . On each study day , after baseline spirometry , patients received 200 mg of candoxatril or placebo . Spirometry was repeated at half hourly intervals . After 2 h a histamine inhalation test was performed . There was no significant difference in FEV1 values at baseline or at 2 h post-dosing between active and placebo study days , with mean ( SEM ) FEV1 at baseline and 2 h of 3.71 ( 0.29 ) l and 3.85 ( 0.29 ) l on the placebo day , and 3.89 ( 0.27 ) l and 4.05 ( 0.82 ) l on the active day , respectively . The geometric mean ( range ) provocative concentration of histamine producing a 20 % fall in FEV1 ( PC20 ) on the placebo day and active day did not differ significantly , being 1.17 ( 0.25-25.8 ) and 0.93 ( 0.13-32 ) mg.ml-1 , respectively. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . The human plasma proteome : analysis of Chinese serum using shotgun strategy . We have investigated the serum proteome of Han-nationality Chinese by using shotgun strategy . A complete proteomics analysis was performed on two reference specimens from a total of 20 healthy donors , in which each sample was made from ten-pooled male or female serum , respectively . The methodology used encompassed ( 1 ) removal of six high-abundant proteins ; ( 2 ) tryptic digestion of low- and high-abundant proteins of serum ; ( 3 ) separation of peptide mixture by RP-HPLC followed by P19957 -MS/MS identification . A total of 944 nonredundant proteins were identified under a stringent filter condition ( X(corr) > or = 1.9 , > or = 2.2 , and > or = 3.75 , < or = C(n) > or = 0.1 , and R(sp) > or = 4.0 ) in both pooled male and female samples , in which 594 and 622 entire proteins were found , respectively . Compared with the total 3020 protein identifications confirmed by more than one laboratory or more than one specimen in HUPO Plasma Proteome Project ( PPP ) participating laboratories recently , 206 proteins were identified with at least two distinct peptides per protein and 185 proteins were considered as high-confidence identification . Moreover , some lower abundance serum proteins ( ng/mL range ) were detected , such as complement P01031 and Q8WXI7 , routinely used as an ovarian cancer marker in plasma and serum . The resulting nonredundant list of serum proteins would add significant information to the knowledge base of human plasma proteome and facilitate disease markers discovery . 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 . 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 . Selective inhibition of the tumor marker O60218 by antiinflammatory N-phenylanthranilic acids and glycyrrhetic acid . A human aldose reductase-like protein , O60218 in the aldo-keto reductase ( AKR ) superfamily , was recently identified as a tumor marker of several types of cancer . DB02383 , an aldose reductase inhibitor ( Q9Y4X5 ) , is known to be the most potent inhibitor of the enzyme . In this study , we compared the inhibitory effects of other ARIs including flavonoids on O60218 and aldose reductase to evaluate their specificity . However , ARIs showed lower inhibitory potency for O60218 than for aldose reductase . In the search for potent and selective inhibitors of O60218 from other drugs used clinically , we found that non-steroidal antiinflammatory N-phenylanthranilic acids , diclofenac and glycyrrhetic acid competitively inhibited O60218 , showing K(i) values of 0.35-2.9 microM and high selectivity to this enzyme ( 43-57 fold versus aldose reductase ) . Molecular docking studies of mefenamic acid and glycyrrhetic acid in the O60218 -nicotinamide adenine dinucleotide phosphate ( NADP(+) ) complex and site-directed mutagenesis of the putative binding residues suggest that the side chain of Val301 and a hydrogen-bonding network among residues Val301 , Gln114 and Ser304 are important for determining the inhibitory potency and selectivity of the non-steroidal antiinflammatory drugs . Thus , the potent and selective inhibition may be related to the cancer chemopreventive roles of the drugs , and their structural features may facilitate the design of new anti-cancer agents targeting O60218 . 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 . Association of the Q8IXF0 gene and five other loci with response to the antipsychotic iloperidone identified in a whole genome association study . A whole genome association study was performed in a phase 3 clinical trial conducted to evaluate a novel antipsychotic , iloperidone , administered to treat patients with schizophrenia . Genotypes of 407 patients were analyzed for 334,563 single nucleotide polymorphisms ( SNPs ) . SNPs associated with iloperidone efficacy were identified within the neuronal DB00233 domain protein 3 gene ( Q8IXF0 ) , close to a translocation breakpoint site previously observed in a family with schizophrenia . Five other loci were identified that include the XK , Kell blood group complex subunit-related family , member 4 gene ( Q5GH76 ) , the tenascin-R gene ( Q92752 ) , the glutamate receptor , inotropic , AMPA 4 gene ( P48058 ) , the glial cell line-derived neurotrophic factor receptor-alpha2 gene ( O00451 ) , and the NUDT9P1 pseudogene located in the chromosomal region of the serotonin receptor 7 gene ( P34969 ) . The study of these polymorphisms and genes may lead to a better understanding of the etiology of schizophrenia and of its treatment . These results provide new insight into response to iloperidone , developed with the ultimate goal of directing therapy to patients with the highest benefit-to-risk ratio . The pancreas-brain axis : insight into disrupted mechanisms associating type 2 diabetes and Alzheimer 's disease . Epidemiological and observational studies indicate a positive correlation between type 2 diabetes ( T2DM ) and dementia , with an increased risk of dementia and Alzheimer 's disease ( AD ) associated with insulin-treated diabetes patients . The purpose of this review is to reveal the molecular mechanisms that connect physiological and pathological processes commonly observed in T2DM and AD . Conformational modifications in peptide residues , such as amyloid-β peptide in AD and amylin in T2DM have been shown to instigate formation of insoluble protein aggregates that get deposited in extracellular spaces of brain and pancreatic tissue thus disrupting their normal function . Impaired insulin signaling plays a critical role in AD pathogenesis by reducing P41252 -associated P19957 kinase activity and increasing GSK-3β activity . GSK-3β has been suggested to be a component of the γ-secretase complex and is involved in amyloid-β protein precursor processing . GSK-3β along with Q00535 is responsible for hyperphosphorylation of tau leading to the formation of neurofibrillary tangles . In summary , there is evidence to believe that a molecular link connects AD and T2DM and has potential for further investigation toward development of an effective therapeutic target . Acute ethanol preexposure promotes liver regeneration after partial hepatectomy in mice by activating P05091 . It is known that chronic ethanol significantly impairs liver regeneration . However , the effect of acute ethanol exposure on liver regeneration remains largely unknown . To address this question , C57Bl6/J mice were exposed to acute ethanol ( 6 g/kg intragastrically ) for 3 days , and partial hepatectomy ( PHx ) was performed 24 h after the last dose . Surprisingly , acute ethanol preexposure promoted liver regeneration . This effect of ethanol did not correlate with changes in expression of cell cycle regulatory genes ( e.g. , cyclin D1 , P38936 , and p27 ) but did correlate with protection against the effect of PHx on indices of impaired lipid and carbohydrate metabolism . DB00898 preexposure protected against inhibition of the oxidant-sensitive mitochondrial enzyme , aconitase . The activity of aldehyde dehydrogenase 2 ( P05091 ) was significantly increased by ethanol preexposure . The effect of ethanol was blocked by inhibiting ( DB02115 ) and was mimicked by activating ( Alda-1 ) P05091 . Lipid peroxides are also substrates for P05091 ; indeed , alcohol preexposure blunted the increase in lipid peroxidation ( 4OH-nonenal adducts ) caused by PHx . Taken together , these data suggest that acute preoperative ethanol exposure " preconditions " the liver to respond more rapidly to regenerate after PHx by activating mitochondrial P05091 , which prevents oxidative stress in this compartment . Discrimination between agonists and antagonists by the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid-selective glutamate receptor . A mutation analysis of the ligand-binding domain of P48058 subunit . The crystal structures of the ligand-binding core of the agonist complexes of the glutamate receptor-B ( P42262 ) subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid ( AMPA ) -selective glutamate receptor indicate that the distal anionic group of agonist molecules are stabilized by interactions with an N-terminal region of an alpha-helix ( helix F ) in the lobe 2 ( " domain 2 , " Armstrong , N. , and Gouaux , E. ( 2000 ) Neuron 28 , 165-181 ) of the two-lobed ligand-binding domain . We used site-directed mutagenesis to further analyze the role of this region in the recognition of both agonists and antagonists by the AMPA receptor . Wild-type and mutated versions of the ligand-binding domain of P48058 were expressed in insect cells as secreted soluble polypeptides and subjected to binding assays using [(3)H]AMPA , an agonist , and [(3)H]Ro 48-8587 ( 9-imidazol-1-yl-8-nitro-2,3,5,6-tetrahydro[1,2,4]triazolo[1,5-c] quinazoline-2,5-dione ) , a high affinity AMPA receptor antagonist , as radioligands . Single alanine substitutions at residues DB00149 -672 and DB00156 -677 severely affected the affinities for all agonists , as seen in ligand competition assays , whereas similar mutations at residues DB00128 -673 , DB00133 -674 , DB00145 -675 , DB00133 -676 , and Lys-678 selectively affected the binding affinities of one or two of the agonists . In striking contrast , the binding affinities of [(3)H]Ro 48-8587 and of another competitive antagonist , DB03759 , were not affected by any of these alanine mutations , suggesting the absence of critical side-chain interactions . Together with ligand docking experiments , our results indicate a selective engagement of the side chains of the helix F region in agonist binding , and suggest that conformational changes involving this region may play a critical role in receptor activation . Method for isolating tight-binding inhibitors of rat lens aldose reductase . Numerous animal studies indicate that aldose reductase inhibitors ( ARIs ) are beneficial for the prevention or amelioration of diabetic complications such as neuropathy , nephropathy and the ocular complications of cataract , retinopathy and keratopathy . To aid in the identification of novel potent ARIs , we have previously developed a screening method that is based on the formation of a non-covalent ternary tight-binding enzyme-inhibitor-nucleotide ( AR- Q9Y4X5 -NADPH ) complex that can be isolated using YM-10 filter units . Here , we report a modification of this method that permits us to rapidly identify tight binding ARIs that are isolated by denaturation from AR- Q9Y4X5 -NADPH complexes that are free of possible contamination resulting from the reaction of methanol with the YM-10 filter units . For the development of this procedure , nine structurally diverse ARIs were mixed with purified recombinant rat lens aldose reductase ( RLAR ) bound with NADPH to form tight-binding RLAR- Q9Y4X5 -NADPH complexes . These complexes were purified by high pressure Sephadex 75 size exclusion chromatography using ammonium acetate buffer and the formation of each complex was confirmed by electrospray ionisation mass spectrometry ( P19957 -MS ) . Each of the complexes was then denatured with methanol , rechromatographed on the size exclusion column , and the identity of the bound ARIs was confirmed by P19957 -MS . The apparent Q9Y4X5 binding with aldose reductase to form a tight binding Q9Y4X5 complex appeared proportional to their IC50 values . This procedure allows for the rapid identification of tight binding ARIs with apparent IC50s < 0.1 microm . Binding of antipsychotic drugs to human brain receptors focus on newer generation compounds . Using radioligand binding assays and post-mortem normal human brain tissue , we obtained equilibrium dissociation constants ( K(d)s ) for nine new antipsychotic drugs ( iloperidone , melperone , olanzapine , ORG 5222 , quetiapine , risperidone , sertindole , ziprasidone , and zotepine ) , one metabolite of a new drug ( 9-OH-risperidone ) , and three older antipsychotics ( clozapine , haloperidol , and pimozide ) at nine different receptors ( alpha1-adrenergic , alpha2-adrenergic , dopamine D2 , histamine H1 , muscarinic , and serotonin P08908 , P28221 , 5- Q13049 , and P28335 receptors ) . DB04946 was the most potent drug at the two adrenergic receptors . ORG 5222 was the most potent drug at dopamine D2 and 5-HT2c receptors , while ziprasidone was the most potent compound at three serotonergic receptors ( P08908 , P28221 , and 5- Q13049 ) . At the remaining two receptors , olanzapine was the most potent drug at the histamine H1 receptor ( Kd=0.087 nM ) ; clozapine at the muscarinic receptor ( Kd=9 nM ) . Certain therapeutic and adverse effects , as well as certain drug interactions can be predicted from a drug 's potency for blocking a specific receptor . These data can provide guidelines for the clinician in the choice of antipsychotic drug . Dexamethasone reverses adrenalectomy-induced neuronal de-differentiation in midbrain raphe-hippocampus axis . Differentiation leads to specific morphological and biochemical characteristics . We examined whether epigenetic factors ( e.g. , glucocorticoids ) are required to maintain neuronal differentiation in the adult brain . In the midbrain , adrenalectomy ( P10109 ) ( 1-2 wk ) reduced the size of tryptophan hydroxylase ( WH ) -immunoreactive ( IR ) neurons . P10109 rats exposed to short-term ( 24-72-h ) dexamethasone ( ST-DEX ) in the drinking saline ( 10 mg/l ) showed an increase in WH protein , somal area and dendritic size of WH-IR neurons . In the hippocampus , P10109 for 2-3 mo ( long-term ; LT ) reduced Nissl staining , calbindin ( DB09061 ) -IR and P08908 receptor mRNA in the granular cell layer , and the size of the molecular layer and its DB09061 -IR dendrites . Small vimentin ( Vim ) -IR glial cells emerged in the granular layer . ST-DEX after LT- P10109 rapidly induced a recovery of P08908 mRNA , Nissl labeling and DB09061 -IR in the granule cell layer . In the molecular layer , there was an increase in the area and in the number of DB09061 -IR dendrites . Furthermore , the Vim-IR glial cells were enlarged in size and branching . The rate of cell proliferation was studied in these animals . Immunostaining with antibodies against proliferating cell nuclear antigen ( P12004 ) and use of bromouridine argue against enhanced neurogenesis after ST-DEX in LT- P10109 . We propose that glucocorticoids induce and maintain differentiation of serotonergic and DB09061 -IR neurons in the midbrain-hippocampal axis . A neuronotrophic role for the glial P08908 receptor is suggested . Decreased social behaviour following 3,4-methylenedioxymethamphetamine ( DB01454 ) is accompanied by changes in 5- Q13049 receptor responsivity . This study examined the involvement of the 5-HT(2A) receptor in the long-term anxiogenic effect of a brief exposure of young rats to 3,4-methylenedioxymethamphetamine ( DB01454 ) using the social interaction and elevated plus-maze paradigms . Wistar rats ( post-natal day ( P01160 ) 28 ) received either DB01454 ( 5 mg/kg i.p. ) or saline ( 1 ml/kg i.p. ) hourly for 4 h on 2 consecutive days . Locomotor activity was measured for 60 min after the first injection and core body temperature was recorded at regular intervals over 4 h . On P01160 84 , without further drug administration , social interaction was assessed between treatment-matched rat pairs derived from separate litters . On P01160 86 , rats received either the 5-HT(2A/2C) receptor agonist , 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ( DOI , 1 mg/kg i.p. ) or saline and locomotor activity , wet-dog shakes and back muscle contractions were monitored . The change in elevated plus-maze behaviour was assessed following the same injection on P01160 87 . Acutely , DB01454 produced a significant hyperlocomotion and hyperthermia ( p < 0.01 ) . Following 55 days of abstinence , social interaction was reduced by 27 % in DB01454 pre-treated rats compared with that in controls ( p < 0.01 ) . On the elevated plus-maze , pre-treatment with DB01454 prevented the anxiogenic effect of DOI . On P01160 92 , hippocampal , frontal cortical and striatal 5-hydroxytryptamine ( 5-HT ) was significantly reduced in DB01454 pre-treated rats by between 16 % and 22 % , without any accompanying change in [(3)H]paroxetine binding in cortical homogenates . In conclusion , exposure of young rats to repeated DB01454 caused serotonin depletion and induced ' anxiety-like ' behaviour in the social interaction test accompanied by a long-lasting reduction in specific 5-HT(2A) receptor mediated behaviour . Complementary roles for amygdala and periaqueductal gray in temporal-difference fear learning . Pavlovian fear conditioning is not a unitary process . At the neurobiological level multiple brain regions and neurotransmitters contribute to fear learning . At the behavioral level many variables contribute to fear learning including the physical salience of the events being learned about , the direction and magnitude of predictive error , and the rate at which these are learned about . These experiments used a serial compound conditioning design to determine the roles of basolateral amygdala ( BLA ) DB01221 receptors and ventrolateral midbrain periaqueductal gray ( vlPAG ) mu-opioid receptors ( MOR ) in predictive fear learning . Rats received a three-stage design , which arranged for both positive and negative prediction errors producing bidirectional changes in fear learning within the same subjects during the test stage . Intra-BLA infusion of the Q13224 receptor antagonist DB08954 prevented all learning . In contrast , intra-vlPAG infusion of the MOR antagonist CTAP enhanced learning in response to positive predictive error but impaired learning in response to negative predictive error -- a pattern similar to Hebbian learning and an indication that fear learning had been divorced from predictive error . These findings identify complementary but dissociable roles for amygdala DB01221 receptors and vlPAG MOR in temporal-difference predictive fear learning . Induction of Q8WXI7 -specific B and T cell responses in patients injected with MAb- DB04964 -- evidence for antibody-mediated antigen-processing and presentation of Q8WXI7 in vivo . The murine monoclonal anti- Q8WXI7 antibody MAb- DB04964 has previously been administered as an immunoscintigraphic agent in order to monitor recurrence of ovarian cancer in patients , and a long-term follow-up demonstrated a survival benefit for these patients . The clinical benefit was initially attributed to the activation of the idiotypic network . The objective of this study was to investigate the role of Q8WXI7 -MAb- DB04964 immune complex formation on the induction of Q8WXI7 -specific immune responses . Analysis of patient serum samples from pharmacokinetic studies demonstrated that the antibody forms immune complexes with Q8WXI7 in circulation within 30 minutes of injection . Induction of humoral and cellular anti- Q8WXI7 responses correlated with the amount of circulating Q8WXI7 antigen present at time of antibody injection . Subsequent to the injection of MAb- DB04964 , the patients generated anti- Q8WXI7 antibodies that were directed against various epitopes on the antigen and were not restricted to the specific epitope recognized by MAb- DB04964 . The generation of Q8WXI7 -specific B and T cell responses after MAb- DB04964 injection correlated with improved survival . The influence of circulating Q8WXI7 for the induction of Q8WXI7 -specific immune responses and the multi-epitopic nature of the human anti- Q8WXI7 antibodies suggest that the majority of these antibodies were not induced via the idiotypic network but by the autologous antigen itself . Since antibody and T cell responses to Q8WXI7 were not present before injection of MAb- DB04964 , it is hypothesized that complex formation of MAb- DB04964 with circulating antigen triggers the induction of Q8WXI7 -specific immune responses . Inhibition of cyclin-dependent kinases , GSK-3beta and CK1 by hymenialdisine , a marine sponge constituent . BACKGROUND : Over 2000 protein kinases regulate cellular functions . Screening for inhibitors of some of these kinases has already yielded some potent and selective compounds with promising potential for the treatment of human diseases . RESULTS : The marine sponge constituent hymenialdisine is a potent inhibitor of cyclin-dependent kinases , glycogen synthase kinase-3beta and casein kinase 1 . DB02950 competes with DB00171 for binding to these kinases . A P24941 -hymenialdisine complex crystal structure shows that three hydrogen bonds link hymenialdisine to the Glu81 and Leu83 residues of P24941 , as observed with other inhibitors . DB02950 inhibits Q00535 /p35 in vivo as demonstrated by the lack of phosphorylation/down-regulation of Pak1 kinase in E18 rat cortical neurons , and also inhibits GSK-3 in vivo as shown by the inhibition of P46821 phosphorylation . DB02950 also blocks the in vivo phosphorylation of the microtubule-binding protein tau at sites that are hyperphosphorylated by GSK-3 and Q00535 /p35 in Alzheimer 's disease ( cross-reacting with Alzheimer's-specific AT100 antibodies ) . CONCLUSIONS : The natural product hymenialdisine is a new kinase inhibitor with promising potential applications for treating neurodegenerative disorders .

MH_train_1183

MH_train_1183

MH_train_1183

interacts_with DB06822?

multiple_choice

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 . Involvement of gene polymorphisms of the folate pathway enzymes in gene expression and anticancer drug sensitivity using the NCI-60 panel as a model . DB00158 , a vitamin of the B group involved in one-carbon group metabolism , plays an important role in DNA synthesis and methylation . Several polymorphisms in the genes involved in folate uptake and biotransformations have been shown to be associated to the risk of cancer and to anticancer drug response . We studied common polymorphisms in P42898 ( N(5,10)-methylene- DB00116 reductase ) , P11586 ( N(5,10)-methylene- DB00116 dehydrogenase ) , Q99707 ( methionine synthetase ) and P41440 ( reduced folate carrier ) in the panel of 60 human tumour cell lines established by the NCI for anticancer drug screening and we tentatively associated these polymorphisms with gene expression and drug cytotoxicity as extracted from the public database of the Developmental Therapeutic Programme . We observed a consistent and highly significant association between the presence of the variant C allele of the A > C1298 polymorphism of P42898 and the sensitivity to many anticancer drugs belonging to the classes of antifolates , antimetabolites , alkylating agents and , to a lesser extent , topoisomerase inhibitors . In contrast , the T variant allele of the C > T677 variation of P42898 was rather associated to lower sensitivity of the cell lines towards anticancer drugs ( alkylating agents , antifolates and antimetabolites ) but with much lower effects than the A > C1298 variation . The polymorphisms of the other genes studied were not associated with differences in anticancer drug sensitivity , but the expression of the P41440 gene was significantly correlated with the sensitivity to several drugs ( antifolates , thiopurines , nitrosoureas , and Q9UI36 -platinum drugs ) . We concluded that the NCI-60 panel may constitute a good starting point for implementing clinical studies aimed at discovering and validating predictive genetic markers of drug efficacy and/or toxicity . P05112 synthesis by in vivo-primed memory P01730 + T cells : II . Presence of P05112 is not required for P05112 synthesis in primed P01730 + T cells . Previous studies have shown that the presence of P05112 is required for the development of P05112 synthesis in naive P01730 + T cells . The purpose of our current studies was to investigate the role of P05112 in the development of P05112 synthesis in primed memory T cells . We therefore examined P01730 + T cells taken from lymph nodes of BALB/c mice immunized with DB05299 ( KLH ) and restimulated in vitro with KLH . Our results with such primed resting P01730 + T cells programmed to produce P05112 indicated that the production of P05112 did not require the presence of P05112 ( although the presence of P60568 was absolutely necessary ) , and was only slightly limited by the presence of anti- P05112 MAb . These results with resting memory T cells were not biased by the presence of activated T cells already producing substantial quantities of P05112 , since we demonstrated that high-density memory T cells could produce P05112 in the absence of P05112 , and because T cells that actively produce P05112 do not persist in vivo very long after antigen exposure . These results indicate that P05112 synthesis in T cells committed to P05112 production can indeed occur in the absence of P05112 when culture conditions have been optimized and suggest that therapies with anti- P05112 MAb or with soluble P05112 receptors designed to control the development of P05112 synthesis in memory T cells from individuals exhibiting excessive P05112 synthesis will be unsuccessful . Therefore , other therapies , for example , utilizing IL-12 , will be required to modulate the relatively fixed programs in memory T cells that direct the development of cytokine synthesis . Thalidomide suppresses Up-regulation of human immunodeficiency virus coreceptors P61073 and P51681 on P01730 + T cells in humans . Concurrent infection in patients with human immunodeficiency virus ( HIV ) infection increases the expression of HIV coreceptors P61073 and P51681 . Thalidomide has beneficial effects in a number of HIV-associated diseases . The effect of thalidomide on P61073 and P51681 expression on P01730 + T cells was determined . Thalidomide produced a dose-dependent inhibition of lipopolysaccharide ( LPS ) -induced up-regulation of P61073 and P51681 in vitro . Antibody to tumor necrosis factor-alpha ( P01375 ) also attenuated the LPS-induced HIV coreceptor up-regulation , which was not further reduced by thalidomide . Thalidomide ( 400 mg ) was orally administered to 6 men , and their blood was stimulated ex vivo with LPS , staphylococcal or mycobacterial antigens , or antibody to CD3 or P10747 cells . All stimuli induced up-regulation of HIV coreceptors , which was reduced after ingestion of thalidomide . Thalidomide may be beneficial in the treatment of intercurrent infections during HIV infection by reducing the up-regulation of P61073 and P51681 expression on P01730 + T cells induced by bacterial and mycobacterial antigens , by a mechanism that involves inhibition of P01375 . 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 . DB00114 ( PLP ) deficiency might contribute to the onset of type I diabetes . The incidence of type I diabetes is rising worldwide , particularly in young children . Type I diabetes is considered a multifactorial disease with genetic predisposition and environmental factors participating . Currently , despite years of research , there is no consensus regarding the factors that initiate the autoimmune response . Type I diabetes is preceded by autoimmunity to islet antigens , among them the protein glutamic acid decarboxylase , Q05329 . DB00114 ( PLP ) is formed from vitamin B6 by the action of pyridoxal kinase . Interaction of Q05329 with PLP is necessary for Q05329 -mediated synthesis of the neurotransmitter γ-aminobutyric acid ( GABA ) . PLP is also a required cofactor for dopamine synthesis by L-aromatic decarboxylase ( L- P20711 ) . Both Q05329 and L- P20711 are expressed in pancreatic islets . Here it is proposed that lack of the vitamin B6 derivative pyridoxal 5'-phosphate might contribute to the appearance of pancreatic islet autoimmunity and type I diabetes onset . Effects of mitiglinide ( S 21403 ) on Kir6.2/ Q09428 , Kir6.2/SUR2A and Kir6.2/SUR2B types of DB00171 -sensitive potassium channel . 1. We have investigated the mechanism of action of the novel anti-diabetic agent mitiglinide ( S 21403 ) on Kir6.2/ Q09428 , Kir6.2/SUR2A and Kir6.2/SUR2B types of DB00171 -sensitive potassium ( K( DB00171 ) ) channel . These possess a common pore-forming subunit , Kir6.2 , and different regulatory sulphonylurea receptor ( Q09428 ) subunits . It is believed that they correspond to native K( DB00171 ) channels in pancreatic beta-cells , heart and non-vascular smooth muscle , respectively . 2 . Kir6.2 was coexpressed with Q09428 , SUR2A or SUR2B in Xenopus oocytes and macroscopic currents were recorded in giant inside-out membrane patches . DB01252 was added to the intracellular membrane surface . 3 . DB01252 inhibited Kir6.2/ Q09428 currents at two sites : a low-affinity site on Kir6.2 and a high-affinity site on Q09428 . Low-affinity inhibition was similar for all three types of K( DB00171 ) channel but high-affinity inhibition was greater for Kir6.2/ Q09428 currents ( IC(50) , 4 nM ) than for Kir6.2/SUR2A or Kir6.2/SUR2B currents ( IC(50) , 3 and 5 microM , respectively ) . 4 . Inhibition of Kir6.2/ Q09428 currents was only slowly reversible on the time scale of electrophysiological experiments . 5 . Kir6.2/ Q09428 -S1237Y currents , which previously have been shown to lack high affinity tolbutamide inhibition , resembled Kir6.2/SUR2 currents in being unaffected by 100 nM but blocked by 10 microM mitiglinide . 6 . Our results show that mitiglinide is a high-affinity drug that shows a 1000 fold greater affinity for the beta-cell type than the cardiac and smooth muscle types of K( DB00171 ) channel , when measured in excised patches . Synergistic inhibition of breast cancer cell lines with a dual inhibitor of P00533 -HER-2/neu and a Bcl-2 inhibitor . The epidermal growth factor receptor ( P00533 ) ( ErbB1 ) and HER-2/neu ( ErbB2 ) are members of the ErbB family of receptor tyrosine kinases . These receptors are overexpressed in a variety of human tumors and overexpression generally correlates with poor prognosis and decreased survival . DB01259 , a reversible inhibitor of both P00533 and HER-2/neu , has shown some success in achieving clinical responses in heavily pretreated advanced cancer patients . GW2974 is a reversible dual inhibitor similar to lapatinib , but GW2974 was not progressed to clinical trials due to pharmacokinetic issues . Bcl-2 , an anti-apoptotic protein , is also overexpressed in a number of human tumors . Bcl-2 inhibitors induce apoptosis and sensitize cancer cells to other therapies . The purpose of this study was to assess the effects of combining ErbB and Bcl-2 inhibitors on the growth of human breast cancer cell lines . P00533 /HER-2/neu tyrosine kinase inhibitors ( lapatinib and GW2974 ) were combined with Bcl-2 inhibitors ( HA14-1 or GX15-070 ) and the anti-proliferative effects were determined by the MTT tetrazolium dye assay . Combinations were tested in MCF-7 human breast cancer cells , a HER-2/neu transfected MCF-7 cell line ( MCF/18 ) , and a tamoxifen-resistant MCF-7 cell line ( Q99707 -3 ) . A synergistic inhibitory effect was observed with the combination of inhibitors of P00533 -HER-2/neu ( lapatinib or GW2974 ) and Bcl-2 ( GX15-070 or HA14-1 ) on the growth of the MCF-7 , MCF/18 , and Q99707 -3 human breast cancer cell lines . This study suggests that simultaneously blocking the ErbB family of receptor tyrosine kinases and Bcl-2 family of proteins may be a benefit to breast cancer patients . ICE/ P29466 inhibitors as novel anti-inflammatory drugs . In recent years , several strategies that selectively inhibit pro-inflammatory cytokines , have yielded effective protein-based therapies for inflammatory disorders , validating the therapeutic hypothesis that intervention in cytokine signalling can provide clinical benefit . However , these protein-based products must be administered by injection , a constraint associated with inconvenience , adverse effects and expense for patients , caregivers and insurers . Besides interfering with the effects of cytokines such as P01375 or IL-1beta that have already been produced , inhibition of pro-inflammatory cytokine production or signalling with low-molecular weight orally-active drugs would combine the convenience of conventional pharmaceuticals with the focused efficacy of the protein therapies . Reducing IL-1beta and Q14116 production by inhibition of IL-1beta converting enzyme ( ICE , caspase-1 ) is one promising strategy because of the key roles of these cytokines in many inflammatory diseases . DB04875 , the first orally available , potent and selective ICE inhibitor to enter clinical trials , is currently under investigation in rheumatoid arthritis . P19957 kinase/AKT pathway as a therapeutic target in multiple myeloma . The development of novel therapies for multiple myeloma depends on a comprehensive understanding of the events leading to cellular proliferation and survival . Controlling pathways that regulate growth signals is an emerging and complementary approach to myeloma treatment . The PI3K/Akt pathway is a central gatekeeper for crucial cellular functions including adhesion , angiogenesis , migration and development of drug resistance . Established proteins and genes such as P42345 , p53 , NF-kappaB and Q92934 are all regulated through PI3K and Akt activation , making them attractive targets for broad downstream effects . Direct PI3K inhibition has demonstrated impressive tumor inhibition and regression in cell-line and animal models , and multiple agents including DB05210 are currently in clinical trials . Drugs such as perifosine that are specific for Akt are also in development . Combinations of these agents with existing therapies are rational approaches on the path to improving myeloma treatment . 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 . 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 . 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 . 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 . Excitation-transcription coupling via calcium/calmodulin-dependent protein kinase/ P27361 /2 signaling mediates the coordinate induction of Q9P2U8 and Narp triggered by a prolonged increase in glutamatergic synaptic activity . Homeostatic scaling of glutamatergic and GABAergic transmission is triggered by prolonged alterations in synaptic neuronal activity . We have previously described a presynaptic mechanism for synaptic homeostasis and plasticity that involves scaling the level of vesicular glutamate ( Q9P2U7 ) and gamma-aminobutyric acid ( GABA ) ( Q9H598 ) transporter biosynthesis . These molecular determinants of vesicle filling and quantal size are regulated by neuronal activity in an opposite manner and bi-directionally . Here , we report that a striking induction of Q9P2U8 mRNA and synaptic protein is triggered by a prolonged increase in glutamatergic synaptic activity in mature neocortical neuronal networks in vitro together with two determinants of inhibitory synaptic strength , the neuronal activity-regulated pentraxin ( Narp ) , and glutamate decarboxylase ( Q05329 ) . Activity-dependent induction of Q9P2U8 and Narp exhibits a similar intermediate-early gene response that is blocked by actinomycin D and tetrodotoxin , by inhibitors of ionotropic glutamate receptors and L-type voltage-gated calcium channels , and is dependent on downstream signaling via calmodulin , calcium/calmodulin-dependent protein kinase ( CaMK ) and extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . The co-induction of Q9P2U8 and Narp triggered by prolonged gamma-aminobutyric acid type A receptor blockade is independent of brain-derived nerve growth factor and TrkB receptor signaling . Q9P2U8 protein induction occurs on a subset of cortically derived synaptic vesicles in excitatory synapses on somata and dendritic processes of multipolar GABAergic interneurons , recognized sites for the clustering of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate glutamate receptors by Narp . We propose that Q9P2U8 and Narp induction by excitation-transcription coupling leads to increased glutamatergic transmission at synapses on GABAergic inhibitory feedback neurons as part of a coordinated program of Ca(2+)-signal transcription involved in mechanisms of homeostatic plasticity after prolonged hyperactivity . DB00134 recycling pathways and antimalarial drug design . 5'-Deoxy-5'-(methylthio)adenosine ( MTA ) is an S-adenosylmethionine metabolite that is generated as a by-product of polyamine biosynthesis . In mammalian cells , MTA undergoes a phosphorolytic cleavage catalyzed by Q13126 to produce adenine and 5-deoxy-5-(methylthio)ribose-1-phosphate ( Q15012 ) . DB00173 is utilized in purine salvage pathways , and Q15012 is subsequently recycled to methionine . Whereas some microorganisms metabolize MTA to Q15012 via Q13126 , others metabolize MTA to Q15012 in two steps via initial cleavage by MTA nucleosidase to adenine and 5-deoxy-5-(methylthio)ribose ( Q99707 ) followed by conversion of Q99707 to Q15012 by Q99707 kinase . In order to assess the extent to which these pathways may be operative in Plasmodium falciparum , we have examined a series of 5'-alkyl-substituted analogs of MTA and the related Q99707 analogs and compared their abilities to inhibit in vitro growth of this malarial parasite . The Q99707 analogs 5-deoxy-5-(ethylthio)ribose and 5-deoxy-5-(hydroxyethylthio)ribose were inactive at concentrations up to 1 mM , and 5-deoxy-5-(monofluoroethylthio)ribose was weakly active ( 50 % inhibitory concentration = 700 microM ) . In comparison , the MTA analogs , 5'-deoxy-5'-(ethylthio)adenosine,5'-deoxy-5'-(hydroxyethylthio)ade nosine ( HETA ) , and 5'-deoxy-5'-(monofluoroethylthio)adenosine , had 50 % inhibitory concentrations of 80 , 46 , and 61 microM , respectively . Extracts of P. falciparum were found to have substantial Q13126 activity . Coadministration of MTA with HETA partially protected the parasites against the growth-inhibitory effects of HETA . Results of this study indicate that P. falciparum has an active Q13126 that can be targeted by analogs of MTA . Inhibition of PI3K/AKT/ P42345 pathway enhances temozolomide-induced cytotoxicity in pituitary adenoma cell lines in vitro and xenografted pituitary adenoma in female nude mice . Invasive pituitary adenomas ( PAs ) are often refractory to standard therapy and salvage treatment with temozolomide ( DB00853 ) . Hyperactivation of the phosphoinositide 3-kinase ( PI3K ) /AKT/mammalian target of rapamycin ( P42345 ) pathway contributes to chemotherapy resistance in many cancers . DB05241 , a novel dual-PI3K/ P42345 inhibitor , has recently shown its efficacy as a monotherapy and in combination with conventional therapeutics in many cancers . The hyperactive PI3K/AKT/ P42345 pathway frequently occurs in invasive PAs . In this study , we investigated whether DB05241 sensitizes PA cells to DB00853 in vitro and in vivo . Experiments were carried out to evaluate the effect of DB05241 and DB00853 alone or in combination on cell proliferation and apoptosis of PA cell lines ( α DB00279 -1 , GH3 , and MMQ ) in vitro as well as the tumor growth and serum GH and prolactin secretions in a GH3 xenograft tumor model of female nude mice . DB05241 and DB00853 synergistically inhibited the growth of PA cell lines and induced apoptosis . Combination of DB05241 and DB00853 synergistically inhibited tumor growth , decreased serum GH and prolactin levels , and reduced the sacrifice rate of GH3 xenograft tumor models without increased systemic side effects . In addition , DB05241 in combination with DB00853 dramatically decreased phosphorylation of AKT and P42345 as well as the expression of Bcl-2 . The increased expression of cleaved poly ( ADP-ribose ) polymerase and Bcl-2-associated X protein along with elevated caspase-3/7 activity were also observed in the combination group . Therefore , dual inhibitors of PI3K and P42345 may enhance alkylating agent-mediated cytotoxicity and provide a novel regimen in the treatment of invasive PAs . Turning receptors on and off with intracellular pepducins : new insights into G-protein-coupled receptor drug development . G-protein-coupled receptors ( GPCRs ) are a large family of remarkably versatile membrane proteins that are attractive therapeutic targets because of their involvement in a vast range of normal physiological processes and pathological diseases . Upon activation , intracellular domains of GPCRs mediate signaling to G-proteins , but these domains have yet to be effectively exploited as drug targets . Cell-penetrating lipidated peptides called pepducins target specific intracellular loops of GPCRs and have recently emerged as effective allosteric modulators of GPCR activity . The lipid moiety facilitates translocation across the plasma membrane , where pepducins then specifically modulate signaling of their cognate receptor . To date , pepducins and related lipopeptides have been shown to specifically modulate the activity of diverse GPCRs and other membrane proteins , including protease-activated receptors ( PAR1 , PAR2 , and PAR4 ) , chemokine receptors ( P25024 , P25025 , and P61073 ) , sphingosine 1-phosphate receptor-3 ( Q99500 ) , the melanocortin-4 receptor , the Smoothened receptor , formyl peptide receptor-2 ( P25090 ) , the relaxin receptor ( Q9HBX9 ) , G-proteins ( Gα(q/11/o/13) ) , muscarinic acetylcholine receptor and vanilloid ( Q8NER1 ) channels , and the P08514 integrin . This minireview describes recent advances made using pepducin technology in targeting diverse GPCRs and the use of pepducins in identifying potential novel drug targets .

MH_train_1184

MH_train_1184

MH_train_1184

interacts_with DB00850?

multiple_choice

P15121 inhibitor fidarestat attenuates leukocyte-endothelial interactions in experimental diabetic rat retina in vivo . PURPOSE : Dysregulation of the polyol pathway has been implicated as a major cause of diabetic retinopathy . The aldose reductase inhibitor fidarestat was recently reported to prevent retinal oxidative stress and overexpression of vascular endothelial growth factor ( P15692 ) protein in diabetic rats . In this study , we investigated the effect of fidarestat on leukocyte-endothelial cell interactions in an in vivo experimental model for diabetic retina . MATERIALS AND METHODS : Diabetes was induced in six-week-old male Long-Evans rats by intraperitoneal injection of streptozotocin ( Q11206 ) ( 75 mg/kg ) . The rats were divided into four experimental groups : non-diabetic control rats , untreated diabetic rats , and diabetic rats treated with a low ( 4 mg/kg/day ) or high ( 16 mg/kg/day ) oral dose of fidarestat . After four weeks of treatment , accumulated leukocytes in the retina were counted in vivo by acridine orange digital fluorography . Intercellular adhesion molecule-1 ( P05362 ) and P15692 -164 mRNA levels in the retina were analyzed using the quantitative reverse transcription-polymerase chain reaction . P05362 protein expression in the retina was investigated by immunohistochemistry . RESULTS : DB02021 treatment significantly decreased concentrations of sorbitol and fructose in the retinas of Q11206 -induced diabetic rats . Leukocyte accumulation in the retinas of fidarestat-treated rats was significantly less than in the untreated diabetic group ( P < 0.01 ) . DB02021 treatment significantly reduced the expression P05362 mRNA , but not P15692 -164 mRNA , in the retina of diabetic rats . Immunohistochemical study also revealed the suppressive effect of fidarestat on expression of P05362 . CONCLUSIONS : Oral administration of fidarestat attenuated leukocyte accumulation in the retina of Q11206 induced-diabetic rats , suggesting that fidarestat may have a therapeutic role in preventing the progression of diabetic retinopathy . 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 . 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 . 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 . Reduced satiating effect of d-fenfluramine in serotonin 5-HT(2C) receptor mutant mice . RATIONALE : d- DB00574 stimulates the release of serotonin ( 5-HT ) and is a potent inhibitor of the re-uptake of 5-HT into nerve terminals . Administration of d-fenfluramine suppresses food intake in both animals and humans . OBJECTIVE : We have investigated the role of the P28335 receptor in mediating the effect of d-fenfluramine on mouse food intake and the behavioural satiety sequence . METHODS : Mutant mice lacking serotonin P28335 receptors and wild-type animals were habituated to a daily presentation of wet mash . Animals were non-deprived and received d-fenfluramine ( 3-30 mg/kg ) 30 min prior to being assessed for the presence of stereotypy and presented with wet mash . The behaviour of animals was observed for the subsequent 40 min and food intake was recorded . RESULTS : d- DB00574 dose-dependently inhibited the consumption of a palatable wet mash by the mice . d- DB00574 ( 3 mg/kg ) significantly reduced the amount of wet mash consumed by wild-type mice and induced a temporal advance in the behavioural satiety sequence consistent with an enhancement of satiety . Mutant mice were less sensitive to the satiating effects of 3 mg/kg d-fenfluramine . Hence , this dose of d-fenfluramine had a reduced effect on both food consumption and the behavioural satiety sequence in the P28335 mutant mice . In contrast , mutant mice showed an increased sensitivity to the stereotypy induced by high doses of d-fenfluramine ( 10 , 30 mg/kg ) compared to that of wild-type littermates . CONCLUSION : These data demonstrate a role for the P28335 receptor in mediating d-fenfluramine-induced satiety . Sporadic fundic gland polyps : an immunohistochemical study of their antigenic profile . Fundic Gland Polyps ( FGPs ) are small sessile ( 2-5 mm ) , usually multiple polyps arising in the gastric , acid-secreting mucosa of disputed histogenesis . They have been described in a sporadic form , prevalently in middle aged females , or associated with familial adenomatosis coli-Gardner 's syndrome . We performed an immunohistochemical study on 24 sporadic FGPs , using monoclonal antibodies ( MAbs ) against differentiation markers , class II MHC antigens ( HLA-DR ) , oncofetal and proliferation antigens , aimed to characterize the antigenic profile of the polyps . A preliminary cytogenetic study on five polyps was also done , using an in situ culture method after collagenase treatment . Cytokeratins 8-18 ( P62158 5.2 MAb ) and 20 ( IT-Ks 20.8 MAb ) , Epithelial Membrane Antigen ( P15941 ) and Chromogranin A were normally expressed by FGPs . FGPs did not express HLA II DR . FGPs did not react with an anti- P06731 MAb ( F6 ) , but they were frequently positive ( 22/24 , 91.6 % ) with B72.3 MAb ( reacting with the cancer-associated mucin epitope sialyl-Tn ) . The PC10 MAb ( against P12004 or cyclin ) showed enhanced expression in the deep glandular-cystic compartment of FGPs ; the P12004 index of FGPs was significantly higher than in normal fundic mucosa . The cytogenetic study on the 5 cases analysed , revealed a normal karyotype . We have demonstrated that FGPs express in the paranuclear zone the sialyl-Tn epitope , a side-chain sugar normally masqued in adult gastric mucins , thus revealing an alteration in mucin synthesis ; FGPs ' higher proliferation index as compared with normal fundic mucosa supports the hypothesis of their hyperproliferative nature . DB06080 inhibits the proliferation of Ewing Sarcoma cells and suppresses platelet-derived growth factor receptor beta and c- P10721 signaling pathways . The Ewing Sarcoma ( Q01844 ) family of tumors is one of the most common tumors diagnosed in children and adolescents and is characterized by a translocation involving the Q01844 gene . Despite advances in chemotherapy , the prognosis of metastatic Q01844 is poor with an overall survival of < 30 % after 5 years . Q01844 tumor cells express the receptor tyrosine kinases , platelet-derived growth factor receptor ( P09619 ) and c- P10721 . DB06080 is a multitargeted small-molecule inhibitor that targets Fms-like tyrosine kinase-3 , c- P10721 , vascular endothelial growth receptors , and PDGFRs . To determine the potential therapeutic benefit of DB06080 in Q01844 cells , we examined the effects of DB06080 on Q01844 cell lines and xenograft mouse models . DB06080 inhibited the proliferation of two Q01844 cell lines , A4573 and TC71 , at an IC(50) of 1.25 and 2 mumol/L after 72 h of treatment , respectively . The phosphorylation of PDGFRbeta , c- P10721 , and extracellular signal-regulated kinases was also inhibited . To examine the effects of DB06080 in vivo , the drug was given to mice injected with Q01844 cells . We observed inhibition of growth of Q01844 tumor cells in a xenograft mouse model and prolonged survival in a metastatic mouse model of Q01844 . Therefore , our in vitro and in vivo studies show that DB06080 inhibits proliferation of Q01844 cells through inhibition of PDGFRbeta and c- P10721 pathways . The Lyme disease vaccine takes its toll . Vaccination with Borrelia burgdorferi outer surface protein ( Osp ) A can induce a protective response against Lyme disease and serves as a model to understand the generation of protective immune responses against the spirochete . The innate response to pathogens is activated by specific Toll-like receptors ( TLRs ) that recognize distinct pathogen-associated molecular patterns . O60603 is of particular interest for B. burgdorferi research because O60603 recognizes several pathogen-associated molecular patterns , including lipoproteins . O60603 may form heterodimers with Q9Y2C9 to identify diacylated lipoproteins , while O60603 works in concert with Q15399 to recognize triacylated lipoproteins such as OspA . We will discuss the role of Q15399 /2 in the development of responses to OspA in Q15399 -and O60603 -deficient mice , and in selected individuals that received the OspA vaccine . While > 95 % of human OspA-based Lyme disease vaccine recipients develop OspA antibodies , a very small group of individuals did not develop detectable humoral responses against OspA . We demonstrated that this hyporesponsiveness to OspA vaccination was associated with decreased cell surface expression of Q15399 . Moreover , Q15399 - and O60603 -deficient mice did not develop significant levels of OspA antibodies following vaccination with OspA , providing a correlation with human hyporesponsiveness to OspA . These data suggest that defects in the Q15399 /2 signaling pathway are associated with an impaired ability to generate antibodies following immunization with DB00045 . 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 . DB01686 , oxidative stress , and vascular nitric oxide synthase in essential hypertension . We reported impaired endothelium-derived relaxation factor/nitric oxide ( EDRF/NO ) responses and constitutive nitric oxide synthase ( P29474 ) activity in subcutaneous vessels dissected from patients with essential hypertension ( n = 9 ) compared with normal controls ( n = 10 ) . We now test the hypothesis that the patients in this study have increased circulating levels of the P29474 inhibitor , asymmetric dimethylarginine ( DB01686 ) , or the lipid peroxidation product of linoleic acid , 13-hydroxyoctadecadienoic acid ( HODE ) , which is a marker of reactive oxygen species . Patients had significantly ( P < 0.001 ) elevated ( means +/- SD ) plasma levels of DB01686 ( P( DB01686 ) , 766 +/- 217 vs. 393 +/- 57 nmol/l ) and symmetric dimethylarginine ( P(SDMA) : 644 +/- 140 vs. 399 +/- 70 nmol/l ) but similar levels of L-arginine accompanied by significantly ( P < 0.015 ) increased rates of renal DB01686 excretion ( 21 +/- 9 vs. 14 +/- 5 nmol/mumol creatinine ) and decreased rates of renal DB01686 clearance ( 18 +/- 3 vs. 28 +/- 5 ml/min ) . They had significantly increased plasma levels of HODE ( P(HODE) : 309 +/- 30 vs. 226 +/- 24 nmol/l ) and renal HODE excretion ( 433 +/- 93 vs. 299 +/- 67 nmol/micromol creatinine ) . For the combined group of normal and hypertensive subjects , the individual values for plasma levels of DB01686 and HODE were both significantly ( P < 0.001 ) and inversely correlated with microvascular EDRF/NO and positively correlated with mean blood pressure . In conclusion , elevated levels of DB01686 and oxidative stress in a group of hypertensive patients could contribute to the associated microvascular endothelial dysfunction and elevated blood pressure . 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 . Expression of P16070 , P05362 and N- P62158 in colorectal cancer . Correlation with the tumor stage and the phenotypical characteristics of tumor-infiltrating lymphocytes . The cellular adhesion molecules ( CAMs ) CD44s , CD44v6 , CD44v10 , P05362 and N- P62158 were immunohistologically detected in colorectal cancers using the APAAP method . The expression of CD44s and CD44v6 was associated with the presence of lymph node metastases in the examined tumors . The pattern of P05362 expression was inversely related to that of P16070 , i.e. lower numbers of P05362 positive cells were observed in metastasizing tumors . An intense focal staining of N- P62158 was observed in the majority of the metastasizing tumors . The expression of CD44v , P05362 or N- P62158 on tumor cells did not correlate with the density of the tumor-infiltrating lymphocytes ( Q15399 ) within the tumors . The flowcytometric analysis of Q15399 showed a significant accumulation of CD25+ and HLA-DR+ cells and a reduced number of CD45RA+ cells as compared to autologous peripheral blood lymphocytes ( PBL ) or intraepithelial lymphocytes of the colon mucosa ( IEL ) . These phenotypic characteristics of Q15399 did not correlate with the CAMexpression on tumor cells . 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 . DB08875 ( DB05153 ) for the treatment of locally advanced or metastatic progressive medullary thyroid cancer . DB08875 ( DB05153 ) is an oral multiple receptor tyrosine kinase inhibitor manufactured by Exelixis Inc. , CA , USA . It mainly inhibits three tyrosine kinase receptors : MET , P35968 and P07949 . In both preclinical and clinical studies it has been shown to inhibit tumor angiogenesis , invasiveness and metastases . The most frequent side effects are fatigue , diarrhea , decreased appetite , nausea , weight loss and palmar-plantar erythrodysesthesia . A Phase III clinical trial ( EXAM study ) of DB05153 versus placebo in advanced and progressive medullary thyroid cancer showed a 28 versus 0 % overall response rate and a progression-free survival of 11.2 versus 4.0 months ( hazard ratio : 0.28 ; 95 % CI : 0.19-0.40 ; p < 0.0001 ) in patients treated with cabozantinib and placebo , respectively . The drug has been approved by the US FDA for the treatment of advanced/progressive metastatic medullary thyroid cancer in the USA . The P15941 is now evaluating its approval in Europe . P30047 -dependent and -independent inhibitors of P30793 . P30047 ( P30047 ) mediates the feedback inhibition of P30793 activity by ( 6R ) -L-erythro- DB00360 ( BH4 ) through protein complex formation . Since guanine and BH4 have a common pyrimidine ring structure , we examined the inhibitory effect of guanine and its analogs on the enzyme activity . DB02377 , 8-hydroxyguanine , 8-methylguanine , and 8-bromoguanine inhibited the enzyme activity in a P30047 -dependent and pH-dependent manner and induced complex formation between P30793 and P30047 . The type of inhibition by this group is a mixed type . All these properties were shared with BH4 . In striking contrast , inhibition by DB01667 and 8-mercaptoguanine was P30047 -independent and pH-independent . The type of inhibition by DB01667 and 8-mercaptoguanine was a competitive type . The two compounds did not induce complex formation between the enzyme and P30047 . These results demonstrate that guanine compounds of the first group bind to the BH4-binding site of the P30793 / P30047 complex , whereas DB01667 and 8-mercaptoguanine bind to the active site of the enzyme . Finally , the possible implications in Lesch-Nyhan syndrome and Parkinson diseases of the inhibition of P30793 by guanine and 8-hydroxyguanine are discussed . [ Effects of various growth factors on the growth of trophoblast cells in long-term culture ] . Trophoblasts taken from placental tissue of the 1st trimester and molar tissue , and P30793 -1 ( gestational choriocarcinoma cell line ) cells were cultured in collagen coated dishes . The medium used was a mixture of DME and Ham 's F-12 ( 1:1 ) , containing P01133 , PDGF , insulin , GM- P04141 , IL-1 , -2 , -3 , PGE1 and DB00917 in various concentrations . 3H-TdR uptake of the cultured cells was measured as a marker of cell growth . The growth of normal trophoblasts was enhanced by PDGF or insulin , and remarkably by P01133 + PDGF + insulin + GM- P04141 . The growth of molar trophoblasts was accelerated by P01133 or insulin . Under these culture conditions , normal trophoblasts have been successfully cultured and maintained for 12 months and molar trophoblasts for 9 months to date . The cultured cells were identified with trophoblasts by immunohistochemical staining with P62158 5.2 monoclonal antibody . No effect of growth factors was observed in P30793 -1 cells . DB09210 defines a new binding site for allosteric modulators of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid ( AMPA ) receptors . Glutamate receptors are the most prevalent excitatory neurotransmitter receptors in the vertebrate central nervous system and are important potential drug targets for cognitive enhancement and the treatment of schizophrenia . Allosteric modulators of AMPA receptors promote dimerization by binding to a dimer interface and reducing desensitization and deactivation . The pyrrolidine allosteric modulators , piracetam and aniracetam , were among the first of this class of drugs to be discovered . We have determined the structure of the ligand binding domain of the AMPA receptor subtypes P42262 and P42263 with piracetam and a corresponding structure of P42263 with aniracetam . Both drugs bind to P42262 and P42263 in a very similar manner , suggesting little subunit specificity . However , the binding sites for piracetam and aniracetam differ considerably . DB04599 binds to a symmetrical site at the center of the dimer interface . DB09210 binds to multiple sites along the dimer interface with low occupation , one of which is a unique binding site for potential allosteric modulators . This new site may be of importance in the design of new allosteric regulators . 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 . P23560 activation of P62158 -kinase kinase via transient receptor potential canonical channels induces the translation and synaptic incorporation of P42261 -containing calcium-permeable AMPA receptors . Glutamatergic synapses in early postnatal development transiently express calcium-permeable AMPA receptors ( CP-AMPARs ) . Although these P42262 -lacking receptors are essential and are elevated in response to brain-derived neurotrophic factor ( P23560 ) , little is known regarding molecular mechanisms that govern their expression and synaptic insertion . Here we show that P23560 -induced P42261 translation in rat primary hippocampal neurons requires the activation of mammalian target of rapamycin ( P42345 ) via calcium calmodulin-dependent protein kinase kinase ( CaMKK ) . Specifically , P23560 -mediated phosphorylation of threonine 308 ( T308 ) in AKT , a known substrate of CaMKK and an upstream activator of P42345 -dependent translation , was prevented by ( 1 ) pharmacological inhibition of CaMKK with STO-609 , ( 2 ) overexpression of a dominant-negative CaMKK , or ( 3 ) short hairpin-mediated knockdown of CaMKK . P42261 surface expression induced by P23560 , as assessed by immunocytochemistry using an extracellular N-terminal P42261 antibody or by surface biotinylation , was impaired following knockdown of CaMKK or treatment with STO-609 . Activation of CaMKK by P23560 requires transient receptor potential canonical ( TRPC ) channels as SKF-96365 , but not the DB01221 receptor antagonist d-APV , prevented P23560 -induced P42261 surface expression as well as phosphorylation of CaMKI , AKT(T308) , and P42345 . Using siRNA we confirmed the involvement of Q9UL62 and Q9Y210 subunits in P23560 -induced AKT(T308) phosphorylation . The P23560 -induced increase in mEPSC was blocked by IEM-1460 , a selected antagonist of CP-AMPARs , as well as by the specific repression of acute P42261 translation via siRNA to P42261 but not P42262 . Together these data support the conclusion that newly synthesized P42261 subunits , induced by P23560 , are readily incorporated into synapses where they enhance the expression of CP-AMPARs and synaptic strength .

