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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 . 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 . 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 . Agonists and antagonists of DB00644 and -II reduce metastasis formation by triple-negative human breast cancer cells in vivo . Metastasis to bone is a frequent problem of advanced breast cancer . Particularly breast cancers , which do not express estrogen and progesterone receptors and which have no overexpression/amplification of the P04626 -neu gene , so called triple-negative breast cancers , are considered as very aggressive and possess a bad prognosis . About 60 % of all human breast cancers and about 74 % of triple-negative breast cancers express receptors for gonadotropin-releasing hormone ( DB00644 ) , which might be used as a therapeutic target . Recently , we could show that bone-directed invasion of human breast cancer cells in vitro is time- and dose-dependently reduced by DB00644 analogs . In the present study , we have analyzed whether DB00644 analogs are able to reduce metastases of triple-negative breast cancers in vivo . In addition , we have evaluated the effects of DB00644 analogs on tumor growth . To quantify formation of metastasis by triple-negative MDA-MB-435 and MDA-MB-231 human breast cancers , we used a real-time PCR method based on detection of human-specific alu sequences measuring accurately the amount of human tumor DNA in athymic mouse organs . To analyze tumor growth , the volumes of breast cancer xenotransplants into nude mice were measured . We could demonstrate that DB00644 analogs significantly reduced metastasis formation by triple-negative breast cancer in vivo . In addition , we could show that DB00644 analogs significantly inhibited the growth of breast cancer into nude mice . Side effects were not detectable . In conclusion , DB00644 analogs seem to be suitable drugs for an efficacious therapy for triple-negative , P30968 -positive human breast cancers to prevent metastasis formation . 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 . DB06589 , a novel multitargeted kinase inhibitor , shows potent in vitro antitumor activity in gastric cancer cell lines with P21802 amplification . DB06589 is an orally bioavailable , DB00171 -competitive , multitargeted tyrosine kinase inhibitor mainly targeting P35968 and P09619 tyrosine kinases , but the biologic sequences of pazopanib activities beyond antiangiogenesis are poorly defined . We used a panel of 38 gastric cancer cell lines to test the efficacy of pazopanib . In a growth inhibition assay , genomic changes indicated that pazopanib had differential effects on cell growth . Treatment of the KATO-III , OCUM-2M , SNU-16 , and P19526 -39 gastric cancer cell lines harboring P21802 amplification with pazopanib resulted in marked decreases of cell survival with IC50 in ranges of 0.1 to 2.0 μmol/L , whereas the same treatment of those cell lines without P21802 amplification had no growth-inhibitory effects . In the ectopic P21802 -expressing model , treatment with the indicated concentrations of pazopanib significantly inhibited cell growth and colony formation by P21802 -expressing NIH 3T3 cells with wild-type ( WT ) P21802 and mutant P21802 ( S252W ) . DB06589 also selectively suppressed constitutive P21802 signaling and phosphorylation of downstream effectors . In cell-cycle analysis , P21802 -amplified cells underwent cell-cycle arrest at the P55008 -S phase after pazopanib treatment , whereas there were no significant effects on cell-cycle progression in cells without P21802 amplification treated with pazopanib . In addition , pazopanib increased a substantial fraction of sub- P55008 only in P21802 -amplified cells . These findings show that the activation of P21802 signaling by amplification may be a critical mediator of cell proliferation in a small subset of gastric cancer patients and that pazopanib may provide genotype-correlated clinical benefits beyond the setting of highly vascular tumors . P03372 1 ( P03372 ; ERα ) , not Q92731 ( ERβ ) , modulates estrogen-induced sex reversal in the American alligator , a species with temperature-dependent sex determination . All crocodilians and many turtles exhibit temperature-dependent sex determination where the temperature of the incubated egg , during a thermo-sensitive period ( P07996 ) , determines the sex of the offspring . DB00286 play a critical role in sex determination in crocodilians and turtles , as it likely does in most nonmammalian vertebrates . Indeed , administration of estrogens during the P07996 induces male to female sex reversal at a male-producing temperature ( MPT ) . However , it is not clear how estrogens override the influence of temperature during sex determination in these species . Most vertebrates have 2 forms of nuclear estrogen receptor ( P03372 ) : P03372 ( ERα ) and Q92731 ( ERβ ) . However , there is no direct evidence concerning which P03372 is involved in sex determination , because a specific agonist or antagonist for each P03372 has not been tested in nonmammalian species . We identified specific pharmaceutical agonists for each P03372 using an in vitro transactivation assay employing American alligator P03372 and Q92731 ; these were 4,4',4''-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol ( PPT ) and 7-bromo-2-(4-hydroxyphenyl)-1,3-benzoxazol-5-ol ( WAY 200070 ) , respectively . Alligator eggs were exposed to PPT or WAY 200070 at a MPT just before the P07996 , and their sex was examined at the last stage of embryonic development . Estradiol-17β and PPT , but not WAY 200070 , induced sex reversal at a MPT . PPT-exposed embryos exposed to the highest dose ( 5.0 μg/g egg weight ) exhibited enlargement and advanced differentiation of the Müllerian duct . These results indicate that P03372 is likely the principal P03372 involved in sex reversal as well as embryonic Müllerian duct survival and growth in American alligators . SuperTAG methylation-specific digital karyotyping reveals uremia-induced epigenetic dysregulation of atherosclerosis-related genes . BACKGROUND : Accelerated atherosclerosis is a hallmark of chronic kidney disease ( CKD ) . Although the role of epigenetic dysregulation in atherosclerosis is increasingly appreciated , only a few studies focused on epigenetics in CKD-associated cardiovascular disease , virtually all of which assessed epigenetic dysregulation globally . We hypothesized that gene-specific epigenetic dysregulation in CKD exists , affecting genes pertinent to inflammation and atherosclerosis . METHODS AND RESULTS : Ten clinically stable patients undergoing hemodialysis therapy and 10 healthy age- and sex-matched controls were recruited . Genome-wide analysis of DNA methylation was performed by SuperTAG methylation-specific digital karyotyping , in order to identify genes differentially methylated in CKD . Analysis of 27 043 436 tags revealed 4288 genomic loci with differential DNA methylation ( P < 10(-10) ) between hemodialysis patients and control subjects . Annotation of UniTags to promoter databases allowed us to identify 52 candidate genes associated with cardiovascular disease and 97 candidate genes associated with immune/infection diseases . These candidate genes could be classified to distinct proatherogenic processes , including lipid metabolism and transport ( eg , P04035 , P36956 , O75197 , P34913 , and P14324 ) , cell proliferation and cell-cycle regulation ( eg , MIK67 , P04637 , and P18054 ) , angiogenesis ( eg , O15123 , Q9H324 , and P35916 ) , and inflammation ( eg , P50591 , Q9Y6Y9 , P15260 , P0DMV8 , and P42701 ) . CONCLUSIONS : We provide a comprehensive analysis of genome-wide epigenetic alterations in CKD , identifying candidate genes associated with proatherogenic and inflammatory processes . These results may spur further research in the field of epigenetics in kidney disease and point to new therapeutic strategies in CKD-associated atherosclerotic disease . Genomic instability and poor prognosis associated with abnormal P04637 in breast carcinomas . Molecular and immunohistochemical analysis . Alterations of the P04637 gene were analyzed in samples from 87 primary breast cancer patients , using molecular and immunohistochemical approaches . Mutations were detected in 17 % of the samples , using polymerase chain reaction ( PCR ) and constant denaturant gel electrophoresis ( CDGE ) on exons 5-8 of the P04637 gene , and were confirmed by sequencing . Abnormal P04637 protein staining was found in 55 % of the primary samples , using the monoclonal P04637 antibody DO7 . A statistically significant association was found between P04637 mutations and abnormal protein staining ( p = 0.002 ) . Our results suggest that dysfunction of the P04637 protein is associated with tumor progression , as we found an association between P04637 abnormalities and accumulation of genetic lesions , measured as overall allelic imbalance ( AI ) , homogeneously staining regions ( HSR ) and strong P04626 overexpression . Furthermore , patients with P04637 mutation had a highly elevated risk of dying from breast cancer during the study period ( p < 0.001 , RR = 10.68 ) at a median follow-up time of 42 months . Abnormal P04637 staining was much more frequent than the mutations , but it was not of prognostic significance , whereas strong staining was an independent prognostic factor . We therefore conclude that loss of functional P04637 leads to genetic instability , resulting in poorer short-term prognosis , and that only strong staining of P04637 , and not abnormal protein staining in general , is of prognostic significance . 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 . Circulating apoptotic proteins are increased in long-term disease-free breast cancer survivors . Circulating apoptotic proteins are increased in patients with heart failure . We evaluated whether circulating soluble ( s ) apoptosis-related proteins and inflammation markers are increased in long-term disease free breast cancer survivors and associated with cardiotoxicity , and if subgroups could be identified based on the applied treatments . Circulating tumour necrosis factor ( P01375 ) alpha , sTNF-receptor ( sTNF-R ) 1 and 2 , sFas , sFas ligand , sTNF-related apoptosis inducing ligand ( sTRAIL ) and serum P04626 were measured with immunoassay . High-sensitivity P02741 ( HS-CRP ) , fibrinogen , plasma B-type and N-terminal atrial natriuretic peptide ( NT- P01160 and DB04899 ) were also determined . Thirty-four patients with median 6.0 years follow-up and 12 healthy age-matched women were enrolled . Chemotherapy , consisting of five cycles fluorouracil , epirubicin ( 90 mg/m(2) ) , cyclophosphamide ( FEC ) ( n=14 ) or four cycles FEC followed by myeloablation with high-dose carboplatin , cyclophosphamide , thiotepa ( n=20 ) , preceded irradiation and tamoxifen . Circulating apoptosis markers were higher in patients than in controls . No associations with cardiac dysfunction were observed . sFas ligand and sTRAIL were higher in the high-dose than in the standard-dose group . In conclusion , we observed increased circulating apoptotic protein levels in long-term disease-free breast cancer survivors , treated with adjuvant chemoradiotherapy , particularly after myeloablative chemotherapy . The potential relation with late cardiotoxicity of antineoplastic therapy deserves further study . Cl- DB05511 enhances P01375 -α release in peritoneal macrophages stimulated with LPS . P0DMS8 ( A3R ) belongs to the Gi/Gq-coupled receptor family , that leads to the intracellular DB02527 reduction and intracellular calcium increase , respectively . A3R is widely expressed and it can play a crucial role in many patho-physiological conditions , including inflammation . Here we investigate the effect of Cl- DB05511 , A3R agonist , on the production of P01375 -α . We found that Cl- DB05511 enhances LPS-induced P01375 -α release in peritoneal macrophages . This effect is reduced by MRS1191 , A3R antagonist and by forskolin , activator of adenylyl cyclase . pIκBα increased in LPS+Cl- DB05511 -treated macrophages , while total IκB kinase-β ( IKKβ ) reduced . Indeed , p65NF-κB nuclear translocation increased in cells treated with LPS+Cl- DB05511 . Moreover , IMD 0354 , IKKβ inhibitor , significantly abrogated the effect of Cl- DB05511 on P01375 -α release . Inhibition of protein kinase C ( PKC ) significantly reduced Cl- DB05511 -induced P01375 -α release in LPS-stimulated macrophages . Furthermore , LY-294002 , PI3K inhibitor , reduced the P01375 -α production enhanced by Cl- DB05511 , although the phosphorylation status of Akt did not change in cells treated with LPS+Cl- DB05511 than LPS alone . In summary , these data show that Cl- DB05511 is able to enhance P01375 -α production in LPS-treated macrophages in an NF-κB- dependent manner . Expression of cellular retinol-binding protein and lecithin-retinol acyltransferase in developing rat testis . DB00162 deficiency in mammals results in the loss of germ cells on the adluminal side of the blood-testis barrier , suggesting a need for vitamin A that would be supplied by the surrounding Sertoli cells . P09455 ( P09455 ) and lecithin-retinol acyltransferase ( O95237 ) are two proteins found in Sertoli cells that are known to be involved in vitamin A trafficking . To clarify the role of these two proteins in the delivery of vitamin A to developing germ cells , we have examined changes in their cell-specific expression during the onset of puberty in the rat . In adult rats , Sertoli cell expression of P09455 varies with the cycle of the seminiferous epithelium . Here , we demonstrate that differences in the intensity of P09455 immunoreactivity are detectable in Sertoli cells of different tubules as early as postnatal Day 4 , prior to the onset of meiosis . This indicates that variable expression of P09455 by Sertoli cells is established independently of late germ cells and may anticipate the cyclical variation seen in the adult . We further demonstrate that the specific activity of O95237 in rat testis increases tenfold between postnatal Days 20 and 35 . This increase is attributable to the appearance of post-meiotic germ cells : the O95237 activity of microsomes prepared from a round spermatid-enriched cell fraction from post-pubertal rat testis could account for the majority of the O95237 activity observed in the whole testis . The presence of O95237 activity within adluminal germ cells suggests that they receive vitamin A as retinol and synthesize the retinyl esters that have been shown to be present in mature sperm . Rapid purification of a new humanized single-chain Fv antibody/human interleukin-2 fusion protein reactive against P04626 receptor . Human embryonic kidney 293 cells were transfected with plasmid pcDNA-H520C9scFv-rhIL-2 containing a chimeric cDNA encoding the humanized 520C9 scFv/recombinant human P60568 fusion protein ( H520C9scFv-rhIL-2 ) . The transfected cells in plateau growing phase were cultured in serum-free medium for three days . The supernatant was collected , concentrated and purified using an affinity column packed with CNBr-activated Sepharose 4B coupled with anti-rhIL-2 mouse monoclonal antibody . The purified fusion protein was analyzed by ELISA , SDS-PAGE and Western blot . The fusion protein showed only one band in both silver stained electrophoresis gel and Western blot developed by ECL chemiluminescence system . Its molecular weight was confirmed to be about 45 kD . This fusion protein possessed binding specificity against p185 positive SKOV3 and B16/neu cells , and it might stimulate P60568 -dependent CTLL-2 cell proliferation as well . 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 . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . 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 . 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 % . Tra2betal regulates P19 neuronal differentiation and the splicing of FGF-2R and P42262 minigenes . The present study demonstrates that the expression of Tra2beta1 ( Transformer 2-beta1 ) proteins , an SR ( serine/arginine rich ) protein , is developmentally up-regulated in a neural-specific pattern . The up-regulation is also observed in RA ( retinoic acid ) induced neural differentiation of P19 cells . Tra2betal proteins are located in the nuclei of P19 cells , which are consistent with its functional site as an SR protein . The over-expression of Tra2betal proteins promotes RA induced neuronal differentiation of P19 cells . In P19 cells , the splicing of FGF-2R ( fibroblast growth factor receptor 2 ) minigene produces the P21802 form , while the alternative splicing of P42262 ( glutamate receptor subunit B ) minigene generates two products , the Flop and the Truncated isoforms . Tra2betal inhibits the P21802 splicing , but it promotes the Flop splicing . The results therefore suggest that Tra2betal is involved in the regulation of alternative splicing processes during neural development , peculiarly the splicing of FGF-2R and P42262 genes . Both FGF-2R and P42262 genes are known to play important roles in neural differentiation . [ Expression and function identification of extracellular domain 3 of human vascular endothelial growth factor receptor P35968 ] . Vascular Endothelial Growth Factor Receptor-2( P35968 ) plays an important role in stimulating the proliferation of endothelial cells and improving the permeability of blood vessel . We prepared recombinant extracellular domain 3 ( KDR3 ) of human vascular endothelial growth factor receptor-2 in Escherichia coli and studied its specific binding activity with its ligand . The target DNA was synthesized by overlapping PCR , ligated with expression vector p-ET32a and transformed into E. coli Rosetta ( DE3 ) . The soluble fusion protein P10599 -KDR3 was expressed in cytoplasm , which was up to 20 % of total soluble protein in cytoplasm after having been induced by 1 mmol/L DB01862 for 5 h at 30 degrees C . It was characterized to be target protein by Western blotting . The product was purified by CM cation exchange resin and immobilized metal affinity chromatography(IMAC) . Its P15692 -binding activity was determined by ELISA assay and its influence on the propagation of HUVEC induced by P15692 . The protein product showed high ligand binding activity in the ELISA and HUVEC propagation study compared to the control . Therefore , ligand binding active , soluble recombinant extracellular domain 3 of P35968 P35968 was successfully expressed and purified in E. coli , which would be applied to anti-angiogenesis anti-tumor therapy and anti- P35968 antibody development . P04626 signaling modulates the equilibrium between pro- and antiangiogenic factors via distinct pathways : implications for P04626 -targeted antibody therapy . We determined the impact of P04626 signaling on two proangiogenic factors , vascular endothelial growth factor ( P15692 ) and interleukin-8 ( P10145 ) , and on an antiangiogenic factor , thrombospondin-1 ( P07996 -1 ) . Re-expression of P04626 in MCF-7 and T-47D breast cancer cells that endogenously express low levels of P04626 resulted in elevated expression of P15692 and P10145 and decreased expression of P07996 -1 . Inhibition of P04626 with a humanized anti- P04626 antibody ( trastuzumab , or Herceptin ) or a retrovirus-mediated small interfering RNA against P04626 ( siHER2 ) decreased P15692 and P10145 expression , but increased P07996 -1 expression in BT474 breast cancer cells that express high levels of P04626 . These in vitro results were further evaluated by treatment of BT474 xenografts in immunosuppressed mice with trastuzumab . DB00072 inhibited growth of BT474 xenografts and decreased microvascular density associated with downregulation of P15692 and P10145 and with upregulation of P07996 -1 expression . Inhibiting the PI3K-AKT pathway decreased P15692 and P10145 expression . P31749 overexpession increased P15692 and P10145 expression , but did not increase P07996 -1 expression . A p38 kinase inhibitor , SB203580 , instead blocked P07996 -1 expression and a p38 activator , P52564 , increased P07996 -1 expression . DB00072 stimulated sustained p38 activation and SB203580 attenuated the P07996 -1 upregulation induced by trastuzumab . P04626 signaling therefore influences the equilibrium between pro- and antiangiogenic factors via distinct signaling pathways . DB00072 inhibits angiogenesis and tumor growth , at least in part , through activation of the P04626 -p38- P07996 -1 pathway and inhibition of the P04626 -PI3K-AKT- P15692 / P10145 pathway . Neurokinin type-1 receptor antagonist inhibits enhancement of T cell functions by DB05875 in normal and neuromanipulated capsaicin-treated rats . Substance P ( SP ) plays a major role in the regulation of the interaction between immune and nervous systems . SP administration stimulates Con A-induced proliferation of spleen and peripheral blood lymphocytes from normal and neonatally capsaicin treated rats , which correlated with enhanced P60568 production and expression of activation antigens such as P60568 receptor alpha chain ( CD25 ) and RT1B MHC class II molecule . Moreover , SP markedly increased the percentage of P06127 + and P01730 + T lymphocytes in the peripheral blood of capsaicin-treated rats . Concomitant administration of SP with the non-peptide Neurokinin-1 receptor ( P25103 ) antagonist SR140333 completely inhibited the SP-mediated augmentation of Con A-induced PBL proliferation and P60568 production as well as of P01730 + CD25+ and P01730 + RT1B+ T cell numbers in normal and capsaicin-treated rats . DB05790 also blocked the increased percentage of peripheral blood P01730 + T cells induced by SP in capsaicin-treated rats .

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interacts_with DB09068?

multiple_choice

DB02709 and curcumin enhance pancreatic β-cell function by inhibiting phosphodiesterase activity . DB02709 ( RES ) and curcumin ( CUR ) are polyphenols that are found in fruits and turmeric , and possess medicinal properties that are beneficial in various diseases , such as heart disease , cancer , and type 2 diabetes mellitus ( T2DM ) . Results from recent studies have indicated that their therapeutic properties can be attributed to their anti-inflammatory effects . Owing to reports stating that they protect against β-cell dysfunction , we studied their mechanism(s) of action in β-cells . In T2DM , DB02527 plays a critical role in glucose- and incretin-stimulated insulin secretion as well as overall pancreatic β-cell health . A potential therapeutic target in the management of T2DM lies in regulating the activity of phosphodiesterases ( PDEs ) , which degrade DB02527 . Both RES and CUR have been reported to act as PDE inhibitors in various cell types , but it remains unknown if they do so in pancreatic β-cells . In our current study , we found that both RES ( 0.1-10 μmol/l ) and CUR (1-100 pmol/l)-regulated insulin secretion under glucose-stimulated conditions . Additionally , treating β-cell lines and human islets with these polyphenols led to increased intracellular DB02527 levels in a manner similar to DB07954 , a classic PDE inhibitor . When we investigated the effects of RES and CUR on PDEs , we found that treatment significantly downregulated the mRNA expression of most of the 11 PDE isozymes , including Q13370 , O60658 , and Q9Y233 , which have been linked previously to regulation of insulin secretion in islets . Furthermore , RES and CUR inhibited PDE activity in a dose-dependent manner in β-cell lines and human islets . Collectively , we demonstrate a novel role for natural-occurring polyphenols as PDE inhibitors that enhance pancreatic β-cell function . Dysfunction of the P19957 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia . Patients with myeloproliferative disorders ( MPDs ) , such as essential thrombocythemia ( ET ) have increased risk of thrombosis and bleeding , which are major sources of morbidity and mortality . Most P53602 patients have a gain of function mutation in O60674 ( JAK2V617F ) , but little is known how JAK2V617F affects platelet function . Here , we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin ( which couples to O60674 ) on this pathway . In contrast , SFLLRN-mediated P16109 expression , DB00171 secretion , phosphorylation of the PKC substrate pleckstrin , and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets . In addition , thrombopoietin-mediated O60674 phosphorylation was unchanged , suggesting that signaling pathways activated downstream of O60674 are impaired . Indeed , we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the P19957 kinase substrate Akt , and have reduced activation of Rap1 in response to thrombopoietin , DB01277 ,ADP , SFLLRN , and thrombin . This effect was independent of Giα Q9H244 purinergic receptor function as ADP-mediated inhibition of P50552 phosphorylation was unchanged . These results demonstrate that the P19957 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients , resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation . [ 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 . 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 . cDNA microarray reveals signaling pathways involved in hormones expression of human pituitary . Pituitary , a master gland of neuroendocrine system , secretes hormones that orchestrate many physiological processes , under the regulation of multiple signaling pathways . To investigate the genes involved in hormones expression of human pituitary , homemade cDNA microarray containing 14,800 human genes/ESTs were used to profile the gene expression in both fetal and adult pituitaries . Seven hundred and twelve known genes changed over 2-fold between the both tissues . Of which , 23 genes were changed with hormones expression in aging were confirmed by RT-PCR , not only the known regulators such as Pit1 , P43694 , P11474 , GABA-A , and EMK , but also LOC55884 , P51452 , Q9H307 , and O43598 , which had not been reported to be involved in the hormones expression . Correspondingly , the mRNAs of GH , PRL , P01189 , P01222 , DB00094 -beta , and LH-beta , was increased as much as 6- to 20-fold in adult pituitary than those in fetal pituitary , by real-time quantitative RT-PCR assay . In addition , the mRNAs of signaling pathways , such as DB02527 -PKA-CREB , PI3K-Akt , and PKA- P29323 were further investigated . Of them , it was only DB02527 -PKA-CREB pathway , but not PI3K-Akt and PKA- P29323 have the same expressing pattern as hormones . It suggested that cDNA microarray is highly advantages to profile the differential expressed genes that were involved in hormones expression of human pituitary , but it might ignore some responding proteins regulated posttranscriptionally . Proopiomelanocortin but not vasopressin or renin-angiotensin system induces resuscitative effects of central P08908 activation in haemorrhagic shock in rats . The aim of this study was to determine the effectory mechanisms : vasopressin , renin-angiotensin system and proopiomelanocortin-derived peptides ( P01189 ) , partaking in the effects of serotonin through central serotonin 1A receptor ( P08908 ) receptors in haemorrhagic shock in rats . The study was conducted on male Wistar rats . All experimental procedures were carried out under full anaesthesia . The principal experiment included a 2 hour observation period in haemorrhagic shock . Drugs used - a selective P08908 agonist 8-OH-DPAT ( 5 μg/5 μl ) ; V1a receptor antagonist [ β-mercapto-β , β-cyclo-pentamethylenepropionyl(1),O-me- DB00135 (2), DB00125 (8) ] AVP ( 10 μg/kg ) ; angiotensin type I receptor antagonist ( AT1 ) ZD7155 ( 0.5 mg/kg , i.v. ) ; angiotensin-converting-enzyme inhibitor captopril ( 30 mg/kg , i.v. ) ; melanocortin type 4 ( MC4 ) receptor antagonist HS014 ( 5 μg , i.c.v. ) . There was no influence of ZD715 , captopril or blocking of the V1a receptors on changes in the heart rate ( HR ) , mean arterial pressure ( Q96HU1 ) , peripheral blood flow or resistance caused by the central stimulation of P08908 receptors ( P≥0.05 ) . However , selective blocking of central MC4 receptors caused a slight , but significant decrease in HR and Q96HU1 ( P < 0.05 ) . P01189 derivatives acting via the central MC4 receptor participate in the resuscitative effects of 8-OH-DPAT . The angiotensin and vasopressin systems do not participate in these actions . Inhibition of Q13370 augments DB05876 inhibitor-induced apoptosis in a subset of patients with chronic lymphocytic leukemia . PURPOSE : DB02527 phosphodiesterase ( PDE ) 4 is a family of enzymes the inhibition of which induces chronic lymphocytic leukemia ( CLL ) apoptosis . However , leukemic cells from a subset of CLL patients are relatively resistant to treatment with the DB05876 inhibitor rolipram , particularly when this drug is used in the absence of an adenylate cyclase stimulus such as forskolin . Elevated DB02527 levels induce compensatory up-regulation of several cyclic nucleotide PDE families in other model systems . We here examine the hypothesis that CLL cells that survive treatment with rolipram do so as a result of residual PDE activity that is not inhibited by this drug . EXPERIMENTAL DESIGN : We examined by Western analysis the effect of rolipram treatment on CLL expression of Q13370 , P27815 , Q07343 , Q08499 , and Q13946 . We also examined the ability of rolipram ( DB05876 inhibitor ) or cilostamide ( PDE3 inhibitor ) , alone or together , to induce apoptosis or elevate cyclic AMP in leukemic cells from patients with CLL . RESULTS : DB01954 increased levels of Q07343 and , to a variable extent , Q08499 . When combined with forskolin , rolipram also increased levels of a second family of PDEs , Q13370 . Addition of the specific PDE3 inhibitor , cilostamide , modestly augmented rolipram-induced apoptosis in five of seven " rolipram-resistant " CLL samples . CONCLUSIONS : Although this work confirms that DB05876 appears to be the most important PDE target for induction of apoptosis in CLL , combination therapy with PDE3 and DB05876 inhibitors or use of dual-selective drugs may be of benefit in a subset of relatively DB05876 -inhibitor resistant CLL patients . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . Potential advantages of DNA methyltransferase 1 ( P26358 ) -targeted inhibition for cancer therapy . The deoxyribonucleic acid ( DNA ) methyltransferase ( P26358 ) inhibitor DB01262 ( 5-aza-dC ) has been used as a drug in a part of cancer therapy . However , because of its incorporation into DNA during DNA synthesis , 5-aza-dC can cause DNA damage , mutagenesis , and cytotoxicity . In view of the adverse effects of 5-aza-dC , P26358 -targeted inhibition may be a more effective approach than treatment with 5-aza-dC . To address the possibility of P26358 -targeted cancer therapy , we compared the effects of treatment with small interfering ribonucleic acids ( siRNAs ) specific for P26358 or DNMT3b and treatment with 5-aza-dC on transcription , cell growth , and DNA damage in gastric cancer cells . We found that P26358 -targeted inhibition induced the re-expression and reversed DNA methylation of five ( CDKN2A , RASSF1A , P32314 , Q13761 , and AKAP12B ) out of seven genes examined , and 5-aza-dC reactivated and demethylated all seven genes . In contrast , DNMT3b siRNAs did not show any effect . Furthermore , the double knockdown of P26358 and DNMT3b did not show a synergistic effect on gene re-expression and demethylation . In addition , P26358 siRNAs showed an inhibitory effect of cell proliferation in the cancer cells and the induction of cell death without evidence of DNA damage , whereas treatment with 5-aza-dC caused DNA damage as demonstrated by the comet assay . These results provide a rationale for the development of a P26358 -targeted strategy as an effective epigenetic cancer therapy . Molecular cloning of Q9GZP0 , a novel growth factor homologous to Q9NRA1 / Q9NRA1 /fallotein . Q9NRA1 ( Q9NRA1 ) /platelet-derived growth factor ( PDGF ) -C/fallotein has a unique two-domain structure , as it contains two regions homologousto CUB and PDGF/vascular endothelial growth factor ( P15692 ) domains . In this study , we isolateda novel gene homologous to Q9NRA1 / Q9NRA1 /fallotein , and named Q9GZP0 . The culture supernatant of CHO- P04264 cells stably transfected with Q9GZP0 showed mitogenic activity as Q9NRA1 / Q9NRA1 /fallotein did . Although Q9GZP0 and Q9NRA1 / Q9NRA1 /fallotein might be the members of the PDGF/ P15692 superfamily of growth factors , they were categorized into a new subfamily in addition to PDGF and P15692 subfamilies . 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 ) . 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 . 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 . 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 . 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 . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . 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 . Characterization of DB02527 degradation by phosphodiesterases in the accessory olfactory system . To characterize the potential role of DB02527 in pheromone transduction , we have examined the occurrence of cyclic nucleotide phosphodiesterases ( PDEs ) in the mouse vomeronasal organ ( VNO ) . We show that the DB02527 -specific isoforms P27815 and Q08499 are found preferentially in the apical and basal layers , respectively , of the VNO neuroepithelium and in the rostral ( P27815 ) and caudal ( Q08499 ) portions of the accessory olfactory bulb glomerular layer . Assays for DB02527 hydrolysis showed that PDE activity in VNO homogenates was about half that measured in the cerebral cortex and olfactory epithelium , and the proportion of total activity inhibited by rolipram , a DB05876 -specific inhibitor , was approximately 40 % . Activity in the VNO was enhanced 60 % by Ca(2+) and calmodulin ( P62158 ) , implicating the presence of Ca(2+)/ P62158 -dependent PDE1 . Zaprinast , which is known to inhibit Q14123 isoforms , completely suppressed Ca(2+)/ P62158 -stimulated activity and , together , zaprinast and rolipram inhibited DB02527 hydrolysis by approximately 70 % . Our results suggest that PDE1 and DB05876 isoforms are the primary source of DB02527 degradation in the VNO . P05231 enriched lung cancer stem-like cell population by inhibition of cell cycle regulators via P26358 upregulation . Tumors are influenced by a microenvironment rich in inflammatory cytokines , growth factors and chemokines , which may promote tumor growth . P05231 ( P05231 ) is a multifunctional cytokine and known as a regulator of immune and inflammation responses . P05231 has also been reported to be associated with tumor progression and chemoresistance in different types of cancers . In our study , we demonstrated that P05231 enriches the properties of lung cancer stem-like cells in A549 lung cancer cells cultured in spheroid medium . P05231 also promotes sphere formation and stem-like properties of A549 cells by enhancing cell proliferation . Methylation-specific polymerase chain reaction ( PCR ) was performed and revealed that P05231 increased methylation of p53 and P38936 in A549 cancer cells . Western blot analysis and quantitative real-time PCR demonstrated that P05231 increased the expression of DNA methyltransferase 1 ( P26358 ) in A549 cells cultured in spheroid medium , but not the expression of DNMT3a or DNMT3b . Knockdown of P26358 eliminated P05231 -mediated hypermethylation of cell cycle regulators and enrichment of lung cancer stem-like properties . In conclusion , our study , for the first time , shows that the P05231 / O60674 / P40763 pathway upregulates P26358 and enhances cancer initiation and lung cancer stem cell ( CSC ) proliferation by downregulation of p53 and P38936 resulting from DNA hypermethylation . Upon blockage of the P05231 / O60674 / P40763 pathway and inhibition of P26358 , the proliferation of lung CSCs was reduced and their formation of spheres and ability to initiate tumor growth were decreased . These data suggest that targeting of the P05231 / O60674 / P40763 signaling pathway and P26358 may become important strategies for treating lung cancer . 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 . 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 .

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MH_train_1502

MH_train_1502

interacts_with DB01016?

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 . Lycopene and other carotenoids inhibit estrogenic activity of 17beta-estradiol and genistein in cancer cells . Epidemiological evidence suggests that carotenoids prevent several types of cancer , including mammary and endometrial cancers . On the other hand , such studies have also shown that estrogens are the most important risk factors for these cancer types . DB01645 , the phytoestrogen mainly found in soy , also shows significant estrogenic activity when tested at concentrations found in human blood . The aim of this study was to determine whether carotenoids inhibit signaling of steroidal estrogen and phytoestrogen which could explain their cancer preventive activity . Similar to the known effect of 17beta-estradiol ( E(2) ) , treatment of breast ( T47D and MCF-7 ) and endometrial ( ECC-1 ) cancer cells with phytoestrogens induced cell proliferation , cell-cycle progression and transactivation of the estrogen response element ( ERE ) . However , each of the tested carotenoids ( lycopene , phytoene , phytofluene , and beta-carotene ) inhibited cancer cell proliferation induced by either E(2) or genistein . The inhibition of cell growth by lycopene was accompanied by slow down of cell-cycle progression from P55008 to S phase . Moreover , the carotenoids inhibited estrogen-induced transactivation of ERE that was mediated by both estrogen receptors ( ERs ) ERalpha and ERbeta . The possibility that this inhibition results from competition of carotenoid-activated transcription systems on a limited pool of shared coactivators with the ERE transcription system was tested . Although cotransfection of breast and endometrial cancer cells with four different coactivators ( Q15788 , P12931 -2 , Q9Y6Q9 , and DRIP ) strongly stimulated ERE reporter gene activity , it did not oppose the inhibitory effect of carotenoids . These results suggest that dietary carotenoids inhibit estrogen signaling of both 17beta-estradiol and genistein , and attenuate their deleterious effect in hormone-dependent malignancies . 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 . Blockage of JNK pathway enhances arsenic trioxide-induced apoptosis in human keratinocytes . Arsenic is well known as a carcinogen predisposing humans to some severe diseases and also as an effective medicine for treating acute promyelocytic leukemia , syphilis , and psoriasis . Multiple active mechanisms , including cell cycle arrest and apoptosis , have been proposed in therapy ; however , the opposing effects of arsenic remain controversial . Our previous study found that arsenic trioxide ( ATO ) -induced activation of P38936 ( P38936 /CIP1) ( P38936 ) led to A431 cell death through the antagonistic effects of the signaling of P27361 /2 and P45983 . In the current study , the inhibitory effects of P45983 on ATO-induced P38936 expression were explored . Over-expression of P45983 in A431 cells could inhibit P38936 expression , which was associated with Q13547 and Q15583 . Using the Q86UG4 pull-down assay and fluorescence resonance energy transfer analysis , N-terminal domain ( amino acids 1-108 ) of Q15583 , critical to its binding with c-Jun , was found . Using reporter assays , requirement of the C-terminal domain ( amino acids 138-272 ) of Q15583 to suppress ATO-induced P38936 expression was observed . Thus , the domains of Q15583 that carried out its inhibitory effects on P38936 were identified . Finally , treatment with JNK inhibitor SP600125 could enhance ATO-induced apoptosis of HaCaT keratinocytes by using flow cytometry . P45984 phosphorylates endothelial nitric oxide synthase at serine 116 and regulates nitric oxide production . The c-Jun N-terminal kinases ( JNKs ) belonging to the mitogen-activated protein kinase ( MAPK ) superfamily play important roles in foam-cell formation , hypercholesterolemia-mediated endothelial dysfunction , and the development of obesity . Although decreased nitric oxide ( NO ) production via decreased phosphorylation of endothelial NO synthase at serine 1179 ( P29474 - DB00133 (1179) ) was reported to be partly involved in P45984 -derived endothelial dysfunction , P45984 seems likely to be indirectly involved in this signaling pathway . Here , using bovine aortic endothelial cells , we examined whether P45984 directly phosphorylated P29474 - DB00133 (116) , a putative substrate site for the MAPK superfamily , and this phosphorylation resulted in decreased NO release . JNK inhibitor SP60012 increased NO release in a time- and dose-dependent manner , which was accompanied by increased P29474 - DB00133 (116) phosphorylation . Purified P45984 directly phosphorylated P29474 - DB00133 (116)in vitro . Ectopic expression of dominant negative P45984 repressed P29474 - DB00133 (116) phosphorylation and increased NO production . Coimmunoprecipitation and confocal microscopy studies revealed a colocalization of P29474 and P45984 . However , all these observed effects were not manifested when P45983 probes were used . Overall , this study indicates that P45984 is a physiological kinase responsible for P29474 - DB00133 (116) phosphorylation and regulates NO production . DB01645 stimulates electrogenic Cl(-) secretion in mouse jejunum . We used the short-circuit current ( I(sc) ) technique to investigate the effects of the isoflavone genistein on the electrogenic Cl(-) secretion of the mouse jejunum . DB01645 stimulated a sustained increase in I(sc) that was dose dependent . DB00887 inhibited 76 +/- 5 % of the genistein-stimulated I(sc) consistent with activation of Cl(-) secretion . DB01645 failed to stimulate I(sc) following maximal activation of the DB02527 pathway by forskolin . In addition , forskolin had a reduced effect on I(sc) of the mouse jejunum in the presence of genistein . DB01016 , a blocker of P13569 , eliminated the genistein-stimulated increase of I(sc) and reduced the forskolin-activated I(sc) . Clotrimazole , a Ca(2+)-activated K(+) channel blocker , failed to reduce the genistein-stimulated I(sc) . Vanadate , a blocker of tyrosine-dependent phosphatases , reduced the genistein-activated I(sc) . Tyrphostin A23 , a tyrosine kinase inhibitor , reduced basal I(sc) , after which genistein failed to stimulate I(sc) . These data suggest that genistein activated a sustained Cl(-) secretory response of the mouse jejunum and that the effect of genistein was via a tyrosine-dependent phosphorylation pathway . Identification of a transcriptionally active peroxisome proliferator-activated receptor alpha -interacting cofactor complex in rat liver and characterization of Q9BYK8 as a coactivator . Q07869 ( Q07869 alpha ) plays a central role in the cell-specific pleiotropic responses induced by structurally diverse synthetic chemicals designated as peroxisome proliferators . Transcriptional regulation by liganded nuclear receptors involves the participation of cofactors that form multiprotein complexes to achieve cell- and gene-specific transcription . Here we report the identification of such a transcriptionally active Q07869 alpha-interacting cofactor ( PRIC ) complex from rat liver nuclear extracts that interacts with full-length Q07869 alpha in the presence of ciprofibrate , a synthetic ligand , and leukotriene B(4) , a natural ligand . The liganded Q07869 alpha-PRIC complex enhanced transcription from a peroxisomal enoyl- DB01992 hydratase/l-3-hydroxyacyl- DB01992 dehydrogenase bifunctional enzyme gene promoter template that contains peroxisome proliferator response elements . Rat liver PRIC complex comprises some 25 polypeptides , and their identities were established by mass spectrometry and limited sequence analysis . Eighteen of these peptides contain one or more LXXLL motifs necessary for interacting with nuclear receptors . PRIC complex includes known coactivators or coactivator-binding proteins ( CBP , Q15788 , PBP , PRIP , PIMT , O75448 , Q09428 -2 , and P20142 -1 ) , other proteins that have not previously been described in association with transcription complexes ( CHD5 , TOG , and Q8WYB5 ) , and a few novel polypeptides designated PRIC300 , -285 , -215 , -177 , and -145 . We describe the cDNA for Q9BYK8 , which contains five LXXLL motifs . It interacts with Q07869 alpha and acts as a coactivator by moderately stimulating Q07869 alpha-mediated transcription in transfected cells . We conclude that liganded Q07869 alpha recruits a distinctive multiprotein complex from rat liver nuclear extracts . The composition of this complex may provide insight into the basis of tissue and species sensitivity to peroxisome proliferators . The thioredoxin reductase inhibitor auranofin triggers apoptosis through a Bax/Bak-dependent process that involves peroxiredoxin 3 oxidation . P30044 ( TrxR ) is a key selenoprotein antioxidant enzyme and a potential target for anti-cancer drugs . One potent inhibitor of TrxR is the gold ( I ) compound auranofin , which can trigger mitochondrial-dependent apoptosis pathways . The exact mechanism of apoptosis induction by auranofin is not yet clear , but there are indications that mitochondrial oxidative stress is a central event . We assessed the redox state of the peroxiredoxins ( Prxs ) in Jurkat T-lymphoma cells treated with auranofin , and found that mitochondrial Prx3 was considerably more sensitive to oxidation than the cytosolic Prx1 and 2 , indicating selective mitochondrial stress . Prx3 oxidation was detected at apoptotic doses of auranofin in several cell types , and occurred before other mitochondrial events including cytochrome c release and mitochondrial depolarisation . DB00995 was also able to sensitise U937 cells to P01375 -mediated apoptosis . DB00995 -induced apoptosis was effectively blocked by the overexpression of Bcl-2 , and Bax/Bak deficient mouse embryonic fibroblasts were also resistant to apoptosis , indicating a central role for the pro-apoptotic proteins of this family in auranofin-triggered apoptosis . DB00995 exposure inhibited the proliferation of apoptosis-resistant cells , and at higher doses of auranofin could cause cell death through necrosis . We conclude that auranofin induces apoptosis in cells through a Bax/Bak-dependent mechanism associated with selective disruption of mitochondrial redox homeostasis in conjunction with oxidation of Prx3 . Oxidation of alcohols and reduction of aldehydes derived from methyl- and dimethylpyrenes by cDNA-expressed human alcohol dehydrogenases . Some methylated pyrenes can form DNA adducts in rat tissues after benzylic hydroxylation and sulpho conjugation . However , oxidation of the intermediate alcohols to carboxylic acids is an important competing pathway leading to detoxification . We previously showed that co-administration of ethanol or DB01213 strongly enhances DNA adduct formation by 1-hydroxymethylpyrene , indicating an involvement of alcohol dehydrogenases ( ADHs ) in the detoxification . This mechanism may be involved in the observed synergism of smoking and alcohol consumption in certain human cancers . In a preceding study , cDNA-expressed human P00325 efficiently oxidised 1- , 2- and 4-hydroxymethylpyrene ; these reactions were inhibited in the presence of ethanol or DB01213 . Here we report that P00326 , P00326 and P08319 also show substantial activity towards these substrates and two further congeners , 1-hydroxymethyl-6-methylpyrene and 1-hydroxymethyl-8-methylpyrene . All four DB00067 forms also catalysed the reverse reaction , implying that the aldehydes have to be sequestered by other enzymes , such as aldehyde dehydrogenases , for final detoxification . P00326 and P08319 activities towards hydroxymethylpyrenes were more strongly inhibited in the presence of ethanol and DB01213 than those of P00325 . P00326 was only inhibited at very high concentrations of the modulators . In conclusions , several human ADHs are capable of detoxifying benzylic alcohols of alkylated polycyclic aromatic hydrocarbons . However , some competing substrates and inhibitors may affect all these redundant detoxification systems , although to various extents . Genetic variants in epidermal growth factor receptor pathway genes and risk of esophageal squamous cell carcinoma and gastric cancer in a Chinese population . The epidermal growth factor receptor ( P00533 ) signaling pathway regulates cell proliferation , differentiation , and survival , and is frequently dysregulated in esophageal and gastric cancers . Few studies have comprehensively examined the association between germline genetic variants in the P00533 pathway and risk of esophageal and gastric cancers . Based on a genome-wide association study in a Han Chinese population , we examined 3443 SNPs in 127 genes in the P00533 pathway for 1942 esophageal squamous cell carcinomas ( ESCCs ) , 1758 gastric cancers ( GCs ) , and 2111 controls . SNP-level analyses were conducted using logistic regression models . We applied the resampling-based adaptive rank truncated product approach to determine the gene- and pathway-level associations . The P00533 pathway was significantly associated with GC risk ( P = 2.16×10(-3) ) . Gene-level analyses found 10 genes to be associated with GC , including P06241 , P45983 , P45985 , P08754 , Q02750 , Q9Y490 , P16471 , P16885 , Q9UBS0 , and Q92569 ( P < 0.05 ) . For ESCC , we did not observe a significant pathway-level association ( P = 0.72 ) , but gene-level analyses suggested associations between P08754 , Q04844 , O96013 , O00401 , and Q96J02 , and ESCC ( P < 0.05 ) . Our data suggest an association between specific genes in the P00533 signaling pathway and risk of GC and ESCC . Further studies are warranted to validate these associations and to investigate underlying mechanisms . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . 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 . 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 . Virtual screening for novel openers of pancreatic K( DB00171 ) channels . Ligand-based virtual screening approaches were applied to search for new chemotype KCOs activating Kir6.2/ Q09428 KATP channels . A total of 65 208 commercially available compounds , extracted from the ZINC archive , served as database for screening . In a first step , pharmacokinetic filtering via VolSurf reduced the initial database to 1913 compounds . Afterward , six molecules were selected as templates for similarity searches : similarity scores , obtained toward these templates , were calculated with the GRIND , P20292 , and TOPP approaches , which differently encode structural information into potential pharmacophores . In this way , we obtained 32 hit candidates , 16 via GRIND and eight each via P20292 and TOPP . For biological testing of the hit candidates , their effects on membrane potentials in P29320 293 cells expressing Kir6.2/ Q09428 were studied . GRIND , P20292 , and TOPP all yielded hits , but no method top-ranked all the actives . Thus , parallel application of different approaches probably improves hit detection . Exposure of P13671 glioma cells to Pb(II) increases the phosphorylation of p38(MAPK) and P45983 /2 but not of P27361 /2 . Pb(II) is a neurotoxic pollutant that produces permanent cognitive deficits in children . Pb(II) can modulate cell signaling pathways and cell viability in a variety of cell types . However , these actions are not well demonstrated on glial cells , which represent an important target for metals into the central nervous system . The present work was undertaken to determine the ability of Pb(II) in modulating the activity of mitogen activated protein kinases ( MAPKs ) in cultures of P13671 rat glioma cells , a useful functional model for the study of astrocytes . Additionally , cell viability was analyzed by measurement of MTT reduction . Cells were exposed to lead acetate 0.1 , 1 , 10 microM for 24 and 48 h . MAPKs activation - in particular P27361 /2 , p38(MAPK) and P45983 /2 - were analyzed by western blotting . Results showed that 10 microM Pb(II) treatment for 24 h caused a discrete stimulation of p38(MAPK) phosphorylation . However , 1 and 10 microM Pb(II) treatment for 48 h provoked a significant stimulation in the phosphorylation state of p38(MAPK) and P45983 /2 . The phosphorylation state of P27361 /2 was not modified by any Pb(II) treatment . Moreover , data indicate that at 48 h treatment even 1 microM Pb(II) can be cytotoxic , causing impairment on cell viability . Therefore , depending on a long incubation period , a significant concomitant activation of p38(MAPK) and P45983 /2 by Pb(II) took place in parallel with the impairment of P13671 glioma cells viability . Certain background factors exhibit an association with an increased risk for pancreatic calcification among Japanese male alcoholics . OBJECTIVE : This was a cross-sectional study conducted from April 2003 through March 2004 to investigate the background factors related to pancreatic calcification ( PC ) in male Japanese alcoholics . METHODS AND RESULTS : Helical computed tomography examination revealed PC in 44 of 263 alcoholics , and this group was further divisible into 3 subgroups : " scant " ( n = 24 ) , " moderate " ( n = 6 ) , and " extensive " PC subgroups ( n = 14 ) . The extensive subgroup was associated with larger daily ethanol consumption ( P = 0.05 ) and high-alcohol beverages , such as whisky ( P = 0.02 ) . The moderate subgroup was associated with a longer duration of habitual drinking ( P = 0.04 ) , whereas the scant PC group was associated with never having smoked ( P = 0.05 ) and with low-alcohol beverages , such as beer ( P = 0.09 ) . None of the 40 subjects with inactive mitochondrial aldehyde dehydrogenase ( P05091 *2 allele ) exhibited PC ( P = 0.004 ) . Heterozygous alcohol dehydrogenase 2 genotype ( P00325 *1/2*2 ) exhibited an association with the scant subgroup ( P = 0.02 ) . The TG12 repeats in the cystic fibrosis transmembrane conductance regulator ( P13569 ) gene tended to have a weak association with PC . CONCLUSION : Drinking behavior , smoking status , and genetic backgrounds are associated with PC and are likely to increase the risk for alcoholic chronic pancreatitis . 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 . 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 . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Intrauterine growth retardation is associated with reduced activity and expression of the cationic amino acid transport systems y+/hCAT-1 and y+/hCAT-2B and lower activity of nitric oxide synthase in human umbilical vein endothelial cells . Intrauterine growth retardation ( IUGR ) is associated with vascular complications leading to hypoxia and abnormal fetal development . The effect of IUGR on L-arginine transport and nitric oxide ( NO ) synthesis was investigated in cultures of human umbilical vein endothelial cells ( HUVECs ) . IUGR was associated with membrane depolarization and reduced L-arginine transport ( V(max)= 5.8+/-0.2 versus 3.3+/-0.1 pmol/microg protein per minute ) , with no significant changes in transport affinity ( K(m)=159+/-15 versus 137+/-14 micromol/L ) . L- DB00125 transport was trans-stimulated ( 8- to 9-fold ) in cells from normal and IUGR pregnancies . IUGR was associated with reduced production of L-[3H]citrulline from L-[3H] arginine , lower nitrite and intracellular L-arginine , L-citrulline , and cGMP . IUGR decreased hCAT-1 and hCAT-2B mRNA , and increased P29474 mRNA and protein levels . IUGR-associated inhibition of L-arginine transport and NO synthesis , and membrane depolarization were reversed by the NO donor S-nitroso-N-acetyl-L, DB00859 . In summary , endothelium from fetuses with IUGR exhibit altered L-arginine transport and NO synthesis ( L-arginine/NO pathway ) , reduced expression and activity of hCAT-1 and hCAT-2B and reduced P29474 activity . Alterations in L-arginine/NO pathway could be critical for the physiological processes involved in the etiology of IUGR in human pregnancies . 5-Lipoxygenase-activating protein ( P20292 ) inhibitors . Part 4 : development of 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid ( AM803 ) , a potent , oral , once daily P20292 inhibitor . The potent P09917 -activating protein ( P20292 ) inhibitor 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid 11cc is described ( AM803 , now GSK2190915 ) . Building upon DB05225 ( 1 ) ( Hutchinson et al. J. Med Chem.2009 , 52 , 5803-5815 ; Stock et al. Bioorg. Med. Chem. Lett. 2010 , 20 , 213-217 ; Stock et al. Bioorg. Med. Chem. Lett.2010 , 20 , 4598-4601 ) , SAR studies centering around the pyridine moiety led to the discovery of compounds that exhibit significantly increased potency in a human whole blood assay measuring Q06643 (4) inhibition with longer drug preincubation times ( 15 min vs 5 h ) . Further studies identified 11cc with a potency of 2.9 nM in P20292 binding , an IC(50) of 76 nM for inhibition of Q06643 (4) in human blood ( 5 h incubation ) and excellent preclinical toxicology and pharmacokinetics in rat and dog . 11cc also demonstrated an extended pharmacodynamic effect in a rodent bronchoalveolar lavage ( BAL ) model . This compound has successfully completed phase 1 clinical studies in healthy volunteers and is currently undergoing phase 2 trials in asthmatic patients . Loss of Androgen-Regulated MicroRNA 1 Activates P12931 and Promotes Prostate Cancer Bone Metastasis . Bone metastasis is the hallmark of progressive and castration-resistant prostate cancers . MicroRNA 1 ( miR-1 ) levels are decreased in clinical samples of primary prostate cancer and further reduced in metastases . P12931 has been implicated as a critical factor in bone metastasis , and here we show that P12931 is a direct target of miR-1 . In prostate cancer patient samples , miR-1 levels are inversely correlated with P12931 expression and a P12931 -dependent gene signature . Ectopic miR-1 expression inhibited extracellular signal-regulated kinase ( P29323 ) signaling and bone metastasis in a xenograft model . In contrast , P12931 overexpression was sufficient to reconstitute bone metastasis and P29323 signaling in cells expressing high levels of miR-1 . P10275 ( AR ) activity , defined by an AR output signature , is low in a portion of castration-resistant prostate cancer . We show that AR binds to the miR-1-2 regulatory region and regulates miR-1 transcription . Patients with low miR-1 levels displayed correlated low canonical AR gene signatures . Our data support the existence of an AR-miR-1- P12931 regulatory network . We propose that loss of miR-1 is one mechanistic link between low canonical AR output and P12931 -promoted metastatic phenotypes .

MH_train_1503

MH_train_1503

MH_train_1503

interacts_with DB01151?

multiple_choice

On the hepatic mechanism of HDL assembly by the O95477 /apoA-I pathway . The mechanism for the assembly of HDL with cellular lipid by O95477 and helical apolipoprotein was investigated in hepatocytes . Both HepG2 cells and mouse primary culture hepatocytes produced HDL with apolipoprotein A-I ( apoA-I ) whether endogenously synthesized or exogenously provided . DB01599 , an O95477 inactivator , inhibited these reactions , as well as the reversible binding of apoA-I to HepG2 . Primary cultured hepatocytes of O95477 -deficient mice also lacked HDL production regardless of the presence of exogenous apoA-I . HepG2 cells secreted apoA-I into the medium even when O95477 was inactivated by probucol , but it was all in a free form as HDL production was inhibited . When a lipid-free apoA-I-specific monoclonal antibody , 725-1E2 , was present in the culture medium , production of HDL was suppressed , whether with endogenous or exogenously added apoA-I , and the antibody did not influence HDL already produced by HepG2 cells . We conclude that the main mechanism for HDL assembly by endogenous apoA-I in HepG2 cells is an autocrine-like reaction in which apoA-I is secreted and then interacts with cellular O95477 to generate HDL . 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 . 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 . Virion attachment and entry : HIV gp120 Env biotinylation , gp120 Env , or integrin ligand-binding assay . The HIV-1 entry receptors are P01730 and a chemokine receptor ( P51681 or P61073 ) . In addition it has recently been demonstrated that HIV-1 gp120 binds to and signals through integrin α4β7 , the gut-homing receptor ( Arthos et al. , Nat Immunol 9(3):301-309 , 2008 ) . Integrin α4β7 is not an entry receptor for HIV-1 , although it can facilitate virion attachment to target cells ( Arthos et al. , Nat Immunol 9(3):301-309 , 2008 ; Cicala et al. , Proc Natl Acad Sci U S A 106:20877-20882 , 2009 ) . Recombinant HIV-1 gp120s bind to integrin α4β7 in a manner similar to its natural ligands ( Q13477 , V- P62158 -1 , fibronectin ) ( Andrew et al. , J Immunol 153:3847-3861 , 1994 ) . gp120-α4β7 interactions are detected in a manner similar to assays developed for the natural ligands of α4β7 . In this chapter we describe a method for the analysis of integrin-gp120 binding via a cell-based binding assay . In vitro ligand-integrin affinity can be modified by the presence of divalent cations ( Mn(2+) , Mg(2+) , Ca(2+) ) ( Leitinger et al. , Leitinger Biochim Biophys Acta 1498:91-98 , 2000 ) . Here we describe a protocol to detect biotinylated recombinant HIV-1 gp120 binding to integrin α4β7 in both primary cells and cell lines expressing the gut-homing receptor . 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 . 2(3H)-benzoxazolone and bioisosters as " privileged scaffold " in the design of pharmacological probes . The 2(3H)-benzoxazolone heterocycle and its bioisosteric surrogates ( such as 2(3H)-benzothiazolinone , benzoxazinone , etc. ) have received considerable attention from the medicinal chemists owing to their capacity to mimic a phenol or a catechol moiety in a metabolically stable template . These heterocycles and pyrocatechol have indeed similar pKa 's , electronic charge distribution , and chemical reactivity . Therapeutic applications of this template are very broad , and range from analgesic anti-inflammatory compounds ( including P37231 antagonists ) to antipsychotic and neuroprotective anticonvulsant compounds . High affinity ligands have been obtained also for dopaminergic ( D2 and D4 ) , serotoninergic ( P08908 and P28223 ) , sigma-1 and sigma-2 receptors . Owing to the high number of positive hits encountered with this heterocycle and its congeners , 2(3H)-benzoxazolone template certainly deserves the title of " privileged scaffold " in medicinal chemistry . Interactions between genetic variants of folate metabolism genes and lifestyle affect plasma homocysteine concentrations in the Boston Puerto Rican population . OBJECTIVE : To investigate genetic and lifestyle factors and their interactions on plasma homocysteine ( Hcy ) concentrations in the Boston Puerto Rican population . DESIGN : Cross-sectional study . Plasma concentrations of Hcy , folate , vitamin B12 and pyridoxal phosphate were measured , and genetic polymorphisms were determined . Data on lifestyle factors were collected in interviews . SETTING : A population survey of health and nutritional measures . SUBJECTS : A total of 994 Puerto Rican men and women residing in the Boston metropolitan area . RESULTS : Smoking status was positively associated with plasma Hcy . Genetic polymorphisms P42898 677C→T , Q04609 1561C→T , Q04609 rs647370 and Q96NT5 928A→G interacted significantly with smoking for Hcy . P42898 1298A→C ( P = 0·040 ) and Q96NT5 928A→G ( P = 0·002 ) displayed significant interactions with alcohol intake in determining plasma Hcy . Subjects with Q96NT5 928GG genotype had significantly higher plasma Hcy concentrations compared with carriers of the A allele ( AA+AG ; P = 0·030 ) among non-drinking subjects . When consuming alcohol , GG subjects had lower plasma Hcy levels compared with AA+AG subjects . Physical activity interacted significantly with Q99707 2756A→G in determining plasma Hcy ( P for interaction = 0·002 ) . Smoking interacted with physical activity for plasma Hcy ( P for interaction = 0·023 ) . CONCLUSIONS : Smoking and drinking were associated plasma Hcy concentrations . Genetic variants involved in folate metabolism further modify the effects of lifestyle on plasma Hcy . Effect of the O00206 antagonist eritoran on retinochoroidal inflammatory damage in a rat model of endotoxin-induced inflammation . PURPOSE : We investigated the effect of eritoran , a O00206 antagonist , on retinochoroidal inflammatory damage in an endotoxin-induced inflammatory rat model . METHODS : Endotoxin-induced inflammatory model was obtained by intraperitoneal injection of 1.5 mg/kg lipopolysaccharide ( LPS ) . Group 1 had control rats ; in groups 2-3 LPS and 0.5 mg/kg sterile saline were injected ; and in groups 4-5 LPS and 0.5 mg/kg eritoran were injected . Blood samples were taken and eyes were enucleated after 12 hours ( h ) ( groups 2 and 4 ) or 24 hours ( Groups 3 and 5 ) . P01375 -α ( P01375 -α ) and malondialdehyde ( MDA ) levels in the serum and retinochoroidal tissue and nuclear factor kappa-B ( NFκB ) levels in retinochoroidal tissue were determined . Histopathological examination was performed and retinochoroidal changes were scored . RESULTS : DB04933 treatment resulted in lower levels of P01375 -α , MDA , and NFκB after 12 h which became significant after 24 h . Serum P01375 -α and retinochoroidal tissue NFκB levels were similar to control animals at the 24th h of the study . DB04933 significantly reversed histopathological damage after 24 h . CONCLUSIONS : DB04933 treatment resulted in less inflammatory damage in terms of serum and retinochoroidal tissue parameters . 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 . Metaplastic regulation of the median raphe nucleus via serotonin P08908 receptor on hippocampal synaptic plasticity is associated with gender-specific emotional expression in rats . Gender differences in psychiatric disorders are considered to be associated with the serotonergic ( 5-HTergic ) system ; however the underlying mechanisms have not been clearly elucidated . In this study , possible involvement of the median raphe nucleus ( MRN ) -hippocampus 5-HTergic system in gender-specific emotional regulation was investigated , focusing on synaptic plasticity in rats . A behavioral study using a contextual fear conditioning ( Q15814 ) paradigm showed that the females exhibited low anxiety-like behavior . Extracellular 5-HT levels in the hippocampus were increased by Q15814 only in the males . Long-term potentiation ( LTP ) in the hippocampal P00915 field was suppressed after Q15814 in the males , which was mimicked by the synaptic response to MRN electrical stimulation . In the MRN , 5-HT immunoreactive cells significantly increased in the females compared with those in the males . Pretreatment with the P08908 receptor agonists tandospirone ( 10 mg/kg , i.p. ) and 8-OH DPAT ( 3 mg/kg , i.p. ) significantly suppressed LTP induction in the males . Synaptic responses to Q15814 and P08908 receptor interventions were not observed in the females . These results suggest that the metaplastic 5-HTergic mechanism via P08908 receptors in the MRN-hippocampus pathway is a key component for gender-specific emotional regulation and may be a cause of psychiatric disorders associated with vulnerability or resistance to emotional stress . [ Clinical pharmacokinetic of maraviroc ] . DB04835 ( MVC , UK-427,857 ) is the first member of a new class , the P51681 antagonists . By an original mechanism of action , maraviroc binds to the P51681 receptor in order to prevent HIV from binding and entering human cells . DB04835 ( Celsentri ) is an orally administered drug available as 150 and 300 mg film-coated tablets . The current approved daily dosage of maraviroc is 300 mg bid in combination with other antiretroviral medications . DB04835 plasma exposure is not dose proportional . After a rapid ( but moderate ) intestinal absorption , several inactive oxidized metabolites are produced via cytochrome P450 3A4 pathway . According to this liver metabolism , dosage adjustments are required when maraviroc is administered in combination with cytochrome P450 inhibitors or inducers . The potential for drug-drug interactions and the well-defined relationship between plasma concentrations and virological response suggest the usefulness of Therapeutic Drug Monitoring of maraviroc in HIV-infected patients . 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 . 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 . 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 . 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 . Antagonistic action of novel 1alpha,25-dihydroxyvitamin D3-26 , 23-lactone analogs on differentiation of human leukemia cells ( HL-60 ) induced by 1alpha,25-dihydroxyvitamin D3 . We examined the effects of two novel 1alpha,25-dihydroxyvitamin D3-26,23-lactone ( 1alpha,25-lactone ) analogues on human promyelocytic leukemia cell ( HL-60 ) differentiation using the evaluation system of the vitamin D nuclear receptor ( P11473 ) /vitamin D-responsive element ( DRE ) -mediated genomic action stimulated by 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) and its analogues . We found that the 1alpha,25-lactone analogues ( 23S ) -25-dehydro-1alpha-hydroxyvitamin-D3-26,23-lactone ( TEI-9647 ) , and ( 23R ) -25-dehydro-1alpha-hydroxyvitamin-D3-26,23-lactone ( TEI-9648 ) bound much more strongly to the P11473 than the natural ( 23S , 25R ) - DB00136 -26,23-lactone , but did not induce cell differentiation even at high concentrations ( 10(-6) M ) . Intriguingly , the differentiation of HL-60 cells induced by DB00136 was inhibited by either TEI-9647 or TEI-9648 but not by the natural lactone . In contrast , retinoic acid or 12-O-tetradecanoylphorbol-13-acetate-induced HL-60 cell differentiation was not blocked by TEI-9647 or TEI-9648 . In separate studies , TEI-9647 ( 10(-7) M ) was found to be an effective antagonist of both DB00136 ( 10(-8) M ) mediated induction of P38936 ( P38936 , CIP1 ) in HL-60 cells and activation of the luciferase reporter assay in COS-7 cells transfected with cDNA containing the DRE of the rat DB00146 -24-hydroxylase gene and cDNA of the human P11473 . Collectively the results strongly suggest that our novel 1alpha,25-lactone analogues , TEI-9647 and TEI-9648 , are specific antagonists of 1alpha , 25(OH)2D3 action , specifically P11473 /DRE-mediated genomic action . As such , they represent the first examples of antagonists , which act on the nuclear P11473 . 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 . 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 . Xaliproden ( SR57746A ) induces P08908 receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT-1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . 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 .

MH_train_1504

MH_train_1504

MH_train_1504

interacts_with DB04905?

multiple_choice

P05305 promotes survival and chemoresistance in chronic lymphocytic leukemia B cells through P25101 receptor . The endothelin axis , comprising endothelins ( ET-1 , P20800 and P14138 ) and their receptors ( ET(A)R and ETBR ) , has emerged as relevant player in tumor growth and metastasis . Here , we investigated the involvement of ET-1/ET(A)R axis in chronic lymphocytic leukemia ( CLL ) . CLL cells expressed higher levels of ET-1 and P25101 receptor as compared to normal B cells . ET-1 peptide stimulated phosphoinositide-3-kinase and mitogen-activated protein kinase signaling pathways , improved survival and promoted proliferation of leukemic cells throughout ET(A)R triggering . Moreover , the blockade of ET(A)R by the selective antagonist BQ-123 inhibited the survival advantage acquired by CLL cells in contact with endothelial layers . We also found that blocking ET(A)R via BQ-123 interferes with P29323 phosphorylation and CLL pro-survival effect mediated by B-cell receptor ( P11274 ) activation . The pro-apoptotic effect of phosphoinositide-3-kinase δ inhibitor idelalisib and mitogen-activated protein kinase inhibitor PD98059 was decreased by the addition of ET-1 peptide . Then , ET-1 also reduced the cytotoxic effect of fludarabine on CLL cells cultured alone or co-cultured on endothelial layers . ET(A)R blockade by BQ-123 inhibited the ET-1-mediated protection against drug-induced apoptosis . Lastly , higher plasma levels of big ET-1 were detected in patients ( n = 151 ) with unfavourable prognostic factors and shorter time to first treatment . In conclusion , our data describe for the first time a role of ET-1/ET(A)R signaling in CLL pathobiology . ET-1 mediates survival , drug-resistance , and growth signals in CLL cells that can be blocked by ET(A)R inhibition . P06401 modulator DB05366 induces extracellular matrix metalloproteinase inducer in cultured human uterine leiomyoma cells . Effects of progesterone receptor modulator DB05366 on the expression of the extracellular matrix ( Q13201 ) components were examined in cultured human uterine leiomyoma and myometrial cells . Q13201 metalloproteinase inducer ( P35613 ) , matrix metalloproteinases ( MMPs ) , tissue inhibitors of MMP ( TIMPs ) and collagen levels were assessed by Western blot analysis , MMP activity assay and real-time RT-PCR . RNA interference ( RNAi ) of P35613 was performed using small interfering mRNA . In cultured leiomyoma cells , DB05366 treatment at concentrations greater than or equal to 10(-8) M significantly increased P35613 , P03956 and P22894 protein contents and P03956 , P08253 , P08254 and P14780 mRNA levels , and activity of P03956 , P08253 , P08254 and P14780 in the medium . P01033 and P16035 were significantly decreased at mRNA and protein levels by DB05366 treatment at concentrations > or =10(-7) M in these cells . DB05366 treatment decreased types I and III collagen protein contents . However , DB05366 treatment did not affect the Q13201 component expression in cultured myometrial cells . RNAi of P35613 abrogated DB05366 -mediated both induction of MMPs and reduction of TIMPs and collagens in cultured leiomyoma cells . These results suggest that DB05366 modulates the expression of P35613 , MMPs , TIMPs and collagens in cultured leiomyoma cells without comparable effects on myometrial cells . The antiproliferative effect of lidocaine on human tongue cancer cells with inhibition of the activity of epidermal growth factor receptor . Local anesthetics suppress proliferation in several cancer cells . The mechanism of the suppression , however , is unknown . Our previous study shows that lidocaine , at the level of tissue concentration under topical or local administration , has a direct inhibitory effect on the activity of epidermal growth factor receptor ( P00533 ) , which is a potential target for antiproliferation in cancer cells . Therefore , we hypothesized that lidocaine would suppress the proliferation of cancer cells through the inhibition of P00533 activity . We investigated the effects of lidocaine ( 40-4000 microM ) on proliferation of a human tongue cancer cell line , CAL27 , which has a high level of P00533 expression , and also examined the effect of lidocaine on epidermal growth factor ( P01133 ) -stimulated autophosphorylation of P00533 in CAL27 cells . A clinical concentration of lidocaine ( 400 microM ) suppressed both serum-induced and P01133 -induced proliferation of CAL27 cells and inhibited P01133 -stimulated tyrosine kinase activity of P00533 without cytotoxicity . A larger concentration of lidocaine ( 4000 microM ) showed cytotoxicity with an antiproliferative effect . We suggest that the inhibition of P01133 -stimulated P00533 activity is one of the mechanisms of the antiproliferative effect of lidocaine on CAL27 cells . DB00281 administered topically within the oral cavity for cancer pain relief may suppress the proliferation of human tongue cancer cells . Hepatobiliary transport of bile acids and organic anions . Recent studies have elucidated the mechanism and regulation of hepatic transport of bile acids and organic anions . Bile acids are taken up into hepatocytes by basolateral transporters , Na+-dependently by Na+/taurocholate cotransporting polypeptide ( Q14973 ) and Na+-independently by organic anion transporting polypeptide ( P46721 ) families . Organic anions are taken up into hepatocytes by P46721 families . These compounds are then transported in hepatocytes bound to cytosolic binders , and subjected to transport by DB00171 binding cassette ( DB01048 ) transporters at the canalicular membrane . Amidated bile acids are excreted into bile by bile salt export pump ( O95342 ) , and organic anions and bile acid sulfates and glucuronides are excreted by multidrug resistance protein 2 ( Q92887 ) . Hepatic transporters are downregulated under cholestasis in rats and humans , except for O15438 , a basolateral ABC transporter , which is upregulated and may have a role in removing bile acids and organic anions from hepatocytes to the blood under cholestatic conditions . Nuclear receptors , which bind bile acids , have been shown to regulate the expression of hepatic transporters . Farnesoid X receptor ( Q96RI1 ) , which downregulates P22680 , the rate-limiting enzyme of bile acid biosynthesis , upregulates O95342 and downregulates Q14973 . Q92887 is upregulated by both Q96RI1 and pregnane X receptor ( O75469 ) , which upregulates CYP3A . Gene expression analysis of 1,25(OH)2D3-dependent differentiation of HL-60 cells : a cDNA array study . The alterations in gene expression associated with 1,25(OH)2D3-induced differentiation of HL-60 cells were studied in order to identify potential targets for further investigation of the genetic basis of acute myeloid leukaemia . Atlas human haematology filters , including 406 genes ( Clontech ) , were used to study gene expression in response to 1,25(OH)2D3 ( concentration , 5 x 10-8 mol/l ) for 24 and 72 h . Compared with untreated cells , expression differences were found in 43 genes . Downregulated genes at both time-points were : P01589 , CMYC , P06748 , P35659 , P51825 , Q01543 , htlf , P41218 , P11274 , Q13422 , P17213 and Q12968 . Upregulated genes at both time-points were P01584 , P08571 and Q07820 . P08174 , P19256 , P14316 , P16220 , P18848 , P63000 , TIAR , P42331 , P48634 , Q06187 , RCK , EV12B and P10153 were downregulated at 24 h , while P17947 , P46734 , P62324 and P10145 were upregulated . At 72 h the upregulated genes were IL1RA , P31785 , P61073 , P22362 , P10147 , P13236 , P13501 , O00626 , P07355 , Q01151 and Q03405 . cDNA array results were confirmed on randomly selected genes using quantitative real-time polymerase chain reaction for three upregulated ( P61073 , P01584 and P08571 ) and three downregulated ( P35659 , P51825 and Q01543 ) genes . Gene expression analysis after differentiation induction may provide a tool to study the roles of P35659 , P51825 and Q01543 in cell proliferation and differentiation . To demonstrate the genes that initiate differentiation , sequential gene expression analysis has to be performed during the first 24 h of the differentiation process . Ligation of surface Ig by gut-derived antigen positively selects chicken bursal and peripheral B cells . In many mammals and birds , B cell lymphopoiesis takes place in P07902 , such as the avian bursa of Fabricius . Although P11274 expression is sufficient for bursal colonization , the role of P11274 ligation in the later stages of bursal B cell lymphopoiesis remains elusive . To address this directly , we introduced a surface Ig-related construct with defined Ag specificity containing the Ag-binding portion of a lamprey variable lymphocyte receptor specific for PE fused to a truncated chicken μ-chain ( VLR(PE)Tμ ) into developing chick embryos . VLR(PE)Tμ expression supports bursal follicle colonization , clonal expansion , and Ig V gene diversification . VLR(PE)Tμ-expressing B cells migrate to the periphery in the absence of the Ag starting from day 18 of embryogenesis . VLR(PE)Tμ-expressing B cells declined rapidly in the bursa and periphery in the absence of Ag after hatch ; however , intrabursal injection of PE prolonged survival of VLR(PE)Tμ(+) bursal and peripheral B cells . Intrabursal introduction of Ag increased emigration of short-lived LT2(+) B cells . Peripheral VLR(PE)Tμ(+) B cells were maintained following intrabursal PE application and contained both short-lived LT2(+) and long-lived LT2(-) B cells . In the chicken bursa , the later stages of B cell development occur in the presence of gut-derived Ag ; therefore , we conclude that Ag-mediated ligation of P11274 in bursal B cells acts to positively select bursal B cells into both short-lived and long-lived peripheral B cell populations . DB08932 ( Opsumit ) for the treatment of pulmonary arterial hypertension . The endothelin pathway is a key pathway for the pathogenesis of pulmonary arterial hypertension ( PAH ) . Antagonism of this pathway is recommended as initial therapy in low-risk patient with PAH to inhibit fibrosis , cell proliferation , and inflammation caused by endothelin . Prior to October 2013 , ambrisentan , a selective P25101 receptor antagonist and DB00559 , a dual P25101 /ETB antagonist , were the only currently available agents for PAH targeting the endothelin pathway . Based on the results of the SERAPHIN trial , macitentan ( brand name Opsumit® ) , a new P25101 /ETB antagonist , has been US FDA approved to delay disease progression and reduce hospitalizations for PAH . SERAPHIN is the first ERA trial to use an event-driven strategy with a composite primary end point of morbidity or mortality . Previous trials have focused on short-term outcomes , such as improved 6-min walk distance and WHO functional class . 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 . 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 . Increased serum levels of neutrophil gelatinase-associated lipocalin in patients with essential thrombocythemia and polycythemia vera . Neutrophil gelatinaase-associated lipocalin ( P80188 ) is a glycoprotein bound with matrix metalloproteinase-9 ( P14780 ) in human neutrophils , and elevated tissue P80188 expression has been documented in different infectious and inflammatory conditions . Recent evidence suggests that P80188 expression is induced in many types of human cancer . Moreover , P80188 is required for P11274 - P00519 -induced tumorigenesis . The aim of the present study was to measure serum levels of P80188 in patients with essential thrombocythemia ( ET ) and polycythemia vera ( PV ) . We also evaluated P80188 levels in patients with ET and PV with and without thrombotic events , to explore a possible correlation of P80188 with platelet and leukocyte activation , and in patients with sepsis . Serum P80188 levels in the study population were significantly higher than in healthy adults and in subjects with sepsis . A correlation between P80188 and the number of white cells and neutrophils was found in patients with PV and ET . P80188 serum levels were not different depending on the presence or not of the O60674 mutation , and a mutant allele dosage effect was not observed for P80188 levels . Patients with PV and ET with thrombosis did not have significantly higher levels of P80188 . We were unable to demonstrate a significant association between serum P80188 levels and CD11b or CD62 expression . In conclusion , our study reports evidence demonstrating that increased levels of P80188 appear to be a characteristic of patients with PV and ET . Development of imatinib and dasatinib resistance : dynamics of expression of drug transporters P08183 , P33527 , Q9UNQ0 , Q14764 , and O15245 . About 20 % of patients with chronic myeloid leukemia ( CML ) do not respond to treatment with imatinib either initially or because of acquired resistance . To study the development of CML drug resistance , an in vitro experimental system comprising cell lines with different resistance levels was established by exposing K562 cells to increasing concentrations of imatinib and dasatinib anticancer agents . The mRNA levels of P11274 - P00519 and of genes involved in drug transport or redistribution ( P08183 , P33527 , O15438 , Q9UNQ0 , Q14764 , and O15245 ) were measured and the P00519 kinase domain sequenced . Results excluded P11274 - P00519 overexpression and mutations as relevant resistance mechanisms . Most studied transporters were overexpressed in the majority of resistant cell lines . Their expression pattern was dynamic : varying with resistance level and chronic drug exposure . Studied efflux transporters may have an important role at the initial stages of resistance , but after prolonged exposure and for higher doses of drugs other mechanisms might take place . 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 . 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 . Differential actions of vasopeptidase inhibition versus angiotensin-converting enzyme inhibition on diuretic therapy in experimental congestive heart failure . BACKGROUND : DB00886 ( OMA ) , a vasopeptidase inhibitor , simultaneously inhibits angiotensin-converting enzyme ( P12821 ) and neutral endopeptidase , which degrades vasodilatory factors ( eg , P35318 ) and natriuretic peptides . Based on the beneficial cardiorenal and humoral properties of the natriuretic peptides , we hypothesized that an acute vasopeptidase inhibitor with or without diuretic would result in more favorable cardiorenal and hormonal actions than P12821 inhibition plus diuretic ( ACEI+D ) in congestive heart failure . METHODS AND RESULTS : We compared the actions of OMA alone and with diuretic ( OMA+D ) to ACEI+D in a model of pacing-induced congestive heart failure . OMA+D decreased pulmonary arterial and pulmonary capillary wedge pressures to a greater level than OMA alone or ACEI+D . Glomerular filtration rate was lower with ACEI+D than with either OMA group . Plasma renin activity and aldosterone immediately increased with ACEI+D , whereas OMA+D resulted in higher plasma renin activity and a delayed increase in aldosterone . OMA alone did not increase plasma renin activity and aldosterone , but resulted in a sustained increase in plasma adrenomedullin , with higher urinary atrial natriuretic peptide , adrenomedullin , and cGMP excretions than with ACEI+D . CONCLUSIONS : Acute administration of OMA with or without diuretic results in more favorable cardiorenal and humoral responses in experimental congestive heart failure than does ACEI+D . There is no acute activation of renin and aldosterone with OMA alone such as occurs with ACEI+D and OMA+D . Thus , OMA with or without a diuretic possesses beneficial cardiorenal and humoral actions comparable to those observed with ACEI+D that can be explained by potentiation of natriuretic peptides . A Case Study of Monozygotic Twins Apparently Homozygous for a Novel Variant of UDP-Galactose 4'-epimerase ( Q14376 ) : A Complex Case of Variant Q14376 . Epimerase deficiency galactosemia is an autosomal recessive disorder that results from partial impairment of DB03501 4'-epimerase ( Q14376 ) , the third enzyme in the Leloir pathway of galactose metabolism . Clinical severity of epimerase deficiency ranges from potentially lethal to apparently benign , likely reflecting the extent of Q14376 enzyme impairment , among other factors . We report here a case study of monozygotic twins identified by newborn screening with elevated total galactose and normal galactose-1P uridylyltransferase ( P07902 ) . Follow-up testing revealed partial impairment of Q14376 in hemolysates but near-normal activity in lymphoblasts ; molecular testing identified a missense substitution , R220W , apparently in the homozygous state . The twins were treated with dietary galactose restriction for the first 18 months of life . During this time , independent testing revealed concurrent diagnoses of Williams Syndrome in both twins , and cytomegalovirus ( CMV ) infection in one . Expression studies of R220W-hGALE in a null-background strain of Saccharomyces cerevisiae demonstrated a very limited impairment of V ( max ) for DB03501 ( UDP-Gal ) and K ( m ) for UDP-N-acetylgalactosamine ( UDP-GalNAc ) , but a galactose challenge in vivo failed to uncover any evidence of impaired Leloir function . Similarly , both twins demonstrated normal hemolysate galactose-1-phosphate ( Gal-1P ) levels following normalization of their diets at 18 months of age . While these studies can not rule out a negative consequence from some cryptic Q14376 impairment in a specific tissue or developmental stage , they suggest that the substitution , R220W , is mild to neutral , so that any Q14376 impairment in these twins is likely to be peripheral and therefore unlikely to be the cause of the negative outcomes observed . 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 . 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 . Inhibition of the invasion capacity of carcinoma cells by DB05476 , a novel synthetic inhibitor of the urokinase-type plasminogen activator system . The overall survival rate of patients suffering from carcinomas has remained poor and nearly unchanged over the last decades . This is mainly due to the so-called minimal residual disease , i.e. , remaining tumor cells that overcome surgery and/or radiotherapy and are the cause of locoregional and distant metastases . To metastasize , tumor cells take advantage of proteases to invade and remodel surrounding tissues . Here , we analyzed the efficiency of DB05476 , a novel 3-amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 expression was characterized in different carcinoma cell lines , including SCCHN , breast and cervical carcinoma . Thereafter , the invasive potential of these cell lines was determined using Matrigel invasion chambers and a spheroid cocultivation model with human fibroblasts . Q03405 expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 . A decrease of tumor cell invasion by up to 50 % was achieved in both models with the SCCHN line FaDu and the cervical carcinoma line HeLa after treatment with DB05476 . Thus , our results demonstrate the potential of DB05476 in vitro as a promising adjuvant antimetastatic therapy of carcinomas . Personalized medicine and pharmacogenetic biomarkers : progress in molecular oncology testing . In the field of oncology , clinical molecular diagnostics and biomarker discoveries are constantly advancing as the intricate molecular mechanisms that transform a normal cell into an aberrant state in concert with the dysregulation of alternative complementary pathways are increasingly understood . Progress in biomarker technology , coupled with the companion clinical diagnostic laboratory tests , continue to advance this field , where individualized and customized treatment appropriate for each individual patient define the standard of care . Here , we discuss the current commonly used predictive pharmacogenetic biomarkers in clinical oncology molecular testing : P15056 V600E for vemurafenib in melanoma ; Q9HC35 - Q9UM73 for crizotinib and P00533 for erlotinib and gefitinib in non-small-cell lung cancer ; P01116 against the use of cetuximab and panitumumab in colorectal cancer ; P04626 ( P04626 /neu ) for trastuzumab in breast cancer ; P11274 - P00519 for tyrosine kinase inhibitors in chronic myeloid leukemia ; and P29590 /RARα for all-trans-retinoic acid and arsenic trioxide treatment for acute promyelocytic leukemia . Activation of P42229 confers imatinib resistance on leukemic cells through the transcription of O14746 and P08183 . We used two imatinib resistant cell lines , K562- P35318 cells , which over-express P-glycoprotein ( a product of the P08183 gene , more commonly known as P08183 ) , and K562-hTERT cells , which over-express the telomerase reverse transcriptase ( O14746 ) , as models to show that the acquisition of multidrug resistance in CML is associated with the enhanced phosphorylation of signal transducer and activator of transcription 5 ( P42229 ) . The induction of P-glycoprotein expression that occurred in response to adriamycin treatment was accompanied by increased phosphorylation of P11274 - P00519 and P42229 , as well as increased telomerase protein expression . Intriguingly , a ChIP assay using an anti- P42229 antibody revealed direct binding of P42229 to the promoter regions of both the human O14746 gene and the P08183 gene in K562- P35318 cells . Conversely , silencing of endogenous P42229 expression by siRNA significantly reduced both the expression of P-glycoprotein and telomerase activity and resulted in the recovery of the imatinib sensitivity of K562- P35318 cells . These findings indicate a critical role for P42229 in the induction of P-glycoprotein and in the modulation of telomerase activity in drug-resistant CML cells . Furthermore , primary leukemic cells obtained from patients in blast crisis showed increased levels of phospho- P42229 , P-glycoprotein and telomerase . In contrast , none of these proteins were detectable in the cells obtained from patients in the chronic phase . Together , these findings indicate a novel mechanism that contributes toward multidrug resistance involving P42229 as a sensor for cytotoxic drugs in CML patients .

MH_train_1505

MH_train_1505

MH_train_1505

interacts_with DB00758?

multiple_choice

P00797 -angiotensin-aldosterone system in brain infarction and vascular death . The renin-angiotensin-aldosterone system has functions that may contribute to brain infarction ( BI ) . In 459 matched pairs of white patients and control subjects , we measured plasma angiotensin-converting enzyme ( P12821 ) levels , seven polymorphisms ( angiotensinogen T174M and M235T , P12821 I/D and 4656 2/3CT repeat [ rpt ] , angiotensin II type 1 receptor A1166C and A153G , and aldosterone synthase P19099 ) , and evaluated 5-year poststroke mortality . Mean plasma P12821 levels ( +/-standard error ) were significantly greater in patients than control subjects ( 37.5 +/- 0.9 vs 33.9 +/- 0.9 ) , in patients with lacunar stroke , and in patients with no previous vascular ( cerebrovascular or cardiovascular ) history . The risk for BI increased with tertiles of plasma P12821 , without an interaction with hypertension . After adjustments , the association disappeared except among patients with cardioembolic BI and those without previous vascular events . Among the polymorphisms , there was a weak association of BI with angiotensin II type 1 receptor 1166C , a weak protective effect with angiotensinogen 174M , and a strong association of angiotensinogen 235T with 5-year vascular mortality . These results suggest that renin-angiotensin-aldosterone system activity and genes contribute to cerebrovascular disease and poststroke vascular death in white patients . 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 . Novel 3,4-diarylpyrazolines as potent cannabinoid P21554 receptor antagonists with lower lipophilicity . Novel 3,4-diarylpyrazolines 1 as potent P21554 receptor antagonists with lipophilicity lower than that of DB05077 are described . The key change is the replacement of the arylsulfonyl group in the original series by a dialkylaminosulfonyl moiety . The absolute configuration ( 4S ) of eutomer 24 was established by X-ray diffraction analysis and 24 showed a close molecular fit with rimonabant in a P21554 receptor-based model . Compound 17 exhibited the highest P21554 receptor affinity ( Ki = 24 nM ) in this series , as well as very potent P21554 antagonistic activity ( pA2 = 8.8 ) and a high P21554 /CB2 subtype selectivity ( approximately 147-fold ) . 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 . 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 . 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 . P19099 inhibitors in cardiovascular and renal diseases . DB04630 is involved in various cardiovascular pathologies , including hypertension , heart failure , atherosclerosis and fibrosis . P08235 ( MR ) -dependent and -independent , genomic and non-genomic processes mediate its complex effects . Spironolactone and eplerenone , both MR antagonists , are the only commercially available compounds targeting directly the actions of aldosterone . However , due to the poor selectivity ( spironolactone ) , low potency ( eplerenone ) and the fact that only MR-dependent effects of aldosterone can be inhibited , these drugs have limited clinical use . An attractive approach to abolish potentially all of aldosterone-mediated pathologies is the inhibition of aldosterone synthase . This review summarizes current knowledge on the complex effects mediated by aldosterone , potential advantages and disadvantages of aldosterone inhibition and novel directions in the development of aldosterone synthase inhibitors . 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 . Increase in intracellular Cl- concentration by DB02527 - and Ca2+-dependent stimulation of M1 collecting duct cells . In the lungs of cystic fibrosis ( CF ) patients , mutations of the cystic fibrosis transmembrane conductance regulator ( P13569 ) lead to defective Cl- secretion and hyperabsorption of electrolytes . This may be a an important cause for the defective mucociliary clearance in CF lungs . Previous studies have suggested that inhibition of ENaC during activation of P13569 or by purinergic stimulation could be related to an increase in the intracellular [Cl-]i . This was examined in the present study using cultured mouse M1 collecting duct cells transfected with the chloride-sensitive enhanced yellow fluorescent protein YFP(V163S) . Calibration experiments showed a linear decrease of YFP fluorescence intensity with increasing [Cl-]i ( 0-100 mM ) . Activation of P13569 by isobutyl-1-methylxanthine ( DB07954 , 100 microM ) and forskolin ( 2 microM ) increased [Cl-]i by 9.6+/-1.5 mM ( n=35 ) . Similarly , DB00171 ( 100 microM ) increased [Cl-]i transiently by 9.5+/-2.2 mM ( n=17 ) . The increase in [Cl-]i was reduced by the Na+/K+/2 Cl- -cortransporter-1 ( P55011 ) blocker azosemide ( 100 microM ) , the P13569 blocker DB04941 ( 50 microM ) , the blocker of Ca2+-activated Cl- channels DIDS ( 100 microM ) or the ENaC blocker amiloride ( 10 microM ) . Changes in YFP(V163S) fluorescence were not due to changes in cell volume or intracellular pH . The present data thus demonstrate an increase in [Cl-]i following stimulation with secretagogues , which could participate in the inhibition of ENaC . 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 . 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 . The Leu33Pro polymorphism in the P05106 gene does not modify P38398 /2-associated breast or ovarian cancer risks : results from a multicenter study among 15,542 P38398 and P51587 mutation carriers . Integrins containing the beta(3) subunit are key players in tumor growth and metastasis . A functional Leu33Pro polymorphism ( rs5918 ) in the beta(3) subunit of the integrin gene ( P05106 ) has previously been suggested to act as a modifier of ovarian cancer risk in Polish P38398 mutation carriers . To investigate the association further , we genotyped 9,998 P38398 and 5,544 P51587 mutation carriers from 34 studies from the Consortium of Investigators of Modifiers of P38398 /2 for the P05106 Leu33Pro polymorphism . Data were analysed within a Cox-proportional hazards framework using a retrospective likelihood approach . There was marginal evidence that the P05106 polymorphism was associated with an increased risk of ovarian cancer for P38398 mutation carriers ( per-allele Hazard Ratio ( HR ) 1.11 , 95 % CI 1.00-1.23 , p-trend 0.05 ) . However , when the original Polish study was excluded from the analysis , the polymorphism was no longer significantly associated with ovarian cancer risk ( HR 1.07 , 95 % CI 0.96-1.19 , p-trend 0.25 ) . There was no evidence of an association with ovarian cancer risk for P51587 mutation carriers ( HR 1.09 , 95 % CI 0.89-1.32 ) . The polymorphism was not associated with breast cancer risk for either P38398 or P51587 mutation carriers . The P05106 Leu33Pro polymorphism does not modify breast or ovarian cancer risk in P38398 or P51587 mutation carriers . The ' allosteric modulator ' P35240 -202676 disrupts G protein-coupled receptor function via sulphydryl-sensitive mechanisms . 1. Previous studies suggest that the thiadiazole compound P35240 -202676 ( N-(2,3-diphenyl-1,2,4-thiadiazol-5-(2H)-ylidene)methanamine ) acts as an allosteric modulator of a variety of structurally distinct G protein-coupled receptors ( GPCRs ) . It was postulated that P35240 -202676 would directly bind a structural motif in the receptor molecule common to divergent members of the GPCR family . The molecular mechanisms of such a promiscuous action , however , remain obscure . 2 . To clarify the mechanism of P35240 -202676 action , we used the functional approach of [35S]GTPgammaS autoradiography with rat brain cryostat sections together with classical membrane [35S]GTPgammaS binding assays to evaluate how the thiadiazole affects G protein activity mediated by various receptors linked to the Gi-family of G proteins . 3 . We found that in the absence of dithiotreitol ( DTT ) , P35240 -202676 ( 10(-7)-10(-5) M ) elicits nonspecific effects in the [35S]GTPgammaS-based G protein activation assays , thereby severely compromising interpretations on the compounds ability to allosterically inhibit receptor-mediated G protein activity . Such a nonspecific behaviour was fully reversed upon addition of DTT ( 1 mM ) , revealing thiol-based mechanism of action . 4 . In routine incubations containing DTT , P35240 -202676 had no effect on receptor-driven G protein activity , as assessed for adenosine A1 , alpha2-adrenergic , cannabinoid P21554 , lysophosphatidic acid Q92633 , muscarinic M2/M4 , purinergic Q9H244 or sphingosine 1-phosphate receptors , suggesting that the thiadiazole does not act as an allosteric modulator of GPCR function . 5 . 1H NMR analysis indicated that P35240 -202676 underwent structural changes after incubation with the reducing agent DTT or with brain tissue . 6 . We conclude that P35240 -202676 modulates GPCRs via thiol modification rather than via true allosteric mechanisms . 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 . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . 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 . Mammalian homologues of C. elegans P25116 are asymmetrically localized in epithelial cells and may influence their polarity . The establishment of polarity in the embryo is fundamental for the correct development of an organism [ 1 ] . The first cleavage of the Caenorhabditis elegans embryo is asymmetric with certain cytoplasmic components being distributed unequally between the daughter cells [ 2-4 ] . Using a genetic screen , Kemphues and co-workers have identified six par genes ( partition-defective ) [ 5,6 ] , which are involved in the process of asymmetric division . One of these genes encodes a highly conserved protein , P25116 , which is a serine/threonine kinase that localizes asymmetrically to the posterior part of the zygote and to those blastocysts that give rise to the germ line [ 7-9 ] . We reasoned that the mammalian homologue of P25116 ( mPAR-1 ) might be involved in the process of polarization of epithelial cells , which consist of apical and basolateral membrane domains . We found that mPAR-1 was expressed in a wide variety of epithelial tissues and cell lines and was associated with the cellular cortex . In polarized epithelial cells , mPAR-1 was asymmetrically localized to the lateral domain . A fusion protein lacking the kinase domain had the same localization as the full-length protein but its prolonged expression acted in a dominant-negative fashion : lateral adhesion of the transfected cells to neighbouring cells was diminished , resulting in the former cells being ' squeezed out ' from the monolayer . Moreover , the polarity of these cells was disturbed resulting in mislocalization of P12830 . Thus , in the C. elegans embryo and in epithelial cells , polarity appears to be governed by similar mechanisms . Anti-stress effect of astragaloside IV in immobilized mice . ETHNOPHARMACOLOGICAL RELEVANCE : Astragaloside IV , a major component extracted from the roots of Astragalus membranaceus ( AM ) , possesses anti-inflammatory , anti-oxidative , anti-fibrotic , anti-infarction and immunoregulatory effects . To clarify anti-stress effect of AM , anxiolytic and anti-inflammatory effects of 80 % ethanol extract of AM and astragaloside IV were investigated in immobilization stress model . MATERIALS AND METHODS : The mice were orally administered with AM ( 50 , 200 , and 500 mg/kg ) , astragaloside IV ( 5 , 10 , and 20 mg/kg ) and buspirone , a positive drug , 1h before immobilization treated for 2h . For anxiolytic activity assay , EPM test was performed in mice . For anti-inflammatory activity assay , serum levels of corticosterone , P05231 and P01375 -α were measured using ELISA kits . RESULTS : AM extract and astragaloside IV increased dose-dependently time spent on open arms and open arm entries in the EPM test . Anxiolytic effects of AM extract ( 500 mg/kg ) and astragaloside IV ( 20 mg/kg ) were comparable to those of buspirone ( 1 mg/kg ) . Their anxiolytic effects were blocked by WAY-100635 ( 0.5 mg/kg , i.p. ) , a P08908 receptor antagonist ( p < 0.01 ) , but not by flumazenil ( 3 mg/kg , i.p. ) and bicuculline ( 0.5 mg/kg , i.p. ) , GABAA receptor antagonists . AM extract and astragaloside IV also reduced serum levels of corticosterone , P05231 and P01375 -α dose-dependently . CONCLUSIONS : AM , particularly astragaloside IV , may ameliorate immobilized stress-induced anxiety and inflammation . Pharmacological effects of lipid-lowering drugs recapitulate with a larger amplitude the phenotypic effects of common variants within their target genes . BACKGROUND : A major expectation underlying the search for novel susceptibility genes for common diseases using genome-wide association studies ( GWAS ) is that these discoveries will lead to new drug targets . This claim has not been verified yet . Here , we tested the hypothesis that common single nucleotide polymorphisms ( SNPs ) within drug target genes are associated with the corresponding phenotypes , using a population-based GWAS dataset and lipid-lowering drugs as a test case . METHODS : We examined the association between 36 genotyped and 193 imputed SNPs within four lipid-lowering drug target genes ( P04035 , Q07869 , Q8TDS4 / Q8TDS4 and P11597 ) and four non-lipid drug target genes ( P12821 , P30556 , Q9H244 , and P51164 ) and lipid phenotypes , blood pressure , and coronary artery disease in 5635 adult participants of the Lausanne , Switzerland , CoLaus study , genotyped using the Affymetrix 500K SNP chip technology . RESULTS : The phenotypes associated with SNPs within drug target genes recapitulated to a certain extent the pharmacological effects of the drug . The amplitude of the SNP effect was about 10 times smaller than the pharmacological effect of the corresponding drug . In particular , several P11597 SNPs were associated with an elevation in HDL-cholesterol levels , yet a lower diastolic blood pressure , providing evidence that the blood pressure elevation induced by the P11597 inhibitor torcetrapib is more likely compound specific than class specific . CONCLUSION : Pharmacological modulation of lipid-lowering drug targets recapitulates , and markedly amplifies , the phenotypic effects of common SNPs within these target genes . This data provides indirect evidence that , with certain limitations , large-scale GWAS represent a new tool for the discovery and the development of innovative drugs . 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 . Protease-activated receptor regulation of Cl- secretion in Calu-3 cells requires prostaglandin release and P13569 activation . Human lung epithelial ( Calu-3 ) cells were used to investigate the effects of protease-activated receptor ( PAR ) stimulation on Cl(-) secretion . Quantitative RT-PCR ( QRT-PCR ) showed that Calu-3 cells express P25116 , -2 , and -3 receptor mRNAs , with P55085 mRNA in greatest abundance . Addition of either thrombin or the P55085 agonist peptide SLIGRL to the basolateral solution of monolayers mounted in Ussing chambers produced a rapid increase in short-circuit current ( I(sc) : thrombin , 21 +/- 2 microA ; SLIGRL , 83 +/- 22 microA ) , which returned to baseline within 5 min after stimulation . Pretreatment of monolayers with the cell-permeant Ca(2+)-chelating agent BAPTA-AM ( 50 microM ) abolished the increase in I(sc) produced by SLIGRL . When monolayers were treated with the cyclooxygenase inhibitor indomethacin ( 10 microM ) , nearly complete inhibition of both the thrombin- and SLIGRL-stimulated I(sc) was observed . In addition , basolateral treatment with the PGE(2) receptor antagonist AH-6809 ( 25 microM ) significantly inhibited the effects of SLIGRL on I(sc) . QRT-PCR revealed that Calu-3 cells express mRNAs for P13569 , the Ca(2+)-activated O15554 K(+) channel , and the P51787 K(+) channel subunit , which , in association with Q9Y6H6 , is known to be regulated by DB02527 . Stimulation with SLIGRL produced an increase in apical Cl(-) conductance that was blocked in cells expressing short hairpin RNAs designed to target P13569 . These results support the conclusion that PAR stimulation of Cl(-) secretion occurs by an indirect mechanism involving the synthesis and release of prostaglandins . In addition , PAR-stimulated Cl(-) secretion requires activation of P13569 and at least two distinct K(+) channels located in the basolateral membrane . 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 .

MH_train_1506

MH_train_1506

MH_train_1506

interacts_with DB03880?

multiple_choice

Production of prostaglandins in transgenic Arabidopsis thaliana . Plants do not naturally produce the very-long-chain polyunsaturated fatty acids that are the precursors of prostaglandins , but in previous studies Arabidopsis thaliana had been transformed sequentially with genes encoding a Δ(9)-elongase and a Δ(8)-desaturase to produce dihomo-γ-linolenic acid ( DB00154 ) and eicosatetraenoic acid ( P25101 ) , and subsequently with a gene encoding a Δ(5)-desaturase to produce arachidonic acid ( AA ) and eicosapentaenoic acid ( EPA ) . Transformation of A. thaliana with the first two genes consolidated on a single binary vector yielded transformants producing high levels of DB00154 , and these plants were further transformed with mouse prostaglandin H synthase ( PGH ) genes to produce prostaglandins . Mouse P23219 and P35354 cDNAs were amplified for expression as three isoforms : P23219 ( complete coding sequence with signal peptide ) , P23219 -Ma ( mature P23219 sequence , without signal peptide ) and P35354 ( complete coding sequence with signal peptide ) . P23219 transformants showed the highest activity , followed by P35354 transformants , whereas removal of the signal peptide resulted in almost complete loss of P23219 activity . In order to produce a physiologically active prostaglandin , the Trypanosoma brucei prostaglandin F synthase gene was combined with the mouse P23219 gene and the Mortierella alpina Δ(5)-desaturase on a binary vector . Transformation of DB00154 -producing A. thaliana with this construct yielded transformants that successfully produced prostaglandin F . 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 . Endocellular polyamine availability modulates epithelial-to-mesenchymal transition and unfolded protein response in MDCK cells . Epithelial-to-mesenchymal transition ( EMT ) is involved in embryonic development as well as in several pathological conditions . Literature indicates that polyamine availability may affect transcription of c-myc , matrix metalloproteinase (MMP)1 , P08253 , TGFbeta(1) , and collagen type I mRNA . The aim of this study was to elucidate polyamines role in EMT in vitro . Madin-Darby canine kidney ( MDCK ) cells were subjected to experimental manipulation of intracellular levels of polyamines . Acquisition of mesenchymal phenotype was evaluated by means of immunofluorescence , western blots , and zymograms . MDCK cells were then subjected to 2D gel proteomic study and incorporation of a biotinilated polyamine ( Q03001 ) . Polyamine endocellular availability modulated EMT process . Polyamine-depleted cells treated with TGFbeta(1) showed enhanced EMT with a marked decrease of P12830 expression at plasma membrane level and an increased expression of mesenchymal markers such as fibronectin and alpha-smooth muscle actin . Polyamine-depleted cells showed a twofold increased expression of the rough endoplasmic reticulum ( ER ) -stress proteins P11021 , P14625 , and HSP90 alpha/beta in 2D gels . The latter data were confirmed by western blot analysis . Administration of Q03001 showed that polyamines are covalently linked , within the cell , to ER-stress proteins . Intracellular polyamine availability affects EMT in MDCK cells possibly through the modulation of ER-stress protein homeostasis . 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 . 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 . Type 1 insulin-like growth factor regulates P50281 synthesis and tumor invasion via PI 3-kinase/Akt signaling . The membrane type 1 matrix metalloproteinase ( P50281 ) has been identified as a major activator of P08253 - a process involving the formation of a trimolecular complex with P16035 . We previously identified the P08069 as a positive regulator of P08253 synthesis . Here , we investigated the role of IGF-IR in the regulation of P50281 . Highly invasive Lewis lung carcinoma subline H-59 cells express P50281 and utilize it to activate their major extracellular matrix degrading proteinase- P08253 . These cells were transiently transfected with a plasmid vector expressing a luciferase reporter gene downstream of the mouse P50281 promoter . P05019 treatment increased luciferase activity in the transfected cells by up to 10-fold and augmented endogenous P50281 mRNA and protein synthesis by up to 2-3-fold , relative to controls . P50281 induction and invasion were blocked by the PI 3-kinase inhibitors LY294002 and wortmannin and by rapamycin , but not by the MEK inhibitor PD98059 . Overexpression of a dominant negative Akt mutant or of the tumor suppressor phosphatase and tensin homologue , P60484 , in these cells also caused a significant reduction in P50281 expression and invasion . The results demonstrate that IGF-IR controls tumor cell invasion by coordinately regulating P08253 expression and its P50281 -mediated activation and identify PI 3-kinase/Akt/ P42345 signaling as critical to this regulation . [ 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 . 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 . 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 . 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 . A new murine P01730 + T cell subset with an unrestricted cytokine profile . P01730 + T cell clones were derived from mice immunized to DB05299 to characterize the cytokine profiles of newly isolated clones . Surprisingly , several of the clones had an unrestricted profile , producing P60568 , P08700 , P05112 , P01579 , and P01375 after either Con A or Ag stimulation . The coproduction of P60568 and P05112 was confirmed at the mRNA level . Subclones were derived which contained RNA transcripts for , as well as secreted , both P60568 and P05112 thus confirming the clonality of the original T cell clones . P01730 + T cell clones that expressed an unrestricted cytokine profile upon Con A stimulation were also isolated from mice immunized to other Ag ( hen egg lysozyme , OVA , or type II collagen ) . These data indicate that P01730 + T cell clones newly isolated from immunized mice do not necessarily segregate into the Th1 and Th2 subsets . We propose this new murine P01730 + cell subset with an unrestricted pattern of cytokine production be called Th0 . 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 . Inhibition of p38alpha MAPK disrupts the pathological loop of proinflammatory factor production in the myelodysplastic syndrome bone marrow microenvironment . Myelodysplastic syndromes ( P43034 ) are common causes of ineffective hematopoiesis and cytopenias in the elderly . Various myelosuppressive and proinflammatory cytokines have been implicated in the high rates of apoptosis and hematopoietic suppression seen in P43034 . We have previously shown that p38 MAPK is overactivated in P43034 hematopoietic progenitors , which led to current clinical studies of the selective p38alpha inhibitor , DB05412 , in this disease . We now demonstrate that the myelosuppressive cytokines TNFalpha and IL-1beta are secreted by bone marrow ( BM ) cells in a p38 MAPK-dependent manner . Their secretion is stimulated by paracrine interactions between BM stromal and mononuclear cells and cytokine induction correlates with P28906 + stem cell apoptosis in an inflammation-simulated in vitro bone marrow microenvironment . Treatment with DB05412 inhibits P01375 secretion in primary P43034 bone marrow cells and protects cytogenetically normal progenitors from apoptosis ex vivo . Furthermore , p38 inhibition diminishes the expression of TNFalpha or IL-1beta-induced proinflammatory chemokines in BM stromal cells . These data indicate that p38 inhibition has anti-inflammatory effects on the bone marrow microenvironment that complements its cytoprotective effect on progenitor survival . These findings support clinical investigation of p38alpha as a potential therapeutic target in P43034 and other related diseases characterised by inflammatory bone marrow failure . P43220 agonist may activate pancreatic stellate cells to induce rat pancreatic tissue lesion . OBJECTIVE : To explore the mechanism of P43220 agonist-induced rat pancreatic tissue lesion . METHODS : Thirty SD male rats were divided into three groups , namely P43220 agonist experimental group , diabetes-model experimental group and control group . Diabetes-model rats were induced by streptozotocin and high-sugar high-fat diet . P43220 agonist group and diabetes-model group were administered with P43220 agonist in dose 5 μg/kg each time , twice a day . After 10 weeks of treatment , the amount of matrix metalloproteinase ( MMP ) -2 and P14780 , and expression of α-smooth muscle actin ( α-SMA ) and type III collagen protein in pancreatic tissue were measured . RESULTS : The amount of P08253 and P14780 in P43220 agonist group and diabetes-model group were significantly higher than the control group . Compared with the P43220 agonist group , the diabetic model group had more severe pathological changes of pancreatic tissue interstitial edema , inflammatory cell infiltration , glandular atrophy and fibrosis , and significantly increased pancreatic tissue P08253 and P14780 levels , significantly increased α-SMA and collagen III-positive cell counts , all the differences were statistically significant . α-SMA and type III collagen were expressed in all parts of the lesions of P43220 agonist group and diabetes-model group . α-SMA can only be observed in the vessel wall in control group , however , in the other two groups , α-SMA can also be observed in pancreatic acinar cell interstitia , in addition to vessel wall . CONCLUSIONS : Long-term subcutaneous P43220 agonist injection may activate pancreatic stellate cells , causing the expression of α-SMA and collagen III and the amount of P08253 and P14780 in pancreatic acinar cell interstitial significantly increasing , and thus inducing chronic inflammatory change . The clinical implications of increased cyclophilin A levels in patients with acute coronary syndromes . BACKGROUND : P62937 is a secreted molecule that has a physiological and pathological role in cardiovascular diseases . However , limited information is available on the relationship between cyclophilin A concentration and acute coronary syndromes ( ACS ) . We investigated whether cyclophilin A concentration is related to the stability of coronary atherosclerotic plaque in patients with ACS . METHODS : This study included normal controls ( n=50 ) , patients with stable angina ( SA ) ( n=60 ) and patients with ACS , including unstable angina ( UA ) ( n=60 ) and acute myocardial infarction ( AMI ) ( n=90 ) . Serum soluble cyclophilin A , matrix metalloproteinase 9 ( P14780 ) , P08254 and P02741 concentrations ( CRP ) were measured . All coronary stenosis were assessed by angiographic coronary stenosis morphology . RESULTS : Serum cyclophilin A concentration in ACS ( UA and AMI ) subjects were significantly higher than those in patients with SA and controls ( p < 0.05 ) . Serum cyclophilin A correlated positively with serum P08254 and P14780 and CRP in ACS patients ( r(1)=0.69 , r(2)=0.52 , r(3)=0.49 p < 0.0001 ) , but not in control . Furthermore , the increased cyclophilin A concentrations was associated with the number of complex coronary stenoses ( r(1)=0.63 , p < 0.0001 ) , but not smooth lesions or stenosis severity , in coronary artery disease patients . Logistic regression analysis also demonstrated that serum cyclophilin A concentration was an independent predictor factor for ACS ( OR , 2.721 , 95 % CI 1.563-4.042 , p=0.001 ) . CONCLUSION : Patients with ACS showed that increased concentrations of cyclophilin A may be a valuable marker for predicting the severity of ACS . Altered expression of inflammation-related genes in human carotid atherosclerotic plaques . OBJECTIVE : Inflammation is a pivotal process in atherosclerosis development and progression , but the underlying molecular mechanisms remain largely obscure . We have conducted an extensive expression study of atherosclerotic plaques to identify the inflammatory pathways involved in atherosclerosis . METHODS : We studied 11 human carotid plaques , their respective adjacent regions and 7 control arteries from different subjects . Expression of 92 genes was studied by TaqMan low-density array human inflammation panel . Human aortic endothelial and smooth muscle cells were used for in vitro experiments . RESULTS : The mRNA levels of 44/92 genes ( 48 % ) differed significantly between the tissues examined ( 13 up-regulated and 31 down-regulated ) . Dysregulated genes encode molecules belonging to different functional classes although most of them encode enzymes involved in the eicosanoid synthesis pathway . The expression of Q16647 and P43119 genes was decreased in human aortic endothelial and smooth muscle cells stimulated with oxLDL and P01375 -α . CONCLUSIONS : This study not only reveals several dysregulated genes in human lesions but also focuses the role played by the genes involved in the eicosanoid synthesis pathway during atherosclerotic development . The decrease of Q16647 and P43119 expression after oxLDL treatment mirrors the decreased mRNA levels in atherosclerotic lesions versus control arteries , which suggests that oxidation is important for Q16647 and P43119 regulation in human vessel cells during atherosclerosis development . DB03758 -sensitive peptide binding to the N-terminal portion of P14625 . P14625 is a molecular chaperone that carries immunologically relevant peptides from cell to cell , transferring them to major histocompatibility proteins for presentation to T cells . Here we examine the binding of several peptides to recombinant P14625 and study the regulation and site of peptide binding . We show that P14625 contains a peptide-binding site in its N-terminal 355 amino acids . A number of peptides bind to this site with low on- and off-rates and with specificity that is distinct from that of another endoplasmic reticulum chaperone , P11021 / P11021 . Binding to the N-terminal fragment is sufficient to account for the peptide binding activity of the entire molecule . Peptide binding is inhibited by radicicol , a known inhibitor of the chaperone activities of HSP90-family proteins . However , the peptide-binding site is distinct from the radicicol-binding pocket , because both can bind to the N-terminal fragment simultaneously . Furthermore , peptide binding does not cause the same conformational change as does binding of radicicol . When the latter binds to the N-terminal domain , it induces a conformational change in the downstream , acidic domain of P14625 , as measured by altered gel mobility and loss of an antibody epitope . These results relate the peptide-binding activity of P14625 to its other function as a chaperone .

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interacts_with DB00623?

multiple_choice

DB05258 receptor 2 expression by peripheral blood monocytes in patients with a high viral load of hepatitis C virus genotype 1 showing substitution of amino Acid 70 in the core region . BACKGROUND/AIM : When patients with chronic hepatitis C ( CHC ) are treated with interferon ( IFN ) -based therapy , achieving serum HCV-RNA negativity by week 12 ( early viral response , EVR ) is an important predictor of a sustained virologic response . The aim of this study was to clarify whether changes in IFN-alpha receptor 2 ( P17181 -2 ) expression by peripheral blood monocytes ( Mo ) and the EVR rate differed between patients with genotype 1b and a high viral load showing substitution of amino acid 70 in the core region of HCV ( mutant , n = 20 ) and patients without this substitution ( wild , n = 23 ) . PATIENTS AND METHODS : Forty-three CHC patients were studied , and received pegylated IFN plus ribavirin . P17181 -2 expression by Mo was determined using flow cytometry to measure the mean fluorescence intensity ( MFI ) before and up to 28 days after starting therapy . RESULTS : The EVR rate of the mutant group was significantly lower than that of the wild group ( 35 vs.70 % ) . The MFI of Mo was significantly higher in the wild group than in the mutant group before and also 3 , 7 , and 28 days after starting therapy . CONCLUSIONS : Mutation of HCV was related to lower P17181 -2 expression by Mo before and after starting therapy . Niflumic acid and MSI-2216 reduce P01375 -induced mucin expression in human airway mucosa . BACKGROUND : Human chloride channel , calcium-activated 1 ( A8K7I4 ) has been shown to induce mucin ( MUC ) gene expression and mucus production in bronchial epithelial cells . Objective To investigate whether blocking A8K7I4 decreases mucus production . METHODS : Expression of A8K7I4 and mucus was stimulated with P01375 in human upper airway mucosal explant tissue . P98088 mRNA and mucus protein expression was blocked by inhibiting A8K7I4 by using channel blockers ( niflumic acid [ DB04552 ] and MSI-2216 ) without and with P01375 stimulation . Expression of P98088 , A8K7I4 , and P35354 mRNA was quantified by using real-time PCR . Mucus protein was assessed by periodic acid Schiff staining . Laser capture microdissection was performed to quantify A8K7I4 and P98088 mRNA expression in epithelial cells derived from mucosal explant tissue . RESULTS : P01375 significantly increased P98088 and A8K7I4 mRNA as well as mucus and A8K7I4 protein expression in the mucosal explant tissue ( P < .05 ) . Inhibition of A8K7I4 with DB04552 or MSI-2216 showed a significant dose-dependent reduction of mucus production for both blockers in the mucosal explant tissue ( P < .05 ) . P98088 mRNA was also decreased by both blockers in the whole mucosal tissue and in laser-captured mucosa epithelial cells . CONCLUSIONS : Unstimulated and P01375 -induced mucin expression could be decreased by DB04552 and MSI-2216 . Inhibiting A8K7I4 may be a potential new approach to reduce mucus overproduction . Neuropeptide Y induces ischemic angiogenesis and restores function of ischemic skeletal muscles . Previously we showed that neuropeptide Y ( P01303 ) , a sympathetic vasoconstrictor neurotransmitter , stimulates endothelial cell migration , proliferation , and differentiation in vitro . Here , we report on P01303 's actions , receptors , and mediators in ischemic angiogenesis . In rats , hindlimb ischemia stimulates sympathetic P01303 release ( attenuated by lumbar sympathectomy ) and upregulates P01303 - P28062 ( P28062 ) receptor and a peptidase forming P28062 /Y5-selective agonist . Exogenous P01303 at physiological concentrations also induces Q15761 , stimulates neovascularization , and restores ischemic muscle blood flow and performance . P01303 -mediated ischemic angiogenesis is not prevented by a selective Q03519 receptor antagonist but is reduced in P28062 (-/-) mice . Nonischemic muscle vascularity is also lower in P28062 (-/-) mice , whereas it is increased in P01303 -overexpressing rats compared with their WT controls . Ex vivo , P01303 -induced aortic sprouting is markedly reduced in P28062 (-/-) aortas and spontaneous sprouting is severely impaired in P01303 (-/-) mice . P01303 -mediated aortic sprouting , but not cell migration/proliferation , is blocked by an antifetal liver kinase 1 antibody and abolished in mice null for P29474 . Thus , P01303 mediates neurogenic ischemic angiogenesis at physiological concentrations by activating P28062 /Y5 receptors and P29474 , in part due to release of P15692 . P01303 's effectiveness in revascularization and restoring function of ischemic tissue suggests its therapeutic potential in ischemic conditions . 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 . 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 . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . Effects of gonadoliberin analogue triptorelin on the pituitary-testicular complex in neonatal rats . DB06825 , a synthetic analogue of neurohormone gonadoliberin ( gonadotropin-releasing hormone , DB00644 ) administered daily to rats on postnatal days 5-7 suppressed the expression of P30968 in the pituitary gland , but did not change functioning of the pituitary-testicular complex . Administration of triptorelin on postnatal days 12-14 ( i.e. during the formation of pulsatile pattern of DB00644 secretion and increasing levels of its mRNA receptor in the pituitary gland ) had no effect on receptor expression , but increased the levels of luteinizing hormone mRNA in the pituitary gland and the weight of testes . At that time , blood levels of testosterone were lowered , which indicated disturbed pulsatile pattern of DB00644 secretion . Involvement of cyclin-dependent kinase activities in CD437-induced apoptosis . A novel synthetic retinoid , 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid ( CD437 ) , is a selective ligand of the RARgamma nuclear receptor . We examined the in vitro effects of CD437 and found that CD437 induces S phase arrest within 24 to 48 h , followed by cell death , in the p53-negative Hep3B and the p53-positive HepG2 human hepatoma cell lines . Based on observations of cellular and nuclear fragmentation , chromatin condensation , and DNA fragmentation , the CD437-mediated cell-killing effect appears to be due to apoptosis . On morphological examination , a number of CD437-treated cells were found to have increased 5- to 10-fold in size and persisted as single giant cells without cell division , while the remainder underwent nuclear division ( multiple nuclei ) but were unable to complete cytokinesis , and finally all died by apoptosis . In HepG2 cells that possessed wild-type p53 , CD437-induced S phase arrest and apoptosis were accompanied by the up-regulation of cyclin A , cyclin B , p53 , P38936 (CIP1/Waf1) , Bad , and Bcl-Xs proteins and by a decrease in Bcl-2 protein levels . In Hep3B cells , CD437-mediated S phase arrest and apoptosis were also associated with a concomitant up-regulation of cyclin A , cyclin B , Bad , and Bcl-Xs . However , Hep3B cells did not express p53 or Bcl-2 messages . DB02116 and roscovitine , the potent p34(cdc2) and P24941 inhibitors , effectively blocked CD437-mediated cyclin A- and B-dependent kinase activation and prevented CD437-induced cell death . Furthermore , antisense oligonucleotide complementary to cyclin A and B mRNA significantly rescued CD437-induced apoptosis . These findings suggest that activation of cyclin A- and B-dependent kinases is a critical determinant of apoptotic death mediated by CD437 . Glucocorticoids regulate calcineurin-dependent trans-activating pathways for interleukin-2 gene transcription in human T lymphocytes . Glucocorticoids ( GC ) inhibit P60568 gene transcription by interfering with the binding of the nuclear factor activator protein-1 on the P60568 promoter . Calcineurin , a Ca2+/calmodulin-dependent protein phosphatase , is an essential component of the T cell antigen receptor signal transduction pathway leading to P60568 gene transcription . Therefore , we have asked whether this phosphatase may also be regulated by GC . Jurkat T cells were cotransfected with plasmids containing the intact P60568 promoter or its NF-AT and P14859 motifs , and a deletion mutant ( delta P62158 -AI ) of calcineurin known to have Ca(2+)-independent constitutive phosphatase activity . Cotransfection of P60568 promoter with delta P62158 -AI allowed the activation of P60568 promoter in the presence of phorbol ester alone . Under these conditions dexamethasone ( DB00514 ; 10(-6) M ) inhibited P60568 promoter activation by 50-60 % . The inhibitory effect of DB00514 was specific , as demonstrated by experiments using an unrelated promoter ( simian virus 40 ) and estradiol . Furthermore , it was completely reversed in the presence of excess amounts of the glucocorticoid antagonist RU 486 , which suggests that it is mediated through the glucocorticoid receptor . Overexpression of calcineurin via delta P62158 -AI in Jurkat cells decreased their apparent sensitivity to DB00514 ( approximately 5-fold increase in IC50 ) . Similar results were obtained with the NF-AT and P14859 constructs , which are also known to be activated by calcineurin . Thus , in addition to their known inhibitory effects on activator protein-1 , GC also inhibit calcineurin-dependent pathways for T cell 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 . Hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in the rat . Growth hormone-releasing peptides ( Q92847 ) and ghrelin are synthetic and natural ligands of growth hormone secretagogue receptor ( Q92847 ) respectively and are shown to exert protective actions on cardiac dysfunction . Because ghrelin has been reported to inhibit proinflammatory responses in human endothelium and Q92847 has been identified in blood vessels , we hypothesized that Q92847 could alleviate the development of atherosclerosis ( As ) . Atherosclearosis was induced by a short period ( 4 days ) of vitamin D(3) and chronic ( three months ) intragastric feeding of high fat emulsion ( containing 0.5 % propylthiouracil ) in adult SD rats . Some As rats received chronic hexarelin ( a variant of Q92847 ) injection ( SC P55957 , 30 days ) and normal rats received placebo as control . Significant atherosclerosis developed in animals fed with the emulsion . Serum total cholesterol and LDL-c increased , and HDL-c and aortic nitric oxide ( NO ) decreased significantly in As group . Hexarelin suppressed the formation of atherosclerotic plaques and neointima , partially reversed serum HDL-c/LDL-c ratio and increased the levels of serum NO and aortic mRNAs of P29474 , Q92847 and P16671 in As rats . Hexarelin also decreased [ (3)H ] -TdR incorporation in cultured vascular smooth muscle cell ( VSMC ) and calcium sedimentation in aortic wall . Furthermore , foam cell formation induced by ox-LDL was decreased by hexarelin . In conclusion , hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in rats , possibly through upregulating HDL-c/LDL-c ratio , vascular NO production and downregulating the VSMC proliferation , aortic calcium sedimentation and foam cell formation . These novel anti-atherosclerotic actions of hexarelin suggest that the peptide might have a clinical potential in treating atherosclerosis . 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 . 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 . 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 . DB08860 inhibits vascular smooth muscle cell proliferation by inactivating extracellular signal-regulated kinases 1/2 . We recently reported that lysophosphatidylcholine ( lysoPC ) acts on vascular smooth muscle cells ( VSMCs ) to produce a mitogenic response through the activation of extracellular signal-regulated kinases 1/2 ( P27361 /2 ) . In this study , we examined the role of P04035 inhibitors on lysoPC-induced VSMC proliferation . DB08860 , a new P04035 inhibitor , suppressed lysoPC-induced DNA synthesis in primary cultured rat VSMCs . Since lysoPC-induced P27361 /2 activation contributes to smooth muscle cell proliferation , we explored the effect of pitavastatin on P27361 /2 activation . DB08860 inhibited lysoPC-induced P27361 /2 phosphorylation and P27361 /2 activation . The other P04035 inhibitors , atrovastatin and fluvastatin , also inhibited lysoPC-induced P27361 /2 phosphorylation . DB08860 also inhibited lysoPC-induced c-fos mRNA expression . To gain insight into the mechanism of the inhibitory effect of pitavastatin on P27361 /2 activation by lysoPC , we examined the role of the mevalonate pathways . Mevalonate and farnesylpyrophosphate reduced the inhibition of P27361 /2 phosphorylation by pitavastatin . These studies demonstrate that pitavastatin may inhibit lysoPC-induced VSMC proliferation , at least in part , by inactivating P27361 /2 , which is linked to mevalonate metabolism . DB00644 analogues reduce the proliferation of endometrial stromal cells but not endometriotic cells . AIMS : We investigated the potential of gonadotropin-releasing hormone ( DB00644 ) agonists and DB00644 antagonists to inhibit cell proliferation in endometriotic and endometrial stromal cells . METHODS : Twenty patients with ovarian endometriomas and 18 patients with uterine fibromas were recruited . Endometriotic and endometrial stromal cells were obtained from the ovarian chocolate cyst linings and the eutopic endometria of premenopausal women with uterine fibromas , respectively . RESULTS : DB00644 agonist or antagonist treatment attenuated tumor necrosis factor ( P01375 ) -α-induced cell proliferation in the endometrial stromal cells , whereas endometriotic stromal cells did not respond to treatment . The endometriotic stromal cells exhibited a decreased expression of the type I P30968 compared with the endometrial stromal cells . DB00644 agonists or antagonists did not repress P01375 -α-induced P10145 production in endometriotic stromal cells . CONCLUSION : DB00644 agonists and antagonists have similar effects in slowing the growth of endometrial stromal cells . Endometriotic stromal cells resist the antiproliferative effect of DB00644 agonists and antagonists . 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 . Proangiogenesis action of the thyroid hormone analog 3,5-diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2- to 3-fold by either T(4) or T(3) at 10(-8)-10(-7) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5-diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T(3) and T(4) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T(4) and T(3) for a novel cell surface hormone receptor site on integrin alphavbeta3 . The thyroid hormone analogs DITPA , T(4) , and T(4)-agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 /2 signal transduction cascade . Additionally , a specific integrin alphavbeta3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta3 , and are MAPK dependent . 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 . " Activated " P35610 proteins : a paradoxical consequence of inhibited JAK- P35610 signaling in cytomegalovirus-infected cells . We have previously characterized mouse CMV ( MCMV ) -encoded immune-evasive IFN signaling inhibition and identified the viral protein pM27 as inducer of proteasomal degradation of P52630 . Extending our analysis to P42224 and P40763 , we found that MCMV infection neither destabilizes P42224 protein nor prevents P42224 tyrosine Y701 phosphorylation , nuclear translocation , or the capability to bind γ-activated sequence DNA-enhancer elements . Unexpectedly , the analysis of P40763 revealed an induction of P40763 Y705 phosphorylation by MCMV . In parallel , we found decreasing P40763 protein amounts upon MCMV infection , although P40763 expression normally is positive autoregulative . P40763 phosphorylation depended on the duration of MCMV infection , the infectious dose , and MCMV gene expression but was independent of P17181 , P22301 , P05231 , and O60674 . Although P40763 phosphorylation did not require MCMV immediate early 1 , pM27 , and late gene expression , it was restricted to MCMV-infected cells and not transmitted to bystander cells . Despite intact P42224 Y701 phosphorylation , IFN-γ-induced target gene transcription ( e.g. , P10914 and suppressor of cytokine signaling [ Q9NSE2 ] 1 ) was strongly impaired . Likewise , the induction of P40763 target genes ( e.g. , O14543 ) by P05231 was also abolished , indicating that MCMV antagonizes P42224 and P40763 despite the occurrence of tyrosine phosphorylation . Consistent with the lack of O15524 induction , P42224 phosphorylation was prolonged upon IFN-γ treatment . We conclude that the inhibition of canonical P42224 and P40763 target gene expression abrogates their intrinsic negative feedback loops , leading to accumulation of phospho-tyrosine- P40763 and prolonged P42224 phosphorylation . These findings challenge the generalization of tyrosine-phosphorylated STATs necessarily being transcriptional active and document antagonistic effects of MCMV on P42224 /3-dependent target gene expression . 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 )