MH_train_1185

MH_train_1185

MH_train_1185

interacts_with DB00864?

multiple_choice

DB04860 , an agonist of Q9NYK1 , reduces plasma virus concentration in chronic hepatitis C infection . Immune-based therapy is the mainstay treatment for chronic hepatitis C virus ( HCV ) infection but causes multiple side effects and achieves durable viral clearance in only approximately 50 % of patients . Most new investigational anti-HCV compounds are direct-acting antivirals for which durability of response and risk of viral mutations and resistance are not yet known . Therefore , continuing discovery and development of new immune-based treatments is desirable . Toll-like receptors ( TLRs ) are pathogen recognition receptors that initiate the innate immune response . The responsiveness of HCV or other ongoing chronic systemic infections to treatment with a selective TLR agonist has not been reported . DB04860 is a selective agonist of Q9NYK1 . In a proof-of-concept study , we found that once-daily 7-day treatment with intravenous isatoribine 800 mg caused a significant ( P = .001 ) reduction of plasma HCV RNA ( mean , -0.76 ; range , -2.85 to +0.21 log(10) units ) in otherwise untreated patients ( n = 12 ) who were chronically infected with HCV . Viral load reduction occurred in patients infected with genotype 1 as well as non-genotype 1 HCV . The reduction of viral load was correlated with induction of markers of a heightened immune antiviral state , including 2'- , 5'- oligoadenylate synthetase levels in whole blood . This treatment was well tolerated , with a low frequency of mild to moderate adverse events . In conclusion , systemic administration of the selective Q9NYK1 agonist isatoribine resulted in dose-dependent changes in immunologic biomarkers and a statistically significant antiviral effect with relatively few and mild side effects . Reduced lymphocyte proliferation and interleukin-2 production in anxiety disorders . OBJECTIVE : The purpose of this study was to examine the effect of anxiety on cell-mediated immunity . METHOD : The subjects consisted of 31 patients with anxiety disorders and 31 normal controls , who were gender-matched . Cell-mediated immune function was measured by the lymphocyte proliferative response to phytohemagglutinin ( PHA ) , P60568 ( P60568 ) production , and natural killer cell activity ( P20366 ) . The extent of anxiety was assessed by the Hamilton rating scale for anxiety and the anxiety subscale of symptom checklist-90 revised ( SCL-90-R ) . RESULTS : The patients with anxiety disorders were significantly lower than the normal controls in lymphocyte proliferative response to PHA and P60568 production . However , there was no significant difference in P20366 between the two groups . Also , no significant correlation was found between the duration of illness or the degree of anxiety and each immune measure in patients with anxiety disorders . CONCLUSIONS : The results suggest a reduced cell-mediated immune function in patients with anxiety disorders , compared with normal controls . These findings also imply that a variety of immune measures should be assessed at the same time in this kind of psychoneuroimmunology research . This would help elucidate the relationship between anxiety and immune function , which has been unclear in most previous research using a single immune measure . Cholecystokinin ( CCK ) stimulates aldosterone secretion from human adrenocortical cells via CCK2 receptors coupled to the adenylate cyclase/protein kinase A signaling cascade . Cholecystokinin ( CCK ) IS a regulatory peptide that acts via two receptor subtypes , P32238 and P32239 . RT-PCR demonstrated the expression of both P32238 and P32239 in the zona glomerulosa ( ZG ) , but not zona fasciculata-reticularis cells of the human adrenal cortex . CCK and the P32239 agonist pentagastrin enhanced basal aldosterone secretion from ZG cells without affecting cortisol production from zona fasciculata-reticularis cells . The aldosterone response to CCK and pentagastrin was suppressed by a P32239 antagonist , but not by a P32238 antagonist . DB00183 evoked a sizeable DB02527 , but not inositol triphosphate , response from ZG cells , whereas CCK plus P32239 antagonist was ineffective . The DB02527 response to pentagastrin was abrogated by P32239 antagonist or the adenylate cyclase inhibitor SQ-22536 , and the aldosterone response was abolished by both SQ-22536 and the protein kinase A inhibitor H-89 . Both CCK and pentagastrin increased steroidogenic acute regulatory protein mRNA expression in ZG cells ; the effect was abrogated by P32239 antagonist . We conclude that CCK exerts secretagogue action on human ZG cells , acting through CCK2-Rs coupled to the adenylate cyclase/protein kinase A signaling cascade , which , in turn , stimulates the expression of steroidogenic acute regulatory protein , the rate-limiting step of steroidogenesis . NO-donor P35354 inhibitors . New nitrooxy-substituted 1,5-diarylimidazoles endowed with P35354 inhibitory and vasodilator properties . A series of NO-donor diarylimidazoles derived from the lead compound DB05095 were synthesized and evaluated for their P35354 inhibitory activity and their stability in whole blood as well as for vasodilator properties . The products are partly transformed into the corresponding alcohols following 24-h incubation in whole blood . All of them display good P23219 / P35354 selectivity , but are less potent than the lead ; a molecular modeling study was carried out to investigate their binding mode . The compounds are also capable of relaxing rat aorta strips precontracted with phenylephrine with a NO-dependent mechanism ; this property could confer reduced cardiotoxicity with respect to traditional P35354 inhibitors . Serotonin promotes tumor growth in human hepatocellular cancer . In addition to its function as a neurotransmitter and vascular active molecule , serotonin is also a mitogen for hepatocytes and promotes liver regeneration . A possible role in hepatocellular cancer has not yet been investigated . Human hepatocellular cancer cell lines Huh7 and HepG2 were used to assess the function of serotonin in these cell lines . Characteristics of autophagy were detected with transmission electron microscopy , immunoblots of microtubule-associated protein light chain 3(LC3) and p62 ( sequestosome 1 ) . Immunoblots of the mammalian target of rapamycin ( P42345 ) and its downstream targets p70S6K and Q13541 were used to investigate signaling pathways of serotonin . Two different animal models served as principle of proof of in vitro findings . Clinical relevance of the experimental findings was evaluated with a tissue microarray from 168 patients with hepatocellular carcinoma . Serotonin promotes tumor growth and survival in starved hepatocellular carcinoma cells . During starvation hepatocellular carcinoma cells exhibited characteristics of autophagy , which disappeared in serotonin-treated cells . DB00877 , an inhibitor of P42345 , is known to induce autophagy . Serotonin could override rapamycin by an P42345 -independent pathway and activate common downstream signals such as p70S6K and Q13541 . In two tumor models of the mouse , inhibition of serotonin signaling consistently impaired tumor growth . Human biopsies revealed expression of the serotonin receptor P41595 , correlating with downstream signals , e.g. , phosphorylated p70S6K and proliferation . CONCLUSION : This study provides evidence that serotonin is involved in tumor growth of hepatocellular cancer by activating downstream targets of P42345 , and therefore serotonin-related pathways might represent a new treatment strategy . P01350 induces sodium-hydrogen exchanger 3 phosphorylation and P42345 activation via a phosphoinositide 3-kinase-/protein kinase C-dependent but AKT-independent pathway in renal proximal tubule cells derived from a normotensive male human . P01350 is natriuretic , but its renal molecular targets and signal transduction pathways are not fully known . In this study , we confirmed the existence of P32239 ( a gastrin receptor ) in male human renal proximal tubule cells and discovered that gastrin induced S6 phosphorylation , a downstream component of the phosphatidylinositol 3 kinase ( P19957 kinase ) -mammalian target of rapamycin pathway . P01350 also increased the phosphorylation of sodium-hydrogen exchanger 3 ( P48764 ) at serine 552 , caused its internalization , and decreased its expression at the cell surface and NHE activity . The phosphorylation of P48764 and S6 was dependent on P19957 kinases because it was blocked by 2 different P19957 -kinase inhibitors , wortmannin and LY294,002 . The phosphorylation of P48764 and S6 was not affected by the protein kinase A inhibitor H-89 but was blocked by a pan-PKC ( chelerythrine ) and a conventional PKC ( cPKC ) inhibitor ( Gö6976 ) ( 10 μM ) and an intracellular calcium chelator , 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid , tetra(acetoxymethyl)-ester , suggesting the importance of cPKC and intracellular calcium in the gastrin signaling pathway . The cPKC involved was probably PKCα because it was phosphorylated by gastrin . The gastrin-mediated phosphorylation of P48764 , S6 , and PKCα was via phospholipase C because it was blocked by a phospholipase C inhibitor , U73122 ( 10 μM ) . The phosphorylation ( activation ) of AKT , which is usually upstream of mammalian target of rapamycin in the classic P19957 kinase-AKT-p70S6K signaling pathway , was not affected , suggesting that the gastrin-induced phosphorylation of P48764 and S6 is dependent on both P19957 kinase and PKCα but not AKT . No indication for a defect in toll-like receptor signaling in patients with hyper-IgE syndrome . Hyper-IgE syndrome is a rare primary immunodeficiency of unknown etiology characterized by recurrent infections of the skin and respiratory system , chronic eczema , elevated total serum IgE , and a variety of associated skeletal symptoms . Recent reports about susceptibility to pyogenic bacterial infections and high IgE levels in patients and animals with defects in toll-like receptor ( TLR ) signaling pathways prompted us to search for TLR signaling defects as an underlying cause of hyper-IgE syndrome . Blood samples from six patients with hyper-IgE syndrome were analyzed for serum cytokine levels , intracellular cytokine production in T cells after stimulation with PMA/ionomycin , and cytokine production from peripheral blood mononuclear cells stimulated by TLR ligands and bacterial products including LPS ( O00206 ) , peptidoglycan ( O60603 ) , PolyIC ( O15455 ) , R848 ( Q9NYK1 /8 ) , CpG-A , and CpG-B ( Q9NR96 ) , zymosan and heat killed Listeria monocytogenes . All results were compared to data from healthy controls . A reduction in P01579 , P60568 , and P01375 producing T cells after PMA stimulation suggested a reduced inflammatory T cell response in patients with hyper-IgE syndrome . Increased serum levels of P05113 indicated a concomitant Th2 shift . However , normal production of cytokines ( P01375 , P05231 , P22301 , IFN-alpha , P02778 ) and upregulation of P42081 on B cells and monocytes after TLR stimulation made a defect in TLR signaling pathways highly unlikely . In summary , our data confirmed an imbalance in T cell responses of patients with hyper-IgE syndrome as previously described but showed no indication for an underlying defect in toll-like receptor signaling . Role of serotonin via P41595 receptors in the reinforcing effects of DB01454 in mice . The amphetamine derivative 3,4-methylenedioxymethamphetamine ( DB01454 , ecstasy ) reverses dopamine and serotonin transporters to produce efflux of dopamine and serotonin , respectively , in regions of the brain that have been implicated in reward . However , the role of serotonin/dopamine interactions in the behavioral effects of DB01454 remains unclear . We previously showed that DB01454 -induced locomotion , serotonin and dopamine release are 5-HT(2B) receptor-dependent . The aim of the present study was to determine the contribution of serotonin and 5-HT(2B) receptors to the reinforcing properties of DB01454 .We show here that 5-HT(2B) ( -/- ) mice do not exhibit behavioral sensitization or conditioned place preference following DB01454 ( 10 mg/kg ) injections . In addition , DB01454 -induced reinstatement of conditioned place preference after extinction and locomotor sensitization development are each abolished by a 5-HT(2B) receptor antagonist ( RS127445 ) in wild type mice . Accordingly , DB01454 -induced dopamine D1 receptor-dependent phosphorylation of extracellular regulated kinase in nucleus accumbens is abolished in mice lacking functional 5-HT(2B) receptors . Nevertheless , high doses ( 30 mg/kg ) of DB01454 induce dopamine-dependent but serotonin and 5-HT(2B) receptor-independent behavioral effects.These results underpin the importance of 5-HT(2B) receptors in the reinforcing properties of DB01454 and illustrate the importance of dose-dependent effects of DB01454 on serotonin/dopamine interactions . 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 . 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 . Roles of P09917 and cysteinyl-leukotriene type 1 receptors in the hematological response to allergen challenge and its prevention by diethylcarbamazine in a murine model of asthma . DB00711 ( DEC ) , which blocks leukotriene production , abolishes the challenge-induced increase in eosinopoiesis in bone-marrow from ovalbumin- ( OVA- ) sensitized mice , suggesting that P09917 ( P09917 ) products contribute to the hematological responses in experimental asthma models . We explored the relationship between P09917 , central and peripheral eosinophilia , and effectiveness of DEC , using DB00233 or BALB/c mice and P09917 -deficient mutants . We quantified eosinophil numbers in freshly harvested or cultured bone-marrow , peritoneal lavage fluid , and spleen , with or without administration of leukotriene generation inhibitors ( DEC and MK886 ) and cisteinyl-leukotriene type I receptor antagonist ( montelukast ) . The increase in eosinophil numbers in bone-marrow , observed in sensitized/challenged wild-type mice , was abolished by MK886 and DEC pretreatment . In ALOX mutants , by contrast , there was no increase in bone-marrow eosinophil counts , nor in eosinophil production in culture , in response to sensitization/challenge . In sensitized/challenged ALOX mice , challenge-induced migration of eosinophils to the peritoneal cavity was significantly reduced relative to the wild-type DB00233 controls . DEC was ineffective in ALOX mice , as expected from a mechanism of action dependent on P09917 . In BALB/c mice , challenge significantly increased spleen eosinophil numbers and DEC treatment prevented this increase . Overall , P09917 appears as indispensable to the systemic hematological response to allergen challenge , as well as to the effectiveness of DEC . 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 . [ P08473 activity in the guinea pig model of asthma ] . P08473 exists in airway epithelial cells , smooth muscle , and submucosa near glands , and cleaves tachykinins to inactive metabolites , thereby reducing there effects . To study the role of enkephalinase in asthmatic response , we measured its activity in guinea pig model of asthma . When compared with the control values , the enkephalinase activity was reduced during in immediate asthmatic response ( Q92932 ) and late asthmatic response ( P10586 ) . Compared with the control values ( 100 % ) , each value was 79.7 % , 73.4 % in the trachea and 74.3 % , 55.7 % in the lung respectively . Tracheal muscle preparation taken from the control , Q92932 , and P10586 groups were made and mounted in oxygenated modified Krebs-Ringer solution . The response was monitored by isometric transducer . Concentration response curves to P20366 with or without phosphoramidon were obtained . The contractile responses of the P10586 groups were enhanced in potency and efficiency . DB02557 potentiated the P20366 induced contraction of control and the Q92932 groups but was less potent in enhancing the contractile response in the P10586 group , showing less enkephalinase activity in the P10586 . These results suggest that the enkephalinase plays an important role in P10586 . In P10586 , the enkephalinase activity may be inhibited and the responsiveness of the smooth muscle to some bronchoconstrictor , such as tachykinins , may be increased . 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 . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . [ P08709 -- new physiopathological and therapeutic aspects ] . Factor VII ( FVII ) is a vitamin K dependent coagulation factor that is synthesized in the liver , where clearance of the unactivated protein also takes place . It is a glycoprotein that following activation plays an important role in initiating coagulation after complex formation with tissue factor . A revised hypothesis of blood coagulation implicating the requirement of intact extrinsic and intrinsic pathways is presented . Increased activity of factor VII is associated with atherogenesis , and FVII deficiency may cause bleeding disorders . Recombinant FVIIa ( DB00036 ) may be used in the treatment of haemophilic patients with antibodies against factor VIII or factor IX , utilizing its direct action in activation of factor X . Ongoing studies are investigating whether DB00036 can shorten the bleeding time in patients with thrombocytopenia . 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 . NOS Inhibition Modulates Immune Polarization and Improves Radiation-Induced Tumor Growth Delay . DB00435 synthases ( NOS ) are important mediators of progrowth signaling in tumor cells , as they regulate angiogenesis , immune response , and immune-mediated wound healing . Ionizing radiation ( IR ) is also an immune modulator and inducer of wound response . We hypothesized that radiation therapeutic efficacy could be improved by targeting NOS following tumor irradiation . Herein , we show enhanced radiation-induced ( 10 Gy ) tumor growth delay in a syngeneic model ( C3H ) but not immunosuppressed ( Nu/Nu ) squamous cell carcinoma tumor-bearing mice treated post-IR with the constitutive NOS inhibitor N(G)-nitro-l-arginine methyl ester ( L-NAME ) . These results suggest a requirement of T cells for improved radiation tumor response . In support of this observation , tumor irradiation induced a rapid increase in the immunosuppressive Th2 cytokine P22301 , which was abated by post-IR administration of L-NAME . In vivo suppression of P22301 using an antisense P22301 morpholino also extended the tumor growth delay induced by radiation in a manner similar to L-NAME . Further examination of this mechanism in cultured Jurkat T cells revealed L-NAME suppression of IR-induced P22301 expression , which reaccumulated in the presence of exogenous NO donor . In addition to L-NAME , the guanylyl cyclase inhibitors ODQ and thrombospondin-1 also abated IR-induced P22301 expression in Jurkat T cells and Q14201 -1 macrophages , which further suggests that the immunosuppressive effects involve P29474 . Moreover , cytotoxic Th1 cytokines , including P60568 , IL12p40 , and IFNγ , as well as activated CD8(+) T cells were elevated in tumors receiving post-IR L-NAME . Together , these results suggest that post-IR NOS inhibition improves radiation tumor response via Th1 immune polarization within the tumor microenvironment . Potent and selective inhibition of human nitric oxide synthases . Inhibition by non-amino acid isothioureas . DB02539 was a potent competitive inhibitor of human nitric oxide synthase ( NOS ) , with Ki values of 17 , 36 , and 29 nM for the inducible ( i ) , endothelial ( e ) , and neuronal ( n ) isozymes , respectively . Unlike some potent inhibitors of NOS , no time dependence was observed . DB02539 was not a detectable substrate for P29474 . DB02539 was also a potent inhibitor of mouse P35228 ( Ki value of 5.2 nM ) , and its binding perturbed the spectrum of P35228 consistent with its altering the environment of the bound heme . The optimum binding of S-ethyl- and S-isopropylisothiourea relative to 70 other analogs suggested that these alkyl substitutions fit into a small hydrophobic pocket . Most isothioureas were 2-6-fold selective for the human P35228 ( Ki for P35228 versus Ki for P29474 ) , with one being 19-fold selective . The cyclized mimics of S-ethylisothiourea , 2-NH2-thiazoline , and 2-NH2-thiazole , were also competitive inhibitors of human NOS . A third structural class of inhibitors , bisisothioureas , were , in general , the most selective in their inhibition of human P35228 . S,S'-(1,3-Phenylenebis(1,2-ethanediyl))bisisothiourea was 190-fold selective ( Ki value of 0.047 microM against P35228 versus 9.0 microM against P29474 ) . These results demonstrate that potent and selective inhibition of human NOS isozymes is achievable . Transcriptional regulation of the mouse interleukin-2 receptor beta chain gene by Ets and Egr-1 . To clarify the mechanisms and factors involved in the regulation of mouse IL-2Rbeta gene expression , we isolated the 5'-flanking region of IL-2Rbeta gene and investigated the promoter activity . Here we elucidated the positive regulatory regions , the most potent of which are located between -50 to -30bp and -164 to -135bp . These regions contain a potentially functional Ets and Egr-1-binding sites whose mutations abrogate promoter activity . Data from electrophoretic mobility shift assay indicate that Ets and Egr-1 , but not Sp1 , bind to the positive regulatory regions , -50 to -30bp and -164 to -135bp , respectively . Furthermore , recruitment of Ets and Egr-1 at endogenous IL-2Rbeta promoter segments in an P60568 -dependent P08709 cells was verified by the chromatin immunoprecipitation assay . This study for the first time delineates the molecular mechanisms underlying regulation of mouse IL-2Rbeta gene transcription by Ets family proteins , partially with Egr-1 , and thereby further elucidates the molecular basis of lymphocyte activation and differentiation .

MH_train_1186

MH_train_1186

MH_train_1186

interacts_with DB00758?

multiple_choice

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 . 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 . Q96EB6 activation confers neuroprotection in experimental optic neuritis . PURPOSE : Axonal damage and loss of neurons correlate with permanent vision loss and neurologic disability in patients with optic neuritis and multiple sclerosis ( MS ) . Current therapies involve immunomodulation , with limited effects on neuronal damage . The authors examined potential neuroprotective effects in optic neuritis by SRT647 and DB05073 , two structurally and mechanistically distinct activators of Q96EB6 , an enzyme involved in cellular stress resistance and survival . METHODS : Experimental autoimmune encephalomyelitis ( EAE ) , an animal model of MS , was induced by immunization with proteolipid protein peptide in SJL/J mice . Optic neuritis developed in two thirds of eyes with significant retinal ganglion cell ( RGC ) loss detected 14 days after immunization . RGCs were labeled in a retrograde fashion with fluorogold by injection into superior colliculi . Optic neuritis was detected by inflammatory cell infiltration of the optic nerve . RESULTS : Intravitreal injection of Q96EB6 activators 0 , 3 , 7 , and 11 days after immunization significantly attenuated RGC loss in a dose-dependent manner . This neuroprotective effect was blocked by sirtinol , a Q96EB6 inhibitor . Treatment with either Q96EB6 activator did not prevent EAE or optic nerve inflammation . A single dose of DB05073 on day 11 was sufficient to limit RGC loss and to preserve axon function . CONCLUSIONS : Q96EB6 activators provide an important potential therapy to prevent the neuronal damage that leads to permanent neurologic disability in optic neuritis and MS patients . Intravitreal administration of Q96EB6 activators does not suppress inflammation in this model , suggesting that their neuroprotective effects will be additive or synergistic with current immunomodulatory therapies . Natriuretic peptides induce weak P50552 phosphorylation at DB00133 239 in platelets . Cyclic guanosine-3',5'-monophoshate ( cGMP ) is the common second messenger for the cardiovascular effects of nitric oxide ( NO ) and natriuretic peptides ( NP ; e.g. atrial NP [ P01160 ] ) , which activate soluble and particulate guanylyl cyclases , respectively . The role of NO in regulating cGMP and platelet function is well documented , whereas there is little evidence supporting a role for NPs in regulating platelet reactivity . By studying platelet aggregation and secretion in response to a P25116 peptide , collagen and ADP , and phosphorylation of the cGMP-dependent protein kinase ( PKG ) substrate vasodilator-stimulated phosphoprotein ( P50552 ) at serine 239 , we evaluated the effects of NPs in the absence or presence of the non-selective cGMP and DB02527 phosphodiesterase ( PDE ) inhibitor , DB07954 ( DB07954 ) . Our results show that NPs , possibly through the clearance receptor ( natriuretic peptide receptor-C ) expressed on platelet membranes , increase P50552 phosphorylation but only following PDE inhibition , indicating a small , localised cGMP synthesis . As platelet aggregation and secretion measured under the same conditions were not affected , we conclude that the magnitude of PKG activation achieved by NPs in platelets per se is not sufficient to exert functional inhibition of platelet involvement in haemostasis . The Evi1 proto-oncoprotein blocks endomitosis in megakaryocytes by inhibiting sustained cyclin-dependent kinase 2 catalytic activity . The 3q21q26 syndrome leukaemias are characterised by dystrophic megakaryocytes , elevated platelet counts , ectopic EVI1 protein production and poor prognosis . To investigate the molecular basis of this disease , we developed a model system to examine the biological activity of EVI1 in a megakaryocyte progenitor cell line . For this purpose , Evi1 was conditionally expressed in human erythroleukaemia cells ( HEL ) that progress along the megakaryocyte lineage in the presence of 12-O-tetradecanoylphorbol 13-acetate ( TPA ) . TPA-stimulated HEL cells normally undergo : ( 1 ) growth arrest ; ( 2 ) altered morphology ; ( 3 ) endomitosis and ( 4 ) characteristic changes in gene expression , including reduction of the erythroid-specific glycophoryn A and elevation of the specific glycoproteins P05106 and Q9HCN6 . Enforced Evi1 expression alone had no effect upon HEL cell proliferation or differentiation but a phenotype was manifest upon stimulation to differentiate . Evi1-expressing , TPA-treated HEL cells still showed growth arrest , had reduced and enhanced glycophoryn A and P05106 mRNA 's , respectively , but failed to significantly elevate Q9HCN6 mRNA . This was accompanied by inhibition of endomitosis and altered cell morphology . Sustained P24941 catalytic activity , typically associated with megakaryocyte endomitosis , was dramatically decreased in TPA-stimulated Evi1-expressing HEL cells because of significantly reduced levels of cyclin A . Therefore , enforced Evi1 expression could inhibit megakaryocyte differentiation although retention of some characteristic molecular changes , in combination with a block in endomitosis and altered morphology , suggest a defect in lineage progression . These results suggest that ectopic Evi1 expression contributes to a defective megakaryocyte differentiation programme and is likely to contribute to the phenotype observed in 3q21q26 syndrome leukaemias . Roles of purinergic receptor P2Y , G protein-coupled 12 in the development of atherosclerosis in apolipoprotein E-deficient mice . OBJECTIVE : The aim of the study was to evaluate the role of purinergic receptor P2Y , G protein-coupled 12 ( Q9H244 ) , an ADP receptor , in the development of atherosclerotic lesions . METHODS AND RESULTS : P02649 -null mice were crossed with P2y12(-/-) mice to generate double knockout mice . The double knockout mice and the control apolipoprotein E-null mice were fed a high-fat diet for 20 weeks . Assessment of the atherosclerotic lesions in the control and double knockout mice demonstrated that Q9H244 deficiency caused a diminished lesion area , an increased fibrous content at the plaque site , and decreased monocyte/macrophage infiltration of the lesions . Polymerase chain reaction studies revealed that white blood cells do not express significant levels of Q9H244 . Bone marrow transplantation experiments confirmed that Q9H244 expressed on platelets is a key factor responsible for atherosclerosis , but do not exclude a role of smooth muscle cell Q9H244 . Supernatant fluid from activated P2y12(+/+) but not P2y12(-/-) platelets was capable of causing monocyte migration . In vitro studies showed that platelet Q9H244 deficiency suppressed platelet factor 4 secretion and P16109 expression . Further work demonstrated that platelet Q9H244 , through inhibition of the DB02527 /protein kinase A pathway , critically regulates the release of platelet factor 4 , and thereby affects monocyte recruitment and infiltration . CONCLUSIONS : These results demonstrate that Q9H244 modulates atherogenesis , at least in part by augmenting inflammatory cell recruitment via regulation of platelet α-granule release . P33261 polymorphism is associated with reduced clopidogrel response in cerebrovascular disease . PURPOSE : DB00758 is a prodrug that requires transformation into an active metabolite by cytochrome P450 ( CYP ) in the liver in order to irreversibly inhibit the Q9H244 adenosine diphosphate platelet receptor . P33261 polymorphism has been reported to correlate with reduced antiplatelet activity of clopidogrel in coronary artery disease . We assessed the association between P33261 polymorphism and clopidogrel resistance in patients with cerebrovascular disease . MATERIALS AND METHODS : We retrospectively gathered data from patients who experienced cerebrovascular disease , received clopidogrel , and were tested for clopidogrel resistance and P33261 polymorphism . DB00758 resistance was tested by the VerifyNow Q9H244 system , and the P33261 polymorphism was tested by the Seeplex P33261 P12821 Genotyping system . DB00758 resistance was expressed in Q9H244 reaction units ( PRU ) and percent inhibition . High PRU and low percent inhibition suggests clopidogrel resistance . P33261 polymorphisms were expressed as extensive , intermediate , and poor metabolizers . DB00758 resistance was assessed according to the subgroup of P33261 polymorphism . RESULTS : A total of 166 patients were evaluated . The PRU values of extensive P33261 metabolizers ( 195.0±84.9 ) were significantly lower than those of intermediate and poor metabolizers ( 237.9±88.0 , 302.2±58.9 ) . The percent inhibition of extensive metabolizers ( 44.6±21.8 ) was significantly higher than that of intermediate and poor metabolizers ( 30.5±21.5 , 14.0±13.4 ) . CONCLUSION : Intermediate and poor metabolizing P33261 polymorphism is associated with reduced clopidogrel antiplatelet activity in patients with cerebrovascular disease . The clinical implications of this finding require further investigation . Risk of meningioma and common variation in genes related to innate immunity . BACKGROUND : The etiology of meningioma , the second most common type of adult brain tumor in the United States , is largely unknown . Prior studies indicate that history of immune-related conditions may affect the risk of meningioma . METHODS : To identify genetic markers for meningioma in genes involved with innate immunity , we conducted an exploratory association study of 101 meningioma cases and 330 frequency-matched controls of European ancestry using subjects from a hospital-based study conducted by the National Cancer Institute . We genotyped 1,407 " tag " single nucleotide polymorphisms ( SNP ) in 148 genetic regions chosen on the basis of an r2 > 0.8 and minor allele frequency of > 5 % in Caucasians in HapMap1 . Risk of meningioma was estimated by odds ratios and 95 % confidence intervals . RESULTS : Seventeen SNPs distributed across 12 genetic regions ( P19838 ( 3 ) , P30273 ( 3 ) , CCR6 ( 2 ) , P19320 , P08571 , Q9Y5U5 , P15153 , P47989 , Q13901 , Q15399 / Q9BXR5 / Q9Y2C9 , NOS1 , and Q01523 ) were associated with the risk of meningioma with P < 0.01 . Although individual SNP tests were not significant after controlling for multiple comparisons , gene region-based tests were statistically significant ( P < 0.05 ) for Q9Y5U5 , P19838 , P30273 , P08571 , Q13901 , CCR6 , and P19320 . CONCLUSIONS AND Q9P2X3 : Our results indicate that common genetic polymorphisms in innate immunity genes may be associated with risk of meningioma . Given the small sample size , replication of these results in a larger study of meningioma is needed . Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress . Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis . Because oxidative stress contributes to atherosclerosis , we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon . Bovine aortic endothelial cells were exposed to static , laminar ( 15 dyn/cm2 ) , and oscillatory shear stress ( +/-15 dyn/cm2 ) . Oscillatory shear increased superoxide ( O2.- ) production by more than threefold over static and laminar conditions as detected using electron spin resonance ( P03372 ) . This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources . DB05262 also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence . DB02134 -dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress . This was associated with decreased xanthine dehydrogenase ( P47989 ) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase ( XO ) to P47989 . We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase . These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity . Transfection of these cells with p47phox restored XO protein levels . Finally , in bovine aortic endothelial cells , prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress . These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress . P20711 inhibition does not influence the diuretic and natriuretic response to exogenous alpha-atrial natriuretic peptide in man . The role of dopamine synthesis in the renal actions of human alpha-atrial natriuretic peptide ( alpha P01160 ) was investigated in six dehydrated volunteers using the P20711 inhibitor carbidopa . Each subject received oral placebo or carbidopa ( 100 mg ) followed by an infusion of alpha P01160 10 pmol.kg-1.min-1 for 1 h . The responses to placebo alone and to carbidopa alone were investigated on separate occasions. alpha P01160 produced a similar increase in plasma immunoreactive alpha P01160 whether placebo or carbidopa pretreatment had been given . Urinary dopamine excretion was increased by alpha P01160 . DB00190 pretreatment substantially attenuated this increase without affecting the natriuretic or water-diuretic response to alpha P01160 . DB00190 also failed to alter the change in filtration fraction produced by alpha P01160 . The results suggest that increased synthesis of intrarenal dopamine is not required for the renal effects of alpha P01160 in man . High resolution physical mapping of human O15379 , a potential tumor suppressor gene in the 5q31 region . Histone deacetylases have been described as crucial cofactors of mammalian transcriptional complexes . We have recently identified human histone deacetylase O15379 on chromosome 5q31 by fluorescence in situ hybridization ( Q5TCZ1 ) in a region commonly deleted in malignant myeloid disease . Since O15379 carries strong potential to be a tumor suppressor gene , we report herein its exact position between the P08571 and P42261 genes within the 5q31.1 subband . 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 . The formation of monocyte-platelet aggregates is independent of on-treatment residual agonists'-inducible platelet reactivity . BACKGROUND : Circulating monocyte-platelet aggregates ( DB00603 ) are a sensitive marker of in vivo platelet activation and patients with atherosclerotic vascular disease exhibit higher levels of DB00603 . DB00758 has been shown to reduce DB00603 formation in these patients to a greater extent than aspirin . However , response to clopidogrel and aspirin shows a wide variability , and patients with high on-treatment residual platelet reactivity are at an increased risk for adverse events after coronary stenting . We therefore investigated the association of DB00603 with on-treatment residual agonists'-inducible platelet aggregation in 125 patients on dual antiplatelet therapy after peripheral , coronary or carotid artery stenting . METHODS : DB00603 were characterized by co-expression of monocyte marker P08571 and platelet-specific markers ( CD42b and CD62P ) by whole blood flow cytometry . Platelet reactivity was determined by light transmission aggregometry , the VerifyNow Q9H244 and aspirin assays , and the vasodilator-stimulated phosphoprotein phosphorylation assay . Cut-off values for residual platelet reactivity were defined according to quartiles of each assay . RESULTS : The extent of DB00603 formation showed no significant differences between patients without and with residual ADP-inducible platelet reactivity , and between individuals without and with residual arachidonic acid ( AA ) -inducible platelet reactivity . Even patients with combined on-treatment residual ADP- and AA-inducible platelet reactivity did not exhibit significantly higher levels of DB00603 than patients without any on-treatment residual platelet reactivity . CONCLUSION : High on-treatment residual agonists'-inducible platelet reactivity results in less than a 25 % increase in circulating DB00603 , suggesting that DB00603 formation is largely dependent on other factors . Pelvic fracture mechanism of injury in vehicular trauma patients . To investigate the correlation between motor vehicle crash mechanisms and pelvic injury in front-seat occupants , we retrospectively reviewed the clinical records of , and had complete crash reconstructions performed for , 145 vehicular trauma patients with injury Severity Scores greater than 16 admitted to a level I trauma center . After excluding rear-seat and ejected occupants , 44 of the remaining 115 patients had pelvic injuries . We excluded acetabular fractures and classified the remaining 26 pelvic ring fractures by the system of Young and Burgess : 20 lateral compression ( LC ) fractures , five anteroposterior compression ( P25054 ) fractures , and one combined mechanical injury ( CMI ) fracture . Eighteen pelvic fractures were managed conservatively ; eight required surgical intervention and four of those eight required emergent application of an external fixator for unresponsive hypotension . Trained investigation teams conducted the crash reconstructions , evaluating crash sites and vehicles with direct measurements of more than 500 variables . Calculations from these data , e.g. , direction of impact and change in velocity at impact ( delta V ) , were made with the Q7L266 III computer program and statistical analyses were performed using Chi-square and t tests . This information was then merged with the orthopedic evaluations. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . Epigenetic regulation of angiotensin-converting enzyme 2 ( Q9BYF1 ) by Q96EB6 under conditions of cell energy stress . Q9BYF1 ( angiotensin-converting enzyme 2 ) counterbalances the actions of P12821 ( angiotensin-converting enzyme ) by metabolizing its catalytic product , the vasoactive and fibrogenic peptide AngII ( angiotensin II ) , into Ang-(1-7) [ angiotensin-(1-7) ] . Enhanced Q9BYF1 expression may be protective in diabetes , cardiovascular disease and cancer . However , relatively little is known about the specific physiological factors regulating Q9BYF1 expression . In the present paper , we show , by Western blotting and qPCR ( quantitative real-time PCR ) , that Q9BYF1 expression is increased under conditions of cell stress , including hypoxic conditions , IL (interleukin)-1β treatment and treatment with the AMP mimic AICAR ( 5-amino-4-imidazolecarboxamide riboside ) . The NAD+-dependent deacetylase Q96EB6 ( silent information regulator T1 ) was found to be up-regulated after AICAR treatment but , conversely , was down-regulated after IL-1β treatment . ChIP analysis demonstrated that Q96EB6 bound to the Q9BYF1 promoter and that binding was increased after AICAR treatment , but decreased after IL-1β treatment . Inhibition of Q96EB6 activity ablated the AICAR-induced increase in Q9BYF1 . In conclusion , we have established that the expression of the Q9BYF1 transcript is controlled by the activity of Q96EB6 under conditions of energy stress . Functional significance of DB00142 -77 and DB00135 -137 within the active site of isoaspartyl dipeptidase . Q7L266 ( IAD ) is a binuclear metalloenzyme and a member of the amidohydrolase superfamily . This enzyme catalyzes the hydrolytic cleavage of beta-aspartyl dipeptides . The pH-rate profiles for the hydrolysis of beta- DB00128 - DB00149 indicates that catalysis is dependent on the ionization of two groups ; one that ionizes at a pH approximately 6 and the other approximately 9 . The group that must be ionized for catalysis is directly dependent on the identity of the metal ion bound to the active site . This result is consistent with the ionization of the hydroxide that bridges the two divalent cations . In addition to the residues that interact directly with the divalent cations there are two other residues that are highly conserved and found within the active site : DB00142 -77 and DB00135 -137 . Mutation of DB00135 -137 to phenylalanine reduced the rate of catalysis by three orders of magnitude . The three dimensional X-ray structure of the Y137F mutant did not show any significant conformation changes relative to the three dimensional structure of the wild-type enzyme . The positioning of the side-chain phenolic group of DB00135 -137 in the active site of IAD is consistent with the stabilization of the tetrahedral adduct concomitant with nucleophilic attack by the hydroxide that bridges the two divalent cations . Mutation of DB00142 -77 resulted in the reduction of catalytic activity by five orders of magnitude . The three dimensional structure of the E77Q mutant did not show any significant conformational changes in the mutant relative to the three dimensional structure of the wild-type enzyme . The positioning of the side-chain carboxylate of DB00142 -77 is consistent with the formation of an ion pair interaction with the free alpha-amino group of the substrate . 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 . Comprehensive dissection of PDGF- P09619 signaling pathways in P09619 genetically defined cells . Despite the growing understanding of pdgf signaling , studies of pdgf function have encountered two major obstacles : the functional redundancy of PDGFRalpha and PDGFRbeta in vitro and their distinct roles in vivo . Here we used wild-type mouse embryonic fibroblasts ( MEF ) , MEF null for either PDGFRalpha , beta , or both to dissect PDGF- P09619 signaling pathways . These four P09619 genetically defined cells provided us a platform to study the relative contributions of the pathways triggered by the two PDGF receptors . They were treated with DB00102 and analyzed for differential gene expression , in vitro proliferation and differential response to pharmacological effects . No genes were differentially expressed in the double null cells , suggesting minimal receptor-independent signaling . Protean differentiation and proliferation pathways are commonly regulated by PDGFRalpha , PDGFRbeta and PDGFRalpha/beta while each receptor is also responsible for regulating unique signaling pathways . Furthermore , some signaling is solely modulated through heterodimeric PDGFRalpha/beta . DB09302 inhibits atherosclerosis , improves the plaque morphology , and enhances the effects of a statin . Q8NBP7 ( Q8NBP7 ) inhibition is a potential novel strategy for treatment of CVD . DB09302 is a fully human Q8NBP7 monoclonal antibody in phase 3 clinical development . We evaluated the antiatherogenic potential of alirocumab in P02649 *3Leiden. P11597 mice . Mice received a Western-type diet and were treated with alirocumab ( 3 or 10 mg/kg , weekly subcutaneous dosing ) alone and in combination with atorvastatin ( 3.6 mg/kg/d ) for 18 weeks . DB09302 alone dose-dependently decreased total cholesterol ( -37 % ; -46 % , P < 0.001 ) and TGs ( -36 % ; -39 % , P < 0.001 ) and further decreased cholesterol in combination with atorvastatin ( -48 % ; -58 % , P < 0.001 ) . DB09302 increased hepatic P01130 protein levels but did not affect hepatic cholesterol and TG content . Fecal output of bile acids and neutral sterols was not changed . DB09302 dose-dependently decreased atherosclerotic lesion size ( -71 % ; -88 % , P < 0.001 ) and severity and enhanced these effects when added to atorvastatin ( -89 % ; -98 % , P < 0.001 ) . DB09302 reduced monocyte recruitment and improved the lesion composition by increasing the smooth muscle cell and collagen content and decreasing the macrophage and necrotic core content . DB09302 dose-dependently decreases plasma lipids and , as a result , atherosclerosis development , and it enhances the beneficial effects of atorvastatin in P02649 *3Leiden. P11597 mice . In addition , alirocumab improves plaque morphology . 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 . Novel pyrrolyllactone and pyrrolyllactam indolinones as potent cyclin-dependent kinase 2 inhibitors . P12004 -dependent kinases ( CDKs ) are essential in the control of cell cycle progression . Inhibition of CDKs represents a new approach for pharmacological intervention in the treatment of a variety of proliferative diseases , especially cancer . Based on the crystal structure of P24941 in complex with an imidazole indolinone compound 1 ( DB03428 ) , lead optimization through modeling , synthesis , and SAR studies has led to the discovery of a novel series of pyrrolyllactone and pyrrolyllactam indolinones as potent P24941 inhibitors . PDE10 inhibition increases P42261 and CREB phosphorylation and improves spatial and recognition memories in a Huntington 's disease mouse model . Huntington 's disease ( HD ) causes motor disturbances , preceded by cognitive impairment , in patients and mouse models . We showed that increased hippocampal DB02527 -dependent protein kinase ( PKA ) signaling disrupts recognition and spatial memories in R6 HD mouse models . However , unchanged levels of hippocampal phosphorylated ( p ) DB02527 -responsive element-binding protein ( CREB ) suggested unaltered nuclear PKA activity in R6 mice . Here , we extend this finding by showing that nuclear pPKA catalytic subunit ( Thr197 ) and pPKA substrate levels were unaltered in the hippocampus of R6/1 mice . Phosphodiesterases ( PDEs ) play an important role in the regulation of PKA activity . Q9Y233 , a DB02527 /cGMP dual-substrate PDE , was reported to be restricted to the nuclear region in nonstriatal neurons . Using cell fractionation we confirmed that Q9Y233 was enriched in nuclear fractions , both in wild-type and R6/1 mice hippocampus , without differences in its levels or intracellular distribution between genotypes . We next investigated whether inhibition of PDE10 with papaverine could improve cognitive function in HD mice . DB01113 treatment improved spatial and object recognition memories in R6/1 mice , and significantly increased pGluA1 and pCREB levels in R6/1 mice hippocampus . DB01113 likely acted through the activation of the PKA pathway as the phosphorylation level of distinct cGMP-dependent kinase ( cGK ) substrates was not modified in either genotype . Moreover , hippocampal DB02527 , but not cGMP , levels were increased after acute papaverine injection . Our results show that inhibition of PDE10 improves cognition in R6 mice , at least in part through increased P42261 and CREB phosphorylation . Thus , PDE10 might be a good therapeutic target to improve cognitive impairment in HD . Protease-activated receptor-1 and platelet-derived growth factor in spinal cord neurons are implicated in neuropathic pain after nerve injury . Recently , it has been reported that both thrombin-sensitive protease-activated receptor 1 ( P25116 ) and platelet-derived growth factor ( PDGF ) are present not only in platelets , but also in the CNS , which indicates that they have various physiological functions . In this study , we evaluated whether P25116 /PDGF in the spinal cord could contribute to the development of a neuropathic pain-like state in mice . Thermal hyperalgesia and tactile allodynia induced by sciatic nerve ligation were significantly suppressed by repeated intrathecal injection of hirudin , which is characterized as a specific and potent thrombin inhibitor . Furthermore , a single intrathecal injection of thrombin produced long-lasting hyperalgesia and allodynia , and these effects were also inhibited by hirudin in normal mice . In nerveligated mice , the increase in the binding of [35S]GTPgammaS to membranes of the spinal cord induced by thrombin and P25116 -like immunoreactivity ( IR ) in the spinal cord were each greater than those in sham-operated mice . Thermal hyperalgesia and tactile allodynia induced by sciatic nerve ligation were also suppressed by repeated intrathecal injection of either the PDGF alpha receptor ( PDGFRalpha ) /Fc chimera protein or the P09619 -dependent tyrosine kinase inhibitor AG17 [ (3,5-di-tert-butyl-4-hydroxybenzylidene)-malononitrile ] . Moreover , thermal hyperalgesia and tactile allodynia induced by thrombin in normal mice were virtually eliminated by intrathecal pretreatment with PDGFRalpha/Fc . In immunohistochemical studies , P25116 -like IR-positive cells in the spinal dorsal horn were mostly colocated on PDGF-like IR-positive neuronal cells . These data provide novel evidence that P25116 and PDGF-A-mediated signaling pathway within spinal cord neurons may be directly implicated in neuropathic pain after nerve injury in mice .