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Heterozygous Hb Hope [ beta136( O43583 ) DB00145 --> DB00128 ] in association with heterozygous beta0-thalassemia with apparent homozygous expression , in a Spanish patient . Hb Hope [ beta136( O43583 ) DB00145 --> DB00128 ( P19440 --> Q6IB77 ) ] has been found alone or in combination with other globin gene mutations in several African-American families , as well as in Japanese , Thai , Laotian , Cuban and Mauritanian families . We report the hematological and molecular characteristics of a heterozygous association of Hb Hope with beta0-thalassemia ( thal ) in a Spanish patient , in whom the level of expression of abnormal hemoglobin ( Hb ) by cation exchange high performance liquid chromatography ( HPLC ) and electrophoresis suggested initially a homozygous expression of the abnormal Hb , although sequencing of the polymerase chain reaction ( PCR ) -amplified beta-globin gene demonstrated a heterozygous genotype for Hb Hope . To the best of our knowledge , this is the first description of a case of Hb Hope in a Spanish family . Bacteroides fragilis enterotoxin induces cyclooxygenase-2 and fluid secretion in intestinal epithelial cells through NF-kappaB activation . Bacteroides fragilis produces an approximately 20-kDa heat-labile toxin ( B. fragilis enterotoxin , P78337 ) which is known to be associated with diarrhea . To determine whether cyclooxygenase ( P36551 ) -2 , via NF-kappaB activation , can contribute to P78337 -induced diarrhea , the relationship between P35354 expression and fluid secretion in P78337 -stimulated human intestinal epithelial cells was examined . P78337 stimulation increased the expression of P35354 , but not P23219 , in human intestinal epithelial cells . Suppression of the NF-kappaB signal significantly decreased P35354 expression in response to P78337 stimulation . DB00917 ( DB00917 ) levels were increased in parallel with P35354 expression , and , conversely , DB00917 production was significantly inhibited when P35354 or NF-kappaB activities were suppressed using P35354 small interfering RNA ( siRNA ) , p65 NF-kappaB subunit siRNA , or a retrovirus encoding the P25963 superrepressor . In addition , a selective P35354 inhibitor , NS-398 , significantly inhibited the increased DB02527 level induced by P78337 stimulation . Furthermore , a selective P35354 inhibitor prevented P78337 -induced DB00917 production and ileal fluid secretion in a mouse ileal loop model . These results suggest that the secretory response to P78337 stimulation may be mediated by the production of DB00917 , through NF-kappaB activation and the up-regulation of P35354 in intestinal epithelial cells . Comparison of rat and human parathyroid hormone 2 ( PTH2 ) receptor activation : PTH is a low potency partial agonist at the rat P49190 . The human P49190 , expressed in tissue culture cells , is selectively activated by PTH . Detailed investigation of its anatomical and cellular distribution has been performed in the rat . It is expressed by neurons in a number of brain nuclei ; by endocrine cells that include pancreatic islet somatostatin cells , thyroid parafollicular cells , and peptide secreting cells in the gastrointestinal tract ; and by cells in the vasculature and heart . The physiological role of the P49190 expressed by these cells remains to be determined . All pharmacological studies performed to date have used the human receptor . We have now isolated a complementary DNA including the entire coding sequence of the rat P49190 and compared its pharmacological profile with that of the human P49190 when each is expressed in COS-7 cells . PTH-based peptides , including rat DB05829 , rat PTH(1-34) , and human PTH(1-34) , have low potency at the rat P49190 for stimulation of adenylyl cyclase ( EC50 = 19-140 nM ) . When compared with the effect of a bovine hypothalamic extract , PTH-based peptides are partial agonists at the rat P49190 . This suggests that PTH is unlikely to be a physiologically important endogenous ligand for the P49190 . A peptide homologous to an activity detected in a bovine hypothalamic extract is a good candidate for the endogenous P49190 ligand . 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 . Effect of endothelin inhibition on lung fibroblasts on patients with systemic sclerosis . AIM : The aim of the study was to investigate the effect of selective P25101 DB06268 on viability and differentiation into myofibroblasts of lung fibroblasts derived from SSc-ILD patients and the ability of this drug to modify the lung fibroblast synthesis of P15692 , type I collagen and fibronectin . METHODS : Primary human lung fibroblast cultures were obtained from BAL of SSc-ILD patients . Cell cultures were exposed for 48 h to crescent concentrations of DB06268 ( 10 -6M to 10 -4M ) . In these experimental conditions we evaluated cell viability through crystal violet staining , the production and mRNA expression of P15692 , fibronectin and type I collagen respectively through ELISA and real-Time PCR . Further , we detected alpha-Smooth Muscle Actin ( α-SMA ) through immunocytochemical assay . RESULTS : The lowest concentration of sitaxsentan ( 10-6M ) did not affect fibroblasts viability ; conversely at higher concentrations , sitaxsentan induced a significant inhibition of cell viability . Synthesis and mRNA expression of P15692 , type 1 collagen and fibronectin were significantly reduced in treated lung fibroblasts compared to the untreated ones , in a dose-dependent manner . At higher concentrations , DB06268 reduced the expression of α-SMA . CONCLUSION : The results of this study show that sitaxentan is able in vitro to reduce both cell viability than production of P15692 and extra-cellular matrix components in SSc lung fibroblasts , confirming the anti-fibrotic potential of P25101 in SSc . The decreased expression of α-SMA in treated cells indicate that sitaxsentan may inhibit the fibroblast differentiation toward a myo-fibroblast phenotype and further support the hypothesis that the selective ETRAs may be beneficial in patients with SSc-ILD as anti fibrotic agents . Increased expression of cyclooxygenase-2 mediates enhanced contraction to endothelin P25101 receptor stimulation in endothelial nitric oxide synthase knockout mice . The aim of this study was to determine whether prolonged loss of NO activity , in endothelial NO synthase knockout ( P29474 (-/-) ) mice , influences endothelin ( ET ) P25101 receptor-mediated smooth muscle contraction and , if so , to define the underlying mechanism(s) . In isolated endothelium-denuded abdominal aortas , contractions to the selective P25101 receptor agonist ET-1(1-31) were significantly increased in aortas from P29474 (-/-) compared with wild-type ( WT ) mice . In contrast , contractions to the alpha1-adrenergic agonist phenylephrine or the thromboxane ( TX ) A2 analog U-46619 were similar between P29474 (-/-) and WT mice . Immunofluorescent and Western blot analysis demonstrated that the aortic expression of P25101 receptors was decreased in P29474 (-/-) compared with WT mice . Contractions evoked by ET-1(1-31) , but not phenylephrine , were reduced by inhibition of cyclooxygenase-2 ( P35354 ) ( indomethacin or celecoxib ) or of TXA2/prostaglandin H2 receptors ( SQ-29548 ) . After P36551 inhibition , contractions to ET-1(1-31) were no longer increased and were actually decreased in P29474 (-/-) compared with WT aortas . Western blot analysis revealed that endothelium-denuded abdominal aortas express P35354 , but not P23219 , and that expression of P35354 was significantly increased in P29474 (-/-) compared with WT mice . Contractions to the P36551 substrate arachidonic acid were also increased in P29474 (-/-) aortas . Furthermore , ET-1(1-31) but not phenylephrine stimulated production of the TXA2 metabolite TXB2 , which was increased in P29474 (-/-) compared with WT aortas . Therefore , P35354 plays a crucial and selective role in P25101 -mediated smooth muscle contraction . Furthermore , P35354 expression is increased in P29474 (-/-) mice , which overcomes a reduced expression of P25101 receptors and enables a selective increase in contraction to P25101 receptor stimulation . 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 . 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 . Somatostatin preserved blood brain barrier against cytokine induced alterations : possible role in multiple sclerosis . Multiple sclerosis ( MS ) is an inflammatory neurological disorder associated with demyelination , impaired blood brain barrier ( BBB ) , axonal damage and neuronal loss . In the present study , we measured somatostatin ( P61278 ) and tumor necrosis factor-α ( P01375 -α ) like immunoreactivity in P04141 samples from MS and non-MS patients . We also examined the role of P61278 in cytokines and lipopolysaccharide ( LPS ) -induced damage to the BBB using human brain endothelial cells in culture . Most of the cerebrospinal fluid ( P04141 ) samples studied from definite MS patients exhibited lower somatostatin ( P61278 ) -like immunoreactivity and higher expression of P01375 -α in comparison to non-MS patients . Treatment of cells with cytokines and LPS blocked P61278 secretion and decreased P61278 expression . Human brain endothelial cells expressed all five somatostatin receptors ( SSTRs ) with increased expression of P30874 and 4 upon treatment with cytokines and LPS . Cytokines and LPS-induced disruption of the tight junction proteins Zonula occludens ( ZO-1 ) organization was restored in presence of P61278 , P30874 or P31391 selective agonists . Furthermore , inflammation induced changes in extracellular signal-regulated kinases ( P27361 /2 and Q13164 ) signaling and altered expression of endothelial and inducible nitric oxide synthase are modulated in presence of P61278 . These data indicate that decreased levels of P61278 contribute to failure of the BBB in MS . P04626 -directed therapy for metastatic breast cancer . Human epidermal growth factor receptor 2 ( P04626 ) overexpression drives the biology of 20 % of breast cancers , and predicts a poor prognosis for patients . P04626 -targeted therapies significantly improve outcomes for P04626 -positive patients with both early and metastatic breast cancer . Currently three P04626 -targeted agents , trastuzumab ( Herceptin ) , lapatinib ( DB01259 ) , and pertuzumab ( Perjeta ) , are available for the treatment of P04626 -positive metastatic breast cancer ( MBC ) . Numerous studies have attempted to optimize their use by combining them with each other , or with endocrine and cytotoxic therapies . Most recently , the FDA approved the combination of trastuzumab , pertuzumab , and docetaxel as first-line treatment for MBC , and in late February 2013 approved a fourth P04626 -targeted agent , trastuzumab emtansine ( DB05773 , Kadcyla ) , for accelerated approval . These advances create a number of clinical dilemmas , including identification of the optimal sequence of P04626 -targeted agents and the best drug combinations to use , as well as the recognition of primary and acquired drug resistance . In this article , we review clinical data informing the effective management of P04626 -positive MBC . Effects of Subchronic Treatment with Ibuprofen and DB04743 on Spatial Memory and NMDAR Subunits Expression in Aged Rats . Several studies point to an important function of cyclooxygenase ( P36551 ) and prostaglandin signaling in models of synaptic plasticity which is associated with N-methyl-D-aspartate receptors ( NMDARs ) . Cyclooxygenase gene is suggested to be an immediate early gene that is tightly regulated in neurons by DB01221 dependent synaptic activity . Nonsteroid Antiinflammatory Drugs ( NSAIDs ) exert their antiinflammatory effect by the inhibion of P36551 have controversial effects on learning and memory . We administered ibuprofen as a non-selective P35354 inhibitor and nimesulide as a selective P35354 inhibitor for 8 weeks for determining the cognitive impact of subchronic administration of NSAIDs to aged rats . Wistar albino rats ( 16 mo , n = 30 ) were separated into control ( n = 10 ) , ibuprofen ( n = 10 ) and nimesulide ( n = 10 ) treated groups . First we evaluated hippocampus-dependent spatial memory in the radial arm maze ( RAM ) and than we evaluated the expression of the NMDAR subunits , Q12879 and Q13224 by western blotting to see if their expressions are effected by subchronic administration with these drugs . Ibuprofen and nimesulide treated rats completed the task in a statistically significant shorter time when compared with control group ( p < 0.01 ) , but there was no statistically significant difference between groups about choice accuracy data in RAM . Furthermore , no statistically significant difference was detected for the protein expressions of Q12879 and Q13224 of the subjects . Oral administration of ibuprofen and nimesulide for 8 weeks showed no impairment but partly improved spatial memory . Novel synthesis of various orthogonally protected Cα-methyllysine analogues and biological evaluation of a vapreotide analogue containing ( S ) -α-methyllysine . Prochiral malonic diesters containing a quaternary carbon center have been successfully transformed into a diverse set of (t)Boc-Fmoc-α(2,2)-methyllysine-OH analogues through chiral malonic half-ester intermediates obtained via enzymatic ( Pig Liver Esterase , PLE ) hydrolysis . The variety of chiral half-ester intermediates , which vary from 1 to 6 methylene units in the side chain , are achieved in moderate to high optical purity and in good yields . The PLE hydrolysis of malonic diesters with various side chain lengths appears to obey the Jones 's PLE model according to the stereochemical configurations of the resulting chiral half-esters . The established synthetic strategy allows the construction of both enantiomers of α(2,2)-methyllysine analogues , and a ( S ) -β(2,2)-methyllysine analogue from a common synthon by straightforward manipulation of protecting groups . Two different straightforward and cost effective synthetic strategies are described for the synthesis of α(2,2)-methyllysine analogues . The described strategies should find significant usefulness in preparing novel peptide libraries with unnatural lysine analogues . A DB04894 analogue incorporating ( S ) -α(2,2)-methyllysine was prepared . However , the DB04894 analogue with ( S ) -α-methyl-α-lysine is found to lose its specific binding to somatostatin receptor subtype 2 ( P30874 ) . Celecoxib inhibits serum amyloid a-induced matrix metalloproteinase-10 expression in human endothelial cells . BACKGROUND : Although serum amyloid A ( P0DJI8 ) is an established biomarker of coronary artery disease ( CAD ) , its direct role in matrix degradation is obscure . This study investigated the effect of P0DJI8 on the expression of matrix metalloproteinase-10 ( P09238 ) in endothelial cells . The effect of celecoxib on P0DJI8 -dependent P09238 expression and its possible mediator were also investigated . METHODS AND RESULTS : From our time course microarray screening , P0DJI8 ( 20 microg/ml ) was found to increase P09238 mRNA expression over time ( 4-48 h ) in human endothelial cells . Quantitative real-time PCR confirmed this transcriptional induction . Correspondingly , secreted P09238 protein was also markedly induced by P0DJI8 treatment for 24 h in a dose-dependent manner . We further examined cyclooxygenase-2 ( P35354 ) and its major product , prostaglandin E(2) ( PGE(2) ) , as possible mediators of P09238 induction . Direct PGE(2) treatment could result in P09238 induction . Celecoxib , a selective P35354 inhibitor , suppressed P09238 secretion induced by P0DJI8 . CONCLUSIONS : P0DJI8 induced P09238 expression and celecoxib prevented its induction . P09238 induction was at least partly mediated by PGE(2) . P49190 -mediated inhibitory effect of parathyroid hormone and Q96A98 on cell proliferation . The parathyroid hormone ( PTH ) has dual mitogenic and inhibitory effects on cell proliferation , depending on the cell type and experimental conditions . PTH can signal via two different receptors , both positively coupled to the adenylyl cyclase/cyclic AMP pathway which can mimic some of the proliferative effects of PTH . We evaluated the role of the type-2 PTH ( PTH2 ) receptor on cell proliferation in clonal human embryonic kidney HEK293 cells stably expressing the human P49190 . Using a cyclic AMP-responsive gene-reporter , we confirmed that the tuberoinfundibular peptide ( Q96A98 ) and various human ( h ) PTH fragments including DB05829 -(1-34) were potent agonists ( EC(50) in the range of 0.01-0.3 nM ) whereas the bovine ( b ) PTH peptides b( DB00135 (34))PTH-(7-34) and its tryptophan derivative b[D- DB00150 (12), DB00135 (34)]PTH-(7-34) behaved as antagonists ( IC(50)=117 and 249 nM , respectively ) . DB05829 -(1-34) produced a dose-dependent inhibition of cell proliferation ( EC(50)=8.5+/-0.4 nM ) after 3 days and this effect was fully reversed by the tryptophan derivative antagonist . The same effect was observed with Q96A98 which caused a 30 % maximal inhibition . These findings reveal that P49190 activation can inhibit cell proliferation and might explain the dual functionality of PTH on cell proliferation . Case report : DB00065 treatment in two Chinese patients with psoriatic arthritis . Psoriatic arthritis ( PsA ) is a rheumatoid factor ( RF ) -seronegative systemic inflammatory disorder associated with psoriasis . Current treatment for PsA in China is still focused on disease modifying anti-rheumatic drugs ( DMARDs ) . In this paper , we report two Chinese patients with active longstanding PsA treated with infliximab , a human-mouse chimeric monoclonal antibody against tumor necrosis factor alpha ( P01375 -α ) . The results show that infliximab acted quickly and effectively in relieving peripheral and axial symptoms and refractory skin lesions , even in recombinant human P01375 -α receptor ( rhTNFR ) -resistant case . The take-home message from our cases is that infliximab is a useful therapeutic option for refractory PsA , especially when a patient has a combination of psoriasis and psoriatic arthritis . Further local evidence and experience must be accumulated in order to make anti- P01375 -α therapy more accessible to PsA patients in China . Effect of Electroacupuncture Intervention on Expression of P80511 , SP , P23219 , and DB00917 of Dorsal Portion of the Cervical Spinal Cord in Rats with Neck-Incision Pain . The present study was aimed to determine if cervicospinal DB05875 ( SP ) and its neurokinin-1 receptor ( P25103 ) , calcitonin gene-related peptide ( P80511 ) , cyclooxygenase-1 ( P23219 ) , and prostaglandin E2 ( DB00917 ) were involved in electroacupuncture ( EA ) analgesia in neck-incision pain rats . EA intervention was applied to bilateral Futu ( LI18 ) , Hegu ( LI4 ) -Neiguan ( Q92824 ) , and Zusanli ( ST36 ) -Yanglingquan ( GB34 ) for 30 min . Cervicospinal SP and P80511 immunoactivity was detected by immunofluorescence technique , P25103 and P23219 protein and mRNA expression levels were determined using Western blot and real-time PCR , respectively , and DB00917 content was measured using ELISA . Outcomes indicated that EA of EA-LI18 and LI4- Q92824 ( not ST36-GB34 ) significantly suppressed neck-incision induced decrease of thermal pain threshold ( P < 0.05 ) . EA stimulation of LI18 and LI4- Q92824 markedly inhibited neck-incision induced upregulation of SP and P80511 immunoactivity , NK-1 R and P23219 mRNA and protein expression levels , as well as the increase of DB00917 content in the dorsal cervicospinal cord ( P < 0.05 ) . These findings showed that LI18 and LI4- Q92824 EA stimulation-induced downregulation of SP , P80511 , P25103 , P23219 , and DB00917 levels in the dorsal cervicospinal cord may contribute to their effects in relieving neck-incision pain . This study highlights the targets of EA intervention for reducing post-thyroid-surgery pain for the first time . Analysis of serum amyloid A1 exon 4 polymorphism in Japanese population . Nucleotide variation in the triplet codon coding for amino acid position 72 of human serum amyloid A1 ( P0DJI8 ) , which was suggested by amino acid sequence analysis , was analyzed here in order to identify the genomic sequences coding P0DJI8 .2 and to characterize further the P0DJI8 allele frequency in a Japanese population . The P0DJI8 exon 4 was amplified by PCR and treated with Nco I . Sequencing of PCR products from genomic DNA of individuals who were heterozygous for the Nco I site revealed P19440 ( DB00145 ) and Q6IB77 ( DB00128 ) at the position 72 . The allele having 72Asp showed the exon 3 polymorphism coding 52Ala and 57Val . This allele should thus be identified as P0DJI8 .2 . Alleles with 72Gly were either 52Val and 57Ala( P0DJI8 .1) or 52Ala and 57Ala ( P0DJI8 .3 ) or 52Ala and 57Val ( P0DJI8 .5 ) . The frequency of P0DJI8 alleles in the 321 Japanese subjects was 0.310 , 0.012 , 0.347 and 0.330 for each P0DJI8 allel of 1.1 , 1.2 , 1.3 and 1.5 , respectively . The presence of the P0DJI8 .4 allele was not evaluated . Combination of sitaxentan and tadalafil for idiopathic pulmonary arterial hypertension following relapse on DB00559 . DB06268 , a highly-selective endothelin receptor antagonist ( P25101 ) and DB00559 a non-selective P25101 are both approved for the treatment of idiopathic pulmonary arterial hypertension ( iPAH ) . DB00203 is a phosphodiesterase-5 ( PDE-5 ) inhibitor used in the treatment of iPAH . DB00820 is a long acting PDE-5 inhibitor largely unexplored for the treatment of iPAH . Following failure of monotherapy combination therapy with an P25101 and a PDE-5 inhibitor is often used , a frequently used combination being DB00559 with sildenafil . We report our clinical experience in three patients with iPAH treated with a combination of sitaxentan and tadalafil , who previously discontinued DB00559 . There was sustained symptomatic and haemodynamic improvement in all three patients treated with the combination . No adverse effect related to the combination treatment was noted . DB06268 and tadalafil , both being once a day treatments , can also possibly increase compliance . 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 . 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 . Molecular events caused by mechanical stress in bone . The shape of bone changes as a result of bone remodeling corresponding to physical circumstances such as mechanical stress . The tissue which receives the loaded mechanical stress most efficiently is bone matrix . Recent studies revealed the function of osteocytes as mechanosensors in the early stage of bone remodeling . Loaded mechanical stress is converted to a series of biochemical reactions , and finally activates osteoclasts and osteoblasts to cause bone resorption and formation . Biochemical and molecular biological studies have recently resulted in the identification of the gene of which expression level is changed by mechanical stress . DB00435 ( NO ) and DB02527 is secreted in response to mechanical stress in the immediate early stage . Genes encoding enzymes such as glutamate/aspartate transporter ( P43003 ) , nitric oxide synthetase ( NOS ) and prostaglandin G/H synthetase ( P35354 ) are identified as mechanical stress-responsive . The expression level of P05019 is enhanced under the control of PTH/ P12272 . The expression of c-fos is increased by loading of mechanical stress . P05412 , a heterodimer of c- P01100 /c- P05412 , functions as a transcription factor of downstream gene(s) . Elements including P05412 sites , cyclic AMP response elements ( CRE ) and shear stress response elements ( SSRE ) are found in the promoter region of mechanical stress-response genes . The enhanced expression of osteopontin ( P10451 ) in the osteocytes of bone resorption sites was demonstrated by in situ hybridization and immunohistochemistry and transdifferentiation of chondrocytes with the abundant expression of P12643 and -4 in the process of distraction osteogenesis was observed . Interactions among positions in the third and fourth membrane-associated domains at the intersubunit interface of the N-methyl-D-aspartate receptor forming sites of alcohol action . The N-methyl-D-aspartate ( DB01221 ) glutamate receptor is a major target of ethanol in the brain . Previous studies have identified positions in the third and fourth membrane-associated ( M ) domains of the DB01221 receptor Q05586 and Q12879 subunits that influence alcohol sensitivity . The predicted structure of the DB01221 receptor , based on that of the related P42262 subunit , indicates a close apposition of the alcohol-sensitive positions in M3 and M4 between the two subunit types . We tested the hypothesis that these positions interact to regulate receptor kinetics and ethanol sensitivity by using dual substitution mutants . In single-substitution mutants , we found that a position in both subunits adjacent to one previously identified , Q05586 ( DB00145 -638) and Q12879 ( DB00120 -636) , can strongly regulate ethanol sensitivity . Significant interactions affecting ethanol inhibition and receptor deactivation were observed at four pairs of positions in Q05586 / Q12879 : DB00145 -638/ DB00134 -823 , DB00120 -639/ DB00149 -824 , DB00134 -818/ DB00120 -636 , and DB00149 -819/ DB00120 -637 ; the latter pair also interacted with respect to desensitization . Two interactions involved a position in M4 of both subunits , Q05586 ( DB00134 -818) and Q12879 ( DB00149 -824) , that does not by itself alter ethanol sensitivity , whereas a previously identified ethanol-sensitive position , Q12879 (Ala-825) , did not unequivocally interact with any other position tested . These results also indicate a shift by one position of the predicted alignment of the Q05586 M4 domain . These findings have allowed for the refinement of the DB01221 receptor M domain structure , demonstrate that this region can influence apparent agonist affinity , and support the existence of four sites of alcohol action on the DB01221 receptor , each consisting of five amino acids at the M3-M4 domain intersubunit interfaces . 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 . Zebrafish express the common parathyroid hormone/parathyroid hormone-related peptide receptor ( Q03431 ) and a novel receptor ( PTH3R ) that is preferentially activated by mammalian and fugufish parathyroid hormone-related peptide . To further explore the evolution of receptors for parathyroid hormone ( PTH ) and PTH-related peptide ( P12272 ) , we searched for zebrafish ( z ) homologs of the PTH/ P12272 receptor ( Q03431 ) . In mammalian genes encoding this receptor , exons M6/7 and M7 are highly conserved and separated by 81-84 intronic nucleotides . Genomic polymerase chain reaction using degenerate primers based on these exons led to two distinct DNA fragments comprising portions of genes encoding the zPTH1R and the novel zPTH3R . Sequence comparison of both full-length teleost receptors revealed 69 % similarity ( 61 % identity ) , but less homology with zPTH2R . When compared with hPTH1R , zPTH1R showed 76 % and zPTH3R 67 % amino acid sequence similarity ; similarity with hPTH2R was only 59 % for both teleost receptors . When expressed in COS-7 cells , zPTH1R bound [ DB00135 (34)] DB05829 -(1-34)-amide ( DB05829 ) , [ DB00135 (36)]hPTHrP-(1-36)-amide ( hPTHrP ) , and [ Ala(29), DB00142 (30) , Ala(34), DB00142 (35) , DB00135 (36) ] fugufish P12272 -(1-36)-amide ( fuguPTHrP ) with a high apparent affinity ( IC(50) : 1.2-3.5 nM ) , and was efficiently activated by all three peptides ( EC(50) : 1.1-1.7 nM ) . In contrast , zPTH3R showed higher affinity for fuguPTHrP and hPTHrP ( IC(50) : 2.1-11.1 nM ) than for DB05829 ( IC(50) : 118.2-127.0 nM ) ; DB02527 accumulation was more efficiently stimulated by fugufish and human P12272 ( EC(50) : 0.47 +/- 0.27 and 0.45 +/- 0.16 , respectively ) than by DB05829 ( EC(50) : 9.95 +/- 1.5 nM ) . Agonist-stimulated total inositol phosphate accumulation was observed with zPTH1R , but not zPTH3R . Common single nucleotide polymorphisms in cyclooxygenase-2 and risk of severe chronic periodontitis in a Chinese population . AIM : Several common single nucleotide polymorphisms ( SNPs ) of the cyclooxygenase-2 ( P35354 ) gene have been reported to be functional . The association between -1195GA , -765GC and 8473TC of P35354 , and severe chronic periodontitis ( CP ) in a Chinese population was investigated . MATERIAL AND METHODS : 148 cases of healthy controls ( control group ) and 146 cases of severe CP were recruited in this study . Genotypes of -1195GA , -765GC and 8473TC were determined by polymerase chain reaction restriction fragment length polymorphism ( PCR-RFLP ) . The distributions of genotypes and haplotypes were compared by chi(2) test and the odds ratios ( ORs ) were calculated by logistic regression analysis . RESULTS : The prevalence of the -1195A was more prevalent in CP group ( 60.62 % ) than control group ( 51.35 % ) , and the distributions of the -765C and 8473C were higher in control group ( 6.76 % and 21.96 % ) compared with CP group ( 3.08 % and 15.07 % ) . Only genotype distribution of -1195GA was significant when p-value was corrected for multiple testing ( p(c)=0.033 ) . The adjusted ORs for the -1195AA/GA , -765GC and 8473CC/TC were 2.49 ( 95 % CI=1.33-4.69 , p=0.005 ) , 0.45 ( 95 % CI=0.20-1.04 , p=0.061 ) and 0.67 ( 95 % CI=0.41-1.11 , p=0.118 ) . Subjects with the haplotype AGT had a significantly higher risk of periodontitis than those with the most common haplotype P19440 ( OR=1.91 , 95 % CI=1.32-2.76 , p(c) < 0.001 ) . CONCLUSIONS : It suggests the -1195A variant is associated with an increased risk for severe CP . DB00072 emtansine in breast cancer . INTRODUCTION : DB00072 emtansine ( DB05773 ) is a human epidermal growth factor receptor 2 ( P04626 ) -targeted antibody-drug conjugate ( ADC ) composed of trastuzumab , a stable linker ( MCC ) , and the cytotoxic agent DM1 ( derivative of maytansine ) . Administration of DB05773 leads to limited systemic exposure of free DM1 , with no evidence of DM1 accumulation after repeated dosing . AREAS COVERED : Phase I and Phase II clinical trials with DB05773 as a single agent and in combination with paclitaxel , docetaxel , and pertuzumab have shown substantial clinical activity and a favorable safety profile . A randomized , open-label , first-line trial comparing trastuzumab and docetaxel with single agent DB05773 showed a significant improved progression-free survival for DB05773 . EXPERT OPINION : DB05773 has successfully completed second-line Phase III development for advanced P04626 -positive breast cancer . The Phase III EMILIA study demonstrated an overall survival benefit for DB05773 compared to the combination of lapatinib and capecitabine in taxane-trastuzumab pretreated patients . DB05773 may offer delivery on a personalized basis of very potent cytotoxic agents in a cellular selective manner . Exendin-4 attenuates lipopolysaccharides induced inflammatory response but does not protects H9c2 cells from apoptosis . BACKGROUND : Glucagon-like peptide-1 ( P0C6A0 ) and its analogues are reported to exert wide-ranging cardiovascular actions in preclinical and clinical studies . We thus investigated whether the P43220 agonist , exendin-4 , has inhibitory effects on LPS-stimulated inflammatory response in cardiomyoblasts . METHODS : H9c2 cardiomyoblasts were exposed to LPS and treated with exendin-4 . Expressions of proinflammatory mediators were assessed using quantitative real-time PCR . Nuclear localization of NF-κB was examined using immunoblotting . mRNA expression of inducible nitric oxide synthase ( P35228 ) and nitric oxide ( NO ) production were evaluated by q PCR and NO assay . Furthermore , anti-apoptotic effect of exendin-4 in LPS-stimulated H9c2 cells was determined using qPCR and immunoblot . RESULTS : Exposure to LPS increased mRNA expressions of P01375 -α , P35354 and P14780 in H9c2 cells . It also caused increases in P35228 mRNA expression and NF-κB nuclear translocation . Exendin-4 dose-dependently downregulated mRNA levels of P01375 -α , P35354 and P14780 in LPS-stimulated H9c2 cells . It also reduced NF-κB nuclear translocation . Treatment with exendin-4 showed no effect on LPS-induced apoptosis in H9c2 cells . CONCLUSIONS : Exendin-4 exerts an effect on cardiomyoblast exposed to LPS by inhibiting mRNA expression of inflammatory mediators and suppressing NF-κB activation . These effects are consistent with some of the observed anti-inflammatory properties of exendin-4 , as well as its beneficial actions on the cardiovascular system . DB00065 administered with shortened infusion times in a specialized Q9UKU7 infusion unit : a prospective cohort study . BACKGROUND AND AIMS : Biological therapy with anti P01375 agents requires parenteral administration and in the case of infliximab this involves in hospital treatment . We aimed to prospectively assess the safety and tolerance of infliximab infusion in patients with Q9UKU7 in a specialized unit adhering to strict standard operation procedures including switch to accelerated 1h infusions . METHODS : A prospective audit of a referral center Q9UKU7 infusion unit was performed . We recorded infusion times and all adverse events including hypersensitivity reactions . Patients were also polled about the impact of the treatment on quality of life ( QOL ) . RESULTS : On 20 consecutive days 177 patients were treated with infliximab and all participated . Of those infliximab 117 received 1h infusions and 4 ( 2.2 % ) had an immediate infusion reaction . Median time on unit was optimal for those with 1h infusions [ 1:35 h ( IQR : 1:25-1:50 ) ] without an increased risk of infusion reactions . Prophylactic therapy significantly increased the time on unit [ 3:20 h ( IQR : 2:50-3:45 ) , p < 0.001 ] . Patients reported a high global satisfaction and a good tolerability of the infusions with a considerable or strong impact on studies , work or QOL in one third . CONCLUSIONS : A dedicated Q9UKU7 infusion unit can achieve high quality of care and shortened 1h infliximab infusions are well tolerated in patients with scheduled maintenance therapy . Analog of somatostatin vapreotide exhibits biological effects in vitro via interaction with neurokinin-1 receptor . OBJECTIVES : DB04894 , a synthetic analog of somatostatin , has analgesic activity most likely mediated through the blockade of neurokinin-1 receptor ( P25103 ) , the DB05875 ( SP ) -preferring receptor . The ability of vapreotide to interfere with other biological effects of SP has yet to be investigated . METHODS : We studied the ability of vapreotide to antagonize P25103 in three different cell types : immortalized U373MG human astrocytoma cells , human monocyte-derived macrophages ( MDM ) and a human embryonic kidney cell line , HEK293 . Both U373MG and MDM express endogenous P25103 while HEK293 cells , which normally do not express P25103 , are stably transformed to express human P25103 ( HEK293- P25103 ) . RESULTS : DB04894 attenuates SP-triggered intracellular calcium increases and nuclear factor-κB activation in a dose-dependent manner . DB04894 also inhibits SP-induced interleukin-8 and monocyte chemotactic protein-1 production in HEK293- P25103 and U373MG cell lines . DB04894 inhibits HIV-1 infection of human MDM in vitro , an effect that is reversible by SP pretreatment . CONCLUSIONS : Our findings indicate that vapreotide has P25103 antagonist activity and may have a potential application as a therapeutic intervention in HIV-1 infection . DB09045 : the newest P43220 agonist for the management of type 2 diabetes . OBJECTIVE : To review the pharmacology , pharmacokinetics , safety , and efficacy of the glucagon-like peptide-1 receptor agonist ( P0C6A0 RA ) , dulaglutide , in the treatment of type 2 diabetes mellitus ( T2D ) . DATA SOURCES : A PubMed search was completed to identify publications from 1947 to October 2014 using the search terms dulaglutide and LY2189265 . References were reviewed to identify additional resources . STUDY SELECTION AND DATA EXTRACTION : Articles were included if they evaluated the pharmacology , pharmacokinetics , safety , or efficacy of dulaglutide . DATA SYNTHESIS : DB09045 reduces both glycosylated hemoglobin ( A1C ) and weight by stimulating insulin secretion and suppressing glucagon in a glucose-dependent manner , delaying gastric emptying , and promoting satiety . DB09045 consists of 2 P0C6A0 analogues that have been modified to make it a long-acting , once-weekly agent . DB09045 has been studied as monotherapy and in combination with metformin , glimepiride , pioglitazone , and insulin lispro . It has demonstrated superior A1C reduction compared with placebo , metformin , insulin glargine , sitagliptin , and twice-daily exenatide . It demonstrated noninferiority in A1C reduction to liraglutide . DB09045 changed A1C by -0.78 % to -1.51 % , and it changed weight by -0.35 kg to -3.03 kg . The most common adverse effects in clinical studies were nausea , vomiting , and diarrhea . CONCLUSIONS : DB09045 is the fifth P0C6A0 RA approved for T2D in the United States . It is an attractive option because it is dosed once-weekly , provides A1C lowering similar to liraglutide , weight reduction similar to exenatide , and has an adverse effect profile similar to exenatide and liraglutide . Pharmacokinetic profiles of the novel P35354 selective inhibitor cimicoxib in dogs . DB05095 ( CX ) is a novel imidazole derivative that is a cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drug and the latest P35354 selective inhibitor to be released for veterinary use . Currently there is limited information available on the pharmacokinetic ( PK ) properties of CX . The aim of the current study was to evaluate the PK features of CX after administration of the recommended dose and after administration of a more variable dose rate in the form of the commercially available tablet . In addition , the effects of food intake on the PK properties were also evaluated . In the first study , five healthy Beagle dogs received 2mg/kg CX via the oral route following a period of fasting . The second study was conducted using six healthy Labrador retriever dogs which each received an 80 mg tablet ( approximate dose 1.95-2.5mg/kg ) using a crossover design , both in the fasted and fed condition . The plasma concentrations of CX were detected by a validated HPLC method . No adverse effects were observed in any dogs during the experiment . The results from the PK analysis were similar between the studies , regardless of precision of dose and fasted and fed conditions . The mean peak concentration of CX was 0.49 and 0.43 μg/mL under fasted and fed conditions , respectively . The mean half-life was about 3h after all treatments . In addition , simulated multiple dosing data revealed that time over minimal effective concentration was similar after 1.95 , 2.0 and 2.5mg/kg dose administrations . These findings suggest that slight variation from the recommended dose should not alter the therapeutic outcome . In addition , CX can be administered to fed dogs without significantly affecting blood levels . Q96A98 ( Q96A98 ) signaling modulates acute and tonic nociception . Q96A98 ( Q96A98 ) synthesizing neurons at the caudal border of the thalamus and in the lateral pons project to areas rich in its receptor , the parathyroid hormone 2 receptor ( P49190 ) . These areas include many involved in processing nociceptive information . Here we examined the potential role of Q96A98 signaling in nociception using a P49190 antagonist ( HYWH ) and mice with deletion of Q96A98 's coding sequence or P49190 null mutation . Intracerebroventricular ( icv ) infusion of HYWH significantly inhibited nociceptive responses in tail-flick and hot-plate tests and attenuated the nociceptive response to hindpaw formalin injection . Q96A98 -KO and P49190 -KO had increased response latency in the 55°C hot-plate test and reduced responses in the hindpaw formalin test . The tail-flick test was not affected in either KO line . Thermal hypoalgesia in KO mice was dose-dependently reversed by systemic administration of the cannabinoid receptor 1 ( P21554 ) antagonist rimonabant , which did not affect nociception in wild-type ( WT ) . Systemic administration of the cannabinoid agonist CP 55,940 did not affect nociception in KO mice at a dose effective in WT . WT mice administered HYWH icv , and both KOs , had significantly increased stress-induced analgesia ( SIA ) . DB06155 blocked the increased SIA in Q96A98 -KO , P49190 -KO or after HYWH infusion . P21554 and FAAH mRNA were decreased and increased , respectively , in the basolateral amygdala of Q96A98 -KO mice . These data suggest that Q96A98 signaling modulates nociception , very likely by inhibiting endocannabinoid circuitry at a supraspinal level . We infer a new central mechanism for endocannabinoid regulation , via Q96A98 acting on the P49190 in discrete brain regions . 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 . Antineoplastic mechanisms of niclosamide in acute myelogenous leukemia stem cells : inactivation of the NF-kappaB pathway and generation of reactive oxygen species . NF-kappaB may be a potential therapeutic target for acute myelogenous leukemia ( AML ) because NF-kappaB activation is found in primitive human AML blast cells . In this report , we initially discovered that the potent antineoplastic effect of niclosamide , a Food and Drug Administration-approved antihelminthic agent , was through inhibition of the NF-kappaB pathway in AML cells . DB06803 inhibited the transcription and DNA binding of NF-kappaB . It blocked tumor necrosis factor-induced P25963 phosphorylation , translocation of p65 , and expression of NF-kappaB-regulated genes . DB06803 inhibited the steps TAK1 --> O15111 ( IKK ) and IKK --> P25963 . DB06803 also increased the levels of reactive oxygen species ( ROS ) in AML cells . Quenching ROS by the glutathione precursor DB06151 attenuated niclosamide-induced apoptosis . Our results together suggest that niclosamide inhibited the NF-kappaB pathway and increased ROS levels to induce apoptosis in AML cells . On translational study of the efficacy of niclosamide against AML , niclosamide killed progenitor/stem cells from AML patients but spared those from normal bone marrow . DB06803 was synergistic with the frontline chemotherapeutic agents cytarabine , etoposide , and daunorubicin . It potently inhibited the growth of AML cells in vitro and in nude mice . Our results support further investigation of niclosamide in clinical trials of AML patients . Activation of transcription factor Nrf-2 and its downstream targets in response to moloney murine leukemia virus ts1-induced thiol depletion and oxidative stress in astrocytes . The neuroimmunodegenerative syndrome that develops in mice infected with ts1 , a mutant of Moloney murine leukemia virus , resembles human AIDS . Both ts1 and human immunodeficiency virus type 1 infect astrocytes , microglia , and oligodendrocytes but do not infect neurons . Oxidative stress has been implicated in the neuropathology of AIDS dementia and other neurodegenerative diseases . We report here that ts1 infection of astrocytes ( both transformed C1 cells and primary cultures ) also induces thiol ( i.e. , glutathione and cysteine ) depletion and reactive oxygen species ( ROS ) accumulation , events occurring in parallel with viral envelope precursor gPr80(env) accumulation and upregulated expression of endoplasmic reticulum chaperones P11021 and P14625 . Furthermore , ts1-infected astrocytes mobilize their thiol redox defenses by upregulating levels of the Nrf-2 transcription factor , as well its targets , the Q9UPY5 cystine/glutamate antiporter , gamma-glutamylcysteine ligase , and glutathione peroxidase . Depleting intracellular thiols by treating uninfected astrocytes with buthionine sulfoximine ( BSO ) , a glutathione synthesis inhibitor , or by culturing in cystine-deficient medium , also induces ROS accumulation , activates Nrf-2 , and upregulates Nrf-2 target gene expression in these astrocytes . Overexpression of Nrf-2 in astrocytes specifically increases expression of the above thiol synthesis-related proteins . Further treatment with BSO or DB06151 in transfected cells modulates this expression . Thiol depletion also accelerates cell death , while thiol supplementation promotes survival of ts1-infected cells . Together , our results indicate that ts1 infection of astrocytes , along with ts1-induced gPr80(env) accumulation , endoplasmic reticulum stress , thiol depletion , and oxidative stress , accelerates cell death ; in response to the thiol depletion and oxidative stress , astrocytes activate their Nrf-2-mediated thiol antioxidant defenses , promoting cell survival . 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 . Detection and quantification of cimicoxib , a novel P35354 inhibitor , in canine plasma by HPLC with spectrofluorimetric detection : development and validation of a new methodology . DB05095 ( CX ) is a selective P35354 inhibitor recently launched on the veterinary market . No analytical method to detect CX in biological samples has been published to date . The chromatographic separation was performed with a Kinetex C18 analytical column ( 100 mm × 4.6 mm , 2.6 μm particle size ) at 25 °C . The mobile phase consisted of acetonitrile:buffer ( 10 mM AcONH4 , pH 4.5 ) ( 35:65 , v/v ) at a flow rate of 1 mL/min . Excitation and emission wavelengths were 268 and 430 nm , respectively . The extraction used 500 μL of plasma added to 100 μL of IS ( 5 μg/mL ) and 100 μL of 10 % CF3COOH , extracted with 600 μL of C2H2:Et2O ( 3:7 , v/v ) . The organic phase was evaporated and reconstituted with 200 μL of mobile phase . The CX recovery ranged from 74.5 % to 82.6 % . The limit of quantification was 25 ng mL(-1) . The chromatographic runs were specific with no interfering peaks at the retention times of the analytes , as confirmed by HPLC-mass spectrometry experiments . The other validation parameters were in agreement with the international guidelines . The method was successfully tested on two dogs treated at two dose rates . It facilitated tracking of the plasma concentration for 24h and calculation of the main pharmacokinetic parameters . In conclusion , this method ( extraction , separation and applied techniques ) is simple , effective and specific . This is the first time that a method for the quantification of CX in plasma has been reported . This technique may have applications for further pharmacokinetic studies . DNA-binding activity of NF-kappaB and phosphorylation of p65 are induced by DB06151 through phosphatidylinositol ( PI ) 3-kinase . N-Acetylcysteine ( Q9C000 ) has been widely used as an antioxidant in research , however , it has also been found to reduce the binding of P01375 to its receptor independent of its antioxidative role . In this study , we investigated the effect of Q9C000 on NF-kappaB activation . In HeLa cells , Hep3B cells , and A549 cells , DNA-binding activity of NF-kappaB was induced by Q9C000 without any other stimulation but not by tetramethylthiourea ( TMTU ) or vitamin C , suggesting that ROS is not involved in the effect of Q9C000 . The degradation of P25963 and nuclear translocation of NF-kappaB were not induced by Q9C000 . The phosphorylation of p65 at serine 536 was induced by Q9C000 , which is known to contribute to the enhancement of DNA-binding activity of NF-kappaB , however , Q9C000 did not directly phosphorylate p65 . The Q9C000 -induced DNA-binding activity of NF-kappaB and phosphorylation of p65 were sensitive to a phosphatidylinositol ( PI ) 3-kinase inhibitor , partially sensitive to an O15111 ( IKK ) inhibitor , but not sensitive to a Bruton 's tyrosine kinase ( Btk ) inhibitor . Moreover , both the DNA-binding activity and phosphorylation induced by Q9C000 were reduced by the overexpression of a dominant negative Akt in HeLa cells . These results suggest that Q9C000 activates mainly PI3K to phosphorylate p65 and subsequently induces DNA-binding activity of NF-kappaB , independent of its antioxidative function . Inhibition of secretion of interleukin ( IL ) -12/IL-23 family cytokines by 4-trifluoromethyl-celecoxib is coupled to degradation via the endoplasmic reticulum stress protein HERP . Interleukin-12 ( IL-12 ) , p80 , and IL-23 are structurally related cytokines sharing a p40 subunit . We have recently demonstrated that celecoxib and its P35354 -independent analogue 4-trifluoromethyl-celecoxib ( TFM-C ) inhibit secretion but not transcription of IL-12 ( p35/p40 ) and p80 ( p40/p40 ) . This is associated with a mechanism involving altered cytokine-chaperone interaction in the endoplasmic reticulum ( ER ) . In the present study , we found that celecoxib and TFM-C also block secretion of IL-23 ( p40/p19 heterodimers ) . Given the putative ER-centric mode of these compounds , we performed a comprehensive RT-PCR analysis of 23 ER-resident chaperones/foldases and associated co-factors . This revealed that TFM-C induced 1.5-3-fold transcriptional up-regulation of calreticulin , P11021 , P14625 , Q9Y4L1 , P13667 , P30101 , Q9UBS3 , and P30040 . However , more significantly , a 7-fold up-regulation of homocysteine-inducible ER protein ( HERP ) was observed . HERP is part of a high molecular mass protein complex involved in ER-associated protein degradation ( ERAD ) . Using co-immunoprecipitation assays , we show that TFM-C induces protein interaction of p80 and IL-23 with HERP . Both HERP siRNA knockdown and HERP overexpression coupled to cycloheximide chase assays revealed that HERP is necessary for degradation of intracellularly retained p80 by TFM-C . Thus , our data suggest that targeting cytokine folding in the ER by small molecule drugs could be therapeutically exploited to alleviate inappropriate inflammation in autoimmune conditions . DB05773 ( DB05773 ) in human epidermal growth factor receptor 2 ( P04626 ) -positive metastatic breast cancer : latest evidence and clinical potential . In February 2013 , ado-trastuzumab emtansine ( DB05773 , Kadcyla® ) received regulatory approval in the United States for treatment-refractory human epidermal growth factor receptor 2 ( P04626 ) positive metastatic or locally advanced breast cancer based on results from EMILIA , a large phase III trial that compared standard of care lapatinib plus capecitabine to DB05773 . Several other studies have been reported in the metastatic setting and multiple trials are ongoing or planned in the neoadjuvant , adjuvant and advanced disease settings . Here we provide an updated and comprehensive review of clinical trials evaluating DB05773 , discuss management of toxicity associated with this drug , propose potential mechanisms of resistance and offer practical considerations for the treating oncologist . 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 . 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 . DB09045 for the treatment of type 2 diabetes . DB09045 is a novel glucagon-like peptide 1 ( P0C6A0 ) receptor agonist with a unique structure that supports once-weekly dosing in patients with type 2 diabetes ( T2DM ) , most of whom have a big pill burden . It appears to be efficacious in reducing hemoglobin A1c ( HbA1c ) up to 1.59 % and promotes modest weight loss up to 3 kg with a low incidence of hypoglycemia and mild to moderate gastrointestinal adverse events . Convenient weekly dosing could improve compliance and help attain sustained glycemic goals . Addressing obesity is an integral part of T2DM management and weight loss may contribute to better glycemic and cardiovascular benefits . Results of ongoing clinical trials on cardiovascular safety are important to determine the risk-to-benefit ratio . As with any drug , patient selection and ongoing monitoring will be important . If approved , dulaglutide will be one of the first weekly P43220 agonist to be available in a ready-to-use pen device with an automatic injector . A SPECIAL MEETING REVIEW EDITION : Highlights in Anti-Tumor Necrosis Factor Monitoring and Antibody Monitoring From the 2014 DDW Meeting : Digestive Disease Week 2014 May 3-6 , 2014 • Chicago , Illinois : Special Reporting on:• Therapeutic Monitoring of Anti- P01375 Levels and Antibodies to Predict Response and Achieve Mucosal Healing• Prospective Therapeutic Drug Monitoring and Optimization of DB00065 Maintenance Therapy in IBD• Classification of Non- Q9UKU7 , Crohn 's Disease and Ulcerative Colitis in a Young Patient Population Using a Multi-Marker Diagnostic Panel• Persistence of Antibodies to DB00065 for More Than Two Months Predicts Loss of Response to DB00065 in Inflammatory Bowel Diseases• Pre-Operative Serological Markers May Predict Postoperative Crohn 's Disease Recurrence : Results From a Prospective Mono-Centric Trial• Antibodies and Levels of Biologies-Reactive vs Proactive Measurements• Higher DB00352 Nucleotide Concentrations Are Associated With Higher Trough Levels of DB00065 in Patients on Combination Therapy• The Clinical and Immunological Significance of Low Levels of DB00065 in the Absence of Anti-lnfliximab Antibodies in Patients With IBD• Antibodies to DB00051 Predict Inflammation in Crohn 's Patients on Maintenance DB00051 Therapy• Q676U5 Genotype Is Associated With Response to Anti-TNFWith Expert Commentary by:William J. Sandborn , MDProfessor and Chief , Division of Gastroenterology Director , UCSD Q9UKU7 CenterUC San Diego Health SystemLa Jolla , California .

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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 . 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 . 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 . 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 . DB00864 ( FK506 ) increases neuronal expression of P20936 -43 and improves functional recovery after spinal cord injury in rats . DB00864 ( FK506 ) , a widely used immunosuppressant drug , has neurite-promoting activity in cultured PC12 cells and peripheral neurons . The present study investigated whether tacrolimus affects the expression of the neuronal growth-associated protein , P20936 -43 , as well as functional recovery after photothrombotic spinal cord injury in the rat . In injured animals receiving tacrolimus , the number of neurons expressing P20936 -43 mRNA and protein approximately doubled compared to that in injured animals receiving vehicle alone . This increase in P20936 -43-positive cells was paralleled by a significant improvement in neurological function evaluated by open-field and inclined plane tests . Another P62942 ligand ( V-10,367 ) had similar effects on P20936 -43 expression and functional outcome , indicating that the observed effects of tacrolimus do not involve inhibition of the phosphatase calcineurin . Thus , tacrolimus , a drug which is already approved for use in humans , as well as other P62942 ligands which do not inhibit calcineurin , could potentially enhance functional outcome after CNS injury in humans . 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 . Cocaine- and amphetamine-regulated transcript is the neurotransmitter regulating the action of cholecystokinin and leptin on short-term satiety in rats . Vagal CCK-A receptors ( CCKARs ) and leptin receptors ( LRbs ) interact synergistically to mediate short-term satiety . Cocaine- and amphetamine-regulated transcript ( Q16568 ) peptide is expressed by vagal afferent neurons . We sought to demonstrate that this neurotransmitter regulates CCK and leptin actions on short-term satiety . We also examined the signal transduction pathways responsible for mediating the Q16568 release from the nodose ganglia ( NG ) . ELISA studies coupled with gene silencing of NG neurons by RNA interference elucidated intracellular signaling pathways responsible for CCK/leptin-stimulated Q16568 release . Feeding studies followed by gene silencing of Q16568 in NG established the role of Q16568 in mediating short-term satiety . Immunohistochemistry was performed on rat NG neurons to confirm colocalization of CCKARs and LRbs ; 63 % of these neurons contained Q16568 . Coadministration of CCK-8 and leptin caused a 2.2-fold increase in Q16568 release that was inhibited by CCK-OPE , a low-affinity P32238 antagonist . Transfection of cultured NG neurons with steroid receptor coactivator ( P12931 ) or phosphatidylinositol 3-kinase ( PI3K ) small-interfering RNA ( siRNA ) or P40763 lentiviral short hairpin RNA inhibited CCK/leptin-stimulated Q16568 release . Silencing the expression of the P18146 gene inhibited the CCK/leptin-stimulated Q16568 release but had no effect on CCK/leptin-stimulated neuronal firing . Electroporation of NG with Q16568 siRNA inhibited CCK/leptin stimulated c-Fos expression in rat hypothalamus . Feeding studies following electroporation of the NG with Q16568 or P40763 siRNA abolished the effects of CCK/leptin on short-term satiety . We conclude that the synergistic interaction of low-affinity vagal CCKARs and LRbs mediates Q16568 release from the NG , and Q16568 is the principal neurotransmitter mediating short-term satiety . Q16568 release from the NG involves interaction between CCK/ P12931 /PI3K cascades and leptin/ O60674 /PI3K/ P40763 signaling pathways . DB00133 /threonine kinase receptors and ligands . DB00133 /threonine receptors transduce signals for the TGF-beta family , several members of which , such as decapentaplegic and bone morphogenetic proteins , are involved in early patterning of the embryo . The gene encoding the anti-Müllerian hormone ( P03971 ) receptor has recently been cloned ; gene targeting produces the same effects as targeting of the P03971 gene itself . Another divergent member of the TGF-beta family , P39905 , signals through Ret , a tyrosine kinase receptor ; binding to Ret requires the cooperation of GDNFR-alpha . The signal transduction pathway of serine/threonine receptors is now being intensively studied ; the immunophilin P62942 and Q05195 proteins are known to be involved . Cholecystokinin-related peptides , after systemic or central administration , prevent carbon monoxide-induced amnesia in mice . The neuroprotective actions of cholecystokinin ( CCK ) peptides were investigated in a mouse hypoxia model , in which the animals were successively exposed to CO gas . Working memory impairment 5 days after CO exposure was examined by using a Y-maze test ; delayed amnesia was examined 7 days after CO exposure , by using a step-down type passive avoidance test . DB00403 ( 1-100 micrograms/kg , given s.c. 30 min before CO exposure ) significantly prevented the CO-induced impairment of performance in both tests , the improvement being correlated with the severity of hypoxia . This severity was increased by maintaining the body temperature at 38 degrees C . DB00403 was less effective when injected immediately after a single CO exposure . The order of potency of the CCK-peptides administered systemically was : ceruletide > CCK-8S > CCK-8NS >> Q13308 . DB00403 ( 0.03-0.3 micrograms/mouse ) and CCK-8S ( 0.03-1 microgram/mouse ) prevented CO-induced amnesia after i.c.v. administration . Under all experimental conditions , dizocilpine [ MK-801 , (+)-5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate , 500 micrograms/kg s.c. or 10 micrograms/mouse i.c.v. ] prevented completely the CO-induced amnesia . The protective effects of systemic ceruletide were blocked , partially but significantly , by the preadministration of L-364,718 ( 3S-(-)-N- [ 2,3-dihydro-1-methyl-2-oxo-S-phenyl-1H-1,4- benzodiazepine-3-yl ] -1H-indole-2-carboxamide , 1-10 mg/kg i.p. ) , a selective P32238 antagonist . L-365,260 ( [ 3R-(+)-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepine-3-yl ] -N ' - [3-methyl-phenyl]urea ) , a CCK-B antagonist , also decreased ceruletide-induced protection. ( ABSTRACT TRUNCATED AT 250 WORDS ) Signaling by proinflammatory cytokines : oligomerization of TRAF2 and Q9Y4K3 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain . Interleukin-1 ( IL-1 ) and tumor necrosis factor ( P01375 ) stimulate transcription factors AP-1 and NF-kappaB through activation of the Q96HU1 kinases JNK and p38 and the O15111 ( IKK ) , respectively . The P01375 and IL-1 signals are transduced through TRAF2 and Q9Y4K3 , respectively . Overexpressed TRAF2 or Q9Y4K3 activate JNK , p38 , or IKK in the absence of extracellular stimulation . By replacing the carboxy-terminal TRAF domain of TRAF2 and Q9Y4K3 with repeats of the immunophilin P62942 , we demonstrate that their effector domains are composed of their amino-terminal Zn and RING fingers . Oligomerization of the TRAF2 effector domain results in specific binding to Q13233 , a protein kinase capable of JNK , p38 , and IKK activation , and induction of P01375 and IL-1 responsive genes . P01375 also enhances the binding of native TRAF2 to Q13233 and stimulates the kinase activity of the latter . Thus , P01375 and IL-1 signaling is based on oligomerization of TRAF2 and Q9Y4K3 leading to activation of effector kinases . 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 . Short-term weightlessness produced by parabolic flight maneuvers altered gene expression patterns in human endothelial cells . This study focused on the effects of short-term microgravity ( 22 s ) on the gene expression and morphology of endothelial cells ( ECs ) and evaluated gravisensitive signaling elements . ECs were investigated during four German Space Agency ( Deutsches Zentrum für Luft- und Raumfahrt ) parabolic flight campaigns . Hoechst 33342 and acridine orange/ethidium bromide staining showed no signs of cell death in ECs after 31 parabolas ( P31 ) . Gene array analysis revealed 320 significantly regulated genes after the first parabola ( P1 ) and P31 . P29400 , P27658 , P23229 , O75578 , and P05106 mRNAs were down-regulated after P1 . P05305 and Q9NP84 mRNAs were up-regulated . Q9H013 , Q9Y2G2 , P25942 , P06396 , P17252 ( all down-regulated after P1 ) , and Q13131 ( AMPKα1 ) mRNAs ( up-regulated ) provide a very early protective mechanism of cell survival induced by 22 s microgravity . The P42684 gene was significantly up-regulated after P1 and P31 , P07437 was slightly induced , but P62736 and P08670 mRNAs were not changed . β-Tubulin immunofluorescence revealed a cytoplasmic rearrangement . Vibration had no effect . Hypergravity reduced Q9Y2G2 , NOS3 , Q7L8A9 , P50454 ( all P1 ) , P51636 , Q9H013 , Q9NP84 , P25942 , and P23229 ( P31 ) mRNAs . These data suggest that microgravity alters the gene expression patterns and the cytoskeleton of ECs very early . Several gravisensitive signaling elements , such as AMPKα1 and integrins , are involved in the reaction of ECs to altered gravity . O75916 -2 modulates D2 dopamine receptor-mediated Ca2+ channel inhibition in rat striatal cholinergic interneurons . Regulator of G protein signaling ( Q99697 ) proteins negatively regulate receptor-mediated second messenger responses by enhancing the GTPase activity of Galpha subunits . We describe a receptor-specific role for an Q99697 protein at the level of an individual brain neuron . O75916 -2 and Gbeta(5) mRNA and protein complexes were detected in striatal cholinergic and gamma-aminobutyric acidergic neurons . Dialysis of cholinergic neurons with O75916 constructs enhanced basal Ca(2+) channel currents and reduced P14416 modulation of Cav2.2 channels . These constructs did not alter M(2) muscarinic receptor modulation of Cav2.2 currents in the same neuron . The noncatalytic DEP-GGL domain of O75916 antagonized endogenous O75916 -2 activity , enhancing D(2) receptor modulation of Ca(2+) currents . In vitro , O75916 constructs accelerated GTPase activity , in agreement with electrophysiological measurements , and did so more effectively at Go than Gi . These results implicate O75916 -2 as a specific regulator of dopamine receptor-mediated signaling in the striatum and identify a role for P20936 activity modulation by the DEP-GGL domain . The intracellular domain of amyloid precursor protein interacts with P62942 . To elucidate the roles of the P05067 intracellular domain ( AICD ) in the development of Alzheimer 's disease , a yeast two-hybrid system was used to screen for AICD-interacting proteins . Our result revealed that P62942 , an immunophilin with a peptidyl-prolyl cis-trans isomerase ( PPIase ) activity , may interact with AICD . This interaction was confirmed by coimmunoprecipitation studies . P62942 has been shown to be expressed at a higher level in areas of pathology of patients with neurodegenerative diseases . In addition , Pin1 , a member of another PPIase family , has been suggested to be involved in the amyloidogenic P05067 processing and Abeta production . The interaction between P62942 and AICD might hint at a possible role P62942 plays , probably in a fashion similar to Pin1 , in the amyloidogenesis of P05067 . We also found that the interaction was interfered , in a dose-dependent manner , by FK506 , whose neuroprotective effect has been suggested to be correlated with its PPIase inhibitory activity . DB00131 kinase-activated protein kinase ( PRKA ) activators delay meiotic resumption in porcine oocytes . DB00131 -activated kinase ( PRKA ) is a serine/threonine kinase that functions as a metabolic switch in a number of physiological functions . The present study was undertaken to assess the role of this kinase in nuclear maturation of porcine oocytes . RT-PCR and immunoblotting revealed the expression of the Q13131 subunit in granulosa cells , cumulus-oocyte complexes ( COC ) , and denuded oocytes ( DO ) . Porcine COC and DO contained transcripts that corresponded to the expected sizes of the designed primers for Q9Y478 and P54619 . The P54646 subunit was detected in granulosa cells and COC , whereas the Q9UGI9 subunit was not detected in granulosa cells , COC or DO , whereas it was detected in the heart . The Q13131 protein was detected in granulosa cells , COC , DO , and zona pellucida ( ZP ) . In the presence of the pharmacological activator of PRKA 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-monophosphate ( ZMP ) , COC were transiently maintained in meiotic arrest in a fully reversible manner . This inhibitory effect was not observed in DO . Other known PRKA activators , 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside ( AICAR ) and metformin , also blocked meiotic resumption in COC . In contrast to mouse oocytes , in which PRKA activators reverse the inhibitory effect of PDE3 inhibitors , this combination still blocked meiotic resumption in porcine COC . These results demonstrate that the meiotic resumption of porcine COC is transiently blocked by PRKA activators in a dose-dependent manner , and that this effect is dependent on PRKA activity in cumulus cells . The present study describes a new role for PRKA in regulating meiotic resumption in COC and strongly suggests that cumulus cells play an essential role in the control of porcine oocyte maturation through the PRKA metabolic switch . A proliferation switch for genetically modified cells . Receptor dimerization is the key signaling event for many cytokines , including erythropoietin . A system has been recently developed that permits intracellular protein dimerization to be reversibly activated in response to a lipid-soluble dimeric form of the drug FK506 , called FK1012 . FK1012 is used as a pharmacological mediator of dimerization to bring together FK506 binding domains , taken from the endogenous protein P62942 . In experiments reported herein , FK1012-induced dimerization of a fusion protein containing the intracellular portion of the erythropoietin receptor allowed cells normally dependent on interleukin 3 to proliferate in its absence . FK506 competitively reversed the proliferative effect of FK1012 but had no influence on the proliferative effect of interleukin 3 . Signaling pathways activated by FK1012 mimicked those activated by erythropoietin , because both O60674 and P42229 were phosphorylated in response to FK1012 . This approach may provide a means to specifically and reversibly stimulate the proliferation of genetically modified cell populations in vitro or in vivo . 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 . DB00140 ( vitamin B2 ) deficiency impairs P04839 ( Nox2 ) priming and defense against Listeria monocytogenes . DB00140 , also known as vitamin B2 , is converted by riboflavin kinase ( Q969G6 ) into flavin mononucleotide ( Q68DA7 ) and flavin adenine dinucleotide ( DB03147 ) , which are essential cofactors of dehydrogenases , reductases , and oxidases including the phagocytic P04839 ( Nox2 ) . DB00140 deficiency is common in young adults and elderly individuals , who are at the coincidental risk for listeriosis . To address the impact of acute riboflavin deficiency on host defense against Listeria monocytogenes ( L.m. ) , we generated conditional Q969G6 knockout ( KO ) strains of mice . Phagocyte-specific Q969G6 KO impaired the capability of phagocytes to control intracellular L.m. , which corresponded to a greater susceptibility of mice to in vivo challenge with L.m . The oxidative burst of Q969G6 -deficient phagocytes in response to L.m. infection was significantly reduced . Mechanistically , P01375 -induced priming of Nox2 , which is needed for oxidative burst , was defective in Q969G6 -deficient phagocytes . Lack of riboflavin in wild-type macrophages for only 6 h shut down P01375 -induced , Q969G6 -mediated de novo Q68DA7 / DB03147 generation , which was accompanied by diminished ROS production and impaired anti-listerial activity . Vice versa , ROS production by riboflavin-deprived macrophages was rapidly restored by riboflavin supplementation . Our results suggest that acute riboflavin deficiency immediately impairs priming of Nox2 , which is of crucial relevance for an effective phagocytic immune response in vivo . Protein-protein interactions : an application of Tus-Ter mediated protein microarray system . In this chapter , we present a novel , cost-effective microarray strategy that utilizes expression-ready plasmid DNAs to generate protein arrays on-demand and its use to validate protein-protein interactions . These expression plasmids were constructed in such a way so as to serve a dual purpose of synthesizing the protein of interest as well as capturing the synthesized protein . The microarray system is based on the high affinity binding of Escherichia coli " Tus " protein to " Ter , " a 20 bp DNA sequence involved in the regulation of DNA replication . The protein expression is carried out in a cell-free protein synthesis system , with rabbit reticulocyte lysates , and the target proteins are detected either by labeled incorporated tag specific or by gene-specific antibodies . This microarray system has been successfully used for the detection of protein-protein interaction because both the target protein and the query protein can be transcribed and translated simultaneously in the microarray slides . The utility of this system for detecting protein-protein interaction is demonstrated by a few well-known examples : Jun/Fos , FRB/ P62942 , p53/ Q00987 , and P11802 /p16 . In all these cases , the presence of protein complexes resulted in the localization of fluorophores at the specific sites of the immobilized target plasmids . Interestingly , during our interactions studies we also detected a previously unknown interaction between P24941 and p16 . Thus , this Tus-Ter based system of protein microarray can be used for the validation of known protein interactions as well as for identifying new protein-protein interactions . In addition , it can be used to examine and identify targets of nucleic acid-protein , ligand-receptor , enzyme-substrate , and drug-protein interactions . Human epidermal Langerhans ' cells are targets for the immunosuppressive macrolide tacrolimus ( FK506 ) . BACKGROUND : The immunosuppressive macrolide tacrolimus ( FK506 ) has been shown to inhibit allergic contact dermatitis in animal models as well as in human beings . More recently , successful treatment of atopic dermatitis with an ointment containing tacrolimus has been reported . OBJECTIVES : We explored the effects of this compound on epidermal Langerhans ' cells ( LCs ) , which are known to play an important pathophysiologic role in inflammatory skin diseases . METHODS : The expression of the intracellular FK506 binding protein ( P62942 ) was monitored on freshly isolated and cultured epidermal LCs . Phenotyping and functional exploration of LCs treated with different concentrations of tacrolimus and beta-methasone valerate ( betaMv ) were performed . RESULTS : P62942 is expressed in freshly isolated LCs but is lost while they are maturating into mature dendritic cells . DB00864 inhibited the expression of IL-2R ( CD25 ) and of the costimulatory molecules P33681 ( P33681 .1 ) and P25942 . Expression of MHC class I and II was also affected , whereas P42081 ( P33681 .2 ) expression was not altered . In contrast , betaMv strongly increased the expression of CD25 . Paradoxically , while decreasing P25942 and MHC class I expression , betaMv significantly increased the expression of MHC class II , P33681 , and P42081 on cultured LCs but impaired their allostimulatory activity . DB00864 was about 100 times more potent than betaMv at inhibiting LC stimulatory function . CONCLUSION : DB00864 can exert immunopharmacologic alterations on LCs , which may account , at least in part , for the therapeutic effect of this compound in eczematous skin diseases .

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Angiotensin II and basic fibroblast growth factor mitogenic pathways in human fetal mesangial cells . Angiotensin II ( Ang II ) and basic fibroblast growth factor ( P09038 / P09038 ) play relevant roles in renal development . Since the signaling pathways modulating the mitogenic effects of Ang II and P09038 in human fetal mesangial cells ( HFMc ) are not clearly defined , we carried out experiments to determine whether they would exert their mitogenic effects by modulating the activity of the mitogen-activated protein kinases ( MAPK ) [ extracellular signal-regulated kinase-2 ( P28482 ) ] and DB02527 signaling pathways . In confluent HFMc , P09038 ( 20 ng/mL ) induced a significant 4-fold increase in P28482 activity and [ 3H ] -thymidine incorporation ( 6-fold ) . In contrast , under similar tissue culture conditions , Ang II ( 10(-6) M ) induced a more modest increase in P28482 activity ( 2-fold ) and [ 3H ] -thymidine incorporation ( 35 +/- 4 % ) . The mitogen-activated protein kinase kinase-1 ( MEK-1 ) inhibitor PD098059 ( 25 microM ) almost completely abolished the P09038 -induced proliferation in HFMc but did not significantly affect Ang II proliferative effects . In the presence of the DB02527 elevating agent isoproterenol , Ang II and P09038 induced opposite changes in DB02527 accumulation and cell growth . DB01064 inhibited the basal and P09038 -induced proliferation of HFMc through a MEK-1/2-independent pathway that included the accumulation of DB02527 . In contrast , isoproterenol increased Ang II mitogenic effects in correlation with a reduction in DB02527 accumulation . We conclude that Ang II and P09038 modulate the proliferation of HFMc through the stimulation of different MEK-1/2-dependent and independent signaling pathways . Activation of MEK-1/2 is required but not sufficient for mitogenesis in HFMc . The accumulation of DB02527 in HFMc counteracts the mitogenic effects of P09038 by a MEK-1/2-independent pathway . The genetics of complex cholestatic disorders . Cholestatic liver diseases are caused by a range of hepatobiliary insults and involve complex interactions among environmental and genetic factors . Little is known about the pathogenic mechanisms of specific cholestatic diseases , which has limited our ability to manage patients with these disorders . However , recent genome-wide studies have provided insight into the pathogenesis of gallstones , primary biliary cirrhosis , and primary sclerosing cholangitis . A lithogenic variant in the gene that encodes the hepatobiliary transporter Q9H221 has been identified as a risk factor for gallstone disease ; this variant has been associated with altered cholesterol excretion and metabolism . Other variants of genes encoding transporters that affect the composition of bile have been associated with cholestasis , namely O95342 , which encodes the bile salt export pump , and P21439 , which encodes hepatocanalicular phosphatidylcholine floppase . In contrast , studies have associated primary biliary cirrhosis and primary sclerosing cholangitis with genes encoding major histocompatibility complex proteins and identified loci associated with microbial sensing and immune regulatory pathways outside this region , such as genes encoding IL12 , Q14765 , Q13568 , P60568 and its receptor ( IL2R ) , P10747 , and P33681 . These discoveries have raised interest in the development of reagents that target these gene products . We review recent findings from genetic studies of patients with cholestatic liver disease . Future characterization of genetic variants in animal models , stratification of risk alleles by clinical course , and identification of interacting environmental factors will increase our understanding of these complex cholestatic diseases . Ectonucleotidases P49961 and CD73 on OvCA cells are potent adenosine-generating enzymes responsible for adenosine receptor 2A-dependent suppression of T cell function and NK cell cytotoxicity . The ectonucleotidases P49961 and CD73 degrade immune stimulatory DB00171 to adenosine that inhibits T and NK cell responses via the A(2A) adenosine receptor ( P29274 ) . This mechanism is used by regulatory T cells ( T(reg) ) that are associated with increased mortality in OvCA . Immunohistochemical staining of human OvCA tissue specimens revealed further aberrant expression of P49961 in 29/36 OvCA samples , whereas only 1/9 benign ovaries showed weak stromal P49961 expression . CD73 could be detected on 31/34 OvCA samples . While 8/9 benign ovaries also showed CD73 immunoreactivity , expression levels were lower than in tumour specimens . Infiltration by P01730 (+) and CD8(+) T cells was enhanced in tumour specimens and significantly correlated with P49961 and CD73 levels on stromal , but not on tumour cells . In vitro , human OvCA cell lines SK-OV-3 and OaW42 as well as 11/15 ascites-derived primary OvCA cell cultures expressed both functional P49961 and CD73 leading to more efficient depletion of extracellular DB00171 and enhanced generation of adenosine as compared to activated T(reg) . Functional assays using siRNAs against P49961 and CD73 or pharmacological inhibitors of P49961 , CD73 and P29274 revealed that tumour-derived adenosine inhibits the proliferation of allogeneic human P01730 (+) T cells in co-culture with OvCA cells as well as cytotoxic T cell priming and NK cell cytotoxicity against SK-OV3 or OAW42 cells . Thus , both the ectonucleotidases P49961 and CD73 and P29274 appear as possible targets for novel treatments in OvCA , which may not only affect the function of T(reg) but also relieve intrinsic immunosuppressive properties of tumour and stromal cells . A pilot study to evaluate the effects of P05155 on the toxicity of high-dose interleukin 2 . In a pilot study six patients received 4 days ' treatment with interleukin 2 ( P60568 ) [ cumulative dose ( CD ) 264 +/- 26 x 10(6) IU m-2 ] and P05155 ( DB05341 ) ( loading dose 2,000 U , followed by 500-1,000 U twice daily ) . Toxicity was compared with that in patients given 4 days ' treatment with standard ( CD 66 +/- 12 x 10(6) IU m-2 ) or escalating-dose ( CD 99 +/- 8 x 10(6) IU m-2 ) P60568 . P60568 -induced hypotension was equivalent and complement activation was less after P60568 + DB05341 ( C3a = 10.5 +/- 3.2 nmol l-1 ) than following standard ( 14.1 +/- 8.4 nmol l-1 ) or escalating-dose ( 18.3 +/- 2.9 nmol l-1 ) P60568 . This study demonstrates that DB05341 administration during P60568 treatment is safe and warrants further study to evaluate its ability to ameliorate P60568 -induced toxicity . 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 . Gastrointestinal prokinetic drugs have different affinity for the human cardiac human ether-à-gogo K(+) channel . Agonists of the serotonin 5-hydroxytryptamine 4 ( Q13639 ) receptor are widely used to activate motility in the gastrointestinal tract . Among these , cisapride was recently withdrawn from the U.S. market because of its proarrhythmic effects . Cisapride is a potent blocker of human ether-à-gogo ( Q12809 ) K(+) channels and prolongs the cardiac action potential in a reverse use dependence manner . We compared the effects of four different Q13639 receptor agonists ( cisapride , prucalopride , renzapride and mosapride ) on cloned Q12809 channels with the objective to evaluate and compare their proarrhythmic potential . K(+) currents from Q12809 -transfected COS-7 cells were recorded under physiological conditions using the whole cell configuration of the patch-clamp technique . Short ( 500 ms ) depolarizing prepulses were used and following deactivating Q12809 currents were measured . Cisapride inhibited the Q12809 channels in a concentration-dependent manner with an IC(50) of 2.4 10(-7) M . The IC(50) value for prucalopride to block Q12809 ( 5.7 10(-6) M ) was 20-fold higher than that of cisapride . DB04917 was slightly more potent than prucalopride ( IC(50) = 1.8 10(-6) M ) . Mosapride produced no significant effects on the recombinant Q12809 current . The voltage dependence of Q12809 block was also investigated . The block mediated by cisapride or renzapride was voltage-dependent whereas that produced by prucalopride was not . We conclude that the rank order of potency of Q13639 agonists to block Q12809 is cisapride > renzapride > prucalopride > mosapride . We also conclude that Q13639 agonists devoid of side effects on the Q12809 current such as mosapride can be found as a safe alternative to cisapride . The selective 5-hydroxytryptamine (5-HT)4-receptor agonist RS67506 enhances lower intestinal propulsion in mice . Interactions of gastrointestinal prokinetic benzamides with 5-hydroxytryptamine (5-HT)3 and Q13639 receptors and the relation to their effects on gastrointestinal propulsion were investigated . DB04917 and zacopride potently inhibited 5- Q9H205 -receptor-mediated contractions in the guinea pig colon , whereas RS67506 ( 1-(4-amino-5-chloro-2-methoxyphenyl)-3- [ 1- ( 2-methyl sulphonylamino ) ethyl-4-piperidinyl ] -1-propanone hydrochloride ) , a selective Q13639 -receptor agonist , showed no inhibition . RS67506 , renzapride and zacopride all exerted Q13639 receptor-mediated relaxation in the carbachol-precontracted rat oesophagus . In mice , RS67506 shortened the whole gut transit time , whereas renzapride and zacopride were reported to prolong it . Gastrointestinal prokinetic benzamides , which are selective for Q13639 -receptor agonistic over 5- Q9H205 -receptor antagonistic action , may be useful in treating gastrointestinal disorders associated with impaired lower intestinal propulsion such as constipation . Prevention of acute and chronic allograft rejection by a novel retinoic acid receptor-alpha-selective agonist . To investigate the involvement of retinoic acid receptor ( RAR ) -alpha in allograft rejection , we investigated the effect of a novel selective agonist to the receptor , ER-38925 , in a mouse cardiac allograft model . Prophylactic treatment with ER-38925 inhibited the acute rejection of the mouse cardiac allograft ( BALB/c --> C3H/HeN ) at 0.3 and 3 mg/kg , and its effect was enhanced in combination with tacrolimus . In this model , ER-38925 remarkably inhibited cytotoxic T lymphocyte induction and alloantigen-stimulated production of cytokines , i.e. P60568 , IL-12 and P01579 . In the chronic rejection model , combined treatment with tacrolimus and ER-38925 reduced the grade and incidence of arteriosclerosis in the cardiac allografts significantly more potently than tacrolimus monotherapy . ER-38925 inhibited the proliferation of rat aortic smooth muscle cells stimulated in vitro , presumably through the induction of a cyclin-dependent kinase inhibitor , p27(kip-1) . Those results provide a rationale for using P10276 agonists as immunosuppressants in human organ transplantation . Program death-1 engagement upon TCR activation has distinct effects on costimulation and cytokine-driven proliferation : attenuation of Q9Y6W8 , P05112 , and Q9HBE4 , but not P10747 , P13232 , and P40933 responses . The program death 1 ( P18621 ) receptor and its ligands , P18621 ligand (PD-L)1 and Q9BQ51 , define a novel regulatory pathway with potential inhibitory effects on T , B , and monocyte responses . In the present study , we show that human P01730 (+) T cells express P18621 , Q9NZQ7 , and Q9BQ51 upon activation , and Abs to the receptor can be agonists or antagonists of the pathway . Under optimal conditions of stimulation , Q9Y6W8 but not P10747 costimulation can be prevented by P18621 engagement . P60568 levels induced by costimulation are critical in determining the outcome of the P18621 engagement . Thus , low to marginal P60568 levels produced upon Q9Y6W8 costimulation account for the greater sensitivity of this pathway to P18621 -mediated inhibition . Interestingly , exogenous P60568 , P13232 , and P40933 but not P05112 and Q9HBE4 can rescue P18621 inhibition , suggesting that among these cytokines only those that activate P42229 can rescue P18621 inhibition . As P42229 has been implicated in the maintenance of IL-2Ralpha expression , these results suggest that P13232 and P40933 restore proliferation under conditions of P18621 engagement by enhancing high-affinity IL-2R expression and hence , P60568 responsiveness . Definition of functional domains in P135gag-myb-ets and p48v-myb proteins required to maintain the response of neuroretina cells to basic fibroblast growth factor . The v-myb- and v-ets-containing E26 retrovirus induces the proliferation of chicken neuroretina ( P21554 ) cells in minimal medium . Proliferation of E26 P21554 cells is strongly stimulated by basic fibroblast growth factor ( P09038 ) . The v-myb-containing avian myeloblastosis virus also induces the proliferation of infected P21554 cells stimulated by P09038 . Both E26 P21554 and avian myeloblastosis virus P21554 cells are able to form colonies in soft agar in the presence of P09038 . This suggests that the v-myb product , a nuclear sequence-specific DNA-binding protein which activates gene expression in transient transfection assays , plays a role in the proliferative response of the infected P21554 cells . To determine the structure-function relationships of P135gag-myb-ets and p48v-myb , we have used deletion mutants expressed in retroviral vectors and have analyzed their effect on P21554 cell proliferation as well as their effect on the P21554 cell response to P09038 . We show that v-ets is not required for P09038 stimulation but increases the proliferation of P21554 cells in minimal medium . In the v-myb mutants , the gag sequences derived from the helper virus increase the potency of the myb gene . The carboxyl-terminal domain required for the growth and transformation of myeloid cells and needed for maximal trans-activation in transient DNA transfection assays in fibroblasts was not required for the growth and P09038 response of P21554 cells . In contrast , the domain encompassing amino acids 240 to 301 ( containing part of the transcriptional activation domain of v-myb ) was absolutely required for the response of P21554 cells to P09038 and could be functionally replaced by the carboxyl-terminal transcriptional activation domain of the VP16 protein of herpes simplex virus . DB01064 inhibits fibroblast growth factor-2-induced growth of renal epithelial cells . The signal transduction pathways modulating P09038 effects in renal tubular epithelial cells ( RTEc ) are not completely understood . Since the DB02527 and the mitogen-activated protein kinase ( MAPK ) pathways can modulate the growth of RTEc , we studied whether two DB02527 elevating agents , isoproterenol and 8-bromo- DB02527 , would modulate basic fibroblast growth factor ( P09038 ) induction of MAPK activity ( P28482 ) and cell proliferation in human renal proximal tubular epithelial cells ( RPTEc ) and Madin-Darby canine kidney cells ( MDCK clone EI1 ) . DB01064 , but not P09038 , stimulated DB02527 production in RPTEc and MDCKEI1 cells . P09038 , isoproterenol , and 8-bromo- DB02527 alone increased P28482 activity in both cell types . However , isoproterenol and 8-bromo- DB02527 partially inhibited the P09038 induction of P28482 activity , but only isoproterenol inhibited the proliferation of both cell types . PD098059 ( 25 microM ) , an inhibitor of MAPK kinase ( Q02750 /2 ) , blocked the P09038 mitogenic effects , but did not affect the 8-bromo- DB02527 -induced mitogenic effects in MDCKEI1 cells . These findings suggest that activation of P28482 is required but not sufficient for mitogenesis in RTEc . We conclude that isoproterenol inhibits the growth-promoting effects of P09038 in RTEc via MEK-dependent and -independent pathways . 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 . CP-101,606 , a potent neuroprotectant selective for forebrain neurons . The neuroprotective activity of ( 1S,2S ) -1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol ( CP-101,606 ) , an N-methyl-D-aspartate ( DB01221 ) receptor antagonist structurally similar to ( (+/-)-(R*,S*)-alpha-(4-hydroxyphenyl)-beta-methyl-4-(phenylmethyl)-1-++ +piperidineethanol ( ifenprodil ) , was investigated in neurons in primary culture . CP-101,606 potently and efficaciously protected hippocampal neurons from glutamate toxicity but was > 900-fold less effective for cerebellar granule neurons . The neuroprotective activity in the hippocampal neurons is mediated through a high affinity binding site distinct from the agonist and thienylcyclohexylpiperidine ( DB01520 ) binding sites of the DB01221 receptor . Autoradiography indicates the CP-101,606 binding site is localized in forebrain , most notably in hippocampus and the outer layers of cortex . The functional selectivity for hippocampal neurons , forebrain localization of binding sites , and structural relation to ifenprodil suggest that CP-101,606 is an DB01221 antagonist highly selective for Q13224 subunit containing receptors . Differential role of the basolateral amygdala 5- Q9H205 and Q13639 serotonin receptors upon ACPA-induced anxiolytic-like behaviors and emotional memory deficit in mice . BACKGROUND AND AIM : The critical role of cannabinoidergic and serotonergic systems of the amygdala in modulation of anxiety-like behaviors and emotional memory has already been demonstrated . The present study aimed to investigate the possible role of the basolateral amygdala ( BLA ) 5- Q9H205 and Q13639 serotonergic systems upon ACPA ( P21554 cannabinoid receptor agonist ) -induced anxiolytic-like behaviors and emotional memory impairment using the elevated plus-maze ( EPM ) test-retest paradigm in male mice . METHOD : bilateral guide-cannulae were implanted to allow intra-BLA microinjection of serotonergic agents . RESULTS : the intraperitoneal injection of ACPA could induce anxiolytic-like behaviors and reduce the emotional memory formation . Intra-BLA injection of M-Chlorophenylbiguanide ( M-Chl , a 5- Q9H205 serotonin receptor agonist ) neither altered the anxiety-like behaviors nor the emotional memory formation by itself , while the higher dose of Y-25130 ( a 5- Q9H205 serotonin receptor antagonist ) reduced the emotional memory formation and locomotor activity but not the anxiety-like behaviors . Furthermore , injection of a higher dose of RS67333 and RS23597 ( as Q13639 serotonin receptor agonist and antagonist , respectively ) did not alter the anxiety-like behaviors , while reduced the emotional memory formation . In addition , the intra-BLA injection of M-Chl but not Y-25130 and RS67333 restored the ACPA-induced anxiolytic-like behaviors and emotional memory deficit , while a higher dose of RS67333 decreased the locomotor activity . Moreover , the intra-BLA microinjection of RS23597 could restore the ACPA-induced anxiolytic-like behaviors but not the emotional memory deficit . CONCLUSION : based on our findings , ACPA seems to induce its anxiolytic-like behaviors and emotional memory formation deficits via activation and deactivation of the BLA Q13639 and 5- Q9H205 serotonin receptors . 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 . Molecular mechanisms governing different pharmacokinetics of ginsenosides and potential for ginsenoside-perpetrated herb-drug interactions on Q9NPD5 . BACKGROUND AND PURPOSE : Ginsenosides are bioactive saponins derived from Panax notoginseng roots ( Sanqi ) and ginseng . Here , the molecular mechanisms governing differential pharmacokinetics of 20(S)-protopanaxatriol-type ginsenoside Rg1 , ginsenoside Re and notoginsenoside Q96GN5 and 20(S)-protopanaxadiol-type ginsenosides Rb1 , Rc and Rd were elucidated . EXPERIMENTAL APPROACH : Interactions of ginsenosides with human and rat hepatobiliary transporters were characterized at the cellular and vesicular levels . A rifampin-based inhibition study in rats evaluated the in vivo role of organic anion-transporting polypeptide (Oatp)1b2 . Plasma protein binding was assessed by equilibrium dialysis . Drug-drug interaction indices were calculated to estimate potential for clinically relevant ginsenoside-mediated interactions due to inhibition of human OATP1Bs . KEY RESULTS : All the ginsenosides were bound to human Q9NPD5 and rat Oatp1b2 but only the 20(S)-protopanaxatriol-type ginsenosides were transported . Human multidrug resistance-associated protein (MRP)2/breast cancer resistance protein ( Q9UNQ0 ) /bile salt export pump ( O95342 ) /multidrug resistance protein-1 and rat Mrp2/Bcrp/Bsep also mediated the transport of the 20(S)-protopanaxatriol-type ginsenosides . Glomerular-filtration-based renal excretion of the 20(S)-protopanaxatriol-type ginsenosides was greater than that of the 20(S)-protopanaxadiol-type counterparts due to differences in plasma protein binding . DB01045 -impaired hepatobiliary excretion of the 20(S)-protopanaxatriol-type ginsenosides was effectively compensated by the renal excretion in rats . The 20(S)-protopanaxadiol-type ginsenosides were potent inhibitors of Q9NPD5 . CONCLUSION AND IMPLICATIONS : Differences in hepatobiliary and in renal excretory clearances caused markedly different systemic exposure and different elimination kinetics between the two types of ginsenosides . Caution should be exercised with the long-circulating 20(S)-protopanaxadiol-type ginsenosides as they could induce hepatobiliary herb-drug interactions , particularly when patients receive long-term therapies with high-dose i.v. Sanqi or ginseng extracts . Regulation of mouse inducible costimulator ( Q9Y6W8 ) expression by Fyn- Q13469 and P29323 signaling in T cells . The inducible costimulator ( Q9Y6W8 ) , a member of the P10747 family of costimulatory molecules , is rapidly induced upon T cell activation . Although the critical role of Q9Y6W8 in costimulating T cell responses is well documented , little is known of the intracellular signaling pathways and mechanisms that regulate Q9Y6W8 expression . Here , we report that Fyn , NFAT , and P29323 signaling influence Q9Y6W8 expression as various chemical inhibitors , such as Q99463 that targets Src kinases , U0126 that targets Q02750 /2 , and cyclosporin A or FK506 that targets calcineurin and thereby affects NFAT , attenuate T cell receptor-mediated Q9Y6W8 induction . Moreover , ectopic expression of Q13469 or a constitutively active P36507 amplifies Q9Y6W8 transcription and transactivates a 288-bp core region of the icos promoter in luciferase reporter assays . We also identify a site on the icos promoter that is sensitive to P29323 signaling and further show that Q13469 can bind the icos promoter in vivo and that this binding is diminished when Fyn signaling is ablated . The normal activation of P29323 but reduced nuclear translocation of Q13469 in Fyn(-/-) P01730 (+) T cells further suggest that Fyn and Q13469 act in a common axis , separate from that involving P29323 , to drive Q9Y6W8 transcription . Taken together , our findings indicate that Fyn-calcineurin- Q13469 and P36507 - P27361 /2 are two independent signaling pathways that cooperate to control T cell receptor-mediated Q9Y6W8 induction . New-generation Q13639 receptor agonists : potential for treatment of gastrointestinal motility disorders . IMPORTANCE OF THE FIELD : Gastrointestinal ( GI ) dysmotility is an important mechanism in functional GI disorders ( FGIDs ) including constipation , irritable bowel syndrome , functional dyspepsia , and gastroparesis . 5-hydroxytryptamine(4) ( 5-HT(4) ) receptors are targets for the treatment of GI motility disorders . However , older 5-HT(4) receptor agonists had limited clinical success because they were associated with changes in the function of the cardiac Q12809 potassium channel . AREAS COVERED IN THIS REVIEW : We conducted a PubMed search using the following key words alone or in combination : 5-HT(4) , safety , toxicity , pharmacokinetics , pharmacodynamics , clinical trial , cardiac , hERG , arrhythmia , potassium current , elderly , prucalopride , ATI-7505 , and velusetrag ( DB05655 ) , to review mechanisms of action , clinical efficacy , safety and tolerability of three new-generation 5-HT(4) receptor agonists . WHAT THE READER WILL GAIN : DB06480 , ATI-7505 , and velusetrag ( DB05655 ) are highly selective , high-affinity 5-HT(4) receptor agonists that are devoid of action on other receptors within their therapeutic range . Their efficacy has been demonstrated in pharmacodynamic studies which demonstrate acceleration of colonic transit and , to a variable degree , in clinical trials that significantly relieve chronic constipation . Currently available evidence shows that the new 5-HT(4) receptor agonists have safe cardiac profiles . TAKE HOME MESSAGE : New-generation 5-HT(4) receptor agonists and future drugs targeting organ-specific splice variants are promising approaches to treat GI dysmotility , particularly colonic diseases . The effect of genetic polymorphisms of P10276 gene on the adverse effects profile of isotretinoin-treated acne patients . OBJECTIVE : DB00982 is a vitamin A-derived medication that is associated with significant adverse effects including arthralgia/ myalgia , nose bleeds , headache , dyslipidemia , liver dysfunction and depression . The mechanism for development of such adverse effects remains elusive , and it is not known why adverse effects develop only in some patients . In this study , we examined the association between rs9303285 , rs2715554 and rs4890109 genetic polymorphisms in the retinoic acid receptor alpha ( P10276 ) , one of the main targets of isotretinoin , and the adverse effects of oral isotretinoin therapy . MATERIALS AND METHODS : Clinical adverse effects data were collected from patient file and by patient interview . Lipids and liver enzymes were measured in blood samples collected from acne patients ( n = 300 ) at baseline and during oral isotretinoin treatment . P10276 polymorphisms were genotyped using polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP ) . RESULTS : Three-locus haplotype ( Rs2715554 C/T - Rs4890109 G/T - Rs9303285 T/C ) analysis showed that frequencies of CTG and TTG haplotypes are significantly associated with occurrence of arthralgia , myalgia , nose bleeds and headache in patients treated with isotretinoin . In addition , TCG haplotype was associated with nose bleeds and headache , whereas TTT haplotype was associated with arthralgia and myalgia . Furthermore , levels of Q9NRA2 were increased , and Q9NRA2 % change was more , after 1 month of treatment in patients with the TC genotype of rs2715554 polymorphism . Finally , allele T of rs9303285 was found to be protective against developing depression in the patients treated with isotretinoin . CONCLUSIONS : Our findings suggest an association between polymorphisms of P10276 gene and some of some common adverse effects of oral isotretinoin . Multiple structural determinants of voltage-dependent magnesium block in recombinant DB01221 receptors . The voltage-dependent block of DB01221 channels by Mg2+ is an important functional element of DB01221 receptors , since relief of block by depolarization plays a key role in some forms of ischemic neurodegeneration and synaptic plasticity . To identify the relevant structural domains responsible for block by Mg2+ and DB01520 , we used site-directed mutagenesis to change individual amino acids of the rat NR1A subunit in a transmembrane region ( 599-DALTLSSAMWFSWGVLLNSGIGE-621 , mutated residues underlined ) previously shown to donate residues that influence ionic selectivity . Ten mutant NR1A subunits were co-expressed in Xenopus oocytes with either the epsilon 1 or Q12879 subunits , and receptor properties were analyzed under two-electrode voltage clamp . The mutation N616R virtually abolished voltage-dependent Mg2+ block , reduced DB01593 block 5-fold and greatly reduced Ba2+ permeability in confirmation of previous reports . This mutation also reduced the potency of DB01520 as a use-dependent blocker by 200-fold . The remaining low-affinity DB01520 block did not appear to be use-dependent , suggesting two blocking sites for DB01520 . None of the other mutations differed significantly from NR1A itself except S617N , which displayed a 6-fold reduction in Mg2+ block . A well-barrier model of permeation through the DB01221 receptor channel is presented that quantitatively reproduces voltage-dependent Mg2+ block . This model demonstrates that only minimum changes energy profiles experienced by permeating ions , equivalent to the energy of a single hydrogen or ionic bond , are required to abolish Mg2+ block . These findings indicate that only small structural changes are needed to convert a Mg(2+)-insensitive ion channel to a channel with pronounced voltage-dependent Mg2+ block . Role of nitric oxide in the expression of hepatic vascular stress genes in response to sepsis . This study examined the role of nitric oxide ( NO ) on the expression of the hepatic vasoregulatory gene during polymicrobial sepsis . DB02533 ( AG , 100 mg/kg ) or Nomega-nitro-L-arginine methyl ester ( L-NAME , 100 mg/kg ) was injected intraperitoneally at 0 , 3 , 6 , 10 , and 20 h after a cecal ligation and puncture ( CLP ) . The heart rate increased 24 h after the CLP , and this increase was attenuated by L-NAME and further attenuated by AG . The mean arterial pressure in the CLP animals did not change significantly 24 h after the onset of sepsis but was increased after the L-NAME injection . Sepsis increased the serum aminotransferase levels , which were attenuated by AG but augmented by L-NAME . CLP increased the mRNA level of the ET-1 and ETB receptors in the liver . This increase was prevented by AG but augmented by L-NAME . The level of P35228 and P09601 mRNA expression were increased by CLP , which was prevented by both AG and L-NAME . The level of P01375 and P35354 mRNA expression increased after CLP , and was attenuated by AG . These results show that P35228 and P29474 are regulated differently in sepsis . While P29474 appears to have a protective role in liver microcirculation , the strong upregulation of P35228 might contribute to a microvascular dysfunction and hepatic injury . DB00640 A(2A) receptor gene ( P29274 ) variants may increase autistic symptoms and anxiety in autism spectrum disorder . Autism spectrum disorders ( ASDs ) are heterogeneous disorders presenting with increased rates of anxiety . The adenosine A(2A) receptor gene ( P29274 ) is associated with panic disorder and is located on chromosome 22q11.23 . Its gene product , the adenosine A(2A) receptor , is strongly expressed in the caudate nucleus , which also is involved in P51689 . As autistic symptoms are increased in individuals with 22q11.2 deletion syndrome , and large 22q11.2 deletions and duplications have been observed in P51689 individuals , in this study , 98 individuals with P51689 and 234 control individuals were genotyped for eight single-nucleotide polymorphisms in P29274 . Nominal association with the disorder was observed for rs2236624-CC , and phenotypic variability in P51689 symptoms was influenced by rs3761422 , rs5751876 and rs35320474 . In addition , association of P29274 variants with anxiety was replicated for individuals with P51689 . Findings point toward a possible mediating role of P29274 variants on phenotypic expression in P51689 that need to be replicated in a larger sample . 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 . Modular competition driven by DB01221 receptor subtypes in spike-timing-dependent plasticity . N-methyl-d-aspartate receptors ( NMDARs ) play a critical role in transducing neuronal activity patterns into changes in synaptic strength . However , how they mediate this transduction in response to physiological stimuli has remained elusive . In particular , it has been debated whether different NMDAR subtypes play opposing signaling roles in synaptic plasticity . Using perforated patch-clamp recordings from pairs of synaptically connected glutamatergic neurons in dissociated hippocampal culture , we found that spike-timing-dependent potentiation induced by pairing pre- and postsynaptic spikes required the activation of a fast component of NMDAR current that is likely to be mediated by Q12879 -containing NMDARs ( Q12879 -NRs ) . In contrast , spike-timing-dependent depression required a slow component of NMDAR current carried by Q13224 -containing NMDARs ( Q13224 -NRs ) . CV analysis showed that the locus of this depression was primarily presynaptic in pairs of cells making strong synaptic connections , whereas weaker synapses showed no clear preference for pre- or postsynaptic expression . This depression was not significantly reduced by antagonism of the P21554 receptor , in contrast to spike-timing-dependent depression in the neocortex that requires presynaptic P21554 signaling . With blockade of Q13224 -NRs , spike triplets that contained both potentiating and depressing spike-timing components induced net potentiation . However , when the putative Q12879 -NR population is inhibited , these spike triplets resulted in either depression or no net change , depending on the temporal order of the spike-timing components . These results imply a dynamic competition between signaling modules that can be biased by differentially antagonizing NMDAR subtypes during the induction of spike-timing-dependent plasticity . Using a simple model , we show that such a modular competition recapitulates our observations . [ Effects of salvia miltiorrhiza injection on gentamicin-induced expression of nitric oxide synthase isoforms in guinea pig cochlea ] . OBJECTIVE : To investigate the effects of salvia miltiorrhiza injection ( SM ) on gentamicin ( GM ) -induced expression of nitric oxide synthase ( NOS ) isoforms in guinea pig cochlea , and to explore the protective mechanism of SM on GM-induced ototoxicity . METHODS : 40 guinea pigs were randomly assigned to 4 groups : control group , GM group , SM group and GM plus SM group . Expression of NOS isoforms in the guinea pig cochlea was detected by the SABC method of immunohistochemistry and microscope image analysis technique . Auditory threshold was tested by auditory brainstem response ( Q12979 ) measurement . RESULTS : P35228 ( P35228 /NOS II ) expression and Q12979 threshold in GM plus SM group were both significantly declined as compared with those in GM group ( P < 0.01 ) . Moreover , change of P35228 expression was in high correlation with that of Q12979 threshold ( [ r ] > 0.7 , P < 0.01 ) . While expression of neuronal NOS ( P29475 /NOS I ) and endothelial NOS ( P29474 / NOS III ) showed no significant differences in all groups . CONCLUSION : SM had no effect on the expression of P29475 and P29474 , but could inhibit P35228 high-expression induced by GM to reduce excessive generation of NO , therefore SM could protect against GM ototoxicity . Long-term observations on the reversibility of cochlear dysfunction after transient ischemia . To examine the reversibility of functional damage to the cochlea after transient ischemia , cochlear ischemia of 0-60 min was induced in 34 albino guinea pigs . Thresholds of auditory brainstem response ( Q12979 ) were then followed for 5 days after ischemia . Although the Q12979 threshold returned to almost the pre-ischemic value after 15 min ischemia , ischemia of 30 and 60 min duration induced irreversible dysfunction . DB02533 , an inducible NO synthase ( P35228 ) inhibitor , significantly ameliorated the post-ischemic cochlear dysfunction induced by 60 min ischemia . Morphological findings of the hair cells were consistent with these functional results . These results indicate that ischemia of 30 min or longer induces irreversible damage to the cochlea and that P35228 plays injury-producing roles in this type of injury . Increased interleukin 21 and follicular helper T-like cells and reduced interleukin 10+ B cells in patients with new-onset systemic lupus erythematosus . OBJECTIVE : To elucidate the potential role of follicular helper T cells ( TFH ) and interleukin 10 ( P22301 )+ B cells in the development of systemic lupus erythematosus ( SLE ) . METHODS : The numbers of peripheral blood P26842 + , P28907 + , P42081 + , CD95+ , P22301 + B cells , and inducible T cell costimulator ( Q9Y6W8 )+ , programmed death-1 ( P18621 )+ , Q9HBE4 + , P32302 + P01730 + TFH-like cells were examined in 23 patients with new onset SLE and 20 healthy controls ( HC ) . RESULTS : In comparison with HC , significantly reduced numbers of P15391 + and P22301 + B cells , but increased numbers of P26842 (high) , P42081 + , CD95+ B cells , P32302 + P01730 + , Q9Y6W8 + , P18621 + , and Q9HBE4 + TFH-like cells were detected , which were accompanied by higher levels of serum Q9HBE4 , but lower levels of P22301 in the patients . Treatment with anti-SLE therapy modulated the imbalance of different subsets of B and TFH-like cells . The levels of serum Q9HBE4 and P22301 were positively correlated with the numbers of P01730 + P32302 + TFH-like and P15391 + P06127 +CD1d+ B cells in the patients , respectively . The numbers of P26842 (high) B cells were correlated positively with Q9HBE4 + TFH-like cells , but negatively with P22301 + B cells . The values of SLE Disease Activity Index , P01024 , and erythrocyte sedimentation rate were correlated positively with serum Q9HBE4 , but negatively with P22301 in those patients . CONCLUSION : Our data indicate that the imbalance of Q9HBE4 + TFH-like , P26842 (high) , and P22301 + B cells may be associated with the pathogenesis of SLE , and levels of serum Q9HBE4 and P22301 may be valuable for evaluating disease activity in SLE . Endocannabinoid-dependent neocortical layer-5 LTD in the absence of postsynaptic spiking . Long-term depression ( LTD ) was induced in neocortical layer 5 pyramidal connections by pairing presynaptic firing with subthreshold postsynaptic depolarization ( dLTD ) or via a spike-timing protocol ( tLTD ) . Like tLTD , dLTD reduced short-term depression and increased the coefficient of variation consistent with a presynaptic site of expression . Also like tLTD , dLTD was blocked by P21554 cannibinoid receptor blockade and required activation of presumably presynaptic Q13224 -containing N-methyl-D-aspartate receptors . The two forms of LTD had identical magnitudes and time courses and occluded one another , and neither depended on frequency . Finally , dLTD shares with tLTD the asymmetric temporal window of induction . In conclusion , the types of LTD induced by these two protocols are indistinguishable , suggesting that the mechanism that underlies tLTD paradoxically does not require postsynaptic spiking : The subthreshold postsynaptic depolarizations of dLTD appear to fully substitute for postsynaptic spiking . 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 . The Retinoic Acid Receptor-alpha mediates human T-cell activation and Th2 cytokine and chemokine production . BACKGROUND : We have recently demonstrated that all-trans-retinoic acid ( DB00755 ) and 9-cis-retinoic acid ( 9-cis RA ) promote P05112 , P05113 and P35225 synthesis , while decreasing P01579 and P01375 expression by activated human T cells and reduces the synthesis of IL-12p70 from accessory cells . Here , we have demonstrated that the observed effects using DB00755 and 9-cis RA are shared with the clinically useful RAR ligand , 13-cis retinoic acid ( 13-cis RA ) , and the retinoic acid receptor-alpha ( P10276 ) -selective agonist , AM580 but not with the P10826 /gamma ligand , 4-hydroxyphenylretinamide ( DB05076 ) . RESULTS : The increase in type 2 cytokine production by these retinoids correlated with the expression of the T cell activation markers , Q07108 and P28907 . The P10276 -selective agonist , AM580 recapitulated all of the T cell activation and type 2 cytokine-inducing effects of DB00755 and 9- DB00982 , while the P10276 -selective antagonist , RO 41-5253 , inhibited these effects . CONCLUSION : These results strongly support a role for P10276 engagement in the regulation of genes and proteins involved with human T cell activation and type 2 cytokine production . Recombinant human P05155 for the treatment of hereditary angioedema due to C1 inhibitor deficiency ( DB05341 -HAE ) . The lack of C1 inhibitor function that results in excessive production of bradykinin causing the angioedema seen in hereditary angioedema ( HAE ) is well established . Several drugs have been developed to treat and prevent attacks in patients suffering from HAE due to C1 inhibitor deficiency ( DB05341 -HAE ) . Plasma-derived C1INH has been used to replace the deficiency of C1 inhibitor ( C1INH ) and has been approved for both treatment of attacks and for prophylactic therapy to prevent attacks . P03952 inhibitor ( ecallantide ) and bradykinin receptor antagonist ( icatibant ) are both effective for treatment of acute attacks , but their short half-life limits the use for prophylaxis . Androgens , in particular danazol , are effective for long-term prophylaxis , but adverse event profile can limit its use . Recombinant C1 inhibitor derived from transgenic rabbits has recently been approved for use in treatment of DB05341 -HAE attacks and is effective and appears safe with minimal adverse event profile . 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 . Hereditary angioedema : a Taiwanese family with a novel gene mutation . Hereditary angioedema ( HAE ) is an autosomal dominant disorder caused by a deficiency of P05155 ( DB05341 ) . Affected individuals have attacks of swelling involving almost any part of the body . We studied a family with 15 living members , including a 16-year-old girl who had 3 attacks of angioedema in 2 years . Her paternal uncle had died of asphyxiation during an attack 15 years previously . We analyzed the blood of each family member for P01024 , C4 , and DB05341 levels and sequenced the P05155 ( formerly P05155 ) gene that codes for DB05341 . Six individuals had decreased serum levels of C4 and DB05341 , and they were all found to have a single nucleotide A deletion at codon 210 of the gene , 1210fsX210 , a novel mutation that accounts for the HAE in this family . 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 . 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 .