MH_train_1187

MH_train_1187

MH_train_1187

interacts_with DB09026?

multiple_choice

Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . 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 . 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 . [ O6-benzylguanine stimulates regulatory functions of the Ada protein in Escherichia coli ] . In vitro experiments showed that O6-benzylguanine ( O6-benzG , 0.2 microM ) fully inhibited the repair activity of human O6-alkylguanine-DNA alkyltransferase ( P16455 ) due to the formation of S-benzylcytosine in the protein acceptor site . O6-benzG at concentrations increased many times ( up to 800 microM ) failed to inhibit the repair activity of the Escherichia coli Ada protein , the structural and functional analog of P16455 . It has been shown for the first time that O6-benzG stimulates the regulatory activity of the Ada protein . In experiments with N-nitroso-N-methylurea ( P48645 ) , the pretreatment of Escherichia coli cells with O6-benzG at a sublethal concentration of 10 microM led to a twofold enhancement of transcription at the Ada-dependent promoter of the alkA gene in control cells and ensured transcription enhanced 1.6-1.7 times at alkA and alkB promoters in cells with the induced " classical " Ada response . Apparently , an increase in the regulatory activity of the Ada protein was associated with the formation of the stable protein molecule having the strong affinity for alkA and alkB promoters after transfer of the benzyl group from O6-benzG to the acceptor site DB00151 -69 in the N-terminal domain of Ada protein . O6-benzG did not affect the regulative activity of Ada in alternative quasi-adaptive responses to P48645 . 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 . Review : molecular pathology in adult high-grade gliomas : from molecular diagnostics to target therapies . The classification of malignant gliomas is moving from a morphology-based guide to a system built on molecular criteria . The development of a genomic landscape for gliomas and a better understanding of its functional consequences have led to the development of internally consistent molecular classifiers . However , development of a biologically insightful classification to guide therapy is still a work in progress . Response to targeted treatments is based not only on the presence of drugable targets , but rather on the molecular circuitry of the cells . Further , tumours are heterogeneous and change and adapt in response to drugs . Therefore , the challenge of developing molecular classifiers that provide meaningful ways to stratify patients for therapy remains a major challenge for the field . In this review , we examine the potential role of P16455 methylation , O75874 /2 mutations , 1p/19q deletions , aberrant epidermal growth factor receptor and PI3K pathways , abnormal p53/Rb pathways , cancer stem-cell markers and microRNAs as prognostic and predictive molecular markers in the setting of adult high-grade gliomas and we outline the clinically relevant subtypes of glioblastoma with genomic , transcriptomic and proteomic integrated analyses . Furthermore , we describe how these advances , especially in epidermal growth factor receptor/PI3K/ P42345 signalling pathway , affect our approaches towards targeted therapy , raising new challenges and identifying new leads . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling . 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 . Ventilation-induced increases in P00533 ligand mRNA are not altered by intra-amniotic LPS or ureaplasma in preterm lambs . Chorioamnionitis and mechanical ventilation are associated with bronchopulmonary dysplasia ( BPD ) in preterm infants . Mechanical ventilation at birth activates both inflammatory and acute phase responses . These responses can be partially modulated by previous exposure to intra-amniotic ( IA ) LPS or Ureaplasma parvum ( UP ) . P00533 ( P00533 ) ligands participate in lung development , and angiotensin converting enzyme ( P12821 ) 1 and Q9BYF1 contribute to lung inflammation . We asked whether brief mechanical ventilation at birth altered P00533 and P12821 pathways and if antenatal exposure to IA LPS or UP could modulate these effects . Ewes were exposed to IA injections of UP , LPS or saline multiple days prior to preterm delivery at 85 % gestation . Lambs were either immediately euthanized or mechanically ventilated for 2 to 3 hr . IA UP and LPS cause modest changes in the P00533 ligands amphiregulin ( P15514 ) , epiregulin ( O14944 ) , heparin binding epidermal growth factor ( HB- P01133 ) , and betacellulin ( P35070 ) mRNA expression . Mechanical ventilation greatly increased mRNA expression of P15514 , O14944 , and HB- P01133 , with no additional increases resulting from IA LPS or UP . With ventilation P15514 and O14944 mRNA localized to cells in terminal airspace . P00533 mRNA also increased with mechanical ventilation . IA UP and LPS decreased ACE1 mRNA and increased Q9BYF1 mRNA , resulting in a 4 fold change in the ACE1/ Q9BYF1 ratio . Mechanical ventilation with large tidal volumes increased both ACE1 and Q9BYF1 expression . The alterations seen in P12821 with IA exposures and P00533 pathways with mechanical ventilation may contribute to the development of BPD in preterm infants . 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 . A randomized , placebo-controlled study of the effects of the p38 MAPK inhibitor SB- DB05250 on blood biomarkers of inflammation in P48444 patients . The p38 mitogen-activated protein kinase ( MAPK ) signaling upregulates inflammation and is known to be increased in chronic obstructive pulmonary disease ( P48444 ) . The authors assessed the pharmacology of the novel p38 MAPK inhibitor SB- DB05250 using blood biomarkers in P48444 . Seventeen P48444 patients ( forced expiratory volume in 1 second 50 % -80 % predicted ) using short-acting bronchodilators participated in a double-blind , double-dummy , randomized , crossover study . Patients received single oral doses of SB- DB05250 7.5 mg and 25 mg , prednisolone 10 mg and 30 mg , and placebo . Blood was obtained predose and at 1 , 2 , 6 , and 24 hours postdose . Whole-blood sorbitol-induced phosphorylated ( p ) heat shock protein ( HSP ) 27 levels as a marker of p38 pathway activation and lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -alpha production were assessed . Both doses of SB- DB05250 , but not prednisolone , significantly ( P < .0001 ) reduced weighted mean ( WM ) pHSP27 ( 0-6 hours ) by 58 % compared with placebo . WM P01375 production ( 0-24 hours ) was significantly reduced compared with placebo by SB- DB05250 25 mg ( 40 % , P = .005 ) and 7.5 mg ( 33.4 % , P = .02 ) , while prednisolone 30 mg and 10 mg caused 81.5 % and 58.2 % suppression , respectively ( both P < .0001 ) . SB- DB05250 inhibited the p38 MAPK pathway to a greater degree than prednisolone did . SB- DB05250 inhibited P01375 production . SB- DB05250 is a potent p38 MAPK inhibitor that potentially suppresses inflammation in P48444 . Inhibition of toll-like receptor 2 expression improves insulin sensitivity and signaling in muscle and white adipose tissue of mice fed a high-fat diet . The aims of the present study were to investigate the expression of toll-like receptor 2 ( O60603 ) in muscle and white adipose tissue ( WAT ) of diet-induced obesity ( DIO ) mice , and also the effects of its inhibition , with the use of O60603 antisense oligonucleotide ( ASON ) , on insulin sensitivity and signaling . The expression of O60603 was increased in muscle and WAT of DIO mice , compared with those that received standard chow . Inhibition of O60603 in DIO mice , by O60603 ASON , improved insulin sensitivity and signaling in muscle and WAT . In addition , data show that the inhibition of O60603 expression prevents the activation of O14920 , P45983 , and serine phosphorylation of P35568 in DIO mice , suggesting that O60603 is a key modulator of the crosstalk between inflammatory and metabolic pathways . We , therefore , suggest that a selective interference with O60603 presents an attractive opportunity for the treatment of insulin resistance in obesity and type 2 diabetes . The in vitro pharmacological profile of DB05655 , a selective 5-HT(4) receptor agonist with high intrinsic activity . The in vitro pharmacological profile of DB05655 , a novel , selective 5-HT(4) receptor agonist , was compared to that of clinically efficacious gastroprokinetic 5-HT(4) receptor agonists . DB05655 produced an elevation of cyclic adenosine monophosphate in human embryonic kidney 293 cells expressing the human recombinant 5-HT(4(c)) ( h5-HT(4(c)) ) receptor ( pEC(50) = 8.3 ) and 5-HT(4) receptor-mediated relaxation of the rat esophagus ( pEC(50) = 7.9 ) and contraction of the guinea pig colon ( pEC(50) = 7.9 ) . In all in vitro assays , DB05655 was a high intrinsic activity agonist , unlike tegaserod , mosapride , and cisapride which , in the majority of test systems , had lower intrinsic activity . DB05655 had high affinity ( pK ( i ) = 7.7 ) and selectivity ( > or =25-fold ) for h5-HT(4(c)) receptors over other biogenic amine receptors . DB05655 was > 500-fold selective over other 5-HT receptors ( including h5-HT(2B) and h5-HT(3A) ) and , at 3 microM , had no effect on human ether-à-go-go-related gene K+ channels . In conclusion , DB05655 is a selective 5-HT(4) receptor agonist in vitro . The high intrinsic activity and preferential binding of DB05655 to Q13639 over other 5-HT receptors may result in an improved clinical profile for the treatment of gastrointestinal disorders of reduced motility . Local renin-angiotensin II systems , angiotensin-converting enzyme and its homologue Q9BYF1 : their potential role in the pathogenesis of chronic obstructive pulmonary diseases , pulmonary hypertension and acute respiratory distress syndrome . P00797 -angiotensin II-aldosterone axis has long been known as a regulator of blood pressure and fluid homeostasis . Yet , local renin-angiotensin II systems have been discovered and novel actions of angiotensin II ( AngII ) have emerged among which its ability to act as a immunomodulator and profibrotic molecule . The enzyme responsible for its synthesis , Angiotensin-converting-enzyme ( P12821 ) , is present in high concentrations in lung tissue . In the present paper , we review data from studies of the past decade that implicate AngII and functional polymorphisms of the P12821 gene that increase P12821 activity with increased susceptibility for asthma and chronic obstructive pulmonary disease ( P48444 ) and for pulmonary hypertension . Moreover , drugs that inhibit the synthesis of AngII ( P12821 inhibitors ) or that antagonize its actions on its receptors ( Angiotensin II receptor blockers -ARBs ) have been shown to provide beneficial effects . Another recent discovery reviewed is the presence of a homologue of P12821 , Q9BYF1 , which cleaves a single amino acid from AngII and forms a heptapeptide with vasodilatory actions , Ang 1-7 . The balance between P12821 and Q9BYF1 is crucial for controlling AngII levels . P12821 and Q9BYF1 also appear to modify the severity of Acute Respiratory Distress Syndrome ( ARDS ) , with Q9BYF1 playing a protective role . Finally , mention is made to the recent discovery of Q9BYF1 as a receptor for the P49591 Corona Virus . Interactome mapping of the phosphatidylinositol 3-kinase-mammalian target of rapamycin pathway identifies deformed epidermal autoregulatory factor-1 as a new glycogen synthase kinase-3 interactor . The phosphatidylinositol 3-kinase-mammalian target of rapamycin ( PI3K- P42345 ) pathway plays pivotal roles in cell survival , growth , and proliferation downstream of growth factors . Its perturbations are associated with cancer progression , type 2 diabetes , and neurological disorders . To better understand the mechanisms of action and regulation of this pathway , we initiated a large scale yeast two-hybrid screen for 33 components of the PI3K- P42345 pathway . Identification of 67 new interactions was followed by validation by co-affinity purification and exhaustive literature curation of existing information . We provide a nearly complete , functionally annotated interactome of 802 interactions for the PI3K- P42345 pathway . Our screen revealed a predominant place for glycogen synthase kinase-3 ( GSK3 ) A and B and the AMP-activated protein kinase . In particular , we identified the deformed epidermal autoregulatory factor-1 ( O75398 ) transcription factor as an interactor and in vitro substrate of P49840 and P49841 . Moreover , GSK3 inhibitors increased O75398 transcriptional activity on the P08908 serotonin receptor promoter . We propose that O75398 may represent a therapeutic target of lithium and other GSK3 inhibitors used in bipolar disease and depression . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . P05019 attenuates FFA-induced activation of P45983 phosphorylation and TNFα expression in human subcutaneous preadipocytes . OBJECTIVE : Free fatty acids ( FFAs ) are increased in visceral fat and contribute to insulin resistance through multiple mechanisms , including c-Jun N-terminal kinase ( JNK ) activation and expression of TNFα . Given that insulin-like growth factor-1 ( DB01277 ) -mediated proliferation is impaired in omental compared to subcutaneous ( SC ) preadipocytes , we investigated P05019 anti-inflammatory action in preadipocytes from SC and omental adipose tissue . DESIGN AND METHODS : Preadipocytes isolated from abdominal SC and omental fat of obese subjects were studied in primary culture . Cells were exposed to FFAs with or without P05019 pretreatment followed by analysis of cytokine expression and JNK phosphorylation . Lentivirus infection was used to express a constitutively active AKT ( myr-AKT ) in omental preadipocytes . RESULTS : FFAs increased the expression of tumor necrosis factor ( P01375 )α , interleukin ( IL ) -6 , and monocyte chemotactic protein ( MCP ) -1 in SC and omental preadipocytes . P05019 pretreatment reduced FFA-induced P45983 phosphorylation and TNFα expression in SC but not omental preadipocytes . Treatment with the P45983 /2 inhibitor SP600125 reduced FFA-induced expression of TNFα . FFAs and MALP-2 , a specific O60603 /6 ligand , but not specific ligands for O00206 and Q15399 /2 , increased P45983 phosphorylation . P05019 completely inhibited MALP-2-stimulated phosphorylation of P45983 . Expression of myr-AKT in omental preadipocytes inhibited FFA-stimulated P45983 phosphorylation . CONCLUSIONS : P05019 attenuated FFA-induced P45983 phosphorylation and TNFα expression through activation of AKT in human subcutaneous but not omental preadipocytes . Effects of lesopitron on the central nervous system arising from its interaction with P08908 receptors . DB04970 acts as a ligand for central serotonin P08908 receptors . Ki obtained from [3H]8-OH-DPAT competition studies was 104.8 +/- 10.6 nmol/l . As lesopitron did not affect the binding of [3H]paroxetine , involvement of the serotonin reuptake system in the effects of lesopitron is rejected . DB04970 inhibits haloperidol-induced catalepsy that is the consequence of its action on P08908 autoreceptors . The ability of lesopitron to induce 5-HT syndrome reflects post-synaptic P08908 receptor activation and the reversion of 8-OHDPAT-induced 5-HT syndrome by lesopitron suggests a partial agonist effect on this receptor-type . DB04970 induced a hypothermic effect due to the enhanced activation of post-synaptic P08908 receptors . The agonist effect of lesopitron on P08908 receptors and its marked hypothermic effect is an added value for this drug and a stimulus to the study of its possible neuroprotective action . Investigation of Q13639 receptors in bronchial hyperresponsiveness in cigarette smoke-exposed mice . BACKGROUND : Chronic obstructive pulmonary disease ( P48444 ) arises from an interaction between genetic host factors and environmental exposures ( mainly cigarette smoke ( CS ) ) . Genome Wide Association studies have demonstrated that genetic variations in the gene encoding 5-hydroxytryptamine 4 receptors ( 5-HT(4)R ) , Q13639 , were associated with measures of airway obstruction and with P48444 . We hypothesised that 5-HT(4) receptors , in addition to 5-HT2AR and muscarinic receptors , contribute to the pathogenesis of P48444 by facilitating cholinergic bronchoconstriction . METHODS : The levels of pulmonary 5-HT(4)R mRNA were measured in CS-exposed mice by qRT-PCR . We investigated the effect of CS exposure on bronchial hyperresponsiveness ( BHR ) to 5-HT and evaluated the contribution of 5-HT2AR , muscarinic receptors and 5-HT(4)R in the response to 5-HT by using the corresponding antagonists and 5-HT(4)R knockout ( KO ) mice . RESULTS : The 5-HT(4)R mRNA levels were significantly elevated upon acute ( 3 days ) , subacute ( 4 weeks ) and chronic ( 24 weeks ) CS exposure . Both acute and subacute CS exposure significantly increased BHR to 5-HT . Antagonism of 5-HT2AR abolished the CS-induced BHR to 5-HT , and antagonism of muscarinic receptors significantly reduced the response to 5-HT . However , pre-treatment with GR113808 , a specific 5-HT(4)R antagonist , did not alter the response to 5-HT in CS-exposed mice . Accordingly , the CS-induced BHR to 5-HT was not different between wild-type and 5-HT(4)R KO mice . CONCLUSION : CS increased the levels of 5-HT(4)R mRNA in the lungs , concomitantly with bronchial responsiveness to 5-HT . Our in vivo data using pharmacologic and genetic approaches suggest that 5-HT(4) receptors are not involved in the BHR to 5-HT in CS-exposed mice . 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 . Evidence for epidermal growth factor receptor as negative-feedback control in aldosterone-induced Na+ reabsorption . DB04630 enhances Na(+) reabsorption via epithelial Na(+) channels ( ENaC ) . DB04630 also stimulates the protein kinase P27361 /2- and the epidermal growth factor ( P01133 ) receptor ( P00533 ) -signaling pathway . Yet P01133 and P27361 /2 are known inhibitors of ENaC-mediated Na(+) reabsorption . In the present study , using the well-established Madin-Darby canine kidney P10643 cell line , we tested the hypothesis that P00533 represents a negative-feedback control for chronic aldosterone-induced Na(+) reabsorption [ amiloride-inhibitable short-circuit current ( I(sc) ) ] . P08235 expression was confirmed by RT-PCR and Western blot analysis . DB04630 enhanced P27361 /2 phosphorylation in an P00533 -dependent way . Furthermore , aldosterone stimulated P00533 expression . DB04630 ( 10 nmol/l ) induced a small transient increase in I(sc) under control conditions . Inhibition of P27361 /2 phosphorylation with U-0126 ( 10 micromol/l ) stimulated I(sc) , indicating constitutive ENaC inhibition . DB04630 exerted a significantly larger effect in the presence of U-0126 than without U-0126 . P01133 ( 10 microg/l ) inhibited I(sc) , whereas inhibition of P00533 kinase by tyrphostin AG-1478 ( 100 nmol/l ) enhanced I(sc) . DB04630 was more effective in the presence of AG-1478 than without AG-1478 . In summary , we propose that the P00533 -signaling cascade can serve as a negative-feedback control to limit the effect of aldosterone-induced Na(+) reabsorption .

MH_train_1188

MH_train_1188

MH_train_1188

interacts_with DB00452?

multiple_choice

Modeling the neurovascular niche : murine strain differences mimic the range of responses to chronic hypoxia in the premature newborn . Preterm birth results in significant cognitive and motor disabilities , but recent evidence suggests that there is variable recovery over time . One possibility that may explain this variable recovery entails variable neurogenic responses in the subventricular zone ( SVZ ) following the period of chronic hypoxia experienced by these neonates . In this report , we have characterized the responses to chronic hypoxia of two mouse strains that represent a wide range of susceptibility to chronic hypoxia . We determined that C57BL/6 pups and neural progenitor cells ( NPCs ) derived from them exhibit a blunted response to hypoxic insult compared with CD-1 pups and NPCs . Specifically , C57BL/6 pups and NPCs exhibited blunted in vivo and in vitro proliferative and increased apoptotic responses to hypoxic insult . Additionally , C57BL/6 NPCs exhibited lower baseline levels and hypoxia-induced levels of selected transcription factors , growth factors , and receptors ( including HIF-1alpha , Q9GZT9 , P23560 , P15692 , P48061 , TrkB , Nrp-1 , P61073 , and NO ) that determine , in part , the responsiveness to chronic hypoxic insult compared with CD-1 pups and NPCs , providing insight into this important and timely problem in perinatology . Inhibition of chemokine ( CXC motif ) ligand 12/chemokine ( CXC motif ) receptor 4 axis ( P48061 / P61073 ) -mediated cell migration by targeting mammalian target of rapamycin ( P42345 ) pathway in human gastric carcinoma cells . P48061 / P61073 plays an important role in metastasis of gastric carcinoma . DB00877 has been reported to inhibit migration of gastric cancer cells . However , the role of P42345 pathway in P48061 / P61073 -mediated cell migration and the potential of drugs targeting PI3K/ P42345 pathway remains unelucidated . We found that P48061 activated PI3K/Akt/ P42345 pathway in MKN-45 cells . Stimulating CHO- P04264 cells expressing pEGFP-C1- O43739 -PH fusion protein with P48061 resulted in generation of phosphatidylinositol (3,4,5)-triphosphate , which provided direct evidence of activating PI3K by P48061 . Down-regulation of p110β by siRNA but not p110α blocked phosphorylation of Akt and P23443 induced by P48061 . Consistently , p110β-specific inhibitor blocked the P48061 -activated PI3K/Akt/ P42345 pathway . Moreover , P61073 immunoprecipitated by anti-p110β antibody increased after P48061 stimulation and G(i) protein inhibitor pertussis toxin abrogated P48061 -induced activation of PI3K . Further studies demonstrated that inhibitors targeting the PI3K/ P42345 pathway significantly blocked the chemotactic responses of MKN-45 cells triggered by P48061 , which might be attributed primarily to inhibition of mTORC1 and related to prevention of F-actin reorganization as well as down-regulation of active RhoA , Rac1 , and Cdc42 . Furthermore , rapamycin inhibited the secretion of P48061 and the expression of P61073 , which might form a positive feedback loop to further abolish upstream signaling leading to cell migration . Finally , we found cells expressing high levels of cxcl12 were sensitive to rapamycin in its activity inhibiting migration as well as proliferation . In summary , we found that the P42345 pathway played an important role in P48061 / P61073 -mediated cell migration and proposed that drugs targeting the P42345 pathway may be used for the therapy of metastatic gastric cancer expressing high levels of cxcl12 . High-density lipoproteins prevent the oxidized low-density lipoprotein-induced epidermal [ corrected ] growth factor receptor activation and subsequent matrix metalloproteinase-2 upregulation . OBJECTIVE : The atherogenic oxidized low-density lipoprotein ( oxLDL ) induces the formation of carbonyl-protein adducts and activates the epidermal [ corrected ] growth factor receptor ( P00533 ) signaling pathway , which is now regarded as a central element for signal transduction . We aimed to investigate whether and by which mechanism the anti-atherogenic high-density lipoprotein ( HDL ) prevents these effects of oxLDL . METHODS AND RESULTS : In vascular cultured cells , HDL and apolipoprotein A-I inhibit oxLDL-induced P00533 activation and subsequent signaling by acting through 2 separate mechanisms . First , HDL , like the aldehyde scavenger dinitrophenyl hydrazine , prevented the formation of oxLDL-induced carbonyl-protein adducts and 4-hydroxynonenal ( HNE ) - P00533 adducts . Secondly , HDL enhanced the cellular antioxidant defenses by preventing ( through a scavenger receptor class B-1 ( Q8WTV0 ) -dependent mechanism ) the increase of intracellular reactive oxygen species ( ROS ) and subsequent P00533 activation triggered by oxLDL or H2O2 . A pharmacological approach suggests that this protective effect of HDL is independent of cellular glutathione level and glutathione peroxidase activity , but it requires catalase activity . Finally , we report that oxLDL upregulates both membrane type 1 ( MT1 ) -matrix metalloproteinase-1 ( P50281 ) and P08253 through an P00533 -dependent mechanism and that HDL inhibits these events . CONCLUSIONS : HDLs block in vitro oxLDL-induced P00533 signaling and subsequent P08253 activation by inhibiting carbonyl adducts formation and cellular oxidative stress . These effects of HDL may participate to reduce cell activation , excessive remodeling , and alteration of the vascular wall . Signal transduction pathways of the human V1-vascular , V2-renal , V3-pituitary vasopressin and oxytocin receptors . DB00067 ( VP ) and oxytocin ( OT ) are cyclic nonapeptides whose actions are mediated by stimulation of specific G protein-coupled receptors ( GPCRs ) currently classified into V1-vascular ( V1R ) , V2-renal ( P30518 ) and V3-pituitary ( V3R ) VP receptors and OT receptors ( OTR ) . The recent cloning of the different members of the VP/OT family of receptors now allows the extensive characterization of the molecular determinants involved in ligand binding and signal transduction pathways coupled to a given VP/OT receptor subtype in stably transfected mammalian cell lines . In this article , we review the present knowledge of the signal transduction pathways coupled to the different VP/OT receptor subtypes and we present new observations derived from the study of each human VP or OT receptor subtype stably expressed in CHO cells . DB00144 binding of class B scavenger receptor type I , a phagocytosis receptor of testicular sertoli cells . Testicular Sertoli cells phagocytose apoptotic spermatogenic cells in a manner depending on the membrane phospholipid phosphatidylserine ( PS ) expressed at the surface of the latter cell type . Our previous studies have indicated that class B scavenger receptor type I ( Q8WTV0 ) is responsible for the PS-mediated phagocytosis by Sertoli cells . We examined here whether Q8WTV0 binds directly to PS . A cell line acquired the ability to bind to PS-exposing apoptotic cells and to incorporate PS-containing liposomes when it was forced to express Q8WTV0 . Furthermore , the extracellular domain of rat Q8WTV0 fused with human Fc ( SRBIecd-Fc ) bound to PS with a dissociation equilibrium constant of 2.4 x 10(-7) m in a cell-free solid-phase assay , whereas other phospholipids including phosphatidylethanolamine , phosphatidylinositol , and phosphatidylcholine were poor binding targets . The binding activity was enhanced when CaCl(2) was included in the assay or when SRBIecd-Fc was pre-treated with N-glycanase . A portion of the extracellular domain spanning amino acid positions 33 and 191 ( numbered with respect to the amino terminus ) fused with Fc ( SRBI33-191-Fc ) showed activity and phospholipid specificity equivalent to those of SRBIecd-Fc . Finally , SRBI33-191-Fc bound to the surface of apoptotic cells with externalized PS , and the injection of SRBI33-191-Fc into the seminiferous tubules of live mice increased the number of apoptotic spermatogenic cells . These results allowed us to conclude that Q8WTV0 is a phagocytosis-inducing PS receptor of Sertoli cells . 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 . 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 . Intracellular retention and degradation of the epidermal growth factor receptor , two distinct processes mediated by benzoquinone ansamycins . Epidermal growth factor ( P01133 ) stimulates the growth of various types of cells via its cell surface tyrosine kinase receptor . The P01133 receptor ( P01133 -R ) has an oncogenic potential when overexpressed in a wide range of tumor cells . DB02424 ( GA ) and herbimycin ( HA ) , specific inhibitors of the cytosolic chaperone P08238 and its endoplasmic reticulum homologue GRP 94 , were shown to accelerate degradation of the P01133 -R and of its homologue p185(c-)(erbB-2) . Here we compared the effects of GA and HA on intracellular degradation and maturation of P01133 -R . By using an inhibitor of proteasomal degradation , we learned that GA , but not HA , blocks processing of newly synthesized P01133 -R . The effects of GA and HA on receptor degradation are mediated by the cytosolic portion of P01133 -R and could be conferred to the erythropoietin receptor ( P19235 ) , by employing the respective chimera . Neither HA nor GA affected stability of newly synthesized P01133 -R lacking the cytosolic domain ( Ex P01133 -R ) , but GA caused intracellular retention of this mutant . Taken together , our results imply that GA has two distinct targets of action on the P01133 -R , one for promoting its degradation and another for mediating its intracellular retention . Apparently , degradation of the P01133 -R mediated by GA or HA requires the presence of the P01133 -R cytosolic domain , whereas intracellular retention in the presence of GA is coupled to the extracellular domain of the P01133 -R . Highly metastatic hepatocellular carcinomas induced in male F344 rats treated with N-nitrosomorpholine in combination with other hepatocarcinogens show a high incidence of p53 gene mutations along with altered mRNA expression of tumor-related genes . The carcinogenic and metastatic processes are thought to consist of a sequence of steps , and animal models featuring highly metastatic lesions are clearly necessary to allow analysis of the whole process of transformation from preneoplastic changes to high grade metastatic tumors , and to access effectiveness of therapeutic treatments of advanced cancers in vivo . The purpose of the present study was to establish a model and to screen for reported genetic alterations in induced lesions . In the present study , it was confirmed that lung metastasis of hepatocellular carcinomas ( HCCs ) induced in male F344 rats by N-nitrosomorpholine ( NNM ) , given in the drinking water at a dose of 120 ppm for 24 weeks , was significantly enhanced by additional carcinogenic pretreatments and that a single i.p. injection of 100 mg/kg body weight N-diethylnitrosamine ( DEN ) alone was sufficient for that purpose . Molecular biological analyses of the induced lesions revealed point mutations in the p53 gene in 60.9 % of HCCs , and elevated expression of mRNAs for p53 , c-myc , c-fos , TGF-alpha , TGF-beta1 , alpha-fetoprotein , Q86UG4 -P , and P19440 , and decreased mRNA expression of P01133 and P00533 in HCCs when compared to controls . No obvious association of gene alterations with metastatic potential of primary tumors was found except for an increase in the incidence of p53 mutations . Since the process of metastasis is thought to be sequential and selective , further comparative analysis of metastatic and primary lesions should clarify the mechanisms involved in the multi-step process of metastasis . The cleaved cytoplasmic tail of polycystin-1 regulates Src-dependent P40763 activation . P98161 ( PC1 ) mutations result in proliferative renal cyst growth and progression to renal failure in autosomal dominant polycystic kidney disease ( ADPKD ) . The transcription factor P40763 ( signal transducer and activator of transcription 3 ) was shown to be activated in cyst-lining cells in ADPKD and Q15139 mouse models and may drive renal cyst growth , but the mechanisms leading to persistent P40763 activation are unknown . A proteolytic fragment of PC1 corresponding to the cytoplasmic tail , PC1-p30 , is overexpressed in ADPKD . Here , we show that PC1-p30 interacts with the nonreceptor tyrosine kinase Src , resulting in Src-dependent activation of P40763 by tyrosine phosphorylation . The PC1-p30-mediated activation of Src/ P40763 was independent of JAK family kinases and insensitive to the P40763 inhibitor suppressor of cytokine signaling 3 . Signaling by the P01133 receptor ( P00533 ) or DB02527 amplified the activation of Src/ P40763 by PC1-p30 . Expression of PC1-p30 changed the cellular response to DB02527 signaling . In the absence of PC1-p30 , DB02527 dampened P00533 - or P05231 -dependent activation of P40763 ; in the presence of PC1-p30 , DB02527 amplified Src-dependent activation of P40763 . In the polycystic kidney ( PCK ) rat model , activation of P40763 in renal cystic cells depended on vasopressin receptor 2 ( P30518 ) signaling , which increased DB02527 levels . Genetic inhibition of vasopressin expression or treatment with a pharmacologic P30518 inhibitor strongly suppressed P40763 activation and reduced renal cyst growth . These results suggest that PC1 , via its cleaved cytoplasmic tail , integrates signaling inputs from P00533 and DB02527 , resulting in Src-dependent activation of P40763 and a proliferative response . Comparative effects of azimilide and ambasilide on the human ether-a-go-go-related gene ( Q12809 ) potassium channel . OBJECTIVE : To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related ( Q12809 ) channel . METHODS : Q12809 was stably transfected into Chinese hamster ovary ( CHO- P04264 ) cells and currents were measured using a whole cell , voltage-clamp technique . RESULTS : DB04957 had a ' dual effect ' , inhibiting current at voltage steps above -40 mV and augmenting current at -40 and -50 mV . Tail current inhibition following a step to +30 mV did not vary with temperature ( IC(50) 610 nM at 22 degrees C and 560 nM at 37 degrees C ) . The agonist effect at -50 mV was concentration-dependent and correlated with a hyperpolarizing shift in the V(1/2) of activation ( r=0.98 , P < 0.05 ) . Time constants of inactivation were faster and there was a -10 mV shift in the V(1/2) of steady state inactivation suggestive of open and inactivated state binding . By comparison , ambasilide inhibited Q12809 channels with lower potency ( IC(50) 3.6 microM ) , in a voltage- and time-dependent but frequency-independent manner ( 0.03-1 Hz ) . Ambasilide had no effect on activation or inactivation gating but prolonged both fast and slow components of deactivation consistent with unbinding from the open state . The net effect of both drugs was similar during a voltage ramp which simulated a cardiac action potential . CONCLUSIONS : Inhibition of Q12809 channels by azimilide and ambasilide exhibits a similar time and voltage-dependence . While both exhibit affinity for the open state , azimilide also binds to inactivated channels . DB04873 ( SB 207266 ) , a selective Q13639 receptor antagonist , reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle . The aim of this study is to evaluate the potency of piboserod ( SB 207266 ) , a selective 5-HT(4) receptor antagonist , at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin ( 5-HT ) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations ( O43281 ) . Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and O43281 ( 20 Hz , 1 ms duration at 300 mA for 5 s ) was applied continuously at 1 min intervals . After stabilization of the O43281 -induced contractions , concentration-response curves to 5-HT ( 0.1 nM-100 microM ) were constructed in the absence or presence of 1 or 100 nM of piboserod . The experiments were performed in the presence of methysergide ( 1 microM ) and ondansetron ( 3 microM ) to block 5HT(1)/5HT(2) and 5-HT(3) receptors , respectively . 5-HT potentiated the contractile responses to O43281 of human bladder strips in a concentration-dependent manner , with a maximum mean of 60.0+/-19.9 % of the basal O43281 -evoked contractions . DB04873 did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to O43281 . In presence of 1 and 100 nM of piboserod , the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7 % , respectively . A mean apparent antagonist dissociation constant value ( K(B) ) of 0.56+/-0.09 nM was determined . These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally-mediated contractions of isolated human bladder strips . Therefore , the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder . Autocrine stimulation of P35968 activates human leukemic cell growth and migration . Emerging data suggest that P15692 receptors are expressed by endothelial cells as well as hematopoietic stem cells . Therefore , we hypothesized that functional P15692 receptors may also be expressed in malignant counterparts of hematopoietic stem cells such as leukemias . We demonstrate that certain leukemias not only produce P15692 but also express functional P35968 in vivo and in vitro , resulting in the generation of an autocrine loop that may support leukemic cell survival and proliferation . Approximately 50 % of freshly isolated leukemias expressed mRNA and protein for P35968 . P15692 (165) induced phosphorylation of P35968 and increased proliferation of leukemic cells , demonstrating these receptors were functional . P15692 (165) also induced the expression of P14780 by leukemic cells and promoted their migration through reconstituted basement membrane . The neutralizing mAb DB06101 , specific to human P35968 , inhibited leukemic cell survival in vitro and blocked P15692 (165)-mediated proliferation of leukemic cells and P15692 -induced leukemic cell migration . Xenotransplantation of primary leukemias and leukemic cell lines into immunocompromised nonobese diabetic mice resulted in significant elevation of human , but not murine , P15692 in plasma and death of inoculated mice within 3 weeks . Injection of DB06101 inhibited proliferation of xenotransplanted human leukemias and significantly increased the survival of inoculated mice . Interruption of signaling by VEGFRs , particularly P35968 , may provide a novel strategy for inhibiting leukemic cell proliferation . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . DB00160 aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis . Plant peroxisomal glyoxylate aminotransferases play central roles within the photorespiratory pathway . Genes encoding glyoxylate aminotransferases have been isolated from several animals and microbes , but only recently have plant homologs been identified . Three Arabidopsis homologs of alanine (Ala):glyoxylate aminotransferase 2 ( Q9BYV1 ) contain a putative type 1 peroxisomal targeting signal ( PTS1 ) , but the metabolic significance of these Q9BYV1 homologs is unknown . P19440 and P36268 are Ala aminotransferase ( AlaAT ) homologs from Arabidopsis that represent another type of glyoxylate aminotransferase . These proteins are class I aminotransferases , each containing a putative PTS1 . P19440 and P36268 are members of a small family of AlaATs in Arabidopsis . When expressed as recombinant proteins in Escherichia coli , P19440 and P36268 displayed biochemical characteristics very similar to one another , and to the Arabidopsis protein purified from leaves . Four aminotransferase activities were specifically associated with P19440 and P36268 , using the substrate pairs glutamate ( DB00142 ):glyoxylate , Ala:glyoxylate , DB00142 :pyruvate , and Ala:2-oxoglutarate . P19440 and P36268 may have partially redundant functions ; transcripts of both genes were detected in many of the same tissues . Although DB00142 :glyoxylate aminotransferase ( P19440 ) activity has been observed in several locations in different plants and algae , including the cytoplasm and mitochondria , our subcellular fractionation data indicate that P19440 activity was exclusively peroxisomal in Arabidopsis . Thus , glyoxylate aminotransferase reactions in plant peroxisomes appear to be catalyzed by at least two distinct types of aminotransferases : an AGT1 homolog with serine:glyoxylate aminotransferase activity ( A.H. Liepman , L.J. Olsen [ 2001 ] Plant J 25 : 487-498 ) , and a pair of closely related , potentially redundant AlaAT homologs with P19440 activity . 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 . 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 . A whole genome Bayesian scan for adaptive genetic divergence in West African cattle . BACKGROUND : The recent settlement of cattle in West Africa after several waves of migration from remote centres of domestication has imposed dramatic changes in their environmental conditions , in particular through exposure to new pathogens . West African cattle populations thus represent an appealing model to unravel the genome response to adaptation to tropical conditions . The purpose of this study was to identify footprints of adaptive selection at the whole genome level in a newly collected data set comprising 36,320 SNPs genotyped in 9 West African cattle populations . RESULTS : After a detailed analysis of population structure , we performed a scan for SNP differentiation via a previously proposed Bayesian procedure including extensions to improve the detection of loci under selection . Based on these results we identified 53 genomic regions and 42 strong candidate genes . Their physiological functions were mainly related to immune response ( MHC region which was found under strong balancing selection , P11912 , P61073 , P80370 , P48380 , Q9H3S1 , Q8IUC6 and P19474 ) , nervous system ( Q96NK8 , O95897 , MAGI1 , Q9H3S1 and Q13639 ) and skin and hair properties ( P24530 , TRSP1 and Q8IUC2 ) . CONCLUSION : The main possible underlying selective pressures may be related to climatic conditions but also to the host response to pathogens such as Trypanosoma(sp) . Overall , these results might open the way towards the identification of important variants involved in adaptation to tropical conditions and in particular to resistance to tropical infectious diseases .