MH_train_1511

MH_train_1511

MH_train_1511

interacts_with DB00215?

multiple_choice

Nox2 and Nox4 mediate tumour necrosis factor-α-induced ventricular remodelling in mice . Reactive oxygen species ( ROS ) and pro-inflammatory cytokines are crucial in ventricular remodelling , such as inflammation-associated myocarditis . We previously reported that tumour necrosis factor-α ( P01375 -α ) -induced ROS in human aortic smooth muscle cells is mediated by NADPH oxidase subunit Nox4 . In this study , we investigated whether P01375 -α-induced ventricular remodelling was mediated by Nox2 and/or Nox4 . An intravenous injection of murine P01375 -α was administered to a group of mice and saline injection was administered to controls . Echocardiography was performed on days 1 , 7 and 28 post-injection . Ventricular tissue was used to determine gene and protein expression of Nox2 , Nox4 , P01160 , interleukin ( IL ) -1β , P60568 , P05231 , P01375 -α and to measure ROS . Nox2 and Nox4 siRNA were used to determine whether or not Nox2 and Nox4 mediated P01375 -α-induced ROS and upregulation of IL-1β and P05231 in adult human cardiomyocytes . Echocardiography showed a significant increase in left ventricular end-diastolic and left ventricular end-systolic diameters , and a significant decrease in the ejection fraction and fractional shortening in mice 7 and 28 days after P01375 -α injection . These two groups of mice showed a significant increase in ventricular ROS , P01160 , IL-1β , P60568 , P05231 and P01375 -α proteins . Nox2 and Nox4 mRNA and protein levels were also sequentially increased . ROS was significantly decreased by inhibitors of NADPH oxidase , but not by inhibitors of other ROS production systems . Nox2 and Nox4 siRNA significantly attenuated P01375 -α-induced ROS and upregulation of IL-1β and P05231 in cardiomyocytes . Our study highlights a novel P01375 -α-induced chronic ventricular remodelling mechanism mediated by sequential regulation of Nox2 and Nox4 subunits . In vivo P61073 expression , lymphoid cell phenotype , and feline immunodeficiency virus infection . Primary isolates of feline immunodeficiency virus ( FIV ) appear to require binding to CD134 in conjunction with P61073 (X4) to infect P60568 -dependent T-cell-derived cells in culture . However , much less is known about the role of X4 for the infection of cells in vivo . To investigate the correlation between X4 expression and FIV infection in cats acutely infected with FIV-C-Pgmr we used high-speed fluorescence-activated cell sorting and realtime PCR to co-analyze cell phenotypes from lymph node , thymus , bone marrow and blood for FIV infection and X4 expression . X4 expression was greatest in lymph node , both in frequency and in mean fluorescence intensity . The thymus demonstrated a higher proviral burden in X4+ thymic T cells ( approximately 14 % in X4+ thymic T cells and 7 % in X4- cells ) whereas , proviral loads were similar between X4+ and X4- cell populations in all other tissues examined . Assuming a minimum of one proviral copy per cell , a maximum of approximately 50 % of FIV-positive cells were X4+ . The highest fraction of FIV-infected X4- cells was present in bone marrow . Regardless of X4 status , proviral loads were higher in lymph node and blood T cells than in B cells . These studies provide both a positive association between X4 expression and FIV infection and introduce the probability that X4-independent infection occurs in other target cells in vivo . 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 . Cytokine regulation of intercellular adhesion molecule-1 ( P05362 ) expression on human glioblastoma cells . Intercellular adhesion molecule-1 ( P05362 ) has recently been identified as one of the ligands for lymphocyte function-associated antigen-1 ( LFA-1 ) . Immunohistochemical staining of frozen tissue sections using the P05362 antibody P23921 /1 demonstrated significant levels of P05362 expression on human glioblastoma cells and on intratumoural vascular endothelial cells . P05362 was weakly expressed or absent from low grade gliomas and absent from normal and fetal brain . P05362 expression was similar to that of MHC class II . HLA-DR antigens . Glioblastoma cell lines constitutively expressed P05362 to a minimal or moderate extent . Surface antigen expression of P05362 and P05362 -specific mRNA could be significantly increased by incubating glioblastoma cells with interleukin-1 beta ( P01584 ) , tumour necrosis factor-alpha ( P01375 ) , and interferon-gamma ( P01579 ) . P60568 , P05112 , P05231 and transforming growth factor beta 2 ( TGF-beta 2 ) had no significant effect on surface antigen expression . Significant enhancement of P05362 expression was obtained using P01375 and P01584 at 1-10 U/ml and at 500 U/ml of P01579 . Induction of P05362 specific mRNA was observed 4 h after cytokine treatment and decreased by 24 h . Surface antigen expression of P05362 increased for up to 48 h after treatment . Effects of a novel P08908 receptor agonist , E4424 , on gastric adherent mucus levels following restraint stress in rats . Several novel arylpiperazine serotonin 1A receptor agonists , developed as anxiolytics , have antisecretory and gastroprotective effects in rats . E4424 ( 2-¿4-[4-(4-chloropyrazol-1-yl)butyl]-1-piperazinyl ¿pyrimidine ; DB04970 dihydrochloride ) , has potent anti-gastric secretory and antiulcer effects . Preliminary data indicated an enhancing effect of E4424 on gastric mucus that may underlie its gastroprotective actions . We therefore tested the effects of acute and chronic administration of E4424 and of a reference P08908 receptor agonist , 8-OHDPAT [ 8-hydroxy-2-(di-n-propylamino)tetralin ] , on gastric mucus levels in rats subjected to cold-restraint stress , a procedure associated with depletion of gastric mucus and the development of mucosal injury . Acute oral administration of E4424 increased adherent mucus levels by 12 % , 11 % , and 13 % , relative to controls . Chronic E4424 significantly increased gastric mucus relative to controls ( 69 % increase ) . Acute oral treatment with 8-OHDPAT did not affect gastric mucus level . Acute intraperitoneal 8-OHDPAT slightly increased mucus levels . Chronic twice per day 8-OHDPAT did not affect mucus levels ; however , chronic once per day treatment with 8-OHDPAT significantly elevated gastric mucus levels at the highest doses used . For E4424 , there is a strong correlation between reduction of gastric mucosal injury and increase in gastric mucus level , suggesting that the action of E4424 on glandular mucus levels is an important mechanism underlying its gastroprotective effects . Identification of three adjacent amino acids of interleukin-2 receptor beta chain which control the affinity and the specificity of the interaction with interleukin-2 . The beta chain of the interleukin-2 ( P60568 ) receptor ( IL-2R beta ) and the interleukin-3 ( P08700 ) binding protein AIC2A are members of the family of cytokine receptors , which also includes the receptors for growth hormone ( P10912 ) and prolactin . A four amino acid sequence of AIC2A has recently been shown to be critical for P08700 binding . We analyze here the function of the analogous sequence of human IL-2R beta and identify three amino acids , Ser132 , His133 and Tyr134 , which play a critical role in P60568 binding . We show that some mutant P60568 proteins with substitutions of a critical DB00128 residue in the N-terminal alpha-helix bind the mutant IL-2R beta receptor with a higher affinity than the wild-type receptor . This suggests that the critical Asp34 in the ligand and the sequence DB00133 - DB00117 - DB00135 ( positions 132-134 ) in the receptor interact directly . On the double barrel beta-stranded structural model of cytokine receptors , the residues important for ligand binding in IL-2R beta , AIC2A and P10912 map to strikingly similar locations within a barrel , with the interesting difference that it is the N-terminal barrel for P10912 and the C-terminal barrel for IL-2R beta and AIC2A . Differential gene expression of the three natriuretic peptides and natriuretic peptide receptor subtypes in human liver . BACKGROUND : Various effects of atrial natriuretic peptide ( P01160 ) on the liver have been observed . However , there is limited information about the types of receptors for natriuretic peptides expressed by the human liver . AIM : To investigate gene expression of the three NP receptor types ( NPR ) as well as of the NP in human liver . METHODS : Presence of mRNA coding for all three NPR and for P01160 , brain and P23582 ( DB04899 , P09543 ) was investigated by reverse transcription-polymerase chain reaction ( RT-PCR ) . Human liver tissues and hepatocellular carcinoma tissues were examined . RESULTS : Specific PCR products for all three NPR , namely P16066 , B , and C , could be detected . Moreover , P01160 and P09543 , but not DB04899 mRNA was detectable . The concentration of P01160 transcripts was up to fivefold higher in hepatocellular carcinoma compared with non-tumorous liver tissue of the same subjects . No difference in the expression of NP receptors relative to P04406 mRNA of tumorous and non-tumorous tissue was observed except of slightly increased P16066 transcripts . CONCLUSION : These data show that NPR transcripts are coexpressed with P01160 and P09543 mRNA in the human liver . This provides evidence for a local NP system in the human liver . 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 . 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 . Novel use of endogenous GH-measurement directly after transsphenoidal microsurgery in acromegaly treated with pegvisomant . OBJECTIVE : The P10912 antagonist pegvisomant is increasingly used as therapy in acromegaly . Pituitary surgery might be indicated on pegvisomant treatment , due to side effects , adenoma growth or intention to cure after primary treatment . This study was initiated to clarify if , and when , GH measurement could be useful postoperatively with an assay specific for endogenous GH that does not cross-react with pegvisomant . METHODS : This study was designed as a prospective study in 2006 with the German Pituitary Working Group . Only 2 cases with potentially resectable adenomas from the German DB00082 Observational Study ( GPOS ) had been operated . Now with a post-operative follow-up of more than 5 years in these 2 cases , the usefulness of immediate pre-operative GH measurement shortly after pegvisomant treatment was evaluated . RESULTS : In both patients a steep decline of endogenous GH after transnasal microsurgery could be proven by using the special GH assay after near radical or radical removal , of the GH secreting adenomas respectively . Conventional GH assays showed no effect . GH half-life was more than 20 min in the patient with a small invasive residual adenoma and less than 20 min in the cured patient . Endogenous GH-levels declined to less than 1 ng/ml in the days after surgery in the patient with long-term cure . CONCLUSION : Measurement of endogenous GH in this special subgroup of patients under pegvisomant therapy can be used to decide upon early reoperation . Thus the beneficial effect of pegvisomant on acromegalic symptoms can be kept without interfering with post-operative monitoring of GH levels . 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 . Inorganic arsenic impairs proliferation and cytokine expression in human primary T lymphocytes . Inorganic arsenic is a toxic environmental contaminant to which humans are mainly exposed through drinking water . This metalloid impairs functions of several key immune cells . Particularly , it reduces P60568 secretion and proliferation of blood peripheral mononuclear cells stimulated by lectins that , however , do not mimic physiological T cell activation . The present study used isolated human T cells activated , in a more physiological manner , through stimulation with CD3/ P10747 antibodies , to carefully analyze the impact of arsenic on T cell proliferation and cytokine expression . We demonstrate that non cytotoxic concentrations of sodium arsenite ( As(III) , 0.25-2μM ) significantly reduce T cell proliferation by increasing the percentage of non dividing cells blocked in P55008 phase and by preventing cyclin D3 and P30304 expression . They also markedly , although not totally , reduces P60568 expression at both mRNA and protein levels ; however , metalloid-dependent inhibition of T cells could not be reversed by addition of recombinant P60568 . In addition , As(III) markedly reduces secretion of interferon-γ without impairing that of P05112 and P35225 ; it also decreases interferon-γ mRNA levels but increases those of P35225 . Finally , simultaneously to its immune effects , As(III) rapidly and potently increases expression of the redox-sensitive genes P09601 , P15559 and P48507 in activated T cells without altering the levels of reactive oxygen species . In conclusion , our results demonstrate that As(III) inhibits T cell proliferation , independently of P60568 , and alters the Th balance of cytokines secreted by co-stimulated T cells which thus constitute direct targets of this major environmental contaminant . P60568 inhibits DB01221 receptor-mediated currents directly and may differentially affect subtypes . Using whole-cell patch-clamp recordings , this study investigated the effects of interleukin-2 ( P60568 ) on N-methyl-d-aspartate ( DB01221 ) receptor-mediated currents ( I( DB01221 ) ) in rat cultured hippocampal neurons and human embryonic kidney ( P29320 ) 293 cells expressing recombinant DB01221 receptors . We found that P60568 ( 0.01-1ng/ml ) immediately and significantly decreased peak I( DB01221 ) in cultured neurons . Interestingly , the peak I( DB01221 ) induced in P29320 293 cells was also inhibited by P60568 . We also found that P60568 differentially decreased the peak amplitudes of Q12879 - and Q13224 -containing DB01221 receptor-mediated currents ( I( Q12879 ) and I( Q13224 ) ) by 54+/-5 % and 30+/-4 % , respectively . These results provide new evidence that P60568 induces rapid inhibition of peak currents of DB01221 receptor-mediated responses with possible Q9UHB4 / Q12879 and Q9UHB4 / Q13224 subtype-differentiation , and suggest that the inhibition is mediated by direct interaction between P60568 and DB01221 receptors . Expression of the human concentrative nucleotide transporter 1 ( O00337 ) gene correlates with clinical response in patients affected by Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) undergoing a combination treatment with 2-chloro-2'-deoxyadenosine ( DB00242 ) and DB00073 . PURPOSE : Resistance to nucleoside analogues agents is likely to be multifactorial and could involve a number of mechanisms affecting drug penetration , metabolism and targeting . In vitro studies of resistant human cell lines have confirmed that human concentrative nucleoside transporter 1 ( O00337 ) -deficient cells display resistance . EXPERIMENTAL DESIGN : We applied real-time PCR method to assess the mRNA expression of equilibrative and concentrative nucleoside transporter ( hENT1 , O00337 ) , deoxycytidine and deoxyguanosine kinase ( P27707 , Q16854 ) , 5'-nucleotidase ( 5'-NT ) , ribonucleotide reductase catalytic and regulatory ( P23921 , P31350 ) subunits in bone marrow cells from 32 patients with Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) who received 2CdA-based chemotherapy . Responses to chemotherapy , were then correlated to the expression of these markers . RESULTS : All 32 patients enrolled expressed lower levels of O00337 as compared to healthy donors . In univariate analysis , lower expression level of O00337 ( p=0.0021 ) and P31350 ( p=0.02 ) correlated with response to chemotherapy . In particular , patients with low levels of O00337 achieved inferior clinical response . No significant correlation between these genes expression and age , stage of disease was found . This study suggests that nucleotidase expression levels can be used to identify subgroups of WM and SLL patients who will likely respond differently to a 2CdA-based therapy . Molecular analysis of Ras activation by tyrosine phosphorylated Vav . Vav has been shown to activate Ras ( 1-3 ) and is regulated by tyrosine phosphorylation ( 1 ) or binding of diglycerides ( 3 ) to the cysteine rich domain . In the present study employing different Ras activation assay techniques using [3H]GDP release or [32P]alpha GTP-binding from membrane-bound or soluble recombinant Ras , we demonstrate that Ras activity can be increased by tyrosine phosphorylated Vav upon cellular stimulation via the P60568 receptor or the TCR/CD3-complex . Increase of [32P]alpha GTP-binding to Ras catalyzed by phosphorylated Vav is similar to the activity of immunoprecipitated Sos . The activity of Vav measured by binding of [32P]alpha GTP to Ras was linear with respect to the concentration of Vav protein used . To study molecular characteristics of this Vav-Ras interaction , we used several Ras mutants and demonstrate that Vav activity towards Ras depends on the integrity of the same or similar domains as Ras activation by P18827 25 or CDC 25 . 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 . Initial characterization of the glutamate-cysteine ligase modifier subunit Gclm(-/-) knockout mouse . Novel model system for a severely compromised oxidative stress response . Glutamate-cysteine ligase ( GCL ) is the rate-limiting enzyme in the DB00143 biosynthesis pathway . In higher eukaryotes , this enzyme is a heterodimer comprising a catalytic subunit ( P48506 ) and a modifier subunit ( P48507 ) , which change the catalytic characteristics of the holoenzyme . To define the cellular function of P48507 , we disrupted the mouse Gclm gene to create a null allele . Gclm(-/-) mice are viable and fertile and have no overt phenotype . In liver , lung , pancreas , erythrocytes , and plasma , however , DB00143 levels in Gclm(-/-) mice were 9-16 % of that in Gclm(+/+) littermates . DB00151 levels in Gclm(-/-) mice were 9 , 35 , and 40 % of that in Gclm(+/+) mice in kidney , pancreas , and plasma , respectively , but remained unchanged in the liver and erythrocytes . Comparing the hepatic GCL holoenzyme with P48506 in the genetic absence of P48507 , we found the latter had an approximately 2-fold increase in K(m) for glutamate and a dramatically enhanced sensitivity to DB00143 inhibition . The major decrease in DB00143 , combined with diminished GCL activity , rendered Gclm(-/-) fetal fibroblasts strikingly more sensitive to chemical oxidants such as H(2)O(2) . We conclude that the Gclm(-/-) mouse represents a model of chronic DB00143 depletion that will be very useful in evaluating the role of the P48507 subunit and DB00143 in numerous pathophysiological conditions as well as in environmental toxicity associated with oxidant insult . 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 . Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 -induced survival . This inhibition can not be overcome by increasing concentrations of P05113 and is not due to the blocking of Na+ channels by lidocaine . Here we report that one class of K+ channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 . In contrast , increasing concentrations of P08700 or granulocyte-macrophage P04141 overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 -sensitive K+ channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [3H]glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases . Letter of retraction : Dual effect of DB06809 , a P61073 antagonist , on bleomycin-indiced lung inflammation .

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Neutralization of botulinum neurotoxin by a human monoclonal antibody specific for the catalytic light chain . BACKGROUND : Botulinum neurotoxins ( BoNT ) are a family of category A select bioterror agents and the most potent biological toxins known . Cloned antibody therapeutics hold considerable promise as BoNT therapeutics , but the therapeutic utility of antibodies that bind the BoNT light chain domain ( LC ) , a metalloprotease that functions in the cytosol of cholinergic neurons , has not been thoroughly explored . METHODS AND FINDINGS : We used an optimized hybridoma method to clone a fully human antibody specific for the LC of serotype A BoNT ( DB00083 ) . The 4LCA antibody demonstrated potent in vivo neutralization when administered alone and collaborated with an antibody specific for the HC . In Neuro-2a neuroblastoma cells , the 4LCA antibody prevented the cleavage of the DB00083 proteolytic target , P60880 . Unlike an antibody specific for the HC , the 4LCA antibody did not block entry of DB00083 into cultured cells . Instead , it was taken up into synaptic vesicles along with DB00083 . The 4LCA antibody also directly inhibited DB00083 catalytic activity in vitro . CONCLUSIONS : An antibody specific for the DB00083 LC can potently inhibit DB00083 in vivo and in vitro , using mechanisms not previously associated with BoNT-neutralizing antibodies . Antibodies specific for BoNT LC may be valuable components of an antibody antidote for BoNT exposure . 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 . Immunohistochemical analysis of low-grade and high-grade prostate carcinoma : relative changes of parathyroid hormone-related protein and its parathyroid hormone 1 receptor , osteoprotegerin and receptor activator of nuclear factor-kB ligand . AIM : To investigate multiple bone cytokines produced by prostate carcinoma ( PCa ) as a novel strategy to differentiate potential aggressiveness in localised PCa using immunohistochemical analysis . METHODS : A total of 47 cases of PCa undergoing radical prostatectomy or transurethral prostatic resection at our institution ( Fundación Jiménez Díaz ( Grupo Capio ) , Madrid , Spain ) between January 1991 and June 1998 were identified as low-grade ( < or =4 ; n = 22 ) or high-grade ( > or =7 , excluding 7 ( 3+4 ) cases ; n = 25 ) PCa according to Gleason grade . PCa specimens were immunostained for : parathyroid hormone ( PTH ) -related protein ( P12272 ) , the Q03431 , osteoprotegerin and receptor activator of nuclear factor-kappa B ligand ( O14788 ) , as well as Ki67 ( a proliferation marker ) and P28906 ( an angiogenesis marker ) . RESULTS : PCa samples showed an increased immunostaining for both osteoprotegerin and O14788 , associated with tumour grade and P12272 positivity , in the tumoral epithelium . Using a score value of 4-corresponding to moderate staining - as cut-off , the best sensitivity value was for P12272 ( with C-terminal antiserum P13671 ; 100 % ) ; wheras the best specificity value was for O14788 ( 95 % ) . CONCLUSIONS : All the evaluated factors are overexpressed mainly in the high-grade tumours . Our findings indicate that , in most patients with PCa ( with Ki67 values between 1 % and 9 % ) , sequential determination of C-terminal P12272 and O14788 immunoreactivities is a useful approach to discriminate low-grade and high-grade tumours . 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 . Comparison of patient intake of ticagrelor , prasugrel , or clopidogrel on restoring platelet function by donor platelets . BACKGROUND : Bleeding complications are a common side effect in patients under dual antiplatelet ( anti- Q02083 ) therapy . Q02083 transfusion provides a treatment option for these patients . However it is currently unclear if , and to what extent , Q9H244 inhibitors influence Q02083 function of donor PLTs and if patients taking these medications are likely to benefit from Q02083 transfusions . STUDY DESIGN AND METHODS : We investigated the effect of blood and plasma of clopidogrel- , prasugrel- , and ticagrelor-treated patients on Q02083 function of blood from healthy volunteers in flow cytometry , light transmission aggregometry , and multiple electrode aggregometry ( MEA ) . RESULTS : Our results demonstrate that clopidogrel had no and prasugrel had only mild effects on donor Q02083 function , but the reversible Q9H244 inhibitor ticagrelor completely abolished adenosine diphosphate-mediated Q02083 activation in all assays tested . We further show that ticagrelor itself and not elevated adenosine concentrations in patient plasma were responsible for the observed effects . Moreover , we show that a modified MEA assay could provide a simple and rapid tool to allow determination of whether patients are likely to benefit from Q02083 transfusions . CONCLUSION : Our results provide novel insights into potential differences between the Q9H244 inhibitors on donor Q02083 function in an in vitro setting , which may provide implications for future Q02083 transfusion strategies in these patients . DB00887 blocks P13569 GCl in the native sweat duct . DB00887 is well known for its ability to inhibit the nonconductive Na+-K+-2Cl- cotransporter . We were surprised in preliminary studies to find that bumetanide in the contraluminal bath also inhibited NaCl absorption in the human sweat duct , which is apparently poor in cotransporter activity . Inhibition was accompanied by a marked decrease in the transepithelial electrical conductance . Because the cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channel is richly expressed in the sweat duct , we asked whether bumetanide acts by blocking this anion channel . We found that bumetanide 1 ) significantly increased whole cell input impedance , 2 ) hyperpolarized transepithelial and basolateral membrane potentials , 3 ) depolarized apical membrane potential , 4 ) increased the ratio of apical-to-basolateral membrane resistance , and 5 ) decreased transepithelial Cl- conductance ( GCl ) . These results indicate that bumetanide inhibits P13569 GCl in both cell membranes of this epithelium . We excluded bumetanide interference with the protein kinase A phosphorylation activation process by " irreversibly " phosphorylating P13569 [ by using adenosine 5'-O-(3-thiotriphosphate) in the presence of a phosphatase inhibition cocktail ] before bumetanide application . We then activated P13569 GCl by adding 5 mM DB00171 . DB00887 in the cytoplasmic bath ( 10(-3) M ) inhibited approximately 71 % of this DB00171 -activated P13569 GCl , indicating possible direct inhibition of P13569 GCl . We conclude that bumetanide inhibits P13569 GCl in apical and basolateral membranes independent of phosphorylation . The results also suggest that > 10(-5) M bumetanide can not be used to specifically block the Na+-K+-2Cl- cotransporter . 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 . Genetically rescued tetrahydrobiopterin-depleted mice survive with hyperphenylalaninemia and region-specific monoaminergic abnormalities . One of the possibly mutated genes in DOPA-responsive dystonia ( DRD , Segawa 's disease ) is the gene encoding P30793 , which is the rate-limiting enzyme for tetrahydrobiopterin ( BH4 ) biosynthesis . Based on our findings on 6-pyruvoyltetrahydropterin synthase ( Q03393 ) gene-disrupted ( Pts(-/-) ) mice , we suggested that the amount of tyrosine hydroxylase ( TH ) protein in dopaminergic nerve terminals is regulated by the intracellular concentration of BH4 . In this present work , we rescued Pts(-/-) mice by transgenic introduction of human Q03393 cDNA under the control of the dopamine beta-hydroxylase promoter to examine regional differences in the sensitivity of dopaminergic neurons to BH4-insufficiency . The DPS-rescued ( Pts(-/-) , DPS ) mice showed severe hyperphenylalaninemia . Human Q03393 was efficiently expressed in noradrenergic regions but only in a small number of dopaminergic neurons . DB03886 and dopamine contents , and TH activity in the striatum were poorly restored compared with those in the midbrain . TH-immunoreactivity in the lateral region of the striatum was far weaker than that in the medial region or in the nucleus accumbens . We concluded that dopaminergic nerve terminals projecting to the lateral region of the striatum are the most sensitive to BH4-insufficiency . Biochemical and pathological changes in DPS-rescued mice were similar to those in human malignant hyperphenylalaninemia and DRD . Plasticity of neurohypophysial terminals with increased hormonal release during dehydration : ultrastructural and biochemical analyses . DB00067 - ( AVP ) and oxytocin- ( P01178 ) secreting magnocellular neurons undergo gross structural changes with chronic physiological stimulation . Here , we investigated subcellular aspects of plasticity in rat neurohypophysial terminals during dehydration . Ultrastructural analyses demonstrated that chronic dehydration by 2 % NaCl drinking for 7 days significantly decreased the numbers of neurosecretory granules and microvesicles but not the numbers of mitochondria . Moreover , in dehydrated rats , terminals making neurovascular contacts enlarged , whereas terminals in apposition to astrocytes , i.e. , neuroglial contacts , became smaller . Western blot analyses demonstrated significant decreases in the levels of P13726 and Thy-1 together with those of AVP- and P01178 -neurophysin , but the levels of synaptophysin , P60880 , and P20936 -43 were unchanged . Both P13726 and Thy-1 were recovered in the buffer-insoluble pellet , and phosphatidyl inositol-specific phospholipase C treatment released both molecules from the crude membrane fraction , indicating that they are attached to terminal membranes by glycosylphosphatidyl inositol anchors . Confocal microscopic observations demonstrated that P13726 colocalized with Thy-1 in the same terminals of magnocellular neurons . In contrast , the level of calretinin , a Ca(2+) binding protein was significantly increased with chronic dehydration . Thus , the present results suggest that enhancement of neurovascular contacts results from rearrangement of terminal-astrocyte and terminal-vessel contacts rather than enlargement or sprouting of magnocellular terminals themselves . The down-regulation of P13726 and Thy-1 may contribute to enhancement of neurovascular contacts that accompany increased peptide release during dehydration . [ 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 . P09917 pathway promotes cell proliferation in human glioma cell lines . OBJECTIVE : P09917 ( P09917 ) is a key enzyme in the synthesis of leukotrienes ( LTs ) , that might promote carcinogenesis . We investigated P09917 expression and examined whether the P09917 pathway is associated with the proliferation of human brain tumors . METHODS : We immunohistochemically evaluated the profile of P09917 expression in various types of brain tumors obtained from 42 patients , and examined the proliferative effects of the P09917 pathway in human glioma cell lines using a proliferation assay . RESULTS : Immunohistochemistry of glioblastomas , astrocytomas , meningiomas , medulloblastomas , craniopharyngiomas , ependymomas , neurinomas , oligodendrogliomas , malignant lymphomas , dysembryoplastic neuroepithelial and metastatic brain tumors revealed P09917 expression in the cytoplasm and nuclei or nuclear envelopes of tumor cells . The P09917 inhibitor A861 and the P09960 hydrolase inhibitor DB03424 dose-dependently suppressed the proliferation of A172 cells , a glioma cell line . CONCLUSIONS : We confirmed the expression of P09917 in various human brain tumors and demonstrated the partial suppression of tumor growth by inhibitors of the P09917 - P09960 hydrolase pathway in human glioma cell lines . The P09917 - P09960 pathway might play roles in the proliferation of human glioma cells . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . Brain transplantation of human neural stem cells transduced with tyrosine hydroxylase and P30793 provides functional improvement in animal models of Parkinson disease . Parkinson disease is a neurodegenerative disease characterized by loss of midbrain dopaminergic neurons resulting in movement disorder . Neural stem cells ( NSC ) of the CNS have recently aroused a great deal of interest , not only because of their importance in basic research of neural development , but also for their therapeutic potential in neurological disorders . We have recently generated an immortalized human NSC cell line , HB1. P13726 , via retrovirus-mediated v-myc transfer . This line is capable of self-renewal , is multipotent , and expresses cell specific markers for NSC , DB00171 -binding cassettes transporter ( Q9UNQ0 ) and nestin . Next , we co-transduced the P13726 NSC line with genes encoding tyrosine hydroxylase ( TH ) and P30793 ( GTPCH1 ) in order to generate dopamine-producing NSC . The P13726 .TH.GTPCH human NSC line expresses TH and GTPCH phenotypes as determined by RT-PCR , western blotting and immunocytochemistry , and shows a 800 to 2000-fold increase in production of L-dihydroxyphenyl alanine in HPLC analysis . A marked improvement in amphetamine-induced turning behavior was observed in parkinsonian rats implanted with P13726 .TH.GTPCH cells , but not in control rats receiving P13726 NSC . In the animals showing functional improvement , a large number of TH-positive P13726 .TH.GTPCH NSC were found at injection sites . These results indicate that human NSC , genetically transduced with TH and GTPCH1 genes , have great potential in clinical utility for cell replacement therapy in patients suffering from Parkinson disease . [ Eicosanoids as mediators in ARDS ] . Eicosanoids partly develop from the metabolism of arachidonic acid through the cyclooxygenase or the lipoxygenase pathway . Lipoxygenase products are the leukotrienes ( P09960 , LTB4 , LTC4 , LTD4 , LTE4 ) and the 5-hydroxyeicosatetraenoic acid ( 5-HETE ) . Cyclooxygenase products are the prostanoids ( prostaglandins [ PG ] D2 , E2 , F2 , I2 and thromboxane A2 ) . The other part of the eicosanoids develops from the metabolism of two other fatty acids over the same pathways ; 8,11,14-eicosatrienoic acid leads to the prostaglandins D1 , E1 , F1 , I1 and the leukotrienes A3 , B3 , P01024 , D3 , E3 . From 5,8,11,14,17-eicosapentaenoic acid result the prostaglandins D3 , E3 , P13726 , I3 and the leukotrienes A5 , P46977 , P01031 , D5 , O94989 . The pathophysiological changes in ARDS are mainly due to an imbalance of opposing effects of mediators . In this regard eicosanoids play an important role which has not yet been clearly determined . Bronchoconstriction and pulmonary hypertension are increased by thromboxane A2 and leukotrienes , whereas they are reduced by DB01240 . Pulmonary edema is enlarged by leukotriene , especially , LTB4 , whereas DB01240 has a protective effect . The aggregation of platelets is mediated through thromboxane A2 , PGF2 and LTB4 ; PGE1 and DB01240 counteract these reactions . LTB4 , in addition to 5-HETE , leads to the activation of inflammatory cells . Drug induced eicosanoid imbalances can be used for therapeutic interventions . Quantification of raf antisense oligonucleotide ( rafAON ) in biological matrices by LC-MS/MS to support pharmacokinetics of a liposome-entrapped rafAON formulation . An LC-MS/MS method was developed to quantify an antisense oligonucleotide against P04049 expression ( rafAON ) in monkey and mouse plasma and in mouse tissue homogenates from animals dosed with a liposome-entrapped rafAON easy-to-use formulation ( DB04973 -ETU ) intended for use as an antineoplastic agent . RafAON was extracted from mouse and monkey plasma using solid-phase extraction . Tissues were homogenized and sample cleanup was achieved by protein precipitation . RafAON and the internal standard ( IS ) were separated on a Hamilton PRP-1 column and quantified by tandem mass spectrometry using an electrospray source in negative ion mode . The total run time was 4.0 min . The peak areas of two rafAON transitions were summed and plotted against the peak area of an IS transition to generate the standard curve . In monkey plasma the linear range was 50-10,000 ng/mL , and in mouse plasma it was 25-5000 ng/mL . The lower limit of quantification was 500 ng/mL ( 10 microg/g tissue ) in heart , kidney , liver , lung and spleen homogenates , and the standard curve was linear up to 10,000 ng/mL . Accuracy , precision and stability were evaluated and found to be acceptable in all three matrices . The assay was used to support pharmacokinetics and tissue distribution studies of DB04973 -ETU in mice and monkeys . The polyglutamine neurodegenerative protein ataxin 3 regulates aggresome formation . The polyglutamine-containing neurodegenerative protein ataxin 3 ( P01008 ) has deubiquitylating activity and binds ubiquitin chains with a preference for chains of four or more ubiquitins . Here we characterize the deubiquitylating activity of P01008 in vitro and show it trims/edits K48-linked ubiquitin chains . P01008 also edits polyubiquitylated (125)I-lysozyme and decreases its degradation by proteasomes . Cellular studies show that endogenous P01008 colocalizes with aggresomes and preaggresome particles of the misfolded cystic fibrosis transmembrane regulator ( P13569 ) mutant CFTRDeltaF508 and associates with histone deacetylase 6 and dynein , proteins required for aggresome formation and transport of misfolded protein . Small interfering RNA knockdown of P01008 greatly reduces aggresomes formed by CFTRDeltaF508 , demonstrating a critical role of P01008 in this process . Wild-type P01008 restores aggresome formation ; however , P01008 with mutations in the active site or ubiquitin interacting motifs can not restore aggresome formation in P01008 knockdown cells . These same mutations decrease the association of P01008 and dynein . These data indicate that the deubiquitylating activity of P01008 and its ubiquitin interacting motifs as well play essential roles in CFTRDeltaF508 aggresome formation . DB11320 -induced P05231 and P10145 production are differentially modulated by P01579 and P05112 in human keratinocytes . It is known that large amounts of histamine are stored in mast cells located in the superficial dermis of the skin and can be released upon appropriate stimulation . However , the effects of histamine on keratinocyte function have not been well characterized . We therefore examined the capacity of histamine to modulate the production of interleukin ( IL ) -6 and P10145 by keratinocytes . We found that histamine significantly augmented the production of P05231 and P10145 in a dose- and time-dependent manner . The enhancing effects of histamine were completely inhibited by a potent H1 receptor ( P35367 ) antagonist , emedastine difumarate . Pyrilamine ( a much weaker P35367 antagonist ) and cimetidine ( an P25021 antagonist ) only partially inhibited the enhancing effects of histamine . The histamine-induced up-regulation of P05231 and P10145 production , however , was completely abrogated by a combination of DB06691 and cimetidine . The P05231 production was significantly enhanced by interferon ( IFN ) -gamma . Interestingly , P01579 and P05112 both significantly augmented the histamine-induced P05231 production . On the other hand , the production of P10145 was inhibited by P01579 , and P01579 and P05112 both completely abrogated the histamine-induced P10145 production . These results suggest that the histamine-induced P05231 production and P10145 production are differentially regulated by P01579 and P05112 . DB11320 may be an important modulator of cytokine production in epidermal milieu . Role of Bag-1 in the survival and proliferation of the cytokine-dependent lymphocyte lines , Ba/ P13726 and Nb2 . The expression and function of the newly identified Bcl-2- and P04049 - binding protein , Bag-1 , during the cytokine-regulated growth of B and T cell lines was examined . Immunoblot analysis of lysates from the interleukin-3 ( P08700 ) -dependent B cell line Ba/ P13726 , and the PRL-dependent T cell line Nb2 , revealed that variations in Bag-1 levels paralleled alterations in cellular proliferation , viability , and apoptosis induced by the presence or absence of growth factor . To test whether up-regulation of Bag-1 levels altered cellular survival and proliferation , Ba/ P13726 cells were transfected with a Bag-1 expression construct . The overexpression of Bag-1 in transfected Ba/ P13726 cells induced an P08700 -independent state . Such transfectants demonstrated sustained viability and proliferation , with minimal apoptosis , in the complete absence of exogenous P08700 . Bag-1 expression was also compared in glucocorticoid-sensitive Nb2 cells and a PRL-independent , glucocorticoid-resistant subline , SFJCD1 , during culture of these lines in dexamethasone ( DB00514 ) . Bag-1 levels were profoundly decreased by the addition of DB00514 to Nb2 cells , precedent to the onset of apoptotic cell death . In contrast , DB00514 treatment or PRL withdrawal had no effect on levels of Bag-1 within the SFJCD1 line . These findings establish that the overexpression of Bag-1 in the appropriate cellular context promotes cellular survival and growth , events that may result from the juxtaposition of this protein with mitogenic and antiapoptotic signaling pathways . DB08868 ( FTY720 ) enhances remyelination following demyelination of organotypic cerebellar slices . Remyelination , which occurs subsequent to demyelination , contributes to functional recovery and is mediated by oligodendrocyte progenitor cells ( OPCs ) that have differentiated into myelinating cells . Therapeutics that impact remyelination in the CNS could be critical determinants of long-term functional outcome in multiple sclerosis ( MS ) . DB08868 is a Q14703 receptor modulator in MS clinical trials due to systemic anti-inflammatory properties , yet may impact cells within the CNS by crossing the blood-brain barrier . Previous studies using isolated dissociated cultures indicate that neural cells express Q14703 receptors and respond to receptor engagement . Our objective was to assess the effects of fingolimod on myelin-related processes within a multicellular environment that maintains physiological cell-cell interactions , using organotypic cerebellar slice cultures . DB08868 treatment had no impact on myelin under basal conditions . DB08868 treatment subsequent to lysolecithin-induced demyelination enhanced remyelination and process extension by OPCs and mature oligodendrocytes , while increasing microglia numbers and immunoreactivity for the astrocytic marker glial fibrillary acidic protein . The number of phagocytosing microglia was not increased by fingolimod . Using Q14703 receptor specific agonists and antagonists , we determined that fingolimod-induced effects on remyelination and astrogliosis were mediated primarily through Q99500 and Q9H228 , whereas enhanced microgliosis was mediated through P21453 and Q9H228 . Taken together , these data demonstrate that fingolimod modulates multiple neuroglial cell responses , resulting in enhanced remyelination in organotypic slice cultures that maintain the complex cellular interactions of the mammalian brain . Cutting edge : suppression of T cell chemotaxis by sphingosine 1-phosphate . Murine P01730 and CD8 T cells express predominantly types 1 and 4 sphingosine 1-phosphate ( Q14703 ) G protein-coupled receptors ( designated P21453 and O95977 or previously endothelial differentiation gene-encoded 1 and 6 ) for Q14703 , which has a normal plasma concentration of 0.1-1 microM . Q14703 now is shown to enhance chemotaxis of P01730 T cells to DB00833 -21 and DB00833 -5 by up to 2.5-fold at 10 nM to 0.1 microM , whereas 0.3-3 microM Q14703 inhibits this chemotaxis by up to 70 % . Chemotaxis of Q14703 (1) , but not Q14703 (4) , transfectants to P09341 and P02776 was similarly affected by Q14703 . Activation of P01730 T cells , which decreases Q14703 receptor expression , suppressed effects of Q14703 on chemotaxis . Pretreatment of labeled P01730 T cells with Q14703 before reintroduction into mice inhibited by a maximum of 75 % their migration into chemokine-challenged s.c. air pouches . The Q14703 - Q14703 (1) receptor axis thus controls recruitment of naive T cells by maintaining their response threshold to diverse lymphotactic factors . A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins . A surface plasmon resonance-based solution affinity assay is described for measuring the K(d) of binding of heparin/heparan sulfate-binding proteins with a variety of ligands . The assay involves the passage of a pre-equilibrated solution of protein and ligand over a sensor chip onto which heparin has been immobilised . DB01109 sensor chips prepared by four different methods , including biotin-streptavidin affinity capture and direct covalent attachment to the chip surface , were successfully used in the assay and gave similar K(d) values . The assay is applicable to a wide variety of heparin/HS-binding proteins of diverse structure and function ( e.g. , P05230 , P09038 , P15692 , P10145 , P80075 , P01008 , P02776 ) and to ligands of varying molecular weight and degree of sulfation ( e.g. , heparin , DB05808 , sucrose octasulfate , naphthalene trisulfonate ) and is thus well suited for the rapid screening of ligands in drug discovery applications . 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 . [ 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 .

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interacts_with DB08899?

multiple_choice

Endotoxin-induced liver damage in rats is minimized by beta 2-adrenoceptor stimulation . OBJECTIVE AND DESIGN : To investigate the effects of beta(2)-adrenoceptor ( beta(2)-AR ) stimulation on endotoxin-induced liver damage and systemic cytokine levels in rats . SUBJECTS : Standard male Wistar rats . TREATMENT : A disease-model of lipopolysaccharide ( LPS ) -induced acute systemic inflammation was used . The beta(2)-selective AR agonist clenbuterol was administered before , during , and after LPS-challenge to investigate its effects on the acute inflammatory response and associated liver-failure . METHODS : The following parameters have been measured in plasma : P01375 alpha , P01584 , P05231 , P22301 , Q9NRA2 , ALT , and Bilirubin . Liver histological examination was performed to look for changes in tissue morphology . RESULTS : Administration of clenbuterol ( p.o. ) one hour before , or intravenous at the same time as LPS-challenge resulted in a marked reduction of plasma levels of P01375 alpha , P01584 , and P05231 . A change both in plasma-level and in time-concentration profile of the anti-inflammatory cytokine P22301 was found . DB01407 minimized LPS-induced liver damage , as represented by significantly lowered concentrations of several parameters for liver-failure ( Q9NRA2 , ALT , Bilirubin ) , and improved hepatic tissue morphology . DB01407 administration after LPS challenge failed to inhibit P01375 alpha-release but reduced liver-damage . Simultaneous use of the beta(2)-AR antagonist propranolol augmented LPS-induced liver failure , suggesting a role of endogenous adrenoceptor-agonists in prevention of organ-failure during systemic inflammation . CONCLUSIONS : The results indicate that a selective beta(2)-AR agonist might be used as an additional therapeutic agent in the clinic for the treatment of ( acute ) systemic inflammatory disorders in order to reduce or prevent subsequent liver failure . 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 . Differential targeting of brain stress circuits with a selective glucocorticoid receptor modulator . P04150 ( GR ) antagonism may be of considerable therapeutic value in stress-related psychopathology such as depression . However , blockade of all GR-dependent processes in the brain will lead to unnecessary and even counteractive effects , such as elevated endogenous cortisol levels . Selective GR modulators are ligands that can act both as agonist and as antagonist and may be used to separate beneficial from harmful treatment effects . We have discovered that the high-affinity GR ligand C108297 is a selective modulator in the rat brain . We first demonstrate that C108297 induces a unique interaction profile between GR and its downstream effector molecules , the nuclear receptor coregulators , compared with the full agonist dexamethasone and the antagonist DB00834 ( mifepristone ) . C108297 displays partial agonistic activity for the suppression of hypothalamic corticotropin-releasing hormone ( P06850 ) gene expression and potently enhances GR-dependent memory consolidation of training on an inhibitory avoidance task . In contrast , it lacks agonistic effects on the expression of P06850 in the central amygdala and antagonizes GR-mediated reduction in hippocampal neurogenesis after chronic corticosterone exposure . Importantly , the compound does not lead to disinhibition of the hypothalamus-pituitary-adrenal axis . Thus , C108297 represents a class of ligands that has the potential to more selectively abrogate pathogenic GR-dependent processes in the brain , while retaining beneficial aspects of GR signaling . Androgen-induced coactivator Q6PL18 mediates specific androgen receptor signaling in prostate cancer . P10275 ( AR ) plays a pivotal role in prostate cancer , primarily by regulating different gene expression programs elicited by androgen , which is important for cancer cell proliferation , survival , and differentiation . It is believed that the transcriptional function of AR is mediated largely by distinct nuclear coregulators . We report here the identification of Q6PL18 ( also known as Q6PL18 ) , a new member of the AAA+ ATPase family proteins , as a novel AR coactivator . Q6PL18 interacts directly with AR and enhances its transcriptional activity , and is required for androgen-stimulated expression of a specific subgroup of genes including P08069 , Q9Y4H2 , O00141 , and survivin . Upon androgen stimulation , Q6PL18 together with AR is recruited to the specific AR target genes . Suppression of Q6PL18 expression strongly inhibited the proliferation of androgen-responsive or androgen-independent , AR-positive prostate cancer cells and caused a significant increase of cellular apoptosis . Strikingly , the Q6PL18 gene itself , located at chromosome 8q24 , is highly induced by androgen in androgen-dependent prostate cancer cells and xenograft tumors . Although Q6PL18 is hardly detected in normal human prostate tissue , high levels of Q6PL18 are found in hormone-independent prostate cancer cell lines , xenograft tumor , and a subset of prostate cancers with high Gleason scores . Together , these findings suggest that Q6PL18 plays an important role in prostate cancer by mediating specific AR functions in cancer cell survival and proliferation . The possession of ATPase and bromodomain by Q6PL18 makes it an attractive target for the development of therapeutics for the disease . 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 . Parathyroid hormone activation of matrix metalloproteinase-13 transcription requires the histone acetyltransferase activity of p300 and Q92831 and p300-dependent acetylation of Q92831 . Parathyroid hormone ( PTH ) regulates the transcription of many genes involved in bone remodeling in osteoblasts . One of these genes is matrix metalloproteinase-13 ( P45452 ) , which is involved in bone remodeling and early stages of endochondral bone formation . We have previously shown that Mmp-13 gene expression is highly induced by PTH treatment in osteoblastic UMR 106-01 cells , as well as primary osteoblasts . Here , we show that p300/CBP-associated factor ( Q92831 ) , in addition to p300 and Runx2 , is required for PTH activation of Mmp-13 transcription . Q92831 was increasingly recruited to the P45452 proximal promoter region after PTH treatment , and this was associated with an increase in RNA polymerase II recruitment and histone acetylation . In addition , PTH treatment increased the acetylation of Q92831 , a process that required p300 . Knockdown of Q92831 , p300 , or Runx2 by siRNA decreased Mmp-13 mRNA expression after PTH treatment in both UMR 106-01 cells and primary osteoblasts . We found that there is a mutual dependence between p300 and Q92831 to be recruited to the Mmp-13 promoter after PTH treatment . In promoter-reporter assays , p300 and Q92831 had an additive effect on PTH stimulation of P45452 promoter activity , and this required their histone acetyltransferase activity . Our findings demonstrate that Q92831 acts downstream of PTH signaling as a transcriptional coactivator that is required for PTH stimulation of P45452 transcription . Q92831 cooperates with p300 and Runx2 to mediate PTH activation of P45452 transcription . The regulation of Q05469 and P06858 expression by DB02901 and flutamide in human subcutaneous adipose tissue . Clinical observations suggest a role for testosterone in the accumulation of central adiposity and with an associated increased risk of disease . To date , no human study has analysed the role of dihydrotestosterone ( DB02901 ) on adipose tissue mass regulation in vitro . This study investigated the role of DB02901 and androgen receptors ( AR ) in the regulation of lipolysis and lipogenesis by examining the key enzymes hormone sensitive lipase ( Q05469 ) and lipoprotein lipase ( P06858 ) respectively . Isolated abdominal subcutaneous adipocytes ( Scad ) ( n = 15 ) were treated with either DB02901 ( 10(-7)-10(-9) m ) , an antiandrogen , flutamide ( P17948 : 10(-7)-10(-9) m ) or a combination of DB02901 ( 10(-7)-10(-9) m ) with P17948 ( 10(-8) m ) . Relative protein expression of Q05469 , P06858 and AR was determined . In Scad , DB02901 inhibited Q05469 expression maximally at 10(-9) m ( 0.7 +/- 0.4** ; p < 0.01** ) compared with control ( control : 1.0 +/- ( s.e.m. ) 0.0 ) , whereas P06858 protein expression was stimulated at DHT10(-9) m ( 2.22 +/- 0.48* ; p < 0.05* ) . DB04077 release assay results correlated with Q05469 expression data . P06858 expression was reduced at all doses with combinations of DB02901 + P17948 compared with DB02901 alone . P10275 expression studies showed an inverse correlation with DB02901 , whereas DB02901 + P17948 reduced AR expression . These studies indicate that DB02901 may alter Q05469 and P06858 expression , whereas only P06858 expression appears mediated by AR . These findings suggest a physiological role for DB02901 in the control of adipose tissue mass in women , and indicate that androgens may also play an important role in regulating lipid metabolism . 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 . 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 . 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 . Hormonal regulation of human trophoblast differentiation : a possible role for 17beta-estradiol and DB00644 . We have examined the role of 17beta-estradiol and gonadotropin releasing hormone ( DB00644 ) in the regulation of functional differentiation in human trophoblasts . In contrast to its recognized functions as a proliferation-promoting hormone in a variety of cell types , we found that 17beta-estradiol induced terminal differentiation in human trophoblastic cells , and that this event was estrogen-receptor-mediated . This process involved a loss in expression of Cyclins A2 and E , and a coincident increase in p27(Kip1) . The anti-proliferative effects of 17beta-estradiol were annulled by specific transforming growth factor-beta 1 ( TGFbeta1 ) -neutralizing antibody , suggesting that 17beta-estradiol may mediate its growth-inhibitory actions , through TGFbeta1 activity . Following exposure to DB06719 , cultured human trophoblastic cells stopped proliferating and formed functionally mature syncytiotrophoblasts . This differentiation event , that involved a drastic loss in expression of proliferating-cell-nuclear-antigen , could be blocked by DB00050 , suggesting the involvement of functional DB00644 receptors . Preliminary studies on the characterization of the human placental P30968 , indicate the presence of multiple receptor isoforms across human gestation . 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 . 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 . miR-17-5p targets the p300/CBP-associated factor and modulates androgen receptor transcriptional activity in cultured prostate cancer cells . BACKGROUND : P10275 ( AR ) signalling is critical to the initiation and progression of prostate cancer ( PCa ) . Transcriptional activity of AR involves chromatin recruitment of co-activators , including the p300/CBP-associated factor ( Q92831 ) . Distinct miRNA expression profiles have been identified in PCa cells during the development and progression of the disease . Whether miRNAs regulate Q92831 expression in PCa cells to regulate AR transcriptional activity is still unclear . METHODS : Expression of Q92831 was investigated in several PCa cell lines by qRT-PCR , Western blot , and immunocytochemistry . The effects of Q92831 expression on AR-regulated transcriptional activity and cell growth in PCa cells were determined by chromatin immunoprecipitation , reporter gene construct analysis , and MTS assay . Targeting of Q92831 by miR-17-5p was evaluated using the luciferase reporter assay . RESULTS : Q92831 was upregulated in several PCa cell lines . Upregulation of Q92831 promoted AR transcriptional activation and cell growth in cultured PCa cells . Expression of Q92831 in PCa cells was associated with the downregulation of miR-17-5p . Targeting of the 3'-untranslated region of Q92831 mRNA by miR-17-5p caused translational suppression and RNA degradation , and , consequently , modulation of AR transcriptional activity in PCa cells . CONCLUSIONS : Q92831 is upregulated in cultured PCa cells , and upregulation of Q92831 is associated with the downregulation of miR-17-5p . Targeting of Q92831 by miR-17-5p modulates AR transcriptional activity and cell growth in cultured PCa cells . DB00644 -mediated phosphorylation of estrogen receptor-alpha contributes to fosB expression in mouse gonadotrophs . Estrogen receptors ( ERs ) are activated by their ligands as well as signaling pathways that alter ER phosphorylation in response to peptide hormones and growth factors . In pituitary gonadotrophs , GnRHs act via the type I P30968 ( GnRHR ) . Both DB00644 subtypes ( DB00644 and -II ) activate an estrogen response element ( ERE ) -driven luciferase reporter gene in LbetaT2 mouse pituitary cells , and DB00644 is most potent in this regard . Moreover , antide ( a DB00644 antagonist ) and a GnRHR small interfering RNA ( siRNA ) abrogate this effect , whereas an ERalpha antagonist ( DB00947 ) does not . The ERalpha in LbetaT2 cells is phosphorylated at DB00133 (118) in the nucleus and at DB00133 (167) in both nucleus and cytoplasm after DB00644 treatments and coincided with increased ERalpha binding to its coactivator , the p300/ DB02527 response element-binding protein-associated factor ( Q92831 ) . Moreover , siRNA-mediated knockdown of Q92831 levels attenuated DB00644 -induced ERE-luciferase transactivation in these cells . Most importantly , both DB00644 subtypes robustly up-regulated expression of the immediate early response gene , fosB , whereas cotreatment with ERalpha siRNA or Q92831 siRNA attenuated this effect . This appears to occur at the transcriptional level because corecruitment of ERalpha and Q92831 to an ERE within the endogenous fosB promoter was increased by DB00644 treatments , as shown by chromatin immunoprecipitation assays . These data demonstrate that DB00644 -mediated phosphorylation of ERalpha in mouse LbetaT2 pituitary cells results in its rapid association with Q92831 and the transcriptional activation of fosB , and we demonstrate that this in turn likely activates other genes in pituitary cells including the DB00094 beta-subunit gene . 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 . Lymphocyte depletion and repopulation in peripheral blood and small intestine of cynomolgus monkeys after alemtuzumab treatment . BACKGROUND : DB00087 has been used as an induction agent in organ transplantation over 10 years , but the effect of alemtuzumab on lymphocytes in small intestine was not clear . We investigate lymphocyte depletion and repopulation phenomena both in peripheral blood and small intestine of cynomolgus monkeys , to assess the model using in preclinical transplantation . MATERIALS AND METHODS : Monkeys without P31358 antigen on erythrocytes were selected . Lymphocyte depletion and repopulation was documented by flow cytometry . Sections of ileum were obtained for isolation of intestinal intraepithelial lymphocytes ( IEL ) and lamina propria lymphocytes ( P06858 ) , and also for immunofluorescence examination . RESULTS : Powerful depletion of lymphocytes ( > 80 % ) from blood followed by gradual repopulation was observed . P11836 (+) B cells , CD8(+) T cells , P01730 (+) T cells returned to pretreatment levels by d 21 , 35 , 56 . IEL , P06858 reduced by 70 % , 72 % on d 9 , recovered to 59 % , 57 % of pretreatment levels by d 35 , and were completed by d 56 . Depletion and repopulation of IEL and P06858 were confirmed by immunofluorescence . CONCLUSIONS : Depletion of lymphocytes in peripheral blood was less powerful and repopulation occurred faster than in patients . The lymphocyte depletion and repopulation occurred in small intestine . This model can be used in preclinical transplantation . 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 . 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 . Histone deacetylase inhibition mediates urocortin-induced antiproliferation and neuronal differentiation in neural stem cells . During cortical development , cell proliferation and cell cycle exit are carefully regulated to ensure that the appropriate numbers of cells are produced . P55089 ( P55089 ) is a member of the corticotrophin releasing hormone ( P06850 ) family of neuropeptides that regulates stress responses . P55089 is widely distributed in adult rat brain . However , the expression and function of P55089 in embryonic brain is , as yet , unclear . Here , we show that P55089 is endogenously expressed in proliferative zones of the developing cerebral cortex and its receptors are exhibited in neural stem cells ( NSCs ) , thus implicating the neuropeptide in cell cycle regulation . Treatment of cultured NSCs or organotypic slice cultures with P55089 markedly reduced cell proliferation . Furthermore , blocking of endogenous P55089 /CRHRs system either by treatment with CRHRs antagonists or by neutralization of secreted P55089 with anti- P55089 antibody increased NSCs proliferation . Cell cycle kinetics analysis demonstrated that P55089 lengthened the total cell cycle duration via increasing the P55008 phase and accelerated cell cycle exit . P55089 directly inhibited the histone deacetylase ( HDAC ) activity and induced a robust increase in histone H3 acetylation levels . Using pharmacological and RNA interference approaches , we further demonstrated that antiproliferative action of P55089 appeared to be mediated through a HDAC inhibition-induced P38936 upregulation . Moreover , P55089 treatment in vitro and in vivo led to an increase in neuronal differentiation of NSCs . These findings suggest that P55089 might contribute to regulate NSCs proliferation and differentiation during cortical neurogenesis .

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interacts_with DB00762?

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 . d- DB00574 - and d-norfenfluramine-induced hypophagia : differential mechanisms and involvement of postsynaptic 5-HT receptors . Severe depletion of 5-hydroxytryptamine ( 5-HT ) by para-chlorophenylalanine ( pCPA , 150 mg/kg per day x3 ) did not alter the hypophagic effect of d-fenfluramine ( 1-3 mg/kg i.p. ) 1 h after food presentation in 24-h food-deprived rats , and moderately and comparably increased the hypophagic effects of its metabolite , d-norfenfluramine ( 0.35-1.0 mg/kg i.p. ) , and of the P28335 receptor agonist , 1-(3-chlorophenyl)piperazine ( mCPP ; 1.5 , 2.0 mg/kg i.p. ) . Chronic treatment with mCPP ( 2.5 mg/kg i.p. x 14 ) attenuated the hypophagia induced by d-norfenfluramine ( 1 , 1.5 mg/kg ) but not d-fenfluramine ( 1 , 3 mg/kg ) . 1-(1-Naphthyl)piperazine ( 3 , 8 mumol/kg s.c. ) , which has greater affinity for P28335 than for 5-HT2 receptors , had no effect on the hypophagia induced by d-fenfluramine ( 1.25 , 2.0 mg/kg ) , but 1.3 and 3 mumol/kg 1-(1-naphthyl)piperazine largely and comparably attenuated the substantial hypophagic effect of d-norfenfluramine ( 0.75 mg/kg ) . The essentially complete hypophagic action of d-norfenfluramine ( 1.25 mg/kg ) was inhibited by 1-(1-naphthyl)piperazine with ID50 = 2.13 mumol/kg . Ketanserin , which binds more weakly than 1-(1-naphthyl)piperazine to P28335 receptors and more strongly to 5-HT2 receptors , attenuated weaker but not stronger hypophagic effects of d-fenfluramine ( 1.25 , 2.0 mg/kg ) when given at high dosage ( 8 , 16 mumol/kg s.c. ) . Ketanserin ( 16 mumol/kg ) also weakly attenuated the hypophagia due to d-norfenfluramine ( 0.75 mg/kg ) , but not the essentially complete hypophagia due to d-norfenfluramine ( 1.25 mg/kg ) . ( ABSTRACT TRUNCATED AT 250 WORDS ) Dexamethasone potentiates serotonin-2 receptor-mediated intracellular Ca2+ mobilization in P13671 glioma cells . Serotonin ( 5-hydroxytryptamine ; 5-HT ) caused a transient increase in intracellular Ca2+ in C6BU-1 glioma cells in a concentration-dependent manner ; half maximally at 73 nM . The 5-HT2 agonist 1-(4-iodo-2,5-dimethoxyphenyl)-2- aminopropane also increased the levels of intracellular Ca2+ , whereas the P28335 agonist 1-(3-chlorophenyl)piperazine and P08908 agonist 8-hydroxy-2- (di-n-propylamino)tetralin were completely ineffective . Ketanserin and spiperone blocked the response to 5-HT at a nanomolar concentration , but the 5- Q9H205 antagonist MDL 72222 had no effect on it . Thus 5-HT2 receptors are responsible for activating Ca2+ mobilization in P13671 glioma cells . Treatment of P13671 glioma cells with dexamethasone potentiated the ability of 5-HT to cause intracellular Ca2+ mobilization in both a dose- and time-dependent manner . The dose-response curve for 5-HT was shifted 9-fold to the left compared to controls , and the Vmax value was also significantly enhanced . This enhanced Ca2+ mobilization was completely inhibited by ketanserin dose-dependently . In addition , the treatment with dexamethasone enhanced fluoride-activated Ca2+ mobilization , suggesting that the enhanced GTP binding protein function is one of the mechanisms responsible for the enhancement of the 5-HT response induced by dexamethasone treatment . This enhancement of agonist activity was mediated by the type II glucocorticoid receptor ( GR ) since RU 38486 , an inhibitor of the type II GR , antagonized the dexamethasone-induced enhancement . 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 . Exendin-4 alleviates angiotensin II-induced senescence in vascular smooth muscle cells by inhibiting Rac1 activation via a DB02527 /PKA-dependent pathway . Vascular aging has been implicated in the progression of diabetes and age-related cardiovascular disorders . Glucagon-like peptide-1 ( P0C6A0 ) is an incretin hormone capable of cytoprotective actions in addition to its glucose-lowering effect . The present study was undertaken to examine whether Exendin-4 , a specific ligand for the P43220 , could prevent angiotensin ( P03950 ) II-induced premature senescence in vascular smooth muscle cells ( VSMCs ) and to determine the underlying mechanism involved . Senescence-associated β-galactosidase ( SA β-gal ) assay showed that P03950 II induced premature senescence of VSMCs . Pretreatment with Exendin-4 significantly attenuated P03950 II-induced generation of H2O2 and the subsequent VSMC senescence . These effects were , however , reversed in the presence of exendin fragment 9-39 , a P43220 antagonist , or PKI14-22 . Moreover , a marked increase in the levels of p53 and P38936 induced by P03950 II was blunted by the treatment with Exendin-4 . Nevertheless , Exendin-4 failed to decrease P03950 II-induced expression of NAD(P)H oxidase 1 ( Nox1 ) , NAD(P)H oxidase 4 ( Nox4 ) , O75935 (phox) , or p47(phox) in VSMCs . Mechanistically , Exendin-4 blocked P03950 II-induced Rac1 activation through the DB02527 /PKA signaling cascade . Specifically , NSC23766 , a Rac1 inhibitor , abrogated the suppressive effects of Exendin-4 on P03950 II-induced premature senescence and H2O2 generation , respectively . Thus Exendin-4 confers resistance to P03950 II-induced superoxide anion generation from NAD(P)H oxidase and the resultant VSMC senescence by inhibiting Rac1 activation via a DB02527 /PKA-dependent pathway . These findings demonstrate that P0C6A0 as well as its analogs ( P0C6A0 -related reagents ) may hold therapeutic potential in the treatment of diabetes with cardiovascular disease . Familial nonmedullary thyroid cancer : a review of the genetics . OBJECTIVE : Thyroid cancer , the commonest of endocrine malignancies , continues to increase in incidence with over 19,000 new cases diagnosed in the European Union per year . Although nonmedullary thyroid cancer ( NMTC ) is mostly sporadic , evidence for a familial form , which is not associated with other Mendelian cancer syndromes ( e.g. , familial adenomatous polyposis and Cowden 's syndrome ) , is well documented and thought to cause more aggressive disease . Just over a decade ago , the search for a genetic susceptibility locus for familial NMTC ( FNMTC ) began . This review details the genetic studies conducted thus far in the search for potential genes for FNMTC . DESIGN : An electronic PubMed search was performed from the English literature for genetics of FNMTC and genetics of familial papillary thyroid carcinoma ( subdivision of FNMTC ) . The references from the selected papers were reviewed to identify further studies not found in the original search criteria . MAIN OUTCOME : Six potential regions for harboring an FNMTC gene have been identified : MNG1 ( 14q32 ) , TCO ( 19p13.2 ) , fPTC/ Q9H6Q4 ( 1q21 ) , NMTC1 ( 2q21 ) , FTEN ( 8p23.1- O75935 ) , and the telomere-telomerase complex . Important genes reported to have been excluded are P07949 , P04629 , MET , P25054 , P60484 , and P16473 . CONCLUSION : The genetics of FNMTC is an exciting field in medical research that has the potential to permit individualized management of thyroid cancer . Studies thus far have been on small family groups using varying criteria for the diagnosis of FNMTC . Results have been contradictory and further large-scale genetic studies utilizing emerging molecular screening tests are warranted to elucidate the underlying genetic basis of FNMTC . 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 . Enhanced sensitivity to irinotecan by Cdk1 inhibition in the p53-deficient HT29 human colon cancer cell line . Mutations in the tumor-suppressor gene p53 have been associated with advanced colorectal cancer ( CRC ) . DB00762 ( CPT-11 ) , a P11387 inhibitor , has been recently incorporated to the adjuvant therapy . Since the DNA-damage checkpoint depends on p53 activation , the status of p53 might critically influence the response to CPT-11 . We analysed the sensitivity to CPT-11 in the human colon cancer cell line HT29 ( mut p53 ) and its wild-type (wt)-p53 stably transfected subclone HT29-A4 . Cell-cycle analysis in synchronised cells demonstrated the activation of transfected wt-p53 and a P38936 ( P38936 /CIP1)-dependent cell-cycle blockage in the S phase . Activated wt-p53 increased apoptosis and enhanced sensitivity to CPT-11 . In p53-deficient cells , cDNA-macroarray analysis and western blotting showed an accumulation of the cyclin-dependent kinase (cdk)1/cyclin B complex . Subsequent p53-independent activation of the cdk-inhibitor ( cdk-I ) P38936 ( P38936 /CIP1) prevented cell-cycle progression . Cdk1 induction was exploited in vivo to improve the sensitivity to CPT-11 by additional treatment with the cdk-I P99999 -202 . We demonstrate a gain of sensitivity to CPT-11 in a p53-mutated colon cancer model either by restoring wild-type p53 function or by sequential treatment with cdk-Is . Considering that mutations in p53 are among the most common genetic alterations in CRC , a therapeutic approach specifically targeting p53-deficient tumors could greatly improve the treatment outcomes . 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 . On-target effects of P43220 agonists on thyroid C-cells in rats and mice . Glucagon-like peptide-1 is an incretin hormone from the gastrointestinal tract , which enhances insulin secretion , slows gastric emptying , and reduces food intake . P43220 agonists are being developed for Type 2 diabetes mellitus . P0C6A0 is rapidly degraded by serum dipeptidyl peptidase IV , so analogues with a prolonged serum half-life are used clinically . Exenatide was the first P0C6A0 agonist approved and is a synthetic version of exendin-4 derived from the Gila monster . DB06655 was approved for clinical use in 2010 . P43220 agonists have been shown to increase calcitonin secretion and stimulate C-cell hyperplasia and neoplasia in rats and mice of both sexes . Rat C-cells are more sensitive to the effects of P0C6A0 agonists than mice . The effects of P0C6A0 agonists on C-cell proliferation or neoplasia have not been documented in nonhuman primates or humans . The proliferative C-cell effects may be rodent-specific . P0C6A0 receptors have been demonstrated on normal rodent C-cells , but are either not present or occur in low numbers on C-cells of nonhuman primates and humans . Hyperplasia and neoplasia of C-cells in rodents treated with P0C6A0 agonists represent a unique example of an on-target species-specific effect that may not have relevance to humans . 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 . Effect of oncostatin M on uridine diphosphate-5'-glucuronosyltransferase 1A1 through cross talk with constitutive androstane receptor . Hyperbilirubinemia remains a common condition in neonates . The constitutive androstane receptor ( CAR ) is an orphan nuclear receptor that has been shown to participate in the activation of the uridine diphosphate-5'-glucuronosyltransferase 1A1 ( P22309 ) gene , which plays an important role in bilirubin clearance . Oncostatin M ( P13725 ) , a member of the P05231 family , is involved in the maturation of fetal hepatocytes . We have demonstrated that low P13725 levels are a potential indicator of neonatal jaundice and the need for phototherapy . In this study we examined the effects of P13725 on CAR-mediated signaling to investigate its potential role in neonatal jaundice via the CAR- P22309 pathway . We observed that P13725 positively augmented the CAR and P22309 expressions and CAR-mediated signaling in vivo and in vitro , through cross talk between the nuclear CAR receptor and the plasma membrane P13725 receptor , via the MAPK cascade . These data suggest that P13725 might play a role in bilirubin metabolism via the CAR- P22309 pathway . Antihistamine effects on prefrontal cortex activity during working memory process in preschool children : a near-infrared spectroscopy ( NIRS ) study . P35367 antagonists ( antihistamines ) are widely used for the treatment of allergic disorders in young children . This study examined the effects of antihistamine on prefrontal cortex activity in preschool children using near-infrared spectroscopy ( NIRS ) , an emerging brain-imaging method suitable for psychological experiments , especially in young children . We examined the changes of oxygenated hemoglobin concentration in the prefrontal cortex while children performed a spatial working memory task , 3h after taking a first-generation antihistamine ( ketotifen ) , second-generation antihistamine ( epinastine ) , or placebo . Fifteen healthy preschool children ( mean age , 5.5 years ) participated . DB00920 significantly impaired behavioral performance and cortical activation at the lateral prefrontal cortex in the working memory task , compared with epinastine and placebo . There were no sedative effects on neural response or behavioral performance after epinastine administration . This paper demonstrates for the first time differential sedation effects of first- and second-generation antihistamines on brain hemodynamic response in young children . Also discussed is the utility of the NIRS technique in neuropsychopharmacological studies of children . 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 . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . DB04725 , a dual P36551 /5- P28300 inhibitor , induces apoptosis in HCA-7 colon cancer cells through the mitochondrial pathway independently from its ability to affect the arachidonic acid cascade . Nowadays , no data are available concerning the potential use of dual cyclooxygenase ( P36551 ) / P09917 ( P28300 ) inhibitors as anticancer agents in colon cancer treatment . Here , we report , for the first time , that the dual P36551 /5- P28300 inhibitor licofelone triggers apoptosis in a dose- and time-dependent manner in HCA-7 colon cancer cells . Induction of apoptosis was related to the recruitment of the intrinsic mitochondrial apoptotic pathway , as shown by loss in mitochondrial membrane potential , cytochrome c release , caspase-9 and 3 activation and poly-(ADP-ribose)polymerase-1 cleavage . Moreover , licofelone induced the cleavage of the full-length P38936 (Bax) into p18(Bax) , a more potent inducer of the apoptotic process than the uncleaved form . Pre-treatment of HCA-7 cells with the pan-caspase inhibitor z-VAD-fmk significantly blocked licofelone-induced apoptosis , confirming that this process occurred primarily in a caspase-dependent pathway . We also present evidences that licofelone was able to affect the arachidonic acid ( AA ) cascade , as it blocked the activity of 5- P28300 and P36551 enzymes , and it induced , through the phosphorylation of cytoplasmic phospholipase A(2) ( cPLA(2) ) , the release of unesterified AA from HCA-7 membrane phospholipids . However , apoptosis induction was not related to the ability of licofelone to affect the AA cascade , since neither exogenous prostaglandin E(2) and leukotriene B(4) addition , nor pharmacological inhibition of cPLA(2) , was able to rescue HCA-7 cells from apoptosis . Even if further studies are needed to clarify the mechanism of licofelone-induced apoptosis , this study suggests that this drug , as well as similar dual P36551 /5- P28300 inhibitors , may represent a novel and promising approach in colon cancer treatment . 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 . Novel and emerging drugs for acute myeloid leukemia : pharmacology and therapeutic activity . For the last twenty years , significant progress in Molecular and Cellular Biology has resulted in a better characterization and understanding of the biology and prognosis of acute myeloid leukemia ( AML ) . These achievements have provided new opportunities for the development of innovative , more effective therapies . Novel agents potentially useful in the treatment of patients with AML include new formulations of established drugs , newer nucleoside analogs , molecular target drugs , monoclonal antibodies and other agents . Three newer nucleoside analogs , clofarabine , troxacitabine and sapacitabine have been recently investigated in patients with AML . Two methylation inhibitors , 5-azacyticline and decitabine are pyrimidine nucleoside analogs of cytidine which can be incorporated into RNA and/or DNA . Lower doses of these agents are active in AML and have been extensively investigated , especially in secondary AML and AML in elderly patients . DB04960 and lonafarnib are orally available farnesyltransferase inhibitors with in vitro and in vivo activity against AML . In recent years , P36888 inhibitors , lestaurinib , DB05465 and PKC 412 have been developed and tested in AML . The preclinical observations and clinical studies indicate that P36888 inhibitors are promising agents in the treatment of P36888 mutated AML patients , especially when used in combinations with chemotherapy . Several newer MDR inhibitors , including valspodar ( PSC-833 ) and zosuquidar trihydrochloride have been also tested for the treatment of relapsed AML . This article reviews the various classes of AML targets and drugs that are under early phase clinical evaluation , especially those that are likely to enter clinical practice in the near future . Protective effects of heparin on endothelial cells in sepsis . This study aims to observe the protective effects of heparin on endothelial cells in sepsis and explore the involved signal pathway regulated by heparin . Methods Human vascular endothelial cells were treated by TNFα in vitro to simulate the inflammatory environment when sepsis occurred . They were intervened by heparin and the expression levels of soluble thrombomodulin ( sTM ) and serum activated protein C ( P25054 ) were detected by ELISA , the regulatory mechanism of heparin improving vascular endothelial cells injury induced by TNFα was detected by Western Blotting method , the methylation of histone in the gene promoter region of endothelial nitric oxide synthase ( P29474 ) and monocyte chemotactic protein-1 ( P13500 ) were detected using chromatin immunoprecipitation method . Results DB01109 could inhibit the secretion of sTM and P25054 protein and the expression of P13500 gene which involved in NF-κB signal pathway . Conclusions DB01109 could protect vascular endothelial cells from injury induced by TNFα and sepsis , the mechanisms were related with the effects of heparin on the histone methylation of promoter region and the regulation of heparin on the MAPK and NF-κB signal pathways . These results provide a theoretical basis for the application of heparin in the prevention and treatment of vascular disease related with sepsis . Rapid generation of antigen-presenting cells from leukaemic blasts in acute myeloid leukaemia . The ability of acute myeloid leukaemia ( AML ) cells to acquire dendritic cell ( DC ) -like characteristics in vitro with a rapid culture method based either on the phorbol ester PMA or calcium ionophores has been studied in comparison to conventional AML-DC cultures with the cytokines granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) , tumour necrosis factor-alpha ( P01375 ) , interleukin-3 ( P08700 ) , P21583 , P36888 -L and P05112 . In all AML patients , antigen-presenting cells ( P25054 ) could be generated from leukaemic cells in 2 days by incubation with PMA or calcium ionophore ( A23187 or ionomycin ) in the presence as well as in the absence of P05112 . In 30 out of 36 patients P25054 could be generated after 2 weeks of culture in cytokine-enriched medium . AML- P25054 cultured with PMA or calcium ionophores immunophenotypically and functionally were at a more mature stage than those cultured in cytokine-enriched medium . The most mature P25054 were generated by calcium ionophore A23187 plus P05112 , as evidenced by the higher expression of P25942 , P33681 , P42081 and HLA-DR . Autologous T cell mediated cytotoxicity towards AML blast cells in vitro was observed in 2 cases tested . The persistence of cytogenetic abnormalities confirmed the leukaemic origin of the AML- P25054 . The generation of AML- P25054 was possible from freshly isolated as well as cryopreserved material . Our data show that generation of sufficient AML- P25054 by A23187 plus P05112 is feasible , for vaccination purposes , in approximately 70 % of AML specimens , offering a time-saving and cost-effective approach in preparing anti-leukaemia vaccines .

MH_train_1515

MH_train_1515

MH_train_1515

interacts_with DB01418?

multiple_choice

Characterization of 107 genomic DNA reference materials for P10635 , P33261 , P11712 , Q9BQB6 , and P22309 : a GeT-RM and Association for Molecular Pathology collaborative project . Pharmacogenetic testing is becoming more common ; however , very few quality control and other reference materials that cover alleles commonly included in such assays are currently available . To address these needs , the Centers for Disease Control and Prevention 's Genetic Testing Reference Material Coordination Program , in collaboration with members of the pharmacogenetic testing community and the Coriell Cell Repositories , have characterized a panel of 107 genomic DNA reference materials for five loci ( P10635 , P33261 , P11712 , Q9BQB6 , and P22309 ) that are commonly included in pharmacogenetic testing panels and proficiency testing surveys . Genomic DNA from publicly available cell lines was sent to volunteer laboratories for genotyping . Each sample was tested in three to six laboratories using a variety of commercially available or laboratory-developed platforms . The results were consistent among laboratories , with differences in allele assignments largely related to the manufacturer 's assay design and variable nomenclature , especially for P10635 . The alleles included in the assay platforms varied , but most were identified in the set of 107 DNA samples . Nine additional pharmacogenetic loci ( P78329 , P07099 , P08183 , HLAB , Q6ZMV9 , P08684 , P20815 , P51580 , and Q12882 ) were also tested . These samples are publicly available from Coriell and will be useful for quality assurance , proficiency testing , test development , and research . The human SWI/SNF complex associates with Q01196 to control transcription of hematopoietic target genes . The acute myeloid leukemia 1 ( Q01196 , Q01196 ) transcription factor is a key regulator of hematopoietic differentiation that forms multi-protein complexes with co-regulatory proteins . These complexes are assembled at target gene promoters in nuclear microenvironments to mediate phenotypic gene expression and chromatin-related epigenetic modifications . Here , immunofluorescence microscopy and biochemical assays are used to show that Q01196 associates with the human DB00171 -dependent SWI/SNF chromatin remodeling complex . The SWI/SNF subunits P51532 and Q12824 bind in vivo to Q01196 target gene promoters ( e.g. , P04141 , P08700 , P09603 -R , MIP , and P38936 ) . These interactions correlate with histone modifications characteristic of active chromatin , including acetylated H4 and dimethylated H3 lysine 4 . Downregulation of Q01196 by RNA interference diminishes the binding of P51532 and Q12824 at selected target genes . Taken together , our findings indicate that Q01196 interacts with the human SWI/SNF complex to control hematopoietic-specific gene expression . 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 . Inhibition of natural killer cells protects the liver against acute injury in the absence of glycine N-methyltransferase . Q14749 ( Q14749 ) catabolizes S-adenosylmethionine ( DB00118 ) , the main methyl donor of the body . Patients with cirrhosis show attenuated Q14749 expression , which is absent in hepatocellular carcinoma ( HCC ) samples . Q14749 (-/-) mice develop spontaneous steatosis that progresses to steatohepatitis , cirrhosis , and HCC . The liver is highly enriched with innate immune cells and plays a key role in the body 's host defense and in the regulation of inflammation . Chronic inflammation is the major hallmark of nonalcoholic steatohepatitis ( NASH ) progression . The aim of our study was to uncover the molecular mechanisms leading to liver chronic inflammation in the absence of Q14749 , focusing on the implication of natural killer ( NK ) / natural killer T ( NKT ) cells . We found increased expression of T helper (Th)1- over Th2-related cytokines , tumor necrosis factor ( P01375 ) -related apoptosis-inducing ligand ( P50591 ) -R2/DR5 , and several ligands of NK cells in Q14749 (-/-) livers . Interestingly , NK cells from Q14749 (-/-) mice were spontaneously activated , expressed more P50591 , and had strong cytotoxic activity , suggesting their contribution to the proinflammatory environment in the liver . Accordingly , NK cells mediated hypersensitivity to concanavalin A ( ConA ) -mediated hepatitis in Q14749 (-/-) mice . Moreover , Q14749 (-/-) mice were hypersensitive to endotoxin-mediated liver injury . NK cell depletion and adoptive transfer of P50591 (-/-) liver-NK cells protected the liver against lipopolysaccharide ( LPS ) liver damage . CONCLUSION : Our data allow us to conclude that P50591 -producing NK cells actively contribute to promote a proinflammatory environment at early stages of fatty liver disease , suggesting that this cell compartment may contribute to the progression of NASH . 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 . Structure and Expression Analyses of DB04031 Elements in Relation to Functional Genes . SINE-VNTR-Alu ( DB04031 ) elements are present in hominoid primates and are divided into 6 subfamilies ( DB04031 -A to DB04031 -F ) and active in the human population . Using a bioinformatic tool , 22 DB04031 element-associated genes are identified in the human genome . In an analysis of genomic structure , DB04031 elements are detected in the 5' untranslated region ( UTR ) of Q68CP4 ( DB04031 -B ) , Q96LB0 ( DB04031 -D ) , Q12794 ( DB04031 -F ) , Q07283 ( DB04031 -F ) , and Q8WWM7 ( DB04031 -F ) genes , while some elements are observed in the 3'UTR of Q8N0Z3 ( DB04031 -B ) , Q9Y2W6 ( DB04031 -C ) , O95249 ( DB04031 -D ) , Q8N3I7 ( DB04031 -D ) , Q6P3R8 ( DB04031 -D ) , P08910 ( DB04031 -F ) , Q9BXJ2 ( DB04031 -F ) , Q9Y5N6 ( DB04031 -F ) , Q5SWH9 ( DB04031 -F ) , and Q6PK04 ( DB04031 -F ) genes . They could contribute to exon extension or supplying poly A signals . LEPR ( DB04031 -C ) , P09917 ( DB04031 -D ) , Q9NTI5 ( DB04031 -D ) , and Q8WWZ4 ( DB04031 -F ) genes also showed alternative transcripts by DB04031 exonization events . Dominant expression of HYAL1_SVA appeared in lung tissues , while HYAL1_noSVA showed ubiquitous expression in various human tissues . Expression of both transcripts ( TDRKH_SVA and TDRKH_noSVA ) of the Q9Y2W6 gene appeared to be ubiquitous . Taken together , these data suggest that DB04031 elements cause transcript isoforms that contribute to modulation of gene regulation in various human tissues . 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 . Expression of liver X receptor alpha and lipid metabolism in granulocyte-macrophage colony-stimulating factor-induced human monocyte-derived macrophage . Liver X receptor ( LXR ) is a nuclear receptor that acts as a sterol sensor and metabolic regulator of cholesterol and lipid homeostasis . The foam cell transformation of macrophages ( Mvarphi ) is considered a critical process in atherosclerotic lesions . The relationship , however , of the foam cell transformation of Mvarphi and LXR is not fully understood . The purpose of the present study was to examine the expression of LXRalpha , retinoid X receptor (RXR)alpha , DB00171 -binding cassette transporter ( O95477 ) , and macrophage scavenger receptor A ( Q9UBK8 -A ) , and lipid accumulation in human monocyte-derived Mvarphi . The expression of LXRalpha , O95477 , Q9UBK8 -A in 7 day cultured granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) -induced Mvarphi ( GM-Mvarphi ) was significantly higher than that in 7 day cultured P09603 -induced Mvarphi ( M-Mvarphi ) . The expression levels of LXRalpha , O95477 and Q9UBK8 -A protein decreased from 48 h to 5 days after the addition of lipopolysaccharide ( LPS ) in GM-Mvarphi , but only Q9UBK8 -A protein decreased at 5 days after the addition of LPS in M-Mvarphi . Intracellular lipid accumulation was clearly observed when GM-Mvarphi was pre-stimulated with LPS for 48 h and incubated with oxidized LDL for an additional 5 days . These findings suggest that the inhibitory activity of LXRalpha , O95477 and Q9UBK8 -A by LPS may be related to the transformation of Mvarphis , especially GM-Mvarphi into foam cells . 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 . P22309 *28 is associated with greater decrease in serum K⁺ levels following oral intake of procaterol . BACKGROUND AND OBJECTIVE : DB01366 is a potent β2-agonist frequently used for the management of asthma and chronic obstructive pulmonary disease . The efficacy and adverse effects of β2-agonists are heterogeneous in individual patients , which may be partly caused by genetic variations in metabolizing enzymes and receptor molecules . The present study was designed to analyze the relationship between gene polymorphisms and physiological effects of procaterol in healthy subjects . METHODS : Ninety-two non-smoking healthy volunteers were given 1 µg/kg body weight ( max 50 µg ) of procaterol as a dry syrup preparation , and the serum concentrations of procaterol , serum K(+) , and the physical responses were monitored for 240 min . We genotyped β2-adrenergic receptor ( P07550 ) ( Arg16Gly and Gln27Glu ) , cytochrome P450 3A4 ( rs2246709 , rs4646437 ) , and uridine diphosphate glucuronosyltransferase 1A1 ( P22309 ) ( rs4148323 [ allele A , *6 ] , rs12479045 , rs4148328 , rs4663971 , rs12052787 , rs4148329 , A (TA)6/7 TAA [ seven-repeat allele , *28 ] ) . DB01366 concentrations in serum were measured by liquid chromatography-tandem mass spectrometry . RESULTS : No gene polymorphisms affected serum procaterol concentrations . Meanwhile , overall serum K(+) level changes were significantly lower in carriers of P22309 *28 than in non-carriers after correcting for strong effects of serum procaterol concentrations and baseline K(+) levels . No other polymorphisms were associated with serum K(+) levels . None of polymorphisms of P07550 were associated with any physical responses . CONCLUSION : The present study indicates that significant hypokalemia may occur in carriers of P22309 *28 by systemic administration of procaterol and potentially by other β2-agonists metabolized in the liver . 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 . 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 . DB00134 sulfoxide reductase A and a dietary supplement S-methyl-L-cysteine prevent Parkinson's-like symptoms . Parkinson 's disease ( PD ) , a common neurodegenerative disease , is caused by loss of dopaminergic neurons in the substantia nigra . Although the underlying cause of the neuronal loss is unknown , oxidative stress is thought to play a major role in the pathogenesis of PD . The amino acid methionine is readily oxidized to methionine sulfoxide , and its reduction is catalyzed by a family of enzymes called methionine sulfoxide reductases ( MSRs ) . The reversible oxidation-reduction cycle of methionine involving MSRs has been postulated to act as a catalytic antioxidant system protecting cells from oxidative damage . Here , we show that one member of the Q9UBK8 family , Q9UJ68 , inhibits development of the locomotor and circadian rhythm defects caused by ectopic expression of human alpha-synuclein in the Drosophila nervous system . Furthermore , we demonstrate that one way to enhance the Q9UJ68 antioxidant system is dietary supplementation with S-methyl-L-cysteine ( SMLC ) , found abundantly in garlic , cabbage , and turnips . SMLC , a substrate in the catalytic antioxidant system mediated by Q9UJ68 , prevents the alpha-synuclein-induced abnormalities . Therefore , interventions focusing on the enzymatic reduction of oxidized methionine catalyzed by Q9UJ68 represent a new prevention and therapeutic approach for PD and potentially for other neurodegenerative diseases involving oxidative stress . 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 . 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 . Embryo transfer induces a subclinical endometritis in recipient mares which can be prevented by treatment with non-steroid anti-inflammatory drugs . We tested the hypothesis that subclinical endometritis occurs after embryo transfer ( ET ) in the horse . Recipient mares were treated with meclofenamic acid ( M ) or flunixin meglumin ( F ) after ET or were left untreated ( n=9 per group ) . Embryos were re-collected 4 days after transfer . Endometrial biopsies were taken for histology and analysis of cyclooxygenase-2 ( P35354 ) by immunohistochemistry and for PCR . Bacteriological swabs were collected from the uterus and lavage fluid of donor and recipient mares . Progesterone and prostaglandin F(2alpha) release was analysed in recipient mares after ET . Four days after ET , four embryos were recovered from group M and three from group F and untreated mares , each . The number of polymorph nuclear neutrophils was reduced in treated mares ( p < 0.05 ) . Expression of mRNA for inflammatory cytokines did not differ between groups . In group M , expression of endometrial prostaglandin-E-synthase was higher than in group F ( p < 0.05 ) . Three out of nine control mares underwent preterm luteolysis ( p < 0.05 vs. treatment groups ) , prostaglandin release ( p < 0.05 ) and the number of P35354 positive cells ( p < 0.01 ) were significantly higher than in treated mares . Only few bacteriological swabs were positive . In conclusion , treatment of embryo recipient mares with non-steroid anti-inflammatory drugs inhibits the inflammatory response of the endometrium after ET . DB00939 may have advantages in comparison to flunixin meglumin due to a different influence on prostaglandin synthesis that may not result in inhibition of embryonic mobility . Q03135 tyrosine phosphorylation enhances paclitaxel-mediated cytotoxicity . Q03135 ( Q03135 ) , a highly conserved membrane-associated protein , is a putative regulator of cellular transformation . Q03135 is localized in the plasmalemma , secretory vesicles , Golgi , mitochondria , and endoplasmic reticulum membrane and associates with the microtubule cytoskeleton . Taxanes such as paclitaxel ( DB01229 ) are potent anti-tumor agents that repress the dynamic instability of microtubules and arrest cells in the G(2)/M phase . Src phosphorylation of DB00135 -14 on Q03135 regulates its cellular localization and function . We report that phosphorylation of Q03135 on DB00135 -14 regulates paclitaxel-mediated apoptosis in MCF-7 breast cancer cells . Befitting its role as a multitasking molecule , we show that Q03135 sensitizes cells to apoptosis by regulating cell cycle progression and activation of the apoptotic signaling molecules P10415 , p53 , and P38936 . We demonstrate that phosphorylated Q03135 triggers apoptosis by inactivating P10415 and increasing mitochondrial permeability more efficiently than non-phosphorylated Q03135 . Furthermore , expression of P38936 , which correlates with taxane sensitivity , is regulated by Q03135 phosphorylation in a p53-dependent manner . Collectively , our findings underscore the importance of Q03135 phosphorylation in apoptosis and suggest that events that negate Q03135 tyrosine phosphorylation may contribute to anti-microtubule drug resistance . 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 . [ Acute transverse myelitis after obstetric epidural anesthesia ] . INTRODUCTION : Acute transverse myelitis is an acute or subacute disorder of the spinal cord resulting in motor , sensory and sphincter dysfunction secondary to various causes . CASE REPORT : We present a 32 year-old female patient with an acute episode of bladder dysfunction and fever , followed by motor and sensory dysfunction in legs with sensory spinal level at D2-D3 , two weeks after an eutocic delivery with uncomplicated epidural anesthesia . The cerebrospinal fluid ( P04141 ) showed mild lymphocytic pleocytosis , high protein levels with normal glucose concentration , absence of oligoclonal bands and negative serum and P04141 virology screening . The cervicodorsal magnetic resonance imaging showed widening of the spinal cord with diffuse patchy hyperintensity on the P13671 -D1 and D2-D5 levels without contrast enhancement . The patient was treated with intravenous high doses of methylprednisolone with favorable outcome and complete recovery within one year and no relapses two years after the episode . DISCUSSION : The main etiologies of non-compressive acute myelopathy as multiple sclerosis , systemic diseases , spinal cord infarct and direct infections have been ruled out with the complementary examinations . We consider that our patient had a parainfectious acute transverse myelitis and epidural anesthesia could be an incidental but possible contributing factor . 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 ROS-activated p38MAPK pathway is involved in the protective effect of H2S against chemical hypoxia-induced inflammation in PC12 cells . We have demonstrated the neuroprotection of hydrogen sulfide ( H2S ) against chemical hypoxia-induced injury by inhibiting p38MAPK pathway . The present study attempts to evaluate the effect of H2S on chemical hypoxia-induced inflammation responses and its mechanisms in PC12 cells . We found that treatment of PC12 cells with cobalt chloride ( CoCl2 , a hypoxia mimetic agent ) enhanced P05231 secretion , nitric oxide ( NO ) generation and expression levels of inducible nitric oxide synthase ( P35228 ) and neuronal nitric oxide synthase ( P29475 ) . L-canavanine , a selective P35228 inhibitor , partly blocked CoCl2-induced cytotoxicity , apoptosis and mitochondrial insult . In addition , DB02207 ( 7-NI ) , an inhibitor of P29475 , also partly attenuated the CoCl2-induced cytotoxicity . The inhibition of p38MAPK by SB203580 ( a selective p38MAPK inhibitor ) or genetic silencing of p38MAPK by RNAi ( Si-p38 ) depressed not only CoCl2-induced P35228 expression , NO production , but also P05231 secretion . In addition , N-acetyl-L-cysteine , a reactive oxygen species ( ROS ) scavenger , conferred a similar protective effect of SB203580 or Si-p38 against CoCl2-induced inflammatory responses . Importantly , pretreatment of PC12 cells with exogenous application of sodium hydrosulfide ( a H2S donor , 400 μmol/l ) for 30 min before exposure to CoCl2 markedly attenuated chemical hypoxia-stimulated P35228 and P29475 expression , NO generation and P05231 secretion as well as p38MAPK phosphorylation in PC12 cells . Taken together , we demonstrated that p38MAPK- P35228 pathway contributes to chemical hypoxia-induced inflammation and that H2S produces an anti-inflammatory effect in chemical hypoxia-stimulated PC12 cells , which may be partly due to inhibition of ROS-activated p38MAPK- P35228 pathway . 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 .

MH_train_1516

MH_train_1516

MH_train_1516

interacts_with DB00031?

multiple_choice

[ 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 . Imatinib blocks migration and invasion of medulloblastoma cells by concurrently inhibiting activation of platelet-derived growth factor receptor and transactivation of epidermal growth factor receptor . Platelet-derived growth factor ( PDGF ) receptor ( P09619 ) expression correlates with metastatic medulloblastoma . PDGF stimulation of medulloblastoma cells phosphorylates extracellular signal-regulated kinase ( P29323 ) and promotes migration . We sought to determine whether blocking P09619 activity effectively inhibits signaling required for medulloblastoma cell migration and invasion . DAOY and D556 human medulloblastoma cells were treated with imatinib mesylate ( Gleevec ) , a P09619 tyrosine kinase inhibitor , or transfected with small interfering RNA ( siRNA ) to P09619 to test the effects of blocking P09619 phosphorylation and expression , respectively . P09619 cell signaling , migration , invasion , survival , and proliferation following DB00102 stimulation , with and without P09619 inhibition , were measured . DB00102 treatment of cells increased P09619 , Akt and P29323 phosphorylation , and transactivated epidermal growth factor receptor ( P00533 ) , which correlated with enhanced migration , survival , and proliferation . Imatinib ( 1 μmol/L ) treatment of DAOY and D556 cells inhibited DB00102 - and serum-mediated migration and invasion at 24 and 48 h , respectively , and concomitantly inhibited DB00102 activation of P09619 , Akt , and P29323 but increased P60484 expression and activity . Imatinib treatment also induced DAOY cell apoptosis at 72 h and inhibited DAOY and D556 cell proliferation at 48 h . siRNA silencing of P09619 similarly inhibited signaling , migration , and survival and both siRNA and imatinib treatment inhibited DB00102 -mediated P00533 transactivation , indicating that the effects of imatinib treatment are specific to P09619 target inhibition . These results indicate that P09619 tyrosine kinase activity is critical for migration and invasion of medulloblastoma cells possibly by transactivating P00533 ; thus , imatinib may represent an important novel therapeutic agent for the treatment of medulloblastoma . DB06155 reduces obesity-associated hepatic steatosis and features of metabolic syndrome in obese Zucker fa/fa rats . This study investigated the effects of rimonabant ( SR141716 ) , an antagonist of the cannabinoid receptor type 1 ( P21554 ) , on obesity-associated hepatic steatosis and related features of metabolic syndrome : inflammation ( elevated plasma levels of tumor necrosis factor alpha [ TNFalpha ] ) , dyslipidemia , and reduced plasma levels of adiponectin . We report that oral treatment of obese ( fa/fa ) rats with rimonabant ( 30 mg/kg ) daily for 8 weeks abolished hepatic steatosis . This treatment reduced hepatomegaly , reduced elevation of plasma levels of enzyme markers of hepatic damage ( alanine aminotransferase , gamma glutamyltransferase , and alkaline phosphatase ) and decreased the high level of local hepatic TNFalpha currently associated with steatohepatitis . In parallel , treatment of obese ( fa/fa ) rats with rimonabant reduced the high plasma level of the proinflammatory cytokine TNFalpha and increased the reduced plasma level of the anti-inflammatory hormone adiponectin . Finally , rimonabant treatment also improved dyslipidemia by both decreasing plasma levels of triglycerides , free fatty acids , and total cholesterol and increasing the HDLc/LDLc ratio . All the effects of rimonabant found in this study were not or only slightly observed in pair-fed obese animals , highlighting the additional beneficial effects of treatment with rimonabant compared to diet . These results demonstrate that rimonabant plays a hepatoprotective role and suggest that this P21554 receptor antagonist potentially has clinical applications in the treatment of obesity-associated liver diseases and related features of metabolic syndrome . 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 . Candidate genetic markers and the risk of restenosis after coronary angioplasty . The aim of the present study was to test for possible associations between candidate gene polymorphisms and the risk of restenosis and recurrent restenosis after percutaneous transluminal coronary angioplasty ( PTCA ) without stenting . We followed up 511 PTCA patients , and restenosis and recurrent restenosis were defined according to angiographical criteria . Genotyping of the beta-fibrinogen -455 G/A , glycoprotein ( GP ) IIIa PlA1/PlA2 , plasminogen activator inhibitor-1 ( P05121 ) 4G/5G , factor V Leiden 1691 G/A , tumour necrosis factor alpha ( TNFalpha ) -238 G/A , TNFalpha -308 G/A , interleukin ( IL ) -1alpha -889 C/T , IL-1beta -511 C/T , methylenetetrahydrofolate reductase ( P42898 ) 677 C/T and endothelial nitric oxide synthase ( P29474 ) 4 b/a gene polymorphisms was performed by PCR and restriction-fragment-length-polymorphism-based techniques . One hundred and sixty patients ( 31.3 % ) developed restenosis and in 130 of these patients , of whom 123 were available for analysis , a second PTCA without stenting was performed . Of these patients , 35 ( 28.5 % ) developed recurrent restenosis . None of the investigated genotypes were associated with the risk of restenosis or recurrent restenosis after PTCA . The degree of stenosis before and immediately after PTCA and the severity of the lesion were independent predictors for restenosis after PTCA . In conclusion , there was no association between the beta-fibrinogen -455 G/A , GP IIIa PlA1/A2 , P05121 4G/5G , factor V Leiden 1691 G/A , TNFalpha -238 G/A , TNFalpha -308 G/A , IL-1alpha -889 C/T , the IL-1beta -511 C/T , P42898 677 C/T and P29474 4 b/a gene polymorphisms and the risk of restenosis after PTCA as well as recurrent restenosis after repeated PTCA . 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 . Specific expression of matrix metalloproteinases 1 , 3 , 9 and 13 associated with invasiveness of breast cancer cells in vitro . Several matrix metalloproteinases ( MMPs ) and tissue inhibitors of MMPs ( TIMPs ) were studied in highly invasive ( MDA-MB-231 ) and slightly invasive ( MCF-7 , T47D , BT-20 ) breast cancer cell lines . Investigations were carried out at the protein level and/or at the mRNA level , either in cells cultured as monolayers on plastic , or in cells seeded on a thin layer of Matrigel basement membrane matrix . Analysis of MMP expression by RT-PCR showed expression of P03956 . P08254 , and P45452 in highly invasive MDA-MB-231 cells , but not in slightly invasive cell lines . The extracellular secretion of P03956 and P08254 by MDA-MB 231 cells could be also shown by ELISA . P01033 and P16035 mRNAs were found in all cell lines , however , the extracellular secretion of both TIMPs was much higher in MDA-MB-231 cells than in the other cell lines . When the cells were cultured on Matrigel matrix , P14780 expression was induced in MDA-MB-231 cells only , as assessed by RT-PCR and zymography experiments . The invasive potential of MDA-MB-231 cells evaluated in vitro through Matrigel was significantly inhibited by the MMP inhibitor DB00786 , by 25 % and 50 % at the concentrations of 2 x 10(-6) M and 10(-5) M , respectively . In conclusion , our data show that highly invasive MDA-MB-231 cells but not slightly invasive T47D , MCF-7 and BT-20 cells express P03956 , P08254 , P14780 and P45452 . P14780 which is specifically up-regulated by cell contact to Matrigel , may play a key role in the invasiveness of MDA-MB-231 cells through basement membranes . 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 . P01258 gene-related peptide stimulates proliferation of human endothelial cells . The effects of the vasoactive perivascular neuropeptides calcitonin gene-related peptide ( P80511 ) , neurokinin A ( P20366 ) , neuropeptide Y ( P01303 ) , and vasoactive intestinal polypeptide ( P01282 ) on proliferation of cultured human umbilical vein endothelial cells ( HUVECs ) were investigated . P80511 was shown to increase both cell number and DNA synthesis , whereas P20366 , P01303 , and P01282 were ineffective . 125I-labeled P80511 was shown to bind to HUVECs and this binding was displaced by addition of unlabeled P80511 , suggesting the existence of specific P80511 receptors . The effect of P80511 on formation of adenosine 3',5'-cyclic monophosphate ( DB02527 ) and inositol phosphates ( InsP ) , two intracellular messengers known to be involved in regulation of cell proliferation , was investigated . P80511 stimulated DB02527 formation but was without effect on the formation of InsP . Proliferation , as well as DB02527 formation , was also stimulated by cholera toxin . P09038 stimulated growth without affecting DB02527 or InsP formation , whereas thrombin , which increased InsP formation , did not stimulate proliferation . We thus suggest that P80511 may act as a local factor stimulating proliferation of endothelial cells ; that the mechanism of action is associated with DB02527 formation ; and that this effect of P80511 may be important for formation of new vessels during physiological and pathophysiological events such as ischemia , inflammation , and wound healing . Preoperative radiotherapy and extracellular matrix remodeling in rectal mucosa and tumour matrix metalloproteinases and plasminogen components . BACKGROUND. Preoperative radiotherapy reduces recurrence but increases postoperative morbidity . The aim of this study was to explore the effect of radiotherapy in rectal mucosa and rectal tumour extracellular matrix ( Q13201 ) by studying enzymes and growth factors involved in Q13201 remodeling . MATERIALS AND METHODS . Twenty patients with short-term preoperative radiotherapy and 12 control patients without radiotherapy were studied . Biopsies from rectal mucosa and tumour were collected prior to radiotherapy and at surgery . Tissue P03956 , -2 , -9 , P01033 , uPA , P05121 , TGF-beta1 and calprotectin were determined by ELISA . Biopsies from irradiated and non-irradiated peritoneal areas were also analysed . RESULTS . Radiotherapy increased the tissue levels of P08253 and P05121 in both the rectal mucosa and tumours while calprotectin and uPA showed an increase only in the mucosa after irradiation . The increase of calprotectin was due to an influx of inflammatory cells as revealed by immunohistochemistry . Prior to irradiation , the tumour tissues had increased levels of P03956 , -2 , -9 , total TGF-beta1 , uPA , P05121 and calprotectin compared to mucosa , while P01033 and the active TGF-beta1 fraction showed no statistical difference . CONCLUSIONS . This study indicates a radiation-induced effect on selected Q13201 remodeling proteases . This reaction may be responsible for early and late morbidity . Interference of this response might reduce these consequences . Leishmania major protein disulfide isomerase as a drug target : enzymatic and functional characterization . Leishmaniasis is a major health problem worldwide and tools available for their control are limited . Effective vaccines are still lacking , drugs are toxic and expensive , and parasites develop resistance to chemotherapy . In this context , new antimicrobials are urgently needed to control the disease in both human and animal . Here , we report the enzymatic and functional characterization of a Leishmania virulence factor , Leishmania major Protein disulfide isomerase ( LmPDI ) that could constitute a potential drug target . LmPDI possesses domain structure organization similar to other P07237 family members ( a , a ' , b , b ' and c domains ) , and it displays the three enzymatic and functional activities specific of P07237 family members : isomerase , reductase and chaperone . These results suggest that LmPDI plays a key role in assisting Leishmania protein folding via its capacity to catalyze formation , breakage , and rearrangement of disulfide bonds in nascent polypeptides . Moreover , DB00626 , a reductase activity inhibitor , and DB03615 , a chaperone activity inhibitor , were tested in LmPDI enzymatic assays and versus Leishmania promastigote in vitro cultures and Leishmania amastigote multiplication inside infected THP-1-derived macrophages . DB00626 inhibited both isomerase and reductase activities , while DB03615 had no effect on the chaperone activity . Interestingly , DB00626 blocked in vitro promastigote growth as well as amastigote multiplication inside macrophages with EC(50) values of 39 μM . These results suggest that LmPDI may constitute an interesting target for the development of new anti-Leishmania drugs . 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 . Echinococcus granulosus : biological comparison of cattle isolates from endemic regions of Argentina and Spain . In the present study we have compared cattle isolates of Echinococcus granulosus from Argentina and Spain . The aim was to compare and determine if there exist phenotypic and genetic differences within E. granulosus cattle isolates between an endemic area of Spain ( where the disease is mainly restricted to a sheep-dog cycle ) and an endemic area of Argentina ( where cattle are the most abundant intermediate hosts ) . The Spanish samples were previously identified as P55008 genotype . The Argentinean samples were also identified as P55008 , but some variants were found for the cytochrome c oxidase-1 ( CO1 ) and DB00157 dehydrogenase-1 ( P03886 ) mitochondrial genes . When comparing the cyst features and the morphology of the larval rostellar hooks in both regions , some differences were found . The morphometric analyses of the larval rostellar hooks showed the existence of two distinct clearly separated groups ( one corresponding to the Argentinean samples and the other to the Spanish ones ) . In conclusion , there are some genetic and phenotypic differences within E. granulosus cattle isolates from Argentina and Spain . Probably these differences , more important from an epidemiological point of view , are related to different steps in the disease control in both countries . Further studies involving other epidemiological , morphometric and molecular data , including other types of livestock , would contribute to clarify and expand the present work . The FGF receptor uses the endocannabinoid signaling system to couple to an axonal growth response . A key role for DAG lipase activity in the control of axonal growth and guidance in vitro and in vivo has been established . For example , DAG lipase activity is required for FGF-stimulated calcium influx into neuronal growth cones , and this response is both necessary and sufficient for an axonal growth response . The mechanism that couples the hydrolysis of DAG to the calcium response is not known . The initial hydrolysis of DAG at the sn-1 position ( by DAG lipase ) will generate 2-arachidonylglycerol , and this molecule is well established as an endogenous cannabinoid receptor agonist in the brain . In the present paper , we show that in rat cerebellar granule neurons , P21554 cannabinoid receptor antagonists inhibit axonal growth responses stimulated by P19022 and P09038 . Furthermore , three P21554 receptor agonists mimic the P19022 / P09038 response at a step downstream from FGF receptor activation , but upstream from calcium influx into cells . In contrast , we could find no evidence for the P21554 receptor coupling the TrkB neurotrophin receptor to an axonal growth response in the same neurons . The observation that the P21554 receptor can couple the activated FGF receptor to an axonal growth response raises novel therapeutic opportunities . Rotenone remarkably attenuates oxidative stress , inflammation , and fibrosis in chronic obstructive uropathy . Mitochondrial abnormality has been shown in many kidney disease models . However , its role in the pathogenesis of chronic kidney diseases ( CKDs ) is still uncertain . In present study , a mitochondrial complex I inhibitor rotenone was applied to the mice subjected to unilateral ureteral obstruction ( UUO ) . Following 7-days rotenone treatment , a remarkable attenuation of tubular injury was detected by DB00233 staining . In line with the improvement of kidney morphology , rotenone remarkably blunted fibrotic response as shown by downregulation of fibronectin ( FN ) , plasminogen activator inhibitor-1 ( P05121 ) , collagen I , collagen III , and α-SMA , paralleled with a substantial decrease of TGF-β 1 . Meanwhile , the oxidative stress markers thiobarbituric acid-reactive substances ( TBARS ) and heme oxygenase 1 ( P09601 ) and inflammatory markers P01375 -α , IL-1β , and P05362 were markedly decreased . More importantly , the reduction of mitochondrial DNA copy number and mitochondrial P03886 ( mtND1 ) expression in obstructed kidneys was moderately but significantly restored by rotenone , suggesting an amelioration of mitochondrial injury . Collectively , mitochondrial complex I inhibitor rotenone protected kidneys against obstructive injury possibly via inhibition of mitochondrial oxidative stress , inflammation , and fibrosis , suggesting an important role of mitochondrial dysfunction in the pathogenesis of obstructive kidney disease . 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 . Modulation of cyclin D1 expression in human tumoral parathyroid cells : effects of growth factors and calcium sensing receptor activation . The study investigated cyclin D1 regulation by growth factors and calcium sensing receptor ( P41180 ) in human tumoral parathyroid cells . Basic fibroblast and epidermal growth factors increased cyclin D1 and phosphorylated extracellular signal-regulated kinases ( pERK1/2 ) levels that were both efficiently inhibited by P41180 agonists . By contrast , in growth factors-free medium cyclin D1 levels were either unaffected or stimulated by P41180 activation independently from P27361 /2 pathway . Transforming growth factor beta ( TGFbeta ) reduced cyclin D1 levels in the majority of tumors , this effect being not influenced by P41180 activation and menin expression levels . In conclusion , in parathyroid tumors cyclin D1 expression was modulated by growth factors and P41180 activation . These data further support the oncogenic role of cyclin D1 , which resulted to be target for stimulation by P09038 and P01133 and inhibition by P41180 and TGFbeta signalling in the parathyroid . 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 . 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 . Disulfide isoform intermediates in the reoxidation of recombinant human basic fibroblast growth factor . The reoxidation of human recombinant basic fibroblast growth factor was investigated following treatment of the protein with a mixture of reduced and oxidized glutathione , both in the absence and in the presence of protein disulfide isomerase . The oxidative process took place throughout the formation of two transient intermediates and yielded a stable P09038 derivative , P41247 - P09038 . All of these components were separated by HPLC and accurately characterized at the molecular level by advanced mass spectrometric procedures . When the reoxidation was carried out in the presence of P07237 , a 4-fold increase in the reaction rate was estimated . A mixed disulfide with a single glutathione molecule was shown to occur in the two transient intermediates , each of which has different cysteine residues involved in the linkage . The final product P41247 - P09038 was structurally different from other P09038 derivatives previously described [ Thompson , S. A. ( 1992 ) J. Biol. Chem. 267 , 2269-2273 ; Caccia et al. ( 1992 ) Eur. J. Biochem. 204 , 649-655 ] . The four cysteine residues are all involved in disulfide bridges ; DB00151 34 and DB00151 78 are linked to exogenous glutathione , whereas DB00151 91 and DB00151 101 form an intramolecular S-S bridge .

MH_train_1517

MH_train_1517

MH_train_1517

interacts_with DB08907?

multiple_choice

Retinoic acid receptors in retinoid responsive ovarian cancer cell lines detected by polymerase chain reaction following reverse transcription . The growth inhibitory effects of all-trans and 13-cis retinoic acid ( RA ) and of the synthetic retinoids DB02877 , DB02877 -ethylester and TTNN were studied on seven human epithelial ovarian cancer cell lines and one ovarian teratocarcinoma cell line . Six of seven ovarian adenocarcinoma cell lines were inhibited in their growth by RA and by synthetic retinoids in a dose dependent manner . No response to these substances was observed for the ovarian teratocarcinoma cell line . The knowledge that RA and retinoids exert their action on the cells via nuclear receptors led us to examine the expression of P10276 , -beta and -gamma mRNA by these cell lines by polymerase chain reaction following reverse transcription . All cell lines expressed P10276 and -gamma mRNA and six of the eight cell lines were found to express additionally P10826 mRNA , among them the ovarian teratocarcinoma cell line . Our data indicate that there was no direct association between the presence of RAR subtype transcripts and the response to retinoids in ovarian cancer cell lines . Structural elucidation of DB00007 and its analogues in solution : insight into their bioactive conformation . DB00007 [ DLeu6 , NHEt10 ] DB00644 , a potent gonadotropin-releasing hormone ( DB00644 ) agonist , is used in a wide variety of hormone-related diseases like cancer and endometriosis . In this report , the conformational behaviour of DB00007 and its linear synthetic analogues , namely [ Tyr5(OMe) , DLeu6 , Aze9 , NHEt10 ] DB00644 ( 1 ) and [ Tyr5(OMe) , DLeu6 , NHEt10 ] DB00644 ( 2 ) have been studied in DB01093 and H2O solutions by means of 2D nuclear magnetic resonance ( NMR ) experiments and detailed molecular dynamics ( MD ) simulations . The aim was to identify the conformational requirements of DB00644 analogues for agonistic activity . This approach is of value as no crystallographic data are available for the P30968 ( G protein-coupled receptor , GPCR ) . The NOE data indicate the existence of a β-turn type I in the 2-5 segments of DB00007 and its linear analogues in the case of using DB01093 -d6 as solvent , whereas a β-turn type II in the 3-6 segments is indicated using D2O as solvent . The final structures fulfil the conformational requirements that are known , in the literature , to play a significant role in receptor recognition and activation . Finally , the linear analogues ( 1 ) and ( 2 ) are biologically active when tested against the human breast cancer cell line , MCF-7 . An analysis of the effects of retinoic acid and other retinoids on the development of adrenergic cells from the avian neural crest . In the present work , we have investigated the role of all-trans-retinoic acid ( all-trans RA ) , and several other natural and synthetic retinoids , in the development of adrenergic cells in quail neural crest cultures . Dose response studies using all-trans RA and 13-cis RA revealed a dose-dependent increase in the number of adrenergic cells in neural crest cultures . Similar dose response studies using RA isomers and other natural retinoids did not result in the same increases . In order to determine the receptor mediating the effects of all-trans RA in the neural crest , we tested several synthetic analogs which specifically bind to a particular RA receptor ( RAR ) subtype . We found that the compound AM 580 , which activates the P10276 , produced an increase in adrenergic cells similar to that seen with all-trans RA . The compound DB02877 , which activates all RAR subtypes , also resulted in an increase in adrenergic cells . We conclude that the increase in adrenergic cells seen with all-trans RA is mediated by P10276 and possibly P10826 . To further define the actions of all-trans RA on the neural crest we incubated cultures with 5-bromo-2'-deoxyuridine ( BrdU ) to determine whether all-trans RA could affect the rate of proliferation . The results show that while all-trans RA did not increase the fraction of cells incorporating BrdU into their nuclei at early time points ( 24 h ) , it did increase BrdU incorporation by tyrosine hydroxylase ( TH ) positive cells at 5 days in culture . These findings demonstrate that the increase in adrenergic cells seen with all-trans RA in neural crest cultures is likely due to an increase in the proliferation of cells already expressing TH . Time-Qualified Patterns of Variation of PPARγ , P26358 , and Q9UBC3 Expression in Pancreatic Cancer Cell Lines . Carcinogenesis is related to the loss of homeostatic control of cellular processes regulated by transcriptional circuits and epigenetic mechanisms . Among these , the activities of peroxisome proliferator-activated receptors ( PPARs ) and DNA methyltransferases ( DNMTs ) are crucial and intertwined . PPARγ is a key regulator of cell fate , linking nutrient sensing to transcription processes , and its expression oscillates with circadian rhythmicity . Aim of our study was to assess the periodicity of PPARγ and DNMTs in pancreatic cancer ( PC ) . We investigated the time-related patterns of P37231 , P26358 , and Q9UBC3 expression monitoring their mRNA levels by qRT-PCR at different time points over a 28-hour span in BxPC-3 , CFPAC-1 , PANC-1 , and MIAPaCa-2 PC cells after synchronization with serum shock . P37231 and P26358 expression in PANC-1 cells and P37231 expression in MIAPaCa-2 cells were characterized by a 24 h period oscillation , and a borderline significant rhythm was observed for the P37231 , P26358 , and Q9UBC3 expression profiles in the other cell lines . The time-qualified profiles of gene expression showed different shapes and phase relationships in the PC cell lines examined . In conclusion , P37231 and DNMTs expression is characterized by different time-qualified patterns in cell lines derived from human PC , and this heterogeneity could influence cell phenotype and human disease behaviour . DB11320 -induced P05231 and P10145 production are differentially modulated by P01579 and P05112 in human keratinocytes . It is known that large amounts of histamine are stored in mast cells located in the superficial dermis of the skin and can be released upon appropriate stimulation . However , the effects of histamine on keratinocyte function have not been well characterized . We therefore examined the capacity of histamine to modulate the production of interleukin ( IL ) -6 and P10145 by keratinocytes . We found that histamine significantly augmented the production of P05231 and P10145 in a dose- and time-dependent manner . The enhancing effects of histamine were completely inhibited by a potent H1 receptor ( P35367 ) antagonist , emedastine difumarate . Pyrilamine ( a much weaker P35367 antagonist ) and cimetidine ( an P25021 antagonist ) only partially inhibited the enhancing effects of histamine . The histamine-induced up-regulation of P05231 and P10145 production , however , was completely abrogated by a combination of DB06691 and cimetidine . The P05231 production was significantly enhanced by interferon ( IFN ) -gamma . Interestingly , P01579 and P05112 both significantly augmented the histamine-induced P05231 production . On the other hand , the production of P10145 was inhibited by P01579 , and P01579 and P05112 both completely abrogated the histamine-induced P10145 production . These results suggest that the histamine-induced P05231 production and P10145 production are differentially regulated by P01579 and P05112 . DB11320 may be an important modulator of cytokine production in epidermal milieu . 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 . Specific properties of a C-terminal truncated androgen receptor detected in hormone refractory prostate cancer . Mutations in the human androgen receptor ( AR ) gene that lead to C-terminus truncated AR variants are frequently detected in prostate cancer ( PC ) . These AR variants lack both the ligand-binding domain ( LBD ) and the AF-2 region . The aim of this study was to delineate the alternative mechanisms that lead to the activation of such AR variants as they are unresponsive to hormone stimulation , and to outline consequences of the loss of the LBD/AF-2 region on their functional properties . By using an MMTV-luciferase reporter construct and LY294002 , UO126 , or ZD1839 , inhibitor of PI3K , Q02750 /2 , and P00533 signaling pathway respectively , we demonstrated that phosphorylation was required for full transcriptional activities of one these AR variants , the Q640X mutant AR . Western-blot analyses confirmed that these inhibitors affect the phosphorylation status of this AR variant . Furthermore , studies of the intranuclear colocalization of the Q640X AR with cofactors , such as CBP , Q9Y3R0 , and c-Jun , reveal that the transcriptional complex that forms around the mutant AR is different to that formed around the wild type AR . We demonstrated that CBP and c-Jun are highly recruited by the mutant AR , and this leads to an unexpected activation of AP-1 , NFAT , and NFkappaB transcriptional activities . Similar enhanced activities of these transcription factors were not observed with the wild type AR . The importance of the LBD/AF-2 for the regulation of AR transcriptional activities , the impact of the presence of such AR variants on PC cells proliferation and survival , and on progression to androgen independence are discussed . DB00928 and decitabine induce gene-specific and non-random DNA demethylation in human cancer cell lines . The DNA methyltransferase inhibitors azacytidine and decitabine represent archetypal drugs for epigenetic cancer therapy . To characterize the demethylating activity of azacytidine and decitabine we treated colon cancer and leukemic cells with both drugs and used array-based DNA methylation analysis of more than 14,000 gene promoters . Additionally , drug-induced demethylation was compared to methylation patterns of isogenic colon cancer cells lacking both DNA methyltransferase 1 ( P26358 ) and Q9UBC3 . We show that drug-induced demethylation patterns are highly specific , non-random and reproducible , indicating targeted remethylation of specific loci after replication . Correspondingly , we found that CG dinucleotides within CG islands became preferentially remethylated , indicating a role for DNA sequence context . We also identified a subset of genes that were never demethylated by drug treatment , either in colon cancer or in leukemic cell lines . These demethylation-resistant genes were enriched for Polycomb Repressive Complex 2 components in embryonic stem cells and for transcription factor binding motifs not present in demethylated genes . Our results provide detailed insights into the DNA methylation patterns induced by azacytidine and decitabine and suggest the involvement of complex regulatory mechanisms in drug-induced DNA demethylation . 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) . Involvement of Q07869 receptors in the anticonvulsant effects of a cannabinoid agonist , Q08050 55,212-2 . Cannabinoid and Q07869 receptors show well established interactions in a set of physiological effects . Regarding the seizure-modulating properties of both classes of receptors , the present study aimed to evaluate the roles of the P37231 , Q07869 and P21554 receptors on the anticonvulsant effects of Q08050 55,212-2 ( Q08050 , a non selective cannabinoid agonist ) . The clonic seizure thresholds after intravenous administration of pentylenetetrazole ( PTZ ) were assessed in mice weighing 23-30 g . Q08050 increased the seizure threshold dose dependently . Pretreatment with pioglitazone , as a PPARγ agonist , potentiated the anticonvulsant effects of Q08050 , while PPARγ antagonist inhibited these anticonvulsant effects partially . On the other hand PPARα antagonist reduced the anticonvulsant effects of Q08050 significantly . Finally the combination of P21554 antagonist and PPARα antagonist could completely block the anticonvulsant properties of Q08050 . Taken together , these results show for the first time that a functional interaction exists between cannabinoid and Q07869 receptors in the modulation of seizure susceptibility . Control of CBP co-activating activity by arginine methylation . The histone acetyltransferases CREB binding protein ( CBP ) and the related p300 protein function as key transcriptional co-activators in multiple pathways . In the case of transcriptional activation by nuclear receptors , ligand promotes the recruitment of co-activators of the P52701 family , such as Q9Y3R0 . Subsequently , the P52701 co-activators recruit other co-activators via two activation domains , P05067 and AD2 . P05067 binds CBP or p300 , whereas AD2 has been shown to activate transcription through the recruitment of the arginine methyltransferase Q86X55 . Recently , the KIX domain of CBP has been shown to be methylated by Q86X55 in vitro . Here , we report that another domain of CBP is specifically methylated by Q86X55 on conserved arginine residues in vitro and in vivo . We also provide functional evidence that arginine residues methylated by Q86X55 play a critical role in Q9Y3R0 -dependent transcriptional activation and in hormone-induced gene activation . Altogether , our data provide strong evidence that arginine methylation represents an important mechanism for modulating co-activator transcriptional activity . Oral resveratrol reduces neuronal damage in a model of multiple sclerosis . BACKGROUND : Neuronal loss in multiple sclerosis ( MS ) and its animal model , experimental autoimmune encephalomyelitis ( EAE ) , correlates with permanent neurological dysfunction . Current MS therapies have limited the ability to prevent neuronal damage . METHODS : We examined whether oral therapy with DB05073 , a pharmaceutical grade formulation of resveratrol , reduces neuronal loss during relapsing-remitting EAE . DB02709 activates Q96EB6 , an NAD+-dependent deacetylase that promotes mitochondrial function . RESULTS : Oral DB05073 prevented neuronal loss during optic neuritis , an inflammatory optic nerve lesion in MS and EAE . DB05073 also suppressed neurological dysfunction during EAE remission , and spinal cords from DB05073 -treated mice had significantly higher axonal density than vehicle-treated mice . Similar neuroprotection was mediated by SRT1720 , another Q96EB6 -activating compound ; and sirtinol , an Q96EB6 inhibitor , attenuated DB05073 neuroprotective effects . Q96EB6 activators did not prevent inflammation . CONCLUSIONS : These studies demonstrate that DB05073 attenuates neuronal damage and neurological dysfunction in EAE by a mechanism involving Q96EB6 activation . Q96EB6 activators are a potential oral therapy in MS . Microarray Analysis of the Major Depressive Disorder mRNA Profile Data . OBJECTIVE : Major depressive disorder ( MDD ) is a common mood disorder associated with several psychophysiological changes like disturbances of sleep , appetite , or sexual desire , and it affects the patients ' life seriously . We aimed to explore a genetic method to investigate the mechanism of MDD . METHODS : The mRNA expression profile ( GSE53987 ) of MDD was downloaded from Gene Expression Omnibus database , including 105 samples of three brain regions in post-mortem tissue suffered from MDD and unaffected controls . Differentially expressed genes ( DEGs ) in MDD were identified using the Limma package in R . Gene Ontology functions and Kyoto Enrichment of Genes and Genomes pathways of the selected DEGs were enriched using Database for Annotation , Visualization and Integrated Discovery . Protein-protein interactive network of DEGs was constructed using the Cytoscape software . RESULTS : Totally , 241 DEGs in MDD-hip group , 218 DEGs in MDD-pfc group , and 327 DEGs in MDD-str group were identified . Also , different kinds of biological processes of DEGs in each group were enriched . Besides , glycan biosynthesis of DEGs in MDD-str group , O95786 -like receptor signaling and pyrimidine metabolism of DEGs in the MDD-hip group were enriched , respectively . Moreover , several DEGs like Q05397 , Q13569 and P41208 in MDD-str group , P40126 , AR and P30968 in MDD-pfc group , and P31749 and P51617 in MDD-hip group were selected from PPI network . CONCLUSION : Our data suggests that the brain striatum tissue may be greatly affected by MDD , and DEGs like Q05397 , Q10471 and Q10471 in striatum , AR in prefrontal cortex and P51617 and P29459 in hippocampus may provide novel therapeutic basis for MDD treatment . Anticancer drug discovery targeting DNA hypermethylation . DNA methyltransferases ( DNMTs ) are important regulators of gene transcription and their roles in carcinogenesis have been a topic of considerable interest in the last few years . Diverse classes of chemical compounds including nucleotide analogues , adenosine analogues , aminobenzoic derivatives , polyphenols , hydrazines , phthalides , disulfides and antisenses are being discovered and evaluated as P26358 inhibitors targeting DNA hypermethylation . Among them , DB00928 5 and Decitabine 6 were launched recently . Several other compounds are under clinical trials . Some of these compounds were discovered from structure-based drug design . These compounds exert their DNA methylation inhibitory by different mechanisms . This review will present a brief account of various DNA methyltransferases and their biological functions , with focus on actuality of design and synthesis of various inhibitors of DNA hypermethylation as anticancer drugs . PPARγ regulates the mitochondrial dysfunction in human neural stem cells with tumor necrosis factor alpha . P37231 ( PPARγ ) belongs to a family of ligand-activated transcription factors , and its ligands are known to control many physiological and pathological conditions . The hypothesis of our study was that the PPARγ agonist ( rosiglitazone ) could mediate tumor necrosis factor alpha ( TNFα ) related to the regulation of human neural stem cells ( hNSCs ) , by which TNFα possibly fulfills important roles in neuronal impairment . The results show that PPARγ mediates the cell viability of hNSCs via the downregulation of the activity of caspase 3 , indicating that this rescue effect of PPARγ could improve the reduced levels of two mitochondrial regulators , adenosine monophosphate-activated protein kinase ( AMPK ) and Sirtuin 1 ( Q96EB6 ) in the hNSCs with TNFα . The stimulation of mitochondrial function by PPARγ was associated with activation of the Q07869 coactivator1 alpha ( PGC1α ) pathway by up-regulation of oxidative defense and mitochondrial systems . The above protective effects appeared to be exerted by a direct activation of the rosiglitazone , because it protected hNSCs from TNFα-evoked oxidative stress and mitochondrial deficiency . Here we show that the rosiglitazone protects hNSCs against Aβ-induced apoptosis and promotes cell survival . These findings extend our understanding of the central role of PPARγ in TNFα-related neuronal impairment , which probably increases risks of neurodegenerative diseases . The anti-inflammatory effects of PPARγ in the hNSCs with TNFα , and the involved mechanisms were also characterized . In silico identification of potent pancreatic triacylglycerol lipase inhibitors from traditional Chinese medicine . P16233 ( P16233 ) are primary lipases that are critical for triacylglyceride digestion in human . Since reduced metabolism of triacylglyceride might be a plausible concept for weight loss , we screened for potential P16233 inhibitors from traditional Chinese medicine ( TCM ) with the aim to identify weight loss candidate compounds . TCM candidates Aurantiamide , Cnidiadin , and 2-hexadecenoic acid exhibited higher Dock Scores than the commercial drug DB01083 , and were also predicted to have inhibitory characteristics against P16233 using constructed P08235 ( R(2) = 0.8664 ) and SVM ( R(2) = 0.9030 ) models . Molecular dynamics indicated that the TCM- P16233 complexes formed were stable . We identified that the P16233 binding site has several residues that can serve as anchors , and a hydrophobic corridor that provides additional stability to the complex . Aurantiamide , Cnidiadin , and 2-hexadecenoic acid all have features that correspond to these binding site features , indicating their potential as candidates for P16233 inhibitors . The information presented in this study may provide helpful insights to designing novel weight-control drugs . Complement inhibition and statins prevent fetal brain cortical abnormalities in a mouse model of preterm birth . Premature babies are particularly vulnerable to brain injury . In this study we focus on cortical brain damage associated with long-term cognitive , behavioral , attentional or socialization deficits in children born preterm . Using a mouse model of preterm birth ( PTB ) , we demonstrated that complement component C5a contributes to fetal cortical brain injury . Disruption of cortical dendritic and axonal cytoarchitecture was observed in PTB-mice . Fetuses deficient in P21730 ( -/- ) did not show cortical brain damage . Treatment with antibody anti- P01031 , that prevents generation of C5a , also prevented cortical fetal brain injury in PTB-mice . C5a also showed a detrimental effect on fetal cortical neuron development and survival in vitro . Increased glutamate release was observed in cortical neurons in culture exposed to C5a . Blockade of P21730 prevented glutamate increase and restored neurons dendritic and axonal growth and survival . Similarly , increased glutamate levels - measured by (1)HMRS - were observed in vivo in PTB-fetuses compared to age-matched controls . The blockade of glutamate receptors prevented C5a-induced abnormal growth and increased cell death in isolated fetal cortical neurons . Simvastatin and pravastatin prevented cortical fetal brain developmental and metabolic abnormalities -in vivo and in vitro . Neuroprotective effects of statins were mediated by Akt/ P31749 signaling pathways . This study shows that complement activation plays a crucial role in cortical fetal brain injury in P16233 and suggests that complement inhibitors and statins might be good therapeutic options to improve neonatal outcomes in preterm birth . Tyrosine dephosphorylation of P40763 in P49591 coronavirus-infected Vero E6 cells . Severe acute respiratory syndrome ( P49591 ) has become a global public health emergency . p38 mitogen-activated protein kinase ( MAPK ) and its downstream targets are activated in P49591 coronavirus ( P49591 -CoV ) -infected Vero E6 cells and activation of p38 MAPK enhances the cytopathic effects of P49591 -CoV infection . In addition , weak activation of Akt can not prevent P49591 -CoV infection-induced apoptosis in Vero E6 cells . In the present study , we demonstrated that signal transducer and activator of transcription ( P35610 ) 3 , which is constitutively phosphorylated at tyrosine ( DB00135 ) -705 and slightly phosphorylated at serine ( DB00133 ) -727 in Vero E6 cells , was dephosphorylated at DB00135 -705 on P49591 -CoV infection . In addition to phosphorylation of p38 MAPK in virus-infected cells , other MAPKs , i.e. , extracellular signal-regulated kinase ( P29323 ) 1/2 and c-Jun N-terminal kinase ( JNK ) , were phosphorylated . Although inhibitors of P27361 /2 and JNK ( PD98059 and SP600125 ) had no effect on phosphorylation status of P40763 , inhibitors of p38 MAPK ( SB203580 and SB202190 ) partially inhibited dephosphorylation of P40763 at DB00135 -705 . DB00135 -705-phosphorylated P40763 was localized mainly in the nucleus in mock infected cells , whereas P40763 disappeared from the nucleus in virus-infected cells . As P40763 acts as an activator of transcription in the nucleus , these results suggest that P40763 lacks its activity on transcription in P49591 -CoV-infected Vero E6 cells . The effect of feeding system in the expression of genes related with fat metabolism in semitendinous muscle in sheep . The effect of feeding system on the expression of P06858 , Q13085 , P49327 , P15090 , O75907 , O00767 , Q92523 , P54646 , P41159 , P36956 , P37231 , Q07869 and P17676 genes in semitendinous muscle was studied . Forty-four single born male lambs of the Rasa Aragonesa breed , allocated to four different dietary treatments , were used : grazing alfalfa , grazing alfalfa with supplement for lambs , indoor lambs with grazing ewes and drylot . Significant differences were found in the expression of genes P06858 , Q13085 , P49327 , P15090 , Q92523 and O00767 . Genes related to adipogenesis ( P06858 , Q13085 , P49327 , P15090 , and O00767 ) are up-regulated in the intensive groups . In grazing groups Q92523 gene expression , related to β-oxidation process , is up-regulated . The relative expression of Q92523 was 1.54 fold higher in Q9UNN4 +S , and 0.43 and 0.37 fold lower in IND- GRE and IND , respectively . The results support the hypothesis that changes in fatty acid profile due to feeding system implicate changes in the mRNA expression level of genes related with fat metabolism . Feeding strategy is an important tool to manipulate intramuscular fatty acid profile in meat through altering gene expression of enzymes related with fat metabolism . Concerted activation of ETS protein P50549 by P52701 coactivators , the acetyltransferase p300 and the receptor tyrosine kinase P04626 /Neu . Activator of thyroid and retinoic acid receptor ( Q9Y6Q9 ) is overexpressed in approximately 60 % of primary human breast tumors and belongs to the P52701 steroid receptor coactivator family . In this study , we identified a novel interaction partner of Q9Y6Q9 , the ETS transcription factor P50549 that is also heavily implicated in mammary tumor formation . Q9Y6Q9 and related P52701 family members ( steroid receptor coactivator-1 and glucocorticoid receptor-interacting protein-1 ( Q9Y3R0 ) ) augment P50549 -mediated transcription . Although Q9Y6Q9 and Q9Y3R0 can acetylate P50549 , this posttranslational modification of P50549 is not required for its stimulation by Q9Y6Q9 or Q9Y3R0 . In addition , Q9Y6Q9 collaborates with the p300 coactivator , a joint interaction partner of Q9Y6Q9 and P50549 , to stimulate P50549 function and the ability of p300 to acetylate P50549 is indispensable for this collaboration . Furthermore , the receptor tyrosine kinase P04626 /Neu , an oncoprotein particularly found overexpressed in breast tumors , cooperates with both Q9Y6Q9 and p300 to stimulate P50549 -mediated transcription . Thus , oncogenic P04626 /Neu and Q9Y6Q9 may synergize to orchestrate mammary tumorigenesis through the dysregulation of the transcription factor P50549 and its target genes . Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation : a novel role of the cysteinyl LT-1 receptor . Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues . Leukotrienes are potent inflammatory mediators , and similar to prostaglandins , are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues . Here we tested the hypothesis that blocking the negative regulation of leukotrienes , would lead to enhanced endochondral bone formation during fracture repair . A closed femoral fracture was created in mice . Animals were divided into three groups for treatment with either montelukast sodium , a cysteinyl leukotriene type 1 receptor antagonist ( trade name Singulair ) , zileuton , a P09917 enzyme inhibitor ( trade name DB00744 ) , or carrier alone . The fractures were analyzed using radiographs , quantitative gene expression , histology and histomorphometry , and immunohistochemistry . Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls . Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7 . Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation , and increased early bone formation markers when compared with controls . Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor , leading to increased chondrocyte proliferation at early time points . These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation , with important and previously unrecognized implications for both fracture repair , and in a broader context , systemic chondrocyte growth and differentiation . Systems pharmacology dissection of the anti-inflammatory mechanism for the medicinal herb Folium eriobotryae . Inflammation is a hallmark of many diseases like diabetes , cancers , atherosclerosis and arthritis . Thus , lots of concerns have been raised toward developing novel anti-inflammatory agents . Many alternative herbal medicines possess excellent anti-inflammatory properties , yet their precise mechanisms of action are yet to be elucidated . Here , a novel systems pharmacology approach based on a large number of chemical , biological and pharmacological data was developed and exemplified by a probe herb Folium Eriobotryae , a widely used clinical anti-inflammatory botanic drug . The results show that 11 ingredients of this herb with favorable pharmacokinetic properties are predicted as active compounds for anti-inflammatory treatment . In addition , via systematic network analyses , their targets are identified to be 43 inflammation-associated proteins including especially P35354 , P09917 , P37231 , P01375 and Q04206 that are mainly involved in the mitogen-activated protein kinase ( MAPK ) signaling pathway , the rheumatoid arthritis pathway and NF-κB signaling pathway . All these demonstrate that the integrated systems pharmacology method provides not only an effective tool to illustrate the anti-inflammatory mechanisms of herbs , but also a new systems-based approach for drug discovery from , but not limited to , herbs , especially when combined with further experimental validations . 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 . 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 . The influence of mast cell mediators on migration of SW756 cervical carcinoma cells . The role of mast cell mediators on cervical cancer cell migration was assessed using an in vitro assay of scratch wound healing onto monolayers of HPV18-positive cervical carcinoma cells ( SW756 ) . Migration of SW756 cells was accelerated by co-culture with the mast cell line LAD2 . This effect was inhibited by the P35367 antagonist DB06691 and the cannabinoid agonists 2-arachidonylglycerol ( 2AG ) and Win 55,212-2 . Therefore , the specific effects of histamine and cannabinoids on SW756 migration and LAD2 activation were analyzed . DB11320 added to the in vitro assay of scratch wound healing either increased or inhibited SW756 migration rate by acting either on P35367 or Q9H3N8 , respectively . Cannabinoids acted on P21554 receptors to inhibit SW756 migration . Supernatants from SW756 cells stimulated LAD2 cell degranulation , which in turn was inhibited by cannabinoids acting via CB2 receptors . RT-PCR showed that SW756 expressed mRNA for P21554 , CB2 , P35367 , P25021 , and Q9H3N8 . On the other hand , LAD2 expressed mRNA for all four HRs and CB2 . The results suggest that mast cells could be contributing to cervical cancer cell invasion and spreading by the release of histamine and cannabinoids . Therefore , therapeutic modulation of specific mast cell mediators may be beneficial for cervical cancer treatment . 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 . P49327 inhibitor orlistat induces apoptosis in T cell lymphoma : role of cell survival regulatory molecules . BACKGROUND : De novo fatty acid synthesis catalyzed by fatty acid synthase ( P49327 ) is crucial for tumor cell survival . Thus therapeutic targeting of P49327 is considered as a novel antineoplastic strategy . However , little is understood in this respect regarding malignancies of hematological origin . The present investigation was therefore , undertaken to study the molecular mechanisms of the antitumor action of P49327 inhibitor orlistat ( tetrahydrolipstatin ) using a murine model of a T cell lymphoma . METHODS : The antitumor efficacy of orlistat was investigated in vitro by estimating cell survival by MTT assay and apoptosis by Wright Giemsa , TUNEL , Annexin-V/PI staining and % DNA fragmentation . Generation of reactive oxygen species ( ROS ) in tumor cells was studied using fluorescence microscopy . Expression of genes and proteins was carried out by RT-PCR and western blot analyses respectively . P49327 and CPT-1 activity was estimated by spectrophotometer . Cytokines expression was analyzed by ELISA . RESULTS : We report that inhibition of P49327 with its specific inhibitor orlistat manifests tumor-specific inhibition of cell survival , accompanied by induction of apoptosis . DB01083 -treated tumor cells showed an altered ROS generation , shift in cytokine balance and modulated expression of cell survival regulatory molecules like HSP70 , Bcl2 , p53 , PUMA , P42574 and CAD . It was observed that IFN-γ mediates orlistat-dependent modulation of P49327 expression . CONCLUSION AND GENERAL SIGNIFICANCE : In this study , we report some of the so far unexplored novel aspects underlying the molecular mechanisms associated with orlistat-dependent modulation of tumor cell survival . These observations will help in designing antineoplastic therapeutic protocols using orlistat against malignancies of hematological origin . Intracellular P05067 Domain Regulates DB00133 - DB03381 - DB01992 Transferase Expression and Is Affected in Alzheimer 's Disease . Lipids play an important role as risk or protective factors in Alzheimer 's disease ( AD ) , a disease biochemically characterized by the accumulation of amyloid beta peptides ( Aβ ) , released by proteolytic processing of the amyloid precursor protein ( P05067 ) . Changes in sphingolipid metabolism have been associated to the development of AD . The key enzyme in sphingolipid de novo synthesis is serine-palmitoyl- DB01992 transferase ( P21549 ) . In the present study we identified a new physiological function of P05067 in sphingolipid synthesis . The P05067 intracellular domain ( AICD ) was found to decrease the expression of the P21549 subunit O15270 , the catalytic subunit of the P21549 heterodimer , resulting in that decreased P21549 activity . AICD function was dependent on Fe65 and O15270 levels are increased in P05067 knock-in mice missing a functional AICD domain . O15270 levels are also increased in familial and sporadic AD postmortem brains , suggesting that P21549 is involved in AD pathology . Effects of DB00482 and DB00744 on liver metastasis and lipidperoxidation in pancreatic cancer in Syrian hamsters . Selective inhibition of eicosanoid synthesis is thought to have effects on carcinogenesis in lung and colon cancer . However , it is still unknown whether pancreatic cancer might also be influenced . Therefore we evaluated the impact of selective cyclooxygenase-2 inhibitor DB00482 and selective P09917 inhibitor DB00744 on liver metastasis in a solid model of pancreatic adenocarcinoma in Syrian hamster . In week 33 , the animals were sacrificed and incidence of pancreatic carcinomas and number and size of liver metastases were determined . Activities of antioxidative enzymes ( GSHPX/SOD ) and concentrations of products of lipidperoxidation were measured in liver metastases and non-metastatic hepatic tissue . The incidence ( 54.5 vs. 100 % ) , number ( 3.17 +/- 0.98 vs. 6.75 +/- 0.71 ) and size ( 2.67 +/- 1.97 vs. 11.75 +/- 1.98 mm2 ) of liver metastases were decreased by combined therapy of DB00744 and DB00482 ( P < 0.05 ) . Furthermore , activities of GSHPX ( [ 73.77 +/- 5.67 ] *10(5) vs. [ 15.49 +/- 4.02 ] *10(5) U/mg prot. ; P < 0.05 ) and SOD ( 474.92 +/- 108.8 vs. 127.89 +/- 38.75 U/mg prot. ; P < 0.05 ) were increased , while lipidperoxidation ( 0.31 +/- 0.08 nmol/mg prot. vs. 1.54 +/- 0.55 nmol/mg prot. ; P < 0.05 ) was decreased by combination therapy , in non-metastatic hepatic tissue . Moreover , combined therapy increased lipidperoxidation in liver metastases ( 0.47 +/- 0.09 vs. 1.95 +/- 0.12 nmol/mg prot. ; P < 0.05 ) . Thus , a combination of DB00482 and DB00744 might be a new concept to decrease tumour growth in liver metastases in advanced pancreatic cancer . [ Molecular mechanism of age-related osteoporosis ] . Bone loss by ageing has been investigated from standpoints of systemic abnormality and some deficiency in osteoblastic bone formation . This seminar summarizes the involvements of a key molecule of adipocytic differentiation P37231 , essential P05019 signaling molecules P35568 and Q9Y4H2 , and an anti-aging gene klotho in the pathophysiology of age-related osteoporosis . Functional genomics identifies a mechanism for estrogen activation of the retinoic acid receptor alpha1 gene in breast cancer cells . The identification of estrogen receptor ( ERalpha ) target genes is crucial to our understanding of its predominant role in breast cancer . In this study , we used a chromatin immunoprecipitation ( ChIP ) -cloning strategy to identify ERalpha-regulatory modules and associated target genes in the human breast cancer cell line MCF-7 . We isolated 12 transcriptionally active genomic modules that recruit ERalpha and the coactivator steroid receptor coactivator ( P12931 ) -3 to different intensities in vivo . One of the ERalpha-regulatory modules identified is located 3.7 kb downstream of the first transcriptional start site of the P10276 locus , which encodes retinoic acid receptor alpha1 ( RARalpha1 ) . This module , which includes an estrogen response element ( ERE ) , is conserved between the human and mouse genomes . Direct binding of ERalpha to the ERE was shown using EMSAs , and transient transfections in MCF-7 cells demonstrated that endogenous ERalpha can induce estrogen-dependent transcriptional activation from the module or the ERE linked to a heterologous promoter . Furthermore , ChIP assays showed that the coregulators Q15788 , Q9Y6Q9 , and receptor-interacting protein 140 are recruited to this intronic module in an estrogen-dependent manner . As expected from previous studies , the transcription factor Sp1 can be detected at the P10276 alpha1 promoter by ChIP . However , treatment with estradiol did not influence Sp1 recruitment nor help recruit ERalpha to the promoter . Finally , ablation of the intronic ERE was sufficient to abrogate the up-regulation of P10276 alpha1 promoter activity by estradiol . Thus , this study uncovered a mechanism by which ERalpha significantly activates RARalpha1 expression in breast cancer cells and exemplifies the utility of functional genomics strategies in identifying long-distance regulatory modules for nuclear receptors . 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 . 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 . DB05007 -- a multitargeted tyrosine kinase inhibitor : results of a phase II study in subjects with non-small cell lung cancer who have progressed after responding to treatment with either gefitinib or erlotinib . INTRODUCTION : Although patients with non-small cell lung cancer ( NSCLC ) whose tumors harbor epidermal growth factor receptor ( P00533 ) activating mutations commonly experience significant regressions when treated with erlotinib or gefitinib , they uniformly develop resistance to these agents . The secondary P00533 T790M mutation is found in 50 % of patients with acquired resistance . Herein , we studied DB05007 , an oral small molecule inhibitor of multiple receptor tyrosine kinases , including P00533 , P35968 , P04626 , and EphB4 , in NSCLC patients known or suspected of having tumors harboring T790M . METHODS : Eligible patients included those with relapsed or recurrent advanced NSCLC who progressed after ≥12 weeks of stable disease or response to erlotinib or gefitinib and/or those patients with a documented P00533 T790M . DB05007 300 mg was administered once daily . The primary end point was objective response rate . Pretreatment plasma samples were collected for mutation testing of circulating tumor DNA . RESULTS : Forty-one patients were enrolled ; 33 were evaluable for efficacy . One partial response was observed ( response rate 3 % and 90 % confidence interval , 0 % to 14 % ) . Of patients whose tumors harbored T790M , 67 % ( 8/12 ) had progression of disease as best response compared with 14 % ( 3/21 ) of those without this mutation . Plasma samples from 40 patients were available for mutation testing , 14 ( 35 % ) of which were found to have P00533 mutations . CONCLUSIONS : The 3 % response rate observed did not meet the prespecified threshold to recommend further study of DB05007 in patients who develop acquired resistance to erlotinib or gefitinib . Patients with T790M had a significantly worse progression-free survival . Inhibition of the T790M gatekeeper mutant of the epidermal growth factor receptor by EXEL-7647 . PURPOSE : Agents inhibiting the epidermal growth factor receptor ( P00533 ) have shown clinical benefit in a subset of non-small cell lung cancer patients expressing amplified or mutationally activated P00533 . However , responsive patients can relapse as a result of selection for P00533 gene mutations that confer resistance to DB00171 competitive P00533 inhibitors , such as erlotinib and gefitinib . We describe here the activity of EXEL-7647 ( DB05007 ) , a novel spectrum-selective kinase inhibitor with potent activity against the P01133 and vascular endothelial growth factor receptor tyrosine kinase families , against both wild-type ( WT ) and mutant P00533 in vitro and in vivo . EXPERIMENTAL DESIGN : The activity of P00533 inhibitors against WT and mutant EGFRs and their effect on downstream signal transduction was examined in cellular assays and in vivo using A431 and MDA-MB-231 ( WT P00533 ) and H1975 ( L858R and T790M mutant P00533 ) xenograft tumors . RESULTS : EXEL-7647 shows potent and long-lived inhibition of the WT P00533 in vivo . In addition , EXEL-7647 inhibits cellular proliferation and P00533 pathway activation in the erlotinib-resistant H1975 cell line that harbors a double mutation ( L858R and T790M ) in the P00533 gene . In vivo efficacy studies show that EXEL-7647 substantially inhibited the growth of H1975 xenograft tumors and reduced both tumor P00533 signaling and tumor vessel density . Additionally , EXEL-7647 , in contrast to erlotinib , substantially inhibited the growth and vascularization of MDA-MB-231 xenografts , a model which is more reliant on signaling through vascular endothelial growth factor receptors . CONCLUSIONS : These studies provide a preclinical basis for clinical trials of DB05007 in solid tumors and in patients bearing tumors that are resistant to existing P00533 -targeted therapies .