MH_train_1189

MH_train_1189

MH_train_1189

interacts_with DB00191?

multiple_choice

P30968 signaling : biased and unbiased . DB00644 is a neuropeptide that acts via Gq coupled G-protein coupled receptors in the pituitary that mediate central control of reproduction . DB00644 receptors ( GnRHR ) and DB00644 ligands are also found in extra-pituitary sites including the CNS as well as reproductive tissues and cancer cells derived from such tissues . Much of the interest in the extra-pituitary receptors stems from the fact that they mediate anti-proliferative and/or pro-apoptotic effects and may therefore be directly targeted for cancer therapy . Type I mammalian GnRHR are atypical in that they do not bind to ( or signal via ) arrestins . In spite of this restriction on their signaling repertoire , there is good evidence for existence of multiple active GnRHR conformations and for activation of multiple upstream effectors ( heterotrimeric and monomeric G-proteins ) . In this review GnRHR signaling is described , with emphasis on the relevance of functional selectivity for pharmacological characterization of GnRHR ligands , as well as its possible contribution to contextdependent GnRHR signaling and relevance for GnRHR-mediated effects on cell fate as well as GnRHR trafficking . Expression of serotonergic system components during early Xenopus embryogenesis . Despite abundant research studies on the physiological and biochemical nature of embryonic neurotransmitter function , little is known about the molecular genetic mechanisms involved . The expression of the main components of the serotonergic system during early Xenopus embryogenesis was investigated using RT-PCR , real time PCR and in situ hybridization . Transcripts encoding the serotonin receptors P28335 and P34969 , as well as the vesicular monoamine transporter Q05940 , the serotonin transporter ( P31645 ) and the serotonin synthesis enzymes tryptophan hydroxylase ( Q8IWU9 ) and aromatic amino acid decarboxylase ( AAAD ) were found to be expressed during the cleavage division stages , whereas the degradation enzyme monoamine oxidase A ( P21397 ) was absent . The main components of the serotonergic system were found to be expressed during the earliest stages of embryonic development . The embryonic transmitter mechanism , its complexity , and its variability among various species are discussed . 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 . Convergent and divergent cellular responses by ErbB4 isoforms in mammary epithelial cells . Associations of ErbB4 ( Q15303 / Q15303 ) , the fourth member of the P00533 family , with cancer are variable , possibly as a result of structural diversity of this receptor . There are multiple structural isoforms of Q15303 arising by alternative mRNA splicing , and a subset undergo proteolysis that releases membrane-anchored and soluble isoforms that associate with transcription factors and coregulators to modulate transcription . To compare the differential and common signaling activities of full-length ( FL ) and soluble intracellular isoforms of Q15303 , four JM-a isoforms ( FL and soluble intracellular domain ( ICD ) CYT-1 and CYT-2 ) were expressed in isogenic MCF10A cells and their biologic activities were analyzed . Both FL and ICD CYT-2 promoted cell proliferation and invasion , and CYT-1 suppressed cell growth . Transcriptional profiling revealed several new and underexplored Q15303 -regulated transcripts , including : proteases/protease inhibitors ( P08254 and P07093 ) , the YAP/Hippo pathway ( P29279 , O00622 , and P09486 ) , the mevalonate/cholesterol pathway ( P04035 , Q01581 , P01130 , and Q9UBM7 ) , and cytokines ( P10145 , P78556 , and P09341 ) . Many of these transcripts were subsequently validated in a luminal breast cancer cell line that normally expresses Q15303 . Furthermore , ChIP-seq experiments identified O75689 , P02649 , P09486 , P16949 , and Q05195 as novel molecular targets of Q15303 . These findings clarify the diverse biologic activities of Q15303 isoforms , and reveal new and divergent functions . IMPLICATIONS : ErbB4 as a regulator of Hippo and mevalonate pathways provides new insight into milk production and anabolic processes in normal mammary epithelia and cancer . Large-scale association study for structural soundness and leg locomotion traits in the pig . BACKGROUND : Identification and culling of replacement gilts with poor skeletal conformation and feet and leg ( FL ) unsoundness is an approach used to reduce sow culling and mortality rates in breeding stock . Few candidate genes related to soundness traits have been identified in the pig . METHODS : In this study , 2066 commercial females were scored for 17 traits describing body conformation and FL structure , and were used for association analyses . Genotyping of 121 SNPs derived from 95 genes was implemented using Sequenom 's MassARRAY system . RESULTS : Based on the association results from single trait and principal components using mixed linear model analyses and false discovery rate testing , it was observed that P02649 , P34820 , P30988 , P08123 , P20849 , DKFZ , P35555 and VDBP were very highly significantly ( P < 0.001 ) associated with body conformation traits . The genes P09917 , P34820 , P30988 , O00300 , P30559 and Q9UBV4 were very highly significantly ( P < 0.001 ) associated with FL structures , and P02649 , P30988 , P08123 , P30968 , Q14623 , P42898 and Q9UBV4 were highly significantly ( P < 0.01 ) associated with overall leg action . Strong linkage disequilibrium between P30988 and P08123 on SSC9 was detected , and haplotype -ACGACC- was highly significantly ( P < 0.01 ) associated with overall leg action and several important FL soundness traits . CONCLUSION : The present findings provide a comprehensive list of candidate genes for further use in fine mapping and biological functional analyses . Local inhibition of angiogenesis by halofuginone coated silicone materials . Anti-angiogenic therapy is a promising approach for the treatment of increased angiogenesis in certain diseases . We aimed to investigate the local anti-angiogenic effect of silicone implants coated with DB04866 , an angiogenesis inhibitor that inhibits synthesis of collagen-type-I and matrix metalloproteinases . The degree of angiogenesis was observed after implantation of surface modified DB04866 eluting silicone implants into a submuscular pocket in rats over a period of 3 months . Subsequently , key mediators of angiogenesis ( P01137 , P09038 , P02452 , P08253 , P14780 , P15692 and PDGF ) were established by immunohistological staining and RT-PCR and statistically evaluated . In comparison to uncoated silicone implants , DB04866 eluting silicone implants lead to a significant local decrease of angiogenesis . DB04866 eluting hybrid surface silicone implants have a significant local anti-angiogenic effect by down-regulating the expression activity of key mediators of angiogenesis . Modification of alternative messenger RNA splicing of fibroblast growth factor receptors in human cardiac allografts during rejection . Accelerated coronary atherosclerosis in cardiac transplants ( cardiac allograft vasculopathy , Q03135 ) is characterized by coronary intimal hyperplasia . P05230 ( P05230 ) is a potent mitogen for vascular smooth muscle cells and endothelial cells , and its expression is increased in cardiac allografts , suggesting it may play a role in the pathogenesis of Q03135 . The activity of P05230 is dependent on binding to transmembrane receptors . To investigate whether receptors for P05230 are also induced after transplantation , polymerase chain reaction , in situ hybridization , and immunohistochemistry were used to analyze expression of four receptors for P05230 ( P11362 - P22455 ) . Expression of mRNA encoding extracellular immunoglobulin-like domains of P11362 was increased 35-fold in cardiac allografts compared with normal hearts and was predominantly present in cardiac myocytes and vascular structures . Alternatively spliced mRNA that encodes transmembrane forms of P11362 , which contain the signal-transducing tyrosine kinase domains , was induced in allografts during rejection , in infiltrating cells , vascular structures , and myocytes . In vitro experiments showed that differential expression of FGF receptor isoforms was induced by P05230 , and also by P05231 and TGF-beta , which are expressed in cardiac allografts during rejection . The results show that expression of both P05230 and its receptors is altered in cardiac allografts and suggest that these events are important in the pathogenesis of Q03135 . 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 clinical trial with chimeric monoclonal antibody DB05304 and low dose interleukin-2 pulsing scheme for advanced renal cell carcinoma . PURPOSE : DB05304 is a chimeric monoclonal antibody that binds to carbonic anhydrase IX( Q16790 /MN) , which is present on greater than 95 % of RCCs of the clear cell subtype . The suggested working mechanism of DB05304 is by ADCC . Because the number of activated ADCC effector cells can be increased by a low dose interleukin-2 pulsing schedule , a multicenter study was initiated to investigate whether DB05304 combined with LD- P60568 could lead to an improved clinical outcome in patients with progressive RCC . MATERIALS AND METHODS : A total of 35 patients with progressive clear cell RCC received weekly infusions of DB05304 for 11 weeks combined with a daily LD- P60568 regimen . Patients were monitored longitudinally for ADCC capacity . Radiological assessment of metastatic lesions was performed at week 16 and regularly until disease progression . RESULTS : A durable clinical benefit was achieved in 8 of 35 patients ( 23 % ) , including 3 with a partial response and 5 with stabilization at 24 weeks or greater . Mean survival was 22 months . In general treatment was well tolerated with little toxicity . The number of effector cells increased during treatment but lytic capacity per cell did not increase . ADCC and clinical outcome did not appear to correlate . CONCLUSIONS : DB05304 combined with LD- P60568 in patients with metastatic RCC is safe and well tolerated . With a substantial clinical benefit and a median survival of 22 months in patients with metastatic RCC who have progressive disease at study entry combination therapy showed increased overall survival compared to DB05304 monotherapy . Survival was at least similar to that of currently used cytokine regimens but with a favorable toxicity profile . Mechanism of impaired local hyaluronan turnover in bleomycin-induced lung injury in rat . DB08818 , a linear polysaccharide , is accumulated in lung interstitium during different pathological conditions , causing interstitial edema and thereby impaired lung function . We investigated the mechanism of local hyaluronan turnover during the early phase of bleomycin-induced fibrotic lung injury in rats . The binding of [3H]hyaluronan to alveolar macrophages ( AM ) established from bleomycin-treated rats 1 and 5 days after induction of injury was decreased 8- and 15-fold , respectively , compared with that of AM from saline-treated control counterparts , but at day 14 returned almost to the normal level . Data was confirmed by quantitative cytochemistry , using fluorescein-labeled hyaluronan . Analysis of the expression of P16070 , a receptor for hyaluronan , by Western blotting revealed a 30 % increase of P16070 molecules expressed on AM from bleomycin-treated rats at day 5 compared with control rats . In particular a lower molecular mass form of P16070 appeared . No expression of the receptor for hyaluronan-mediated motility ( O75330 ) could be detected . The internalization and degradation of [3H]hyaluronan by AM , obtained from bleomycin-treated rats at days 1 , 5 , and 14 , were decreased about 65 % , 35 % , and 30 % , respectively , compared with AM from the control rats . The AM lysosomal hyaluronidase activity did not differ significantly between bleomycin-treated and control rats . Our results indicate that a decreased hyaluronan binding capacity of AM may account for the impairment of internalization and thereby degradation of excessive hyaluronan during the early phase of fibrotic lung injury . The PEPvIII-KLH ( DB05374 ) vaccine in glioblastoma multiforme patients . Conventional therapies for glioblastoma multiforme ( GBM ) fail to target tumor cells exclusively , resulting in non-specific toxicity . Immune targeting of tumor-specific mutations may allow for more precise eradication of neoplastic cells . P00533 variant III ( EGFRvIII ) is a tumor-specific mutation that is widely expressed in GBM and other neoplasms and its expression enhances tumorigenicity . This in-frame deletion mutation splits a codon , resulting in a novel glycine at the fusion junction producing a tumor-specific epitope target for cellular or humoral immunotherapy . We have previously shown that vaccination with a peptide that spans the EGFRvIII fusion junction ( PEPvIII-KLH/ DB05374 ) is an efficacious immunotherapy in syngeneic murine models . In this review , we summarize our results in GBM patients targeting this mutation in multiple , multi-institutional Phase II immunotherapy trials . These trials demonstrated that a selected population of GBM patients who received vaccines targeting EGFRvIII had an unexpectedly long survival time . Further therapeutic strategies and potential pitfalls of using this approach are discussed . PPARγ and RXR ligands disrupt the inflammatory cross-talk in the hypoxic breast cancer stem cells niche . Cancer stem cells ( CSCs ) are affected by the local micro-environment , the niche , in which inflammatory stimuli and hypoxia act as steering factors . Here , two nuclear receptors ( NRs ) agonists , i.e. pioglitazone ( PGZ ) , a ligand of peroxisome proliferator activated receptor-γ , and 6-OH-11-O-hydroxyphenanthrene ( IIF ) , a ligand of retinoid X receptors , were investigated for their capability to interference with the cross-talk between breast CSCs and the niche compartment . We found that IIF potentiates the ability of PGZ to hamper the mammospheres-forming capability of human breast tumours and MCF7 cancer cells , reducing the expression of CSCs regulatory genes ( Notch3 , P78504 , O43623 , P05231 , P02649 , Hypoxia inducible factor-1α and Q16790 ) . Notably , these effects are not observed in normal-MS obtained from human breast tissue . Importantly , NRs agonists abolish the capability of hypoxic MCF7 derived exosomes to induce a pro-inflammatory phenotype in mammary glands fibroblasts . Moreover , NRs agonist also directly acts on breast tumour associated fibroblasts to downregulate nuclear factor-κB pathway and metalloproteinases ( P08253 and P14780 ) expression and activity . In conclusion , NRs agonists disrupt the inflammatory cross-talk of the hypoxic breast CSCs niche . 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 . Green tea epigallocatechin-3-gallate attenuates Porphyromonas gingivalis-induced atherosclerosis . The purpose of this study was to determine whether epigallocatechin-3-gallate ( EGCG ) ameliorates Porphyromonas gingivalis-induced atherosclerosis . EGCG is a polyphenol extract from green tea with health benefits and P. gingivalis is shown here to accelerate atheroma formation in a murine model . P02649 knockout mice were administered EGCG or vehicle in drinking water ; they were then fed high-fat diets and injected with P. gingivalis three times a week for 3 weeks . Mice were then killed at 15 weeks . Atherosclerotic plaques in the proximal aorta were determined by Oil Red O staining . Atherosclerosis risk factors in serum , liver or aorta were analysed using cytokine antibody arrays , enzyme-linked immunosorbent assay and real-time PCR . Atherosclerotic lesion areas of the aortic sinus caused by P. gingivalis infection decreased in EGCG-treated groups , wherein EGCG reduced the production of P02741 , monocyte chemoattractant protein-1 , and oxidized low-density lipoprotein ( LDL ) , and slightly lowered LDL/very LDL cholesterol in P. gingivalis-challenged mice serum . Furthermore , the increase in P13500 , P14780 , P05362 , HSP60 , P16070 , P78380 , NOX-4 , P13498 and P35228 gene expression levels in the aorta of P. gingivalis-challenged mice were reduced in EGCG-treated mice . However , P09601 mRNA levels were elevated by EGCG treatment , suggesting that EGCG , as a natural substance , inhibits P. gingivalis-induced atherosclerosis through anti-inflammatory and antioxidative effects . Prion protein stimulates tissue-type plasminogen activator-mediated plasmin generation via a lysine-binding site on kringle 2 . Recombinant human prion-protein ( PrP23-231 ) stimulates plasminogen activation by tissue-type plasminogen activator ( t-PA ) . The stimulatory activity is conserved in the N-terminal fragment ( PrP23-110 ) . It has further been shown by others that P04156 (c) binds to kringle-domains of plasminogen . We compared the stimulatory activity of recombinant PrP23-231 and PrP23-110 on plasminogen activation catalyzed by t-PA , urokinase ( u-PA ) , streptokinase and Desmodus salivary plasminogen activator ( DSPAalpha1 ) . As these plasminogen activators are distinct , with respect to their kringle domains we studied their binding to immobilized PrP23-110 . P00747 activation was measured in a chromogenic assay in vitro and binding studies were carried out using surface plasmon resonance technology . We found that recombinant full-length prion protein , PrP23-231 , and PrP23-110 specifically stimulate t-PA mediated plasminogen activation . Two hundred nanomoles per liter of PrP23-110 stimulated 1.8 nmol L(-1) t-PA 48-fold , 180 nmol L(-1) DB04925 (alpha1) 2.5-fold , 1.8 nmol L(-1) u-PA 1.1-fold , and 1.8 nmol L(-1) streptokinase 1.8-fold . Our data show no specific binding for streptokinase . In contrast all plasminogen activators carrying a kringle domain bound to PrP23-110 . We further studied the effect of lysine on binding to PrP23-110 and on plasminogen activation by DB04925 (alpha1) or t-PA . DB00123 decreased both the binding of t-PA to PrP23-110 and the stimulation of plasmin generation by t-PA . Both binding and plasminogen activation of DB04925 (alpha1) were not influenced by the presence of lysine . All plasminogen activators tested bearing kringle domains bind to PrP23-110 . Binding to PrP23-110 is not sufficient for stimulation of plasmin generation . Thus the lysine-binding site of kringle 2 that is unique to t-PA appears to mediate the specific stimulation of plasminogen activation by the cellular prion protein . Synthesis and biological evaluation of novel oxindole-based RTK inhibitors as anti-cancer agents . Given that receptor tyrosine kinases ( RTKs ) have emerged as key regulators of all aspects of cancer development , including proliferation , invasion , angiogenesis and metastasis , the RTK family represents an important therapeutic target for anti-cancer drug development . Oxindole structure has been used in RTK inhibitors such as DB02058 and intedanib . In this study , two series of new heterocyclic compounds containing oxindole scaffold have been designed and synthesized , and their inhibitory activity against the proliferation of nine cancer cell lines has been evaluated . Among them , compounds 9a and 9b displayed the strongest anti-proliferative activity with the IC50s below 10μM . Flow cytometric analysis showed that the compounds 9a and 9b dose-dependently arrested the cell cycle at G0/ P55008 phase . Although the leading compounds DB02058 and intedanib targets P11362 , the kinase activity test revealed that these compounds only showed slight inhibitory activity on P11362 kinase . Further enzymatic test aided by molecular docking simulation in the DB00171 -binding site demonstrated that 9a and 9b are potent inhibitors of c-Kit kinase . These compounds are worthy of further evaluation as anticancer agents . 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 . DB00644 antagonist in the management of prostate cancer . DB00044 -releasing hormone ( P01148 ) agonist therapy to induce medical castration has become the most common form of hormonal therapy for advanced and metastatic prostate cancer . When treatment is started , P01148 agonists initially stimulate the release of LH , causing a surge in serum testosterone that can precipitate a " flare " phenomenon or worsening of disease , particularly in patients with bone metastatic disease . DB00644 ( DB00644 ) receptor antagonism represents a newer approach to medical castration . DB00106 is a pure P30968 antagonist that is devoid of any P01148 agonist activity . Results from 1 phase II and 3 phase III clinical trials demonstrate that abarelix produces medical castration more quickly and without causing testosterone surge , as compared with P01148 agonists with or without a nonsteroidal antagonist . The safety profile in terms of adverse events is comparable between the 2 types of treatment , but the lack of testosterone surge with abarelix might confer a safety advantage by abolishing the risk of a disease flare . Effects of interleukin-2 ( P60568 ) on human plasma lipid , lipoprotein , and P02741 . Six patients with confirmed malignant disease received four consecutive weekly cycles of human recombinant interleukin-2 ( P60568 ) 4 days/week , continuous iv. infusion , 3 X 10(6) U/m2/day . Plasma cholesterol decreased a mean of 7 % within 24 hours after P60568 infusion and decreased by 33 % within 4 days . Plasma cholesterol was significantly lower than baseline concentration by day 21 ( -21 % ) , and day 25 ( -41 % ) was significantly lower than day 21 . Decreased plasma cholesterol was the result of decreased HDL and LDL cholesterol concentrations . Plasma triglyceride demonstrated a mean increase of 46 % after 4 days of therapy and remained greater than baseline concentrations at all time points analyzed . Apolipoprotein AI and AII decreased concomitantly with HDL-cholesterol concentrations , whereas apolipoprotein B after an initial mean decrease of 17 % during the first cycle was not significantly different from baseline during the fourth cycle . P02649 and P08519 were not significantly affected by P60568 treatment . Plasma P02741 ( CRP ) increased by 79 % within 24 hours of therapy , increased by 254 % on day 4 , then decreased to baseline concentrations by day 21 after 3 days off of P60568 . Day 25 CRP was elevated compared to both baseline and day 21 concentrations . P60568 induced plasma lipoprotein changes may be due in part to the induction of interferon gamma .

MH_train_1190

MH_train_1190

MH_train_1190

interacts_with DB00086?

multiple_choice

Interference of NSAIDs with the thrombocyte inhibitory effect of aspirin : a placebo-controlled , ex vivo , serial placebo-controlled serial crossover study . PURPOSE : Nonsteroidal anti-inflammatory drugs ( NSAIDs ) and acetylsalicylic acid ( ASA ) are often prescribed concurrently in patients with nociceptive pain and cardiovascular comorbidity . NSAIDs and ASA inhibit the same P36551 -enzymes , and thus may interact . ASA 's cardioprotective antiplatelet effect is entirely P23219 dependent . NSAIDs can be either non- P23219 and P35354 selective or P35354 selective . The aim of this study was to examine the interaction between ASA and different selective and nonselective NSAIDs on thrombocyte function . METHODS : Single-blind , prospective , placebo-controlled , ex vivo , serial crossover trial of 3-day cycles separated by washout periods of at least 12 days in 30 healthy volunteers , evaluating interaction on ASA 's antithrombocyte effect by naproxen , ibuprofen , meloxicam , or etoricoxib taken 2 h before ASA . Ex vivo thrombocyte function , closure time ( CT ) in seconds , was measured using the Platelet Function Analyzer 100 ( PFA-100 ) . CT prolongation during a cycle reflects thrombocyte inhibitory effect . ASA nonresponse was defined as CT prolongation < 40 % in the placebo cycle . ASA nonresponders were excluded . Wilcoxon signed-rank was used to evaluate NSAID effect on ASA-induced CT prolongation . RESULTS : Ibuprofen and naproxen inhibit ASA 's antithrombocyte effect below the nonresponse threshold . DB01628 and meloxicam do not cause relevant change in ASA thrombocyte inhibition . Naproxen has an inherent weak thrombocyte inhibitory action below the ASA response threshold . CONCLUSIONS : P23219 affinity determines the interaction between NSAIDs and ASA on thrombocyte adhesion and aggregation . Ibuprofen and naproxen , but not etoricoxib or meloxicam , taken 2 h before ASA , significantly inhibit ASA 's antithrombocyte effect . P00747 /plasmin regulates c-fos and egr-1 expression via the MEK/ P29323 pathway . In this study , we showed that plasminogen ( Plg ) and plasmin ( Pla ) bind to lysine-binding sites on cell surface and trigger a signaling pathway that activates the mitogen-activated protein kinase ( MAPK ) MEK and P27361 /2 , which in turn leads to the expression of the primary response genes c-fos and early growth response gene egr-1 . Our data show that the Plg/Pla-stimulated steady-state mRNA levels of both genes reached a maximum by 30 min and then returned to basal levels by 1h . The gene induction was sensitive to both pharmacological and genetic inhibition of MEK . Leupeptin , a serine protease inhibitor , suppressed Pla but not Plg-induced c-fos and egr-1 expression , emphasizing the role played by the serine protease activity associated with Pla . Pre-incubation with cholera toxin completely blocked the Plg/Pla-induced gene expression , suggesting that another signaling pathway , which recruits G protein-coupled receptors , may also be involved . Furthermore , Plg/Pla also stimulated AP-1 and P18146 DNA-binding activities , which were abrogated by pharmacological inhibition of MEK . Altogether , these results suggest that Plg/Pla stimulates c-fos and egr-1 expression via activation of the MEK/ P29323 pathway . Modification of the reactivity of three amino-acid residues in elongation factor 2 during its binding to ribosomes and translocation . The accessibility of three amino acids of P13639 , located within highly conserved regions near the N- and C-terminal extremities of the molecule ( the E region and the DB02059 region , respectively ) to modifying enzymes has been compared within nucleotide-complexed P13639 and ribosomal complexes that mimic the pre- and posttranslocational ones : the high-affinity complex ( P13639 ) -nonhydrolysable GTP analog GuoPP[CH2]P ribosome and the low-affinity ( P13639 ) -GDP-ribosome complex , P13639 and ribosomes being from rat liver . We studied the reactivity of two highly conserved residues diphthamide-715 and DB00125 -66 , to diphtheria-toxin-dependent ADP-ribosylation and trypsin attack , and of a threonine that probably lies between residues 51 and 60 , to phosphorylation by a Ca2+/calmodulin-dependent protein kinase . DB03223 715 and this threonine residue were unreactive within the high-affinity complex but seemed fully reactive in the low-affinity complex . DB00125 -66 was resistant to trypsin in both complexes . The possible involvement of the E and DB02059 regions of P13639 in the interaction with ribosome in the two complexes is discussed . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . CCK1-receptor stimulation protects against gut mediator-induced lung damage during endotoxemia . BACKGROUND/AIMS : Cholecystokinin 1-receptor ( P32238 ) activation by long chain fatty acid ( LCFA ) absorption stimulates vago-vagal reflex pathways in the brain stem . The present study determines whether this reflex also activates the cholinergic anti-inflammatory pathway , a pathway known to modulate cytokine release during endotoxemia . METHODS : Mesenteric lymph was obtained from wild type ( WT ) and P32238 knockout ( P32238 (-/-) ) mice intraperitoneally challenged with Lipopolysaccharid ( LPS ) ( endotoxemic lymph , EL ) and intestinally infused with vehicle or LCFA-enriched solution . The lymph was analyzed for TNFα , P05231 and P22301 concentration and administered to healthy recipient mice via jugular infusion . Alveolar wall thickness , myeloperoxidase ( P05164 ) and TUNEL positive cells were determined in lung tissue of recipient mice . RESULTS : LCFA infusion in WT mice reduced TNFα concentration in EL by 49 % compared to vehicle infusion , but had no effect in P32238 (-/-) mice . EL significantly increased the alveolar wall thickness , the number of P05164 -positive and TUNEL-positive cells compared to control lymph administration . LCFA infusion in WT , but not in CCK1R(-/-) mice , significantly reduced these pathological effects of EL . CONCLUSION : During endotoxemia enteral LCFA absorption reduces TNFα release into mesenteric lymph and attenuates histomorphologic parameters of lung dysfunction . Failure to elicit this effect in CCK1R(-/-) mice demonstrates that anti-inflammatory properties of LCFAs are mediated through CCK1-Rs . Benzo[b]thiophene-based histone deacetylase inhibitors . Benzo[b]thienyl hydroxamic acids , a novel class of histone deacetylase ( HDAC ) inhibitors , were identified via a targeted screen of small molecule hydroxamic acids . Various substitutions were explored in the P01031 - and P13671 -positions of the benzo[b]thiophene core to characterize SAR and develop optimal inhibitors . It was determined that substitution at the P13671 -position of the benzo[b]thiophene core with a three-atom spacer yielded optimal Q13547 inhibition and anti-proliferative activity in murine erythroleukemia ( SC-9 ) cells . P15121 regulates TGF-beta1-induced production of fibronectin and type IV collagen in cultured rat mesangial cells . AIM : To study the effects of aldose reductase ( AR ) on production of fibronectin and type IV collagen in rat mesangial cells ( MsC ) . METHODS : The vector , pcDNA3-AR , was constructed based on pET-15b-AR . Lipofect AMINE was used for stable transfection and G418 was used for selecting positive clones . DB02712 and zopolrestat were added for suppressing the activity of AR , respectively . The production of fibronectin and type IV collagen and the activation of Smads and MAPK signal transduction pathway were analysed by western blot and AP-1 activity was analysed by electrophoretic mobility shift assays ( EMSA ) . RESULTS : The normal MsC showed increased expression of fibronectin and type IV collagen with stimulation of TGF-beta1 . Compared with the normal MsC , the MsC pre-incubated with Q9Y4X5 showed reduced expression ( P < 0.05 ) and the AR-transfected MsC showed increased expression ( P < 0.05 ) . The normal MsC showed activation of P29323 , JNK and p38 with stimulation of TGF-beta1 , while the activation of JNK and p38 was inhibited in the MsC pre-incubated with Q9Y4X5 and only the activation of JNK was enhanced in the AR-transfected MsC . The normal MsC showed enhanced AP-1 activity with the stimulation of TGF-beta1 , and similarly the activity was inhibited in the MsC pre-incubated with Q9Y4X5 and was more enhanced in the AR transfected MsC . CONCLUSION : AR can regulate the expression of fibronectin and type IV collagen with the stimulation of TGF-beta1 in MsC , which may have relations with the activation of JNK-MAPK and p38-MAPK signalling pathways and AP-1 . Selective expression of the large neutral amino acid transporter at the blood-brain barrier . Amino acid supply in brain is regulated by the activity of the large neutral amino acid transporter ( O43561 ) at the brain capillary endothelial cell , which forms the blood-brain barrier ( BBB ) in vivo . Bovine BBB poly(A)(+) RNA was isolated from 2.0 kg of fresh bovine brain and size fractionated on a sucrose density gradient , and a size-fractionated bovine BBB cDNA library in the pSPORT vector was prepared . The full-length cDNA encoding the bovine BBB O43561 was isolated from this library , and the predicted amino acid sequence was 89-92 % identical to the Q01650 isoform . The bovine BBB Q01650 mRNA produced a 10-fold enhancement in tryptophan transport into frog oocytes coinjected with bovine BBB Q01650 mRNA and the mRNA for P08195 , which encodes the heavy chain of the heterodimer . DB00150 transport into the mRNA-injected oocytes was sodium independent and was specifically inhibited by other large neutral amino acids , and the K(m) of tryptophan transport was 31.5 +/- 5.5 microM . Northern blotting with the bovine BBB Q01650 cDNA showed that the Q01650 mRNA is 100-fold higher in isolated bovine brain capillaries compared with P13671 rat glioma cells or rat brain , and the Q01650 mRNA was not detected in rat liver , heart , lung , or kidney . These studies show that the Q01650 transcript is selectively expressed at the BBB compared with other tissues , and the abundance of the Q01650 mRNA at the BBB is manyfold higher than that of transcripts such as the P08195 antigen , actin , or the Glut1 glucose transporter . [ 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 . Phosphorylation of elongation factor 2 in normal and malignant rat glial cells . Certain calmodulin ( P62158 ) -dependent protein kinases phosphorylate substrates have been implicated in regulating cellular proliferation . In this study , P62158 -dependent phosphorylation has been examined in normal and tumor tissue from rat brain to determine whether differences exist . Using in vitro phosphorylation reactions , we compared endogenous substrates for Ca2+/ P62158 -dependent protein kinases in rat brain white matter ( RBWM ) , a tissue rich in normal glia , to those of P13671 rat glioma cells . A major phosphoprotein having a M(r) of 100,000 was observed in proliferating P13671 cells that was not present in RBWM or in nonproliferating cells . Phosphorylation was stimulated by Ca2+ and P62158 and inhibited by trifluoperazine . An antibody to elongation factor 2 ( P13639 ) immunoprecipitated the M(r) 100,000 protein from P13671 cells . P13639 was present in RBWM but was not phosphorylated . Homogenates of RBWM did not phosphorylate exogenous P13639 , which suggested the absence of P62158 kinase III activity in normal glial tissue . Furthermore , the addition of purified , exogenous P62158 kinase III to homogenates of RBWM resulted in P13639 phosphorylation . These data demonstrate that a basal level of P13639 phosphorylation exists in proliferating glioma cells that is markedly diminished or absent in normal glial tissue and is due to the activity of P62158 kinase III . 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 . Pharmacodynamics and pharmacokinetics of omapatrilat in heart failure . The purpose of this study was to determine the pharmacodynamics and pharmacokinetics of omapatrilat , administered orally ( 25 mg ) or intravenously ( 10 mg ) in 19 New York Heart Association class II and class III congestive heart failure ( CHF ) patients versus 17 healthy controls matched for age , race , gender , and weight . The plasma concentrations of atrial natriuretic peptide ( P01160 ) increased by approximately 20 % and 30 % in CHF and control subjects , respectively , at 4 hours after intravenous or oral omapatrilat administration . Similar elevation in the cyclic guanosine monophosphate concentration ( 25 % to 35 % ) and P01160 urinary excretion ( 21 ng/24 h to 22 ng/24 h ) was seen in all treatment groups after omapatrilat administration . P12821 activity was > 90 % inhibited at 4 hours after dosing and remained approximately 60 % to 70 % inhibited at 24 hours after dosing . The levels of endothelin-1 and endothelin-2 remained unchanged after oral or intravenous administration of omapatrilat . The maximal reduction in seated blood pressure compared with baseline was similarfor CHF and control subjects . Clinical pharmacokinetic parameters were similar in both groups after intravenous dosing , but maximum concentration and area under the concentration-time curve were elevated in CHF patients compared with controls after oral dosing . DB00886 was well tolerated ; differences in systemic exposure and metabolism between CHF patients and controls did not appear to be clinically significant . 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 . Q96NZ9 -sensitive P15309 -conjugated nanoparticles for multi-targeting therapy of brain glioma . Now it is well evidenced that tumor growth is a comprehensive result of multiple pathways , and glioma parenchyma cells and stroma cells are closely associated and mutually compensatory . Therefore , drug delivery strategies targeting both of them simultaneously might obtain more promising therapeutic benefits . In the present study , we developed a multi-targeting drug delivery system modified with uPA-activated cell-penetrating peptide ( P15309 ) for the treatment of brain glioma ( P01160 ) . In vitro experiments demonstrated nanoparticles ( NP ) decorated with cell-penetrating peptide ( CPP ) or P15309 could significantly improve nanoparticles uptake by P13671 glioma cells and nanoparticles penetration into glioma spheroids as compared with traditional NP and thus enhanced the therapeutic effects of its payload when paclitaxel ( PTX ) was loaded . In vivo imaging experiment revealed that P01160 accumulated more specifically in brain glioma site than NP decorated with or without CPP . Brain slides further showed that P15309 contributed to more nanoparticles accumulation in glioma site , and P01160 could co-localize not only with glioma parenchyma cells , but also with stroma cells including neo-vascular cells and tumor associated macrophages . The pharmacodynamics results demonstrated P15309 could significantly improve the therapeutic benefits of nanoparticles by significantly prolonging the survival time of glioma bearing mice . In conclusion , the results suggested that nanoparticles modified with uPA-sensitive P15309 could reach multiple types of cells in glioma tissues and provide a novel strategy for glioma targeted therapy . 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 . DB05651 , a novel isotype-selective histone deacetylase inhibitor , has broad spectrum antitumor activity in vitro and in vivo . Nonselective inhibitors of human histone deacetylases ( HDAC ) are known to have antitumor activity in mice in vivo , and several of them are under clinical investigation . The first of these , DB02546 ( DB02546 ) , has been approved for treatment of cutaneous T-cell lymphoma . Questions remain concerning which HDAC isotype(s) are the best to target for anticancer activity and whether increased efficacy and safety will result with an isotype-selective HDAC inhibitor . We have developed an isotype-selective HDAC inhibitor , DB05651 , which potently targets human Q13547 but also has inhibitory activity against Q92769 , O15379 , and Q96DB2 in vitro . In intact cells , DB05651 inhibited only a fraction of the total HDAC activity and showed long-lasting inhibitory activity even upon drug removal . DB05651 induced hyperacetylation of histones , selectively induced apoptosis , and caused cell cycle blockade in various human cancer cell lines in a dose-dependent manner . DB05651 exhibited potent and selective antiproliferative activities against a broad spectrum of human cancer cell lines in vitro , and HDAC inhibitory activity was required for these effects . In vivo , DB05651 significantly inhibited growth of human tumor xenografts in nude mice in a dose-dependent manner and the antitumor activity correlated with induction of histone acetylation in tumors . Our findings suggest that the isotype-selective HDAC inhibition by DB05651 is sufficient for antitumor activity in vivo and that further clinical investigation is warranted . Enhancement of L-cystine transport activity and its relation to Q9UPY5 gene induction at the blood-brain barrier by diethyl maleate treatment . The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier ( BBB ) . The uptake of [(14)C]L-cystine and [(3)H]L-glutamic acid ( L- DB00142 ) was determined using a mouse brain endothelial cell line ( MBEC4 ) as an in vitro BBB model . The mRNA levels of L-cystine/L- DB00142 exchanger , system x(c)(-) , which consists of Q9UPY5 and P08195 , were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis . The [(14)C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na(+)-independent saturable process . The corresponding Michaelis-Menten constant ( K(m) ) was 63.7 microM . In the presence of L- DB00142 , there was competitive inhibition with an inhibition constant ( K(i) ) of 83.5 microM . [(3)H]L- DB00142 uptake in the absence of Na(+) was saturable with a K(m) of 48.1 microM , and it exhibited competitive inhibition with a K(i) of 24.9 microM in the presence of L-cystine . The mutual inhibition between L-cystine and L- DB00142 and the type of inhibition suggest that system x(c)(-) operates in MBEC4 cells . The Q9UPY5 and P08195 mRNAs were expressed in MBEC4 cells and , following diethyl maleate ( DEM ) treatment , the Q9UPY5 mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration ( up to 500 microM ) . Concomitantly , the glutathione concentration in MBEC4 cells was increased . In conclusion , system x(c)(-)-mediated L-cystine uptake takes place in MBEC4 cells . DB00138 transport via system x(c)(-) at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the Q9UPY5 gene . DB01093 -treated HL60 cells : a model of neutrophil-like cells mainly expressing Q07343 subtype . The human promyelocytic HL60 cells acquired a neutrophilic phenotype after a 7- to 10-day DB01093 treatment . Fc gammaRII was up-regulated . Fc gammaRI was also up-regulated by an additional P01579 treatment . These cells are able to produce O2*- by NADPH oxidase activation in the presence of immune complexes or phorbol-12-myristate-13-acetate ( PMA ) . A change of their DB05876 subtype profile was also observed : Q07343 was the predominant isoenzyme , Q08499 was down-regulated and P27815 was no longer detectable . Additionally , the more NADPH oxidase was activated by PMA , the less P27815 was expressed , suggesting that NADPH oxidase activity could be used as a surrogate marker of P27815 down-regulation . DB01954 and DB03849 ( cilomilast ) , two selective DB05876 inhibitors , dose-dependently inhibited receptor-coupled activation of superoxide . These results suggest that Q07343 is the main subtype involved in regulating superoxide induced by Fc gammaRs activation . Furthermore , these cells , expressing almost exclusively Q07343 subtype , could be useful to identify selective Q07343 inhibitors . PDE4B5 , a novel , super-short , brain-specific DB02527 phosphodiesterase-4 variant whose isoform-specifying N-terminal region is identical to that of DB02527 phosphodiesterase-4D6 ( PDE4D6 ) . The DB02527 -specific phosphodiesterase-4 ( DB05876 ) gene family is the target of several potential selective therapeutic inhibitors . The four DB05876 genes generate several distinct protein-coding isoforms through the use of alternative promoters and 5'-coding exons . Using mouse transcripts , we identified a novel , super-short isoform of human Q07343 encoding a novel 5' terminus , which we label PDE4B5 . The protein-coding region of the novel 5' exon is conserved across vertebrates , chicken , zebrafish , and fugu . Reverse-transcription-polymerase chain reaction ( PCR ) and quantitative ( PCR ) measurements show that this isoform is brain-specific . The novel protein is 58 +/- 2 kDa ; it has DB02527 hydrolyzing enzymatic activity and is inhibited by DB05876 -selective inhibitors rolipram and cilomilast ( DB03849 ) . Confocal and subcellular fractionation analyses show that it is distributed predominantly and unevenly within the cytosol . The 16 novel N-terminal residues of PDE4B5 are identical to the 16 N-terminal residues of the super-short isoform of Q08499 ( PDE4D6 ) , which is also brain-specific . PDE4B5 is able to bind the scaffold protein Q9NRI5 , whose gene has been linked to schizophrenia . Microarray expression profiling of the DB05876 gene family shows that specific DB05876 genes are enriched in muscle and blood fractions ; however , only by monitoring the individual isoforms is the brain specificity of the super-short Q08499 and Q07343 isoforms revealed . Understanding the distinct tissue specificity of DB05876 isoforms will be important for understanding phosphodiesterase biology and opportunities for therapeutic intervention . Differential effects of P47900 and Q9H244 nucleotide receptors on P27361 / P28482 and phosphatidylinositol 3-kinase signalling and cell proliferation in serum-deprived and nonstarved glioma P13671 cells . We have previously shown that , in glioma P13671 cells , two nucleotide ADP-sensitive receptors coexist : P47900 , coupled to P98160 and responsible for Ca2+ release , and Q9H244 , negatively coupled to adenylate cyclase . In the present study , we examined the effects of the stimulation of these two receptors on P27361 /2 and P19957 -K activation , and cell proliferation in either serum-deprived or nonstarved P13671 cells . In response to ADP and its analogues , in serum-starved cells , both Q8TCB0 P27361 and Q8NFH3 P28482 were activated in a time-dependent manner , as monitored by Western blot analysis using an antiphospho- Q8NFH3 / Q8TCB0 MAPK antibody . The phosphorylation was reduced both by removal of the extracellular Ca2+ and partially or almost completely by MRS2179 or AR-C69931MX , specific antagonists of the P47900 and Q9H244 receptors , respectively . The inhibitory effect of antagonists was additive . These data indicate the involvement of both receptors , P47900 and Q9H244 , in the P27361 /2 activation , but the Q9H244 receptor contribution predominates . P27361 /2 activity was positively correlated with cell proliferation of cultured glioma P13671 cells . In nonstarved cells , ADP markedly decreased the P19957 -K activity . In contrast , in serum-starved cells , ADP evoked an increase in the P19957 -K activity . Blocking of the P47900 receptor by MRS2179 additionally increased this ADP response . These results suggest that the P47900 receptor has an inhibitory and the Q9H244 receptor a stimulatory effect on P19957 -K signalling pathway . RT-PCR analysis revealed different mRNA expression of both receptors in starved and nonstarved cells . In nonstarved cells , the P47900 receptor mRNA predominates , whereas in serum-deprived cells the expression of Q9H244 mRNA becomes more pronounced . British Journal of Pharmacology ( 2004 ) 141 , 497-507. doi:10.1038/sj.bjp.0705639 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 . Possible role of cholecystokinin-A receptors in regulation of thyrotropin ( DB00024 ) secretion in male rats . We studied the importance of cholecystokinin ( CCK ) system in the regulation of thyrotropin ( DB00024 ) and prolactin ( PRL ) secretion in male rats . To this end , we tested the effects of both unselective CCK agonists CCK-8 and caerulein , and CCK-B selective agonists Q13308 and pentagastrin as well as the selective CCK antagonists ( devazepide and L-365,260 ) at wide dose-ranges on the cold-stimulated and TRH-induced DB00024 and PRL secretion . DB00403 , given s.c . 15 min before sacrifice , decreased DB00024 levels at 5 micrograms/kg . In time course-studies , the maximum inhibition was seen at 15 min but the effect lasted at least 30 , but less than 60 min . Also CCK-8 decreased DB00024 levels at the doses of 20 and 50 micrograms/kg at 15 min . Devazepide and L-365,260 did not affect DB00024 or PRL levels at any dose . The effect of caerulein ( 5 micrograms/kg ) was antagonized by devazepide , a CCK-A antagonist , at 100 micrograms/kg , but not by a CCK-B antagonist L-365,260 tested at a wide dose range . PRL levels were not affected by any treatment . DB00403 ( 5 micrograms/kg ) , given at the same time as TRH ( 500 ng/kg ) , inhibited the TRH-induced DB00024 levels at 15 min , but not at 30 or 60 min . CCK-8 ( 50 micrograms/kg ) , Q13308 ( 100 micrograms/kg ) and pentagastrin ( 500 micrograms/kg ) did not affect the TRH-induced DB00024 secretion . The results probably indicate that P32238 stimulation inhibits DB00024 secretion at the level of the anterior pituitary gland . PRL levels in male rats are not affected by CCK system .

MH_train_1191

MH_train_1191

MH_train_1191

interacts_with DB09068?

multiple_choice

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 . 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 . Rectal antinociceptive properties of alverine citrate are linked to antagonism at the P08908 receptor subtype . Serotonin ( 5-HT ) is considered as a major mediator causing hyperalgesia and is involved in inflammatory reactions and irritable bowel syndrome . DB01616 citrate may possess visceral antinociceptive properties in a rat model of rectal distension-induced abdominal contractions . This study was designed to evaluate the pharmacological properties of alverine citrate in a rat model of rectal hyperalgesia induced by 5-HTP ( 5-HT precursor ) and by a selective P08908 agonist ( 8-OH-DPAT ) and to compare this activity with a reference P08908 antagonist ( WAY 100635 ) . At 4 h after their administration , 5-HTP and 8-OH-DPAT increased the number of abdominal contractions in response to rectal distension at the lowest volume of distension ( 0.4 mL ) . When injected intraperitoneally before 8-OH-DPAT and 5-HTP , WAY 100635 ( 1 mg kg(-1) ) blocked their nociceptive effect , but also reduced the response to the highest volume of distension ( 1.6 mL ) . Similarly , when injected intraperitoneally , alverine citrate ( 20 mg kg(-1) ) suppressed the effect of 5-HTP , but not that of 8-OH-DPAT . However , when injected intracerebroventricularly ( 75 microg/rat ) alverine citrate reduced 8-OH-DPAT-induced enhancement of rectal distension-induced abdominal contractions . In-vitro binding studies revealed that alverine citrate had a high affinity for P08908 receptors and a weak affinity for 5- Q9H205 and Q13639 subtypes . These results suggest that 5-HTP-induced rectal hypersensitivity involves 5-TH1A receptors and that alverine citrate acts as a selective antagonist at the P08908 receptor subtype to block both 5-HTP and 8-OH-DPAT-induced rectal hypersensitivity . Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . Evidence that 5-HT2c receptor antagonists are anxiolytic in the rat Geller-Seifter model of anxiety . Four non-selective P28335 /5- Q13049 receptor antagonists , mianserin ( 2-8 mg/kg ) , 1-naphthyl piperazine ( 1-NP ) ( 0.5-1 mg/kg ) , ICI 169,369 ( 20 mg/kg ) and LY 53857 ( 5 mg/kg ) , increased punished responding for a food reward in the rat Geller-Seifter test 30 min after subcutaneous ( SC ) administration . This property was shared by the benzodiazepine anxiolytic chlordiazepoxide ( 5 mg/kg SC ) . However , the selective 5- Q13049 receptor antagonists ketanserin ( 0. P35326 mg/kg SC ) and altanserin ( 0.5 , 1 mg/kg SC ) had little effect . The P08908 , P28222 and beta-adrenergic receptor antagonists pindolol and cyanopindolol ( 6 mg/kg SC ) did not affect punished responding either , nor did the P28221 receptor partial agonist and alpha 2 adrenergic receptor antagonist yohimbine ( 2.5 mg/kg SC ) or the histamine H1 receptor antagonist mepyramine ( 1 mg/kg SC ) . Unpunished responding was also modestly increased after some doses of the P28335 /5- Q13049 receptor antagonists . However , this effect was inconsistent and was also seen after chlordiazepoxide . Furthermore , it was not associated with the increase in punished responding observed in rats orally treated with mianserin ( 10 , 20 mg/kg ) , 1-NP ( 10 , 20 mg/kg ) or ICI 169,369 ( 50 mg/kg ) . The action of the P28335 /5- Q13049 receptor antagonists tested is therefore consistent with anxiolysis . The results also strongly suggest that this effect is mediated by blockade of the P28335 receptor , although the possibility of P41595 receptor mediation is discussed . 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 . Mutational analysis of the mitochondrial P47985 of Saccharomyces cerevisiae . II . Biochemical characterization of temperature-sensitive Q13546 - mutations . Although the function of the P47985 is generally understood , little is known of how the structure of this protein supports its mechanistic role in electron transfer in the cytochrome bc1 complex . To better understand the structural basis of iron-sulfur protein function , we have undertaken a mutational analysis of the gene encoding this protein and initially isolated five temperature-sensitive iron-sulfur protein mutants ( Beckmann , J. D. , Ljungdahl , P. O. , and Trumpower , B. L. ( 1989 ) J. Biol. Chem. 264 , 3713-3722 ) . Each of the five ts-rip1- mutants exhibited pleiotropic effects . Although the mutant iron-sulfur proteins manifest several in vitro phenotypes in common , each exhibited unique characteristics . All of the ts-rip1- mutations resulted in membranes with decreased ubiquinol-cytochrome c oxidoreductase activities and decreased thermostability compared to membranes containing wild type iron-sulfur protein . All of the mutations conferred slight but significant resistance to the respiratory inhibitor myxothiazol , and one mutant was hypersensitive to inhibition by DB04799 , a structural analog of ubiquinone . In addition , one of the mutations completely blocks post-translational processing of the iron-sulfur protein , leading to accumulation of pre-iron-sulfur protein in mitochondrial membranes at nonpermissive temperatures . Finally , a mutation 12-amino acid residues away from the carboxyl terminus ( 203S ) results in an extremely unstable protein . This region of the protein may be essential in blocking degradation of pre-iron-sulfur protein by cytoplasmic proteases as the protein is imported into the mitochondria , or may be a " degradation signal , " which tags the iron-sulfur protein for turnover . DB05332 administration shows reduced megakaryocyte response-capacity and increased myelofibrosis in a mouse model of P35579 -RD . Macrothrombocytopenia in P35579 -related disease ( P35579 -RD ) results from defects in nonmuscular myosin-IIA function . P40238 agonists ( eltrombopag ; romiplostim ) seem to improve hemostasis , but little is known about their biologic effects in P35579 -RD . We administered romiplostim to Myh9(-/-) mice ( 100 μg/kg , every 3 days , during 1 month ) . MKs increased to similar numbers in Myh9(-/-) and wild-type ( WT ) mice ( with an increase in immature MKs ) , but Myh9(-/-) platelet count response was much less ( 2.5-fold vs 8-fold increase ) . A strong increase in MK nuclei emboli in the lung , in WT and Myh9(-/-) mice , indicates increased transmigration of MKs from the BM . Prolonged ( but not acute ) treatment with romiplostim decreased expression of GPIb-IX-V complex and Q9HCN6 , but not of GPIIbIIIa , and bleeding time increased in WT mice . Microcirculation was not altered by the increased number of large platelets in any of the assessed organs , but in Myh9(-/-) mice a much stronger increase in BM reticulin fibers was present after 4 weeks of romiplostim treatment vs WT mice . These data further encourage short-term use of thrombopoietic agents in patients with P35579 -RDs ; however , myelofibrosis has to be considered as a potential severe adverse effect during longer treatment . Reduction of GPIbIX/ Q9HCN6 expression by romiplostim requires further studies . 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 . Proteolytic cleavage of platelet endothelial cell adhesion molecule-1 ( P16284 /CD31 ) is regulated by a calmodulin-binding motif . Homophilic engagement of platelet endothelial cell adhesion molecule-1 ( P16284 /CD31 ) induces ' outside-in ' signal transduction that results in phosphorylation events and recruitment and activation of signalling molecules . The formation of signalling scaffolds with P16284 are important signalling events that modulate platelet secretion , aggregation and platelet thrombus formation . In this study , we describe a novel interaction between P16284 and cytosolic calmodulin ( P62158 ) in platelets . Reciprocal co-immunoprecipitation studies revealed that cytosolic P62158 is constitutively associated with P16284 in resting , thrombin activated and aggregated human platelets . Our studies demonstrate that P62158 directly interacts with a P16284 peptide ( 594-604 ) C595A containing the sequences (594)KAFYLRKAKAK(604) . This P62158 : P16284 interaction has a threefold higher affinity than P62158 : Q9HCN6 interaction . It is potentiated by the addition of calcium ions , and dissociated by the P62158 inhibitor , trifluoperazine . Treatment of platelets with P62158 inhibitors triggers cleavage of P16284 in a time- and dose-dependent manner . Furthermore , this membrane proximal portion of P16284 is conserved across mammalian species and the helical representation of basic/hydrophobic residues reveals a charge distribution analogous to other P62158 -binding motifs in other proteins . Taken together , these results suggest that this highly charged cluster of amino acids in the P16284 cytoplasmic domain directly interacts with P62158 and this novel interaction appears to regulate cleavage of P16284 . Adjacent cysteines are capable of ligating the same tetranuclear iron-sulfur cluster . The mechanism of the energy-converting DB00157 ( beta-nicotinamide adenine dinucleotide , reduced form ) :ubiquinone oxidoreductase , which is also called respiratory complex I , is largely unknown due to lack of a high-resolution structure and the most complicated construction of the enzyme . Electron transport is carried out by one flavin mononucleotide ( Q68DA7 ) and up to 9 Fe/S clusters . The Fe/S cluster N2 , which is believed to be directly involved in redox-coupled proton-translocation , is located on subunit NuoB ( the homologue of the mitochondrial O75251 ) . This subunit contains a conserved binding motif for a [ 4Fe/4S ] cluster with two adjacent cysteines . It was questioned whether these adjacent cysteines could be ligands of the same cluster due to a possible steric hinderance . However , mutagenesis of either of these cysteines led to a loss of cluster N2 . We used the known structure of the homologous small subunit of hydrogenases containing a regular cysteine motif to generate an in silico mutant with two consecutive cysteines . Molecular dynamics simulation showed that the conformation of these cysteines does not meet the topological requirements for coordination of a [ 4Fe/4S ] cluster when the protein backbone conformation is kept constant . In comparison , the simulation of a dipeptide amide using a " template forcing " approach resulted in a conformation compatible to an optimal coordination of the two cluster positions in question . Thus , a slight main-chain conformational change would allow two adjacent cysteines to coordinate a [ 4Fe/4S ] cluster . Innovative approaches for the development of antidepressant drugs : current and future strategies . Depression is a highly debilitating disorder that has been estimated to affect up to 21 % of the world population . Despite the advances in the treatment of depression with selective serotonin reuptake inhibitors ( SSRIs ) and serotonin and norepinephrine reuptake inhibitors ( SNRIs ) , there continue to be many unmet clinical needs with respect to both efficacy and side effects . These needs range from efficacy in treatment resistant patients , to improved onset , to reductions in side effects such as emesis or sexual dysfunction . To address these needs , there are numerous combination therapies and novel targets that have been identified that may demonstrate improvements in one or more areas . There is tremendous diversity in the types of targets and approaches being taken . At one end of a spectrum is combination therapies that maintain the benefits associated with SSRIs but attempt to either improve efficacy or reduce side effects by adding additional mechanisms ( P08908 , P28222 , P28221 , P28335 , alpha-2A ) . At the other end of the spectrum are more novel targets , such as neurotrophins ( P23560 , IGF ) , based on recent findings that antidepressants induce neurogenesis . In between , there are many approaches that range from directly targeting serotonin receptors ( P28335 , P50406 ) to targeting the multiplicity of potential mechanisms associated with excitatory ( glutamate , DB01221 , Q14416 , P41594 ) or inhibitory amino acid systems ( GABA ) or peptidergic systems ( neurokinin 1 , DB05394 1 , melanin-concentrating hormone 1 , V1b ) . The present review addresses the most exciting approaches and reviews the localization , neurochemical and behavioral data that provide the supporting rationale for each of these targets or target combinations . 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 . DB00659 : recent findings and future research directions . This article explores the mechanisms of action and the potential responder profile of acamprosate , a compound efficacious in relapse prevention of alcoholism . New evidence at the molecular and cellular level suggests that acamprosate attenuates hyper-glutamatergic states that occur during early abstinence and involves iono ( DB01221 ) - and metabotrotropic ( P41594 ) glutamate receptors along with augmented intracellular calcium release and electrophysiological changes . Thus mutant mice with enhanced glutamate levels exhibit higher alcohol consumption than wild type mice and respond better to acamprosate , demonstrating that acamprosate acts mainly on a hyper-glutamatergic system . This mode of action further suggests that acamprosate exhibits neuroprotective properties . In rats , cue-induced reinstatement behavior is significantly reduced by acamprosate treatment whereas cue-induced craving responses in alcohol-dependent patients seem not to be affected by this treatment . An ongoing study ( " Project Predict " ) defines specific responder profiles for an individualized use of acamprosate and naltrexone . Neurophysiological as well as psychometric data are used to define 2 groups of patients : " reward cravers " and " relief cravers " . While naltrexone should work better in the first group , acamprosate is hypothesized to be efficacious in the latter where withdrawal associated and/or cue induced hyper-glutamatergic states are thought to trigger relapse . Further research should target the definition of subgroups applying endophenotypic approaches , e.g. by detecting a hyperglutamatergic syndrome using MR spectroscopy . Genetic polymorphism and activities of human lung alcohol and aldehyde dehydrogenases : implications for ethanol metabolism and cytotoxicity . DB00898 dehydrogenase ( DB00067 ) and aldehyde dehydrogenase ( ALDH ) exhibit genetic polymorphism and tissue specificity . DB00067 and ALDH isozyme phenotypes from 39 surgical Chinese lung specimens were identified by agarose isoelectric focusing . The identity of the lung beta-ADHs was further demonstrated by their characteristic pH-activity profiles for ethanol oxidation , Km values for NAD and ethanol , and inhibition by DB01213 or 1,10-phenanthroline . The beta 2 allele , coding for beta 2 polypeptide , was found to be predominant in the lung specimens studied . The DB00067 activities in the lungs with the homozygous phenotype P00325 2-2 ( exhibiting beta 2 beta 2 ) and P00325 1-1 ( exhibiting beta 1 beta 1 ) and the heterozygous phenotype P00325 P35326 ( exhibiting beta 2 beta 2 , beta 2 beta 1 , and beta 1 beta 1 ) were determined to be 999 +/- 77 , 48 +/- 17 , and 494 +/- 61 nmol/min/g tissue , respectively . Fifty-one percent of the specimens studied lacked the P05091 activity band on the isoelectric focusing gels . The activities in the lung tissues with the P05091 -active phenotype and the inactive phenotype were determined to be 30 +/- 3 and 17 +/- 1 nmol/min/g tissue , respectively . These findings indicate that human pulmonary ethanol-metabolizing activities differ significantly with respect to genetic polymorphism at both the P00325 and the P05091 loci . The results suggest that individuals with high Vmax beta 2- DB00067 and deficient in low-Km mitochondrial P05091 , accounting for approximately 45 % of the Chinese population , may end up with acetaldehyde accumulation during alcohol consumption , rendering them vulnerable to tissue injury caused by this highly reactive and toxic metabolite . Differential regulation of the serotonin 1 A transcriptional modulators five prime repressor element under dual repression-1 and nuclear-deformed epidermal autoregulatory factor by chronic stress . Chronic stress is known to affect brain areas involved in learning and emotional responses . These changes , thought to be related to the development of cognitive deficits are evident in major depressive disorder and other stress-related pathophysiologies . The serotonin-related transcription factors ( Q6P1N0 / Q6P1N0 ; five prime repressor element under dual repression/coiled-coil P06681 domain 1a , and O75398 /Deaf-1 ; nuclear-deformed epidermal autoregulatory factor ) are two important regulators of the P08908 receptor . Using Western blotting and quantitative real-time polymerase chain reaction ( qPCR ) we examined the expression of mRNA and proteins for Q6P1N0 , O75398 , and the P08908 receptor in the prefrontal cortex ( P27918 ) of male rats exposed to chronic restraint stress ( CRS ; 6 h/day for 21 days ) . After 21 days of CRS , significant reductions in both Q6P1N0 mRNA and protein were observed in the P27918 ( 36.8 % and 32 % , respectively ; P < 0.001 ) , while the levels of both O75398 protein and mRNA did not change significantly . Consistent with reduced Q6P1N0 protein , P08908 receptor mRNA levels were equally upregulated in the P27918 , while protein levels actually declined , suggesting post-transcriptional receptor downregulation . The data suggest that CRS produces distinct alterations in the serotonin system specifically altering Q6P1N0 and the P08908 receptor in the P27918 of the male rat while having no effect on O75398 . These results point to the importance of understanding the mechanism for the differential regulation of Q6P1N0 and O75398 in the P27918 as a basis for understanding the related effects of chronic stress on the serotonin system ( serotonin-related transcription factors ) and stress-related disorders like depression . Reversible defects in O-linked glycosylation and P01130 expression in a UDP-Gal/ Q14376 deficient mutant . We previously isolated an unusual hamster cell mutant ( ldlD ) that does not express P01130 activity unless it is cocultivated with other cells or grown in high concentrations of serum . We now show that ldlD cells are deficient in the enzyme DB03501 and UDP-N-acetylgalactosamine ( GalNAc ) 4-epimerase . When ldlD cells are grown in glucose-based media , they can not synthesize enough DB03501 and UDP-GalNAc to allow normal synthesis of glycolipids and glycoproteins . The 4-epimerase deficiency accounts for all glycosylation defects previously observed in ldlD cells , including production of abnormal LDL receptors . All abnormal phenotypes of ldlD cells can be fully corrected by exogenous galactose and GalNAc . The separate effects of these sugars on P01130 activity suggest that O-linked carbohydrate chains are crucial for receptor stability . ldlD cells may be useful for structural and functional studies of many proteins , proteoglycans , and glycolipids containing galactose or GalNAc . Low-intensity laser irradiation at 660 nm stimulates transcription of genes involved in the electron transport chain . BACKGROUND DATA : Low-intensity laser irradiation ( LILI ) has been shown to stimulate cellular functions leading to increased adenosine triphosphate ( DB00171 ) synthesis . This study was undertaken to evaluate the effect of LILI on genes involved in the mitochondrial electron transport chain ( ETC , complexes I-IV ) and oxidative phosphorylation ( DB00171 synthase ) . METHODS : Four human skin fibroblast cell models were used in this study : normal non-irradiated cells were used as controls while wounded , diabetic wounded , and ischemic cells were irradiated . Cells were irradiated with a 660 nm diode laser with a fluence of 5 J/cm(2) and gene expression determined by quantitative real-time reverse transcription ( RT ) polymerase chain reaction ( PCR ) . RESULTS : LILI upregulated cytochrome c oxidase subunit VIb polypeptide 2 ( Q6YFQ2 ) , cytochrome c oxidase subunit VIc ( P09669 ) , and pyrophosphatase ( inorganic ) 1 ( Q15181 ) in diabetic wounded cells ; P09669 , DB00171 synthase , H+transporting , mitochondrial Fo complex , subunit B1 ( P24539 ) , nicotinamide adenine dinucleotide ( DB00157 ) dehydrogenase ( ubiquinone ) 1 alpha subcomplex , 11 ( Q86Y39 ) , and DB00157 dehydrogenase ( ubiquinone ) Fe- Q15517 7 ( O75251 ) in wounded cells ; and ATPase , H+/K+ exchanging , beta polypeptide ( P51164 ) , and DB00171 synthase , H+ transporting , mitochondrial Fo complex , subunit P06681 ( subunit 9 ) ( Q06055 ) in ischemic cells . CONCLUSIONS : LILI at 660 nm stimulates the upregulation of genes coding for subunits of enzymes involved in complexes I and IV and DB00171 synthase . Construction of network for protein kinases that play a role in acute pancreatitis . OBJECTIVES : This study aimed to search for protein kinases that play a role in acute pancreatitis and analyze their potential connection with each other . METHODS : Information of human protein kinases were collected in protein kinase database , and then a systematic search was performed using PubMed for studies addressing the association between these kinases and acute pancreatitis . Gene Ontology Annotations were used to build interactions network for acute pancreatitis-associated protein kinases . RESULTS : A total of 570 human protein kinases were found , in which 28 kinases play a role in acute pancreatitis . Among the 28 kinases , Q13546 , O60674 , P12931 , P00533 , P06241 , MET , P23458 , P29597 , and P42345 were annotated in Gene Ontology database . A gene ontology interactions network was built to visualize the common biological process these kinases participated in . CONCLUSIONS : This study provides observations that protein kinases participate in all the sequential events in the exocrine pancreas in acute pancreatitis and that protein kinases are potential therapeutical target for acute pancreatitis .

MH_train_1192

MH_train_1192

MH_train_1192

interacts_with DB08827?

multiple_choice

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 . Does increased leukotriene B4 in type 1 diabetes result from elevated cholesteryl ester transfer protein activity ? Elevated cholesteryl ester transfer protein ( P11597 ) activity has been reported in type 1 diabetic subjects and may be one cause of the high incidence of macrovascular complications in these patients . LDL delivers arachidonic acid ( AA ) , in the form of cholesteryl ester ( CE ) , to cells such as monocytes and fibroblasts , as precursor for eicosanoid synthesis . We discovered that AA content in LDL CE was significantly correlated with P11597 activity , even after controlling for P11597 concentration , in type 1 diabetic children . The production of Q06643 (4) , a potent chemotactic and pro-inflammatory factor which plays a role in atherogenesis , has been shown to be increased in type 1 diabetic patients . We hypothesized that in these subjects , increased AA content in LDL CE , resulting from increased P11597 activity and transient hyperinsulinemia , may lead to enhanced synthesis of Q06643 (4) and subsequently the higher incidence of cardiovascular disease . Ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in human aqueous humor . PURPOSE : The aim of this study was to evaluate the ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in subjects undergoing routine cataract surgery with intraocular lens implantation . METHODS : An open-label , phase II confirmatory study of 54 subjects undergoing cataract surgery with intraocular lens implantation . A single drop of bromfenac sodium ophthalmic solution 0.1 % was administered at 30 , 60 , 90 , 120 , 180 , or 240 min prior to the initiation of cataract surgery . Samples of aqueous humor were aspirated through a paracentesis and analyzed by using high-performance liquid chromatography . Based upon these data , predicted concentrations of bromfenac in the aqueous humor over 24 h were generated by using computer simulation and compared with the IC(50) for bromfenac as a measure of anti-inflammatory efficacy . RESULTS : Peak aqueous-humor concentrations of bromfenac occurred between 150 and 180 min following ophthalmic dosing , with a mean concentration of 78.7 ng/mL . The level of bromfenac decreased in a log-linear fashion with an elimination-rate constant of 1.4 . DB00963 remained above the IC(50) value of cyclo-oxygenase-2 ( P35354 ) during the evaluated time points and over the 12-h dosing interval , using a computer model of extrapolated time points and assuming first-order elimination . CONCLUSIONS : Pharmacokinetic modeling , based upon samples collected over 240 min after a single dose of bromfenac sodium ophthalmic solution 0.1 % suggests that aqueous-humor concentrations remain at clinically effective levels ( above its IC(50) value for P35354 ) for over 12 h . Based upon this rationale , these results supported clinical trials that demonstrated the efficacy of twice-daily dosing of bromfenac sodium ophthalmic solution 0.1 % to manage patients with postoperative ocular pain and inflammation . P11926 is a critical determinant of P04198 oncogenesis and a therapeutic target in neuroblastoma . Neuroblastoma is a frequently lethal childhood tumor in which MYC gene deregulation , commonly as P04198 amplification , portends poor outcome . Identifying the requisite biopathways downstream of MYC may provide therapeutic opportunities . We used transcriptome analyses to show that P04198 -amplified neuroblastomas have coordinately deregulated myriad polyamine enzymes ( including P11926 , P19623 , P52788 , P17707 , O95190 , and Q9NWM0 ) to enhance polyamine biosynthesis . High-risk tumors without P04198 amplification also overexpress P11926 , the rate-limiting enzyme in polyamine biosynthesis , when compared with lower-risk tumors , suggesting that this pathway may be pivotal . Indeed , elevated P11926 ( independent of P04198 amplification ) was associated with reduced survival in a large independent neuroblastoma cohort . As polyamines are essential for cell survival and linked to cancer progression , we studied polyamine antagonism to test for metabolic dependence on this pathway in neuroblastoma . The Odc inhibitor alpha-difluoromethylornithine ( DB06243 ) inhibited neuroblast proliferation in vitro and suppressed oncogenesis in vivo . DB06243 treatment of neuroblastoma-prone genetically engineered mice ( TH- P04198 ) extended tumor latency and survival in homozygous mice and prevented oncogenesis in hemizygous mice . In the latter , transient Odc ablation permanently prevented tumor onset consistent with a time-limited window for embryonal tumor initiation . Importantly , we show that DB06243 augments antitumor efficacy of conventional cytotoxics in vivo . This work implicates polyamine biosynthesis as an arbiter of P04198 oncogenesis and shows initial efficacy for polyamine depletion strategies in neuroblastoma , a strategy that may have utility for this and other MYC-driven embryonal tumors . 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 . Bilirubin measured on a blood gas analyser : a suitable alternative for near-patient assessment of neonatal jaundice ? The reliability of a recently released total bilirubin assay for a blood gas analyser was assessed in two Australian hospital laboratories . The instrument computes total bilirubin concentration from multi-wavelength absorbance measurements of undiluted whole blood or plasma . Performance of the Radiometer P00519 735 blood gas analyser bilirubin method ( software version 3.6 ) was compared with a proven Roche diazo method for Hitachi analysers , calibrated using primary standards prepared from NIST P19623 916a bilirubin . Acceptable bilirubin results were found over a wide concentration range for most neonatal samples of whole blood or plasma . For adult specimens , bilirubin results were approximately 10 % lower on the blood gas analyser . Within-run imprecision ( whole blood ) was < 2.5 % , between-day imprecision ( synthetic controls ) < 1.0 % , and the bilirubin assay for both whole blood and plasma was linear to 1,000 micromol/L . Using sampling options from 35 microL to 195 microL , bilirubin results differed by less than 3 % , with a 95 microL syringe option producing the highest results . We conclude that the Radiometer P00519 735 bilirubin assay is suitable for near-patient assessment of neonatal jaundice using whole blood , thus eliminating the need for sample centrifugation . Verification using laboratory methods can be used when required . A positive correction of approximately 10 % is required for adult specimens to conform with Hitachi results ( P19623 916a calibration ) , possibly due to the optical characteristics of the higher proportion of conjugated bilirubin and other substances present in most adult samples . DB04690 enhances random integration of transfected DNA into the genome of mammalian cells . In order to study the involvement of P11387 ( top1 ) in recombination , we examined the effect of the anti-neoplastic drug camptothecin , which selectively poisons top1 by trapping top1-cleavable complexes on integration of exogenic vector into the genome of mammalian cells . We transfected mouse P00740 teratocarcinoma cells as well as Chinese hamster V79 cells with a plasmid carrying a selectable neo gene treated with camptothecin , and determined the frequency of neo+ ( G418(R) ) colonies . We found that treatment with camptothecin for as short a time as 4 h after electroporation resulted in a 4- to 33-fold stimulation of plasmid integration into the recipient genome via non-homologous recombination . These results imply that top1-cleavable complexes trapped by camptothecin could be potentially recombinogenic structures and could stimulate non-homologous recombination in vivo , promoting the integration of transfected plasmids into mammalian genome . 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 . Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 receptor ( IL13Rα2 ) that differs from the physiological P24394 /IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL-13-PE ) encoding a mutated human P35225 fused to Pseudomonas exotoxin ( mhIL-13-PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 that binds to the physiological P24394 /IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL-13-PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL-13-PE and two protein formulations , hIL-13-PE used in clinical trials ( DB05305 ) and a second-generation mhIL-13-PE . DB05305 doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL-13-PE led to ∼40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL-13-PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 include its short half-life , which demanded frequent or continued administration , and binding to P24394 /IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM . 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 . Direct and irreversible inhibition of cyclooxygenase-1 by nitroaspirin ( DB05822 ) . Benzoic acid , 2-(acetyl-oxy)-3-[(nitrooxy)methyl]phenyl ester ( DB05822 ) , a new drug made by an aspirin molecule linked , through a spacer , to a nitric oxide ( NO ) -donating moiety , is now under clinical testing for the treatment of atherothrombotic conditions . DB00945 exerts its antithrombotic activity by irreversibly inactivating platelet cyclooxygenase ( P36551 ) -1 . DB05822 in vivo undergoes metabolism into deacetylated and/or denitrated metabolites , and it is not known whether DB05822 needs to liberate aspirin to inhibit P23219 , or whether it can block it as a whole molecule . The aim of our study was to evaluate the effects of DB05822 and its analog or metabolites on platelet P23219 and whole blood P35354 and on purified ovine P36551 ( oCOX ) -1 and oCOX-2 . In particular , we have compared the mechanism by which DB05822 inhibits purified oCOX enzymes with that of aspirin using a spectrophotometric assay . All the DB05822 derivatives containing acetylsalicylic acid inhibited the activity of oCOX-1 and oCOX-2 , whereas the deacetylated metabolites and the nitric oxide-donating moiety were inactive . Dialysis experiments showed that oCOX-1 inhibition by DB05822 , similar to aspirin , is irreversible . Reversible P36551 inhibitors ( indomethacin ) or salicylic acid incubated with the enzyme before DB05822 prevent the irreversible inhibition of oCOX-1 by DB05822 as well as by aspirin . In conclusion , our data show that DB05822 acts as a direct and irreversible inhibitor of P23219 and that the presence of a spacer and NO-donating moiety in the molecule slows the kinetics of P23219 inhibition by DB05822 , compared with aspirin . 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 . Expression of nerve growth factor and matrix metallopeptidase-9/tissue inhibitor of metalloproteinase-1 in asthmatic patients . OBJECTIVE : The aim of this study was to measure the level of nerve growth factor ( P01138 ) in bronchial specimens from humans and to determine whether it correlated with not only clinical characteristics of asthma such as percent eosinophils , Th2 cytokine levels , and pulmonary function , but also metallopeptidase-9 ( P14780 ) and tissue inhibitor of metalloproteinases-1 ( P01033 ) . METHODS : Fifty-three people participated ; 42 had asthma . The participants underwent bronchoscopy and the specimens were analyzed . The participants ' clinical data including pulmonary function tests were reviewed . RESULTS : Bronchoalveolar lavage fluid ( BALF ) from patients with asthma had a significantly higher level of P01138 compared with that from participants without asthma . P01138 level showed a positive correlation with the percentage of eosinophils in both BALF and serum . The concentration of P01138 did not correlate with that of Th2 cytokines interleukin ( IL ) -4 , P05113 , and P35225 in BALF or parameters of pulmonary function including degree of airway hyperresponsiveness ( Q5SW96 ) . The levels of P14780 and P01033 in BALF were higher in asthma patients than in participants without asthma . The levels of P01138 correlated with P01033 levels but not with P14780 in the whole participants . CONCLUSIONS : This study shows that P01138 correlates with levels of eosinophils , a major effector cell in asthma . The high expression of P01138 and P01033 in asthma patients and the moderate correlation between P01138 and P01033 in the entire group of asthma subjects suggest a possible association between P01138 and P01033 , which may influence asthma pathogenesis . 5- DB00233 Inhibits Acute Clostridium difficile Toxin A-Induced Colitis in Rats . We tested the hypothesis that DB00244 ( DB00244 ) inhibits toxin A-induced generation of colonic leukotriene B4 ( LTB4 ) and toxin A colitis in rats . Isolated colonic segments in anesthetized rats were treated intraluminally with toxin A for 3 hours with or without 30 minutes of pretreatment with either DB00244 or sulfapyridine and then colonic tissue levels of LTB4 were measured and inflammation was assessed . Separately , sulfasalazine was administered to rats in their drinking water for 5 days , isolated colonic segments were then prepared , toxin A was administered , and inflammation was assessed as before . Pretreatment with DB00244 inhibited toxin A-induced increased tissue LTB4 concentration in the colon . Sulfasalazine and DB00244 but not sulfapyridine significantly inhibited toxin A colitis . However , pretreatment with DB00244 did not protect against direct Q8NER1 -mediated colitis caused by capsaicin . Toxin A stimulated the release of DB05875 ( SP ) , and this effect was also inhibited by sulfasalazine and DB00244 but not by sulfapyridine . Thus , toxin A stimulates colonic LTB4 resulting in activation of Q8NER1 , release of SP , and colitis . Inhibition of P09917 by DB00244 disrupts this pathway and supports the concept that LTB4 activation of Q8NER1 plays a role in toxin A colitis . Elevated expression of P07237 family proteins during differentiation of mouse P00740 teratocarcinoma cells . We investigated the expression of protein disulfide isomerase family proteins ( P07237 , ERp61 , and P13667 ) in mouse P00740 teratocarcinoma cells during differentiation induced by treatment with retinoic acid and dibutyryl DB02527 . Each member of this family was expressed at a constitutive level in undifferentiated P00740 cells . During differentiation of P00740 cells to parietal or visceral endodermal cells the protein level of all these enzymes increased , although the extent of this increase in both protein and mRNA levels varied among the enzymes . Certain proteins were found to be coimmunoprecipitated with P07237 , ERp61 , and P13667 in the presence of a chemical crosslinker . Type IV collagen was significantly coprecipitated with P07237 whereas laminin was equally coprecipitated with the three proteins . Furthermore , 210 kDa protein characteristically coprecipitated with P13667 . Thus , the induction of P07237 family proteins during the differentiation of P00740 cells and their association with different proteins may implicate specific functions of each member of this family despite the common redox activity capable of catalyzing the disulfide bond formation . The adenine nucleotide translocator 1 acts as a type 2 transglutaminase substrate : implications for mitochondrial-dependent apoptosis . In this study we provide in vitro and in vivo evidence showing that the protein disulphide isomerase ( P07237 ) activity of type 2 transglutaminase ( TG2 ) regulates the correct assembly and function of the mitochondrial ADP/ DB00171 transporter adenine nucleotide translocator 1 ( P12235 ) . We demonstrate , by means of biochemical and morphological analyses , that P12235 and TG2 physically interact in the mitochondria . Under physiological conditions , TG2 's P07237 activity regulates the ADP/ DB00171 transporter function by controlling the oligomerization of P12235 . In fact , mitochondria isolated from hearts of TG2(-/-) mice exhibit increased polymerization of P12235 , paralleled by an enhanced ADP/ DB00171 carrier activity , as compared to mitochondria belonging to TG2(+/+) mice . Interestingly , upon cell-death induction , P12235 becomes a substrate for TG2 's cross-linking activity and the lack of TG2 results in a reduction of apoptosis as well as in a marked sensitivity to the ADP/ DB00171 exchange inhibition by atractyloside . These findings suggest a complex TG2-dependent regulation of the ADP/ DB00171 transporter and reveal new important avenues for its potential applications in the treatment of some mitochondrial-dependent diseases , including cardiovascular and neurodegenerative diseases . Pharmacological characterization of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel selective P09917 -activating protein inhibitor that reduces acute and chronic inflammation . Leukotrienes ( LTs ) are proinflammatory lipid mediators synthesized by the conversion of arachidonic acid ( AA ) to P01374 (4) by the enzyme P09917 ( P09917 ) in the presence of P09917 -activating protein ( P20292 ) . 3-[3-tert-Butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) is a novel selective P20292 inhibitor in development for the treatment of respiratory conditions such as asthma . In a rat ex vivo whole-blood calcium ionophore-induced Q06643 (4) assay , DB05225 ( administered orally at 1 mg/kg ) displayed > 50 % inhibition for up to 6 h with a calculated EC(50) of approximately 60 nM . When rat lung was challenged in vivo with calcium ionophore , DB05225 inhibited Q06643 (4) and cysteinyl leukotriene ( CysLT ) production with ED(50) values of 0.8 and 1 mg/kg , respectively . In this model , the EC(50) derived from plasma DB05225 was approximately 330 nM for inhibition of both Q06643 (4) and CysLT . In an acute inflammation setting , DB05225 displayed dose-dependent inhibition of Q06643 (4) , CysLT , and plasma protein extravasation induced by peritoneal zymosan injection . In a model of chronic lung inflammation using ovalbumin-primed and challenged BALB/c mice , DB05225 reduced the concentrations of eosinophil peroxidase , CysLTs , and interleukin-5 in the bronchoalveolar lavage fluid . Finally , DB05225 increased survival time in mice exposed to a lethal intravenous injection of platelet-activating factor . In summary , DB05225 is a novel , potent and selective P20292 inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute and chronic inflammation and in a model of lethal shock . Splenic autonomic denervation increases inflammatory status but does not aggravate atherosclerotic lesion development . The brain plays a prominent role in the regulation of inflammation . Immune cells are under control of the so-called cholinergic anti-inflammatory reflex , mainly acting via autonomic innervation of the spleen . Activation of this reflex inhibits the secretion of proinflammatory cytokines and may reduce the development of atherosclerosis . Therefore , the aim of this study was to evaluate the effects of selective parasympathetic ( Px ) and sympathetic ( Sx ) denervation of the spleen on inflammatory status and atherosclerotic lesion development . Female P02649 *3-Leiden. P11597 mice , a well-established model for human-like lipid metabolism and atherosclerosis , were fed a cholesterol-containing Western-type diet for 4 wk after which they were subdivided into three groups receiving either splenic Px , splenic Sx , or sham surgery . The mice were subsequently challenged with the same diet for an additional 15 wk . Selective Px increased leukocyte counts ( i.e. , dendritic cells , B cells , and T cells ) in the spleen and increased gene expression of proinflammatory cytokines in the liver and peritoneal leukocytes compared with Sx and sham surgery . Both Px and Sx increased circulating proinflammatory cytokines IL-1β and P05231 . However , the increased proinflammatory status in denervated mice did not affect atherosclerotic lesion size or lesion composition . CONCLUSION : Predominantly selective Px of the spleen enhances the inflammatory status , which , however , does not aggravate diet-induced atherosclerotic lesion development . 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 . 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 . DB02426 effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 and P05141 may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 -/- mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 -independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 -/- brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 -independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 -/- mice . However , in liver , only Ant2 mRNA was found , whereas in brown adipose tissue , Ant1 and Ant2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 isoform mediates fatty-acid-induced uncoupling , whereas the P12235 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria . 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 .

MH_train_1193

MH_train_1193

MH_train_1193

interacts_with DB08907?

multiple_choice

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 . Identification of verrucarin a as a potent and selective steroid receptor coactivator-3 small molecule inhibitor . Members of the steroid receptor coactivator ( P12931 ) family are overexpressed in numerous types of cancers . In particular , steroid receptor coactivator 3 ( Q9Y6Q9 ) has been recognized as a critical coactivator associated with tumor initiation , progression , recurrence , metastasis , and chemoresistance where it interacts with multiple nuclear receptors and other transcription factors to enhance their transcriptional activities and facilitate cross-talk between pathways that stimulate cancer progression . Because of its central role as an integrator of growth signaling pathways , development of small molecule inhibitors ( SMIs ) against SRCs have the potential to simultaneously disrupt multiple signal transduction networks and transcription factors involved in tumor progression . Here , high-throughput screening was performed to identify compounds able to inhibit the intrinsic transcriptional activities of the three members of the P12931 family . Verrucarin A was identified as a SMI that can selectively promote the degradation of the Q9Y6Q9 protein , while affecting Q15788 and P12931 -2 to a lesser extent and having no impact on CARM-1 and p300 protein levels . Verrucarin A was cytotoxic toward multiple types of cancer cells at low nanomolar concentrations , but not toward normal liver cells . Moreover , verrucarin A was able to inhibit expression of the Q9Y6Q9 target genes P08253 and P45452 and attenuated cancer cell migration . We found that verrucarin A effectively sensitized cancer cells to treatment with other anti-cancer drugs . Binding studies revealed that verrucarin A does not bind directly to Q9Y6Q9 , suggesting that it inhibits Q9Y6Q9 through its interaction with an upstream effector . In conclusion , unlike other P12931 SMIs characterized by our laboratory that directly bind to SRCs , verrucarin A is a potent and selective SMI that blocks Q9Y6Q9 function through an indirect mechanism . 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) . A set of consensus mammalian mediator subunits identified by multidimensional protein identification technology . The Mediator is a multiprotein transcriptional coactivator that is expressed ubiquitously in eukaryotes from yeast to mammals and is required for induction of RNA polymerase II ( pol II ) transcription by DNA binding transcription factors . In the work described here , we exploit multidimensional protein identification technology ( MudPIT ) to carry out a proteomic analysis of the subunit composition of the mammalian Mediator complex . By comparing MudPIT data sets obtained from six independent Mediator preparations immunoaffinity purified through their Nut2 ( Q9BTT4 ) , Med25 ( Q9NWA0 ) , Intersex ( Q9NX70 ) , A0JLT2 ( A0JLT2 ) , AK007855 ( Q9H204 ) , or CRSP70 ( O95402 ) subunits , we identify a set of consensus mammalian Mediator subunits . In addition , we identify as Mediator-associated proteins the P49336 -like cyclin-dependent kinase CDK11 and the Q9UHV7 -like Q71F56 protein ( Q71F56 ) , which is mutated in patients with the congenital heart defect transposition of the great arteries ( TGA ) . DB06693 reduces cartilage degradation in rabbit experimental osteoarthritis through inhibition of synovial inflammation . OBJECTIVE : To examine the therapeutic efficacy of an P04035 inhibitor ( statin ) in rabbit osteoarthritis ( OA ) in vitro and in vivo . METHODS : In the presence or absence of mevastatin , rabbit chondrocytes and synoviocytes were incubated with Interleukin-1beta ( IL-1beta ) , and analyzed by biochemical methods . Thirty-two mature rabbits that underwent bilateral anterior cruciate ligament transaction ( ACLT ) received six consecutive weekly intra-articular injections of mevastatin at three different concentrations or a control solution . All animals were sacrificed 6 weeks after ACLT , and the knee joints were assessed by morphological , histological , immunohistochemical , and biochemical methods . RESULTS : DB06693 inhibited IL-1beta stimulation of gene expression of monocyte chemoattractant protein-1 ( P13500 ) and matrix-metalloproteinases 3 ( P08254 ) , in synoviocytes but not chondrocytes . The levels of P13500 and P08254 productions in synoviocytes were significantly reduced by statin-treatment . In rabbit with OA , intra-articular injection of mevastatin significantly reduced cartilage degradation , as assessed by morphological and histological examinations . Synovial tissues of knees treated with mevastatin showed less severe inflammatory responses with reduced thickness of synovial cell lining and less infiltration of subsynovial P34810 +monocyte lineage cells compared to untreated control knees . Relative mRNA expressions of P13500 , IL-1beta , P08254 , and P45452 were reduced in synovial tissues , but not articular cartilage , of knees treated with mevastatin compared with untreated control knees . CONCLUSION : During the development of experimental OA , intra-articular administration of P04035 inhibitor ( statin ) reduces inflammatory cell infiltration and matrix-degrading enzyme expression , thus limiting cartilage degradation . 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 . A phase II study of DB05434 ( thrombospondin-1 analog ) for the treatment of metastatic melanoma . OBJECTIVES : Thrombospondins are natural inhibitors of angiogenesis , tumor metastases , and tumor growth ( melanoma ) . DB05434 is a synthetic analog of thrombospondin-1 , well tolerated in phase I studies . We conducted a phase II trial evaluating the clinical efficacy of DB05434 and its effects on biomarkers of angiogenesis and immunity in patients with metastatic melanoma ( MM ) . PATIENTS AND METHODS : A 2-stage phase II clinical trial was conducted to assess the clinical efficacy , safety , and pharmacodynamic effects ( angiogenesis and immunity ) of DB05434 in patients with stage IV melanoma . The primary endpoint was 18-week treatment failure rate . Patients self-administered 100 mg of DB05434 subcutaneously twice daily . Blood samples were collected at baseline and every 3 weeks while on therapy . Eligible patients demonstrated measurable disease , good performance status and no evidence of intracranial metastases . Correlative laboratory studies evaluated biomarkers of angiogenesis and immunity . RESULTS : Twenty-one patients were enrolled . Most patients were stage M1c ( 71 % ) and all had prior therapy for MM . Only 3 of the first 20 patients enrolled were progression free and on treatment at 18 weeks resulting in early termination of the study . Decreases in peripheral blood P15692 levels and P49767 levels , and CD146 and P28906 /133 counts relative to pretreatment were detected . Limited changes in antitumor T cell immunity were observed . CONCLUSIONS : DB05434 therapy administered at 100 mg twice/day in patients with MM did not demonstrate definite clinical efficacy . Further dose escalation or combination with cytotoxic therapy may be more effective therapeutically . DB08810 protects against ethanol-induced gastric mucosal injury in rats : role of 5-hydroxytryptamine , prostaglandins and sulfhydryl compounds . This study was designed to determine the gastroprotective properties of cinitapride ( CNT ) , a novel prokinetic benzamide derivative agonist of Q13639 and 5-HT1 receptors and 5-HT2 antagonist , on mucosal injury produced by 50 % ( v/v ) ethanol . Results were compared with those for 5-hydroxytryptamine ( 5-HT : 10 mg kg-1 ) . The possible involvements of gastric mucus secretion , endogenous prostaglandins ( PGs ) and sulfhydryl compounds ( SH ) in the protection mediated by CNT were also examined . Intraperitoneal administration of CNT ( 0.50 and 1 mg kg-1 ) , 30 min before ethanol , significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus . CNT ( 1 mg kg-1 ) also produced a significant increase in gastric mucosal levels of DB00917 , but did not induce any significant changes in SH values . On the contrary , pretreatment with 5-HT worsened ethanol-induced erosions , however , did not affect gastric mucus secretion , glycoprotein content or DB00917 levels , although the non-protein SH fraction was significantly decreased . The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism . We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved . 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 . Repeated administration of a F(ab')2 fragment of an anti-tumor necrosis factor alpha monoclonal antibody in patients with severe sepsis : effects on the cardiovascular system and cytokine levels . In an uncontrolled clinical trial the effects of repeated administration of the F(ab')2 fragment of a murine monoclonal anti-tumor necrosis factor alpha ( P01375 alpha ) -antibody ( DB04956 ) on cytokine levels and the cardiovascular system were studied in 20 patients with severe sepsis . Patients were treated with a total of 11 single dosages of the anti- P01375 alpha-antibody intravenously over 5 days using either 1 mg/kg ( n = 10 ) or 3 mg/kg ( n = 10 ) . The anti- P01375 alpha-antibody was well tolerated in all patients without signs of toxicity and without development of anti-murine antibodies . As assessed by cytokine levels ( P01375 alpha , P05231 ) and hemodynamics there was no evidence that the higher dosage of the anti- P01375 alpha-antibody ( 3 mg/kg per dose ) was more effective than the lower dosage ( 1 mg/kg per dose ) . Comparison of our data with recent data from phase I or II trials using a complete murine monoclonal anti- P01375 alpha-antibody suggest that the F(ab')2 fragments of the murine monoclonal anti- P01375 alpha-antibody may be of similar efficacy . Definitive conclusions , however , with respect to improvement of mortality and improvement of the cardiovascular system , await the results of larger ongoing placebo-controlled trials . Estradiol increases cell growth in human astrocytoma cell lines through ERα activation and its interaction with Q15788 and Q9Y6Q9 coactivators . Estradiol ( E2 ) regulates several cellular functions through the interaction with estrogen receptor subtypes , ERα and ERβ , which present different functional and regulation properties . ER subtypes have been identified in human astrocytomas , the most common and aggressive primary brain tumors . We studied the role of ER subtypes in cell growth of two human astrocytoma cell lines derived from tumors of different evolution grades : U373 and D54 ( grades III and IV , respectively ) . E2 significantly increased the number of cells in both lines and the co-administration with an ER antagonist ( ICI 182 , 780 ) significantly blocked E2 effects . ERα was the predominant subtype in both cell lines . E2 and ICI 182 , 780 down-regulated ERα expression . The number of U373 and D54 cells significantly increased after PPT ( ERα agonist ) treatment but not after DPN ( ERβ agonist ) one . To determine the role of Q15788 and Q9Y6Q9 coactivators in ERα induced cell growth , we silenced them with RNA interference . Coactivator silencing blocked the increase in cell number induced by PPT . The content of proteins involved in proliferation and metastasis was also determined after PPT treatment . Western blot analysis showed that in U373 cells the content of PR isoforms ( PR-A and PR-B ) , P00533 , P15692 and cyclin D1 increased after PPT treatment while in D54 cells only the content of P00533 was increased . Our results demonstrate that E2 induces cell growth of human astrocytoma cell lines through ERα and its interaction with Q15788 and Q9Y6Q9 and also suggest differential roles of ERα on cell growth depending on astrocytoma grade . Genetics of bipolar disorder . Many linkage loci and candidate genes have been reported in molecular genetic studies of bipolar disorder . However , none of these findings have been consistently replicated . Meta-analyses of linkage studies have also reported conflicting results . Among recently reported candidate genes , P23560 , P59103 , P31749 , Q12879 , P17861 , P35626 , Q13639 , O14732 and P14867 may have some importance . Study of the possible roles of epigenetics or analysis of genetic diseases , in which bipolar disorder is one of phenotypes , may also be promising . In addition to monoaminergic and intracellular signaling pathways , recent studies have revealed possible roles for mitochondrial dysfunction , for glutamatergic dysfunction and for the endoplasmic reticulum stress pathway . F-actin involvement in guinea pig sperm motility . Sperm motility is a must for natural fertilization to occur . During their travel through the epididymis , mammalian spermatozoa gradually acquire the ability to move . This is accomplished through a sliding movement of the outer doublet microtubules of the axoneme which is energized by the dynein ATPase . Within its complex structure , the mammalian sperm flagellum contains F-actin and thus , we decided to test in the guinea pig sperm flagellum the role of F-actin in motility . During maturation , capacitation , and the acrosome reaction , a gradual decrease of the relative concentration of F-actin was observed . Motility increased as spermatozoa became able to fertilize . P06396 , phalloidin , and KI inhibited sperm motility . P06396 canceled sperm motility within 20 min of treatment while 0.6 M KI had immediate effects . Phalloidin diminished hyperactive sperm motility slightly . All three compounds significantly increased the relative concentration of F-actin . Latrunculins are conventional drugs that destabilize the F-actin cytoskeleton . DB02621 ( O43561 A ) did not affect sperm motility ; but significantly increased F-actin relative concentration . The results suggested that in guinea pig spermatozoa , randomly severing F-actin filaments inhibits flagellar motility ; while end filament alteration does not . Thus , specific filament regions seem to be important for sperm motility . Drugs targeting nitric oxide synthase for migraine treatment . INTRODUCTION : Ample evidence that nitric oxide ( NO ) is a causative molecule in migraine has encouraged research to develop drugs that target the NO-cGMP cascade for migraine treatment . NO synthase ( NOS ) inhibition is an innovative therapeutic principle . AREAS COVERED : This paper reviews the rationale underlying NOS inhibition in migraine treatment . It also provides a review on the efficacy and safety data for NOS inhibitors ( nonselective NOS inhibitor L-N(G)-methyl-arginine hydrochloride [ L-NMMA ] , selective inducible NOS [ P35228 ] inhibitors GW273629 and GW274150 , combined neuronal NOS [ P29475 ] inhibitor and P28222 /1D receptor agonist DB06096 ) in acute or preventive migraine treatment . EXPERT OPINION : The data highlighted herein , from four placebo-controlled trials and 1 open-labeled clinical trial using 4 different NOS inhibitors on a total of 705 patients , provide convincing efficacy data only for the nonselective NOS inhibitor L-NMMA . Unfortunately , this NOS inhibitor raises cardiovascular safety concerns and has an unfavorable pharmacokinetic profile . As experimental studies predicted , P35228 inhibitors are ineffective in migraine . Still , upcoming selective P29475 inhibitors are a hope for migraine treatment , with the P29475 isoform being most clearly involved in trigeminovascular transmission and central sensitization . Future studies should help to clarify whether NOS inhibition is equally fruitful in acute and preventive treatment . It should also clarify if P29475 inhibition holds promise as a therapeutic tool for the treatment of chronic migraine and other forms of headache . Crosstalk between circulating peroxisome proliferator-activated receptor gamma , adipokines and metabolic syndrome in obese subjects . BACKGROUND : P37231 ( PPARγ ) has direct and indirect function in adipokines production process . We aimed to assess the possible influence of circulating PPARγ on relative risk of metabolic syndrome and also examine the association between circulating PPARγ and adipokines levels among obese subjects . METHODS : A total of 96 obese subjects ( body mass index ( BMI ) ≥30 ) were included in the current cross-sectional study . We assessed the body composition with the use of Body Composition Analyzer BC-418MA - Tanita . The MetS ( metabolic syndrome ) was defined based on the National DB04540 Education Program Adult Treatment Panel III . All baseline blood samples were obtained following an overnight fasting . Serum concentrations of adipokines including DB00162 binding protein 4 ( P02753 ) , omentin-1 , vaspin , progranulin , nesfatin-1 and circulating PPARγ was measured with the use of an enzyme-linked immunosorbent assay method . Statistical analyses were performed using software package used for statistical analysis ( SPSS ) . RESULTS : We found main association between circulating PPARγ and body composition in obese population . The risk of metabolic syndrome in subjects with higher concentration of PPARγ was 1.9 fold in compared with lower concentration of PPARγ after adjustment for age , sex and BMI . There was significant association between PPARγ and adipokines , specially nesfatin-1 and progranulin . Defined adipokines pattern among participants demonstrated the markedly higher concentration of vaspin , P02753 and nesfatin-1 in participants with MetS compared to non-MetS subjects . CONCLUSIONS : It appears all of studied adipokines might have association with PPARγ level and might simultaneously be involve in some common pathway to make susceptible obese subjects for MetS . Tumor-suppressive microRNA-497 targets IKKβ to regulate NF-κB signaling pathway in human prostate cancer cells . BACKGROUND : Prostate cancer ( PCa ) is one of the most prevalent malignant tumors , PCa-related death is mainly due to the high probability of metastasis . MicroRNAs ( miRNAs ) play an important role in cancer initiation , progression and metastasis by regulating their target genes . METHODS : real-time PCR was used to detected the expression of microRNA-497 . The molecular biological function was investigated by using cell proliferation assays , cell cycle assay , and migration and invasion assay . We used several Algorithms and confirmed that IKKβ is directly regulated by miR-497 . RESULTS : Here , we found miR-497 is downregulated in human prostate cancer ( PCa ) and inhibites the proliferation activity , migration and invasion of PC3-AR cells . Subsequently , IKKβ is confi rmed as a target of miR-497 . Furthermore , knockdown of IKKβ expression resulted in decreased proliferation activity , migration and invasion . Finally , similar results was found after treatment with a novel IKK-β inhibitor ( IMD-0354 ) in PC3-AR cells . P49336 , P14780 , and PSA were involved in all these process . CONCLUSION : Taken together , our results show evidence that miR-497 may function as a tumor suppressor genes by regulating IKK-β in PCa , and may provide a strategy for blocking PCa metastasis . Differences in hepatotoxicity and gene expression profiles by anti-diabetic Q07869 gamma agonists on rat primary hepatocytes and human HepG2 cells . Agonists of peroxisome proliferator-activated receptor gamma ( PPARgamma ) are a new class of oral drugs designed to treat insulin-resistant diabetes ( i.e. , type 2 diabetes ) . However , troglitazone , the first compound in the class approved by the US Food and Drug Administration ( FDA ) in 1997 was found to be hepatotoxic and was withdrawn from the market after reports of severe liver failure . The mechanism of Q07869 gamma agonist-induced hepatotoxicity remains unknown . In this study , we examined the hepatotoxic effects of five Q07869 gamma agonists ( ciglitazone , pioglitazone , rosiglitazone , troglitazone , and DB04971 ) on rat primary hepatocytes and human HepG2 cells . We also compared the gene expression profiles of rat primary hepatocytes after exposure to Q07869 gamma agonists by using the Rat Genome Survey Microarray system from Applied Biosystems in order to understand the mechanisms of hepatotoxicities induced by PPARgamma agonists . Consistent with the hepatotoxicity data , our results demonstrate that the gene expression profiles affected by troglitazone and ciglitazone can be clearly distinguished from those by pioglitazone and rosiglitazone . Genes that are differentially expressed between the more toxic troglitazone/ciglitazone group and the less toxic rosiglitazone/pioglitazone group are involved in necrotic , apoptotic , and cell proliferative pathways . The five compounds were also clustered based on a set of molecular descriptors . The clustering based on chemical structural information is in good agreement with the clustering of compounds based on cytotoxicity or gene expression data , indicating a strong relationship between chemical structure and biological endpoints . Our work suggests that microarray analysis together with toxicological observations can be used to rank drugs for hepatotoxicity and to evaluate the safety of new compounds . Q9UH65 regulates mast cell FcepsilonRI-mediated signaling and anaphylaxis . Mast cells , perhaps best known by their ability to trigger allergic reactions after stimulation through the FcepsilonRI , express the unusual phosphatidylinositol 3-kinase ( PI3K ) -dependent , Rac-binding protein Q9UH65 . Here , we show that the IgE-mediated passive cutaneous and the systemic anaphylactic responses are strongly reduced in Q9UH65 (-/-) mice . Cultured Q9UH65 (-/-) immature bone marrow mast cells ( BMMC ) are also impaired in FcepsilonRI-mediated degranulation , which can be restored by expression of exogenous wild-type Q9UH65 , but less so if a phosphatidylinositol trisphosphate ( PIP(3) ) binding mutant is expressed . Q9UH65 itself supports inositol-3-phosphate and PIP(3) production , the latter indicating a potential feedback from Q9UH65 towards PI3K . FcepsilonRI-stimulated transcription and release of cytokines is controlled by Q9UH65 . Key FcepsilonRI signal transduction events like activation of O43561 by phosphorylation , activation of Akt/ P31749 and of p38 Q96HU1 kinase are reduced in Q9UH65 (-/-) BMMC , but P29323 is strongly hyperactivated . Some requirements for Q9UH65 were apparent only under limited-strength signaling conditions . We suggest that Q9UH65 defines a new element of efficient mast cell activation upon FcepsilonRI signaling , important for the control of mast cell-dependent anaphylaxis . [ 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 . Coactivators for the orphan nuclear receptor RORalpha . A mutation in the nuclear orphan receptor RORalpha results in a severe impairment of cerebellar development by unknown mechanisms . We have shown previously that RORalpha contains a strong constitutive activation domain in its C terminus . We therefore searched for mammalian RORalpha coactivators using the minimal activation domain as bait in a two-hybrid screen . Several known and putative coactivators were isolated , including glucocorticoid receptor-interacting protein-1 ( Q9Y3R0 ) and peroxisome proliferator-activated receptor ( Q07869 ) -binding protein ( PBP/ Q15648 / Q15648 ) . These interactions were confirmed in vitro and require the intact activation domain of RORalpha although different requirements for interaction with Q9Y3R0 and PBP were detected . Even in the absence of exogenous ligand , RORalpha interacts with a complex or complexes of endogenous proteins , similar to those that bind to ligand-occupied thyroid hormone and vitamin D receptors . Both PBP and Q9Y3R0 were shown to be present in these complexes . Thus we have identified several potential RORalpha coactivators that , in contrast to the interactions with hormone receptors , interact with RORalpha in yeast , in bacterial extracts , and in mammalian cells in vivo and in vitro in the absence of exogenous ligand . Q9Y3R0 functioned as a coactivator for the RORalpha both in yeast and in mammalian cells . Thus , Q9Y3R0 is the first proven coactivator for RORalpha . Quantitative liver-specific protein fingerprint in blood : a signature for hepatotoxicity . We discuss here a new approach to detecting hepatotoxicity by employing concentration changes of liver-specific blood proteins during disease progression . These proteins are capable of assessing the behaviors of their cognate liver biological networks for toxicity or disease perturbations . Blood biomarkers are highly desirable diagnostics as blood is easily accessible and baths virtually all organs . Fifteen liver-specific blood proteins were identified as markers of acetaminophen ( DB00316 ) -induced hepatotoxicity using three proteomic technologies : label-free antibody microarrays , quantitative immunoblotting , and targeted iTRAQ mass spectrometry . Liver-specific blood proteins produced a toxicity signature of eleven elevated and four attenuated blood protein levels . These blood protein perturbations begin to provide a systems view of key mechanistic features of DB00316 -induced liver injury relating to glutathione and DB00118 ( DB00118 ) depletion , mitochondrial dysfunction , and liver responses to the stress . Two markers , elevated membrane-bound catechol-O-methyltransferase ( MB- P21964 ) and attenuated retinol binding protein 4 ( P02753 ) , report hepatic injury significantly earlier than the current gold standard liver biomarker , alanine transaminase ( ALT ) . These biomarkers were perturbed prior to onset of irreversible liver injury . Ideal markers should be applicable for both rodent model studies and human clinical trials . Five of these mouse liver-specific blood markers had human orthologs that were also found to be responsive to human hepatotoxicity . This panel of liver-specific proteins has the potential to effectively identify the early toxicity onset , the nature and extent of liver injury and report on some of the DB00316 -perturbed liver networks . P04150 interacting protein-1 restores glucocorticoid responsiveness in steroid-resistant airway structural cells . Glucocorticoid ( GC ) insensitivity represents a profound challenge in managing patients with asthma . The mutual inhibition of transcriptional activity between GC receptor ( GR ) and other regulators is one of the mechanisms contributing to GC resistance in asthma . We recently reported that interferon regulatory factor ( Q969Q1 ) -1 is a novel transcription factor that promotes GC insensitivity in human airway smooth muscle ( P17405 ) cells by interfering with GR signaling ( Tliba et al. , Am J Respir Cell Mol Biol 2008;38:463-472 ) . Here , we sought to determine whether the inhibition of GR function by P10914 involves its interaction with the transcriptional co-regulator GR-interacting protein 1 ( Q9Y3R0 ) , a known GR transcriptional co-activator . We here found that siRNA-mediated Q9Y3R0 depletion attenuated P10914 -dependent transcription of the luciferase reporter construct and the mRNA expression of an P10914 -dependent gene , P28907 . In parallel experiments , Q9Y3R0 silencing significantly reduced GR-mediated transactivation activities . Co-immunoprecipitation and Q86UG4 pull-down assays showed that Q9Y3R0 , through its repression domain , physically interacts with P10914 identifying Q9Y3R0 as a bona fide transcriptional co-activator for P10914 . Interestingly , the previously reported inhibition of GR-mediated transactivation activities by either P01375 and P01579 treatment or P10914 overexpression was fully reversed by increasing cellular levels of Q9Y3R0 . Together , these data suggest that the cellular accumulation of P10914 may represent a potential molecular mechanism mediating altered cellular response to GC through the depletion of Q9Y3R0 from the GR transcriptional regulatory complexes .