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MH_train_1518

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interacts_with DB00477?

multiple_choice

Subunit complementation of thymidylate synthase in Plasmodium falciparum bifunctional dihydrofolate reductase-thymidylate synthase . P04818 of Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) functions as a dimeric enzyme with extensive contact between the two TS domains . Structural data of PfDHFR-TS shows that the formation of the two TS active sites involves contribution of the amino acid residues from both TS domains . DB00125 -470 donated from the adjoining domain is shown to hydrogen-bond to DB03800 , while DB00151 -490 is a key nucleophile for TS catalysis by attacking C-6 of DB03800 . However , mutants of the two series could complement one another , giving rise to active enzyme . By means of subunit complementation assay using DB00125 -470 and DB00151 -490 mutants , it is shown that co-transformants of both TS-inactive DB00125 -470 and DB00151 -490 mutants can complement the growth of thymidine auxotroph chi2913RecA(DE3) by formation of a functional TS heterodimer contributing from both DB00125 -470 and DB00151 -490 mutant subunits . 6-[3H]-FdUMP thymidylate synthase activity assay further elaborate the essence of restoration of TS activity . The TS k(cat) value of the R470D+C490A heterodimer is decreased by half from that of the wild-type PfDHFR-TS . However , the Km values for DB03800 and CH2H4folate of the R470D+C490A heterodimer are similar to those of wild-type enzyme , indicating that the catalytic efficiency of the functional TS from the R470D+C490A heterodimer is similar to the wild-type TS enzyme in P. falciparum P00374 -TS . 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 . c-Src modulates estrogen-induced stress and apoptosis in estrogen-deprived breast cancer cells . The emergence of anti-estrogen resistance in breast cancer is an important clinical phenomenon affecting long-term survival in this disease . Identifying factors that convey cell survival in this setting may guide improvements in treatment . Estrogen ( E2 ) can induce apoptosis in breast cancer cells that have been selected for survival after E2 deprivation for long periods ( MCF-7:5C cells ) , but the mechanisms underlying E2-induced stress in this setting have not been elucidated . Here , we report that the c-Src kinase functions as a key adapter protein for the estrogen receptor ( ER , P03372 ) in its activation of stress responses induced by E2 in MCF-7:5C cells . E2 elevated phosphorylation of c-Src , which was blocked by 4-hydroxytamoxifen ( DB04468 ) , suggesting that E2 activated c-Src through the ER . We found that E2 activated the sensors of the unfolded protein response ( UPR ) , IRE1α ( O75460 ) and Q9NZJ5 kinase ( Q9NZJ5 ) , the latter of which phosphorylates eukaryotic translation initiation factor-2α ( eIF2α ) . E2 also dramatically increased reactive oxygen species production and upregulated expression of heme oxygenase P09601 ( P09601 ) , an indicator of oxidative stress , along with the central energy sensor kinase AMPK ( P54646 ) . Pharmacologic or RNA interference-mediated inhibition of c-Src abolished the phosphorylation of eIF2α and AMPK , blocked E2-induced ROS production , and inhibited E2-induced apoptosis . Together , our results establish that c-Src kinase mediates stresses generated by E2 in long-term E2-deprived cells that trigger apoptosis . This work offers a mechanistic rationale for a new approach in the treatment of endocrine-resistant breast cancer . Proangiogenesis action of the thyroid hormone analog 3,5-diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2- to 3-fold by either T(4) or T(3) at 10(-8)-10(-7) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5-diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T(3) and T(4) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T(4) and T(3) for a novel cell surface hormone receptor site on integrin alphavbeta3 . The thyroid hormone analogs DITPA , T(4) , and T(4)-agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 /2 signal transduction cascade . Additionally , a specific integrin alphavbeta3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta3 , and are MAPK dependent . Competitive interaction of seven in absentia homolog-1A and Ca2+/calmodulin with the cytoplasmic tail of group 1 metabotropic glutamate receptors . BACKGROUND : Group 1 metabotropic glutamate receptors ( Q13255 and P41594 ) are coupled to inositol trisphosphate/Ca2+ signaling via G proteins and play an important role in excitatory synaptic transmission . To explore the regulation of group 1 mGluR function , we applied the yeast two-hybrid system using the intracellular carboxy-terminal domain of group 1 mGluRs ( group 1 ct-mGluRs ) and attempted to identify novel protein-protein interactions of group 1 mGluRs . RESULTS : The two-hybrid screening revealed a specific interaction between group 1 ct-mGluRs and Siah-1A , the mammalian homolog of Drosophila seven in absentia which is involved in photoreceptor cell differentiation via the ubiquitin/proteasome-dependent mechanism . This interaction occurs within a homologous 27-28 amino acid stretch within group 1 ct-mGluRs and requires the latter two-thirds of Siah-1A . Following coexpression in COS-7 cells , myc-tagged Siah-1A was coimmunoprecipitated with the flag-tagged ct- Q13255 by anti-flag antibody . Furthermore , in vitro binding revealed that Siah-1A and Ca2+/calmodulin ( P62158 ) binding sites overlap , such that Siah-1A binding is competitively inhibited by P62158 in a Ca2+-dependent manner . CONCLUSIONS : The results demonstrate a direct interaction between group 1 mGluRs and Siah-1A and suggest a novel modulatory mechanism mediated by a competitive interaction between Ca2+/ P62158 and Siah-1A . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . 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 . 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 . 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 . Examination of ceramic restorative material interfacial debonding using acoustic emission and optical coherence tomography . OBJECTIVE : CAD/ P62158 ceramic restorative material is routinely bonded to tooth substrates using adhesive cement . This study investigates micro-crack growth and damage in the ceramic/dentin adhesive interface under fatigue shear testing monitored using the acoustic emission ( AE ) technique with optical coherence tomography ( O75051 ) . METHODS : Ceramic/dentin adhesive samples were prepared to measure the shear bond strength ( SBS ) under static load . Fatigue shear testing was performed using a modified ISO14801 method . Loads in the fatigue tests were applied at 80 % , 70 % , and 60 % of the SBS to monitor interface debonding . The AE technique was used to detect micro-crack signals in static and fatigue shear bond tests . RESULTS : The results showed that the average SBS value in the static tests was 10.61±2.23MPa ( mean±standard deviation ) . The average number of fatigue cycles in which ceramic/dentin interface damage was detected in 80 % , 70 % and 60 % of the SBS were 152 , 1962 and 9646 , respectively . The acoustic behavior varied according to the applied load level . Events were emitted during 60 % and 70 % fatigue tests . A good correlation was observed between crack location in O75051 images and the number of AE signal hits . SIGNIFICANCE : The AE technique and O75051 images employed in this study could potentially be used as a pre-clinical assessment tool to determine the integrity of cemented load bearing restored ceramic material . Sustainable cyclic load stresses in ceramic/dentin-bonded specimens were substantially lower than the measured SBS . Predicted S-N curve showed that the maximum endured load was 4.18MPa passing 10(6) fatigue cyclic . Luteinized unruptured follicle syndrome increased by inactive disease and selective cyclooxygenase 2 inhibitors in women with inflammatory arthropathies . OBJECTIVE : Administration of nonsteroidal antiinflammatory drugs ( NSAIDs ) may impair fertility . The occurrence of the luteinized unruptured follicle ( LUF ) syndrome was assessed in women with inflammatory arthropathies exposed to NSAIDs and compared to that in nonexposed women . METHODS : Fourteen patients with inflammatory rheumatic disease , 29 women with noninflammatory musculoskeletal conditions , and 449 women not exposed to NSAIDs were studied by intravaginal ultrasound monitoring for follicular development and ovulation in 1 or more menstrual cycles . Disease activity was assessed in inflammatory rheumatic disease . RESULTS : In 59 monitored cycles of patients with continuous NSAID exposure , 35.6 % of LUF syndromes occurred compared to 3.4 % of LUF syndromes in untreated women ( P < 0.001 ) . DB01628 was responsible for 75 % of LUF syndromes in patients exposed continuously , whereas diclofenac generated 15 % of LUF syndromes . An ibuprofen dosage of 1,600 mg/day did not induce LUF syndrome either at continuous periovulatory or discontinuous exposure . Interestingly , the frequency of LUF syndrome was 46.2 % in patients with inactive inflammatory disease compared to 15 % in patients with active disease ( P = 0.023 ) . DB01628 generated LUF syndrome in 94.2 % of the cases with inactive disease versus 28.6 % in patients with active disease ( P = 0.003 ) . CONCLUSION : NSAIDs increased the risk of the LUF syndrome , particularly in patients with inactive disease . The selective cyclooxygenase 2 ( P35354 ) inhibitor etoricoxib was a more potent inductor of LUF syndrome than nonselective P36551 inhibitors . Continuous periovulatory exposure to NSAIDs should be avoided when planning a pregnancy in patients with rheumatic diseases . 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 . Hypothalamic regulation of the adenohypophyseal-testicular axis in the male chick embryo . An antibody against luteinizing hormone-releasing hormone ( P01148 ) as well as naloxone , an opioid antagonist , were added to the chorioallantoic membrane ( P62158 ) of 11.5- and 14.5-day-old male chick embryos and plasma testosterone ( T ) concentrations were determined . This protocol was designed to demonstrate : ( 1 ) Whether P01148 is essential in the regulation of the adenohypophyseal-testicular axis in the male embryo and ( 2 ) if P01148 is operative in this unit 's function , are opiatergic pathways involved in the secretion of P01148 by the hypothalamus . Both anti- P01148 and naloxone lowered plasma T levels in 14.5-day-old embryos , but not 11.5-day-old embryos . This indicates that the hypothalamus , via P01148 , begins to regulate the pituitary-testicular unit at some time between Days 11.5 and 14.5 , i.e. , the hypothalamo-adenohypophyseal-testicular axis is established . The results also strongly suggest that the normal secretory pattern of P01148 is dependent upon opiatergic innervation of the hypothalamus at the same embryonic time . Aspects of quality of primary care provided by physicians certified in phytotherapy in Switzerland . BACKGROUND : Data on the use of phytotherapy in primary care are scarce and difficult to compare ( e.g. different health-care systems , study designs ) . OBJECTIVE : Are there differences in Switzerland regarding demographic data , practice structure , process of care and outcome/ treatment satisfaction between primary care physicians certified in phytotherapy ( P62158 ) and physicians performing conventional primary care ( COM ) and their patients ? MATERIAL AND METHODS : Subgroup analysis of the data of phytotherapy of an observational study ( 2 cross-sectional surveys with 3 questionnaires ) which was performed as part of a nationwide evaluation program on complementary medicine ( Q9NZJ5 ) . A descriptive analysis was used to compare data . RESULTS : In survey A , 20 P62158 and 191 COM physicians participated , of which 14 and 84 , respectively , continued for survey B and recruited at least 276 P62158 and 1,395 COM patients . Findings show that P62158 physicians had less technical equipment ( e.g. x-rays ) than COM physicians , their consultation time was 25 % longer , and they used more non-drug therapies . Whereas in the SF-36 no differences could be identified between the groups , the EUROPEP showed significant differences in favour of P62158 patients . CONCLUSIONS : Preliminary data of the comparison between P62158 and COM physicians indicate few differences in demographic and practice structure data . Yet , due to differences in the process of care P62158 patients showed better treatment satisfaction than COM patients . This is probably due to their doctors ' communicative qualities and patient-oriented skills . To which degree this might be triggered due to phyto-pharmacosemiotic aspects needs to be investigated in a future study . Inhibition of aldehyde dehydrogenase 2 activity enhances antimycin-induced rat cardiomyocytes apoptosis through activation of MAPK signaling pathway . Aldehyde dehydrogenase 2 ( P05091 ) , a mitochondrial-specific enzyme , has been proved to be involved in oxidative stress-induced cell apoptosis , while little is known in cardiomyocytes . This study was aimed at investigating the role of P05091 in antimycin A-induced cardiomyocytes apoptosis by suppressing P05091 activity with a specific P05091 inhibitor DB02115 . Antimycin A ( 40μg/ml ) was used to induce neonatal cardiomyocytes apoptosis . DB02115 ( 60μM ) effectively inhibited P05091 activity by 50 % without own effect on cell apoptosis , and significantly enhanced antimycin A-induced cardiomyocytes apoptosis from 33.5±4.4 to 56.5±6.4 % ( Hochest method , p < 0.05 ) , and from 57.9±1.9 to 74.0±11.9 % ( FACS , p < 0.05 ) . Phosphorylation of activated MAPK signaling pathway , including extracellular signal-regulated kinase ( P27361 /2 ) , c-Jun NH2-terminal kinase ( JNK ) and p38 was also increased in antimycin A and daidzin treated cardiomyocytes compared to the cells treated with antimycin A alone . These findings indicated that modifying mitochondrial P05091 activity/expression might be a potential therapeutic option on reducing oxidative insults induced cardiomyocytes apoptosis . 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 . DB00106 and other gonadotrophin-releasing hormone antagonists in prostate cancer . Hormonal therapy is the main recommended treatment for locally advanced and metastatic prostate cancer . DB00044 -releasing hormone ( P01148 ) agonists , such as buserelin , goserelin , leuprorelin and triptorelin , stimulate the pituitary 's gonadotrophin-releasing hormone ( DB00644 ) receptor , ultimately leading to its de-sensitization and subsequent reduction of LH and testosterone levels . However , this reduction is accompanied by a well described increase or ' surge ' in LH and testosterone levels , necessitating the concomitant administration of an antiandrogen to combat the potential effects of transient acceleration in cancer activity . Two pure DB00644 antagonists have been developed , abarelix and degarelix , that are devoid of any agonist effect on the P30968 and consequently do not result in testosterone flare . DB00106 was the first DB00644 antagonist to be developed and was approved by the USA Food and Drug Administration in 2004 for the initiation of hormonal castration in advanced or metastasizing hormone-dependent prostate carcinoma , when rapid androgen suppression is necessary . Clinical data on both abarelix and degarelix show that they can produce rapid and sustained decreases in testosterone to castrate levels without the need for co-administration of an antiandrogen , and with a very low complication rate in the short term . 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 . [ 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 .

MH_train_1519

MH_train_1519

MH_train_1519

interacts_with DB00863?

multiple_choice

A study of natural killer cell lectin-like receptor P04264 gene ( P26718 / P26718 ) region polymorphisms in a European population sample . Recent evidence has reinforced the belief in immunosurveillance as a powerful mechanism against cancer development . The natural killer ( NK ) cell has been recognized as a potent agent of cancer immunosurveillance . A Japanese cohort study correlated natural cytotoxic activity levels with subsequent cancer development and identified NK cell lectin-like receptor P04264 gene ( P26718 / P26718 ) polymorphisms as genetic markers of cancer predisposition . In the present study , we genotyped 82 reference cell lines and 388 newborn samples at 10 P26718 region variants by TaqMan ( ( R ) ) allelic discrimination assays and showed that the same polymorphisms occur at similar frequencies in Europeans . The same haplotype block that has been associated with lower natural cytotoxic activity also occurred with the highest frequency in our sample . We further detected evidence suggestive of natural selection at some of the loci analyzed and more importantly , sex specificity of this selection . It appeared that heterozygosity at loci forming a haplotype block was unfavorable for boys . Given the relevance of NK cells in fetal survival , this finding has potential implications in the study of genetics of maternofetal recognition . Our preliminary findings are of marginal statistical significance and should be replicated in a larger sample . We believe that our results will increase awareness of the involvement of P26718 in cancer immunosurveillance and possibly in prenatal selection . Transient postoperative vascular endothelial growth factor ( P15692 ) -neutralisation improves graft survival in corneas with partly regressed inflammatory neovascularisation . BACKGROUND : High-risk keratoplasties are usually performed after an uninflamed and quiescent interval in corneas with partly regressed blood and lymphatic vessels . We analysed whether the inhibition of post-keratoplasty revascularisation in mice with partly regressed corneal vessels ( " intermediate-risk " ) improves graft survival . METHODS : Three interrupted stromal sutures ( 11-0 ) in corneas of Balb/c mice ( 6-8 weeks old ) were placed for 6 weeks . Six months after suture removal , penetrating keratoplasty was performed with C57BL/6 donors . The treatment group received a vascular endothelial growth factor-A specific cytokine trap ( DB08885 ) intraperitoneally at days 0 , 4 , 7 and 14 after keratoplasty ( 25 mg/kg per mouse ; controls received equal amounts of Fc protein ) . Pathological haemangiogenesis and lymphangiogenesis prior to as well as 3 days or 8 weeks after keratoplasty and graft survival were analysed . RESULTS : Three days after keratoplasty corneal revascularisation was sufficiently reduced by DB08885 ( haem-vascularised areas 42.7 % reduction ; lymph-vascularised areas 54.7 % reduction ) . Survival proportions 8 weeks after keratoplasty were 36 % in the treatment group compared with 9 % in the control group ( n = 11 ; p < 0.05 ) . At that time no differences in haemangiogenesis or lymphangiogenesis were observed between the two groups . CONCLUSION : Early transient postoperative induction of haemangiogenesis and lymphangiogenesis and reformation of regressed corneal blood and lymphatic vessels are important for transplant rejections after " intermediate-risk " corneal transplantation . Inhibition of drug-resistant mutants of P00519 , P10721 , and P01133 receptor kinases . To realize the full potential of targeted protein kinase inhibitors for the treatment of cancer , it is important to address the emergence of drug resistance in treated patients . Mutant forms of P11274 - P00519 , P10721 , and the P01133 receptor ( P00533 ) have been found that confer resistance to the drugs imatinib , gefitinib , and erlotinib . The mutations weaken or prevent drug binding , and interestingly , one of the most common sites of mutation in all three kinases is a highly conserved " gatekeeper " threonine residue near the kinase active site . We have identified existing clinical compounds that bind and inhibit drug-resistant mutant variants of P00519 , P10721 , and P00533 . We found that the Aurora kinase inhibitor VX-680 and the p38 inhibitor BIRB-796 inhibit the imatinib- and BMS-354825-resistant P00519 (T315I) kinase . The P10721 / P36888 inhibitor SU-11248 potently inhibits the imatinib-resistant P10721 (V559D/T670I) kinase , consistent with the clinical efficacy of SU-11248 against imatinib-resistant gastrointestinal tumors , and the P00533 inhibitors Q9Y259 -569 and DB05424 , but not GW-572016 and ZD-6474 , potently inhibit the gefitinib- and erlotinib-resistant P00533 (L858R/T790M) kinase . Q9Y259 -569 and DB05424 are already in clinical trials , and our results suggest that they should be considered for testing in the treatment of gefitinib/erlotinib-resistant non-small cell lung cancer . The results highlight the strategy of screening existing clinical compounds against newly identified drug-resistant mutant variants to find compounds that may serve as starting points for the development of next-generation drugs , or that could be used directly to treat patients that have acquired resistance to first-generation targeted therapy . The use of IRES-based bicistronic vectors allows the stable expression of recombinant G-protein coupled receptors such as NPY5 and histamine 4 . Stable expression of G protein coupled receptors in cell lines is a crucial tool for the characterization of the molecular pharmacology of receptors and the screening for new antagonists . However , in some instances , many difficulties have been encountered to obtain stable cell lines expressing functional receptors . Here , we addressed the question of vector optimization to establish cell lines expressing the human neuropeptide Y receptor 5 ( Q15761 ) or histamine receptor 4 ( Q9H3N8 ) . We have compared bicistronic vectors containing viral or cellular internal ribosome entry sites ( IRES ) , co-expressing the receptor and the neomycine resistance gene from a single mRNA , to a bigenic vector containing two distinct promoters upstream each different genes . This study is the first one to validate the use of three cellular IRESs for long-term transgene expression . Our results demonstrate for both Q15761 and Q9H3N8 that the bicistronic vectors with EMCV , P15692 , FGF1A or P09038 IRES provide clones expressing functional receptors with yields between 25 % and 100 % . In contrast , the bigenic vector provided no functional clones , related to a low expression of Q15761 mRNA . The cell lines expressing active receptor were stable after more than 50 passages . These data indicate that IRES-based bicistronic vectors are particularly appropriate to establish cell clones expressing active G-coupled protein receptors with a high yield . In the case of NPY5 , it was a new way to produce such a stable cell line . Furthermore , the characteristics-presented herein-of this receptor pharmacological property are perfectly in line with those reported in the literature . Inhibitory effect of gabapentin on N-methyl-D-aspartate receptors expressed in Xenopus oocytes . BACKGROUND : DB00996 ( O75925 ) is a prescription drug used for the treatment of neuropathic and post-operative pain . However , the mechanism by which it exerts its analgesic action is not well understood . Because intrathecal administration of O75925 has been shown to exert antinociceptive effects in animal studies , we hypothesized that the spinal cord may be a plausible action site . METHODS : We examined the effects of O75925 on neurotransmitter-gated ion channels and G protein-coupled inwardly rectifying potassium ( GIRK ) channels distributed in the spinal cord and involved in pain modulation . Recombinant human Q9UHB4 / Q12879 N-methyl-D-aspartate ( DB01221 ) , alpha(1)beta(2)gamma(2S)gamma-aminobutyric acid type A ( GABA(A) ) or alpha(1) glycine receptors , or P48549 / P48051 channels were expressed in Xenopus laevis oocytes and the effects of O75925 ( 0.1-1000 microM ) on them were assessed using a two-electrode , voltage-clamp system . RESULTS : GABA(A) and glycine receptors and GIRK channels were not affected by O75925 , even at the highest concentrations . Conversely , DB01221 receptors were inhibited by O75925 in a concentration-dependent manner , with significant inhibition observed at 10 microM . At 30 microM , O75925 inhibited the glutamate-concentration response curve without changing the half-maximal effective concentration or the Hill coefficient , indicating a non-competitive inhibition . Glycine decreased the inhibitory effect in a concentration-dependent manner . CONCLUSIONS : These findings suggest that the inhibitory effect of O75925 on DB01221 receptors may play an important role in the antinociceptive property of O75925 ; however , it does not appear that GABA(A) and glycine receptors or GIRK channels contribute to the pharmacological properties of O75925 . 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 . 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 ) . DB04743 , a P35354 inhibitor , does not reduce lesion size or number in a nude mouse model of endometriosis . BACKGROUND : Women with endometriosis have elevated levels of cyclooxygenase-2 ( P35354 ) in peritoneal macrophages and endometriotic tissue . Inhibition of P35354 has been shown to reduce inflammation , angiogenesis and cellular proliferation . It may also downregulate aromatase activity in ectopic endometrial lesions . Ectopic endometrial establishment and growth are therefore likely to be suppressed in the presence of P35354 inhibitors . We hypothesized that P35354 inhibition would reduce the size and number of ectopic human endometrial lesions in a nude mouse model of endometriosis . METHODS : The selective P35354 inhibitor , nimesulide , was administered to estrogen-supplemented nude mice implanted with human endometrial tissue . Ten days after implantation , the number and size of ectopic endometrial lesions were evaluated and compared with lesions from a control group . Immunohistochemical assessment of vascular development and macrophage and myofibroblast infiltration in control and treated lesions was performed . RESULTS : There was no difference in the number or size of ectopic endometrial lesions in control and nimesulide-treated nude mice . DB04743 did not induce a visually identifiable difference in blood vessel development or macrophage or myofibroblast infiltration in nude mouse explants . CONCLUSION : The hypothesized biological properties of P35354 inhibition did not influence lesion number or size in the nude mouse model of endometriosis . 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 . Halogen versus high-intensity light-curing of uncoated and pre-coated brackets : a shear bond strength study . AIM : To evaluate the shear bond strengths of adhesive pre-coated brackets ( P25054 ) and conventional uncoated brackets ( Victory ) cured with two different light-curing units : a conventional halogen light ( Visilux 2 ) and a micro-xenon light ( Aurys ) . SETTING : Ex vivo study MATERIALS AND METHODS : Sixty freshly extracted bovine permanent mandibular incisors were randomly assigned to one of four groups , each group consisting of 15 specimens . Two groups ( one for each type of bracket ) were exposed to the halogen light for 20 seconds and used as controls . The remaining two groups were cured with the micro-xenon light for 2 seconds . After 24 hours , all samples were tested in a shear mode on an Instron Machine . Analysis was by two-way Q9UNW9 with Scheffé 's test for comparisons , Kaplan-Meier survival estimates , and Cox model . The Chi-square ( chi(2) ) test was used to determine significant differences in the Q9Y4X5 scores . RESULTS : The mean shear bond strength of the uncoated brackets cured with Visilux 2 was significantly higher than those of all the other groups tested . Both groups cured with Visilux 2 produced significantly higher mean shear bond strengths than those of the corresponding groups cured with Aurys . No statistically significant differences were found between the two groups cured with Aurys . CONCLUSIONS : Compared to halogen light-curing , the micro-xenon light enables the clinician to reduce significantly the curing time of both P25054 and uncoated brackets , and although significantly lower , their shear bond strengths may be clinically acceptable . Identification of genes upregulated in Q9UM73 -positive and P00533 / P01116 / Q9UM73 -negative lung adenocarcinomas . Activation of the P00533 , P01116 , and Q9UM73 oncogenes defines 3 different pathways of molecular pathogenesis in lung adenocarcinoma . However , many tumors lack activation of any pathway ( triple-negative lung adenocarcinomas ) posing a challenge for prognosis and treatment . Here , we report an extensive genome-wide expression profiling of 226 primary human stage I-II lung adenocarcinomas that elucidates molecular characteristics of tumors that harbor Q9UM73 mutations or that lack P00533 , P01116 , and Q9UM73 mutations , that is , triple-negative adenocarcinomas . One hundred and seventy-four genes were selected as being upregulated specifically in 79 lung adenocarcinomas without P00533 and P01116 mutations . Unsupervised clustering using a 174-gene signature , including Q9UM73 itself , classified these 2 groups of tumors into Q9UM73 -positive cases and 2 distinct groups of triple-negative cases ( groups A and B ) . Notably , group A triple-negative cases had a worse prognosis for relapse and death , compared with cases with P00533 , P01116 , or Q9UM73 mutations or group B triple-negative cases . In Q9UM73 -positive tumors , 30 genes , including Q9UM73 and Q12879 , were commonly overexpressed , whereas in group A triple-negative cases , 9 genes were commonly overexpressed , including a candidate diagnostic/therapeutic target Q5TB30 , that were determined to be critical for predicting a worse prognosis . Our findings are important because they provide a molecular basis of Q9UM73 -positive lung adenocarcinomas and triple-negative lung adenocarcinomas and further stratify more or less aggressive subgroups of triple-negative lung ADC , possibly helping identify patients who may gain the most benefit from adjuvant chemotherapy after surgical resection . 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 . Targeted disruption of the methionine synthase gene in mice . Alterations in homocysteine , methionine , folate , and/or B12 homeostasis have been associated with neural tube defects , cardiovascular disease , and cancer . Q99707 , one of only two mammalian enzymes known to require vitamin B12 as a cofactor , lies at the intersection of these metabolic pathways . This enzyme catalyzes the transfer of a methyl group from 5-methyl- DB00116 to homocysteine , generating DB00116 and methionine . Human patients with methionine synthase deficiency exhibit homocysteinemia , homocysteinuria , and hypomethioninemia . They suffer from megaloblastic anemia with or without some degree of neural dysfunction and mental retardation . To better study the pathophysiology of methionine synthase deficiency , we utilized gene-targeting technology to inactivate the methionine synthase gene in mice . On average , heterozygous knockout mice from an outbred background have slightly elevated plasma homocysteine and methionine compared to wild-type mice but seem to be otherwise indistinguishable . Homozygous knockout embryos survive through implantation but die soon thereafter . Nutritional supplementation during pregnancy was unable to rescue embryos that were completely deficient in methionine synthase . Whether any human patients with methionine synthase deficiency have a complete absence of enzyme activity is unclear . These results demonstrate the importance of this enzyme for early development in mice and suggest either that methionine synthase-deficient patients have residual methionine synthase activity or that humans have a compensatory mechanism that is absent in mice . Candidate pathway polymorphisms in one-carbon metabolism and risk of rectal tumor mutations . We examined candidate polymorphisms in genes involved in the folate-mediated , one-carbon metabolism pathway , P26358 1311V , P11586 R134K and R653Q , P42898 R594Q , Q99707 D919G , Q9UBK8 H595Y and I22M , P34896 L474F , P41440 H27R , and Q13569 G199S , and associations with rectal tumor characteristics . We hypothesized that these candidate genes would influence CpG Island Methylator Phenotype and potentially P01116 or P04637 tumors . Data from a population-based study of 747 rectal cases ( 593 with tumor markers ) and 956 controls were evaluated using generalized estimating equations . We observed an increased risk of P04637 tumor mutations in homozygous carriers of the P11586 134K allele ( 0R=2.0 , 95 % CI 1.2-3.1 , P- trend=0.02 ) . In the presence of low folate intake , the R134K variant was associated with increased risk of CIMP+ tumors ( OR=2.8 , 95 % CI 1.04-7.7 ) . The Q9UBK8 I22M variant genotype was associated with a modest increased risk of P04637 mutations ( OR=1.7 , 95 % CI 1.2-2.5 , P-trend=0.001 ) . Our findings offer limited support that polymorphisms in one-carbon metabolism genes influence rectal tumor phenotype , and that folate may interact with P11586 to alter CIMP+ risk . 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 role of natural killer cells in chronic myeloid leukemia . Chronic myeloid leukemia is a neoplasia resulting from a translocation between chromosomes 9 and 22 producing the P11274 - P00519 hybrid known as the Philadelphia chromosome ( Ph ) . In chronic myeloid leukemia a proliferation of malignant myeloid cells occurs in the bone marrow due to excessive tyrosine kinase activity . In order to maintain homeostasis , natural killer cells , by means of receptors , identify the major histocompatibility complex on the surface of tumor cells and subsequently induce apoptosis . The P26718 receptor in the natural killer cells recognizes the transmembrane proteins related to major histocompatibility complex class I chain-related genes A and B ( Q29983 and Q29980 ) , and it is by the interaction between P26718 and Q29983 that natural killer cells exert cytotoxic activity against chronic myeloid leukemia tumor cells . However , in the case of chronic exposure of the P26718 receptor , the Q29983 ligand releases soluble proteins called sMICA from the tumor cell surface , which negatively modulate P26718 and enable the tumor cells to avoid lysis mediated by the natural killer cells . Blocking the formation of sMICA may be an important antitumor strategy . Treatment using tyrosine kinase inhibitors induces modulation of NKG2DL expression , which could favor the activity of the natural killer cells . However this mechanism has not been fully described in chronic myeloid leukemia . In the present study , we analyze the role of natural killer cells to reduce proliferation and in the cellular death of tumor cells in chronic myeloid leukemia . 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 randomised , double-blind study comparing lumiracoxib with naproxen for acute musculoskeletal pain . BACKGROUND : Some selective cyclooxygenase-2 ( P35354 ) inhibitors have been shown to provide analgesic efficacy in patients with acute pain . AIM : To compare the efficacy and safety of the P35354 inhibitor lumiracoxib 400 mg once daily ( qd ) and naproxen 500 mg twice daily ( bid ) in patients with acute musculoskeletal pain caused by uncomplicated soft tissue injury . METHODS : This was a randomised , double-blind , parallel-group , non-inferiority study set in 39 primary care centres in the UK . Patients were randomised to lumiracoxib 400 mg qd or naproxen 500 mg bid and took the study medication for as long as they felt that it was needed , up to day 7 . The primary efficacy analysis was the sum of the pain intensity difference ( 0-100 mm visual analogue scale ) determined morning and evening over the first 5 days of treatment ( SPID-5 ) . RESULTS : The intention-to-treat population comprised 406 patients [ lumiracoxib 400 mg qd ( n = 207 ) ; naproxen 500 mg bid ( n = 199 ) ] . Both treatments were effective in reducing pain intensity over 5 days . The mean SPID-5 scores were 117.0 mm.day for lumiracoxib and 118.2 mm.day for naproxen [ the treatment difference based on adjusted means from the ANCOVA was -2.78 mm.day , 95 % confidence interval ( CI ) -17.4 , 11.9 ] . The lower margin of the 95 % CI was above the predetermined non-inferiority margin ( -50 mm.day ) for SPID-5 , indicating non-inferiority of lumiracoxib compared with naproxen . Both treatments were well tolerated . CONCLUSION : DB01283 400 mg qd is as effective as naproxen 500 mg bid for the management of moderate-to-severe acute musculoskeletal pain . 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 . 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 .

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interacts_with DB01114?

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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 . 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 . In vitro effects of E3040 , a dual inhibitor of P09917 and thromboxane A(2) synthetase , on eicosanoid production . In vitro pharmacological profiles of E3040 , 6-hydroxy-5 , 7-dimethyl-2-(methylamino)-4-(3-pyridylmethyl) benzothiazole were investigated . Against the P09917 activity of rat basophilic leukemia cells , E3040 and zileuton ( a P09917 inhibitor ) had an IC(50) of 0.23 and 0.93 microM , respectively . Against the thromboxane A(2) synthetase activity of human platelets , E3040 had an IC(50) of 0.01 microM , which was comparable to that of OKY-1581 ( sodium ( E ) -3- [ 4-(3-pyridylmethyl) phenyl ] -2-methylacrylate , a thromboxane A(2) synthetase inhibitor ) . Against cyclooxygenase activity of sheep seminal vesicles , E3040 showed no inhibition ( IC(50) , > 300 microM ) . Sulfasalazine and DB00244 , therapeutic drugs for inflammatory bowel disease , inhibited P09917 activity with an IC(50) of 293 and 970 microM , respectively . Sulfasalazine inhibited thromboxane A(2) synthetase activity with an IC(50) of 20 microM . In rat peritoneal leukocytes , E3040 inhibited leukotriene B(4) and thromboxane B(2) production with an IC(50) of 0.17 and 0.24 microM , respectively . E3040 inhibited leukotriene B(4) production in human neutrophils and thromboxane B(2) production in human platelets ( IC(50) of 0.21 and 0.09 microM , respectively ) . These results indicated that E3040 potently inhibited P09917 and thromboxane A(2) synthetase and blocked leukotriene B(4) and thromboxane B(2) production in rat peritoneal and human blood cells . 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 . [ Roles of histamine in the pathogenesis of bronchial asthma and reevaluation of the clinical usefulness of antihistamines ] . DB11320 has been reported to play an important role in pathogenesis of bronchial asthma . However , H1-blockers are not recommended as the first drug for asthma therapy in the guidelines . DB11320 may play various roles in allergic airway inflammation through the H1 receptor ( P35367 ) , P25021 , and Q9H3N8 in immune cells including T lymphocytes and dendritic cells . We therefore evaluated its role in allergic airway inflammation with the use of histamine-deficient mice . The results suggested that histamine plays a role in the prevention of goblet cell hyperplasia . Organic cation transporter-3 ( O75051 -3 ) is thought to be a transporter of histamine . Polymorphism of O75051 -3 { R120R ( T/C ) } was associated with the severity of asthma . Recently , it has been proposed that both asthma and allergic rhinitis should be treated as a single airway disease . Comorbidity of asthma and allergic rhinitis is very high ( 70-80 % ) and they share similar allergic inflammation . H1-blockers are recommended as first-line drugs to treat allergic rhinitis in the guidelines . Therefore H1-blockers are strongly recommended for patients with both asthma and allergic rhinitis . Biomarker analysis of neoadjuvant doxorubicin/cyclophosphamide followed by ixabepilone or Paclitaxel in early-stage breast cancer . PURPOSE : Predictive biomarkers offer the potential to improve the benefit:risk ratio of a therapeutic agent . DB04845 achieves comparable pathologic complete response ( pCR ) rates to other active drugs in the neoadjuvant setting . This phase II trial was designed to investigate potential biomarkers that differentiate response to this agent . EXPERIMENTAL DESIGN : Women with untreated , histologically confirmed primary invasive breast adenocarcinoma received neoadjuvant doxorubicin/cyclophosphamide , followed by 1:1 randomization to ixabepilone ( n = 148 ) or paclitaxel ( n = 147 ) . Rates of pCR were compared between treatment arms based on predefined biomarker sets : Q13509 , Q9Y6A5 , and CAPG gene expression , a 20- and 26-gene expression model , P08183 protein expression , and other potential markers of sensitivity . βIII-tubulin protein expression is reported separately but is referred to here for completeness . All patients underwent a core needle biopsy of the primary cancer for molecular marker analysis before chemotherapy . Gene expression profiling data were used for molecular subtyping . RESULTS : There was no significant difference in the rate of pCR in both treatment arms in βIII-tubulin-positive patients . Higher pCR rates were observed among βIII-tubulin-positive patients than in βIII-tubulin-negative patients . Furthermore , no correlation was evident between Q13509 , Q9Y6A5 , and CAPG gene expression , P08183 protein expression , multi-gene expression models , and the efficacy of ixabepilone or paclitaxel , even within the estrogen receptor-negative subset . CONCLUSION : These results indicate that βIII-tubulin protein and mRNA expression , P08183 protein expression , Q9Y6A5 and CAPG gene expression , and multigene expression models ( 20- and 26-gene ) are not predictive markers for differentiating treatment benefit between ixabepilone and paclitaxel in early-stage breast cancer . DB00051 induced pulmonary sarcoid reaction . Sarcoidosis is a multisystem granulomatous inflammatory disease of unknown etiology . There is evidence that Tumor Necrosis Factor alpha ( P01375 -α ) antagonists are useful in the treatment of advanced or refractory disease . However , sarcoidosis-like reaction has been reported with P01375 -α blockade in other inflammatory conditions . Here we report a case of sarcoid-like reaction in a patient with psoriatic arthritis shortly after initiation of adalimumab therapy . Stopping adalimumab and systemic anti-inflammatory therapy with corticosteroids resulted in resolution of pulmonary symptoms and chest radiographic findings . Though P01375 -α plays a critical role in pathogenesis of sarcoidosis , the development of sarcoid reaction with P01375 -α blockade is paradoxical and the mechanism of this response remains unknown . P01375 -α induced sarcoid-reaction could involve multiple organs . Its development with one agent does not preclude therapy with other P01375 -α blockers . DB00055 induction of P13500 in human endothelial cells : a possible role for endothelial cell nitric oxide synthase . Classically , activated protein C ( P25054 ) of the protein C/protein S anticoagulant pathway has functioned not only to inactivate the procoagulant factors Va and VIIIa but also to inhibit the activity of plasminogen activator inhibitor-1 ( P05121 ) . More recent data have suggested that the protein C/protein S pathway may serve as a physiological link between coagulation and inflammation . This P25054 pathway link was proposed because of observations showing that P25054 could both modulate the effects of cytokines and block neutrophil activation . As a further extension of the effect(s) of P25054 on cytokines , we found that P25054 , at the equivalent physiological protein C concentration of 4 microg/ml , significantly upregulated monocyte chemotactic protein-1 ( P13500 ) RNA in human umbilical vein endothelial cells ( HUVECs ) , as indicated by a ribonuclease protection assay ( RPA ) at 3 and 6 h with a return to near basal levels by 24 h . ELISA determinations demonstrated that 4 microg/ml of P25054 induced a significant ( P=.0001 ) increase in P13500 protein production over basal levels within a 24-h period . At the same concentration , P25054 downregulated endothelial cell nitric oxide synthase ( P29474 ) RNA . Downregulation first became apparent at 6 h and continued through 48 h of culture . This downregulation was concentration dependent over a range of 1.3-12 microg/ml , and there was no effect on cell viability within this range . In support of other studies , we also found that exogenously added nitric oxide ( NO ) inhibited P13500 production . These data suggest that P25054 may induce P13500 through the inhibition of P29474 . P05231 promotes head and neck tumor metastasis by inducing epithelial-mesenchymal transition via the JAK- P40763 -SNAIL signaling pathway . Epithelial-mesenchymal transition ( EMT ) is a key process in tumor metastatic cascade that is characterized by the loss of cell-cell junctions and cell polarity , resulting in the acquisition of migratory and invasive properties . However , the precise molecular events that initiate this complex EMT process in head and neck cancers are poorly understood . Increasing evidence suggests that tumor microenvironment plays an important role in promoting EMT in tumor cells . We have previously shown that head and neck tumors exhibit significantly higher Bcl-2 expression in tumor-associated endothelial cells and overexpression of Bcl-2 alone in tumor-associated endothelial cells was sufficient to enhance tumor metastasis of oral squamous cell carcinoma in a severe combined immunodeficient ( SCID ) mouse model . In this study , we show that endothelial cells expressing Bcl-2 ( EC-Bcl-2 ) , when cocultured with head and neck tumor cells ( CAL27 ) , significantly enhance EMT-related changes in tumor cells predominantly by the secretion of P05231 . Treatment with recombinant P05231 or stable P05231 overexpression in CAL27 cells or immortalized oral epithelial cells ( IOE ) significantly induced the expression of mesenchymal marker , vimentin , while repressing P12830 expression via the JAK/ P40763 /Snail signaling pathway . These EMT-related changes were further associated with enhanced tumor and IOE cell scattering and motility . P40763 knockdown significantly reversed P05231 -mediated tumor and IOE cell motility by inhibiting Q05397 activation . Furthermore , tumor cells overexpressing P05231 showed marked increase in lymph node and lung metastasis in a SCID mouse xenograft model . Taken together , these results show a novel function for P05231 in mediating EMT in head and neck tumor cells and increasing their metastatic potential . P35367 -stimulated interleukin 8 and granulocyte macrophage colony-stimulating factor production by bronchial epithelial cells requires extracellular signal-regulated kinase signaling via protein kinase C . BACKGROUND : DB11320 stimulates the release of several cytokines , such as interleukin ( IL ) -8 and granulocyte macrophage colony-stimulating factor , from bronchial epithelial cells . However , the functional individual histamine receptor subtype and intracellular signaling in bronchial epithelial cells are poorly defined . METHODS : Using human primary epithelial cells and the NCI-H292 cell line , we examined the expression of histamine receptor subtypes and histamine-induced second messenger . We also evaluated the involvements of mitogen-activated protein kinase , protein kinase C ( PKC ) and epidermal growth factor receptor in cytokine expression caused by histamine . RESULTS : P35367 ( P35367 ) was the only subtype expressed in both types of cells . DB11320 elevated intracellular calcium ion without affecting DB02527 levels . DB11320 induced the phosphorylation of extracellular signal-regulated kinase ( P29323 ) 1/2 . DB11320 also phosphorylated PKC and myristoylated alanine-rich C kinase substrate . Ro-31-8220 , a PKC inhibitor , and PD98059 , a mitogen-activated protein/ P29323 kinase inhibitor , suppressed the histamine-induced P29323 activation and the production of granulocyte macrophage colony-stimulating factor and P10145 . On the contrary , histamine had no effect on the phosphorylation of epidermal growth factor receptor , and its specific inhibitor AG1478 failed to inhibit the histamine-induced P29323 activation . Olopatadine , an H1 antagonist , completely blocked the histamine-related responses , whereas H2 and H3 antagonists did not . DB11320 also augmented the P10145 production caused by P05112 or tumor necrosis factor-alpha . CONCLUSIONS : The P35367 -PKC- P29323 pathway may play crucial roles in eliciting cytokine production from bronchial epithelial cells stimulated by histamine , leading to airway inflammation . New analogs of vitamin D3 . Calcitriol , the most active metabolite of vitamin D , controls parathyroid gland growth and suppresses the synthesis and secretion of parathyroid hormone ( PTH ) . However , because of its potent effects on intestinal calcium absorption and bone mobilization , calcitriol treatment can induce hypercalcemia , often precluding its use at therapeutic doses . Hyperphosphatemia is also a persistent problem among patients undergoing chronic hemodialysis and can be aggravated by therapeutic doses of calcitriol . Several pharmaceutical companies were able to modify the side-chain of the 1,25(OH)2D3 , allowing some of these new analogs to retain the action on the parathyroid glands while decreasing their hypercalcemic and hyperphosphatemic effects . The structure-activity relationship for ligand-mediated transcriptional regulation has been studied in detail . In some analogs the serum binding protein ( DBP ) plays a key role in determining the pharmacokinetics of the vitamin D compound . The affinity to DBP for 22-oxacalcitriol ( O75051 ) , an analog of calcitriol for the treatment of secondary hyperparathryoidism , is approximately 300-400 times lower than that of calcitriol and the analog is rapidly cleared from the circulation . The mechanisms for the selectivity of 19-nor-1,25(OH)2D2 ( paricalcitol ) ( DB00910 ) another analog of calcitriol , is clearly different from O75051 . Although the mechanisms of action is not completely known , it does appear that paricalcitol down-regulates the P11473 in the intestine . It is likely that the unique biological profiles of vitamin D analogs in vivo are due to multiple mechanisms . Understanding the molecular basis of the analog selectivity will not only provide an explanation for their unique actions but allow intelligent design of more effective analogs in the future . DB05812 acetate : a review of its use in patients with metastatic castration-resistant prostate cancer . DB05812 acetate ( Zytiga(®) ) is an orally administered , selective inhibitor of the 17α-hydroxylase and C17,20-lyase enzymatic activities of cytochrome P450 ( CYP ) 17 . P05093 is required for androgen biosynthesis , with androgen receptor signalling crucial in the progression from primary to metastatic prostate cancer . DB05812 acetate is approved in the European Union and the US , in combination with prednisone or prednisolone , for the treatment of men with metastatic castration-resistant prostate cancer ( CRPC ) . When administered in combination with prednisone in a placebo-controlled , multinational phase III study , abiraterone acetate significantly prolonged overall survival and radiographic progression-free survival ( rPFS ) in men with metastatic CRPC who had previously received docetaxel . In men with metastatic CRPC who had not previously received chemotherapy participating in a placebo-controlled , multinational phase III study , there was a strong trend towards an overall survival benefit , a significant prolongation in rPFS and significant delays in clinical decline , the need for chemotherapy and the onset of pain observed . Given the nature of the therapy , the overall tolerability profile of abiraterone acetate , in combination with prednisone , was acceptable in men with metastatic CRPC . DB05812 acetate is associated with hypokalaemia , hypertension , and fluid retention or oedema , secondary to its mechanism of action , and with cardiac adverse events and hepatotoxicity ; however , in the phase III studies the incidences of the most frequently reported grade 3 or 4 adverse events of special interest were relatively low . Although the final overall survival data in men with metastatic CRPC who have not previously received chemotherapy are awaited , current evidence indicates that abiraterone acetate is a useful option for the treatment of metastatic CRPC . Multipotent cancer stem cells derived from human malignant peritoneal mesothelioma promote tumorigenesis . During the progression of malignant peritoneal mesothelioma ( MPeM ) , tumor nodules propagate diffusely within the abdomen and tumors are characterized by distinct phenotypic sub-types . Recent studies in solid organ cancers have shown that cancer stem cells ( CSCs ) play a pivotal role in the initiation and progression of tumors . However , it is not known whether tumorigenic stem cells exist and whether they promote tumor growth in MPeM . In this study , we developed and characterized a CSC model for MPeM using stably expandable tumorigenic stem cells derived from patient tumors . We found morphologically distinct populations of CSCs that divide asymmetrically or symmetrically in MPeM in vitro cell culture . The MPeM stem cells ( MPeMSCs ) express stem cell markers c-MYC , P48681 and P35968 and in the presence of matrix components cells form colony spheres . MPeMSCs are multipotent , differentiate into neuronal , vascular and adipose progeny upon defined induction and the differentiating cells express lineage-specific markers such as Q13509 , an early neuronal marker ; P04275 , P15692 , P49767 and P10145 , endothelial markers ; and PPARγ and P15090 , adipose markers . Xenotransplantation experiments using MPeMSCs demonstrated early tumor growth compared with parental cells . Limiting dilution experiments using MPeMSCs and endothelial lineage-induced cells derived from a single MPeMSC resulted in early tumor growth in the latter group indicating that endothelial differentiation of MPeMSCs is important for MPeM tumor initiation . Our observation that the MPeM tumors contain stem cells with tumorigenic potential has important implications for understanding the cells of origin and tumor progression in MPeM and hence targeting CSCs may be a useful strategy to inhibit malignant progression . DB09302 : Q8NBP7 inhibitor for LDL cholesterol reduction . The proof of concept that proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) inhibition affects cholesterol levels was first established after the demonstration that Q8NBP7 loss-of-function mutations result in a significant drop in circulating LDL cholesterol levels . Subsequent studies revealed that Q8NBP7 binds the epidermal growth factor precursor homology domain-A on the surface LDL Receptor ( P01130 ) and directs P01130 and Q8NBP7 for lysosomal degradation . DB09302 ( also known as SAR236553/REGN727 ) is a monoclonal antibody that binds circulating Q8NBP7 and blocks its interactions with surface P01130 . DB09302 clinical trials with different doses on different administration schedules were shown to significantly reduce LDL cholesterol both as a mono-therapy and in combination with statins or ezetimibe . Although there is great potential for anti- Q8NBP7 therapies in the management of cholesterol metabolism , there is no clear evidence yet that blocking Q8NBP7 reduces cardiovascular disease outcome . This is being investigated in ongoing Phase III clinical trials with alirocumab . 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 . Comparison of the effects of firocoxib , carprofen and vedaprofen in a sodium urate crystal induced synovitis model of arthritis in dogs . A randomized , placebo-controlled , four-period cross-over laboratory study involving eight dogs was conducted to confirm the effective analgesic dose of firocoxib , a selective P35354 inhibitor , in a synovitis model of arthritis . DB09217 was compared to vedaprofen and carprofen , and the effect , defined as a change in weight bearing measured via peak ground reaction , was evaluated at treatment dose levels . A lameness score on a five point scale was also assigned to the affected limb . Peak vertical ground reaction force was considered to be the most relevant measurement in this study . The firocoxib treatment group performed significantly better than placebo at the 3 h post-treatment time point and significantly better than placebo and carprofen at the 7 h post-treatment time point . Improvement in lameness score was also significantly better in the dogs treated with firocoxib than placebo and carprofen at both the 3 and 7 h post-treatment time points . Selective inhibition of prostaglandin E2 receptors EP2 and EP4 inhibits adhesion of human endometriotic epithelial and stromal cells through suppression of integrin-mediated mechanisms . Endometriosis is a chronic gynecological disease of reproductive age women characterized by the presence of functional endometrial tissues outside the uterine cavity . Interactions between the endometriotic cells and the peritoneal extracellular matrix proteins ( Q13201 ) are crucial mechanisms that allow adhesion of the endometriotic cells into peritoneal mesothelia . DB00917 ( DB00917 ) plays an important role in the pathogenesis of endometriosis . In previous studies , we have reported that selective inhibition of DB00917 receptors PTGER2 and P35408 decreases survival and invasion of human endometriotic epithelial and stromal cells through multiple mechanisms . Results of the present study indicates that selective inhibition of PTGER2- and P35408 -mediated DB00917 signaling 1 ) decreases the expression and/or activity of specific integrin receptor subunits Itgb1 ( beta1 ) and Itgb3 ( beta3 ) but not Itgb5 ( beta5 ) , Itga1 ( alpha1 ) , Itga2 ( alpha2 ) , Itga5 ( alpha5 ) , and Itgav ( alphav ) ; 2 ) decreases integrin-signaling components focal adhesion kinase or protein kinase 2 ( Q05397 ) and talin proteins ; 3 ) inhibits interactions between Itgb1/Itgb3 subunits , Q05397 , and talin and PTGER2/ P35408 proteins through beta-arrestin-1 and Src kinase protein complex in human endometriotic epithelial cells 12Z and stromal cells 22B ; and 4 ) decreases adhesion of 12Z and 22B cells to Q13201 collagen I , collagen IV , fibronectin , and vitronectin in a substrate-specific manner . These novel findings provide an important molecular framework for further evaluation of selective inhibition of PTGER2 and P35408 as potential nonsteroidal therapy to expand the spectrum of currently available treatment options for endometriosis in child-bearing age women . Overproduced interleukin 6 decreases blood lipid levels via upregulation of very-low-density lipoprotein receptor . BACKGROUND : Interleukin 6 ( P05231 ) blockade raises blood lipid levels in patients with rheumatoid arthritis . OBJECTIVE : To examine the influence of P05231 on lipid metabolism . METHODS : Vascular smooth muscle cells ( VSMC ) were cultured in the presence of P05231 , soluble P05231 receptor ( sIL6R ) , P05231 +sIL6R or tumour necrosis factor alpha ( TNFalpha ) for 24 h . After culture , the expression of very-low-density lipoprotein receptor ( P98155 ) , low-density lipoprotein receptor ( P01130 ) and low-density lipoprotein-related protein-1 ( Q07954 ) were measured by real-time PCR . Human P05231 was injected into mice twice a day for 2 weeks and then P98155 expression in several tissues and the change of total cholesterol ( TC ) and triglyceride ( TG ) levels were investigated . Finally , the effect of anti- P05231 receptor ( P08887 ) antibody injection on blood lipid levels was examined . RESULTS : P05231 +sIL6R significantly induced expression of P98155 mRNA in VSMC ( 8.6-fold , p < 0.05 ) , but P05231 or sIL6R alone and TNFalpha did not do so . None of these cytokines induced P01130 and Q07954 mRNA expression . P05231 injection into mice increased the expression of P98155 in heart , adipose tissue and liver and decreased TC and TG levels . The injection of anti- P08887 antibody normalised the reduced levels of TC and TG caused by P05231 injection , whereas it had no influence on the levels of TC and TG in normal mice . CONCLUSIONS : Overproduced P05231 decreased blood lipid levels by increasing P98155 expression in several tissues . It is concluded that P05231 blockade normalises reduced lipid levels caused by P05231 , but does not affect normal lipid metabolism . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation .

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DB02709 , a polyphenol found in grapes , suppresses oxidative damage and stimulates apoptosis during early colonic inflammation in rats . Oxidative stress , neutrophil infiltration , proinflammatory cytokines and eicosanoid generation are clearly involved in the pathogenesis of intestinal bowel disease . DB02709 , a polyphenolic compound found in grapes and wine , has been shown to have anti-inflammatory , antioxidant , antitumour and immunomodulatory activities , however , its effects on experimental colitis remain unknown . We have investigated the effects of resveratrol on the colon injury caused by intracolonic instillation of trinitrobenzenesulphonic acid ( TNBS ) in rats . We determined the production of prostaglandin (PG)E(2) and P52209 (2) in colon mucosa and the expression of cyclo-oxygenases ( P36551 ) -1 and -2 immunohistochemically . The inflammatory response was assessed by histology and myeloperoxidase activity , as an index of neutrophil infiltration . P01584 production , histological and histochemical analysis of the lesions were also carried out . Finally , since resveratrol has been found to modulate apoptosis we intended to elucidate its effects on colonic mucosa under early acute inflammatory conditions . DB02709 ( 5-10mg/kg/day ) significantly reduced the degree of colonic injury , the index of neutrophil infiltration and the levels of the cytokine . DB02709 did not revert the increased PGE(2) levels but produced a significant fall in the P52209 (2) concentration . Compared with inflamed colon , no changes in staining for P23219 were observed in colon of resveratrol and TNBS-treated rats . In contrast , P35354 expression was decreased . Furthermore , resveratrol enhanced apoptosis compared with already high level induced by TNBS . In conclusion , resveratrol reduces the damage in experimentally induced colitis , alleviates the oxidative events and stimulates apoptosis . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . 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 . Superoxide dismutase is an abundant component in cell bodies , dendrites , and axons of motor neurons and in a subset of other neurons . Mutation in superoxide dismutase 1 ( P00441 ) , a Cu/Zn enzyme that removes oxygen radicals and protects against oxidative injury , has been implicated in some cases of familial amyotrophic lateral sclerosis ( FALS ) . As a first approach to examining the mechanism(s) through which these mutations cause specific degeneration of motor neurons , we have used immunocytochemistry to identify the distribution of P00441 in populations of cells in the peripheral and central nervous systems . In the spinal cord , intense P00441 immunoreactivity was present in motor neurons , interneurons , and substantia gelatinosa . In motor neurons , P00441 immunoreactivity was abundant in perikarya , dendrites , and axons ; most of this activity appeared to be free in the cytoplasm , although a portion was associated with membranous vesicles , presumably peroxisomes . Since a variety of central nervous system neurons , including pyramidal cells in cerebral cortex and neurons of the P07451 and P22748 sectors of the hippocampus , showed high immunoreactivity but are unaffected in P35858 , the apparent abundance of P00441 does not predict vulnerability of neurons to mutations in P00441 . Rather , P00441 accumulates in many neuronal populations but is particularly abundant in motor neurons . Consistent with recent studies of FALS-linked P00441 mutations in vitro and in transgenic mice , our findings offer further support for the view that the mutations confer a gain of adverse function . In this view , high , rather than limiting , levels of P00441 may place motor neurons selectively at risk in FALS . Interspecies hybrid tryptophan synthase-modified beta 2 protein formed from separate folding regions of the beta monomer . Escherichia coli and Serratia marcescens tryptophan synthase beta 2 protein ( EC 4.2.1.20 ) was subjected to mild trypsin proteolysis . Two separate folding regions ( domains ) of the E. coli ( EF1 and P13639 ) and the S. marcescens ( SF1 and Q07955 ) enzyme were shown to form interspecies hybrid reconstituted molecules [ (EF1- Q07955 )2 and (SF1- P13639 )2 ] and intraspecies reconstituted molecules [ (EF1- P13639 )2 and (SF1- Q07955 )2 ] with equal efficiency . The data suggest that structural regions , associated with beta monomer assembly , exist somewhere on the domain fragments and that these regions are conserved . 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 . SAR and in vivo evaluation of 4-aryl-2-aminoalkylpyrimidines as potent and selective O60674 ( O60674 ) inhibitors . We report the discovery of a series of 4-aryl-2-aminoalkylpyrimidine derivatives as potent and selective O60674 inhibitors . High throughput screening of our in-house compound library led to the identification of hit 1 , from which optimization resulted in the discovery of highly potent and selective O60674 inhibitors . Advanced lead 10d demonstrated a significant dose-dependent pharmacodynamic and antitumor effect in a mouse xenograft model . Based upon the desirable profile of 10d ( DB05243 ) it was advanced into clinical trials . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . [ DB00594 attenuates hypoxia-induced proliferation of rats pulmonary artery smooth muscle cells by suppressing Na+/ H+ exchanger-1 ] . AIM : To study the influence of Na+/H+ exchange inhibitor amiloride on hypoxia-induced proliferation in rats pulmonary artery smooth muscle cells ( PASMCs ) , also observe the change of Na+/H+ exchanger-1 ( P19634 ) activity and expression . METHODS : Rats PASMGs were cultured in normoxia ( 21 % O2 ) or hypoxia ( 2 % O2 ) for 24 hours , as well as administered amiloride with various concentrations , cultured for 24 hours , then determined MTT OD values and rates of P12004 positive cells to investigate cells proliferation , moreover intracellular pH was determined by interactive Laser Cytometer , and Na+/H+ exchanger-1 mRNA expression was determined by RT-PCR . RESULTS : Hypoxic exposure heightened intracellular pH and mRNA expression of P19634 in PASMCs , however , 3.123-50 micromol/L amiloride depressed them gradually . Additionally , hypoxic exposure raised MTT OD value and rates of P12004 positive cells , similarly , the above two indexes descended gradually with presence of 3.125-50 micromol/L amiloride . CONCLUSION : Na+/H+ exchange inhibitor amiloride can suppress hypoxia-induced proliferation in pulmonary artery smooth muscle cells , which is due to depress activity and expression of P19634 . Characterization of disease-associated mutations affecting an exonic splicing enhancer and two cryptic splice sites in exon 13 of the cystic fibrosis transmembrane conductance regulator gene . Sequences in exons can play an important role in constitutive and regulated pre-mRNA splicing . Since exonic splicing regulatory sequences are generally poorly conserved and their mechanism of action is not well understood , the consequence of exonic mutations on splicing can only be determined empirically . In this study , we have investigated the consequence of two cystic fibrosis ( CF ) disease-causing mutations , E656X and 2108delA , on the function of a putative exonic splicing enhancer ( ESE ) in exon 13 of the P13569 gene . We have also determined whether five other CF mutations D648V , D651N , G654S , E664X and T665S located near this putative ESE could lead to aberrant splicing of exon 13 . Using minigene constructs , we have demonstrated that the E656X and 2108delA mutations could indeed cause aberrant splicing in a predicted manner , supporting a role for the putative ESE sequence in pre-mRNA splicing . In addition , we have shown that D648V , E664X and T665S mutations could cause aberrant splicing of exon 13 by improving the polypyrimidine tracts of two cryptic 3' splice sites . We also provide evidence that the relative levels of two splicing factors , hTra2alpha and Q07955 /ASF , could alter the effect on splicing of some of the exon 13 disease mutations . Taken together , our results suggest that the severity of CF disease could be modulated by changes in the fidelity of P13569 pre-mRNA splicing . Down-regulated P13569 During Aging Contributes to Benign Prostatic Hyperplasia . Benign prostatic hyperplasia ( BPH ) is a hyper-proliferative disease of the aging prostate ; however , the exact mechanism underlying the development of BPH remains incompletely understood . The present study investigated the possible involvement of the cystic fibrosis transmembrane conductance regulator ( P13569 ) , which has been previously shown to negatively regulate nuclear factor-κB ( NF-κB ) /cyclooxygenase 2 ( P35354 ) /prostaglandin E2 ( DB00917 ) pathway , in the pathogenesis of BPH . Our results showed decreasing P13569 and increasing P35354 expression in rat prostate tissues with aging . Furthermore , suppression of P13569 led to increased expression of P35354 and over-production of DB00917 in a normal human prostate epithelial cell line ( PNT1A ) with elevated NF-κB activity . DB00917 stimulated the proliferation of primary rat prostate stromal cells but not epithelial cells , with increased P12004 expression . In addition , the condition medium from PNT1A cells after inhibition or knockdown of P13569 promoted cell proliferation of prostate stromal cells which could be reversed by P35354 or NF-κB inhibitor . More importantly , the involvement of P13569 in BPH was further demonstrated by the down-regulation of P13569 and up-regulation of P35354 /NF-κB in human BPH samples . The present results suggest that P13569 may be involved in regulating DB00917 production through its negative regulation on NF-κB/ P35354 pathway in prostate epithelial cells , which consequently stimulates cell growth of prostate stromal cells . The overstimulation of prostate stromal cell proliferation by down-regulation of P13569 -enhanced DB00917 production and release during aging may contribute to the development of BPH . 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 . 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 . 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 . Dph3 is an electron donor for Dph1-Dph2 in the first step of eukaryotic diphthamide biosynthesis . DB03223 , the target of diphtheria toxin , is a unique posttranslational modification on translation elongation factor 2 ( P13639 ) in archaea and eukaryotes . The biosynthesis of diphthamide was proposed to involve three steps . The first step is the transfer of the 3-amino-3-carboxypropyl group from S-adenosyl-l-methionine ( DB00118 ) to the histidine residue of P13639 , forming a C-C bond . Previous genetic studies showed this step requires four proteins in eukaryotes , Dph1-Dph4 . However , the exact molecular functions for the four proteins are unknown . Previous study showed that Pyrococcus horikoshii Dph2 ( PhDph2 ) , a novel iron-sulfur cluster-containing enzyme , forms a homodimer and is sufficient for the first step of diphthamide biosynthesis in vitro . Here we demonstrate by in vitro reconstitution that yeast Dph1 and Dph2 form a complex ( Dph1-Dph2 ) that is equivalent to the homodimer of PhDph2 and is sufficient to catalyze the first step in vitro in the presence of dithionite as the reductant . We further demonstrate that yeast Dph3 ( also known as KTI11 ) , a Q14406 -type zinc finger protein , can bind iron and in the reduced state can serve as an electron donor to reduce the Fe-S cluster in Dph1-Dph2 . Our study thus firmly establishes the functions for three of the proteins involved in eukaryotic diphthamide biosynthesis . For most radical DB00118 enzymes in bacteria , flavodoxins and flavodoxin reductases are believed to serve as electron donors for the Fe-S clusters . The finding that Dph3 is an electron donor for the Fe-S clusters in Dph1-Dph2 is thus interesting and opens up new avenues of research on electron transfer to Fe-S proteins in eukaryotic cells . Differential expression of P41594 in human lumbosacral motoneurons . Glutamatergic excitotoxicity is one of the main hypotheses to explain motoneuronal degeneration in amyotrophic lateral sclerosis ( P35858 ) . Interestingly , autonomic motoneurons remain almost unaffected , even in late stages of the disease . Since glutamate receptors may mediate neurotoxic as well as neuroprotective effects , different expression patterns may contribute to neuronal vulnerability . We and others have previously described a significantly higher expression of group I metabotropic glutamate receptors ( mGluRs ) in rat autonomic motoneurons compared to somatic motoneurons . Here we show a selective expression of the group I receptor P41594 in human parasympathetic Onuf 's nucleus . These results are in accordance with previous findings in rat and strengthen the hypothesis that mGluR expression may provide a possible clue to the selective vulnerability in P35858 . Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells . Aphidicolin specifically inhibits eukaryotic DNA polymerase alpha , while 2',3'-dideoxythymidine 5'-triphosphate ( d2TTP ) inhibits P06746 and gamma but not alpha . DB00987 5'-triphosphate ( araCTP ) inhibits both DNA polymerase alpha and beta although to a different extent . Here we measured the effects of these inhibitors on repair DNA synthesis of U.V.-irradiated HeLa cells by two different methods . Firstly , aphidicolin , 1-beta-D-arabinofuranosylcytosine ( araC , a precursor of araCTP ) and 2',3'-dideoxythimidine ( d2Thd , a precursor of d2TTP ) were added directly to the culture medium . In this case , aphidicolin and araC strongly inhibited replicative DNA synthesis of HeLa cells , and they also inhibited repair synthesis after U.V.-irradiation but to a much lesser extent . In contrast , high concentrations of d2Thd inhibited repair DNA synthesis to a higher extent than replicative DNA synthesis . Secondly , the active form of inhibitor , d2TTP , was microinjection directly into cytoplasm or nuclei or U.V.-irradiated HeLa cells . Microinjection of d2TTP effectively inhibited repair synthesis . The microinjection of d2TTP , into either cytoplasm or nucleus , strongly inhibited replicative synthesis . These results might indicate that multiple DNA polymerases are involved in repair synthesis as well as in replicative synthesis . The influence of ionotropic and metabotropic glutamate receptor ligands on anxiety-like effect of amphetamine withdrawal in rats . Chronic amphetamine use results in anxiety-like states after drug cessation . The aim of the study was to determine a role of ionotropic and metabotropic glutamate receptor ligands in amphetamine-evoked withdrawal anxiety in the elevated plus-maze test in rats . In our study memantine ( 8 and 12 mg/kg ) , a noncompetitive N-methyl-d-aspartate ( DB01221 ) receptor antagonist did not reduce amphetamine withdrawal anxiety . DB00659 ( DB01221 and metabotropic glutamate 5 receptor ( P41594 ) antagonist ) at the dose 200 and 400mg/kg showed anxiolytic-like effect , thus increasing the percent of time spent in open arms and a number of open arm entries . P41594 selective antagonist , MTEP ( 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine hydrochloride ) and Q14416 /3 agonist , LY354740 ( 1S,2S,5R,6S ) -2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid ) , caused effects similar to acamprosate at doses 1.25-5mg/kg and 2.5-5mg/kg , respectively . None of the glutamate ligands influenced locomotor activity of rats when given to the saline-treated group . Taking into account the positive correlation between amphetamine withdrawal-induced anxiety and relapse to amphetamine taking , our results suggest that modulation of mGluRs may prevent relapse to amphetamine and might pose a new direction in amphetamine abuse therapy . Early decrease of mitochondrial DNA repair enzymes in spinal motor neurons of presymptomatic transgenic mice carrying a mutant P00441 gene . Growing evidence has recently shown that mutant P00441 accumulate in the mitochondria and cause vacuolation in transgenic mice carrying mutant P00441 , an animal model of amyotrophic lateral sclerosis ( P35858 ) . In this study , the expressions of DNA repair enzymes , oxoguanine glycosylase 1 ( ogg1 ) , P06746 ( polbeta ) , and DNA polymerase gamma ( polgamma ) were examined in transgenic mice with an P35858 -linked mutant P00441 gene , a valuable model for human P35858 . In presymptomatic Tg mice , the nuclear form of ogg1 was upregulated , whereas mitochondrial ogg1 remained at the same level . DNA polymerase was selectively downregulated in the mitochondria . This study suggests an impaired protective mechanism against oxidative stress in mitochondria . The expressions of these enzymes are predominant in spinal motor neurons , suggesting a mechanism of selective motor neuron death in this animal model of P35858 . Autoantibodies to tailor-made panels of tumor-associated antigens in breast carcinoma . Autoantibodies ( AAbs ) to tumor-associated antigens ( TAAs ) have been identified in the sera of cancer patients . In a previous review published in this journal , we have focused on recent knowledge related to circulating AAbs to individual TAAs in breast carcinoma . This review will focus on recent knowledge related to AAb assays to tailor-made panels of TAAs in breast carcinoma . So far , AAb assays to the following tailor-made panels of TAAs have been assessed in breast carcinoma : ( 1 ) p53 , c-myc , P04626 , NY-ESO-1 , P51587 , and P15941 , ( 2 ) IMP1 , p62 , Koc , p53 , c-MYC , cyclin B1 , and survivin , ( 3 ) P62937 , P32119 , Q02790 , P10809 , and P15941 , ( 4 ) P15941 , P04626 , p53 , and P18065 , ( 5 ) p53 , P04626 , P18065 , and TOPO2α , ( 6 ) survivin and livin , ( 7 ) Q96DX5 , Q96JX3 , and Q969Z4 , and ( 8 ) p16 , p53 , and c-myc . Assessment of serum AAbs to a tailor-made panel of TAAs provides better sensitivity to diagnosis of breast carcinoma than measuring serum AAbs to a single TAA . Nevertheless , measurement of serum AAbs to a panel of TAAs for screening and early diagnosis of breast carcinoma is still investigational and should be carried out along with traditional diagnostic studies . 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 .

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interacts_with DB01016?

multiple_choice

Differential gene expression in well-regulated and dysregulated pancreatic beta-cell ( MIN6 ) sublines . To identify genes involved in regulated insulin secretion , we have established and characterized two sublines derived from the mouse pancreatic beta-cell line MIN6 , designated B1 and P01024 . They have a similar insulin content , but differ in their secretory properties . B1 responded to glucose in a concentration- and cell confluence-dependent manner , whereas P01024 did not . B1 cells were stimulated by phorbol 12-myristate 13-acetate , leucine , arginine , glibenclamide , isobutylmethylxanthine , and DB00761 , whereas P01024 did not respond ( leucine , arginine , and glibenclamide ) or responded to a lesser extent ( isobutylmethylxanthine , phorbol 12-myristate 13-acetate , and DB00761 ) . Although intracellular Ca(2+) rose in response to glucose in B1 but not P01024 cells , DB00761 increased intracellular Ca(2+) in a similar manner in both sublines . P11166 , P11168 , Kir6.2 , and Q09428 expression was not significantly different between B1 and P01024 cells , whereas P12830 was more abundantly expressed in B1 cells . A more complete list of differentially expressed genes was established by suppression subtractive hybridization and high density ( Affymetrix ) oligonucleotide microarrays . Genes were clustered according to known or putative function . Those involved in metabolism , intracellular signaling , cytoarchitecture , and cell adhesion are of potential interest . These two sublines should be useful for identification of the genes and mechanisms involved in regulated insulin secretion of the pancreatic beta-cell . Cross-talk between the mitogen activated protein kinase and bone morphogenetic protein/hemojuvelin pathways is required for the induction of hepcidin by holotransferrin in primary mouse hepatocytes . BACKGROUND : The circulating hormone hepcidin plays a central role in iron homeostasis . Our goal was to establish an ex vivo iron-sensing model and to characterize the molecular mechanisms linking iron to hepcidin . DESIGN AND METHODS : Murine hepatocytes were isolated by the collagenase method , either from wild type or Q30201 knockout mice , and cultured 42 h without serum before treatments . RESULTS : After 42 h of serum-free culture , hepcidin gene expression was undetectable in the hepatocytes . P81172 gene expression could , however , be re-activated by an additional 24 h of incubation with 10 % serum . Interestingly , addition of 30 microM holotransferrin consistently increased serum-dependent hepcidin levels 3- to 5-fold . The effects of serum and serum+holotransferrin were direct , transcriptional , independent of de novo protein synthesis and required the presence of bone morphogenetic protein . P02787 receptor-2 activation by its ligand holotransferrin led to extracellular signal regulated kinase ( P29323 ) /mitogen activated protein kinase pathway stimulation and the P29323 specific inhibitor U0-126 blunted holotransferrin-mediated induction of hepcidin . P29323 activation by holotransferrin provoked increased levels of phospho- Q15797 /5/8 highlighting cross-talk between the bone morphogenetic protein/hemojuvelin and P27361 /2 pathways . Finally , we demonstrated , using hepatocytes isolated from Hfe(-/-) mice , that Q30201 was not critical for the hepcidin response to holotransferrin but important for basal hepcidin expression . CONCLUSIONS : We demonstrate that hepatocytes are liver iron-sensor cells and that transferrin receptor-2 , by signaling through the P27361 /2 pathway , and bone morphogenetic protein/hemojuvelin , by signaling through the Smad pathways , coordinately regulate the iron-sensing machinery linking holotransferrin to hepcidin . Vitamin D regulates the gut microbiome and protects mice from dextran sodium sulfate-induced colitis . The active form of vitamin D [ DB00136 , 1,25(OH)2D3 ] and the vitamin D receptor ( P11473 ) regulate susceptibility to experimental colitis . The effect of the bacterial microflora on the susceptibility of C57BL/6 mice to dextran sodium sulfate-induced colitis was determined . Mice that can not produce 1,25(OH)2D3 [ Cyp27b1 ( Cyp ) knockout ( KO ) ] , P11473 KO as well as their wild-type littermates were used . Cyp KO and P11473 KO mice had more bacteria from the Bacteroidetes and Proteobacteria phyla and fewer bacteria from the Firmicutes and Deferribacteres phyla in the feces compared with wild-type . In particular , there were more beneficial bacteria , including the Lactobacillaceae and Lachnospiraceae families , in feces from Cyp KO and P11473 KO mice than in feces from wild-type . Helicobacteraceae family member numbers were elevated in Cyp KO compared with wild-type mice . Depletion of the gut bacterial flora using antibiotics protected mice from colitis . 1,25(OH)2D3 treatment ( 1.25 μg/100 g diet ) of Cyp KO mice decreased colitis severity and reduced the numbers of Helicobacteraceae in the feces compared with the numbers in the feces of untreated Cyp KO mice . The mechanisms by which the dysbiosis occurs in P11473 KO and Cyp KO mice included lower expression of P12830 on gut epithelial and immune cells and fewer tolerogenic dendritic cells that resulted in more gut inflammation in P11473 and Cyp KO mice compared with wild-type mice . Increased host inflammation has been shown to provide pathogens with substrates to out-compete more beneficial bacterial species . Our data demonstrate that vitamin D regulates the gut microbiome and that 1,25(OH)2D3 or P11473 deficiency results in dysbiosis , leading to greater susceptibility to injury in the gut . 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 . 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 . Clinical and functional characterization of the Pro1198Leu Q09428 gene mutation associated with permanent neonatal diabetes mellitus . AIMS/INTRODUCTION : The adenosine triphosphate ( DB00171 ) -sensitive potassium ( KATP ) channel is a key component of insulin secretion in pancreatic β-cells . Activating mutations in Q09428 encoding for the sulfonylurea receptor subunit of the KATP channel have been associated with the development of neonatal diabetes mellitus ( NDM ) . The aim was to investigate clinical and functional characterization of the Pro1198Leu Q09428 gene mutation associated with permanent NDM ( PNDM ) . MATERIALS AND METHODS : The coding regions and conserved splice sites of Q14654 , Q09428 and P01308 were screened for mutations in a 12-year-old girl diagnosed with PNDM . The functional property of the mutant channel identified was examined with patch-clamp experiments in COS-1 cells . We also investigated the difference of effectiveness between two groups of oral sulfonylureas in vitro and in the patient . RESULTS : We identified a heterozygous missense mutation ( c.3593 C > T , Pro1198Leu ) in Q09428 . The mutated residue ( P1198 ) is located within a putative binding site of sulfonylureas , such as tolbutamide or gliclazide . In patch-clamp experiments , the mutant channel was less DB00171 sensitive than the wild type . Furthermore , the sensitivity to tolbutamide was also reduced in the mutant channel . In addition to the tolbutamide/gliclazide binding site , glibenclamide is thought to also bind to another site . DB01016 was more effective than other sulfonylureas in vitro and in the patient . The treatment of the patient was finally able to be switched from insulin injection to oral glibenclamide . CONCLUSIONS : We identified the Pro1198Leu Q09428 mutation in a PNDM patient , and clarified the functional and clinical characterization . The present findings provide new information for understanding PNDM . 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 . Monoclonal antibodies against P00533 in non-small cell lung cancer . Blockade of the epidermal growth factor receptor ( P00533 ) by monoclonal antibodies is a strategy to improve outcome in patients with non-small cell lung cancer . Cetuximab , a chimeric anti- P00533 monoclonal antibody , has been studied in combination with different chemotherapy protocols in both phase II and phase III trials in patients with advanced NSCLC . In the phase III FLEX trial , cetuximab added to cisplatin/vinorelbine resulted in an absolute overall survival benefit of 1.2 months compared to the same chemotherapy alone in patients with advanced P00533 -expressing NSCLC . In the second phase III trial , cetuximab added to carboplatin plus paclitaxed failed to improve progression-free survival but suggested a survival benefit similar to that seen in the FLEX trial . However , the benefit in survival reached statistical significance only in the FLEX trial . A meta-analysis that included patients from four randomized trials confirmed the efficacy of cetuximab when added to chemotherapy . Thus addition of cetuximab to platinum-based chemotherapy represents a new treatment option for patients with advanced NSCLC . DB05101 and panitumumab have also been evaluated in phase II trials . DB09559 is currently evaluated in combination with chemotherapy in two randomized phase III trials . 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 . 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 . Induction of apoptosis of Beta cells of the pancreas by advanced glycation end-products , important mediators of chronic complications of diabetes mellitus . We herein report cytotoxicity of advanced glycation end-products ( AGEs ) on pancreatic beta cells . AGEs stimulated reactive oxygen species ( ROS ) generation but did not arrest proliferation of the P01308 -1 cell line . Pancreatic beta cell lines or primary cultured islets possess a receptor for P51606 ( RAGE ) , and its expression increased after P51606 treatment . TUNEL staining and FACS analysis using annexin V/PI antibodies showed that apoptosis increased in P01308 -1 cells or primary cultured islets when incubated with BSA conjugated with glyceraldehyde ( AGE2 ) or glucoaldehyde ( AGE3 ) , compared with those conjugated with glucose ( AGE1 ) . Reaction of P01308 -1 cells to Ki67 , which is a cellular marker for proliferation , was also increased after P51606 treatment . The ability of primary cultured islets to secrete insulin was retained even after P51606 treatment under either low or high glucose conditions . The antiserum against RAGE partially prevented P51606 -induced cellular events . Treatment of beta cells with the antioxidant metallothionein results in a significant reduction in pathologic changes . AGEs might be able to induce apoptosis as well as proliferation of pancreatic beta cell lines or primary cultured islets . Moreover , antibody array showed that Q06609 and P43351 were significantly decreased in AGE2-treated P01308 -1 cells . AGEs might inhibit homologous DNA recombination for repairing DNA of P01308 -1 cells damaged by ROS generation . It might be suggested that treatment of AGEs resulted in ROS production and apoptosis through their receptor on pancreatic beta cells . AGEs might deteriorate function of pancreatic beta cells in patients with long-term hyperglycemia . Structure-function relationships in the beta-cell K( DB00171 ) channel . The DB00171 -sensitive potassium ( K( DB00171 ) ) channel plays a key role in controlling beta-cell membrane potential and insulin secretion . The channels are composed of two subunits , Kir6.2 , which forms the channel pore , and Q09428 , which contains binding sites for nucleotides and sulphonylureas and acts as a channel regulator . Our current studies are aimed at delineating the molecular interactions involved in assembly and ligand binding by K( DB00171 ) channel proteins . We have employed a complementation approach in which Q09428 half-molecules are co-expressed in insect cells using a baculovirus system . Together with data from truncated Q09428 molecules and a fusion protein in which Q09428 is linked to Kir6.2 , we have interpreted our findings in terms of a model for the structure of the K( DB00171 ) channel . The main features of the model are : ( i ) the C-terminal end of Q09428 is close to the N-terminus of Kir6.2 ; ( ii ) the two nucleotide binding domains ( NBDs ) of Q09428 -- NBD1 and NBD2 -- are in proximity ; ( iii ) transmembrane helix 12 of Q09428 is orientated in such a way that it can make contact with Kir6.2 ; ( iv ) formation of the glibenclamide binding site requires that the two cytosolic loops ( CLs ) CL3 and CL8 are located close to each other ; ( v ) there are homomeric interactions between the NBD1 domains of neighbouring subunits . We suggest that binding of glibenclamide leads to conformational changes in CL3 and CL8 leading to rearrangement of transmembrane helices . These effects are transmitted to Kir6.2 to result in channel closure . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . 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 . The glutamate/neutral amino acid transporter family Q99705 : molecular , physiological and pharmacological aspects . The solute carrier family 1 ( Q99705 ) includes five high-affinity glutamate transporters , P43005 , GLT-1 , P43003 , P48664 and O00341 ( P43005 , P43004 , P43003 , P48664 , and O00341 , respectively ) as well as the two neutral amino acid transporters , P43007 and Q15758 ( P43007 and ALC1A5 , respectively ) . Although each of these transporters have similar predicted structures , they exhibit distinct functional properties which are variations of a common transport mechanism . The high-affinity glutamate transporters mediate transport of l- DB00142 , l- DB00128 and d- DB00128 , accompanied by the cotransport of 3 Na(+) and 1 H(+) , and the countertransport of 1 K(+) , whereas ASC transporters mediate Na(+)-dependent exchange of small neutral amino acids such as Ala , DB00133 , DB00151 and DB00156 . The unique coupling of the glutamate transporters allows uphill transport of glutamate into cells against a concentration gradient . This feature plays a crucial role in protecting neurons against glutamate excitotoxicity in the central nervous system . During pathological conditions , such as brain ischemia ( e.g. after a stroke ) , however , glutamate exit can occur due to " reversed glutamate transport " , which is caused by a reversal of the electrochemical gradients of the coupling ions . Selective inhibition of the neuronal glutamate transporter P43005 ( P43005 ) may be of therapeutic interest to block glutamate release from neurons during ischemia . On the other hand , upregulation of the glial glutamate transporter P43004 ( P43004 ) may help protect motor neurons in patients with amyotrophic lateral sclerosis ( P35858 ) , since loss of function of P43004 has been associated with the pathogenesis of certain forms of P35858 . Amsacrine and etoposide hypersensitivity of yeast cells overexpressing DNA topoisomerase II . Increasing the cellular concentration of DNA topoisomerase II in yeast by expressing constitutively a plasmid-borne P11388 gene encoding the enzyme greatly increases the sensitivity of the cells to amsacrine and etoposide ( DB00773 ) . This increased drug sensitivity at a higher intracellular DNA topoisomerase II level is observed in both P43351 + repair-proficient strains and rad52 mutants that are defective in the repair of double-stranded breaks . These results provide strong support of the hypothesis that the cellular target of these drugs is DNA topoisomerase II , and that these drugs kill cells by converting DNA topoisomerase II into a DNA damaging agent . Glycoprotein nonmetastatic melanoma protein B ( Q14956 ) as a novel neuroprotective factor in cerebral ischemia-reperfusion injury . Glycoprotein nonmetastatic melanoma protein B ( Q14956 ) is a type I transmembrane protein reported to have neuroprotective effects in the neurodegenerative disease amyotrophic lateral sclerosis ( P35858 ) . We investigated whether Q14956 is also neuroprotective against brain ischemia-reperfusion injury ( IRI ) . Focal ischemia/reperfusion injury was induced via filament middle cerebral artery occlusion ( MCAO ) for 2h , followed by reperfusion upon withdrawal of the filament . We assessed the neuroprotective effects of Q14956 using transgenic ( Tg ) mice which over expressing Q14956 or recombinant Q14956 which has the sequence of human extracellular Q14956 . The results showed that Q14956 was up-regulated after IRI , and that genomic over-expression of Q14956 significantly ameliorated infarct volume . Next , we investigated the protective mechanisms of Q14956 via Western blotting and immunohistochemistry ( IHC ) . Phosphorylation of extracellular signal-regulated kinase 1 and 2 ( P27361 /2 ) , and protein kinase B ( Akt ) , were increased in the Q14956 Tg group according to Western blotting data . IHC analysis showed that Q14956 was expressed not only in neurons , but also in astrocytes , produced labeling patterns similar to that in human brain ischemia . Furthermore , recombinant Q14956 also decreased infarction volume . These results indicate that Q14956 protected neurons against IRI , and phosphor-Akt and phosphor- P29323 might be a part of the protective mechanisms , and that the neuroprotective effect of Q14956 was seemingly induced by the extracellular sequence of Q14956 . In conclusion , these findings indicate that Q14956 has neuroprotective effects against IRI , via phosphorylation of P27361 /2 and Akt , suggesting that Q14956 may be a therapeutic target for ischemia-reperfusion injuries . DB04958 , a P20273 -targeting recombinant humanized antibody with a different mode of action from rituximab . DB04958 is a humanized anti- P20273 monoclonal antibody currently in clinical trials for treatment of non-Hodgkin lymphoma ( Q9NZ71 ) and certain autoimmune diseases . Here we report the results of investigations of epratuzumab 's mode of action in comparison to and in combination with the anti- P11836 mAb , rituximab . In vitro cell growth inhibition , induction of apoptosis , and the ability of the mAbs to mediate complement-dependent cytotoxicity ( CDC ) and antibody-dependent cellular cytotoxicity ( ADCC ) were evaluated . We also investigated the potential activity of epratuzumab in the regulation of B-cell antigen receptor ( P11274 ) activation . DB04958 and rituximab displayed very distinct modes of action ; epratuzumab acts as an immunomodulatory agent , while rituximab is an acutely cytotoxic therapeutic antibody . DB04958 has distinct effects on cell growth from rituximab . For example , rituximab+anti-human IgG Fcgamma yielded marked inhibition of proliferation in human Q9NZ71 cell lines , while epratuzumab had little or no effect in this assay . However , when cells were immobilized and stimulated with anti-IgM , epratuzumab , but not rituximab , caused a significant antiproliferative effect . Unlike rituximab , no CDC could be detected , and ADCC was modest but significant with epratuzumab . Importantly , combining rituximab and epratuzumab did not decrease rituximab 's ability to induce apoptosis , CDC , and ADCC . In fact , the combination is more effective than rituximab alone in inhibiting proliferation of Daudi Burkitt lymphoma cells in the presence of second antibody , and at least equally effective to rituximab in the absence of crosslinking . These observations suggest that it may be possible to enhance clinical efficacy by combination therapy comprised of anti- P11836 and anti- P20273 mAbs . 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 . c- P05412 promotes P11274 - P00519 -induced lymphoid leukemia by inhibiting methylation of the 5' region of Cdk6 . The transcription factor c- P05412 and its upstream kinase P45983 have been implicated in P11274 - P00519 -induced leukemogenesis . P45983 has been shown to regulate P10415 expression , thereby altering leukemogenesis , but the impact of c- P05412 remained unclear . In this study , we show that P45983 and c- P05412 promote leukemogenesis via separate pathways , because lack of c- P05412 impairs proliferation of p185( P11274 - P00519 )-transformed cells without affecting their viability . The decreased proliferation of c-Jun(Δ/Δ) cells is associated with the loss of cyclin-dependent kinase 6 ( Q00534 ) expression . In c-Jun(Δ/Δ) cells , Q00534 expression becomes down-regulated upon P11274 - P00519 -induced transformation , which correlates with CpG island methylation within the 5' region of Cdk6 . We verified the impact of Cdk6 deficiency using Cdk6(-/-) mice that developed P11274 - P00519 -induced B-lymphoid leukemia with significantly increased latency and an attenuated disease phenotype . In addition , we show that reexpression of Q00534 in P11274 - P00519 -transformed c-Jun(Δ/Δ) cells reconstitutes proliferation and tumor formation in Nu/Nu mice . In summary , our study reveals a novel function for the activating protein 1 ( AP-1 ) transcription factor c- P05412 in leukemogenesis by antagonizing promoter methylation . Moreover , we identify Q00534 as relevant and critical target of AP-1-regulated DNA methylation on P11274 - P00519 -induced transformation , thereby accelerating leukemogenesis .

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interacts_with DB01109?

multiple_choice

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 . DB01269 : the evidence for its use in the treatment of metastatic colorectal cancer . DB01269 is the first fully human monoclonal antibody to Epidermal Growth Factor Receptor ( P00533 ) to enter clinical trials for the treatment of solid tumors . The anti-tumor activity of panitumumab has been tested in vitro and in vivo , and inhibition of tumor growth has been observed in numerous cancer models , particularly lung , kidney and colorectal ( CRC ) . Preclinical and clinical studies have established a role for panitumumab in metastatic colorectal cancer ( mCRC ) refractory to multiple chemotherapeutic regimens . Based on these encouraging findings , panitumumab was approved by the US Food and Drug Administration for the treatment of patients with epidermal growth factor receptor-expressing mCRC refractory to fluoropyrimidine- , oxaliplatin- , and/or irinotecan-containing chemotherapeutic regimens . The improvement in progression free survival ( PFS ) and response rate ( RR ) produced by panitumumab monotherapy was significantly greater in patients with non mutated ( wild-type ) K- DB01367 than in those with mutant K- DB01367 . Therefore implementing routine K- DB01367 screening and limiting the use of P00533 inhibitors to patients with wild-type K- DB01367 appears the better strategy for select only the patients who could benefit from the therapy with panitumumab and also may have the potential for cost savings . The purpose of this review was to evaluate the patient-related , disease-related and economic-related evidence for the use of panitumumab in the treatment of metastatic colorectal cancer in clinical practice . Reduced fibrin deposition and intravascular thrombosis in hDAF transgenic pig hearts perfused with tirofiban . BACKGROUND : Solid organ xenograft rejection is associated with vascular injury resulting at least in part in platelet activation , and rejected xenografts invariably demonstrate intravascular thrombosis and interstitial hemorrhage . Complement activation plays a prominent role in platelet-endothelial interaction . We tested the effects of platelet P08514 /IIIa inhibitor tirofiban during perfusion of hDAF pig hearts . METHODS : Using a working-heart model , nontransgenic and hDAF pig hearts were perfused with tirofiban or human blood only . Myocardial damage was determined by hemodynamic parameters ( cardiac output , stroke work index ) and creatine phosphokinase . Further monitoring included the assessment of complement factors ( P01024 , C4 ) , platelets , fibrinogen , P01008 , and graft histology . RESULTS : Tirofiban increased cardiac output ( CO ) and stroke work index ( SWI ) of nontransgenic pig hearts and improved superior CO and SWI of hDAF pig hearts . Although perfusion time of nontransgenic pig hearts was prolonged by tirofiban ( 196+/-65 min vs. 162+/-122 min ) , a similar effect in hDAF pig hearts ( 218+/-116 min vs. 222+/-30 min ) could not be demonstrated . Tirofiban reduced consumption of P01024 and C4 independently from hDAF . Depletion of fibrinogen was equally diminished by tirofiban and hDAF ; the combination of both agents obtained no further reduction . P01008 consumption was most effectively inhibited by this combination . Intravascular fibrin deposition was reduced by tirofiban and hDAF , but particularly by the combination of the two agents . CONCLUSIONS : Improvement of heart performance and reduction of myocardial damage and intravascular thrombosis confirm a role of the P08514 /IIIa inhibitor tirofiban for the prevention of hDAF pig heart rejection and xenograft function . Molecular dynamics simulation of human neurohypophyseal hormone receptors complexed with oxytocin-modeling of an activated state . The neurohypophyseal hormone oxytocin ( CYIQNCPLG-NH(2) , OT ) is involved in the control of labor , secretion of milk and many social and behavioral functions via interaction with its receptors ( OTR ) located in the uterus , mammary glands and peripheral tissues , respectively . In this paper we propose the interactions responsible for OT binding and selectivity to OTR versus vasopressin ( [ P13726 ,R8]OT , AVP ) receptors : P37288 and P30518 , all three belonging to the Class A G protein-coupled receptors ( GPCRs ) . Three-dimensional models of the activated receptors were constructed using a multiple sequence alignment and the activated rhodopsin-transducin ( MII-Gt ) prototype [ Slusarz and Ciarkowski , 2004 ] as a template . The 1 ns unconstrained molecular dynamics ( MD ) of three pairs of receptor-OT complexes ( two complexes per each receptor ) immersed in the fully hydrated 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine ( POPC ) lipid bilayer was conducted in the AMBER 7.0 force field . The relaxed models of ligand-receptor complexes were used to identify the putative binding sites of OT . The stabilizing interactions with conserved Gln residues in all complexes were identified . The nonconserved hydrophobic residues were proposed as responsible for OTR-OT selectivity and ligand recognition . These results provide guidelines for experimental site-directed mutagenesis and if confirmed , they may be helpful in designing new selective OT analogs with both agonistic or antagonistic properties . P13726 expression in human pterygium . PURPOSE : A pterygium shows tumor-like characteristics , such as proliferation , invasion , and epithelial-mesenchymal transition ( EMT ) . Previous reports suggest that tissue factor ( TF ) expression is closely related to the EMT of tumor cells , and subsequent tumor development . In this study , we analyzed the expression and immunolocalization of TF in pterygial and normal conjunctival tissues of humans . METHODS : Eight pterygia and three normal bulbar conjunctivas , surgically removed , were used in this study . DB03843 -fixed , paraffin-embedded tissues were submitted for immunohistochemical analysis with anti-TF antibody . Double staining immunohistochemistry was performed to assess TF and alpha-smooth muscle actin ( α-SMA ) or epidermal growth factor receptor ( P00533 ) expression in the pterygia . RESULTS : Immunoreactivity for TF was detected in all pterygial tissues examined . TF immunoreactivity was localized in the cytoplasm of basal , suprabasal , and superficial epithelial cells . The number of TF-immunopositive cells in pterygial epithelial cells was significantly higher than in normal conjunctival epithelial cells ( p < 0.001 ) . TF immunoreactivity was detected in α-SMA-positive or -negative pterygial epithelial cells . P00533 immunoreactivity was detected in pterygial epithelium , which was colocalized with TF . CONCLUSIONS : These results suggest that TF plays a potential role in the pathogenesis and development of a pterygium , and that TF expression might be involved through EMT-dependent and -independent pathways . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . 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 . Potential role of activated platelets in homing of human endothelial progenitor cells to subendothelial matrix . Endothelial progenitor cells ( EPCs ) mobilize from the bone marrow in response to tissue injury and participate in vascular repair . However , there is limited data about the homing mechanisms of EPCs to vascular injury sites . Recently animal experiments indicated that platelets play a role in recruitment of EPCs to injury sites . However , data on the possible interaction between platelets and EPCs within the human system are limited . We , therefore , examined in-vitro human platelet-EPC interaction under static and flow conditions . Human EPCs were isolated from donated buffy coats by magnetic microbeads and flow cytometry cell sorting using CD133 and P35968 , respectively , as markers . Platelets were tested in the form of washed platelets , platelet rich plasma or whole blood . EPCs formed heterotypic aggregates with resting platelets under static conditions , an interaction that was greatly enhanced when platelets were activated by collagen , ADP or thrombin-activation peptide . The platelet-EPC interaction was inhibited by antibodies to P16109 or P16109 glycoprotein ligand-1 ( Q14242 ) , but not by antibodies to glycoproteins Ib-IX-V or IIb/IIIa . When perfused over activated platelets under shear stress of 2.5 dyn/cm(2) , EPCs tethered to platelayers and either adhered immediately or rolled a short distance before adhering . In addition , platelets promoted the colonization of adherent EPCs in culture conditions . Consistent with recent animal studies , these findings demonstrate that human EPCs interact in vitro with activated platelets under static and flow conditions , mediated through P16109 - Q14242 interaction . This interaction may be a central mechanism for homing of EPCs to vascular injury sites . Mutation of the histidin residue of the DRH motif in vasopressin V2 receptor expression and function . BACKGROUND AND THE PURPOSE OF THE STUDY : DB00067 type 2 receptor ( P30518 ) , a G protein coupled receptor ( GPCR ) , plays an important role in the regulation of renal antidiuretic function . The highly conserved DRH motif is essential for G protein signaling of P30518 ; however its role especially regarding the histidin residue is not fully understood .. METHODS : Site directed mutagenesis was performed with replacements of the histidin to isoleucine by using nested polymerase chain reaction . ELISA was performed for receptor expression assay and the adenylyl cyclase activity assay was performed for functional characterization of DRI mutation on P30518 signaling . RESULTS AND MAJOR CONCLUSION : The adenylyl cyclase activity assay in COS-7 cells showed no difference in the amount of DB02527 production between the wild type and the mutant V2 receptors . The V2 receptor expression was not changed in the presence of this mutation using ELISA assay . These results suggest that the role of histidin residue is not critical in the V2 receptor function , however further mutagenesis studies are required to define the role of this motif in P30518 function . Developments in epidermal growth factor receptor-targeting therapy for solid tumors : focus on matuzumab ( P50402 72000 ) . Expression of epidermal growth factor and its transmembrane receptor ( P00533 ) stimulates tumor growth . DB05101 is a humanized anti- P00533 monoclonal antibody that blocks P00533 activation and downstream signaling , inhibits tumor growth , and provides a clinical benefit for some patients . The plasma half-life ( 6-10 days ) and pharmacodynamic activity allow flexible dosing on weekly , every-2-week , and every-3-week schedules . DB05101 has shown single-agent antitumor activity in heavily pretreated patients with a variety of tumors , with a favorable safety profile . Skin rash is the most common toxicity , but is severe ( Grade 3 ) in < 1 percent . This article describes preclinical and clinical development of matuzumab . New aspects in the pathogenesis of diabetic atherothrombosis . Diabetes mellitus is increasing worldwide , resulting from the interaction of obesity , inflammation , and hyperglycemia . Activated immunity and cytokine production lead to insulin resistance and other components of the metabolic syndrome , establishing the link between diabetes and atherosclerosis . Hyperglycemia-induced endothelial dysfunction is mediated by increased oxidative stress , a promoter of adventitial inflammation and vasa vasorum neovascularization in experimental models of diabetic atherosclerosis . Recent studies have documented increased inflammation , neovascularization , and intraplaque hemorrhage in human diabetic atherosclerosis . This inflammatory microangiopathic process is independently associated with plaque rupture , leading to coronary thrombosis . P13726 , the most potent trigger of the coagulation cascade , is increased in diabetic patients with poor glycemic control . Circulating tissue factor microparticles are also associated with apoptosis of plaque macrophages , closing the link among inflammation , plaque rupture , and blood thrombogenicity . High-density lipoproteins , responsible for free cholesterol removal , are reduced in patients with insulin resistance and diabetes . High-density lipoprotein therapy leads to a significant decrease in plaque macrophages and increase in smooth-muscle cells . These beneficial effects may be responsible for coronary plaque stabilization in patients treated with recombinant P02647 Milano/phospholipid complex . Finally , peroxisomal proliferator-activated receptors ( PPARs ) are now considered the nuclear transcriptional regulators of atherosclerosis . Three subfamilies , including Q07869 , -delta , and -gamma , have been identified with crucial roles in lipid metabolism , plaque inflammation , expression of adhesion molecules and cytokines , and regulation of matrix metalloproteinases . Multiple experimental studies have documented plaque stabilization with P37231 agonists , a group of medications holding great promise in the treatment of diabetes atherosclerosis . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Concomitant oral and intravenous pharmacokinetics of trametinib , a MEK inhibitor , in subjects with solid tumours . AIMS : The aim of this phase 1 , single centre , open label study in four patients with solid tumours was to determine the absolute bioavailability of a 2 mg oral dose of trametinib . DB08911 is an orally bioavailable , reversible and selective allosteric inhibitor of Q02750 and P36507 activation and kinase activity . METHODS : A microtracer study approach , in which a 5 μg radiolabelled i.v. microdose of trametinib was given concomitantly with an unlabelled 2 mg oral tablet formulation , was used to recover i.v. and oral pharmacokinetic parameters , simultaneously . RESULTS : The least-squares mean ( 90 % confidence interval ) absolute bioavailability of trametinib ( 2 mg tablet ) was 72.3 % ( 50.0 % , 104.6 % ) . Median tmax after oral administration was 1.5 h and the geometric mean terminal half-life was 11 days . The geometric mean clearance and volume of distribution after i.v. administration were 3.21 l h(-1) and 976 l , respectively , resulting in a terminal elimination half-life of 11 days . CONCLUSIONS : DB08911 absolute bioavailability was moderate to high , whereas first pass metabolism was low . P37231 reduces the growth rate of pancreatic cancer cells through the reduction of cyclin D1 . P37231 ( PPARgamma ) forms a heterodimeric DNA-binding complex with the retinoid X receptor ( RXR ) and regulates the transcription of its target genes . Activation of PPARgamma has been shown to induce P55008 arrest and to inhibit cell growth of human pancreatic carcinoma cell lines . The purpose of the present study was to examine the effect of ligand activation of PPARgamma and RXR on cell growth and on the expression of P55008 cyclins in a pancreatic cancer cell line PANC-1 , which expresses PPARgamma at high levels . DB00197 , a specific ligand for PPARgamma , was found to cause a reduction in the growth rate and induced P55008 cell cycle arrest and this effect was additive with that of 9-cis retinoic acid ( 9-cis RA ) , a ligand for RXR . Of the P55008 cyclins tested , troglitazone specifically reduced the expression of cyclin D1 mRNA and the corresponding protein and this effect was also additive with 9-cis RA . These results suggest that the activation of PPARgamma together with RXR may be useful for the suppression of pancreatic cancer cell growth through the reduction in cyclin D1 levels . Inhibition of vascular endothelial growth factor receptor tyrosine kinases impairs adipose tissue development in mouse models of obesity . We have studied the effect of PTK787 ( DB04879 ) , an inhibitor of vascular endothelial growth factor receptor ( VEGFR ) tyrosine kinases , on adipose tissue development . Oral administration of PTK787 for 4 weeks ( 2 mg/g high fat diet , HFD ) to C57Bl/6 mice resulted in a significant reduction in total body weight and of subcutaneous ( SC ) and gonadal ( GON ) adipose tissue mass , as compared to control animals fed HFD only ( all p < 0.0005 ) . In the GON adipose tissue adipocytes were hypertrophic after PTK787 treatment . Blood vessel size and density were not significantly affected by PTK787 treatment . Expression of Flk-1 ( P35968 ) mRNA was significantly reduced in SC and GON adipose tissues of PTK787 treated mice . De novo fat pad formation following injection of preadipocytes in Q9NXR1 mice was significantly ( p < 0.005 ) impaired by PTK787 administration ( 2 mg/g HFD for 4 weeks ) , without associated effect on blood vessel size or density . Thus , in nutritionally induced murine obesity models , oral administration of the VEGFR tyrosine kinases inhibitor PTK787 resulted in reduced adipose tissue development . The differential impact of DB05876 subtypes in human lung fibroblasts on cytokine-induced proliferation and myofibroblast conversion . Lung fibroblast proliferation and differentiation into myofibroblasts are pathological key events during development of lung fibrosis . Cyclic nucleotide signaling is described as a negative modulator of these cellular processes , and cyclic nucleotide degrading type 4 phosphodiesterases ( DB05876 ) are important regulators of these pathways . In this study , we elucidated expression and the role of individual subtypes of DB05876 in primary normal human lung fibroblast ( NHLF ) in controlling cytokines-induced proliferation and conversion to myofibroblasts by short-interfering RNAs ( siRNAs ) induced knockdown . We verified the expression of P27815 , B , and D , while Q08493 was only minor or even not expressed in NHLF . An efficient liposome-mediated transfection method for mRNA silencing and a knockdown of the expressed DB05876 subtypes was achieved in these cells . This knockdown was further validated by DB05876 protein expression analysis and DB05876 activity measurements . Functionally , the knockdown of P27815 and Q07343 inhibited proliferation induced by the cytokine combination of P09038 and IL-1β , whereas knockdown of Q08499 was ineffective . In contrast , TGF-β induced differentiation into myofibroblasts was affected by knockdown of Q07343 and Q08499 , but not by P27815 knockdown . In summary , our data allow to assign different DB05876 subtypes to distinct functions of human lung fibroblasts and highlight the predominant role of Q07343 in controlling pathophysiological processes of human lung fibroblasts . This provides a scientific rationale for focused therapeutic targeting of Q07343 to treat respiratory diseases with fibrotic lesions in the lung . The attenuation of experimental lung metastasis by a bile acid acylated-heparin derivative . The inhibitory efficacies of new bile acid acylated-heparin derivative ( heparin-DOCA ) were evaluated on experimental lung metastasis . We evaluated the effect of heparin-DOCA on intercellular interactions including those between B16F10 and thrombin-activated platelets and P01375 -activated HUVECs , and between B16F10 and immobilized mouse P16109 . In addition , the inhibitory effects of heparin-DOCA on adhesion and invasion of B16F10 to Matrigel were studied . In an animal mouse study , the blood clot formation and the retention of red fluorescence protein ( RFP ) -B16F10 in lungs were assessed after heparin-DOCA and RFP-B16F10 intravenous administration . Furthermore , we investigated the anti-metastatic effect of heparin-DOCA against lung metastasis induced by B16F10 and SCC7 . DB01109 -DOCA inhibited intercellular interactions between B16F10 and activated platelets or activated HUVECs by blocking P- and P16581 -mediated interactions . Moreover , it reduced adhesion and invasion of B16F10 to Q13201 , thereby affecting the reduction of early retention of B16F10 in the lung . DB01109 -DOCA attenuated lung colony formation on the surfaces and in interior of the lung , and attenuated metastasis by B16F10 and SCC7 . These results suggest that heparin-DOCA may have potentials as therapeutic agent that prevents tumor metastasis and progression . A conserved mechanism for gating in an ionotropic glutamate receptor . Ionotropic glutamate receptor ( iGluR ) channels control synaptic activity . The crystallographic structure of P42262 , the prototypical iGluR , reveals a clamshell-like ligand-binding domain ( LBD ) that closes in the presence of glutamate to open a gate on the pore lining α-helix . How LBD closure leads to gate opening remains unclear . Here , we show that bending the pore helix at a highly conserved alanine residue ( Ala-621 ) below the gate is responsible for channel opening . Substituting Ala-621 with the smaller more flexible glycine resulted in a basally active , nondesensitizing channel with ∼39-fold increase in glutamate potency without affecting surface expression or binding . On P42262 (A621G) , the partial agonist kainate showed efficacy similar to a full agonist , and competitive antagonists CNQX and DB03759 acted as a partial agonists . DB00134 -629 in P42262 sits above the gate and is critical in transmitting LBD closure to the gate . Substituting DB00134 -629 with the flexible glycine resulted in reduced channel activity and glutamate potency . The pore regions in potassium channels are structurally similar to iGluRs . Whereas potassium channels typically use glycines as a hinge for gating , iGluRs use the less flexible alanine as a hinge at a similar position to maintain low basal activity allowing for ligand-mediated gating . [ 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 . 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 . A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins . A surface plasmon resonance-based solution affinity assay is described for measuring the K(d) of binding of heparin/heparan sulfate-binding proteins with a variety of ligands . The assay involves the passage of a pre-equilibrated solution of protein and ligand over a sensor chip onto which heparin has been immobilised . DB01109 sensor chips prepared by four different methods , including biotin-streptavidin affinity capture and direct covalent attachment to the chip surface , were successfully used in the assay and gave similar K(d) values . The assay is applicable to a wide variety of heparin/HS-binding proteins of diverse structure and function ( e.g. , P05230 , P09038 , P15692 , P10145 , P80075 , P01008 , P02776 ) and to ligands of varying molecular weight and degree of sulfation ( e.g. , heparin , DB05808 , sucrose octasulfate , naphthalene trisulfonate ) and is thus well suited for the rapid screening of ligands in drug discovery applications . Persistent activation of P27361 /2 by lead acetate increases nucleotide excision repair synthesis and confers anti-cytotoxicity and anti-mutagenicity . Lead , a possible human carcinogen , affects signal transduction pathways in many aspects , yet exhibits low mutagenicity in human cells . In this study , we explore whether signaling pathways including the four MAPKs and AKT affect DNA repair and mutagenicity in the exposure of mammalian cells to lead acetate [ Pb(II) ] . Pb(II) increased the phosphorylated P27361 /2 and phosphorylated AKT but not the phosphorylated Q13164 , phosphorylated p38 and JNK activity in human non-small cell lung adenocarcinoma CL3 cells . The duration of P27361 /2 activation was much longer than AKT activation and these two signals were independently activated by Pb(II) in CL3 cells . Intriguingly , a Q02750 /2 inhibitor PD98059 ( 25-50 micro M ) markedly suppressed P27361 /2 activation and greatly promoted the hprt mutation frequency and cytotoxicity in Pb(II)-treated CL3 cells . Conversely , inhibition of the AKT signal by wortmannin did not exhibit such effects . Inhibition of the persistently activated P27361 /2 in Pb(II)-treated diploid human fibroblasts by PD98059 also markedly increased the mutagenicity and cytotoxicity . The Pb(II)-induced mutagenicity and cytotoxicity were significantly higher in nucleotide excision repair ( P55055 ) -deficient UVL-10 rodent cells than their counterpart P01008 -2 cells ; also , P27361 /2 activation by Pb(II) was observed in P01008 -2 but not UVL-10 cells . Furthermore , cellular P55055 synthesis was enhanced by Pb(II) exposure , which was markedly suppressed by PD98059 . Activation of P27361 /2 by expressing a constitutively active form of Q02750 in CL3 cells also elevated cellular P55055 synthesis . Together , these results indicate that persistent activation of P27361 /2 signaling by Pb(II) enhances cellular P55055 synthesis , thereby conferring anti-cytotoxicity and anti-mutagenicity .

MH_train_1524

MH_train_1524

MH_train_1524

interacts_with DB01285?

multiple_choice

DB01285 -releasing hormone inhibits insulin-like growth factor-I release from primary cultures of rat granulosa cells . DB01285 -releasing hormone ( P06850 ) , a neuropeptide which modulates gonadal function during stress , is expressed by several cell types of the rat ovary and is able to suppress oestrogen release from rat granulosa cells . The mechanism of this effect is , however , not known . Since insulin-like growth factor ( IGF ) -I is produced by rat granulosa cells and exerts a synergistic role with DB00094 on granulosa cell steroidogenesis , we hypothesised that P06850 may suppress oestrogen release from granulosa cells by inhibiting P05019 release and/or stimulating the release of its binding protein ( P17936 ) . To test this hypothesis , granulosa cells were obtained from immature female Sprague-Dawley rats primed with diethylstilboestrol , and hormone concentrations were measured in the conditioned medium by radioimmunoassay . P06850 suppressed oestrogen and P05019 release stimulated by DB00094 used at a concentration of 1 IU/l , whereas it did not have any statistically significant effect on oestrogen and P05019 release in basal conditions or in response to 5 IU/l DB00094 . The suppressive effects of P06850 on oestrogen and P05019 release were antagonised by a selective P06850 receptor antagonist . P06850 had no effects on P17936 release . P06850 did not have any effect on oestrogen release stimulated by increasing concentrations of P05019 and its suppressive effect on DB00094 -stimulated oestrogen release was overcome by the addition of low doses of exogenous P05019 . In conclusion , P06850 suppressed the release of oestrogen and P05019 , but not of P17936 . Thus , the inhibitory effects of P06850 on oestrogen release could be mediated , partly , by a suppression of the autocrine/paracrine action of P05019 . Increased P09601 levels ameliorate fatty liver development through a reduction of heme and recruitment of Q9NSA1 . OBJECTIVE : Obese leptin deficient ( ob/ob ) mice are a model of adiposity that displays increased levels of fat , glucose , and liver lipids . Our hypothesis is that P09601 overexpression ameliorates fatty liver development . METHODS : Obese mice were administered cobalt protoporphyrin ( CoPP ) and stannic mesoporphyrin ( DB04912 ) for 6 weeks . Heme , P09601 , HO activity , PGC1α , Q9NSA1 , glycogen content , and lipogenesis were assessed . RESULTS : CoPP administration increased hepatic P09601 protein levels and HO activity , decreased hepatic heme , body weight gain , glucose levels , and resulted in decreased steatosis . Increased levels of P09601 produced a decrease in lipid droplet size , P49327 ( FAS ) levels involving recruitment of Q9NSA1 , PPARα , and Glut 1 . These beneficial effects were reversed by inhibition of HO activity . CONCLUSION : Increased levels of P09601 and HO activity reduced the levels of obesity by reducing hepatic heme and lipid accumulation . These changes were manifested by decreases in cellular heme , increases in Q9NSA1 , glycogen content , and fatty liver . The beneficial effect of P09601 induction results from an increase in PPARα and Q9NSA1 levels and a decrease in PGC1α , levels they were reversed by DB04912 . Low levels of P09601 and HO activity are responsible for fatty liver . DB06698 ameliorates olanzapine-induced weight gain through modulation of histaminergic , P01303 and AMPK pathways . Olanzapine is widely used to treat schizophrenia and other disorders , but causes adverse obesity and other metabolic side-effects . Both animal and clinical studies have shown that co-treatment with betahistine ( a histaminergic H1 receptor agonist and H3 receptor antagonist ) is effective for ameliorating olanzapine-induced weight gain/obesity . To reveal the mechanisms underlying these effects , this study investigated the effects of co-treatment of olanzapine and betahistine ( O+B ) on expressions of histaminergic H1 receptor ( P35367 ) , AMP-activated protein kinase ( AMPK ) , neuropeptide Y ( P01303 ) , and proopiomelanocortin ( P01189 ) in the hypothalamus associated with reducing olanzapine-induced weight gain . Olanzapine significantly upregulated the mRNA and protein expressions of P35367 , while O+B co-treatment significantly downregulated the P35367 levels , compared to the olanzapine-only treatment group . The P01303 mRNA expression was significantly enhanced by olanzapine , but it was significantly reversed by O+B co-treatment . The hypothalamic P35367 expression was positively correlated with total food intake , and P01303 expression . Olanzapine also increased AMPKα activation measured by the AMPKα phosphorylation ( pAMPKα ) /AMPKα ratio compared with controls , whereas O+B co-treatment decreased the pAMPKα/AMPKα ratio , compared with olanzapine only treatment . The pAMPKα/AMPKα ratio was positively correlated with total food intake and P35367 expression . Although olanzapine administration decreased the P01189 mRNA level , this level was not affected by O+B co-treatment . Therefore , these results suggested that co-treatment with betahistine may reverse olanzapine-induced body weight gain via the P35367 - P01303 and P35367 -pAMPKα pathways . Estimation of skin entrance doses ( SEDs ) for common medical X-ray diagnostic examinations in India and proposed diagnostic reference levels ( DRLs ) . Skin entrance doses ( SEDs ) were estimated by carrying out measurements of air kerma from 101 X-ray machines installed in 45 major and selected hospitals in the country by using a silicon detector-based dose Test-O-Meter . 1209 number of air kerma measurements of diagnostic projections for adults have been analysed for seven types of common diagnostic examinations , viz. chest ( AP , PA , O43561 ) , lumbar spine ( AP , O43561 ) , thoracic spine ( AP , O43561 ) , abdomen ( AP ) , pelvis ( AP ) , hip joints ( AP ) and skull ( PA , O43561 ) for different film-screen combinations . The values of estimated diagnostic reference levels ( DRLs ) ( third quartile values of SEDs ) were compared with guidance levels/DRLs of doses published by the IAEA-BSS-Safety Series No . 115 , 1996 ; Q9Y251 ( O14682 ) ( 2000 and 2005 ) , UK ; CRCPD/CDRH ( USA ) , European Commission and other national values . The values of DRLs obtained in this study are comparable with the values published by the IAEA-BSS-115 ( 1996 ) ; Q9Y251 ( O14682 ) ( 2000 and 2005 ) UK ; EC and CRCPD/CDRH , USA including values obtained in previous studies in India . Genetic demonstration of intestinal Q9UHC9 as a major determinant of hepatic cholesterol and blood atherogenic lipoprotein levels . OBJECTIVE : The correlation between intestinal cholesterol absorption values and plasma low-density lipoprotein-cholesterol ( LDL-C ) levels remains controversial . Niemann-Pick-C1-Like 1 ( Q9UHC9 ) is essential for intestinal cholesterol absorption , and is the target of ezetimibe , a cholesterol absorption inhibitor . However , studies with Q9UHC9 knockout mice or ezetimibe can not definitively clarify this correlation because Q9UHC9 expression is not restricted to intestine in humans and mice . In this study we sought to genetically address this issue . METHODS AND RESULTS : We developed a mouse model that lacks endogenous ( Q9UHC9 ) and P01130 ( P01130 ) ( DKO ) , but transgenically expresses human Q9UHC9 in gastrointestinal tract only ( DKO/ Q9NUQ9 (IntOnly) mice ) . Our novel model eliminated potential effects of non-intestinal Q9UHC9 on cholesterol homeostasis . We found that human Q9UHC9 was localized at the intestinal brush border membrane of DKO/ Q9NUQ9 (IntOnly) mice . DB04540 feeding induced formation of Q9UHC9 -positive vesicles beneath this membrane in an ezetimibe-sensitive manner . Compared to DKO mice , DKO/ Q9NUQ9 (IntOnly) mice showed significant increases in cholesterol absorption and blood/hepatic/biliary cholesterol . Increased blood cholesterol was restricted to very low-density lipoprotein ( VLDL ) and LDL fractions , which was associated with increased secretion and plasma levels of apolipoproteins B100 and B48 . Additionally , DKO/ Q9NUQ9 (IntOnly) mice displayed decreased fecal cholesterol excretion and hepatic/intestinal expression of cholesterologenic genes . DB00973 treatment virtually reversed all of the transgene-related phenotypes in DKO/ Q9NUQ9 (IntOnly) mice . CONCLUSION : Our findings from DKO/ Q9NUQ9 (IntOnly) mice clearly demonstrate that Q9UHC9 -mediated cholesterol absorption is a major determinant of blood levels of apolipoprotein B-containing atherogenic lipoproteins , at least in mice . Current and future pharmacologic options for the management of patients unable to achieve low-density lipoprotein-cholesterol goals with statins . Low-density lipoprotein-cholesterol ( LDL-C ) lowering is the mainstay of the current treatment guidelines in the management of cardiovascular risk . P04035 inhibitors ( statins ) are currently the most effective LDL-C-lowering drugs . However , a substantial number of patients do not reach treatment targets with statins . Therefore , an unmet medical need exists for lipid-lowering drugs with novel mechanisms of action to reach the recommended cholesterol target levels , either by monotherapy or combination therapy . Upregulation of the P01130 with squalene synthase inhibitors has shown promising results in animal studies but the clinical development of the lead compound lapaquistat ( DB05317 ) has recently been discontinued . DB00973 combined with statins allowed significantly more patients to reach their LDL-C targets . Other inhibitors of intestinal cholesterol absorption such as disodium ascorbyl phytostanol phosphate ( DB05449 ) and bile acid transport inhibitors have shown positive results in early development trials , whereas the prospect of acyl coenzyme A : cholesterol acyltransferase inhibition in cardiovascular prevention is dire . Selective inhibition of messenger RNA ( mRNA ) by antisense oligonucleotides is a new approach to modify cholesterol levels . The inhibition of apolipoprotein B mRNA is in advanced development and mipomersen sodium ( ISIS 301012 ) has shown striking results in phase II studies both as monotherapy as well as in combination with statins . Hyperglycemia-Induced Changes in DB08818 Contribute to Impaired Skin Wound Healing in Diabetes : Review and Perspective . Ulcers and chronic wounds are a particularly common problem in diabetics and are associated with hyperglycemia . In this targeted review , we summarize evidence suggesting that defective wound healing in diabetics is causally linked , at least in part , to hyperglycemia-induced changes in the status of hyaluronan ( HA ) that resides in the pericellular coat ( glycocalyx ) of endothelial cells of small cutaneous blood vessels . Potential mechanisms through which exposure to high glucose levels causes a loss of the glycocalyx on the endothelium and accelerates the recruitment of leukocytes , creating a proinflammatory environment , are discussed in detail . Hyperglycemia also affects other cells in the immediate perivascular area , including pericytes and smooth muscle cells , through exposure to increased cytokine levels and through glucose elevations in the interstitial fluid . Possible roles of newly recognized , cross-linked forms of HA , and interactions of a major HA receptor ( P16070 ) with cytokine/growth factor receptors during hyperglycemia , are also discussed . P49327 gene expression in human adipose tissue : association with obesity and type 2 diabetes . AIMS/HYPOTHESIS : Increased expression and activity of the lipogenic pathways in adipose tissue may contribute to the development of obesity . As a central enzyme in lipogenesis , the gene encoding fatty acid synthase ( P49327 ) was identified as a candidate gene for determining body fat . In the present study we tested the hypothesis that increased P49327 expression links metabolic alterations of excess energy intake , including hyperinsulinaemia , dyslipidaemia and altered adipokine profile to increased body fat mass . SUBJECTS AND METHODS : In paired samples of visceral and subcutaneous adipose tissue from 196 participants ( lean or obese ) , we investigated whether P49327 mRNA expression ( assessed by PCR ) in adipose tissue is increased in obesity and related to visceral fat accumulation , measures of insulin sensitivity ( euglycaemic-hyperinsulinaemic clamp ) and glucose metabolism . RESULTS : P49327 mRNA expression was increased by 1.7-fold in visceral vs subcutaneous fat . Visceral adipose tissue P49327 expression was correlated with P49327 protein levels , subcutaneous P49327 expression , visceral fat area , fasting plasma insulin , serum concentrations of P05231 , leptin and retinol-binding protein 4 ( P02753 ) , and inversely with measures of insulin sensitivity , independently of age , sex and BMI . Moreover , we found significant correlations between P49327 expression and markers of renal function , including serum creatinine and urinary albumin excretion . CONCLUSIONS/INTERPRETATION : Increased P49327 gene expression in adipose tissue is linked to visceral fat accumulation , impaired insulin sensitivity , increased circulating fasting insulin , P05231 , leptin and P02753 , suggesting an important role of lipogenic pathways in the causal relationship between consequences of excess energy intake and the development of obesity and type 2 diabetes . 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 . 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 . DB01109 , heparan sulfate and heparanase in cancer : remedy for metastasis ? Malignant tumor cells invade normal tissues in the vicinity of cancer through devastating the extracelluar matrix and blood vessel wall of the tissues . An important step in this process is degradation of heparan sulfate proteoglycan , a carbohydrate-protein complex . P16070 is a major component of the extracellular matrix , and is essential for the self-assembly , insolubility and barrier properties of basement membranes . Q9Y251 is an endoglucuronidase that cleaves heparan sulfate and expression level of this enzyme correlates with metastatic potential of tumor cells . Treatment with heparanase inhibitors markedly reduces the incidence of metastasis in experimental animals . DB01109 , a widely used anticoagulant , is structurally related to heparan sulfate and a natural substrate of heparanase . Long-term treatment of cancer patients having venous thromboembolism with low molecular weight heparin showed improved survival rate . Understanding the functional roles and the corresponding molecular mechanisms of heparin , heparan sulfate and heparanase in cancer development may pave the way for exploring remedies against tumor metastasis . 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 . 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 . 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 . The effect of mevinolin on steroidogenesis in patients with defects in the low density lipoprotein receptor pathway . Adrenal steroidogenesis is dependent upon cholesterol derived from both de novo biosynthesis and uptake of plasma lipoproteins through the low density lipoprotein ( LDL ) receptor pathway . Recent studies have demonstrated that patients homozygous for familial hypercholesterolemia have a mild impairment in cortisol secretion during maximal DB01285 stimulation . DB00227 , a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase , has been used clinically to inhibit de novo cholesterol synthesis in patients with familial hypercholesterolemia . In this study we examined hypothalamic-pituitary-adrenal function in seven patients with defects in the P01130 pathway , both before and during treatment with oral mevinolin ( 20 mg , twice a day ) , to assess whether inhibition of cholesterol synthesis influences steroidogenesis under basal conditions and in response to ovine P06850 and exogenous DB01285 . Two months after initiation of therapy , high density lipoprotein cholesterol levels were significantly elevated , and LDL cholesterol levels were reduced , although not normalized . Basal and ovine P06850 -stimulated adrenocortical function were normal in all patients both before and during therapy . Plateau plasma cortisol concentrations achieved during maximal DB01285 stimulation were lower than those in control subjects in all patients both before and during therapy . All patients , however , had an approximately 3-fold increase over basal values . These results suggest that treatment of patients with defects in the P01130 pathway with mevinolin improves the plasma lipid profile and does not result in adrenal dysfunction or further exacerbation of the mild impairment of adrenal function during maximal DB01285 stimulation . Impact of gene-gender effects of adrenergic polymorphisms on hypothalamic-pituitary-adrenal axis activity in depressed patients . OBJECTIVE : There is overwhelming evidence that activation of the hypothalamic-pituitary-adrenal ( Q9Y251 ) system plays a major role in depression and cardiovascular disease in genetically susceptible individuals . We hypothesized that due to the multiple interactions between the sympathetic and the Q9Y251 systems via adrenoceptors , polymorphisms in these genes could have an impact on Q9Y251 axis activity in major depression . METHODS : Using the dexamethasone/corticotrophin-releasing hormone ( DEX/ P06850 ) test , we investigated the association of alpha(2)-adrenoceptor ( P08913 -1291C --> G ) and the beta(2)-adrenoceptor gene ( P07550 Arg16Gly ) in 189 patients with major depression during the acute state of the disease and after remission . RESULTS : Male P08913 -1291G allele homozygotes showed significant pretreatment Q9Y251 axis hyperactivity , with increased adrenocorticotropin ( DB01285 ; F = 4.9 , d.f. = 2 , p = 0.009 ) and cortisol responses ( F = 6.4 , d.f. = 2 , p = 0.003 ) . In contrast , female P07550 DB00125 / DB00125 homozygotes had increased pretreatment DB01285 ( F = 7.17 , d.f. = 2 , p = 0.001 ) and cortisol ( F = 8.95 , d.f. = 2 , p = 0.000 ) levels . Interestingly , in the respective genotypes , the stress hormones remained elevated in the second DEX/ P06850 test , despite a reduction in depressive symptoms . CONCLUSIONS : This study provides evidence that , depending on gender and polymorphisms , there is continuous Q9Y251 axis overdrive in a proportion of patients irrespective of the status of depression . Considering the importance of stress hormones for cardiovascular disorders , our data might suggest that these patients are at high risk of comorbidity between depression and cardiovascular disorders . Additive role of tiotropium in severe asthmatics and Arg16Gly in P07550 as a potential marker to predict response . BACKGROUND : Recent findings have raised new interests about the use of anticholinergics , especially tiotropium , for the treatment of asthma . This study was performed to determine whether an additional improvement in lung function is obtained when tiotropium is administrated in addition to conventional therapies in severe asthmatics , and to identify factors capable of predicting the response to tiotropium , using a pharmacogenetic approach . METHODS : A total of 138 severe asthmatics on conventional medications and with decreased lung function were randomly recruited . DB01409 18 microg was added once a day and lung functions were measured every 4 weeks . Responders were defined as those with an improvement of > or = 15 % ( or 200 ml ) in the forced expiratory volume in 1 s ( FEV1 ) that was maintained for at least 8 successive weeks . Eleven single nucleotide polymorphisms ( SNPs ) in P11229 -3 ( coding muscarinic receptors one to three ) which were identified by re-sequencing , and Arg16Gly and Gln27Glu in P07550 ( coding beta(2) adrenoreceptor ) were scored in 80 of the 138 asthmatics . RESULTS : Forty-six of the 138 asthmatics ( 33.3 % ) responded to tiotropium treatment . Logistic regression analyses ( controlled for age , gender , and smoking status ) showed that Arg16Gly in P07550 [ P = 0.003 , OR ( 95 % CI ) = 0.21 ( 0.07-0.59 ) in a minor allele-dominant model ] was significantly associated with response to tiotropium . CONCLUSIONS : As many as 30 % of severe asthmatics on conventional medications with reduced lung function were found to respond to adjuvant tiotropium . The presence of Arg16Gly in P07550 may predict response to tiotropium . Zeranol upregulates corticotropin releasing hormone expression in the placental cell line JEG-3 . DB01285 -releasing hormone ( P06850 ) plays a pivotal role in the control of parturition in human . Increased amount of plasma P06850 is associated with pre-mature delivery . Zeranol or α-zearalanol is a mycotoxin produced by fungi in the Fusarium family . Unlike other mycotoxins , exposure to zeranol appears to have minimal health risk . In North America , it is used as a growth-promoting agent in livestock . Because of the health concern of zeranol residue in meat , this practice has not been adopted in Europe . In our study zeranol could induce P06850 protein expression in JEG-3 cells as low as 0.1nM . As electrophoretic mobility shift assay indicated an increase in the CRE binding activity in P06850 promoter , the induction was likely triggered by transcriptional regulation . We further looked into the signal transduction pathway and PKCδ and P27361 /2 were found to be activated . This study showed that zeranol could increase P06850 expression in placental cells , and the findings might be a concern for pregnant women . 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 .

MH_train_1525

MH_train_1525

MH_train_1525

interacts_with DB00215?

multiple_choice

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 . Effects of ghrelin on the proliferation and secretion of splenic T lymphocytes in mice . Ghrelin , a novel gut -- brain peptide predominantly produced by the stomach , displays strong growth hormone ( GH ) -releasing activity mediated by the hypothalamus-pituitary GH secretagogue receptor ( Q92847 ) . Recently , the ghrelin receptor has also been detected in peripheral systems including immune tissues , suggesting that ghrelin may play an important role in the regulation of immune function . In this paper , we assessed the presence and function of the ghrelin receptor in murine splenic T cells . The enriched T cells express the mRNA of ghrelin and ghrelin receptor mRNA , and there is a significantly positive correlation between them . Moreover , we showed that ghrelin dose-dependently inhibits proliferation of splenic T cells when they are costimulated by anti-CD3 . In addition , ghrelin suppressed Th(1) ( P60568 and P01579 ) and Th(2) ( P05112 and P22301 ) cytokines mRNA expression . These results demonstrate the presence of the ghrelin receptor in murine spleen T lymphocytes and a functional role of ghrelin as a modulator of lymphocyte function . This function of ghrelin may have some relevance to the pathophysiology of immunologic alterations related to metabolism . 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 . Bradykinin [ corrected ] B1 receptor antagonism is beneficial in renal ischemia-reperfusion injury . Previously we have demonstrated that bradykinin B1 receptor deficient mice ( B1KO ) were protected against renal ischemia and reperfusion injury ( IRI ) . Here , we aimed to analyze the effect of B1 antagonism on renal IRI and to study whether P46663 knockout or antagonism could modulate the renal expression of pro and anti-inflammatory molecules . To this end , mice were subjected to 45 minutes ischemia and reperfused at 4 , 24 , 48 and 120 hours . Wild-type mice were treated intra-peritoneally with antagonists of either B1 ( R-954 , 200 microg/kg ) or B2 receptor ( HOE140 , 200 microg/kg ) 30 minutes prior to ischemia . Blood samples were collected to ascertain serum creatinine level , and kidneys were harvested for gene transcript analyses by real-time PCR . Herein , P46663 antagonism ( R-954 ) was able to decrease serum creatinine levels , whereas P30411 antagonism had no effect . The protection seen under P46663 deletion or antagonism was associated with an increased expression of GATA-3 , P05112 and P22301 and a decreased T-bet and IL-1beta transcription . Moreover , treatment with R-954 resulted in lower P13500 , and higher P09601 expression . Our results demonstrated that bradykinin P46663 antagonism is beneficial in renal IRI . P62158 -mediated effects of loperamide on chloride transport by brush border membrane vesicles from human ileum . We investigated whether the synthetic opiate loperamide-HCl is able to regulate specific transport systems for sodium and chloride in brush border membrane vesicles ( BBMVs ) from human ileum and whether such activities are mediated by calcium/calmodulin . In BBMVs we studied Na+/H+ antiport , Cl+/OH- antiport , Na+/Cl- cotransport , and the Cl- conductive pathway . Brush border membrane vesicles were incubated with 10 microM loperamide over 4 h at 5 degrees C before the uptake experiments . In ileal BBMVs , loperamide stimulated intravesicular accumulation of Na+ in the presence of Cl- and vice versa . After 1 min of incubation , the stimulatory effect was 35 % +/- 5 % ( p less than 0.005 ) of the control without loperamide . DB00836 also stimulated Cl-/OH- antiport by 30 % +/- 5 % ( p less than 0.005 ) in BBMVs of ileum . In addition , we studied the role of Ca2+/calmodulin in the action of loperamide on chloride transport by human BBMVs . In loperamide-pretreated BBMVs , calmodulin activity was significantly decreased ( 12 +/- 2 vs. 38 +/- 4 pmol/mg protein ) . When loperamide-pretreated vesicles were incubated with 2 microM calcium ( free concentration ) plus 5 microM calmodulin for 1 h at 5 degrees C , complete inhibition of the stimulatory effect of loperamide on Cl-/OH- antiport and Na+/Cl- cotransport was observed . Increasing the Ca2+/calmodulin activity of loperamide-pretreated BBMVs with 2 microM calcium plus 5 microM calmodulin led to a significant inhibition of Cl-/OH- antiport and Na+/Cl- cotransport by 40 % +/- 5 % ( p less than 0.005 ) . In vivo effects of a combined P28222 receptor/ P31645 antagonist in experimental pulmonary hypertension . AIMS : A mechanism for co-operation between the serotonin ( 5-hydroxytryptamine , 5-HT ) transporter and P28222 receptor in mediating pulmonary artery vasoconstriction and proliferation of pulmonary artery smooth muscle cells has been demonstrated in vitro . Here we determine , for the first time , the in vivo effects of a combined P28222 receptor/serotonin transporter antagonist ( LY393558 ) with respect to the development of pulmonary arterial hypertension ( PAH ) and its in vitro effects in human pulmonary artery smooth muscle cells ( hPASMCs ) derived from idiopathic PAH ( IPAH ) patients . METHODS AND RESULTS : We determined the effects of LY393558 as well as a selective serotonin transporter inhibitor , citalopram , on right ventricular pressure , right ventricular hypertrophy , and pulmonary vascular remodelling in wildtype mice and mice over-expressing serotonin transporter ( P31645 + mice ) before and after hypoxic exposure . We also compared their effectiveness at reversing PAH in P31645 + mice and hypoxic mice . Further , we examined the proliferative response to serotonin in IPAH hPASMCs . We also clarified the pharmacology of serotonin-induced vasoconstriction and P28222 receptor/serotonin transporter interactions in mouse isolated pulmonary artery . DB00215 had a moderate effect at preventing and reversing experimental PAH in vivo whereas LY393558 was more effective . LY393558 was more effective than citalopram at reversing serotonin-induced proliferation in IPAH hPASMCs . There is synergy between P28222 receptor and serotonin transporter inhibitors against serotonin-induced vasoconstriction in mouse pulmonary arteries . CONCLUSION : P28222 receptor and serotonin transporter inhibition are effective at preventing and reversing experimental PAH and serotonin-induced proliferation of PASMCs derived from IPAH patients . Targeting both the serotonin transporter and P28222 receptor may be a novel therapeutic approach to PAH . An antibody-lectin sandwich assay for the determination of Q8WXI7 antigen in ovarian cancer patients . A two-step forward sandwich assay was developed for the determination of the ovarian tumour associated glycoconjugate antigen Q8WXI7 with anti- Q8WXI7 Monoclonal antibody Q8TCY5 .1 on the solid phase and 125I-labelled wheat germ lectin as tracer in the solution phase . This Mab-lectin heterosandwich assay was optimized and the clinical utility was evaluated in sera from healthy volunteers and ovarian cancer patients . A correlation was established between Mab-lectin assay and the dual monoclonal antibody sandwich assay , TRUQUANTOV2 RIA , that uses the same MAb Q8TCY5 .1 on the solid phase and a second 125I-labelled DB04964 MAb in the solution phase . A potentially improved clinical utility is suggested for the Mab-lectin assay . The unique format seems to identify novel isoforms of Q8WXI7 with different carbohydrate side chains that would otherwise be undetectable in the MAb-MAb sandwich assay wherein the paratopes are likely directed to protein determinants . 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 . Expression of human all-trans-retinoic acid receptor beta and its ligand-binding domain in Escherichia coli . DB00755 , one of the hormonally active derivatives of vitamin A , occurs physiologically in plasma at a concentration below 10 nmol/l . The methods currently used for its quantification are based on HPLC , need about 1 ml of serum , are relatively laborious and thus not well suited for mass analysis . The affinity and specificity of retinoic acid receptors for all-trans-retinoic acid encouraged us to express both the entire human retinoic acid receptor beta ( P10826 ) and two versions of its retinoic acid-binding domain in Escherichia coli in the hope that these recombinant proteins might be used as binders in a ligand-binding assay for all-trans-retinoic acid . The recombinant receptors , the whole receptor [ P10826 -( Q93033 -Q448) ] , corresponding to domains A-F , and the ligand-binding domain [ P10826 -(E149-Q448) ] , corresponding to domains D-F , were expressed in the vector pET 3d/BL21 ( DE3 ) as inclusion bodies , solubilized with guanidinium chloride , renatured and purified by ion-exchange chromatography . P10826 -(P193-Q448) , corresponding to domains E-F , was expressed in the vector pET 3d/BL21(DE3)pLysS , and purified by reversed-phase chromatography . Under non-denaturing conditions , the expressed whole receptor [ P10826 -( Q93033 -Q448) ] and the D-F construct ( P10826 -(E149-Q448) ] behaved chromatographically as monomeric proteins whereas the E-F construct [ P10826 -(P193-Q448) ] had a strong tendency to aggregate . P10826 -( Q93033 -Q448) and P10826 -(E149-Q448) had similar Kd values for all-trans-retinoic acid ( 1.4 and 0.6 nmol/l respectively ) whereas P10826 -(P193-Q448) bound all-trans-retinoic acid less avidly ( Kd 9.6 nmol/l ) . DB00523 bound to P10826 -(E149-Q448) and P10826 -( Q93033 -Q448) as avidly as all-trans-retinoic acid . Competition experiments showed weak or no binding of 4-oxo-all-trans-retinoic acid , 4-oxo-13-cis-retinoic acid , 13-cis-retinoic acid , acitretin and retinol by P10826 -(E149-Q448) . 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 . Interactions between tumor cells and tumor infiltrating lymphocytes in human ovarian carcinoma . The aim of our study was to investigate the lymphocytic infiltration rate of ovarian tumors and the possible use of tumor-infiltrating lymphocytes ( Q15399 ) as a therapeutic tool in gynecologic oncology . Twelve tumors were treated with digesting enzymes in order to isolate tumor-infiltrating lymphocytes as well as tumor cells , Q15399 were expanded by culture in the presence of human interleukin-2 ( P60568 ) . Freshly prepared tumor cells were allowed to grow in culture medium for several days before the first investigations were performed . Q15399 could only be isolated from 50 % of the investigated tumors . In contrast to this the isolated tumor cells could be largely expanded in 73 % of the cases . The expression of the Q8WXI7 antigen in the culture supernatants served as control and could still be evaluated up to three months after isolation . In parallel the antigen expression on the cellular surface was estimated by immunocytochemistry . Evaluating the phenotypes of Q15399 showed predominantly CD3+ , their expansion rate was only poor . Tumor cells were isolated and expanded in order to test the tumor-directed cytotoxic efficacy of Q15399 and for further use in transplantation to nude mice . Immunomodulatory drugs inhibit expression of cyclooxygenase-2 from P01375 , IL-1beta , and LPS-stimulated human PBMC in a partially P22301 -dependent manner . Immunomodulatory drugs ( IMiDs ) are potent inhibitors of P01375 and IL-1beta and elevators of P22301 production in LPS-stimulated human PBMC . They are currently in clinical trials for various diseases , including multiple myeloma , myelodysplastic syndrome , and melanoma . In the present study , we have investigated the effects of thalidomide , DB00480 and CC-4047 on the expression of P35354 by stimulated PBMC . Our results show that thalidomide and IMiDs inhibited the expression of P35354 but not the P23219 protein in LPS- P01375 and IL-1beta stimulated PBMC and shortened the half-life of P35354 mRNA in a dose-dependent manner . They also inhibited the synthesis of prostaglandin E2 from LPS-stimulated PBMC . While anti- P01375 or IL-1beta neutralizing antibodies had no effect on P35354 expression , anti- P22301 neutralizing antibody elevated the expression of P35354 mRNA , and protein from treated PBMC . These data suggest that the anti-inflammatory and anti-tumor effects of IMiDs may be due in part to elevation of P22301 production and its subsequent inhibition of P35354 expression . Glucocorticoids regulate calcineurin-dependent trans-activating pathways for interleukin-2 gene transcription in human T lymphocytes . Glucocorticoids ( GC ) inhibit P60568 gene transcription by interfering with the binding of the nuclear factor activator protein-1 on the P60568 promoter . Calcineurin , a Ca2+/calmodulin-dependent protein phosphatase , is an essential component of the T cell antigen receptor signal transduction pathway leading to P60568 gene transcription . Therefore , we have asked whether this phosphatase may also be regulated by GC . Jurkat T cells were cotransfected with plasmids containing the intact P60568 promoter or its NF-AT and P14859 motifs , and a deletion mutant ( delta P62158 -AI ) of calcineurin known to have Ca(2+)-independent constitutive phosphatase activity . Cotransfection of P60568 promoter with delta P62158 -AI allowed the activation of P60568 promoter in the presence of phorbol ester alone . Under these conditions dexamethasone ( DB00514 ; 10(-6) M ) inhibited P60568 promoter activation by 50-60 % . The inhibitory effect of DB00514 was specific , as demonstrated by experiments using an unrelated promoter ( simian virus 40 ) and estradiol . Furthermore , it was completely reversed in the presence of excess amounts of the glucocorticoid antagonist RU 486 , which suggests that it is mediated through the glucocorticoid receptor . Overexpression of calcineurin via delta P62158 -AI in Jurkat cells decreased their apparent sensitivity to DB00514 ( approximately 5-fold increase in IC50 ) . Similar results were obtained with the NF-AT and P14859 constructs , which are also known to be activated by calcineurin . Thus , in addition to their known inhibitory effects on activator protein-1 , GC also inhibit calcineurin-dependent pathways for T cell activation . CpG DNA activation and plasma-cell differentiation of P26842 - naive human B cells . Unmethylated CpG DNA activation of naive P26842 - B cells has been reported to require B-cell-receptor ( P11274 ) cross-linking . We describe a culture system using CpG DNA with sequential steps for T-cell-independent activation of naive P15391 + P26842 - human peripheral blood B cells that induces efficient CD138+ plasma-cell differentiation . P26842 + and P26842 - B cells were cultured in a 3-step system : ( 1 ) days 0 to 4 : CpG , P60568 /10/15 ; ( 2 ) days 4 to 7 : P60568 /6/10/15 and anti- P29965 ; ( 3 ) days 7 to 10 : P05231 /15 , IFN-alpha , hepatocyte growth factor , and hyaluronic acid . Both P26842 + and P26842 - B cells up-regulated intracytoplasmic TLR-9 following CpG DNA activation . P26842 - B-cell activation required cell-cell contact . Both naive and memory B cells progressed to a plasma-cell phenotype : CD19lowCD20lowCD27+ P28907 +HLA-DRlow . Seventy percent of the P26842 -- derived CD138+ cells demonstrated productive V chain rearrangements without somatic mutations , confirming their origin from naive precursors . Plasma cells derived from P26842 + B cells were primarily IgG+ , while those from P26842 - B cells were IgM+ . Our results indicate that under certain conditions , naive B cells increase TLR-9 expression and proliferate to CpG DNA stimulation without P11274 signaling . In addition to its immunologic significance , this system should be a valuable method to interrogate the antigenic specificity of naive B cells . 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 . 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 . Erosive arthritis and hepatic granuloma formation induced by peptidoglycan polysaccharide in rats is aggravated by prasugrel treatment . Administration of the thienopyridine Q9H244 receptor antagonist , clopidogrel , increased the erosive arthritis induced by peptidoglycan polysaccharide ( PG-PS ) in rats or by injection of the arthritogenic K/BxN serum in mice . To determine if the detrimental effects are caused exclusively by clopidogrel , we evaluated prasugrel , a third-generation thienopyridine pro-drug , that contrary to clopidogrel is mostly metabolized into its active metabolite in the intestine . Prasugrel effects were examined on the PG-PS-induced arthritis rat model . Erosive arthritis was induced in Lewis rats followed by treatment with prasugrel for 21 days . Prasugrel treated arthritic animals showed a significant increase in the inflammatory response , compared with untreated arthritic rats , in terms of augmented macroscopic joint diameter associated with significant signs of inflammation , histomorphometric measurements of the hind joints and elevated platelet number . Moreover , fibrosis at the pannus , assessed by immunofluorescence of connective tissue growth factor , was increased in arthritic rats treated with prasugrel . In addition to the arthritic manifestations , hepatomegaly , liver granulomas and giant cell formation were observed after PG-PS induction and even more after prasugrel exposure . Cytokine plasma levels of P01584 , P05231 , MIP1 alpha , MCP1 , Q16552 and RANTES were increased in arthritis-induced animals . P22301 plasma levels were significantly decreased in animals treated with prasugrel . Overall , prasugrel enhances inflammation in joints and liver of this animal model . Since prasugrel metabolites inhibit neutrophil function ex-vivo and the effects of both clopidogrel and prasugrel metabolites on platelets are identical , we conclude that the thienopyridines metabolites might exert non-platelet effects on other immune cells to aggravate inflammation . [ Is DB08901 ( DB08901 ) the next treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia ? ] . Distinct clinicopathologic acute lymphoblastic leukemia ( ALL ) entities have been identified , resulting in the adoption of risk-oriented treatment approaches . In Philadelphia chromosome-positive ( Ph(+) ) ALL , the optimal treatment requires the addition of P11274 - P00519 tyrosine kinase inhibitors , as imatinib . However , the outcome remains poor in absence of allogeneic stem cell transplantation , and novel agents are desperately required . Resistance attributable to kinase domain mutations can lead to relapse despite the development of second-generation compounds , including dasatinib and nilotinib . Despite these therapeutic options , the cross-resistant P11274 - P00519 ( T315I ) mutation remains a major clinical challenge . The first evaluations of DB08901 present this drug as a potent multi-targeted kinase inhibitor active against T315I and all other P11274 - P00519 mutants . DB08901 could be the next treatment of choice in hematological malignancies with Philadelphia-positive chromosome , particularly Ph(+) ALL known for its frequent occurrence of T315I mutation . The high glucose-induced stimulation of P46663 and P30411 expression via CB(1)R activation is involved in rat podocyte apoptosis . AIMS : We examined renal kallikrein-kinin system ( KKS ) apoptosis and its related signaling pathway in rat podocytes . In addition , we studied the relationship of cannabinoid receptor 1 ( CB(1)R ) with high glucose and BK receptors . MAIN METHODS : Cell viability was determined by an MTT assay and apoptosis by DNA fragmentation assay , while gene expression was investigated by RT-PCR . Protein expression was analyzed by Western blot analysis . A chemical inhibitor or siRNA transfection was used to inhibit P46663 , P30411 , and CB(1)R signaling . KEY FINDINGS : High glucose ( 25 mM ) treatment decreased cell viability and increased DNA fragmentation . High glucose-induced DNA fragmentation and PARP and caspase-3 activations were blocked by both [ des- DB00125 (10) ] - DB06196 ( a P46663 antagonist ) and DB06196 ( a P30411 antagonist ) . High glucose also increased Akt phosphorylation , ER stress-related protein expression , and NF-κB/I-κB phosphorylation in podocytes , which was blocked by both [ des- DB00125 (10) ] -HOE 140 and DB06196 . In addition , P46663 and P30411 siRNA transfections prevented high glucose-induced Akt and NF-κB activations in rat podocytes . Moreover , AM251 ( a CB(1)R antagonist ) treatment and CB(1)R siRNA transfection blocked the high glucose-induced stimulation of BK receptor expression , Akt activation , and NF-κB activation . SIGNIFICANCE : Our study suggests that hyperglycemia induces apoptosis via the stimulation of P46663 and P30411 expression through CB(1)R activation in rat podocytes in vitro , which is associated with the development of diabetic nephropathy . 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 .