MH_train_1194

MH_train_1194

MH_train_1194

interacts_with DB01259?

multiple_choice

Individuality in P05230 expression significantly influences platinum resistance and progression-free survival in ovarian cancer . BACKGROUND : Ovarian cancer is frequently advanced at presentation when treatment is rarely curative . Response to first-line platinum-based chemotherapy significantly influences survival , but clinical response is unpredictable and is frequently limited by the development of drug-resistant disease . METHODS : We used qRT-PCR analysis to assess intertumour differences in the expression of fibroblast growth factor 1 ( P05230 ) and additional candidate genes in human ovarian tumours ( n=187 ) , and correlated individuality in gene expression with tumour histology , chemotherapy response and survival . We used MTT assays to assess platinum chemosensitivity in drug-sensitive and drug-resistant ovarian cell lines . RESULTS : Marked intertumour differences in gene expression were observed , with each tumour having a unique gene expression profile . Nine genes , including P05230 ( P=1.7 × 10(-5) ) and P21802 ( P=0.003 ) , were differentially expressed in serous and nonserous tumours . Q00987 ( P=0.032 ) and P04626 ( P=0.064 ) expression was increased in platinum-sensitive patients , and P05230 ( adjusted log-rank test P=0.006 ) , P21802 ( P=0.04 ) and PDRFRB expression ( P=0.037 ) significantly inversely influenced progression-free survival . Stable P05230 gene knockdown in platinum-resistant A2780DPP cells re-sensitised cells to both cisplatin and carboplatin . CONCLUSION : We show for the first time that P05230 is differentially expressed in high-grade serous ovarian tumours , and that individuality in P05230 expression significantly influences progression-free survival and response to platinum-based chemotherapy . 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 . Targeted treatment of advanced and metastaticbreast cancer with lapatinib . Improved molecular understanding of breast cancer in recent years has led to the discovery of important drug targets such as HER-2 and P00533 . DB01259 is a potent dual inhibitor of HER-2 and P00533 . Preclinical and phase I studies have shown activity with lapatinib in a number of cancers , including breast cancer , and the drug is well tolerated . The main known drug interactions are with paclitaxel and irinotecan . The most significant side-effects of lapatinib are diarrhea and adverse skin events . Rates of cardiotoxicity compare favorably with trastuzumab , a monoclonal antibody against HER-2 . This paper focuses on lapatinib in advanced and metastatic breast cancer , which remains an important therapeutic challenge . Phase II and III studies show activity as monotherapy , and in combination with chemotherapy or hormonal agents . Results from these studies suggest that the main benefit from lapatinib is in the HER-2 positive breast cancer population . Combinations of lapatinib and trastuzumab are also being studied and show encouraging results , particularly in trastuzumab-refractory metastatic breast cancer . DB01259 may have a specific role in treating HER-2 positive CNS metastases . The role of lapatinib as neoadjuvant therapy and in early breast cancer is also being evaluated . A double-blind , placebo-controlled outpatient trial of pergolide for cocaine dependence . Results of preclinical studies suggest that pergolide , a mixed D(1)/ P14416 agonist , may be useful in treating cocaine dependence . To empirically investigate this possibility , we conducted a 5-year , double-blind , placebo-controlled clinical trial of two doses of pergolide ( 0.05 and 0.25 mg bid ) in subjects with cocaine dependence and combined cocaine/alcohol dependence . Data analysis was performed on an intent to treat population ( N=358 ) and a per protocol population ( N=108 ) with urine drug screens ( UDS ) used as the main outcome measure . There were no significant effects on UDS at either pergolide dose . DB01186 had no significant effect on alcohol use in the comorbid alcohol/cocaine dependence group . DB01186 does not appear to have clinical value in the treatment of cocaine dependence or in decreasing alcohol use in cocaine-dependent individuals at the presently studied doses . Dimerization effect of sucrose octasulfate on rat P05230 . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 are Lys113 , Lys118 , Arg122 and Lys128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 . This difference may be important for SOS-mediated P05230 dimerization in rat . Emerging small molecule drugs . Dyslipidaemia is a major risk factor for cardiovascular diseases . Pharmacological lowering of LDL-C levels using statins reduces cardiovascular risk . However , a substantial residual risk persists especially in patients with type 2 diabetes mellitus . Because of the inverse association observed in epidemiological studies of HDL-C with the risk for cardiovascular diseases , novel therapeutic strategies to raise HDL-C levels or improve HDL functionality are developed as complementary therapy for cardiovascular diseases . However , until now most therapies targeting HDL-C levels failed in clinical trials because of side effects or absence of clinical benefits . This chapter will highlight the emerging small molecules currently developed and tested in clinical trials to pharmacologically modulate HDL-C and functionality including new P11597 inhibitors ( anacetrapib , evacetrapib ) , novel Q07869 agonists ( K-877 , CER-002 , P15924 -8658 , INT131 and DB05187 ) , LXR agonists ( ATI-111 , LXR-623 , XL-652 ) and RVX-208 . Intraepithelial CD8-positive T lymphocytes predict survival for patients with serous stage III ovarian carcinomas : relevance of clonal selection of T lymphocytes . BACKGROUND : The aim of this study was to investigate the prognostic effect of tumour-infiltrating lymphocytes ( TILs ) in serous stage III ovarian carcinoma to determine Q15399 clonality and to correlate this to Her2/neu expression . METHODS : DB03843 -fixed and paraffin-embedded ovarian carcinomas were examined for P11836 - , CD3- , P01730 - and CD8-positive lymphocytes ( n=100 ) , and for Her2/neu-positive tumour cells ( n=55/100 ) by immunohistochemistry . Clonality analysis was carried out by T-cell receptor gamma ( TCRgamma ) gene rearrangements ( n=93/100 ) . Statistical analyses included experimental and clinico-pathological variables , as well as disease-free ( DFS ) and overall ( OS ) survival . RESULTS : P11836 -positive B lymphocytes were present in 57.7 % ( stromal ) /33.0 % ( intraepithelial ) and CD3-positive T lymphocytes in 99.0 % ( stromal ) /90.2 % ( intraepithelial ) of ovarian carcinomas . Intraepithelial CD3-positive T lymphocytes were correlated with improved DFS in optimally debulked patients ( P=0.0402 ) . Intraepithelial CD8-positive T lymphocytes were correlated with improved OS in all optimally debulked patients ( P=0.0201 ) and in those undergoing paclitaxel/carboplatin therapy ( P=0.0092 ) . Finally , rarified and clonal TCRgamma gene rearrangements were detected in 37 out of 93 ( 39.8 % ) and 15 out of 93 ( 16.1 % ) cases , respectively . This was marginally associated with improved DFS ( P=0.0873 ) . Despite a significant correlation of P04626 /neu status and intraepithelial CD8-positive lymphocytes ( P=0.0264 ) , this was non-directional ( R=-0.257 ; P=0.0626 ) . CONCLUSION : Improved survival of ovarian cancer patients is related to the infiltration , clonal selection and intraepithelial persistence of T lymphocytes . 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 . Oncotargets in different renal cancer subtypes . Renal cell cancer is a heterogeneous group of cancers with different histologic subtypes . The majority of renal tumors in adults are clear cell renal cell carcinomas , which are characterized by von Hippel- Lindau ( P40337 ) gene alterations . Recent advances in defining the genetic landscape of renal cancer has shown the genetic heterogeneity of clear cell renal cell carcinomas ( ccRCC ) and the presence of at least 3 additional ccRCC tumor suppressor genes on chromosome 3p . Due to inactivation of P40337 , renal cancer cells produce the HIF-responsive growth factor P15692 . The PI3K -- mTORC1 signaling axis also represents a target for therapy . The new systemic therapies , including tyrosine kinase inhibitors , monoclonal antibodies , and P42345 inhibitors , aim to suppress angiogenesis with vascular endothelial growth factor as a target . Various P15692 -inhibitors are approved for the treatment of ccRCC and we discuss recent advancements in the treatment of metastatic ccRCC . Other gene alterations have been identified in hereditary cancer syndromes , e.g. Q8NFG4 , Q92574 , P49815 , P19532 , P19484 , O75030 , FH , P21912 , O14521 , MET , and P60484 and we review their role in renal tumor carcinogenesis , prognosis , and targeted therapy . By reviewing the associations between morphologic features and molecular genetics of renal cancer we provide insight into the basis for targeted renal cancer therapy . Identification of candidate small-molecule therapeutics to cancer by gene-signature perturbation in connectivity mapping . Connectivity mapping is a recently developed technique for discovering the underlying connections between different biological states based on gene-expression similarities . The sscMap method has been shown to provide enhanced sensitivity in mapping meaningful connections leading to testable biological hypotheses and in identifying drug candidates with particular pharmacological and/or toxicological properties . Challenges remain , however , as to how to prioritise the large number of discovered connections in an unbiased manner such that the success rate of any following-up investigation can be maximised . We introduce a new concept , gene-signature perturbation , which aims to test whether an identified connection is stable enough against systematic minor changes ( perturbation ) to the gene-signature . We applied the perturbation method to three independent datasets obtained from the GEO database : acute myeloid leukemia ( AML ) , cervical cancer , and breast cancer treated with letrozole . We demonstrate that the perturbation approach helps to identify meaningful biological connections which suggest the most relevant candidate drugs . In the case of AML , we found that the prevalent compounds were retinoic acids and Q07869 activators . For cervical cancer , our results suggested that potential drugs are likely to involve the P00533 pathway ; and with the breast cancer dataset , we identified candidates that are involved in prostaglandin inhibition . Thus the gene-signature perturbation approach added real values to the whole connectivity mapping process , allowing for increased specificity in the identification of possible therapeutic candidates . Paullones are potent inhibitors of glycogen synthase kinase-3beta and cyclin-dependent kinase 5/p25 . Paullones constitute a new family of benzazepinones with promising antitumoral properties . They were recently described as potent , DB00171 -competitive , inhibitors of the cell cycle regulating cyclin-dependent kinases ( CDKs ) . We here report that paullones also act as very potent inhibitors of glycogen synthase kinase-3beta ( GSK-3beta ) ( IC50 : 4-80 nM ) and the neuronal Q00535 /p25 ( IC50 : 20-200 nM ) . These two enzymes are responsible for most of the hyperphosphorylation of the microtubule-binding protein tau , a feature observed in the brains of patients with Alzheimer 's disease and other neurodegenerative ' taupathies ' . DB04014 , the most active paullone , was demonstrated to act by competing with DB00171 for binding to GSK-3beta . DB04014 inhibits the phosphorylation of tau in vivo at sites which are typically phosphorylated by GSK-3beta in Alzheimer 's disease . DB04014 also inhibits the Q00535 /p25-dependent phosphorylation of Q9UD71 in mouse striatum slices in vitro . This dual specificity of paullones may turn these compounds into very useful tools for the study and possibly treatment of neurodegenerative and proliferative disorders . Genetic ablation of epidermal P00533 reveals the dynamic origin of adverse effects of anti- P00533 therapy . Cancer patients treated with anti- P00533 ( epidermal growth factor receptor ) drugs often develop a dose-limiting pruritic rash of unknown etiology . The aims of our study were to define causal associations from a clinical study of cutaneous and systemic changes in patients treated with gefitinib and use these to develop and characterize a mouse model that recapitulates the human skin rash syndrome caused by anti- P00533 therapy . We examined the patients ' plasma before and after treatment with gefitinib and documented changes in chemokines and leukocyte counts associated with the extent of rash or the presence of pruritus . We established a parallel mouse model by ablating P00533 in the epidermis . These mice developed skin lesions similar to the human rash . Before lesion development , we detected increased mRNA expression of chemokines in the skin associated with early infiltration of macrophages and mast cells and later infiltration of eosinophils , T cells , and neutrophils . As the skin phenotype evolved , changes in blood counts and circulating chemokines reproduced those seen in the gefitinib-treated patients . Crossing the mutant mice with mice deficient for tumor necrosis factor-α ( P01375 -α ) receptors , MyD88 , NOS2 , P41597 , T cells , or B cells failed to reverse the skin phenotype . However , local depletion of macrophages provided partial resolution , suggesting that this model can identify targets that may be effective in preventing the troublesome and dose-limiting skin response to anti- P00533 drugs . These results highlight the importance of P00533 signaling in maintaining skin immune homeostasis and identify a macrophage contribution to a serious adverse consequence of cancer chemotherapy . Q14703 lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 . DB03203 1-phosphate ( Q14703 ) and P21453 ( P21453 ) play an important role in the egress of mature P01730 or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY720 ) , an P21453 functional antagonist , induced significant accumulation of CD62L(high) Q07108 (low) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti- P21453 antibody revealed that P21453 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration . 2-Acetyl-4-tetrahydroxybutylimidazole ( THI ) , an Q14703 lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 ) . At 6h after THI administration , P21453 -expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla , suggesting Q14703 lyase expression in the cells constructing thymic medullary P15151 . To determine the cells expressing Q14703 lyase in the thymus , we newly established a mAb ( YK19-2 ) specific for mouse Q14703 lyase . Immunohistochemical staining with YK19-2 revealed that Q14703 lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 , Q14703 lyase was expressed in ER-TR7-positive cells ( reticular fibroblasts and pericytes ) and CD31-positive vascular endothelial cells . Our findings suggest that Q14703 lyase expressed in the thymic medullary P15151 keeps the tissue Q14703 concentration low around the vessels and promotes thymic egress via up-regulation of P21453 . Q9UEF7 gene deficiency causes salt-sensitive hypertension via monocyte chemotactic protein-1/CC chemokine receptor 2-mediated inflammation . Q9UEF7 ( KL ) is a newly discovered aging suppressor gene . In mice , the KL gene extends the lifespan when overexpressed and shortens the lifespan when disrupted . This study investigated if KL deficiency affects BP and salt sensitivity using KL mutant heterozygous ( +/- ) mice and wild-type ( WT ) mice ( 9 weeks of age , 16 mice per group ) . Notably , systolic BP in KL(+/-) mice began to increase at the age of 15 weeks , reached a peak level at the age of 17 weeks , and remained elevated thereafter , whereas systolic BP remained consistent in WT mice . High salt ( HS ) intake further increased BP in KL(+/-) mice but did not affect BP in WT mice . Blockade of CC chemokine receptor 2 ( P41597 ) , involved in monocyte chemotaxis , by a specific P41597 antagonist ( DB05130 ) abolished the HS-induced increase in BP in KL(+/-) mice . Furthermore , HS loading substantially increased the expression of monocyte chemotactic protein-1 and the infiltration of macrophages and T cells in kidneys in KL(+/-) mice , and treatment with DB05130 abolished these effects . Treatment of KL(+/-) mice with DB05130 also attenuated the increased renal expressions of serum glucocorticoid-regulated kinase 1 , thiazide-sensitive NaCl cotransporter , and DB00171 synthase β along with the renal structural damage and functional impairment induced by HS loading . In conclusion , KL deficiency caused salt-sensitive hypertension and renal damage by P41597 -mediated inflammation . Identification and characterization of mouse Erbb2 gene in silico . The Q9UD71 - Q14849 - O15273 -PNMT-MGC9753- P04626 -MGC14832- Q14451 locus on human chromosome 17q12 is frequently amplified in human gastric and breast cancer . We have recently identified and characterized human MGC9753 ( also known as wild-type CAB2 ) and mouse Mgc9753 . Here , we identified and characterized mouse Erbb2 gene by using bioinformatics . BLAST programs revealed that mouse AK031099 cDNA was derived from mouse Erbb2 gene . Because AK031099 cDNA showed 806 C --> A nucleotide substitution compared with mouse genome draft sequences and mouse Erbb2 ESTs , the nucleotide sequence of mouse Erbb2 cDNA was determined in silico by correcting 806 A of AK031099 cDNA to C . Nucleotide position 48-3818 of mouse Erbb2 cDNA was the coding region . Mouse Erbb2 gene , consisting of 27 exons , was located within the Ppp1r1b-Grb7 locus on the mouse chromosome 11 . Mouse Erbb2 protein ( 1256 aa ) showed 87.5 % total-amino-acid identity with human P04626 protein , and 95.2 % total-amino-acid identity with rat Erbb2 protein . Mouse Ppp1r1b-Grb7 locus and human Ppp1r1b-Grb7 locus were evolutionarily conserved in the order and the orientation of genes therein . Nucleotide and amino-acid substitution rates of Neurod2 located centromeric to the Ppp1r1b-Grb7 locus were significantly lower than others within the Ppp1r1b-Grb7 locus . This is the first report on the complete coding sequence of mouse Erbb2 gene as well as on the comprehensive comparison of Ppp1r1b-Grb7 locus within the human and mouse genomes . Expression of the RNA helicase O00571 and the hypoxia response in breast cancer . AIMS : O00571 is an RNA helicase that has antiapoptotic properties , and promotes proliferation and transformation . In addition , O00571 was shown to be a direct downstream target of Q9BYW2 α ( the master regulatory of the hypoxia response ) in breast cancer cell lines . However , the relation between O00571 and hypoxia has not been addressed in human tumors . In this paper , we studied the relation between O00571 and the hypoxic responsive proteins in human breast cancer . METHODS AND RESULTS : O00571 expression was investigated by immunohistochemistry in breast cancer in comparison with hypoxia related proteins Q9BYW2 α , P11166 , Q16790 , P00533 , P04626 , Akt1 , P98177 , p53 , ERα , Q8N668 , P16591 kinase , Q13526 , P12830 , P38936 , p27 , P02787 receptor , O43524 , c- DB00134 and Notch1 . O00571 was overexpressed in 127 of 366 breast cancer patients , and was correlated with overexpression of Q9BYW2 α and its downstream genes Q16790 and P11166 . Moreover , O00571 expression correlated with hypoxia-related proteins P00533 , P04626 , P98177 , ERα and c- DB00134 in a Q9BYW2 α dependent fashion , and with Q8N668 , P16591 kinase , Akt1 , P12830 , P02786 and O43524 independent of Q9BYW2 α . CONCLUSIONS : In invasive breast cancer , expression of O00571 was correlated with overexpression of Q9BYW2 α and many other hypoxia related proteins , pointing to a distinct role for O00571 under hypoxic conditions and supporting the oncogenic role of O00571 which could have clinical implication for current development of O00571 inhibitors . P42345 inhibitors and the anti-diabetic biguanide metformin : new insights into the molecular management of breast cancer resistance to the P04626 tyrosine kinase inhibitor lapatinib ( DB01259 ) . The small molecule P04626 tyrosine kinase inhibitor ( TKI ) lapatinib ( DB01259 ) is approved for the therapy of patients with P04626 -positive breast carcinomas who have progressed on trastuzumab ( Herceptin ) . Unfortunately , the efficacy of this P04626 TKI is limited by both primary ( inherent ) and acquired resistance , the latter typically occurring within 12 months of starting therapy . One of the key factors limiting our understanding of the mechanisms involved in lapatinib resistance is the lack of published preclinical models . We herein review lapatinib-refractory models recently developed at the bench and the survival pathways discovered . As hyperactivation of the pharmacologically targetable PI3K/ P42345 /p70S6K1 axis appears to be central to the occurrence of lapatinib resistance , preclinical data showing enhanced antitumour effects when combining lapatinib with P42345 inhibitors ( e.g. , rapamycin analogues and DB00238 -BEZ235 ) highlight the importance of translational work to yield clinically useful regimens capable of delaying or treating lapatinib resistance . The unexpected ability of the anti-type II diabetes drug metformin to inactivate P42345 and decrease p70S6K1 activity further reveals that this biguanide , generally considered non-toxic and remarkably inexpensive , might be considered for new combinatorial lapatinib-based protocols in P04626 -overexpressing breast cancer patients . 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 . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . P02787 enhances the antiproliferative effect of aluminum on osteoblast-like cells . DB01370 ( Al ) retention in the body can cause metabolic bone disease . This disorder is characterized by reductions in the number of osteoblasts , a feature that suggests a disturbance in bone cell proliferation or differentiation . Because Al as well as iron ( Fe ) can bind to transferrin ( TF ) in plasma , the role of TF as a modifier of osteoblast proliferation was examined in UMR-106-01 osteoblast-like cells by measuring the incorporation of tritiated thymidine ( [ 3H ] -TdR ) into DNA ( counts.min-1.microgram cell protein-1 , means +/- SE ) during 48-h incubations in serum-free medium ( SFM ) . In the absence of TF , DNA synthesis decreased when media levels of Al exceeded 6-10 microM . The mitogenic response to physiological levels of unsaturated TF ( apo-TF ) was attenuated however during incubations with TF that was partially saturated with Al ( Al-TF ) . A similar inhibitory response was seen in cells incubated with the antiproliferative agent gallium ( Ga ) when added to SFM as partially saturated Ga-TF . TF produced a shift to the left in the inhibitory dose-response curve to Al in osteoblast-like cells ; thus , DNA synthesis decreased at substantially lower media concentrations of Al in cells grown in SFM containing partially saturated Al-TF . The results indicate that TF is an important determinant of the inhibitory effect of Al on DNA synthesis by osteoblast-like cells at the micromolar levels of Al that can occur in plasma in vivo . DB00139 dehydrogenase ( SDHx ) mutations in pituitary tumors : could this be a new role for mitochondrial complex II and/or Krebs cycle defects ? DB00139 dehydrogenase ( SDH ) or mitochondrial complex II is a multimeric enzyme that is bound to the inner membrane of mitochondria and has a dual role as it serves both as a critical step of the tricarboxylic acid or Krebs cycle and as a member of the respiratory chain that transfers electrons directly to the ubiquinone pool . Mutations in SDH subunits have been implicated in the formation of familial paragangliomas ( PGLs ) and/or pheochromocytomas ( PHEOs ) and in Carney-Stratakis syndrome . More recently , SDH defects were associated with predisposition to a Cowden disease phenotype , renal , and thyroid cancer . We recently described a kindred with the coexistence of familial PGLs and an aggressive GH-secreting pituitary adenoma , harboring an O14521 mutation . The pituitary tumor showed loss of heterozygosity at the O14521 locus , indicating the possibility that O14521 's loss was causatively linked to the development of the neoplasm . In total , 29 cases of pituitary adenomas presenting in association with PHEOs and/or extra-adrenal PGLs have been reported in the literature since 1952 . Although a number of other genetic defects are possible in these cases , we speculate that the association of PHEOs and/or PGLs with pituitary tumors is a new syndromic association and a novel phenotype for SDH defects .

MH_train_1195

MH_train_1195

MH_train_1195

interacts_with DB00912?

multiple_choice

Association of novel SNPs in the candidate genes affecting caprine milk fatty acids related to human health . In the present investigation , 618 milk samples of Sirohi breed of goat were collected , and analyzed for conjugated linoleic acid ( DB01211 , C18:2 ) and other fatty acids . The DB01211 in studied goat milk samples was 4.87 mg/g of milk fat and C18:2 cis-9 , trans-11 contributes 2.9 mg/g of milk fat and trans10 cis12 contributes 0.82 mg/g of milk fat . The saturated fatty acids in the milk accounted for 69.55 % and unsaturated fatty acid accounted for 28.50 % . The unsaturated fatty acid was constituted by monounsaturated fatty acid ( 24.57 % ) and polyunsaturated fatty acids ( 3.96 % . ) . The major contribution ( 45.56 % ) in total fatty acid was of C12:0 , C14:0 and C16:0 . C18:0 and short chain ones ( C4:0 , P13671 :0 , Q99618 :0 , and Q99622 :0 ) have a neutral or cholesterol-decreasing effect . The DNA sequence analysis of the genes ( O75907 , SCAP , P37231 , OLR , P05413 and PRL ) in a random panel of 8 Sirohi goats revealed 38 SNPs across the targeted regions . Out of the studied SNPs ( 38 ) across these genes , 22 SNPs had significant effect on one or a group of fatty acids including DB01211 . The genotypes at these loci showed significant differences in the least square means of a particular fatty acid or a group of fatty acids including DB01211 and its isomers . Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 ( Q16739 ) is an approved drug target for the treatment of Gaucher disease and is considered as a valid target for combating other human pathologies , including type 2 diabetes . The clinical drug N-butyldeoxynojirimycin ( DB00419 ) is thought to inhibit through mimicry of its substrate , ceramide . In this work we demonstrate that , in contrast to what is proposed in this model , the P06681 -hydroxyl of the deoxynojirimycin core is important for Q16739 inhibition . Here we show that P13671 -OH appears of less important , which may set guidelines for the development of Q16739 inhibitors that have less affinity ( in comparison with DB00419 ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide . Second generation sequencing of the mesothelioma tumor genome . The current paradigm for elucidating the molecular etiology of cancers relies on the interrogation of small numbers of genes , which limits the scope of investigation . Emerging second-generation massively parallel DNA sequencing technologies have enabled more precise definition of the cancer genome on a global scale . We examined the genome of a human primary malignant pleural mesothelioma ( MPM ) tumor and matched normal tissue by using a combination of sequencing-by-synthesis and pyrosequencing methodologies to a 9.6X depth of coverage . Read density analysis uncovered significant aneuploidy and numerous rearrangements . Method-dependent informatics rules , which combined the results of different sequencing platforms , were developed to identify and validate candidate mutations of multiple types . Many more tumor-specific rearrangements than point mutations were uncovered at this depth of sequencing , resulting in novel , large-scale , inter- and intra-chromosomal deletions , inversions , and translocations . Nearly all candidate point mutations appeared to be previously unknown SNPs . Thirty tumor-specific fusions/translocations were independently validated with PCR and Sanger sequencing . Of these , 15 represented disrupted gene-encoding regions , including kinases , transcription factors , and growth factors . One large deletion in Q8N608 resulted in altered transcription and expression of Q8N608 transcripts in a set of 53 additional MPM tumors correlated with survival . Additionally , three point mutations were observed in the coding regions of Q9C056 , a transcription regulator , and Q6P4R8 , a DNA-binding protein involved in modulating P19838 . Several regions containing genes such as Q9H0N5 and P00374 , which are involved in growth factor signaling and nucleotide synthesis , respectively , were selectively amplified in the tumor . Second-generation sequencing uncovered all types of mutations in this MPM tumor , with DNA rearrangements representing the dominant type . Effects of P37231 knock-down and hyperglycemia on insulin signaling in vascular smooth muscle cells from hypertensive rats . Peroxisome proliferator-activated receptor ( Q07869 ) -gamma , a target in the treatment of diabetes , improves insulin sensitivity and exerts cardiovascular protective effects by mechanisms that are not completely elucidated . To investigate underlying molecular mechanisms responsible for P37231 -associated vascular insulin signaling in hypertension , we tested whether P37231 downregulation in vascular smooth muscle cells ( VSMC ) from WKY and SHRSP rats would decrease insulin signaling and glucose uptake and whether this response would be worsened by hyperglycemia to a greater extent in VSMC of hypertensive origin . Passaged mesenteric artery VSMC grown in euglycemic ( 5.5 mmol/L ) or hyperglycemic media ( 25.0 mmol/L ) were treated with P37231 -siRNA ( 5 nmol/L ) , P37231 antagonist ( GW-9662 , 10 micromol/L ) , or P37231 activator ( rosiglitazone , 10 micromol/L ) in the presence or absence of insulin ( 100 nmol/L ) . Immunoblotting revealed that hyperglycemia and P37231 inhibition significantly ( P < 0.001 ) decreased insulin-stimulated insulin receptor ( IR ) -beta , Akt , and glycogen synthase kinase ( GSK ) -3beta phosphorylation , whereas phosphotyrosine phosphatase ( PTP ) -1B expression was increased in VSMC from both strains . These effects were more pronounced in SHRSP under hyperglycemia . Rosiglitazone tended to increase insulin-mediated IR-beta , Akt , and GSK-3beta phosphorylation in VSMC from both strains , whereas insulin-induced P18031 expression was reduced by hyperglycemia . P01308 -stimulated P14672 expression and glucose transport were attenuated by hyperglycemia in both VSMC . These data suggest that P37231 inhibition results in decreased insulin signaling , particularly in SHR , in an IR-beta phosphorylation-dependent manner . Embryo number and periconceptional undernutrition in the sheep have differential effects on adrenal epigenotype , growth , and development . Exposure to poor maternal nutrition around the time of conception results in an early prepartum activation of the fetal pituitary-adrenal axis and in increased adrenal growth and stress response after birth associated with epigenetic changes in a differentially methylated region ( DMR ) of adrenal P01344 /H19 . We have determined the effects of maternal undernutrition during the periconceptional period ( PCUN : 70 % of control intake from 60 days before until 6 days after conception ) and early preimplantation period ( PIUN : 70 % of control intake for 6 days after conception ) on fetal plasma DB01285 and cortisol concentrations and fetal adrenal Q01718 , P49675 , 3βHSD , CYP11B , P05093 , TGFβ1 , IGF1 , P08069 , P01344 , and P11717 mRNA expression and the methylation level of sites within the DMRs of P01344 /H19 and P11717 in the adrenal of twin and singleton fetuses at 136-138 days gestation . Being a twin resulted in a delayed prepartum increase in fetal DB01285 and in a lower cortisol response to P06850 in the control but not PCUN and PIUN groups . PCUN , but not PIUN , resulted in an increase in adrenal weight and P05093 expression in singletons , a decrease in adrenal P01344 expression in singletons , and an increase in adrenal P11717 expression in both twins and singletons . P01344 /H19 and P11717 DMR methylation levels and Q01718 expression were lower in the twin adrenal . Thus , exposure of the oocyte and embryo to maternal undernutrition or to the environment of a twin pregnancy have differential effects on epigenetic and other factors that regulate fetal adrenal growth and P01344 and P11717 expression . [ Synthesis of ( 2'-bromo-4 ' , 5'-dimethoxy-phenyl ) -(2,3-dibromo-4,5-dimethoxy-phenyl)-methane as P18031 inhibitor ] . OBJECTIVE : To synthesize (2'-bromo-4',5'-dimethoxy-phenyl)- ( 2,3- dibromo-4,5-dimethoxy-phenyl ) -methane ( 6 ) as protein tyrosine phosphatase 1B ( P18031 ) inhibitor . METHOD : DB04088 was synthesized by Friedel-Crafts reaction , bromination and decarbonylation and screened inhibitory activity against P18031 by the colorimetric assay . The structure of synthetic intermediates and target product were identified on the basis of spectral analysis . RESULT : DB04088 was synthesized successfully in four steps and evaluated for its P18031 inhibitory activity , the screening result shown that compound 6 displayed good inhibitory activity against P18031 . CONCLUSION : The target compound 6 was synthesized with the overall yield of 20 % , which was a new compound and shown good inhibitory activity against P18031 ( inhibition 40.16 % at 5 mg x L(-1) ) . 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 . 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 . Inhibitors of the interaction between P04275 and platelet GPIb/IX/V . The formation of platelet-rich thrombi , a critical step in the pathogenesis of atherothrombotic events , is a multistep process involving several components , among which von Willebrand Factor ( P04275 ) plays a central role . Ruptured atherosclerotic plaques expose subendothelial matrix proteins which bind P04275 that represents a bridge between the injured blood vessel and activated platelets , playing a crucial role in platelet adhesion and aggregation , especially in conditions of high-shear rate . Due to these peculiarities , the binding of P04275 to GPIbα is an attractive drug target . Here we summarize the present knowledge on the different classes of drugs targeting the P04275 -GPIb interaction and we give an account of their level of clinical development . In particular , the following compounds are discussed : AJW200 , an IgG4 humanized monoclonal antibody against P04275 -A1 ; 82D6A3 , a monoclonal antibody against P04275 -A3 ; Q96JZ2 -0081 and Q96JZ2 -0681 , bivalent humanized nanobodies targeting the P04275 -A1 domain ; DB05202 and its advanced formulation ARC15105 , second-generation aptamers that bind the P04275 -A1 domain ; h6B4-Fab , a murine monoclonal antibody , and GPG-290 , a recombinant chimeric protein , both directed against GPIbα . 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 . DNA hypomethylation , transient neonatal diabetes , and prune belly sequence in one of two identical twins . One known genetic mechanism for transient neonatal diabetes is loss of methylation at 6q24 . The etiology of prune belly sequence is unknown but a genetic defect , affecting the mesoderm from which the triad abdominal muscle hypoplasia , urinary tract abnormalities , and cryptorchidism develop , has been suggested . We investigated a family , including one twin , with transient neonatal diabetes and prune belly sequence . Autoantibody tests excluded type 1 diabetes . Microsatellite marker analysis confirmed the twins being monozygotic . We identified no mutations in Q9NU63 , Q14654 , Q09428 , GCK , P20823 , P35680 , Q9Y261 , P52945 , O43316 , or O60481 . The proband had loss of methylation at the 6q24 locus TNDM and also at the loci P11717 , O95661 , and Q5EB52 , while the other family members , including the healthy monozygotic twin , had normal findings . The loss of methylation on chromosome 6q24 and elsewhere may indicate a generalized maternal hypomethylation syndrome , which accounts for both transient neonatal diabetes and prune belly sequence . 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 . Prostanoid production in Saccharomyces cerevisiae provides a novel assay for nonsteroidal anti-inflammatory drugs . Prostanoids are a large family of lipid mediators originating from prostaglandin H synthase ( PGHS ) activity on the 20-carbon polyunsaturated fatty acids dihomo-gamma-linolenic acid ( DB00154 ) , arachidonic acid ( AA ) and eicosapentaenoic acid . The two mouse PGHS isoforms , P23219 and P35354 , were expressed in Saccharomyces cerevisiae ( yeast ) , as was a signal-peptide-deleted version of P23219 ( PGHS-1MA ) . P23219 showed high activity with both AA and DB00154 as substrate , whereas P35354 activity was high with DB00154 but low with AA . Signal peptide removal reduced the activity of PGHS-1MA by > 50 % relative to P23219 , but the residual activity indicated that correct targeting to the lumen of the endoplasmic reticulum may not be necessary for enzyme function . Coexpression of P23219 with cDNAs encoding mouse prostaglandin I synthase and thromboxane A synthase , and with Trypanosoma brucei genomic DNA encoding prostaglandin F synthase in AA-supplemented yeast cultures resulted in production of the corresponding prostanoids , prostaglandin I(2) , thromboxane A(2) and prostaglandin F(2alpha) . The inhibitory effects of nonsteroidal anti-inflammatory drugs ( NSAIDs ) on prostanoid production were tested on yeast cells expressing P23219 in AA-supplemented culture . Dose-dependent inhibition of prostaglandin H(2) production by aspirin , ibuprofen and indomethacin demonstrated the potential utility of this simple expression system in screening for novel NSAIDs . 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 . Effects on thrombin generation of single injections of DB02351 in patients with calf vein thrombosis . STUDY OBJECTIVE : To determine whether single injections of DB02351 , a direct thrombin inhibitor , can inhibit thrombin generation in patients with calf vein thrombosis and , if so , if the inhibition is sustained . DESIGN : Phase II open label cohort study . SETTING : Tertiary-care referral centres , university affiliated hospitals . PATIENTS : 10 patients with venographically-demonstrated calf vein thrombosis . INTERVENTION : Patients received a single injection of DB02351 , either 1.0 mg/kg subcutaneously or 0.6 mg/kg as a 15 min intravenous infusion . P00734 fragment ( F1++2 ) levels , as an index of thrombin generation , were measured before as well as 6 h post- and 24 h post- DB02351 administration . Patients were followed with non-invasive tests to detect thrombus extension into the proximal veins . RESULTS : There was a significant reduction in the levels of F1+2 with both regimens , 6 h after DB02351 . The F1+2 levels 24 h post- DB02351 showed a significant increase relative to the 6 h post- DB02351 results . One patient developed thrombus extension into the popliteal vein and was treated with conventional anticoagulants . CONCLUSION : The single injections of DB02351 used in the study produced incomplete and temporary suppression of F1+2 . Complete and permanent inhibition of thrombin generation with DB02351 in patients with calf vein thrombosis may require higher doses , multiple subcutaneous injections and/or prolonged intravenous infusion . A mechanism for the synergistic antimalarial action of atovaquone and proguanil . A combination of atovaquone and proguanil has been found to be quite effective in treating malaria , with little evidence of the emergence of resistance when atovaquone was used as a single agent . We have examined possible mechanisms for the synergy between these two drugs . While proguanil by itself had no effect on electron transport or mitochondrial membrane potential ( DeltaPsim ) , it significantly enhanced the ability of atovaquone to collapse DeltaPsim when used in combination . This enhancement was observed at pharmacologically achievable doses . DB01131 acted as a biguanide rather than as its metabolite cycloguanil ( a parasite dihydrofolate reductase [ P00374 ] inhibitor ) to enhance the atovaquone effect ; another P00374 inhibitor , pyrimethamine , also had no enhancing effect . DB01131 -mediated enhancement was specific for atovaquone , since the effects of other mitochondrial electron transport inhibitors , such as myxothiazole and antimycin , were not altered by inclusion of proguanil . Surprisingly , proguanil did not enhance the ability of atovaquone to inhibit mitochondrial electron transport in malaria parasites . These results suggest that proguanil in its prodrug form acts in synergy with atovaquone by lowering the effective concentration at which atovaquone collapses DeltaPsim in malaria parasites . This could explain the paradoxical success of the atovaquone-proguanil combination even in regions where proguanil alone is ineffective due to resistance . The results also suggest that the atovaquone-proguanil combination may act as a site-specific uncoupler of parasite mitochondria in a selective manner . Pharmacology of recombinant low-voltage activated calcium channels . Several types of voltage- or ligand-activated calcium channels contribute to the excitability of neuronal cells . Low-voltage-activated ( LVA ) , T-type calcium channels are characterised by relatively negative threshold of activation and therefore they can generate low-threshold spikes , which are essential for burst firing . At least three different proteins form T-type calcium current in neurons : Ca(v)3.1 , Ca(v)3.2 and Q9P0X4 . Expression of these proteins in various brain regions is complementary . Individual channel types could be distinguished by different sensitivity towards inorganic cations . This inhibition can contribute to the toxicity of some heavy metals . Selective inhibition of T-type calcium channels by organic blockers may have clinical importance in some forms of epilepsy . DB01388 inhibits the expressed Ca(v2)3.1 , Ca(v)3.2 and Q9P0X4 channels in nanomolar concentrations with Q9P0X4 channel having lowest affinity . The sensitivity of the expressed Ca(v)3.1 channel to the antiepileptic drugs , valproate and ethosuximide , is low . Ca(v)3.1 channel is moderately sensitive to phenytoin . The Ca(v)3.2 channel is sensitive to ethosuximide , amlodipine and amiloride . All three LVA calcium channels are moderately sensitive to active metabolites of methosuximide , i.e. alpha-methyl-alpha-phenylsuccinimide . Several neuroleptics inhibit all three LVA channels in clinically relevant concentrations . All three channels are also inhibited by the endogenous cannabinoid anandamide . A high affinity peptide blocker for these Ca channels is the scorpion toxin kurtoxin which inhibits the Ca(v)3.1 and Ca(v)3.2 , but not the Q9P0X4 channel in nanomolar concentrations . DB06690 selectively inhibits the Ca(v)3.2 , but not the Ca(v)3.1 channel . The Ca(v)3.2 , but not the Ca(v)3.1 channel is potentiated by stimulation of Ca(2+)/ P62158 -dependent protein kinase . Electrostatic steering at acetylcholine binding sites . The electrostatic environments near the acetylcholine binding sites on the nicotinic acetylcholine receptor ( nAChR ) and acetylcholinesterase were measured by diffusion-enhanced fluorescence energy transfer ( DEFET ) to determine the influence of long-range electrostatic interactions on ligand binding kinetics and net binding energy . Changes in DEFET from variously charged Tb3+ -chelates revealed net potentials of -20 mV at the nAChR agonist sites and -14 mV at the entrance to the P22303 active site , in physiological ionic strength conditions . The potential at the alphadelta-binding site of the nAChR was determined independently in the presence of DB01199 to be -14 mV ; the calculated potential at the alphagamma-site was approximately threefold stronger than at the alphadelta-site . By determining the local potential in increasing ionic strength , Debye-Hückel theory predicted that the potentials near the nAChR agonist binding sites are constituted by one to three charges in close proximity to the binding site . Examination of the binding kinetics of the fluorescent acetylcholine analog dansyl- P13671 -choline at ionic strengths from 12.5 to 400 mM revealed a twofold decrease in association rate . Debye-Hückel analysis of the kinetics revealed a similar charge distribution as seen by changes in the potentials . To determine whether the experimentally determined potentials are reflected by continuum electrostatics calculations , solutions to the nonlinear Poisson-Boltzmann equation were used to compute the potentials expected from DEFET measurements from high-resolution models of the nAChR and P22303 . These calculations are in good agreement with the DEFET measurements for P22303 and for the alphagamma-site of the nAChR . We conclude that long-range electrostatic interactions contribute -0.3 and -1 kcal/mol to the binding energy at the nAChR alphadelta- and alphagamma-sites due to an increase in association rates . 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 . Suppressed Ca2+/ P62158 /CaMKII-dependent K( DB00171 ) channel activity in primary afferent neurons mediates hyperalgesia after axotomy . Painful axotomy decreases K( DB00171 ) channel current ( IK( DB00171 ) ) in primary afferent neurons . Because cytosolic Ca(2+) signaling is depressed in injured dorsal root ganglia ( Q86YR7 ) neurons , we investigated whether Ca(2+)-calmodulin ( P62158 ) -Ca(2+)/ P62158 -dependent kinase II ( CaMKII ) regulates IK( DB00171 ) in large Q86YR7 neurons . Immunohistochemistry identified the presence of K( DB00171 ) channel subunits Q09428 , SUR2 , and Kir6.2 but not Kir6.1 , and pCaMKII in neurofilament 200-positive Q86YR7 somata . Single-channel recordings from cell-attached patches revealed that basal and evoked IK( DB00171 ) by ionomycin , a Ca(2+) ionophore , is activated by CaMKII . In axotomized neurons from rats made hyperalgesic by spinal nerve ligation ( Q16658 ) , basal K( DB00171 ) channel activity was decreased , and sensitivity to ionomycin was abolished . Basal and Ca(2+)-evoked K( DB00171 ) channel activity correlated inversely with the degree of hyperalgesia induced by Q16658 in the rats from which the neurons were isolated . Inhibition of IK( DB00171 ) by glybenclamide , a selective K( DB00171 ) channel inhibitor , depolarized resting membrane potential ( O94763 ) recorded in perforated whole-cell patches and enhanced neurotransmitter release measured by amperometry . The selective K( DB00171 ) channel opener diazoxide hyperpolarized the O94763 and attenuated neurotransmitter release . Axotomized neurons from rats made hyperalgesic by Q16658 lost sensitivity to the myristoylated form of autocamtide-2-related inhibitory peptide ( AIPm ) , a pseudosubstrate blocker of CaMKII , whereas axotomized neurons from Q16658 animals that failed to develop hyperalgesia showed normal IK( DB00171 ) inhibition by AIPm . AIPm also depolarized O94763 in control neurons via K( DB00171 ) channel inhibition . Unitary current conductance and sensitivity of K( DB00171 ) channels to cytosolic DB00171 and ligands were preserved even after painful nerve injury , thus providing opportunities for selective therapeutic targeting against neuropathic pain . 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 .

MH_train_1196

MH_train_1196

MH_train_1196

interacts_with DB01030?

multiple_choice

Lipid-lowering properties of DB05317 , a squalene synthase inhibitor , in vivo and in vitro . 1. P37268 is the enzyme that converts farnesyl pyrophosphate to squalene in the cholesterol biosynthesis pathway . We examined the lipid-lowering properties of 1- [ [ ( 3R,5S ) -1-(3-acetoxy-2,2-dimethylpropyl)-7-chloro-5-(2,3-dimethoxyphenyl)-2-oxo-1,2,3,5-tetrahydro-4,1-benzoxazepin-3-yl ] acetyl ] piperidine-4-acetic acid ( DB05317 ) , a novel squalene synthase inhibitor . 2 . DB05317 inhibited hepatic cholesterol biosynthesis in rats ( ED(50) , 2.9 mg kg(-1) ) and showed lipid-lowering effects in beagle dogs , marmosets , cynomolgus monkeys and Wistar fatty rats . 3 . In marmosets , DB05317 ( 30 , 100 mg kg(-1) , p.o. , for 4 days ) lowered both plasma non-high-density lipoprotein ( HDL ) cholesterol and triglyceride , but did not affect plasma HDL cholesterol . On the other hand , atorvastatin ( 10 , 30 mg kg(-1) , p.o. , for 4 days ) lowered the levels of all these lipids . A correlation between decrease in triglyceride and increase in HDL cholesterol was observed , and DB05317 increased HDL cholesterol with a smaller decrease in triglyceride than did atorvastatin . 4 . DB05317 ( 60 mg kg(-1) , p.o. , for 15 days ) suppressed the rate of triglyceride secretion from the liver in hypertriglyceridemic Wistar fatty rats , which show an enhanced triglyceride secretion rate from the liver compared with their lean littermates . 5 . In HepG2 cells , DB05317 and its pharmacologically active metabolite , T-91485 , increased the binding of (125)I-low-density lipoprotein ( LDL ) to LDL receptors . 6 . These results suggest that DB05317 has clear hypolipidemic effects in animals via inhibition of hepatic triglyceride secretion and upregulation of LDL receptors , and that DB05317 might increase HDL cholesterol by decreasing triglyceride . Thus , DB05317 is expected to be useful for the treatment of dyslipidemia . DB06643 for bone diseases : translating bone biology into targeted therapy . Signalling of receptor activator of nuclear factor-κB ( Q9Y6Q6 ) ligand ( O14788 ) through Q9Y6Q6 is a critical pathway to regulate the differentiation and activity of osteoclasts and , hence , a master regulator of bone resorption . Increased O14788 activity has been demonstrated in diseases characterised by excessive bone loss such as osteoporosis , rheumatoid arthritis and osteolytic bone metastases . The development and approval of denosumab , a fully MAB against O14788 , has heralded a new era in the treatment of bone diseases by providing a potent , targeted and reversible inhibitor of bone resorption . This article summarises the molecular and cellular biology of the O14788 / Q9Y6Q6 system and critically reviews preclinical and clinical studies that have established denosumab as a promising novel therapy for metabolic and malignant bone diseases . We will discuss the potential indications for denosumab along with a critical review of safety and analyse its potential within the concert of established therapies . P11387 is a cofactor for c-Jun in the regulation of epidermal growth factor receptor expression and cancer cell proliferation . P11387 ( Topo I ) is a molecular target for the anticancer agent topotecan in the treatment of small cell lung cancer and ovarian carcinomas . However , the molecular mechanisms by which topotecan treatment inhibits cancer cell proliferation are unclear . We describe here the identification of Topo I as a novel endogenous interaction partner for transcription factor c-Jun . Reciprocal coimmunoprecipitation analysis showed that Topo I and c-Jun interact in transformed human cells in a manner that is dependent on JNK activity . c-Jun target gene epidermal growth factor receptor ( P00533 ) was identified as a novel gene whose expression was specifically inhibited by topotecan . Moreover , Topo I overexpression supported c-Jun-mediated reporter gene activation and both genetic and chemical inhibition of c-Jun converted cells resistant to topotecan-elicited P00533 downregulation . DB01030 -elicited suppression of proliferation was rescued by exogenously expressed P00533 . Furthermore , we demonstrate the cooperation of the JNK-c-Jun pathway , Topo I , and P00533 in the positive regulation of O75794 cell proliferation . Together , these results have identified transcriptional coactivator Topo I as a first endogenous cofactor for c-Jun in the regulation of cell proliferation . In addition , the results of the present study strongly suggest that inhibition of P00533 expression is a novel mechanism by which topotecan inhibits cell proliferation in cancer therapy . An overview of phenserine tartrate , a novel acetylcholinesterase inhibitor for the treatment of Alzheimer 's disease . Existing cholinesterase ( ChE ) inhibitor therapies for Alzheimer 's disease ( AD ) , while effective in improving cognitive , behavioral and functional impairments , do not alter disease progression . Novel drug design studies have focused on the classical ChE inhibitor , (-)-physostigmine , producing alterations in chemical composition and three-dimensional structure , which may offer an improved therapeutic index . The phenylcarbamate derivative , DB04892 , is a selective , non-competitive inhibitor of acetylcholinesterase ( P22303 ) . In vivo , DB04892 produces rapid , potent , and long-lasting P22303 inhibition . As a possible result of its preferential brain selectivity , DB04892 is significantly less toxic than (-)-physostigmine . In studies using the Stone maze paradigm , DB04892 has been shown to improve cognitive performance in both young learning-impaired and elderly rats . In addition to reducing inactivation of acetylcholine in the brain , DB04892 appears to have a second mode of action . Reduced secretion of beta-amyloid ( Abeta ) has been observed in cell lines exposed to DB04892 , occurring through translational regulation of beta-amyloid precursor protein ( beta- P05067 ) mRNA via a non-cholinergic mechanism . These in vitro findings appear to translate in vivo into animal models and humans . In a small study of patients with AD , DB04892 treatment tended to reduce beta- P05067 and Abeta levels in plasma samples . Clinical studies also reveal that DB04892 ( 5-10 mg b.i.d. ) had a favorable safety and pharmacological profile , produced significant improvements in cognitive function and was well tolerated in patients with AD treated for 12 weeks . Further randomized , double-blind , placebo-controlled Phase III studies assessing the efficacy , safety/tolerability and potential disease-modifying effects of DB04892 in patients with AD are currently ongoing . Differential binding of poly(ADP- DB01936 ) polymerase-1 and JunD/Fra2 accounts for O14788 -induced Tcirg1 gene expression during osteoclastogenesis . We studied Tcirg1 gene expression on O14788 -induced osteoclastic differentiation of the mouse model RAW264.7 cells . We identified a mechanism involving P09874 inhibition release and JunD/Fra-2 binding , which is responsible for Tcirg1 gene upregulation . INTRODUCTION : The Tcirg1 gene encodes the a3 isoform of the V-ATPase a subunit , which plays a critical role in the resorption activity of the osteoclast . Using serial deletion constructs of the Tcirg1 gene promoter , we performed a transcriptional study to identify factor(s) involved in the regulation of the O14788 -induced gene expression . MATERIALS AND METHODS : The promoter activity of serial-deletion fragments of the Tcirg1 gene promoter was monitored throughout the RAW264.7 cells differentiation process . We next performed sequence analysis , EMSA , UV cross-linking , qPCR , and gel supershift experiments to identify the factor(s) interacting with the promoter . RESULTS : A deletion of the -1297-1244 region led to the disappearance of the O14788 -induced promoter activity . EMSA experiments showed the binding of two factors that undergo differential binding on O14788 treatment . Supershift experiments led us to identify the dimer JunD/Fra-2 as the binding activity associated with the -1297/-1268 Tcirg1 gene promoter sequence in response to O14788 . Moreover , we observed poly(ADP-ribose) polymerase-1 ( P09874 ) binding to an adjacent site ( -1270/-1256 ) , and this interaction was disrupted after O14788 treatment . CONCLUSIONS : We provide data that identify junD proto-oncogene ( JunD ) and P15408 ( Fra-2 ) as the activator protein-1 ( AP-1 ) factors responsible for the O14788 -induced upregulation of the mouse Tcirg1 gene expression . Moreover , we identified another binding site for P09874 that might account for the repression of Tcirg1 gene expression in pre-osteoclastic cells . Design and synthesis of an orally bioavailable and selective peptide epoxyketone proteasome inhibitor ( PR-047 ) . Proteasome inhibition has been validated as a therapeutic modality in the treatment of multiple myeloma and non-Hodgkin 's lymphoma . DB08889 , an epoxyketone currently undergoing clinical trials in malignant diseases , is a highly selective inhibitor of the chymotrypsin-like ( CT-L ) activity of the proteasome . A chemistry effort was initiated to discover orally bioavailable analogues of carfilzomib , which would have potential for improved dosing flexibility and patient convenience over intravenously administered agents . The lead compound , 2-Me-5-thiazole- DB00133 (OMe)- DB00133 (OMe)- DB00120 -ketoepoxide ( 58 ) ( PR-047 ) , selectively inhibited CT-L activity of both the constitutive proteasome ( beta5 ) and immunoproteasome ( P28062 ) and demonstrated an absolute bioavailability of up to 39 % in rodents and dogs . It was well tolerated with repeated oral administration at doses resulting in > 80 % proteasome inhibition in most tissues and elicited an antitumor response equivalent to intravenously administered carfilzomib in multiple human tumor xenograft and mouse syngeneic models . The favorable pharmacologic profile supports its further development for the treatment of malignant diseases . Array comparative genomic hybridization , expression array , and protein analysis of critical regions on chromosome arms 1q , 7q , and 8p in adenocarcinomas of the gastroesophageal junction . Survival rates of adenocarcinomas of the gastroesophageal junction ( GEJ ) are low , because these tumors are generally in an advanced stage by the time they are detected . Chromosomal regions 1q32 , 7q21 , and 8p22 display critical alterations in GEJ cancers ; however , the genes underlying alterations in these genomic areas are largely unknown . To delineate overexpressed genes , we performed array comparative genomic hybridization ( aCGH ) and mRNA expression analysis of 15 GEJ adenocarcinoma samples using a fine-tiling cDNA array covering chromosome segments 1q31.3~q41 ( 193.9-215.8 Mb : 21.9 Mb ) , 7q11.23~q22.1 ( 72.3-103.0 Mb : 30.7 Mb ) , and 8p23.1~ P38936 .3 ( 11.1-20.7 Mb : 9.6 Mb ) . Based on a mRNA overexpression criterion , 11 genes were selected : P78545 and Q96JT2 on 1q ; P56749 , Q00534 , Q9HCE7 , O15143 , P17029 , P33993 , and Q7L5N1 on 7q ; and P37268 and P07858 on 8p . The protein expression levels were subsequently determined by immunohistochemical analysis of the cancer samples . There was a significant correlation between genomic amplification , mRNA , and protein expression or overexpression for Q00534 , a cell cycle regulator on 7q21.2 ( 92.1 Mb ; P < 0.01 ) ; other genes showed less stringent associations . In conclusion , using a straightforward approach we constructed a targeted gene profile for GEJ adenocarcinomas . Effective panitumumab treatment in patients with heavily pre-treated metastatic colorectal cancer : a case series . In heavily pre-treated patients with metastatic colorectal cancer ( mCRC ) , further chemotherapy has not demonstrated efficacy . DB01269 is indicated as monotherapy treatment of epidermal growth factor receptor ( P00533 ) -expressing , Kirsten ( K ) - DB01367 wild-type metastatic colorectal cancer after failure of fluoropyrimidine- , oxaliplatin- and irinotecan-containing regimens . However , panitumumab has not been specifically evaluated in patients following failure of a bevacizumab-containing regimen . One female and two male patients with mCRC presented with tumour recurrence in the para-aortic lymph nodes , the liver and the local presacral lymph nodes , respectively . The patients were confirmed to have K- DB01367 wild-type-expressing tumours . Following the failure of bevacizumab-containing chemotherapy regimens , all three patients received panitumumab monotherapy . DB01269 was well-tolerated . All the patients responded to panitumumab monotherapy in this late-stage setting . This patient series suggests that panitumumab can improve patient outcomes and may be an alternative treatment option in patients with mCRC who have received prior treatment with bevacizumab plus chemotherapy . DNA synthesis and neuronal apoptosis caused by familial Alzheimer disease mutants of the amyloid precursor protein are mediated by the P38936 activated kinase O75914 . Apoptotic pathways and DNA synthesis are activated in neurons in the brains of individuals with Alzheimer disease ( AD ) . However , the signaling mechanisms that mediate these events have not been defined . We show that expression of familial AD ( DB03147 ) mutants of the amyloid precursor protein ( P05067 ) in primary neurons in culture causes apoptosis and DNA synthesis . Both the apoptosis and the DNA synthesis are mediated by the P38936 activated kinase O75914 , a serine-threonine kinase that interacts with P05067 . A dominant-negative kinase mutant of O75914 inhibits the neuronal apoptosis and DNA synthesis ; this effect is abolished by deletion of the O75914 P05067 -binding domain or by coexpression of a peptide representing this binding domain . The involvement of O75914 specifically in DB03147 P05067 -mediated apoptosis rather than in general apoptotic pathways is suggested by the facts that a dominant-positive mutant of O75914 does not alone cause neuronal apoptosis and that the dominant-negative mutant of O75914 does not inhibit chemically induced apoptosis . Pertussis toxin , which inactivates the heterotrimeric G-proteins Go and Gi , inhibits the apoptosis and DNA synthesis caused by DB03147 P05067 mutants ; the apoptosis and DNA synthesis are rescued by coexpression of a pertussis toxin-insensitive Go . DB03147 P05067 -mediated DNA synthesis precedes DB03147 P05067 -mediated apoptosis in neurons , and inhibition of neuronal entry into the cell cycle inhibits the apoptosis . These data suggest that a normal signaling pathway mediated by the interaction of P05067 , O75914 , and Go is constitutively activated in neurons by DB03147 mutations in P05067 and that this activation causes cell cycle entry and consequent apoptosis . Association of DNA repair gene polymorphisms with response to platinum-based doublet chemotherapy in patients with non-small-cell lung cancer . PURPOSE : To identify polymorphisms in DNA repair genes that affect responses to platinum-based doublet chemotherapy in patients with non-small-cell lung cancer ( NSCLC ) . PATIENTS AND METHODS : In total , 640 patients with NSCLC who received platinum-based doublet chemotherapy in the National Cancer Center Hospital in Japan from 2000 to 2008 and whose responses were evaluated by Response Evaluation Criteria in Solid Tumors ( RECIST ) participated in a study of the association between response and genotypes for 30 single nucleotide polymorphisms ( SNPs ) in 27 DNA repair genes . Candidate SNPs were selected in a discovery set of 201 patients , and their associations were validated in an independent set of 439 patients by prespecified P value criteria . RESULTS : Homozygotes for the minor allele P04637 -72Pro of the Arg72Pro SNP in the P04637 gene showed a better response rate ( 54.3 % ) than those for the major allele P04637 -72Arg ( 29.1 % ; P = 4.4 × 10(-5) ) irrespective of therapeutic regimens , and minor allele homozygotes had significantly longer progression-free and overall survivals than major allele homozygotes ( hazard ratio [ HR ] , 0.85 ; 95 % CI , 0.74 to 0.98 ; P = .020 ; and HR , 0.86 ; 95 % CI , 0.74 to 0.99 ; P = .039 ) . Minor allele carriers for SNP Lys940Arg in the poly ( ADP-ribose ) polymerase 1 ( P09874 ) gene showed a better response rate to the paclitaxel regimen ( 45.8 % ) than to the gemcitabine regimen ( 10.5 % ; P for interaction = .019 ) . CONCLUSION : Polymorphisms in the P04637 and P09874 genes are involved in inter-individual differences in the response to platinum-based doublet chemotherapy in patients with NSCLC . Glucagon-like peptide 1 protects microvascular endothelial cells by inactivating the P09874 / P35228 /NO pathway . Increasing studies suggest that the activity of P0C6A0 might be of significant importance in the development of type 2 diabetes beyond its serum glucose-lowering effects . However , to date , the anti-apoptosis mechanism by which P0C6A0 acts on MILE SVEN 1 ( MS-1 ) cells has not been fully explored with regard to the intracellular signaling pathway . Increasing evidence shows that apoptosis of islet microvascular endothelial cells ( IMECs ) participates in the pathogenesis of diabetes . We wondered whether P0C6A0 exerts its anti-apoptosis effects by inactivating the P09874 / P35228 /NO pathway in oxidized low-density-lipoprotein ( oxLDL ) -induced apoptosis in mouse IMECs ( MS-1 cells ) , which may linked to P43220 / DB02527 levels . MTT assay revealed that 2-h pre-incubation with P0C6A0 markedly restored the oxLDL-induced loss of MS-1 viability in a concentration-dependent manner . This effect was accompanied by a significant decrease in intracellular nitric oxide ( NO ) activity . Moreover , P0C6A0 suppressed lipid peroxidation , restored the activities of endogenous antioxidants , and decreased the level of NO . Pre-incubating MS-1 cells with P0C6A0 reduced cell apoptosis . Finally , P0C6A0 could efficiently prevent the upregulation of poly(ADP-ribose) polymerase-1/nitrotyrosine and inducible NO synthase protein . Simultaneously , the expression of P43220 and the level of DB02527 was consistent with the administration of P0C6A0 . Our findings suggest that P0C6A0 can effectively protect MS-1 cells against oxLDL-induced apoptosis , which may be important in preventing the pathogenesis of diabetes mellitus . Regulation of fibrillin-1 gene expression by Sp1 . Mutations in the fibrillin-1 gene ( P35555 ) cause Marfan Syndrome ( MFS ) , a hereditary disorder of connective tissue . The transcription of P35555 has been reported to be driven by a short ultraconserved region ( SUPR ) in the 5' untranslated exon A of P35555 , but the nature of other factors involved in P35555 gene regulation has not been clarified . In this study , we characterized the transcription factors involved in P35555 gene regulation . The results show that Sp1 protein binds to two putative binding sites in the promoter of P35555 . Overexpression of Sp1 resulted in a significant increase in both promoter activity and P35555 mRNA level in P29320 293 cells , whereas inhibition or knockdown of Sp1 decreased P35555 gene expression . In addition , we found that Poly [ ADP-ribose ] polymerase 1 ( P09874 ) binds to the palindromic sequence TCTCGCGAGA in the ultraconserved region of the P35555 promoter and that the regulation of P35555 expression by P09874 is dependent on Sp1 . These results indicate that both Sp1 and P09874 contribute to P35555 gene expression . These observations add to our understanding of the transcriptional regulation of P35555 gene expression . Radiosensitization by inhibition of IkappaB-alpha phosphorylation in human glioma cells . To assess the role of nuclear factor kappaB ( NFKB ) in cellular radiosensitivity , three different IkappaB-alpha ( also known as P25963 ) expression plasmids , i.e. , S-IkappaB ( mutations at ( 32 , 36 ) DB00133 ) , Y-IkappaB ( a mutation at (42) DB00135 ) , and SY-IkappaB , were constructed and introduced into human brain tumor M054 cells . The clones were named as M054-S8 , M054- P28062 and M054-SY4 , respectively . Compared to the parental cell line , M054-S8 and M054- P28062 cells were more sensitive to X rays while M054-SY4 cells exhibited the greatest sensitivity . After treatment with N-acetyl- DB00149 - DB00149 -norleucinal , a proteasome inhibitor , the X-ray sensitivity of M054-S8 and M054-SY4 cells did not change , while that of M054- P28062 cells and the parental cells was enhanced . An increase in X-ray sensitivity accompanied by a decrease in translocation of NFKB to the nucleus in parental cells was observed after treatment with pervanadate , an inhibitor of tyrosine phosphatase , as well as in M054-S8 and M054-SY4 cells . Repair of potentially lethal damage ( PLD ) was observed in the parental cells but not in the clones . Four hours after irradiation ( 8 Gy ) , the expression of P04637 and phospho-p53 ( (15) DB00133 ) was induced in the parental cells but not in M054-S8 , M054- P28062 or M054-SY4 cells . Our data suggest that inhibition of IkappaB-alpha phosphorylation at serine or tyrosine acts independently in sensitizing cells to X rays . NFKB may play a role in determining radiosensitivity and PLD repair in malignant glioma cells ; P04637 may also be involved . DNA damage-related gene expression as biomarkers to assess cellular response after gamma irradiation of a human lymphoblastoid cell line . Since defects in molecular mechanisms controlling DNA repair , cell cycle checkpoint and apoptosis could modify cellular sensitivity to DNA damaging agents , we have conducted a multiparametric molecular analysis for better understanding the regulation pathways leading to cell survival or cell death after irradiation . Using a human lymphoblastoid cell line , we have analysed , following gamma irradiation ( 0.5 , 1 , 2 , 4 , 8 , 16 and 32 Gy , at 0.5 , 24 , 48 and 72 h after treatment ) , the correlation between proliferation , cell cycle analysis , apoptosis and micronuclei frequency with the expression of P04637 , P38936 , DNA LIGASE 1 , P12004 , Q07812 , O43927 -2 , Q16611 , P61803 , P42575 -Long and -Short forms mRNAs . We have found that whereas P04637 , Q16611 , P42575 -Short form , and P61803 were expressed at constant levels , P38936 , P12004 , Q07812 were up-regulated , P42575 -Long form , DNA LIGASE 1 , and BCL-2 were down-regulated . These modifications of expression were significantly correlated with doses , survival , proliferation , cell cycle delays , and apoptosis . A positive correlation of P38936 and Q07812 , and a borderline negative correlation with BCL-2 expressions were observed with micronuclei frequency for doses ranging from 0.5 to 4 Gy . In conclusion , our data clearly demonstrate that gene expression profiling , which is easier and more rapid to conduct than the assessments of classical phenotypic responses , could be useful to improve knowledge concerning pathways involved in cellular response to irradiation , knowing that such biomarkers could constitute tools to assess radio-sensitivity/radio-resistance . Oncogene ( 2000 ) 19 , 916 - 923 . Role of nitric oxide ( NO ) in pulmonary dysfunction associated with experimental cirrhosis . We examined the functional role of nitric oxide ( NO ) and nitric oxide synthase ( NOS ) isoforms in the pulmonary dysfunction seen in cirrhosis . Lungs were isolated from control and carbon tetrachloride ( CCl(4) ) -induced cirrhotic rats and perfused at constant flow with a whole blood mixture . Ventilation with hypoxic gas resulted in attenuated hypoxic pulmonary vasoconstriction ( HPV ) in lungs from cirrhotic animals . Administration of the non-selective NOS inhibitor N-omega-Nitro-L- DB00125 ( DB04223 ) resulted in HPV responses that were not different between groups . However , inhibition of inducible nitric oxide synthase ( P35228 ) did not restore cirrhotic HPV responses . Lungs from cirrhotic rats demonstrated enhanced endothelial-dependent vasodilation to vasopressin when preconstricted with hypoxia but not when preconstricted with thromboxane mimetic . Western blot analysis failed to demonstrate differences in pulmonary endothelial NOS ( P29474 ) or P35228 levels between groups . Our data suggest that , while NO may play a role in mediating the reduced pulmonary vasoreactivity observed in cirrhosis , other vasoactive factors are likely also important modulators of the pulmonary dysfunction seen in this disease . Phosphodiesterase-4 gates the ability of protein kinase A to phosphorylate G-protein receptor kinase-2 and influence its translocation . Challenge of the beta(2)Ar ( beta(2)-adrenergic receptor ) with isoprenaline in P29320 -293beta(2) cells ( human embryonic kidney cells stably overexpressing a FLAG- and green fluorescent protein-tagged beta(2)Ar ) results in the PKA ( DB02527 -dependent protein kinase ) phosphorylation of P25098 ( G-protein receptor kinase-2 ) . This response was enhanced when DB05876 ( phosphodiesterase-4 ) activity was attenuated using either rolipram , a DB05876 -selective inhibitor , or with siRNA ( small interfering RNA ) knockdown of both Q07343 and Q08499 . DB01954 also facilitated P25098 recruitment to the membrane and phosphorylation of the beta(2)Ar by P25098 in response to isoprenaline challenge of cells . In resting cells , rolipram treatment alone is sufficient to promote PKA phosphorylation of P25098 , with consequential effects on P25098 translocation and P25098 phosphorylation of the beta(2)Ar . Similar effects are observed in cardiac myocytes . We propose that DB05876 activity protects P25098 from inappropriate phosphorylation by PKA in resting cells that might have occurred through fluctuations in basal DB02527 levels . Thus DB05876 gates the action of PKA to phosphorylate P25098 . 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 . Enhanced NF-κB activity impairs vascular function through P09874 - , SP-1- , and P35354 -dependent mechanisms in type 2 diabetes . Type 2 diabetes ( T2D ) is associated with vascular dysfunction . We hypothesized that increased nuclear factor-κB ( NF-κB ) signaling contributes to vascular dysfunction in T2D . We treated type 2 diabetic ( db(-)/db(-) ) and control ( db(-)/db(+) ) mice with two NF-κB inhibitors ( 6 mg/kg dehydroxymethylepoxyquinomicin twice a week and 500 μg/kg/day IKK-NBD peptide ) for 4 weeks . Pressure-induced myogenic tone was significantly potentiated , while endothelium-dependent relaxation ( EDR ) was impaired in small coronary arterioles and mesenteric resistance artery from diabetic mice compared with controls . Interestingly , diabetic mice treated with NF-κB inhibitors had significantly reduced myogenic tone potentiation and improved EDR . Importantly , vascular function was also rescued in db(-)/db(-p50NF-κB-/-) and db(-)/db(- P09874 -/-) double knockout mice compared with db(-)/db(-) mice . Additionally , the acute in vitro downregulation of NF-κB-p65 using p65NF-κB short hairpin RNA lentivirus in arteries from db(-)/db(-) mice also improved vascular function . The NF-κB inhibition did not affect blood glucose level or body weight . The RNA levels for Sp-1 and P29474 phosphorylation were decreased , while p65NF-κB phosphorylation , cleaved poly(ADP-ribose) polymerase ( PARP ) -1 , and cyclooxygenase ( P36551 ) -2 expression were increased in arteries from diabetic mice , which were restored after NF-κB inhibition and in db(-)/db(-p50NF-κB-/-) and db(-)/db(- P09874 -/-) mice . In the current study , we provided evidence that enhanced NF-κB activity impairs vascular function by P09874 - , Sp-1- , and P35354 -dependent mechanisms in male type 2 diabetic mice . Therefore , NF-κB could be a potential target to overcome diabetes-induced vascular dysfunction . Effects of DB09045 on Thyroid C Cells and Serum P01258 in Male Monkeys . Glucagon-like peptide-1 ( P0C6A0 ) receptor agonists , used for the treatment of type 2 diabetes , have caused hyperplasia/neoplasia of thyroid C cells in rodent carcinogenicity studies . Studies in monkeys have not identified an effect of P43220 agonists on thyroid C cells ; however , group sizes were small . DB09045 is a once-weekly , long-acting human P43220 agonist recently approved in the United States and the European Union . The objective of this study was to determine whether dulaglutide altered C-cell mass in monkeys . Male cynomolgus monkeys ( 20 per group ) were sc injected with dulaglutide 8.15 mg/kg ( ∼500-fold maximum human plasma exposure ) or a vehicle control twice weekly for 52 weeks . Basal and calcium gluconate-stimulated serum calcitonin concentrations were obtained at 3 , 6 , 9 , and 12 months . Thyroid glands were weighed , fixed , and sectioned at 500-μm intervals . C-cell volumes were measured using an automated image analysis . C-cell proliferation was estimated using Ki67/calcitonin colabeling and cell counting . Administration of dulaglutide 8.15 mg/kg twice weekly for 52 weeks did not increase serum calcitonin in monkeys or affect thyroid weight , histology , C-cell proliferation , or absolute/relative C-cell volume . This study represents a comprehensive evaluation of the monkey thyroid C cells after dosing with a P43220 agonist , with a large group size , and measurement of multiple relevant parameters . The lack of effect of dulaglutide on C cells is consistent with other studies in monkeys using P43220 agonists and suggests that nonhuman primates are less sensitive than rodents to the induction of proliferative changes in thyroid C cells by P43220 agonists . Whole blood lead concentration and erythrocyte delta-aminolevulinic acid dehydratase ( P13716 ) activity in selected canine populations in Greece . In a total number of 275 dogs of various ages , sex and breed , blood lead concentrations ( O43927 ) and erythrocyte P13716 activity were measured . Sixty-six of the dogs were living in lead mining areas ( Group A ) , 157 in urban areas ( Group B ) and 52 in rural areas ( Group C ) of Greece . Mean O43927 differed significantly ( P < 0.05 ) between locations and were 326,97 and 68 micrograms/L , respectively . Mean P13716 activity was significantly different ( P < 0.05 ) only between Groups A and B as between groups A and C . A significant ( P < 0.05 ) negative correlation existed between O43927 and P13716 activity . A normal range of erythrocyte P13716 activity of 807-992 mumol/ DB02272 /LRBC/h was established for dogs . None of the 33 Group A dogs and 2 of the Group B dogs that had a O43927 of 350 micrograms/L presented clinical signs indicating acute or chronic lead intoxication . No erythrocyte basophilic stippling or large number of nucleated red blood cells were seen in the 30 dogs of Group A with O43927 > 350 micrograms/L .

MH_train_1197

MH_train_1197

MH_train_1197

interacts_with DB00603?

multiple_choice

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 . 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 . 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 . EDRF suppresses an unidentified vasoconstrictor mechanism in hypertensive rat lungs . To test whether DB00435 ( EDRF ) plays a role in regulating the hypertensive pulmonary vascular bed , we compared effects of the inhibitor of EDRF production , N omega-nitro-L-arginine ( DB04223 ) , on resting vascular tone in lungs and conduit pulmonary arteries isolated from control and chronically hypoxic rats . In contrast to no effect on normoxic vascular tone in salt solution-perfused normotensive lungs , 100 microM DB04223 caused a marked , L-arginine-sensitive , precapillary vasoconstriction in unstimulated hypertensive lungs . Bioassay of hypertensive lung perfusate did not detect a circulating vasoconstrictor , and DB04223 vasoconstriction was not inhibited by blockers of cyclooxygenase , P09917 , platelet-activating factor receptors , alpha-adrenoceptors , and serotonin 5-HT2 receptors or by scavengers of superoxide anion and H2O2 . Inhibitors of endothelin-1 ( ET-1 ) production and vasoconstriction tended to blunt the response , but accumulation of perfusate ET-1 was not increased in hypertensive lungs . DB04223 vasoconstriction was blocked by Ca(2+)-free plus ethylene glycol-bis ( beta-aminoethyl ether ) -N,N,N',N'-tetraacetic acid perfusion but not by nifedipine . Quiescent , endothelium-intact hypertensive but not normotensive conduit pulmonary artery rings were markedly constricted by 200 microM DB04223 . The onset but not the peak of the response was blunted by meclofenamate . The response was reduced slightly by the P25101 receptor antagonist , BQ 123 . DB04223 had little effect on denuded hypertensive arteries , and treatment with dilators showed they had constricted spontaneously . Both the DB04223 and the spontaneous constrictions were readily inhibited by nifedipine . These results indicate that in rat hypertensive pulmonary arteries , the basal release of EDRF suppresses vasoconstrictor mechanisms which are not expressed in normotensive arteries . Genomic analyses and cloning of novel chicken natriuretic peptide genes reveal new insights into natriuretic peptide evolution . The natriuretic peptide ( NP ) family consists of multiple subtypes in teleosts , including atrial , B-type , ventricular , and C-type NPs ( P01160 , DB04899 , VNP , P09543 -1-4 , respectively ) , but only P01160 , DB04899 , P09543 -3 , and P09543 -4 have been identified in tetrapods . As part of understanding the molecular evolution of NPs in the tetrapod lineage , we identified NP genes in the chicken genome . Previously , only DB04899 and P09543 -3 have been identified in birds , but we characterized two new chicken NP genes by cDNA cloning , synteny and phylogenetic analyses . One gene is an orthologue of P09543 -1 , which has only ever been reported in teleostei and bichir . The second gene could not be assigned to a particular NP subtype because of high sequence divergence and was named renal NP ( Q96LT9 ) due to its predominant expression in the kidney . P09543 -1 mRNA was only detected in brain , while P09543 -3 mRNA was expressed in kidney , heart , and brain . In the developing embryo , DB04899 and Q96LT9 transcripts were most abundant 24h post-fertilization , while P09543 mRNA increased in a stage-dependent manner . Synthetic chicken Q96LT9 stimulated an increase in cGMP production above basal level in chicken kidney membrane preparations and caused a potent dose-dependent vasodilation of pre-constricted dorsal aortic rings . From conserved chromosomal synteny , we propose that the P09543 -4 and P01160 genes have been lost in chicken , and that Q96LT9 may have evolved from a VNP-like gene . Furthermore , we have demonstrated for the first time that P09543 -1 is retained in the tetrapod lineage . 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 . Biological characterization of DB05037 , a small-molecule inhibitor of cyclin-dependent kinases , in human tumor cell lines . P12004 -dependent kinases ( CDK ) , and their regulatory cyclin partners , play a central role in eukaryotic cell growth , division , and death . This key role in cell cycle progression , as well as their deregulation in several human cancers , makes them attractive therapeutic targets in oncology . A series of CDK inhibitors was developed using Astex 's fragment-based medicinal chemistry approach , linked to high-throughput X-ray crystallography . A compound from this series , designated DB05037 , is currently in early-phase clinical development . We describe here the biological characterization of DB05037 , a potent inhibitor of several CDK family members . DB05037 showed potent antiproliferative activity ( 40-940 nmol/L ) in a panel of human tumor cell lines , and the mechanism of action was shown here to be consistent with the inhibition of P06493 and P24941 in solid tumor cell lines . DB05037 caused cell cycle arrest followed by apoptosis in human tumor cells and inhibited tumor growth in human tumor xenograft models . Tumor regression was observed following twice daily dosing of DB05037 in the HCT116 and HT29 colon cancer xenograft models . We show that these biological effects are linked to inhibition of CDKs in vivo and that DB05037 induces tumor cell apoptosis in these xenograft models . DB05037 has an attractive biological profile for development as a clinical candidate , and the tolerability and efficacy in animal models compare favorably with other CDK inhibitors in clinical development . Studies described here formed the biological rationale for investigating the potential therapeutic benefit of DB05037 in cancer patients . Endothelial nitric-oxide synthase activation generates an inducible nitric-oxide synthase-like output of nitric oxide in inflamed endothelium . High levels of NO generated in the vasculature under inflammatory conditions are usually attributed to inducible nitric-oxide synthase ( P35228 ) , but the role of the constitutively expressed endothelial NOS ( P29474 ) is unclear . In normal human lung microvascular endothelial cells ( HLMVEC ) , bradykinin ( BK ) activates kinin B2 receptor ( P30411 ) signaling that results in Ca(2+)-dependent activation of P29474 and transient NO . In inflamed HLMVEC ( pretreated with interleukin-1β and interferon-γ ) , we found enhanced binding of P29474 to calcium-calmodulin at basal Ca(2+) levels , thereby increasing its basal activity that was dependent on extracellular l- DB00125 . Furthermore , P30411 stimulation generated prolonged high output P29474 -derived NO that is independent of increased intracellular Ca(2+) and is mediated by a novel Gα(i)- , Q02750 /2- , and P45983 /2-dependent pathway . This high output NO stimulated with BK was blocked with a P30411 antagonist , P29474 siRNA , or P29474 inhibitor but not P35228 inhibitor . Moreover , P30411 -mediated NO production and JNK phosphorylation were inhibited with Q02750 /2 and JNK inhibitors or Q02750 /2 and P45983 /2 siRNA but not with P27361 /2 inhibitor . BK induced Ca(2+)-dependent P29474 phosphorylation at DB00133 (1177) , DB00156 (495) , and DB00133 (114) in cytokine-treated HLMVEC , but these modifications were not dependent on P45983 /2 activation and were not responsible for prolonged NO output . Cytokine treatment did not alter the expression of P30411 , Gα(q/11) , Gα(i1,2) , JNK , or P29474 . P30411 activation in control endothelial cells enhanced migration , but in cytokine-treated HLMVEC it reduced migration . Both responses were NO-dependent . Understanding how JNK regulates prolonged P29474 -derived NO may provide new therapeutic targets for the treatment of disorders involving vascular inflammation . Concentrations of N-terminal ProANP in human plasma : evidence for ProANP ( 1-98 ) as the circulating form . Plasma levels of immunoreactive N-terminal ProANP have been measured in plasma from 19 healthy individuals , 15 patients with essential hypertension , 8 cardiac transplant recipients and 8 patients with chronic renal failure using two separate radioimmunoassays ( RIAs ) , one directed against ProANP ( 1-30 ) and the other against ProANP ( 79-98 ) . The mean concentrations of ProANP ( 1-30 ) and ProANP ( 79-98 ) were elevated in these groups of patients . There were positive correlations between levels of ProANP ( 1-30 ) and ProANP ( 79-98 ) , with a correlation coefficient of 0.97 ( P less than 0.001 , n = 50 ) . In healthy individuals a P35326 ( isotonic ) saline infusion significantly increased both P01160 ( 99-126 ) ( P less than 0.05 , n = 8 ) and N-terminal ProANP ( P less than 0.005 , n = 8 ) within 15 min of the end of the infusion . Plasma N-terminal ProANP levels were still significantly elevated after 75 min ( P less than 0.05 , n = 8 ) and 225 min ( P less than 0.05 , n = 8 ) , by contrast P01160 ( 99-126 ) had returned to basal values . Gel filtration of plasma extracted on Sep-Pak C-18 from normal individuals and patients gave a single immunoreactive peak for N-terminal ProANP as measured by both N-terminal ProANP assays , indicating an absence of small N-terminal fragments and the presence of a single high molecular weight form . These studies demonstrate that the major circulating N-terminal P01160 in man is probably ProANP ( 1-98 ) and that it is cosecreted with P01160 ( 99-126 ) . Specific phosphorylation of SR proteins by mammalian P11387 . Several metazoan splicing factors are characterized by ribonucleoprotein ( Q96LT9 ) consensus sequences and arginine-serine repeats ( RS domain ) which are essential for their function in splicing . These include members of the SR-protein family ( SC35 , Q07955 /ASF ) , the U1 small nuclear ( sn ) Q96LT9 protein ( P08621 ) and the U2 snRNP auxiliary factor ( U2AF ) . SR proteins are phosphorylated in vivo and the phosphorylation state of P08621 's RS domain influences its splicing activity . Here we report the purification of a protein kinase that is specific for SR proteins and show that it is P11387 . This enzyme lacks a canonical DB00171 -binding motif but binds DB00171 with a dissociation constant of 50 nM . DB04690 and derivatives , known to be specific inhibitors of P11387 , strongly inhibit the kinase activity in the presence of DNA and affect the phosphorylation state of SR proteins . Thus , P11387 may well be one of the SR protein kinases operating in vivo . 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 . Expression and biological activity of O95477 in alveolar epithelial cells . The mechanisms used by alveolar type I pneumocytes for maintenance of the lipid homeostasis necessary to sustain these large squamous cells are unknown . The processes may involve the DB00171 -binding cassette transporter A1 ( O95477 ) , a transport protein shown to be crucial in apolipoprotein A-I ( apoA-I ) -mediated mobilization of cellular cholesterol and phospholipid . Immunohistochemical data demonstrated the presence of O95477 in lung type I and type II cells and in cultured pneumocytes . Type II cells isolated from rat lungs and cultured for 5 days in 10 % serum trans-differentiated toward cells with a type I-like phenotype which reacted with the type I cell-specific monoclonal antibody VIIIB2 . Upon incubation of the type I-like pneumocytes with agents that up-regulate the O95477 gene ( 9-cis-retinoic acid [ 9cRA ] and 22-hydroxycholesterol [ 22-OH , 9cRA/22-OH ] ) , O95477 protein levels were enhanced to maximum levels after 8 to 16 hours and remained elevated for 24 hours . In the presence of apoA-I and 9cRA/22-OH , efflux of radioactive phospholipid and cholesterol from pneumocytes was stimulated 3- to 20-fold , respectively , over controls . Lipid efflux was inhibited by DB01599 . DB02772 density gradient analysis of the media from stimulated cells incubated with apoA-I identified heterogeneous lipid particles that isolated at a density between 1.063 and 1.210 g/ml , with low or high apoA-I content . Thus , pneumocytes with markers for the type I phenotype contained functional O95477 protein , released lipid to apoA-I protein , and were capable of producing particles resembling nascent high-density lipoprotein , indicating an important role for O95477 in the maintenance of lung lipid homeostasis . Leukotriene A4 hydrolase . Inhibition by bestatin and intrinsic aminopeptidase activity establish its functional resemblance to metallohydrolase enzymes . DB03424 , an inhibitor of aminopeptidases , was also a potent inhibitor of leukotriene ( LT ) A4 hydrolase . On isolated enzyme its effects were immediate and reversible with a Ki = 201 +/- 95 mM . With erythrocytes it inhibited LTB4 formation greater than 90 % within 10 min ; with neutrophils it inhibited LTB4 formation by only 10 % during the same period , increasing to 40 % in 2 h . DB03424 inhibited P09960 hydrolase selectively ; neither P09917 nor 15-lipoxygenase activity in neutrophil lysates was affected . Purified P09960 hydrolase exhibited an intrinsic aminopeptidase activity , hydrolyzing L-lysine-p-nitroanilide and L-leucine-beta-naphthylamide with apparent Km = 156 microM and 70 microM and Vmax = 50 and 215 nmol/min/mg , respectively . Both P09960 and bestatin suppressed the intrinsic aminopeptidase activity of P09960 hydrolase with apparent Ki values of 5.3 microM and 172 nM , respectively . Other metallohydrolase inhibitors tested did not reduce P09960 hydrolase/aminopeptidase activity , with one exception ; captopril , an inhibitor of angiotensin-converting enzyme , was as effective as bestatin . The results demonstrate a functional resemblance between P09960 hydrolase and certain metallohydrolases , consistent with a molecular resemblance at their putative Zn2(+)-binding sites . The availability of a reversible , chemically stable inhibitor of P09960 hydrolase may facilitate investigations on the role of LTB4 in inflammation , particularly the process termed transcellular biosynthesis . Differential effects of protein kinase C , Ras , and P04049 kinase on the induction of the cardiac B-type natriuretic peptide gene through a critical promoter-proximal M-CAT element . The cardiac genes for the A- and B-type natriuretic peptides ( P01160 and DB04899 ) are coordinately induced by growth promoters , such as alpha1-adrenergic receptor agonists ( e.g. phenylephrine ( PE ) ) . Although inducible elements in the P01160 gene have been identified , responsible elements in the DB04899 gene are unknown . In this study , reporter constructs transfected into neonatal rat ventricular myocytes showed that in the context of 2.5 kilobase pairs of native DB04899 5'-flanking sequences , a 2-base pair mutation in a promoter-proximal M-CAT site ( CATTCT ) disrupted basal and PE-inducible transcription by more than 98 % . Expression of constitutively active forms of Ras , P04049 kinase , and protein kinase C , all of which are activated by PE in cardiac myocytes , strongly stimulated DB04899 reporter expression . Isolated M-CAT elements conferred PE , protein kinase C , and Ras inducibility to a minimal DB04899 promoter , however , they did not confer P04049 inducibility . These results show that M-CAT elements can serve as targets for Ras-dependent , P04049 -independent pathways , implying the involvement of c-Jun N-terminal kinase and/or p38 mitogen-activated protein kinases , but not extracellular signal-regulated protein kinase/mitogen-activated protein kinase . Moreover , the essential M-CAT element distinguishes the DB04899 gene from the P01160 gene , which utilizes serum response elements and an Sp1-like sequence . Clinical development of eniluracil : current status . DB03516 is a potent inactivator of dihydropyrimidine dehydrogenase ( Q12882 ) , which is the first enzyme in the degradative pathway of systemically administered 5-fluorouracil ( DB00544 ) . Two completely oral regimens of eniluracil plus DB00544 are being evaluated in clinical trials : ( 1 ) a chronic schedule with both agents administered P55957 in a 10:1 ratio for 28 days of a 5-week course , and ( 2 ) a 5-day schedule of eniluracil once daily on days 1 through 7 and DB00544 once daily on days 2 through 6 . The clinical development of eniluracil is being pursued in several tumor types , including colorectal cancer , breast cancer , and pancreatic cancer . Response rates achieved in a phase II study of the chronic schedule of oral eniluracil/ DB00544 in patients with colorectal cancer compare favorably with those obtained in trials of intravenous DB00544 and leucovorin , while results from other trials are awaited . Safety analysis for the 28-day schedule has revealed a low incidence of severe toxicities , particularly as compared with standard DB00544 regimens . Downregulation of liver X receptor-alpha in mouse kidney and HK-2 proximal tubular cells by LPS and cytokines . The acute-phase response ( Q07954 ) suppresses type II nuclear hormone receptors and alters the expression of their target genes involved in lipid metabolism in the liver and heart . Therefore , we examined the expression of liver X receptor/retinoid X receptor ( LXR/RXR ) and their target genes in kidney from mice treated with lipopolysaccharide ( LPS ) and in human proximal tubular HK-2 cells treated with interleukin-1beta ( IL-1beta ) and tumor necrosis factor-alpha ( P01375 ) . We found that LXRalpha and RXRalpha expression was suppressed by LPS in kidney and by IL-1beta or P01375 in HK-2 cells . The decrease in LXRalpha/RXRalpha expression was associated with a decrease in the expression of several LXRalpha target genes [ apolipoprotein E ( apoE ) , O95477 , P45844 , and sterol-regulatory element binding protein-1c ( SREBP-1c ) ] and a decrease in ligand-induced apoE expression . Moreover , IL-1beta and P01375 significantly reduced liver X receptor response element ( LXRE ) -driven transcription as measured by LXRE-linked luciferase activity . However , overexpression of LXRalpha/RXRalpha only partially restored the cytokine-mediated reduction in LXRE-linked luciferase activity . Additionally , expression of the LXR coactivators peroxisome proliferator-activated receptor gamma coactivator 1alpha ( PGC1alpha ) and steroid receptor coactivator-2 ( P12931 -2 ) was decreased by IL-1beta or P01375 . We conclude that the Q07954 suppresses the expression of both nuclear receptors LXRalpha/RXRalpha and several LXRalpha coactivators in kidney , which could be a mechanism for coordinately regulating the expression of multiple LXR target genes that play important roles in lipid metabolism in kidney during the Q07954 . In utero exposure to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin affects the development of reproductive system in mouse . PURPOSE : Exposure of male reproductive organs to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin ( TCDD ) has been reported to cause developmental changes . In this study , we evaluated the effects of in utero TCDD exposure on male reproductive development . MATERIALS AND METHODS : Pregnant C57BL/6 mice were administered a single intraperitoneal injection of TCDD ( 1 microg/kg ) on gestation day ( GD ) 15 . The offspring were examined in the immature stage on postnatal day ( P01160 ) 30 and in the mature stage on P01160 60 . The testes were examined for histological changes , androgen receptor ( AR ) , proliferating cell nuclear antigen ( P12004 ) and apoptosis following the measurement of morphological changes . RESULTS : Anogenital distance ( AGD ) and testis weights were reduced by TCDD exposure both on P01160 30 and P01160 60 while body weights and length of male offspring were not affected by TCDD . The regular sperm developmental stage was impaired with TCDD treatment on P01160 30 . However , no difference was found between the control group and TCDD groups on P01160 60 . Simultaneously , the expression of AR was also reduced on P01160 30 , while it was increased on P01160 60 compared with the control group . The expression of P12004 was decreased whereas apoptosis was not affected by TCDD both on P01160 30 and P01160 60 . CONCLUSION : These results suggest that in utero exposure to TCDD influences the development of testes by inhibiting the expression of AR and P12004 . Moreover , the adverse effects of TCDD on male offspring reduced over time . Enhanced therapeutic effects of doxorubicin and paclitaxel in combination with liposome-entrapped ends-modified raf antisense oligonucleotide against human prostate , lung and breast tumor models . P04049 protein kinase plays an important role in cell growth , proliferation and cell survival . We have previously described the use of liposome-entrapped antisense raf oligonucleotide ( DB04973 ) to inhibit P04049 expression resulting in tumor growth inhibition and radiosensitization . The present study was undertaken to evaluate the chemosensitization effects of DB04973 in combination with doxorubicin or paclitaxel on a panel of human tumor xenografts . DB04973 ( 25.0 mg/kg i.v. x 10 ) displayed significant antitumor activity ( P < 0.05 ) when administered as a single agent in prostate ( PC-3 ) , lung ( A549 ) and breast ( MDA-MB 231 ) carcinoma models . Doxorubicin ( 1.0-4.0 mg/kg i.v. per week x 3 ) and paclitaxel ( 1.0-4.0 mg/kg i.v. on alternate days x 3 ) were administered as single agents at non-toxic doses that led to only minimal to moderate antitumor activity . However , a combination of DB04973 with doxorubicin or paclitaxel led to significantly enhanced antitumor activity in all the tumor types tested ( PC-3 , P < 0.03 ; A549 , P < 0.035 ; MDA-MB 231 , P < 0.045 ) as compared with DB04973 alone or chemotherapeutic agents alone treated groups . This effect of chemosensitization appeared to be sequence-specific because a mismatch control oligonucleotide continued to show significant tumor growth . Additionally , no inhibition in P04049 expression in MDA-MB 231 tumor tissue was observed with mismatch oligonucleotide treatment . On the other hand , DB04973 treatment led to > 75 % inhibition of P04049 expression in tumor tissue . These preclinical observations support the use of DB04973 in combination with chemotherapeutic agents to improve the treatment of human cancers . 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 . Population pharmacokinetics of ethionamide in patients with tuberculosis . SETTING : Three US referral hospitals . OBJECTIVE : Determine the population pharmacokinetic ( PK ) parameters of ethionamide ( P25101 ) following multiple oral doses . DESIGN : Fifty-five patients with tuberculosis ( TB ) participated . Patients received multiple oral doses of P25101 as part of their treatment . They also received other anti-tuberculosis medications based upon in vitro susceptibility data . Serum samples were collected over 12 h post-dose , and concentrations were determined using a validated high-performance liquid chromatography ( HPLC ) assay . Concentration-time data were analyzed using population methods . RESULTS : P25101 areas under the concentration-versus-time curve ( AUCs ) increased linearly with increasing oral doses from 250 to 1000 mg . Compared to the population pattern , delayed absorption was seen at least once in 15 % of patients . P25101 PK parameter estimates were independent of age , weight , height , gender , and creatinine clearance . TB patients appeared to have larger volumes of distribution ( 3.22 l/kg ) and clearance values ( 1.88 l/h/kg ) compared to previously studied healthy volunteers . This resulted in lower AUC values ( 3.95 mcg h/ml ) in the TB patients . P25101 displayed a short elimination half-life ( 1.94 h ) . The effect of different dosing strategies on calculated pharmacodynamic parameters was explored . Simulated doses of 250 mg P55957 to TID failed to achieve serum concentrations above the MIC . CONCLUSION : P25101 PK parameters differed between TB patients and healthy volunteers , possibly due to differences in the completeness of absorption . Doses of at least 500 mg appear to be required to achieve serum concentrations above the typical P25101 MIC . Additional research is needed to determine the optimal dosing of P25101 . P06401 modulator for emergency contraception : a randomized controlled trial . OBJECTIVE : Compare the efficacy and adverse effects of DB05366 , a new progesterone receptor modulator , to levonorgestrel for emergency contraception . METHODS : We performed a randomized , double-blinded noninferiority trial , enrolling healthy women seeking emergency contraception within 72 hours of unprotected intercourse . Participants were randomly assigned to receive a single dose of 50 mg of DB05366 , plus a placebo 12 hours later or two doses of 0.75 mg of levonorgestrel taken 12 hours apart . Follow-up was scheduled 5 to 7 days after the expected onset of the next menstrual period . Posttreatment pregnancy was established by a positive urine test at follow-up and confirmed by quantitative serum beta-hCG . Daily diaries were used from the time of emergency contraception use until next menses to record adverse effects and sexual activity . RESULTS : Product efficacy was evaluable in 775 of DB05366 users and 774 of levonorgestrel users . Pregnancies occurred in 7 ( 0.9 % , 95 % confidence interval 0. P35326 .6 % ) and 13 ( 1.7 % , 95 % confidence interval 0.8-2.6 % ) women , respectively . Based on the estimated cycle day of unprotected intercourse , 85 % and 69 % of anticipated pregnancies , respectively , were averted . Nausea was reported by a somewhat greater percentage of DB05366 than levonorgestrel users ( 29 % compared with 24 % , P=.03 ) , but the distribution of other adverse effects was similar in both groups . Women in both groups experienced considerable variation in menstrual cycle length as compared with their reported individual normal cycle lengths . CONCLUSION : DB05366 is at least as effective as levonorgestrel in preventing pregnancies after unprotected intercourse and has a similar side effect profile . LEVEL OF EVIDENCE : I .

MH_train_1198

MH_train_1198

MH_train_1198

interacts_with DB00215?

multiple_choice

Regulation of Con A-dependent cytokine production from P01730 + and CD8+ T lymphocytes by autosecretion of histamine . OBJECTIVES : Previously we have shown that both P01730 + T cells and CD8+ T cells produce histamine when activated with Con A . The aim of this study was to examine whether cytokine production by these cells is regulated by autosecretion of histamine . MATERIALS : P01730 + and CD8+ T cells were separated from spleen cells of C57BL/6 mice and mice lacking the H1 receptor ( P35367 ) or P25021 , using anti- P01730 +- and anti-CD8+-coupled magnetic beads , respectively . RESULTS : Depletion of the P35367 resulted in decreases in the release of P60568 and P22301 from both P01730 + and CD8+ cells and increases in the release of P05112 from P01730 + T cells and P01579 from CD8+ cells . Mice lacking the P25021 showed up-regulation of P01579 secretion from CD8+ cells and of P05112 from P01730 + and CD8+ T cells . Release of P60568 and P22301 from P01730 + as well as CD8+ cells was down-regulated in these mice . Both P01730 + and CD8+ T cell fractions synthesized histamine , which was enhanced in the P35367 -deficient CD8+ T cells . Treatment of the cells with alpha-fluoromethyl-histidine , a specific inhibitor of HDC , or histaminase increased P01579 from CD8+ cells , whereas it had no appreciable effect on P05112 secretion from P01730 + cells . CONCLUSION : These results suggest that cytokine production by P01730 + and CD8+ T lymphocytes is regulated by autosecretion of histamine . 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 . [ Regulation of cell growth exemplified by T lymphocytes ] . The aim of this paper is to describe the complexicity of growth regulation , particular of T-lymphocytes . Different surface molecules are known , which trigger the process of cell activation after binding of the respective ligands , including the T-cell-receptor/CD3 complex , the P06729 and the P01730 molecules . This surface molecules induce among other cellular events the expression of additional receptors , such as P60568 -receptors and P02787 -receptors . The structure of these surface molecules , their possible functions and involved mechanisms of signal transmission from the membrane to the level of gene expression are presented . The different mechanisms of signal transmission and structures involved , such as the G-protein , are discussed . The different systems of signal transmission , in which such enzymes like Ca2+-dependent neutral Protease , Phospholipase C , Adenylatcyclase , P04054 and Proteinkinases are involved , can interact . The regulation of the proliferation of T-cells is the result of a common action of different lymphokines , but also of growth factors from non-immune cells . 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 . 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 . Pharmacological profile of astemizole-derived compounds at the histamine H1 and H4 receptor -- H1/H4 receptor selectivity . DB00637 , a P35367 antagonist shows high affinity to the histamine H1 receptor but only a moderate affinity to the histamine H4 receptor . This study aims to modify the astemizole to keep high affinity to the histamine H1 receptor and to increase affinity to the histamine H4 receptor . Therefore , 13 astemizole-derived compounds and astemizole-JNJ7777120-derived hybrid compounds were synthesized and pharmacologically characterized at the histamine H1 and H4 receptors . The new compounds show affinity to the histamine H1 receptor in the pK i range from 5.3 to 8.8 , whereas the affinity of these compounds to the histamine H4 receptor was surprisingly rather low ( pK i from 4.4 to 5.6 ) . Three representative compounds were docked into the histamine H1 receptor and molecular dynamic studies were performed to explain the binding mode and the experimental results on a molecular level . Furthermore , taking into account the binding mode of compounds with high affinity to the histamine H4 receptor , a H1/H4-pharmacophore hypothesis was developed . 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 . Selective P36888 inhibition of P36888 -ITD+ acute myeloid leukaemia resulting in secondary D835Y mutation : a model for emerging clinical resistance patterns . Acquired resistance to selective P36888 inhibitors is an emerging clinical problem in the treatment of P36888 -ITD(+) acute myeloid leukaemia ( AML ) . The paucity of valid pre-clinical models has restricted investigations to determine the mechanism of acquired therapeutic resistance , thereby limiting the development of effective treatments . We generated selective P36888 inhibitor-resistant cells by treating the P36888 -ITD(+) human AML cell line MOLM-13 in vitro with the P36888 -selective inhibitor MLN518 , and validated the resistant phenotype in vivo and in vitro . The resistant cells , MOLM-13-RES , harboured a new D835Y tyrosine kinase domain ( TKD ) mutation on the P36888 -ITD(+) allele . Acquired TKD mutations , including D835Y , have recently been identified in P36888 -ITD(+) patients relapsing after treatment with the novel P36888 inhibitor , DB05213 . Consistent with this clinical pattern of resistance , MOLM-13-RES cells displayed high relative resistance to DB05213 and DB00398 . Furthermore , treatment of MOLM-13-RES cells with DB05213 lead to loss of the P36888 wild-type allele and the duplication of the P36888 -ITD-D835Y allele . Our P36888 -Aurora kinase inhibitor , CCT137690 , successfully inhibited growth of P36888 -ITD-D835Y cells in vitro and in vivo , suggesting that dual P36888 -Aurora inhibition may overcome selective P36888 inhibitor resistance , in part due to inhibition of Aurora kinase , and may benefit patients with P36888 -mutated AML . Inhibition of SKF 38393- and pergolide-induced circling in rats with unilateral 6-OHDA lesion is correlated to dopamine D-1 and D-2 receptor affinities in vitro . The effects of 30 dopamine ( DA ) antagonists , including 4 as stereoisomeric pairs , on circling behaviour induced by the D-1 agonist SKF 38393 and the D-2 agonist pergolide in rats with unilateral 6-hydroxy-DA lesions have been studied . SKF 38393-induced circling was selectively blocked by the specific D-1 antagonists P35240 23390 and SKF 83566 , and was furthermore blocked by other DA antagonists with potencies correlating to their affinities to D-1 receptors labelled by 3H- P35240 23390 in vitro . DB01186 -antagonistic potencies in contrast correlated to affinities to D-2 receptors labelled by 3H-spiperone in vitro . DB01186 -induced circling was selectively blocked by the specific D-2 antagonists in the benzamide series . No interaction between D-1 and D-2 antagonists was observed in combination experiments with P35240 23390 and YM 09151-2 in both circling models . Among other reference neurotransmitter antagonists acting on alpha- and beta-adrenoceptors , histamine , serotonin and muscarinic receptors , only the alpha 1-adrenoceptor antagonist prazosin was effective in high doses . In contrast , the alpha 2- and beta-adrenoceptor agonists clonidine and clenbuterol as well as the muscarinic agonist RS 86 inhibited circling induced by SKF 38393 as well as pergolide . The P08908 agonist 8-OHDPAT inhibited pergolide-induced circling only . It is concluded that these two behavioural models are selective in vivo measures of relative D-1 and D-2 receptor activity of DA antagonists . Enhancement of auranofin-induced apoptosis in MCF-7 human breast cells by selenocystine , a synergistic inhibitor of thioredoxin reductase . P10599 system plays an important role in regulation of intracellular redox balance and various signaling pathways . P30044 ( TrxR ) is overexpressed in many cancer cells and has been identified as a potential target of anticancer drugs . DB00995 ( AF ) is potent TrxR inhibitor with novel in vitro and in vivo anticancer activities . Selenocystine ( SeC ) is a nutritionally available selenoamino acid with selective anticancer effects through induction of apoptosis . In the present study , we demonstrated the synergistic effects and the underlying molecular mechanisms of SeC in combination with AF on MCF-7 human breast cancer cells . The results showed that SeC and AF synergistically inhibited the cancer cell growth through induction of ROS-dependent apoptosis with the involvement of mitochondrial dysfunction . DNA damage-mediated p53 phosphorylation and down-regulation of phosphorylated AKT and P29323 also contributed to cell apoptosis . Moreover , we demonstrated the important role of TrxR activity in the synergistic action of SeC and AF . Taken together , our results suggest the strategy to use SeC and AF in combination could be a highly efficient way to achieve anticancer synergism by targeting TrxR . Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma . Tumor-derived exosomes contain biologically active proteins and messenger and microRNAs ( miRNAs ) . These particles serve as vehicles of intercellular communication and are emerging mediators of tumorigenesis and immune escape . Here , we isolated 30-100 nm exosomes from the serum of patients with nasopharyngeal carcinoma ( NPC ) or the supernatant of TW03 cells . Increased circulating exosome concentrations were correlated with advanced lymphoid node stage and poor prognosis in NPC patients ( P < 0.05 ) . TW03-derived exosomes impaired T-cell function by inhibiting T-cell proliferation and Th1 and Th17 differentiation and promoting Treg induction by NPC cells in vitro . These results are associated with decreases in P29323 , P42224 , and P40763 phosphorylation and increases in P42229 phosphorylation in exosome-stimulated T-cells . TW03-derived exosomes increased the proinflammatory cytokines IL-1β , P05231 , and P22301 but decreased IFNγ , P60568 , and Q16552 release from P01730 + or CD8+ T-cells . Furthermore , five commonly over-expressed miRNAs were identified in the exosomes from patient sera or NPC cells : hsa-miR-24-3p , hsa-miR-891a , hsa-miR-106a-5p , hsa-miR-20a-5p , and hsa-miR-1908 . These over-expressed miRNA clusters down-regulated the Q9P0L2 signaling pathway to alter cell proliferation and differentiation . Overall , these observations reveal the clinical relevance and prognostic value of tumor-derived exosomes and identify a unique intercellular mechanism mediated by tumor-derived exosomes to modulate T-cell function in NPC . P35354 mediates microglial activation and secondary dopaminergic cell death in the mouse MPTP model of Parkinson 's disease . BACKGROUND : Accumulating evidence suggests that inflammation plays an important role in the progression of Parkinson 's disease ( PD ) . Among many inflammatory factors found in the PD brain , cyclooxygenase ( P36551 ) , specifically the inducible isoform , P35354 , is believed to be a critical enzyme in the inflammatory response . Induction of P35354 is also found in an experimental model of PD produced by administration of 1-methy-4-phenyl-1,2,3,6-tetrahydropyridine ( MPTP ) . METHOD : P35354 -deficient mice or C57BL/6 mice were treated with MPTP to investigate the effects of P35354 deficiency or by using various doses of valdecoxib , a specific P35354 inhibitor , which induces inhibition of P35354 on dopaminergic neuronal toxicity and locomotor activity impairment . Immunohistochemistry , stereological cell counts , immunoblotting , an automated spontaneous locomotor activity recorder and rotarod behavioral testing apparatus were used to assess microglial activation , cell loss , and behavioral impairments . RESULTS : MPTP reduced tyrosine hydroxylase ( TH ) -positive cell counts in the substantia nigra pars compacta ( SNpc ) ; total distance traveled , vertical activity , and coordination on a rotarod ; and increased microglia activation . DB00580 alleviated the microglial activation , the loss of TH-positive cells and the decrease in open field and vertical activity . P35354 deficiency attenuated MPTP-induced microglial activation , degeneration of TH-positive cells , and loss of coordination . CONCLUSION : These results indicate that reducing P35354 activity can mitigate the secondary and progressive loss of dopaminergic neurons as well as the motor deficits induced by MPTP , possibly by suppression of microglial activation in the SNpc . 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 . Type I IFN contributes to NK cell homeostasis , activation , and antitumor function . This study demonstrates that type I IFNs are an early and critical regulator of NK cell numbers , activation , and antitumor activity . Using both P17181 - and P48551 -deficient mice , as well as an P17181 -blocking Ab , we demonstrate that endogenous type I IFN is critical for controlling NK cell-mediated antitumor responses in many experimental tumor models , including protection from methylcholanthrene-induced sarcomas , resistance to the NK cell-sensitive RMA-S tumor and cytokine immunotherapy of lung metastases . Protection from RMA-S afforded by endogenous type I IFN is more potent than that of other effector molecules such as P01579 , IL-12 , Q14116 , and perforin . Furthermore , cytokine immunotherapy using IL-12 , Q14116 , or Q9HBE4 was effective in the absence of endogenous type I IFN , however the antimetastatic activity of P60568 was abrogated in P17181 -deficient mice , primarily due to a defect in P60568 -induced cytotoxic activity . This study demonstrates that endogenous type I IFN is a central mediator of NK cell antitumor responses . Inflammatory signaling compromises cell responses to interferon alpha . DB05258 ( IFNα ) is widely used for treatment of melanoma and certain other malignancies . This cytokine as well as the related IFNβ exerts potent anti-tumorigenic effects ; however , their efficacy in patients is often suboptimal . Here , we report that inflammatory signaling impedes the effects of IFNα/β . Melanoma cells can secrete pro-inflammatory cytokines that inhibit cellular responses to IFNα/β via activating the ligand-independent pathway for the phosphorylation and subsequent ubiquitination and accelerated degradation of the P17181 chain of type I IFN receptor . Catalytic activity of the p38 protein kinase was required for P17181 downregulation and inhibition of IFNα/β signaling induced by proinflammatory cytokines such as interleukin 1 ( IL-1 ) . Activation of p38 kinase inversely correlated with protein levels of P17181 in clinical melanoma specimens . Inhibition of p38 kinase augmented the inhibitory effects of IFNα/β on cell viability and growth in vitro and in vivo . The roles of inflammation and p38 protein kinase in regulating cellular responses to IFNα/β in normal and tumor cells are discussed . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . Catechol-o-methyltransferase expression and 2-methoxyestradiol affect microtubule dynamics and modify steroid receptor signaling in leiomyoma cells . CONTEXT : Development of optimal medicinal treatments of uterine leiomyomas represents a significant challenge . DB02342 ( 2ME ) is an endogenous estrogen metabolite formed by sequential action of CYP450s and catechol-O-methyltransferase ( P21964 ) . Our previous study demonstrated that 2ME is a potent antiproliferative , proapoptotic , antiangiogenic , and collagen synthesis inhibitor in human leiomyomas cells ( huLM ) . OBJECTIVES : Our objectives were to investigate whether P21964 expression , by the virtue of 2ME formation , affects the growth of huLM , and to explore the cellular and molecular mechanisms whereby P21964 expression or treatment with 2ME affect these cells . RESULTS : Our data demonstrated that E(2)-induced proliferation was less pronounced in cells over-expressing P21964 or treated with 2ME ( 500 nM ) . This effect on cell proliferation was associated with microtubules stabilization and diminution of estrogen receptor alpha ( ERalpha ) and progesterone receptor ( PR ) transcriptional activities , due to shifts in their subcellular localization and sequestration in the cytoplasm . In addition , P21964 over expression or treatment with 2ME reduced the expression of hypoxia-inducible factor -1alpha ( Q9BYW2 alpha ) and the basal level as well as P01375 -induced aromatase ( P11511 ) expression . CONCLUSIONS : P21964 over expression or treatment with 2ME stabilize microtubules , ameliorates E(2)-induced proliferation , inhibits ERalpha and PR signaling , and reduces Q9BYW2 alpha and P11511 expression in human uterine leiomyoma cells . Thus , microtubules are a candidate target for treatment of uterine leiomyomas . In addition , the naturally occurring microtubule-targeting agent 2ME represents a potential new therapeutic for uterine leiomyomas . AACR - 91st meeting . Dendritic cell vaccine strategies . Inevitably , the sessions on dendritic cells at the AACR Annual Meeting were some of the most consistently well attended . Interest has been intense for several years , largely since the technical obstacles to the routine culture of these cells and their precursors , both from animal and human sources , were removed in the early 1990s . Several important advances were presented towards further optimizing dendritic cell-based immunotherapy in general . Groups reported on improved culture conditions , as well as more efficient means of obtaining larger quantities of dendritic cells or their precursors . As expected , there were several strong reports on the beneficial bioactivity of cytokines , such as IL-12 , GM- P04141 , and P60568 . In addition , enticing work continues with Flt3-ligand and has begun with progenipoietin , a more recently identified hematopoietic growth factor . As shown in this year 's sessions , the clinical promise of tumor lysate and apoptotic bodies continues to move steadily from the bench to the clinic . Finally , although dendritic cells are excellent antigen-presenters , work to determine if they could be engineered to be even more effective continues . As a result , several reports were given on gene-modifying this type of immune cell . DB01370 uptake and effects on transferrin mediated iron uptake in primary cultures of rat neurons , astrocytes and oligodendrocytes . P02787 ( Tf ) is known primarily for its role in the transport and cellular uptake of iron ( Fe ) . Tf is also the major serum binding protein for Al . In this study , primary rat oligodendrocyte , neuron and astrocyte cultures were found to differ in Tf mediated Fe and Al uptake and in the effect of Al-Tf on Fe-Tf uptake during 4 h incubation periods . When incubated with Al-Tf ( 1.25 microM ) , oligodendrocytes displayed a 3- to 4-fold increase ( p=.0002 ) in Al , neurons demonstrated a much smaller ( p=.06 ) increase , and no increase was seen for astrocytes . When incubated with equimolar Al citrate or Al chloride , no increase in cellular Al was seen in any of the three cell types . Oligodendrocytes , astrocytes and neurons all demonstrated greater 59Fe uptake from Fe-Tf than Fe chloride . This uptake could be inhibited by excess Fe-Tf in oligodendrocytes and neurons , but not astrocytes . A small but significant inhibition of 59Fe uptake from Fe-Tf was seen after addition of Al-Tf to the incubation medium of oligodendrocytes , but not neurons or astrocytes . Oligodendrocytes may be particularly vulnerable to the accumulation of excess intracellular Al , and to interference of Al with Fe uptake . Such effects could contribute to Al-induced neurotoxicity if they result in altered myelin formation or maintenance .

MH_train_1199

MH_train_1199

MH_train_1199

interacts_with DB06616?

multiple_choice

Regulation and therapeutic targeting of peptide-activated receptor guanylyl cyclases . Cyclic GMP is a ubiquitous second messenger that regulates a wide array of physiologic processes such as blood pressure , long bone growth , intestinal fluid secretion , phototransduction and lipolysis . Soluble and single-membrane-spanning enzymes called guanylyl cyclases ( GC ) synthesize cGMP . In humans , the latter group consists of P16066 , P20594 , P25092 , GC-E and P51841 , which are also known as P16066 , P20594 , StaR , Ret1-GC and Ret2-GC , respectively . Membrane GCs are activated by peptide ligands such as atrial natriuretic peptide ( P01160 ) , B-type natriuretic peptide ( DB04899 ) , P23582 ( P09543 ) , guanylin , uroguanylin , heat stable enterotoxin and GC-activating proteins . DB04899 and carperitide are clinically approved peptide-based drugs that activate P16066 . CD-NP is an experimental heart failure drug that primarily activates P20594 but also activates P16066 at high concentrations and is resistant to degradation . Inactivating mutations in P20594 cause acromesomelic dysplasia type Maroteaux dwarfism and chromosomal mutations that increase P09543 concentrations are associated with Marfanoid-like skeletal overgrowth . Pump-based P09543 infusions increase skeletal growth in a mouse model of the most common type of human dwarfism , which supports P09543 / P20594 -based therapies for short stature diseases . DB08890 is a peptide activator of P25092 that stimulates intestinal motility and is in late-stage clinical trials for the treatment of chronic constipation . This review discusses the discovery of cGMP , guanylyl cyclases , the general characteristics and therapeutic applications of P16066 , P20594 and P25092 , and emphasizes the regulation of transmembrane guanylyl cyclases by phosphorylation and DB00171 . Association with the P12931 family tyrosyl kinase P07948 triggers a conformational change in the catalytic region of human DB02527 -specific phosphodiesterase HSPDE4A4B . Consequences for rolipram inhibition . The DB02527 -specific phosphodiesterase ( PDE ) HSPDE 4A4B(pde46) selectively bound SH3 domains of P12931 family tyrosyl kinases . Such an interaction profoundly changed the inhibition of DB05876 activity caused by the DB05876 -selective inhibitor rolipram and mimicked the enhanced rolipram inhibition seen for particulate , compared with cytosolic pde46 expressed in COS7 cells . Particulate pde46 co-localized with P07948 kinase in COS7 cells . The unique N-terminal and LR2 regions of pde46 contained the sites for SH3 binding . Altered rolipram inhibition was triggered by SH3 domain interaction with the LR2 region . Purified P07948 SH3 and human P27815 LR2 could be co-immunoprecipitated , indicating a direct interaction . Protein kinase A-phosphorylated pde46 remained able to bind P07948 SH3. pde46 was found to be associated with P12931 kinase in the cytosol of COS1 cells , leading to aberrant kinetics of rolipram inhibition . It is suggested that pde46 may be associated with P12931 family tyrosyl kinases in intact cells and that the ensuing SH3 domain interaction with the LR2 region of pde46 alters the conformation of the PDE catalytic unit , as detected by altered rolipram inhibition . Interaction between pde46 and P12931 family tyrosyl kinases highlights a potentially novel regulatory system and point of signaling system cross-talk . Lectin-like oxidized P01130 -1 ( P78380 ) acts as a receptor for remnant-like lipoprotein particles ( RLPs ) and mediates RLP-induced migration of vascular smooth muscle cells . OBJECTIVE : Remnant-like lipoprotein particles ( RLPs ) have been implicated in atherogenesis especially by diabetic dyslipidemia ; however , their receptor(s) and effects on vascular smooth muscle cells ( VSMCs ) remain unclear . In this study , we examined if lectin-like oxidized P01130 -1 ( P78380 ) acts as a receptor for RLPs and its biological effects in VSMCs . METHODS AND RESULTS : RLPs were isolated from human plasma by immunoaffinity gel containing anti-apolipoprotein A-I and anti-apolipoprotein B-100 monoclonal antibodies . DiI-labeled RLPs were taken up by CHO- P04264 cells stably expressing P78380 but not by wild-type CHO- P04264 cells . RLPs induced P78380 expression and cell migration in bovine VSMCs ( BVSMCs ) , which were significantly suppressed by transfection with P78380 specific siRNAs . Inhibitors of metalloproteinases , epidermal growth factor ( P01133 ) receptor tyrosine kinase , heparin-binding P01133 -like growth factor ( HB- P01133 ) , p38 mitogen-activated protein kinase ( p38 MAPK ) , MAPK kinase ( Q02750 ) and phosphoinositide 3-kinase ( PI3K ) significantly blocked RLP-induced P78380 expression and cell migration of BVSMCs . CONCLUSIONS : The present study provides direct evidence that P78380 is a novel receptor for RLPs in VSMCs . P78380 -mediated uptake of RLPs may thus play important roles in atherogenesis by inducing P78380 expression and VSMC migration especially in the settings of postprandial hyperlipidemia , diabetes and metabolic syndrome . 1alpha,25(OH)2-vitamin D3 inhibits P14210 synthesis and secretion from MG-63 human osteosarcoma cells . Several mesenchymally derived cells , including osteoblasts , secrete hepatocyte growth factor ( P14210 ) . 1alpha,25(OH)(2)-vitamin D(3) [ 1,25(OH)(2)D(3) ] inhibits proliferation and induces differentiation of MG-63 osteoblastic cells . Here we show that MG-63 cells secrete copious amounts of P14210 and that 1,25(OH)(2)D(3) inhibits P14210 production . MG-63 cells also express P08581 ( c- DB00134 ) mRNA , suggesting an autocrine action of P14210 . Indeed , although exogenous P14210 failed to stimulate cellular proliferation , neutralizing endogenous P14210 with a neutralizing antibody inhibited MG-63 cell proliferation ; moreover , inhibiting P14210 synthesis with 1,25(OH)(2)D(3) followed by addition of P14210 rescued hormone-induced inhibition of proliferation . Nonneutralized cells displayed constitutive phosphorylation of c- DB00134 and the mitogen-activated protein kinases mitogen/extracellular signal-regulated kinase ( MEK ) 1 and extracellular signal-regulated kinase ( Erk ) 1/2 , which were inhibited by anti- P14210 antibody . Constitutive phosphorylation of Erk1/2 was also abolished by 1,25(OH)(2)D(3) . Addition of P14210 to MG-63 cells treated with neutralizing P14210 antibody induced rapid phosphorylation of c- DB00134 , Q02750 , and Erk1/2 . Thus endogenous P14210 induces a constitutively active , autocrine mitogenic loop in MG-63 cells . The known antiproliferative effect of 1,25(OH)(2)D(3) on MG-63 cells can be accounted for by the concomitant 1,25(OH)(2)D(3)-induced inhibition of P14210 production . Neuroprotective profile of novel P12931 kinase inhibitors in rodent models of cerebral ischemia . Src kinase signaling has been implicated in multiple mechanisms of ischemic injury , including vascular endothelial growth factor ( P15692 ) -mediated vascular permeability that leads to vasogenic edema , a major clinical complication in stroke and brain trauma . Here we report the effects of two novel Src kinase inhibitors , 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile ( DB06616 ) and 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[4-(4-methypiperazin-1-yl)but-1-ynyl]-3-quinolinecarbonitrile ( SKS-927 ) , on ischemia-induced brain infarction and short- and long-term neurological deficits . Two well established transient [ transient middle cerebral artery occlusion ( tMCAO ) ] and permanent [ permanent middle cerebral artery occlusion ( pMCAO ) ] focal ischemia models in the rat were used with drug treatments initiated up to 6 h after onset of stroke to mimic the clinical scenario . Brain penetration of Src inhibitors , their effect on blood-brain barrier integrity and P15692 signaling in human endothelial cells were also evaluated . Our results demonstrate that both agents potently block P15692 -mediated signaling in human endothelial cells , penetrate rat brain upon systemic administration , and inhibit postischemic Src activation and vascular leakage . Treatment with DB06616 or SKS-927 ( at the doses of 3-30 mg/kg i.v. ) resulted in a dose-dependent reduction in infarct volume and robust protection from neurological impairments even when the therapy was initiated up to 4- to 6-h after tMCAO . Src blockade after pMCAO resulted in accelerated improvement in recovery from motor , sensory , and reflex deficits during a long-term ( 3 weeks ) testing period poststroke . These data demonstrate that the novel Src kinase inhibitors provide effective treatment against ischemic conditions within a clinically relevant therapeutic window and may constitute a viable therapy for acute stroke . Arthrobacter sp. lipase immobilization for preparation of enantiopure masked beta-amino alcohols . Recent reports on immobilization of lipase from Arthrobacter sp . ( P00519 , MTCC 5125 ; IIIM isolate ) on insoluble polymers have shown altered properties including stability and enantioselectivity . Present work demonstrates a facile method for the preparation of enantiopure beta-amino alcohols by modulation of P00519 enzyme properties via immobilization on insoluble as well as soluble supports using entrapment/covalent binding techniques . Efficacies of immobilized P00519 on insoluble supports prepared from tetraethylorthosilicate/aminopropyltriethoxy silane and soluble supports derived from copolymerization of N-vinyl pyrrolidone-allylglycidyl ether ( P01160 type ) /N-vinyl pyrrolidone-glycidyl methacrylate ( GNP type ) for kinetic resolution of masked beta-amino alcohols have been studied vis-à-vis free P00519 enzyme/wet cell biomass . The immobilized lipase on different insoluble/soluble supports has shown 21-110 mg/g protein binding and 30-700 U/g activity for hydrolyzing tributyrin substrate . The findings have shown a significant enhancement in enantioselectivity ( ee 99 % ) vis-à-vis wet cell biomass providing ee 70-90 % for resolution of beta-amino alcohols . Regulation of cyclic AMP response-element binding-protein ( CREB ) by Gq/11-protein-coupled receptors in human SH-SY5Y neuroblastoma cells . Human SH-SY5Y neuroblastoma cells have been used to investigate mechanisms involved in CREB phosphorylation after activation of two endogenously expressed Gq/11-protein-coupled receptors , the M3 muscarinic acetylcholine ( mACh ) and B2 bradykinin receptors . Stimulation with either methacholine or bradykinin resulted in maximal increases in CREB phosphorylation within 1 min , with either a rapid subsequent decrease ( bradykinin ) to basal levels , or a sustained response ( methacholine ) . Inhibitor studies were performed to assess the involvement of a number of potential kinases in signalling to CREB phosphorylation . Removal of extracellular Ca2+ , inhibition of Ca2+/calmodulin-dependent protein kinase II and down-regulation of protein kinase C ( PKC ) resulted in reduced CREB phosphorylation after both M3 mACh and P30411 activation . In contrast , inhibition of Q02750 /2 by U0126 resulted in significantly reduced CREB phosphorylation levels after B2 bradykinin , but not M3 mACh receptor activation . In addition , we demonstrate that maintained phosphorylation of CREB is necessary for CRE-dependent gene transcription as the M3 mACh , but not the P30411 activates both a recombinant CRE-dependent reporter gene , and the endogenous c-Fos gene . These data highlight the involvement of multiple , overlapping signalling pathways linking these endogenous Gq/11-coupled metabotropic receptors to CREB and emphasize the importance of the duration of signalling pathway activation in converting a CREB phosphorylation event into a significant change in transcriptional activity . 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 . Cross-talk between angiotensin II and interleukin-6-induced signaling through Stat3 transcription factor . In cultured neonatal rat cardiac fibroblasts and CHO- P04264 cells expressing angiotensin II ( Ang II ) type 1 receptors ( AT1 ) ( T3CHO/AT1A cell line ) , Ang II induced a delayed tyrosine phosphorylation of Stat3 ( Signal Transducers and Activators of Transcription ) with maximal activation at 2 h . This was in contrast to the rapid tyrosine phosphorylation ( 15-30 min ) of Stat3 by the cytokine interleukin-6 ( P05231 ) . Using T3CHO/AT1A cells , we tested the hypothesis that the delayed tyrosine phosphorylation of Stat3 by Ang II resulted from the induction of an inhibitory pathway ( 0-30 min ) prior to activation ( 1-2 h ) . In support of this hypothesis , we observed that a short treatment of cells with Ang II transiently inhibited the P05231 -induced Stat3 tyrosine phosphorylation . The inhibitory effect of Ang II could be attenuated by exposing the cells to a specific inhibitor of Q02750 , PD98059 . Such modulatory cross-talk between Ang II and P05231 may have relevance in pathophysiological conditions such as cardiac hypertrophy , and in acute phase and inflammatory responses . Two members of a distinct subfamily of 5-hydroxytryptamine receptors differentially expressed in rat brain . We report two serotonin ( 5-hydroxytryptamine , 5-HT ) receptors , MR22 and REC17 , that belong to the G-protein-associated receptor superfamily . MR22 and REC17 are 371 and 357 amino acids long , respectively , as deduced from nucleotide sequence and share 68 % mutual amino acid identity and 30-35 % identity with known catecholamine and 5-HT receptors . Saturable binding of 125I-labeled DB04829 to transiently expressed MR22 in COS-M6 cells was inhibited by ergotamine > methiothepin > 5-carboxamidotryptamine > 5-HT . For REC17 , the rank of potency was ergotamine > 5-carboxamidotryptamine > methiothepin > methysergide > 5-HT . Both were insensitive to P08908 , P28221 or 5-HT2 serotonergic ligands [ 8-hydroxy-2-(di-n-propylamino)tetralin , sumatriptan , and 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ] . The mRNAs encoding MR22 were detected in the P00915 region of hippocampus , the medial habenula , and raphe nuclei . In contrast , mRNAs encoding REC17 were found throughout the rat central nervous system . We propose that REC17 and MR22 , designated as 5-HT5 alpha and 5-HT5 beta , represent a distinct subfamily of 5-HT receptors . The human area postrema and other nuclei related to the emetic reflex express DB02527 phosphodiesterases 4B and 4D . Phosphodiesterase 4 ( DB05876 ) inhibitors , i.e. rolipram , are being extensively investigated as therapeutic agents in several diseases . Emesis is one of the most common side effects of DB05876 inhibitors . Given the fact that the area postrema is considered the chemoreceptor trigger zone for vomiting , the present study investigates the regional distribution and cellular localization of the four gene transcripts of the DB05876 subfamily ( P27815 , Q07343 , Q08493 and Q08499 ) in human brainstem . In situ hybridization histochemistry was used to locate the mRNA distribution of the four DB05876 subfamilies in the area postrema and related nuclei of human postmortem brainstem . We have found that in the brainstem Q07343 and Q08499 mRNA expression is abundant and distributed not only in neuronal cells , but also in glial cells , and on blood vessels . The hybridization signals for Q07343 and Q08499 mRNAs in the area postrema were stronger than those in any other nuclei in the brainstem . They were also found in vomiting-related nuclei such as the nucleus of the solitary tract and the dorsal vagal motor nucleus . These findings suggest that DB02527 signaling modification in the area postrema could mediate the emetic effects of DB05876 inhibitors in human brainstem . 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 . Characterization of the interaction of ingenol 3-angelate with protein kinase C . DB05013 ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate ( PMA ) in WEHI-231 , Q9BPY8 -92 , and Colo-205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO- P04264 cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation . 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 . Targeted agents for the treatment of advanced renal cell carcinoma . Renal cell carcinoma ( RCC ) is a highly treatment-resistant tumor type ; however , advances in elucidating the molecular pathophysiology underlying RCC has led to the identification of promising targets for therapeutic intervention . In clear-cell RCC , mutations to the von Hippel-Lindau ( P40337 ) gene results in the up regulation of many proteins necessary for tumor growth and survival -- such as vascular endothelial growth factor ( P15692 ) , basic fibroblast growth factor ( P09038 ) and platelet derived growth factor ( PDGF ) , which are involved in tumor-initiated angiogenesis . Q16790 and signaling via the epidermal growth factor receptor ( P00533 ) are involved in tumor cell proliferation and are also up regulated by mutation in the P40337 gene . The intracellular messenger pathways phosphoinositide 3-kinase ( PI3K ) and Raf/MEK/ P29323 act as convergence points for positive growth signaling ; the Raf/MEK/ P29323 pathway is also implicated in apoptosis . Several agents in development target P15692 ( bevacizumab ) , the P15692 receptor ( PTK787 , SU11248 , P15692 -trap , and BAY 43-9006 ) , the PDGF receptor ( SU11248 and BAY 43-9006 ) , or the P01133 receptor ( gefitinib , cetuximab , DB01269 , and erlotinib ) . The intracellular Raf/MEK/ P29323 signaling cascade has been targeted at either the level of Raf ( BAY 43-9006 , ISIS 5132 ) or MEK ( CI-1040 , PD184352 and ARRY-142886 ) , and PI3K signaling is disrupted by CCI-779 . DB05304 targets the Q16790 antigen , and PS-341 disrupts the 26S proteasome mediating the degradation of intracellular proteins . Given that multiple pathways contribute to tumor growth , anti-tumor activity may be increased by agents targeting multiple pathways , or by combining agents to allow horizontal or vertical inhibition of multiple pathways . Targeting autocrine and paracrine P15692 receptor pathways inhibits human lymphoma xenografts in vivo . The role of angiogenesis in lymphoproliferative diseases is not well established . We demonstrate here that human lymphoma cells secrete vascular endothelial growth factor ( P15692 ) and express P15692 receptor 1 ( P17948 ) and P35968 . Proliferation of non-Hodgkin lymphoma ( Q9NZ71 ) cells under serum-free conditions was enhanced by the addition of P15692 and was blocked by P17948 - and P35968 -specific antibodies . To differentiate between P15692 -mediated autocrine and paracrine effects on lymphoma growth , NOD/SCID mice engrafted with human diffuse large B-cell lymphoma ( DLBCL ) were treated with species-specific antibodies against human P17948 ( 6.12 ) , human P35968 ( DB06101 ) , murine P17948 ( MF-1 ) , or murine P35968 ( DC101 ) . Treatment with 6.12 or DC101 ( targeting tumor P17948 and host P35968 ) reduced established DLBCL xenograft growth , whereas treatment with DB06101 or MF-1 ( targeting tumor P17948 and host P17948 ) had no effect . Decreased tumor volumes after 6.12 and DC101 treatment correlated with increased tumor apoptosis and reduced vascularization , respectively , supporting the presence of autocrine P17948 - and paracrine P35968 -mediated pathways in lymphomagenesis . Inhibition of paracrine P15692 interactions ( DC101 ) in these models was equivalent to their inhibition with rituximab . Combining DC101 with therapeutic agents ( rituximab , 6.12 , methotrexate ) consistently improved tumor responses over those of single-agent therapy . These data support the further clinical development of VEGFR-targeted approaches for the therapy of aggressive DLBCL . P55157 inhibitor decreases plasma cholesterol levels in P01130 -deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 ) -inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) -2-cyclopentyl-2-[4-[(2,4-dimethyl-9H-pyrido[2,3-b]indol-9-yl)methyl]phenyl]-N- [ ( 1S ) -2-hydroxy-1-phenylethyl ] ethanamide ( DB04852 ) , a new P55157 inhibitor , to low-density lipoprotein ( LDL ) -receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg/kg for 4 weeks . In the 12 mg/kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E/LDL was not altered by DB04852 , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia . The angiotensin II type 1 receptor antagonist Losartan binds and activates bradykinin B2 receptor signaling . The angiotensin II type 1 receptor ( AT1R ) blocker ( ARB ) Losartan has cardioprotective effects during ischemia-reperfusion injury and inhibits reperfusion arrhythmias -effects that go beyond the benefits of lowering blood pressure . The renin-angiotensin and kallikrein-kinin systems are intricately connected and some of the cardioprotective effects of Losartan are abolished by blocking the bradykinin B2 receptor ( P30411 ) signaling . In this study , we investigated the ability of six clinically available ARBs to specifically bind and activate the P30411 . First , we investigated their ability to activate phosphoinositide ( PI ) hydrolysis in COS-7 cells transiently expressing the P30411 . We found that only Losartan activated the P30411 , working as a partial agonist compared to the endogenous ligand bradykinin . This effect was blocked by the P30411 antagonist DB06196 . A competitive binding analysis revealed that Losartan does not significantly compete with bradykinin and does not change the binding affinity of bradykinin on the P30411 . Furthermore , Losartan but not DB00796 mimicked the ability of bradykinin to increase the recovery of contractile force after metabolic stress in rat atrial tissue strips . In conclusion , Losartan is a partial agonist of the P30411 through direct binding and activation , suggesting that P30411 agonism could partly explain the beneficial effects of Losartan .