MH_train_1526

MH_train_1526

MH_train_1526

interacts_with DB00734?

multiple_choice

The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis . BACKGROUND : Published data indicate that thyroid stimulating hormone receptor ( P16473 ) activities are associated with osteoporosis in some patients . AIM : This study aimed to elucidate whether a given polymorphism of the P16473 gene is associated with osteoporosis . MATERIALS AND METHODS : One hundred and fifty subjects with osteoporosis were recruited in this study . The diagnosis of osteoporosis was performed with quantitative ultrasound system . The P16473 gene polymorphism was examined by polymerase chain reaction-restriction fragment length polymorphism . RESULTS : The results showed a nucleotide substitution in the first position of codon 36 of the P16473 gene . The nucleotide substitution was from G to C , leading to a (36)D → (36)H change ( D36H ) in the predicted amino acid sequence of the receptor . The change did not show significance between healthy subjects and patients with osteoporosis ( P > 0.05 ) . On the other hand , we identified another single nucleotide polymorphism that is a C-to-G substitution at codon 727 ( GAC to GAG ) ; its frequency was significantly higher in patients with osteoporosis than that in healthy subjects . Using logistic regression analysis , significant correlation was revealed between the genotype D727E and the serum levels of DB00024 , or the quantitative ultrasound value of the calcaneal bone . CONCLUSIONS : The present study suggests that the genotype D727E of the P16473 , but not the genotype D36H , may be a genetic risk factor for osteoporosis . Stress-induced alterations in P08908 receptor transcriptional modulators O75398 and Q6P1N0 . The effect of stress on the mRNA and protein level of the P08908 receptor and two of its key transcriptional modulators , O75398 and Q6P1N0 , was examined in the prefrontal cortex ( P27918 ) and hippocampus ( Hp ) using rodent models : olfactory bulbectomy ( OB ) and prenatal stress ( PS ) in male and female rats ; chronic mild stress in male rats ( CMS ) and pregnancy stress . In P27918 , CMS induced the most widespread changes , with significant reduction in both mRNA and protein levels of O75398 , P08908 receptor and in Q6P1N0 mRNA ; while in Hp P08908 receptor and Q6P1N0 protein levels were also decreased . In male , but not female OB rats P27918 Q6P1N0 and P08908 receptor protein levels were reduced , while in Hp P08908 receptor , Q6P1N0 and O75398 mRNA 's but not protein were reduced . In PS rats P27918 P08908 receptor protein was reduced more in females than males ; while in Hp Q6P1N0 protein was increased in females . In pregnancy stress , P27918 O75398 , Q6P1N0 and P08908 protein receptor levels were reduced , and in HP P08908 receptor protein levels were also reduced ; in HP only O75398 and Q6P1N0 mRNA levels were reduced . Overall , CMS and stress during pregnancy produced the most salient changes in P08908 receptor and transcription factor expression , suggesting a primary role for altered transcription factor expression in chronic regulation of P08908 receptor expression . By contrast , OB ( in males ) and PS ( in females ) produced gender-specific reductions in P27918 P08908 receptor protein levels , suggesting a role for post-transcriptional regulation . These and previous data suggest that chronic stress might be a key regulator of O75398 / Q6P1N0 gene expression . Switching brain serotonin with oxytocin . Serotonin ( 5-HT ) and oxytocin ( P01178 ) are two neuromodulators involved in human affect and sociality and in disorders like depression and autism . We asked whether these chemical messengers interact in the regulation of emotion-based behavior by administering P01178 or placebo to 24 healthy subjects and mapping cerebral 5-HT system by using 2'-methoxyphenyl-(N-2'-pyridinyl)-p-[(18)F]fluoro-benzamidoethylpiperazine ( [(18)F]MPPF ) , an antagonist of P08908 receptors . P01178 increased [(18)F]MPPF nondisplaceable binding potential ( BPND ) in the dorsal raphe nucleus ( DRN ) , the core area of 5-HT synthesis , and in the amygdala/hippocampal complex , insula , and orbitofrontal cortex . Importantly , the amygdala appears central in the regulation of 5-HT by P01178 : [(18)F]MPPF BPND changes in the DRN correlated with changes in right amygdala , which were in turn correlated with changes in hippocampus , insula , subgenual , and orbitofrontal cortex , a circuit implicated in the control of stress , mood , and social behaviors . P01178 administration is known to inhibit amygdala activity and results in a decrease of anxiety , whereas high amygdala activity and 5-HT dysregulation have been associated with increased anxiety . The present study reveals a previously unidentified form of interaction between these two systems in the human brain , i.e. , the role of P01178 in the inhibitory regulation of 5-HT signaling , which could lead to novel therapeutic strategies for mental disorders . Levels of prolactin in relation to coagulation factors and risk of venous thrombosis . Results of a large population-based case-control study ( MEGA-study ) . The pituitary hormone prolactin is thought to influence coagulation . We aimed to study the relation between prolactin levels , coagulation factors and risk of venous thrombosis ( VT ) . We used data from a large population based case-control study into aetiology of first VT ( MEGA-study ) . P01236 levels were determined in 2,068 patients with VT and 2,785 age- and sex matched control subjects . The relation between levels of coagulation factors and prolactin was studied among the controls . In addition , odds ratios ( OR ) and 95 % confidence intervals ( 95 % CI ) were calculated for the risk of VT for different cut-off points of prolactin levels based on percentiles determined in the controls . Restricted analysis was performed among cases in whom blood was sampled within six months after VT . We found a rise in factor VIII and P04275 with increasing levels of prolactin in the controls . An increased risk of VT was observed when blood was sampled within six months after thrombosis ( OR 2.9 , 95 % CI 1.1-8.1 ) for prolactin levels above the 99th percentile ( 42.6 μg/l ) relative to levels between the 20th to 80th percentile . When blood was sampled more than six months after VT no clear association could be observed ( OR 1.3 , 95 % CI 0.7-2.3 ) . In conclusion , we found a modest association between prolactin and symptomatic venous thromboembolism , particularly when blood was sampled close to the event . This may be explained by a causal relation or by prolactin being a marker of stress due to the thrombotic event . The interaction between the oxytocin and pain modulation in headache patients . DB00107 ( P01178 ) , a nonapeptide hormone of posterior pituitary , reaches the central nervous system from systemic blood circulation with a difficulty because of the blood-brain barrier ( BBB ) . The interest has been expressed in the use of the nasal route for delivery of P01178 to the brain directly , exploiting the olfactory pathway . Our previous study has demonstrated that P01178 in the central nervous system rather than the blood circulation plays an important role in rat pain modulation . The communication tried to investigate the interaction between the P01178 and pain modulation in Chinese patients with headache to understand the P01178 effect on human pain modulation . The results showed that ( 1 ) intranasal P01178 could relieve the human headache in a dose-dependent manner ; ( 2 ) P01178 concentration in both plasma and cerebrospinal fluid ( P04141 ) increased significantly in headache patients in relation with the pain level ; and ( 3 ) there was a positive relationship between plasma and P04141 P01178 concentration in headache patients . The data suggested that intranasal P01178 , which was delivered to the central nervous system through olfactory region , could treat human headache and P01178 might be a potential drug of headache relief by intranasal administration . 2(3H)-benzoxazolone and bioisosters as " privileged scaffold " in the design of pharmacological probes . The 2(3H)-benzoxazolone heterocycle and its bioisosteric surrogates ( such as 2(3H)-benzothiazolinone , benzoxazinone , etc. ) have received considerable attention from the medicinal chemists owing to their capacity to mimic a phenol or a catechol moiety in a metabolically stable template . These heterocycles and pyrocatechol have indeed similar pKa 's , electronic charge distribution , and chemical reactivity . Therapeutic applications of this template are very broad , and range from analgesic anti-inflammatory compounds ( including P37231 antagonists ) to antipsychotic and neuroprotective anticonvulsant compounds . High affinity ligands have been obtained also for dopaminergic ( D2 and D4 ) , serotoninergic ( P08908 and P28223 ) , sigma-1 and sigma-2 receptors . Owing to the high number of positive hits encountered with this heterocycle and its congeners , 2(3H)-benzoxazolone template certainly deserves the title of " privileged scaffold " in medicinal chemistry . Inactivation of medium-chain acyl- DB01992 dehydrogenase by oct-4-en-2-ynoyl- DB01992 . Mitochondrial medium-chain acyl- DB01992 dehydrogenase is a key enzyme for the beta-oxidation of fatty acids , which catalyzes the DB03147 -dependent oxidation of a variety of acyl- DB01992 substrates to the corresponding trans-2-enoyl- DB01992 thioesters . Q01860 -en-2-ynoyl- DB01992 was identified as a new irreversible inhibitor of acyl- DB01992 dehydrogenase , and kinetic parameters K(I) and k(inact) were determined to be 11 microM and 0.025 min(-1) , respectively . Triple bond between P06681 and P01024 of the inhibitor was identified as the functional group responsible for enzyme inactivation , and Michael addition is proposed as the mechanism for this inactivation , which is a new pathway for inactivation of P11310 by inhibitors . The inhibitor may become a lead for further development for treating non-insulin-dependent diabetes mellitus . Regulation of the expression of key genes involved in HDL metabolism by unsaturated fatty acids . The cardioprotective effects of HDL have been largely attributed to their role in the reverse cholesterol transport pathway , whose efficiency is affected by many proteins involved in the formation and remodelling of HDL . The aim of the present study was to determine the effects , and possible mechanisms of action , of unsaturated fatty acids on the expression of genes involved in HDL metabolism in HepG2 cells . The mRNA concentration of target genes was assessed by real-time PCR . Protein concentrations were determined by Western blot or immunoassays . Q07869 and liver X receptor ( LXR ) activities were assessed in transfection experiments . Compared with the SFA palmitic acid ( PA ) , the PUFA arachidonic acid ( AA ) , EPA and DB01708 significantly decreased apoA-I , DB00171 -binding cassette A1 ( O95477 ) , lecithin-cholesterol acyltransferase ( P04180 ) and phospholipid transfer protein mRNA levels . EPA and DB01708 significantly lowered the protein concentration of apoA-I and P04180 in the media , as well as the cellular O95477 protein content . In addition , DB01708 repressed the apoA-I promoter activity . AA lowered only the protein concentration of P04180 in the media . The activity of Q07869 was increased by DB01708 , while the activity of LXR was lowered by both DB01708 and AA , relative to PA . The regulation of these transcription factors by PUFA may explain some of the PUFA effects on gene expression . The observed n-3 PUFA-mediated changes in gene expression are predicted to reduce the rate of HDL particle formation and maturation . 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 . The von Willebrand inhibitor DB05202 reduces cerebral embolization after carotid endarterectomy : a randomized trial . BACKGROUND AND PURPOSE : Inhibition of P04275 offers a novel approach to prevention of stroke and myocardial ischemia but has not yet been demonstrated to show efficacy on clinically relevant end points . DB05202 is an aptamer that inhibits the prothrombotic function of P04275 by binding to the A1 domain of P04275 and thereby blocking its interaction with glycoprotein . Phase 1 studies suggest it inhibits platelet aggregation with less increase in bleeding than conventional antiplatelet agents . The effect of ARC 1779 on cerebral emboli immediately after carotid endarterectomy was investigated in a randomized clinical trial . METHODS : Patients undergoing carotid endarterectomy were randomized double-blind to DB05202 or placebo administered intravenously . Transcranial Doppler recording , to detect cerebral embolic signals , was performed in the first 3 hours postoperatively . The primary end point was time to first embolic signals . RESULTS : Thirty-six patients were recruited , 18 in each arm . The Kaplan-Meier median time to first embolic signals was 83.6 minutes for DB05202 compared with 5.5 minutes for placebo . Using Cox proportional hazards embolic signals occurred statistically significantly later on DB05202 ( P=0.007 ) . Reduced embolic signals counts were correlated with inhibition of P04275 activity ( P=0.03 ) . Increased perioperative bleeding and anemia were seen with DB05202 . CONCLUSIONS : P04275 inhibition reduces thromboembolism in humans . It may play a role in treatment of stroke and myocardial ischemia . The extent to which bleeding complications occur in nonoperated patients needs to be assessed in further studies . Clinical Trial Registration- URL : http://clinicaltrials.gov . Unique identifier : NCT00742612 . 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 . Technique appraisement of comparative proteomics and screening of differentiation-related protein in gastric carcinoma . BACKGROUND/AIMS : Different differentiations of cancer have resulted in its unique biological characteristics . We screen and appraise differentially expressed proteins in different differentiated gastric adenocarcinoma with comparative proteomics technology in order to find regulatory factors of tumor differentiation related and finally reach the purpose of tumor differentiation reversal . METHODOLOGY : With two-dimensional fluorescence difference gel electrophoresis ( 2-D DIGE ) and liquid chromatography in conjunction with tandem mass spectrometry ( LC–MS/MS ) , the differentially expressed proteins from 8 patients with different differentiated gastric adenocarcinoma were identified and some factors identified were verified with application of QPCR and Western blot techniques . RESULTS : Significant differences in 35 protein spots were found and 48 kinds of proteins were identified . Other than structural proteins and non-specific protein , six possible proteins associated with tumor differentiation were determined - the serine protease inhibitor B1 ( serine protease inhibitor , clade B , member 1 , P30740 ) , calcium-phospholipid binding protein III ( annexin A3 ) , transcription factor Nm23-H1 , adenine phosphoribosyl-transferase enzyme P07741 ( DB00173 Phosphoribosyltransferase in APO and AMP ) , glutathione S-transferase P1-1 ( Q86UG4 -π-1 ) , antimicrobial peptides P81605 -lL . The identified P30740 , annexin A3 , Nm23-H1 and P07741 were verified , confirming the expression of these factors was in line with proteomics identification . CONCLUSIONS : Protein expression in different differentiated gastric adenocarcinoma was varied . Underexpression of PPARgamma is associated with aneuploidy and lower differentiation of thyroid tumours of follicular origin . P37231 ( PPARgamma ) gene is a nuclear receptor that is involved in thyroid tumourigenesis . Recently , our group has shown that follicular carcinomas underexpressing PPARgamma protein are more prone to develop distant metastases , to invade locally , to present poorly differentiated areas and to persist after surgery . Aneuploidy is also observed in some thyroid tumours , particularly in the more advanced cases . The aim of the present study was to investigate the association of PPARgamma expression with the degree of differentiation and ploidy status of benign and malignant thyroid neoplasias . DNA cytometric studies , ploidy and S-phase fraction ( O76054 ) determination , and quantitative RT-PCR analysis of molecular markers specific for thyroid follicular cells , namely Tg ( thyroglobulin ) , P16473 ( DB00024 receptor ) and Q92911 ( Na+/I- symporter ) were compared between thyroid lesions with positive or negative PPARgamma protein expression . We observed that PPARgamma-negative tissues expressed lower levels of Tg mRNA [ 4.66 x 10(6) a.u. ( arbitrary units ) +/- 1.49 x 10(6) ] , and were more frequently aneuploid ( 36 % ) , and presented higher O76054 ( 3.1 % +/-0.4 ) than PPARgamma-positive samples ( Tg mRNA = 2.54 x 10(7) a.u. +/- 9.72 x 10(6) , P=0.0006 ; aneuploidy=8 % , P=0.0031 ; O76054 =2.2 % +/-0.2 , P=0.0430 ) . A similar trend was also observed for P16473 and Q92911 mRNA , although not reaching statistical significance . This study showed that underexpression of PPARgamma is associated with poor tumour differentiation , aneuploidy and higher cell proliferative activity . Therapies designed to modulate expression of PPARgamma may have an impact on the growth of thyroid neoplasias . 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 . Anti-inflammatory activity of topical THC in DNFB-mediated mouse allergic contact dermatitis independent of P21554 and CB2 receptors . BACKGROUND : ∆(9) - DB00470 ( THC ) , the active constituent of Cannabis sativa , exerts its biological effects in part through the G-protein-coupled P21554 and CB2 receptors , which were initially discovered in brain and spleen tissue , respectively . However , THC also has P21554 /2 receptor-independent effects . Because of its immune-inhibitory potential , THC and related cannabinoids are being considered for the treatment of inflammatory skin diseases . Here we investigated the mechanism of the anti-inflammatory activity of THC and the role of P21554 and CB2 receptors . METHODS : We evaluated the impact of topically applied THC on DNFB-mediated allergic contact dermatitis in wild-type and P21554 /2 receptor-deficient mice . We performed immunohistochemical analyses for infiltrating immune cells and studied the influence of THC on the interaction between T cells , keratinocytes and myeloid immune cells in vitro . RESULTS : Topical THC application effectively decreased contact allergic ear swelling and myeloid immune cell infiltration not only in wild-type but also in P21554 /2 receptor-deficient mice . We found that THC ( 1 ) inhibited the production of IFNγ by T cells , ( 2 ) decreased the production of P13500 and of IFNγ-induced P80075 and CXL10 by epidermal keratinocytes and ( 3 ) thereby limited the recruitment of myeloid immune cells in vitro in a P21554 /2 receptor-independent manner . CONCLUSIONS : Topically applied THC can effectively attenuate contact allergic inflammation by decreasing keratinocyte-derived pro-inflammatory mediators that orchestrate myeloid immune cell infiltration independent of P21554 /2 receptors . This has important implications for the future development of strategies to harness cannabinoids for the treatment of inflammatory skin diseases . LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Molecular sampling of the allosteric binding pocket of the DB00024 receptor provides discriminative pharmacophores for antagonist and agonists . The P16473 ( thyrotropin receptor ) is activated endogenously by the large hormone thyrotropin and activated pathologically by auto-antibodies . Both activate and bind at the extracellular domain . Recently , SMLs ( small-molecule ligands ) have been identified , which bind in an allosteric binding pocket within the transmembrane domain . Modelling driven site-directed mutagenesis of amino acids lining this pocket led to the delineation of activation and inactivation sensitive residues . Modified residues showing CAMs ( constitutively activating mutations ) indicate signalling-sensitive positions and mark potential trigger points for agonists . Silencing mutations lead to an impairment of basal activity and mark contact points for antagonists . Mapping these residues on to a structural model of P16473 indicates locations where an SML may switch the receptor to an inactive or active conformation . In the present article , we report the effects of SMLs on these signalling-sensitive amino acids at the P16473 . Surprisingly , the antagonistic effect of SML compound 52 was reversed to an agonistic effect , when tested at the P62158 Y667A . Switching agonism to antagonism and the reverse by changing either SMLs or residues covering the binding pocket provides detailed knowledge about discriminative pharmacophores . It prepares the basis for rational optimization of new high-affinity antagonists to interfere with the pathogenic activation of the P16473 . P08908 receptor activation : short-term effects on the mRNA expression of the P08908 receptor and galanin in the raphe nuclei . Systemic administration of the P08908 receptor agonist 8-OH-2-(di-n-propylamino)-tetralin ( 8-OH-DPAT ; 0.3 mg/kg , s.c. ) was used to explore the effects of activation of P08908 receptors on expression of mRNA coding for P08908 receptor , tryptophan hydroxylase ( P17752 ) and galanin in the ascending raphe nuclei . 8-OH-DPAT increased the hybridization signal of the P08908 receptor by 105 % in the dorsal raphe nucleus ( P33681 ) 30 min after the injection . No effects were seen at the later time points ( 2-8 h ) . In the median raphe nucleus ( B8 ) and the B9 cell group in the medial lemniscus , 8-OH-DPAT induced a marked decrease in labeling 30 min after injection . At 8 h following 8-OH-DPAT injection , the effect had shifted to an increase in P08908 receptor labeling by 68 % in the B8 area . Importantly 8-OH-DPAT had no significant effects on the expression of mRNA coding for P17752 and galanin . The results suggest an important and differential mechanism for the regulation of P08908 receptor mRNA levels in the dorsal and median raphe nuclei . This regulation may be of importance for the differential control of the activity of the ascending 5-HT neurons , and hence for mood regulation . The results also indicate a dissociation between the effects mediated by P08908 receptor functions and those regulating the coexisting peptide galanin in the dorsal raphe . DB06681 and sirolimus prolong nonhuman primate islet allograft survival : adverse consequences of concomitant alefacept therapy . Calcineurin inhibitors ( CNI ) and steroids are known to promote insulin resistance , and their avoidance after islet transplantation is preferred from a metabolic standpoint . DB06681 , a P33681 -specific mediator of costimulation blockade ( CoB ) , is clinically indicated as a CNI alternative in renal transplantation , and we have endeavored to develop a clinically translatable , belatacept-based regimen that could obviate the need for both CNIs and steroids . Based on the known synergy between CoB and P42345 inhibition , we studied rhesus monkeys undergoing MHC-mismatched islet allotransplants treated with belatacept and the P42345 inhibitor , sirolimus . To extend prior work on CoB-resistant rejection , some animals also received P06729 blockade with alefacept ( P19256 -Ig ) . Nine rhesus macaques were rendered diabetic with streptozotocin and underwent islet allotransplantation . All received belatacept and sirolimus ; six also received alefacept . DB06681 and sirolimus significantly prolonged rejection-free graft survival ( median 225 days compared to 8 days in controls receiving basiliximab and sirolimus ; p = 0.022 ) . The addition of alefacept provided no additional survival benefit , but was associated with Cytomegalovirus reactivation in four of six animals . No recipients produced donor-specific alloantibodies . The combination of belatacept and sirolimus successfully prevents islet allograft survival in rhesus monkeys , but induction with alefacept provides no survival benefit and increases the risk of viral reactivation . 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 . P14416 occupancy by risperidone : implications for the timing and magnitude of clinical response . The objective of the study is to investigate whether dopamine D2 receptor occupancy by risperidone and plasma levels over time can account for therapeutic efficacy and the latency period to response . Thirty-eight examinations with (123)I-IBZM single photon emission computed tomography were performed on 22 patients with schizophrenia , at diagnosis , 48 h after starting risperidone treatment and at a stable dose . DB00734 plasma levels were determined and psychopathologic evaluations ( Brief Psychiatric Rating Scale , Positive and Negative Syndrome Scale ) were carried out . No differences in the striatal/occipital ( S/O ) ratio or plasma levels were found between examinations at the 48-h time point and when a stable dose level had been established , so these parameters could not account for the latency period required for clinical response . D2 receptor occupancy at 48 h correlated positively with clinical improvement after 2 weeks of treatment . Therefore , if these results are confirmed , D2 receptor occupancy at the beginning of treatment with risperidone may be a predictor of subsequent clinical response . 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 . Identification of promising antigenic components in latent fingermark residues . An analysis of latent fingermark residues by Sodium-Dodecyl-Sulfate PolyAcrylamide Gel Electrophoresis ( SDS-PAGE ) followed by silver staining allowed the detection of different proteins , from which two major bands , corresponding to proteins of 56 and 64 kDa molecular weight , could be identified . Two other bands , corresponding to proteins of 52 and 48 kDa were also visualizable along with some other weaker bands of lower molecular weights . In order to identify these proteins , three antibodies directed against human proteins were tested on western blots of fingermarks residues : anti-keratin 1 and 10 ( P04264 /10 ) , anti-cathepsin-D ( Cat.D ) and anti-dermcidin ( Derm. ) . The corresponding antigens are known to be present in the stratum corneum of desquamating stratified epithelium ( P04264 /10 , Cat.D ) and/or in eccrine sweat ( Cat.D , Derm. ) . The two major bands were identified as consistent with keratin 1 and 10 . The pro-form and the active form of the cathepsin-D have also been identified from two other bands . P81605 could not be detected in the western blot . In addition , these antibodies have been tested on latent fingermarks left on polyvinylidene fluoride ( PVDF ) membrane , as well as on whitened and non-whitened paper . The detection of fingermarks was successful with all three antibodies . T cell costimulation : a rational target in the therapeutic armamentarium for autoimmune diseases and transplantation . T cells are central mediators of adaptive immunity . As such , they are involved in both normal immune responses ( e.g. , rejection of a transplanted organ ) and abnormal ones ( e.g. , rheumatoid arthritis ) . T cells require both antigen-specific and costimulatory signals for their full activation . Advances in protein engineering and an increased understanding of the immune response have culminated in the evolution and creation of protein therapeutics that target specific costimulatory molecules . The selective costimulation modulator abatacept ( CTLA-4Ig ) binds to P33681 and P42081 , blocking interaction with P10747 , and is approved for the treatment of moderate to severe rheumatoid arthritis . DB06681 , currently enrolling phase III trials in renal transplantation , was rationally designed from abatacept to bind with more avidity to P42081 , providing the more potent immunosuppressive properties required for immunosuppression in transplantation . This review describes the relevant immunology and summarizes recent clinical findings on these two molecules . Although both inhibit the P10747 costimulatory pathway , they are tailored for specific disease states -- abatacept for autoimmune diseases and belatacept for transplantation . Effects of short- and long-term risperidone treatment on prolactin levels in children with autism . BACKGROUND : The effects of short- and long-term risperidone treatment on serum prolactin were assessed in children and adolescents with autism . METHODS : Patients with autism ( N = 101 , 5-17 years of age ) were randomized to an 8-week trial of risperidone or placebo and 63 then took part in a 4-month open-label follow-up phase . Serum samples were obtained at Baseline and Week-8 ( N = 78 ) , and at 6-month ( N = 43 ) and 22-month ( N = 30 ) follow-up . Serum prolactin was determined by immunoradiometric assay ; dopamine type-2 receptor ( P14416 ) polymorphisms were genotyped . RESULTS : Baseline prolactin levels were similar in the risperidone ( N = 42 ) and placebo ( N = 36 ) groups ( 9.3 +/- 7.5 and 9.3 +/- 7.6 ng/ml , respectively ) . After 8 weeks of risperidone , prolactin increased to 39.0 +/- 19.2 ng/ml , compared with 10.1 +/- 8.8 ng/ml for placebo ( p < .0001 ) . P01236 levels were also significantly increased at 6 months ( 32.4 +/- 17.8 ng/ml ; N = 43 , p < .0001 ) and at 22 months ( N = 30 , 25.3 +/- 15.6 ng/ml , p < .0001 ) . P01236 levels were not associated with adverse effects and P14416 alleles ( Taq1A , -141C Ins/Del , C957T ) did not significantly influence baseline levels or risperidone-induced increases in prolactin . CONCLUSIONS : DB00734 treatment was associated with two- to four-fold mean increases in serum prolactin in children with autism . Although risperidone-induced increases tended to diminish with time , further research on the consequences of long-term prolactin elevations in children and adolescents is needed . 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 . 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 . Perimenopausal regulation of steroidogenesis in the nonhuman primate . Human aging is characterized by a marked decrease in circulating levels of dehydroepiandrosterone ( DB01708 ) and DB01708 -sulfate ( DHEAS ) , hormonal changes associated with cognitive decline . Despite beneficial effects of DB01708 supplementation in rodents , studies in elderly humans have generally failed to show cognitive improvement after treatment . In the present study we evaluate the effects of age and estradiol supplementation on expression of genes involved in the de novo synthesis of DB01708 and its conversion to estradiol in the rhesus macaque hippocampus . Using reverse transcription polymerase chain reaction ( RT-PCR ) we demonstrate the expression of genes associated with this synthesis in several areas of the rhesus brain . Furthermore , real-time PCR reveals an age-related attenuation of hippocampal expression level of the genes P05093 , STS , and 3BHSD1/2 . Additionally , short-term administration of estradiol is associated with decreased expression of P05093 , STS , O00204 , and AROMATASE , consistent with a downregulation not only of estrogen synthesis from circulating DB01708 , but also of de novo DB01708 synthesis within the hippocampus . These findings suggest a decline in neurosteroidogenesis may account for the inefficacy of DB01708 supplementation in elderly humans , and that central steroidogenesis may be a function of circulating hormones and menopausal status . Arsenic decreases antinociceptive activity of paracetamol : possible involvement of serotonergic and endocannabinoid receptors . We assessed whether repeated arsenic exposure can decrease paracetamol-mediated antinociception by modulating serotonergic and endocannabinoid pathways . Rats were preexposed to elemental arsenic ( 4ppm ) as sodium arsenite through drinking water for 28 days . Next day paracetamol 's ( 400mg/kg , oral ) antinociceptive activity was assessed through formalin-induced nociception . Serotonin content and gene expression of P08908 , 5- Q13049 and P21554 receptors were evaluated in brainstem and frontal cortex . Arsenic decreased paracetamol-mediated analgesia . DB00316 , but not arsenic , increased serotonin content in these regions . Arsenic attenuated paracetamol-mediated increase in serotonin level . DB00316 did not alter P08908 expression , but caused down-regulation of 5- Q13049 and up-regulation of P21554 receptors . Arsenic down-regulated these receptors . However , paracetamol-mediated down-regulation of 5- Q13049 was more pronounced . Arsenic did not modify paracetamol 's effect on P08908 expression , but reduced paracetamol-mediated down-regulation of 5- Q13049 and reversed up-regulation of P21554 receptors . Results suggest arsenic reduced paracetamol-induced analgesia possibly by interfering with pronociceptive 5- Q13049 and antinociceptive P21554 receptors . 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 . 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 . Administration of adenosine diphosphate-ribosyl transferase antagonist allows in vivo control of anti-dinitrophenyl response . DB03073 ( 3MB ) is one of a series of chemical inhibitors of the nuclear enzyme adenosine diphosphate ( ADP ) -ribosyl transferase ( P09874 ) , which has been shown to inhibit cell differentiation in vitro , but has no effect on differentiation independent proliferation . Treatment of mice with an optimal concentration of 3MB ( 20 mg/kg body weight ) at or 1 day after dinitrophenyl-keyhole limpet haemocyanin ( DNP-KLH ) immunisation reduced anti-DNP plaque-forming cell ( P27918 ) numbers to less than 10 % of those of control animals . The period for maximum P27918 suppression showed a narrow time window relative to immunisation , suggesting that in vivo , as in vitro , 3 MB was acting only on those lymphocytes differentiating in response to antigen . Experimental findings showed that it was possible to select for P27918 derived from different populations of DNP-responsive lymphocytes by adjusting the time of 3MB treatment relative to immunisation . When 3MB was used with antigen priming , the residual P27918 showed a lower average affinity than P27918 in mice treated with 3MB 3 days after priming , suggesting a differential selection of those lymphocytes responding either ' early ' or ' late ' in the primary immune response . 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 . A bridging interaction allows calmodulin to activate NO synthase through a bi-modal mechanism . P62158 ( P62158 ) activates the nitric-oxide synthases ( NOS ) by a mechanism that is not completely understood . A recent crystal structure showed that bound P62158 engages in a bridging interaction with the NOS Q68DA7 subdomain . We investigated its importance in neuronal NOS ( P29475 ) by mutating the two residues that primarily create the bridging interaction ( DB00125 (752) in the Q68DA7 subdomain and DB00142 (47) in P62158 ) . Mutations designed to completely destroy the bridging interaction prevented bound P62158 from increasing electron flux through the Q68DA7 subdomain and diminished the Q68DA7 -to-heme electron transfer by 90 % , whereas mutations that partly preserve the interaction had intermediate effects . The bridging interaction appeared to control Q68DA7 subdomain interactions with both its electron donor ( NADPH- DB03147 subdomain ) and electron acceptor ( heme domain ) partner subdomains in P29475 . We conclude that the DB00125 (752)- DB00142 (47) bridging interaction is the main feature that enables P62158 to activate P29475 . The mechanism is bi-modal and links a single structural aspect of P62158 binding to specific changes in P29475 protein conformational and electron transfer properties that are essential for catalysis . DB00470 -induced neurotoxicity depends on P21554 receptor-mediated c-Jun N-terminal kinase activation in cultured cortical neurons . Delta9- DB00470 ( THC ) , the main psychoactive ingredient of marijuana , induces apoptosis in cultured cortical neurons . THC exerts its apoptotic effects in cortical neurons by binding to the P21554 cannabinoid receptor . The P21554 receptor has been shown to couple to the stress-activated protein kinase , c-Jun N-terminal kinase ( JNK ) . However , the involvement of specific JNK isoforms in the neurotoxic properties of THC remains to be established . The present study involved treatment of rat cultured cortical neurons with THC ( 0.005-50 microM ) , and combinations of THC with the P21554 receptor antagonist , AM 251 ( 10 microM ) and pertussis toxin ( PTX ; 200 ng ml-1 ) . Antisense oligonucleotides ( AS ) were used to deplete neurons of P45983 and P45984 in order to elucidate their respective roles in THC signalling . Here we report that THC induces the activation of JNK via the P21554 receptor and its associated G-protein , Gi/o . Treatment of cultured cortical neurons with THC resulted in a differential timeframe of activation of the P45983 and P45984 isoforms . Use of specific P45983 and P45984 AS identified activation of caspase-3 and DNA fragmentation as downstream consequences of P45983 and P45984 activation . The results from this study demonstrate that activation of the P21554 receptor induces JNK and caspase-3 activation , an increase in Bax expression and DNA fragmentation . The data demonstrate that the activation of both P45983 and P45984 isoforms is central to the THC-induced activation of the apoptotic pathway in cortical neurons . Simultaneous oxytocin and arg-vasopressin measurements in microdialysates using capillary liquid chromatography-mass spectrometry . DB00107 ( P01178 ) and arg-vasopressin ( AVP ) are nonapeptides with many important functions both peripherally and centrally . Intracerebral microdialysis has helped characterize their importance in regulating complex social and emotional processes . Radioiummunoassay is the most commonly used analytical method used for P01178 and AVP measurements in microdialysates . These measurements have several well-known issues including single peptide per assay limit , possible cross-reactivity between structurally related peptides , and laborious sample preparation with radioactive materials . Here we demonstrate the use of capillary LC-MS(3) for measuring P01178 and AVP simultaneously in dialysates at a 10 min sampling frequency . Microdialysate samples required no preparation and instrumentation was commercially available . Microdialysis probes made with polyacrylonitrile membranes were suitable for high level recovery of the peptides in vitro and in vivo . Responses were linear from 1 to 100 pM . Matrix effect was assessed by standard addition experiments and by comparing signal intensities of P01178 and AVP standards made in aCSF or dialysate . It was determined that the online washing step used on this setup was adequate for removing contaminants which interfere with electrospray ionization efficiency . In vivo , both peptides were stimulated by high K(+) ( 75 mM ) aCSF perfusion in the paraventricular nucleus ( PVN ) . Also , a systemic injection of high Na(+) ( 2M ) caused a rapid and transient increase in PVN P01178 while AVP increased only after 1.5h . Our findings suggest that capillary LC-MS(3) is a straightforward method for monitoring P01178 and AVP simultaneously from complex samples such as dialysates . 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 .

MH_train_1527

MH_train_1527

MH_train_1527

interacts_with DB06684?

multiple_choice

Clinical endocannabinoid deficiency ( CECD ) : can this concept explain therapeutic benefits of cannabis in migraine , fibromyalgia , irritable bowel syndrome and other treatment-resistant conditions ? OBJECTIVES : This study examines the concept of clinical endocannabinoid deficiency ( CECD ) , and the prospect that it could underlie the pathophysiology of migraine , fibromyalgia , irritable bowel syndrome , and other functional conditions alleviated by clinical cannabis . METHODS : Available literature was reviewed , and literature searches pursued via the National Library of Medicine database and other resources . RESULTS : Migraine has numerous relationships to endocannabinoid function . Anandamide ( AEA ) potentiates P08908 and inhibits 5- Q13049 receptors supporting therapeutic efficacy in acute and preventive migraine treatment . Cannabinoids also demonstrate dopamine-blocking and anti-inflammatory effects . AEA is tonically active in the periaqueductal gray matter , a migraine generator . THC modulates glutamatergic neurotransmission via DB01221 receptors . Fibromyalgia is now conceived as a central sensitization state with secondary hyperalgesia . Cannabinoids have similarly demonstrated the ability to block spinal , peripheral and gastrointestinal mechanisms that promote pain in headache , fibromyalgia , IBS and related disorders . The past and potential clinical utility of cannabis-based medicines in their treatment is discussed , as are further suggestions for experimental investigation of CECD via P04141 examination and neuro-imaging . CONCLUSION : Migraine , fibromyalgia , IBS and related conditions display common clinical , biochemical and pathophysiological patterns that suggest an underlying clinical endocannabinoid deficiency that may be suitably treated with cannabinoid medicines . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . Neurotensin interacts with dopaminergic neurons in rat brain . Neurotensin ( NT ) injected intracerebroventricularly in rat increases dopamine ( DA ) turnover in the corpus striatum and nucleus accumbens . Significant increases in 3,4-dihydroxyphenylacetic acid ( DOPAC ) levels occurred within 15 minutes after injection with peak levels at 60 minutes . The effect on NT on DOPAC and homovanillic acid ( HVA ) accumulation was dose-dependent at 3-100 micrograms . NT , like haloperidol , stimulated 3,4-dihydroxyphenylalanine ( DOPA ) accumulation in striatal neurons , in the presence of P20711 inhibitor , after injection of DB04699 ( Q9BVC4 ) . NT had a similar stimulatory effect on DOPA levels in the accumbens while haloperidol ( 0.25 mg.kg-1 ) had no significant effect in this brain region . NT did not block the inhibitory effect of apomorphine on DOPA accumulation in both the striatum and accumbens , while haloperidol inhibited apomorphine effect in both regions . NT also failed to displace 3H-spiperone from DA receptors and the presence of NT in the binding assay did not alter the ability of DA to displace 3H-spiperone in either brain region . These experiments demonstrate that NT increases DA turnover in both the nigrostriatal and mesolimbic pathways . 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 . Dual coupling and down regulation of human DB00094 receptor in CHO cells . The DB00094 receptor is a member of the family of G protein-coupled receptors that activate adenylyl cyclase . The binding of agonist to cell surface receptors leads to a reduction in the intensity of the response to continuous stimulation , a process that is usually referred to as desensitization . Although the exact mechanism is not fully understood , the molecular cloning of the DB00094 receptor has made it possible to study desensitization in transfected cell lines . In this experiment DB00094 -induced desensitization was studied using Chinese hamster ovary cells expressing a functional human DB00094 receptor ( CHO- P23945 cells ) . Stimulation of the CHO- P23945 cells with 10 ng/ml human DB00094 resulted in a decreased sensitivity to a second DB00094 stimulation . This decrease in DB00094 -induced DB02527 production was observed within 2 h , and exposure of cells to DB00094 for 20 h led to a 70-80 % inhibition of DB02527 formation . Moreover , the desensitization effect observed in CHO cells was mimicked by forskolin and , therefore , was mediated by DB02527 . Incubation of cells with 125I- DB00094 showed an efficient internalization of the ligand in the CHO- P23945 cells . The CHO- P23945 cells rapidly internalized approximately 30 % of the receptor-associated 125I- DB00094 by 2 h and 50 % by 4 h . The responsiveness of individual CHO- P23945 cells to DB00094 was studied and administration of human DB00094 ( 30 ng/ml ) induced a rapid rise in cytosolic calcium , reaching a peak at 6 sec . The data that human DB00094 can increase intracellular calcium in cells transfected with the DB00094 receptor cDNA reveal the possibility for the human DB00094 receptor to couple to both adenylyl cyclase and phospholipase C cascades . 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 . 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 . [(11)C]5-HTP and microPET are not suitable for pharmacodynamic studies in the rodent brain . The PET tracer [(11)C]5-hydroxytryptophan ( [(11)C]5-HTP ) , which is converted to [(11)C]5-hydroxytryptamine ( [(11)C]5-HT ) by aromatic amino acid decarboxylase ( P20711 ) , is thought to measure 5-HT synthesis rates . But can we measure these synthesis rates by kinetic modeling of [(11)C]5-HTP in rat ? Male rats were scanned with [(11)C]5-HTP ( 60 minutes ) after different treatments . Scans included arterial blood sampling and metabolite analysis . 5-HT synthesis rates were calculated by a two-tissue compartment model ( 2TCM ) with irreversible tracer trapping or Patlak analysis . DB00190 ( inhibitor peripheral P20711 ) dose-dependently increased [(11)C]5-HTP brain uptake , but did not influence 2TCM parameters . Therefore , 10 mg/kg carbidopa was applied in all subsequent study groups . These groups included treatment with NSD 1015 ( general P20711 inhibitor ) or p-chlorophenylalanine ( PCPA , inhibitor of tryptophan hydroxylase , P17752 ) . In addition , the effect of a low-tryptophan ( DB00150 ) diet was investigated . NSD 1015 or DB00150 depletion did not affect any model parameters , but PCPA reduced [(11)C]5-HTP uptake , and the k3 . This was unexpected as NSD 1015 directly inhibits the enzyme converting [(11)C]5-HTP to [(11)C]5-HT , suggesting that trapping of radioactivity does not distinguish between parent tracer and its metabolites . As different results have been acquired in monkeys and humans , [(11)C]5-HTP-PET may be suitable for measuring 5-HT synthesis in primates , but not in rodents . 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 . 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 . 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 . 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 . 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 . [ Granulopoietic inhibitor ] . The differentiation and proliferation of granulocytes are specifically supported by DB00099 . On the other hand , little is known about the negative modulation of granulopoiesis . We had found that the sera obtained from patients during the recovery phase of hematopoiesis after chemotherapy inhibited the formation of CFU-G , suggesting the modulation of granulopoiesis by negative regulator(s) . As the inhibitory effect was specific for CFU-G , the inhibitor in this inhibitory sera was suggested the soluble Q99062 . Several Q99062 fragments were detected in the concentrated fractionated-urine by the immunoblot analysis using the specific antibody against the cytokine receptor homologous ( P06850 ) domain of Q99062 . G- P04141 binding protein was successfully obtained from normal human plasma by G- P04141 affinity column . Major component was the 90Kd of protein which was thought to be the extracellular domain of Q99062 by the immunoblot analysis . About 90Kd- and 30Kd- bands of the binding-complex formation between G- P04141 monomer and the extracellular domain or the fragment of Q99062 were shown in the immunoblot analysis using anti-G- P04141 Ab . CFU-G formation was suppressed by the solution of soluble Q99062 fragments , suggesting the competition of G- P04141 which bind to granulocyte precursor cells . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . 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 . Identification and localization of DNA alteration in Chinese hamster ovary cell mutants ( Urd- ) defective in the first three enzymes of de novo pyrimidine synthesis . In animals , the first three enzymatic steps of de novo pyrimidine synthesis , carbamyl phosphate synthetase , aspartate transcarbamylase , and dihydroorotase , comprise the multifunctional protein known as the P27708 . Mutants of Chinese hamster ovary cells ( CHO- P04264 , pro- ) deficient in P27708 activities require uridine for growth and are designated Urd-A mutants . To examine further the nature of the genetic alterations in Urd-A mutants and revertants , we have performed a detailed Southern blot hybridization analysis of DNA from wild-type , Urd-A , and revertant cells using as hybridization probes cDNAs complementary to CAD mRNA isolated from Syrian hamster . This has allowed us to identify an apparent alteration in the CAD gene in DNA from Urd-A cells . This alteration is in a region of the gene which appears to correspond to the region of the protein which is hypersensitive to proteases and which seems to be altered in the mutants . Only one of the two CAD alleles present appears to be altered in this way . Study of certain revertants of Urd-A strongly suggests that in some cases reversion has occurred by amplification of the mutant CAD allele . Constitutive activity in gonadotropin receptors . Constitutively active mutants ( CAMs ) of gonadotropin receptors are , in general , rare conditions . DB00044 -choriogonadotropin receptor ( P22888 ) CAMs provoke the dramatic phenotype of familial gonadotropin-independent isosexual male-limited precocious puberty , whereas in females , there is not yet any identified phenotype . Only one isolated follicle-stimulating hormone receptor ( P23945 ) P62158 ( Asp567Gly ) has so far been detected in a single male patient , besides other P23945 weak CAMs linked to pregnancy-associated ovarian hyperstimulation syndrome or to impaired desensitization and internalization . Several animal models have been developed for studying enhanced gonadotropin action ; in addition to unraveling valuable new information about the possible phenotypes of isolated P23945 and P22888 CAMs in women , the information obtained from these mouse models has served multiple translational goals , including the development of new diagnostic and therapeutic targets as well as the prediction of phenotypes for mutations not yet identified in humans . Mutagenesis and computational studies have shed important information on the physiopathogenic mechanisms leading to constitutive activity of gonadotropin receptors ; a common feature in these receptor CAMs is the release of stabilizing interhelical interactions between transmembrane domains ( TMDs ) 3 and 6 leading to an increase , with respect to the wild-type receptor , in the solvent accessibility at the cytosolic extension of TMDs 3 , 5 , and 6 , which involves the highly conserved DB00142 / DB00128 - DB00125 - DB00135 / DB00150 sequence . In this chapter , we summarize the structural features , functional consequences , and mechanisms that lead to constitutive activation of gonadotropin receptor CAMs and provide information on pharmacological approaches that might potentially modulate gonadotropin receptor P62158 function . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . 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 .

MH_train_1528

MH_train_1528

MH_train_1528

interacts_with DB00623?

multiple_choice

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 . 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 . Cytokine release syndrome after blinatumomab treatment related to abnormal macrophage activation and ameliorated with cytokine-directed therapy . DB09052 is a P15391 /CD3-bispecific T-cell receptor-engaging ( BiTE ) antibody with efficacy in refractory B-precursor acute lymphoblastic leukemia . Some patients treated with blinatumomab and other T cell-activating therapies develop cytokine release syndrome ( CRS ) . We hypothesized that patients with more severe toxicity may experience abnormal macrophage activation triggered by the release of cytokines by T-cell receptor-activated cytotoxic T cells engaged by BiTE antibodies and leading to hemophagocytic lymphohistiocytosis ( HLH ) . We prospectively monitored a patient during blinatumomab treatment and observed that he developed HLH . He became ill 36 hours into the infusion with fever , respiratory failure , and circulatory collapse . He developed hyperferritinemia , cytopenias , hypofibrinogenemia , and a cytokine profile diagnostic for HLH . The HLH continued to progress after discontinuation of blinatumomab ; however , he had rapid improvement after P05231 receptor-directed therapy with tocilizumab . Patients treated with T cell-activating therapies , including blinatumomab , should be monitored for HLH , and cytokine-directed therapy may be considered in cases of life-threatening CRS . This trial was registered at www.clinicaltrials.gov as # NCT00103285 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . Exocytosis of insulin promotes insulin gene transcription via the insulin receptor/ P19957 kinase/ P08133 s6 kinase and P62158 kinase pathways . The control of glucose homeostasis by insulin requires , in addition to the glucose-induced insulin release , a highly dynamic control of insulin biosynthesis . Although elevated glucose concentrations have been shown to trigger insulin biosynthesis at the levels of transcription and translation , the molecular mechanisms underlying the immediate transcriptional control are poorly understood . By investigating signal transduction pathways involved in the " glucose-dependent " transcriptional control , thereby analyzing endogenous (prepro)insulin mRNA levels and monitoring on-line insulin promoter-driven GFP expression , we provide , for the first time , evidence that physiologically stimulated insulin secretion from the pancreatic beta cell promotes insulin biosynthesis by enhancing insulin gene transcription in an autocrine manner . We show that secreted insulin acts via beta-cell insulin receptors and up-regulates insulin gene transcription by signaling through the Q9Y4H2 / P19957 kinase/ P08133 s6k and P62158 kinase pathways . 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 . 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 . P01579 -induced P56817 expression is mediated by activation of O60674 and P27361 /2 signaling pathways and direct binding of P42224 to P56817 promoter in astrocytes . P56817 ( P56817 ) is an essential enzyme for the production of beta amyloid . Since we found that injection of interferon-gamma ( P01579 ) into young mouse brains increased P56817 expression in astrocytes , we investigated molecular mechanisms underlying this process by cloning a putative P56817 promoter . P56817 promoter activity was differentially regulated by P01579 in a region specific manner and down-regulated by an inhibitor of O60674 ( O60674 ) . A dominant negative mutant of signal transducer and activator of transcription 1 ( P42224 ) expression suppressed P56817 promoter activity , and this was rescued by transfecting wild type P42224 . Electrophoretic mobility shift assay and promoter activity assays indicated that P42224 binds directly to the putative P42224 binding sequence of P56817 promoter . Because P01579 treatment induced P42224 phosphorylation , we examined whether the expression of a suppressor of cytokine signaling ( Q9NSE2 ) , negative regulator of O60674 , suppresses P56817 promoter activity . The results show that O15524 or O14543 expression suppressed P56817 promoter by blocking phosphorylation of Tyr701 residue in P42224 . Also , because P01579 treatment specifically potentiated extracellular signal regulated Q96HU1 kinase ( P29323 ) 1/2 activation , pretreatment of mitogen-activated or extracellular signal-regulated protein kinase ( MEK ) inhibitor , PD98059 , significantly attenuated P01579 -induced P56817 promoter activity and protein expression through blocking phosphorylation of Ser727 residue in P42224 , suggesting that P27361 /2 is associated with P01579 -induced P42224 signaling cascade . Taken together , our results suggest that P01579 activates O60674 and P27361 /2 and then phosphorylated P42224 binds to the putative P42224 binding sequences in P56817 promoter region to modulate P56817 protein expression in astrocytes . DB01892 is a novel type of P09917 inhibitor with high efficacy in vivo . We previously showed that , in vitro , hyperforin from St . John 's wort ( Hypericum perforatum ) inhibits P09917 ( P09917 ) , the key enzyme in leukotriene biosynthesis . Here , we demonstrate that hyperforin possesses a novel and unique molecular pharmacological profile as a P09917 inhibitor with remarkable efficacy in vivo . DB01892 ( 4 mg/kg , i.p. ) significantly suppressed leukotriene B(4) formation in pleural exudates of carrageenan-treated rats associated with potent anti-inflammatory effectiveness . Inhibition of P09917 by hyperforin , but not by the iron-ligand type P09917 inhibitor BWA4C or the nonredox-type inhibitor ZM230487 , was abolished in the presence of phosphatidylcholine and strongly reduced by mutation ( W13A-W75A-W102A ) of the P09917 P06681 -like domain . Moreover , hyperforin impaired the interaction of P09917 with coactosin-like protein and abrogated P09917 nuclear membrane translocation in ionomycin-stimulated neutrophils , processes that are typically mediated via the regulatory P09917 P06681 -like domain . Together , hyperforin is a novel type of P09917 inhibitor apparently acting by interference with the P06681 -like domain , with high effectiveness in vivo . 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 . DB00360 supplementation reduces atherosclerosis and vascular inflammation in apolipoprotein E-knockout mice . BH4 ( tetrahydrobiopterin ) supplementation improves endothelial function in models of vascular disease by maintaining P29474 ( endothelial nitric oxide synthase ) coupling and NO ( nitric oxide ) bioavailability . However , the cellular mechanisms through which enhanced endothelial function leads to reduced atherosclerosis remain unclear . We have used a pharmaceutical BH4 formulation to investigate the effects of BH4 supplementation on atherosclerosis progression in ApoE-KO ( apolipoprotein E-knockout ) mice . Single oral dose pharmacokinetic studies revealed rapid BH4 uptake into plasma and organs . Plasma BH4 levels returned to baseline by 8 h after oral dosing , but remained markedly increased in aorta at 24 h . Daily oral BH4 supplementation in ApoE-KO mice from 8 weeks of age , for a period of 8 or 12 weeks , had no effect on plasma lipids or haemodynamic parameters , but significantly reduced aortic root atherosclerosis compared with placebo-treated animals . BH4 supplementation significantly reduced P19320 ( vascular cell adhesion molecule 1 ) mRNA levels in aortic endothelial cells , markedly reduced the infiltration of T-cells , macrophages and monocytes into plaques , and reduced T-cell infiltration in the adjacent adventitia , but importantly had no effect on circulating leucocytes . P30793 ( P30793 ) -transgenic mice , with a specific increase in endothelial BH4 levels , exhibited a similar reduction in vascular immune cell infiltration compared with BH4-deficient controls , suggesting that BH4 reduces vascular inflammation via endothelial cell signalling . In conclusion , BH4 supplementation reduces vascular immune cell infiltration in atherosclerosis and may therefore be a rational therapeutic approach to reduce the progression of atherosclerosis . 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 ) 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 .

MH_train_1529

MH_train_1529

MH_train_1529

interacts_with DB09053?

multiple_choice

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 . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . Direct intracerebral delivery of cintredekin besudotox ( P35225 -PE38QQR ) in recurrent malignant glioma : a report by the DB05305 Intraparenchymal Study Group . PURPOSE : Glioblastoma multiforme ( GBM ) is a devastating brain tumor with a median survival of 6 months after recurrence . Cintredekin besudotox ( CB ) is a recombinant protein consisting of interleukin-13 ( P35225 ) and a truncated form of Pseudomonas exotoxin ( PE38QQR ) . Convection-enhanced delivery ( DB01333 ) is a locoregional-administration method leading to high-tissue concentrations with large volume of distributions . We assessed the use of intracerebral DB01333 to deliver CB in patients with recurrent malignant glioma ( MG ) . PATIENTS AND METHODS : Three phase I clinical studies evaluated intracerebral DB01333 of CB along with tumor resection . The main objectives were to assess the tolerability of various concentrations and infusion durations ; tissue distribution ; and methods for optimizing delivery . All patients underwent tumor resection followed by a single intraparenchymal infusion ( in addition to the intraparenchymal one following resection ) , with a portion of patients who had a preresection intratumoral infusion . RESULTS : A total of 51 patients with MG were treated including 46 patients with GBM . The maximum tolerated intraparenchymal concentration was 0.5 microg/mL and tumor necrosis was observed at this concentration . Infusion durations of up to 6 days were well tolerated . Postoperative catheter placement appears to be important for optimal drug distribution . CB- and procedure-related adverse events were primarily limited to the CNS . Overall median survival for GBM patients is 42.7 weeks and 55.6 weeks for patients with optimally positioned catheters with patient follow-up extending beyond 5 years . CONCLUSION : CB appears to have a favorable risk-benefit profile . DB01333 is a complex delivery method requiring catheter placement via a second procedure to achieve accurate catheter positioning , better drug distribution , and better outcome . 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 . 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 . Inhibitors of P11274 signalling interrupt the survival signal mediated by the micro-environment in mantle cell lymphoma . Several studies provide evidences for mantle cell lymphoma ( Q8WXI8 ) cell survival relying on B-cell receptor ( P11274 ) -mediated signalling pathways , whereas the nature of this activation is unknown . Significant progress in Q8WXI8 treatment is achieved through therapies targeting P11274 -associated kinases , i.e. , DB09053 and Fostamatinib , inhibitors of Q06187 and P43405 , respectively . Our study addresses survival signals emanating from the P11274 or the tumour environment and how inhibiting P11274 signalling effectors might impact these survival signals . We found that Q06187 was constitutively activated and that P43405 phosphorylation was highly increased and sustained upon P11274 activation of primary Q8WXI8 cells . Moreover , Q8WXI8 cells from leukaemic patients secreted high amount of IL-1β , P05231 , P10145 and P13501 . Activation of the P11274 induced ( i ) cell survival , ( ii ) P40763 activation and ( iii ) increased autocrine secretion of IL-1β , P05231 , P10145 , P13501 , P22301 , TNFα and P15692 . Specific inhibition of Q06187 by DB09053 or P43405 by Fostamatinib ( R406 ) reversed these protective effects and decreased both basal and P11274 -induced autocrine cytokine secretions associated with P40763 phosphorylation . Interestingly , targeting Q06187 and P43405 prevented and inhibited P11274 -induced Q8WXI8 cell adhesion to human bone marrow stromal cells ( HMSCs ) in short- and long-term co-culture . We demonstrated that P11274 -induced survival relies on autocrine secretion of IL-1β , TNFα and P13501 that might facilitate adhesion of Q8WXI8 cells to HMSC . Treatment with DB09053 or Fostamatinib blocked the chemotactic signal thus increasing apoptosis . Xaliproden ( SR57746A ) induces P08908 receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT-1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . Evaluation of degarelix in the management of prostate cancer . Medical castration using gonadotropin-releasing hormone ( DB00644 ) receptor agonists currently provides the mainstay of androgen deprivation therapy for prostate cancer . Although effective , these agents only reduce testosterone levels after a delay of 14 to 21 days ; they also cause an initial surge in testosterone that can stimulate the cancer and lead to exacerbation of symptoms ( " clinical flare " ) in patients with advanced disease . Phase III trial data for the recently approved P30968 blocker , degarelix , demonstrated that it is as effective and well tolerated as DB00644 agonists . However , it has a pharmacological profile more closely matching orchiectomy , with an immediate onset of action and faster testosterone and PSA suppression , without a testosterone surge or microsurges following repeated injections . As a consequence , with this DB00644 blocker , there is no risk of clinical flare and no need for concomitant antiandrogen flare protection . DB06699 therefore provides a useful addition to the hormonal armamentarium for prostate cancer and offers a valuable new treatment option for patients with hormone-sensitive advanced disease . Here , we review key preclinical and clinical data for degarelix , and look at patient-focused perspectives in the management of prostate cancer . DB09079 , a triple angiokinase inhibitor , enhances cytotoxic therapy response in pancreatic cancer . Angiogenesis remains a sensible target for pancreatic ductal adenocarcinoma ( PDAC ) therapy . P15692 , PDGF , FGF and their receptors are expressed at high levels and correlate with poor prognosis in human PDAC . DB09079 is a triple angiokinase inhibitor that targets P17948 /2/3 , P11362 /2/3 and PDGFRα/β signaling . We investigated the antitumor activity of nintedanib alone or in combination with the cytotoxic agent gemcitabine in experimental PDAC . DB09079 inhibited proliferation of cells from multiple lineages found in PDAC , with gemcitabine enhancing inhibitory effects . DB09079 blocked PI3K/MAPK activity and induced apoptosis in vitro and in vivo . In a heterotopic model , net local tumor growth compared to controls ( 100 % ) was 60.8 ± 10.5 % in the gemcitabine group , -2.1 ± 9.9 % after nintedanib therapy and -12.4 ± 16 % after gemcitabine plus nintedanib therapy . Effects of therapy on intratumoral proliferation , microvessel density and apoptosis corresponded with tumor growth inhibition data . In a PDAC survival model , median animal survival after gemcitabine , nintedanib and gemcitabine plus nintedanib was 25 , 31 and 38 days , respectively , compared to 16 days in controls . The strong antitumor activity of nintedanib in experimental PDAC supports the potential of nintedanib-controlled mechanisms as targets for improved clinical PDAC therapy . DB06016 , an orally active D2/D3 receptor antagonist , for the potential treatment of schizophrenia , bipolar mania and depression . DB06016 ( RGH-188 ) , which is being codeveloped by Gedeon Richter Ltd , Forest Laboratories Inc and Mitsubishi Tanabe Pharma Corp , is a novel putative antipsychotic drug that exerts partial agonism at dopamine D2/D3 receptors , with preferential binding to D3 receptors , and partial agonism at serotonin P08908 receptors . Its activity at D2/D3 receptors may be lower than that of the prototype partial agonist aripiprazole . The antipsychotic activity of cariprazine was demonstrated in animal models , and data also suggest that the propensity for extrapyramidal side effects is low and that the drug may have procognitive properties . DB06016 is rapidly absorbed , with high oral bioavailability and a long plasma elimination t1/2 . DB06016 is in phase III clinical trials in patients with schizophrenia and in patients with bipolar disorder . Data from phase II trials in patients with schizophrenia and bipolar mania indicate that the drug has antipsychotic and antimanic properties that are superior to placebo . With its unique receptor affinity profile , cariprazine may represent a potential enrichment of the therapeutic armamentarium for schizophrenia and affective disorders . Its activity against the cognitive deficits associated with schizophrenia has to be carefully investigated . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . Clonal amniotic fluid-derived stem cells express characteristics of both mesenchymal and neural stem cells . Recent evidence has shown that amniotic fluid may be a novel source of fetal stem cells for therapeutic transplantation . We previously developed a two-stage culture protocol to isolate a population of amniotic fluid-derived DB05914 ( AFMSCs ) from second-trimester amniocentesis . AFMSCs maintain the capacity to differentiate into multiple mesenchymal lineages and neuron-like cells . It is unclear whether amniotic fluid contains heterogeneous populations of stem cells or a subpopulation of primitive stem cells that are similar to marrow stromal cells showing the behavior of neural progenitors . In this study , we showed a subpopulation of amniotic fluid-derived stem cells ( AF-SCs ) at the single-cell level by limiting dilution . We found that Q9H9S0 - and Q01860 ( also known as Q01860 ) -expressing cells still existed in the expanded single cell-derived AF-SCs . Aside from the common mesenchymal characteristics , these clonal AF-SCs also exhibit multiple phenotypes of neural-derived cells such as P48681 , Q13509 , P12036 , NEUNA60 , P54803 , and P14136 expressions both before and after neural induction . Most importantly , HPLC analysis showed the evidence of dopamine release in the extract of dopaminergic-induced clonal AF-SCs . The results of this study suggest that besides being an easily accessible and expandable source of fetal stem cells , amniotic fluid will provide a promising source of neural progenitor cells that may be used in future cellular therapies for neurodegenerative diseases and nervous system injuries . 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 . [ Gene expression profiling using oligonucleotide microarray in atrophic gastritis and intestinal metaplasia ] . BACKGROUND/AIMS : The atrophic gastritis with intestinal metaplasia of gastric mucosa has been considered to be the major factor of carcinogenesis in the stomach . However , the key molecules are still poorly understood . To elucidate the molecular genetic basis , we report the results of our initial microarray data to analyze the genome pattern in patients with atrophic gastritis and intestinal metaplasia of the stomach . METHODS : We used oligonucleotide microarray technique to evaluate the gene expression profiles in atrophic gastritis with intestinal metaplasia , in comparison with those of normal mucosa . For the identification of differentially expressed genes , Significance Analysis of Microarrays ( DB00118 ) package method was used . The results were analyzed using global normalization , intensity dependent normalization , and box plot normalization . RESULTS : Eight genes including FABP , P05451 , Q96RD1 , MEP1 , P30531 , SI , Mucin 1 , and Q9ULC3 in mucosa of atrophic gastritis and intestinal metaplasia were up-regulated by more than 10 times as compared with normal gastric mucosa . Only one gene , LOC44119 was down-regulated by more than 10 times of the expression as compared with normal gastric mucosa . In respect to the expression of known genes related to gastric carcinogenesis , 8 genes including P02751 , Q9H3Y6 , P04637 , TP53IMP2 , Q53FA7 , P22455 , P01137 , and P01135 showed up- and down-regulations more than 2 folds in expression pattern . CONCLUSIONS : We could identify a total genome pattern in patient with atrophic gastritis and intestinal metaplasia using oligonucleotide microarray . We believe that the current results will serve as a fundamental bioinformative basis for clinical applications in diagnosis and treatment of gastric cancer and precancerous lesion in the future . Gα(olf) mutation allows parsing the role of DB02527 -dependent and extracellular signal-regulated kinase-dependent signaling in DB01235 -induced dyskinesia . Although DB01235 ( DB01235 ) remains the reference treatment of Parkinson 's disease , its long-term beneficial effects are hindered by DB01235 -induced dyskinesia ( LID ) . In the dopamine ( DA ) -denervated striatum , DB01235 activates DA D₁ receptor(D₁R) signaling , including DB02527 -dependent protein kinase A ( PKA ) and extracellular signal-regulated kinase ( P29323 ) , two responses associated with LID . However , the cause of PKA and P29323 activation , their respective contribution to LID , and their relationship are not known . In striatal neurons , D₁R activates adenylyl-cyclase through Gα(olf) , a protein upregulated after lesion of DA neurons in rats and inpatients . We report here that increased Gα(olf) levels in hemiparkinsonian mice are correlated with LID after chronic DB01235 treatment . To determine the role of this upregulation , we performed unilateral lesion in mice lacking one allele of the Gnal gene coding for Gα(olf) ( Gnal⁺/⁻ ) . Despite an increase in the lesioned striatum,Gα(olf) levels remained below those of unlesioned wild-type mice . In Gnal⁺/⁻ mice , the lesion-induced DB01235 stimulation of DB02527 /PKA-mediated phosphorylation of P42261 Ser845 and Q9UD71 ( 32 kDa DA- and DB02527 -regulated phosphoprotein ) Thr34 was dramatically reduced , whereas P29323 activation was preserved . LID occurrence was similar in Gnal⁺/⁺ and Gnal⁺/⁻ mice after a 10-d DB01235 ( 20 mg/kg ) treatment . Thus , in lesioned animals , Gα(olf) upregulation is critical for the activation by DB01235 of D₁R-stimulated DB02527 /PKA but not P29323 signaling . Although the DB02527 /PKA pathway appears to be required for LID development , our results indicate that its activation is unlikely to be the main source of LID . In contrast , the persistence of DB01235 -induced P29323 activation in Gnal⁺/⁻ mice supports its causal role in LID development . Q06187 is a therapeutic target in stem-like cells from multiple myeloma . DB09053 ( Imbruvica ) , a small-drug inhibitor of Q06187 ( Q06187 ) , is currently undergoing clinical testing in patients with multiple myeloma , yet important questions on the role of Q06187 in myeloma biology and treatment are outstanding . Using flow-sorted side population cells from human myeloma cell lines and multiple myeloma primary samples as surrogate for the elusive multiple myeloma stem cell , we found that elevated expression of Q06187 in myeloma cells leads to AKT/WNT/β-catenin-dependent upregulation of key stemness genes ( Q01860 , P48431 , Q9H9S0 , and MYC ) and enhanced self-renewal . Enforced transgenic expression of Q06187 in myeloma cells increased features of cancer stemness , including clonogenicity and resistance to widely used myeloma drugs , whereas inducible knockdown of Q06187 abolished them . Furthermore , overexpression of Q06187 in myeloma cells promoted tumor growth in laboratory mice and rendered side population-derived tumors that contained high levels of Q06187 more sensitive to the selective , second-generation Q06187 inhibitor , CGI1746 , than side population-derived tumors that harbored low levels of Q06187 . Taken together , these findings implicate Q06187 as a positive regulator of myeloma stemness and provide additional support for the clinical testing of Q06187 -targeted therapies in patients with myeloma . DB00644 induction of extracellular-signal regulated kinase is blocked by inhibition of calmodulin . Our previous studies demonstrate that DB00644 -induced P29323 activation required influx of extracellular Ca2+ in alphaT3-1 and rat pituitary cells . In the present studies , we examined the hypothesis that calmodulin ( Cam ) plays a fundamental role in mediating the effects of Ca2+ on P29323 activation . Cam inhibition using W7 was sufficient to block DB00644 -induced reporter gene activity for the c-Fos , murine glycoprotein hormone alpha-subunit , and MAPK phosphatase ( MKP ) -2 promoters , all shown to require P29323 activation . Inhibition of Cam ( using a dominant negative ) was sufficient to block DB00644 -induced P29323 but not c-Jun N-terminal kinase activity activation . The Cam-dependent protein kinase ( CamK ) II inhibitor KN62 did not recapitulate these findings . DB00644 -induced phosphorylation of Q02750 and c-Raf kinase was blocked by Cam inhibition , whereas activity of phospholipase C was unaffected , suggesting that Ca2+/Cam modulation of the P29323 cascade potentially at the level of c-Raf kinase . Enrichment of Cam-interacting proteins using a Cam agarose column revealed that c-Raf kinase forms a complex with Cam . Reconstitution studies reveal that recombinant c-Raf kinase can associate directly with Cam in a Ca2+-dependent manner and this interaction is reduced in vitro by addition of W7 . Cam was localized in lipid rafts consistent with the formation of a Ca2+-sensitive signaling platform including the P30968 and c-Raf kinase . These data support the conclusion that Cam may have a critical role as a Ca2+ sensor in specifically linking Ca2+ flux with P29323 activation within the DB00644 signaling pathway . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . Q08881 inhibitors in inflammation and immune-mediated disorders . P60568 -inducible T cell kinase ( Q08881 ) is a non-receptor tyrosine kinase expressed in T cells , NKT cells and mast cells which plays a crucial role in regulating the T cell receptor ( TCR ) , P10747 , P06729 , chemokine receptor P61073 , and FcepsilonR-mediated signaling pathways . In T cells , Q08881 is an important mediator for actin reorganization , activation of PLCgamma , mobilization of calcium , and activation of the NFAT transcription factor . Q08881 plays an important role in the secretion of P60568 , but more critically , also has a pivotal role in the secretion of Th2 cytokines , P05112 , P05113 and P35225 . As such , Q08881 has been shown to regulate the development of effective Th2 response during allergic asthma as well as infections of parasitic worms . This ability of Q08881 to regulate Th2 responses , along with its pattern of expression , has led to the proposal that it would represent an excellent target for Th2-mediated inflammation . We discuss here the possibilities and pitfalls of targeting Q08881 for inflammatory disorders . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . 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 .

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interacts_with DB00864?

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Hyponatremia associated with DB06186 -induced hypophysitis . A 75-year-old woman with a history of stage IV metastatic melanoma underwent treatment with the P16410 blocking agent DB06186 . She presented 2 months after initiating treatment with a severe headache . Laboratories were consistent with severe hyponatremia . Q9BWK5 of the brain revealed enlargement of the pituitary gland , enhancement of the infundibulum , and an enhancing , centrally necrotic foci in the anterior pituitary . Based on the clinical and radiographic findings , she was diagnosed with treatment-related syndrome of inappropriate antidiuretic hormone secretion ( SIADH ) . Effective treatment consisted of fluid restriction , hyperosmolar therapy , and steroids . 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 . 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 . Anti-inflammatory effect of parenteral fish oil lipid emulsion on human activated mononuclear leukocytes . BACKGROUND & AIM : To compare the effect of fish oil-based ( FO ) lipid emulsions ( LE ) for parenteral administration with standard LE and a new FO containing LE composed of four different oils on the antigen presentation and inflammatory variables . METHODS : Phytohemagglutinin ( PHA ) activated human mononuclear leukocytes were cultured with different LE - Control : without LE ; SO : soybean oil ; SO/FO : soybean and FO ( 4:1 ) ; Q8IVS2 /SO : medium chain triglycerides and SO ( 1:1 ) ; Q8IVS2 /SO/FO : Q8IVS2 /SO and FO ( 4:1 ) and SMOF : a new LE containing FO . Cytokine production was evaluated by ELISA , the expression of antigen-presenting and co-stimulatory surface molecules were analyzed by flow cytometry and lymphocyte proliferation was assessed by H(3)- DB04485 incorporation , after tetanus toxoid-induced activation . RESULTS : All LE decreased the HLA-DR and increased P10747 and P16410 expression on monocytes/macrophages and lymphocytes surface ( p < 0.05 ) . SO/FO and Q8IVS2 /SO/FO decreased lymphocyte proliferation ( p < 0.05 ) . All LE decreased P60568 production , but this effect was enhanced with Q8IVS2 /SO/FO and SMOF ( p < 0.05 ) . Q8IVS2 /SO/FO decreased P05231 and increased P22301 , whereas SO had the opposite effect ( p < 0.05 ) . CONCLUSION : FO LE inhibited lymphocyte proliferation and had an anti-inflammatory effect . These effects seem to be enhanced when FO is mixed with Q8IVS2 /SO . SMOF had a neutral impact on lymphocyte proliferation and P05231 and P22301 production . T cells stimulated by P29965 positive leukemic blasts-pulsed dendritic cells meet optimal functional requirements for adoptive T-cell therapy . Adoptive T-cell immunotherapy may provide complementary therapy for childhood B-cell precursor acute lymphoblastic leukemia ( P03999 -ALL ) . In this study , we have analyzed the functional characteristics of anti- P03999 -ALL effector T cells generated by co-culturing T lymphocytes and dendritic cells ( DC ) from allogeneic human stem cell transplantation ( HSCT ) donors . After 21-day co-culture with DC pulsed with P29965 + apoptotic P03999 -ALL blasts , T cells presented with both effector and central memory phenotype , and showed high and specific cytotoxic activity against leukemic cells ( average lysis = 77 % ) , mostly mediated by CD8+ T cells . Noticeably , growth of P01730 T cells was maintained ( 45 % of total cells ) , which actively produced Th1 cytokines ( P01579 , P01375 , P60568 ) , but not P05112 , P05113 and P22301 . Anti- P03999 -ALL T cells expressed CD49d and P61073 ( implicated in the recruitment to bone marrow ) , and CD62L and P32248 ( involved in the migration to lymphoid organs ) . In accordance with this profile , T cells significantly migrated in response to the chemokines P48061 and Q99731 . In conclusion , stimulation of T cells with P29965 + P03999 -ALL cells-loaded DC not only elicited the generation of potent and specific anti-leukemic cytotoxic effectors , but also the differentiation of specific and functional Th-1 P01730 lymphocytes . These effectors are fully equipped to reach leukemia-infiltrated tissues and have characteristics to support and orchestrate the anti-tumor immune-response . DB00864 ( FK506 ) increases neuronal expression of P20936 -43 and improves functional recovery after spinal cord injury in rats . DB00864 ( FK506 ) , a widely used immunosuppressant drug , has neurite-promoting activity in cultured PC12 cells and peripheral neurons . The present study investigated whether tacrolimus affects the expression of the neuronal growth-associated protein , P20936 -43 , as well as functional recovery after photothrombotic spinal cord injury in the rat . In injured animals receiving tacrolimus , the number of neurons expressing P20936 -43 mRNA and protein approximately doubled compared to that in injured animals receiving vehicle alone . This increase in P20936 -43-positive cells was paralleled by a significant improvement in neurological function evaluated by open-field and inclined plane tests . Another P62942 ligand ( V-10,367 ) had similar effects on P20936 -43 expression and functional outcome , indicating that the observed effects of tacrolimus do not involve inhibition of the phosphatase calcineurin . Thus , tacrolimus , a drug which is already approved for use in humans , as well as other P62942 ligands which do not inhibit calcineurin , could potentially enhance functional outcome after CNS injury in humans . Forced dimerization increases the activity of Δ P00533 /EGFRvIII and enhances its oncogenicity . Delta epidermal growth factor receptor ( Δ P00533 ) , an in-frame deletion mutant of the extracellular ligand-binding domain , which occurs in about 30 % of glioblastoma , is a potent oncogene that promotes tumor growth and progression . The signaling of Δ P00533 is ligand-independent and low intensity , allowing it to evade the normal mechanisms of internalization and degradation by the endocytic machinery and hence is persistent . The basis of the oncogenic potential of Δ P00533 remains incompletely understood , including whether dimerization plays an important role in its signal and whether its oncogenic potential is dependent on its relatively low intensity , when compared with the acutely activated wild-type receptor . To examine these two important questions , we have generated a chimeric Δ P00533 that allows forced dimerization via domains derived from variants of the P62942 protein that are brought together by FK506 derivatives . Forced dimerization of chimeric Δ P00533 significantly increased the intensity of its signal , as measured by receptor phosphorylation levels , suggesting that the naturally occurring Δ P00533 does not form strong or stable dimers as part of its low level signal . Interestingly , the increased activity of dimerized , chimeric Δ P00533 did not promote receptor internalization , implying that reduced rate of endocytic downregulation of Δ P00533 is an inherent characteristic . Significantly , forced dimerization enhanced the oncogenic signal of the receptor , implying that the Δ P00533 is a potent oncogene despite , not because of its low intensity . Distribution of immunophilin P62942 protein and mRNA within the mammalian cochlea and cochlear nucleus . Immunophilin FK binding protein-12 ( P62942 ) , the soluble receptor for the immunosuppressant drug FK506 , is involved in a number of neuronal activities including increased nerve regeneration in the peripheral nervous system and enhanced recovery in animal models of neurodegenerative diseases . In addition , P62942 is tightly bound to the calcium release channel ryanodine receptor and physiologically interacts with the inositol 1,4,5-trisphosphate receptor . In nearly all cell types , release of intracellular Ca(2+) and subsequent second messenger signaling involves activation of these ion channels . We determined the distribution of P62942 within the mammalian cochlea and dorsal cochlear nucleus ( P07585 ) in order to gain insight into Ca(2+) regulation within the cochlea and to possibly identify potential cellular targets for neuroimmunophilin ligands that may prove useful in protection and recovery following ototoxic insult . P62942 protein and mRNA were found to be abundant throughout rat and guinea pig cochlea and P07585 . 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 . DB01169 inhibits osteosarcoma cell invasiveness via MAPK signaling pathway . DB01169 ( As(2)O(3) ) is an active ingredient in traditional Chinese medicine . Recent studies showed that it causes apoptosis in several cancer cells . However , research of As(2)O(3) in osteosarcoma is sparse . In our present study , an inhibitory effect of As(2)O(3) on osteosarcoma cell adhesion and metastasis was observed with a cell adhesion , migration and invasion test . The impact of As(2)O(3) on the activities of P14780 and MAPK pathway-related downstream factors was analyzed by western blotting . Our results showed that As(2)O(3) significantly inhibited motility , migration and invasion in Q9UKB1 and MNNG cells in a concentration-dependent manner at concentrations ranging from 0.5-2 μM , and led to cytoskeletal rearrangements . As(2)O(3) exerted an inhibitory effect on the phosphorylation of P27361 /2 and MEK , which are the members of the MAPK family . Additionally , treatment with As(2)O(3) in combination with inhibitors specific for MEK ( U0126 ) in Q9UKB1 and MNNG cells resulted in a marked inhibition of cell invasion and As(2)O(3) could significantly reduce PMA-induced invasion . In conclusion , we demonstrate the inhibitory effects of As(2)O(3) on the invasiveness of Q9UKB1 and MNNG cells , which may be due at least partly to inactivation of the MAPK signaling pathway . Association of DB00135 phosphorylation of P20936 with mitogenic action of serotonin . We have previously shown that serotonin ( 5-HT ) induces both hyperplasia and hypertrophy of pulmonary artery smooth muscle cells ( SMC ) but not of endothelial cells ( EC ) through its high-affinity uptake . The present studies demonstrate rapid enhancement by 5-HT of DB00135 phosphorylation of proteins , including p120 , which also occurs in SMC but not in EC . The p120 protein was identified as P20936 ( P20936 ) by immunoprecipitation . Its phosphorylation occurred within minutes and preceded other events associated with 5-HT-induced mitogenesis . DB00135 kinase ( TK ) and 5-HT uptake inhibitors and 8-bromoadenosine 3',5'-cyclic monophosphate blocked both the 5-HT-induced DNA synthesis and DB00135 phosphorylation of P20936 . Vanadate elevated DNA synthesis and DB00135 phosphorylation of P20936 of both control and 5-HT-treated cells . 5-HT failed to alter DB00135 phosphorylation of P20936 in cellular homogenates , as opposed to intact cells . In the presence of DB07954 , 5-HT inhibited cellular growth , presumably through its action on P08908 or Q13639 receptors and elevation of adenosine 3',5'-cyclic monophosphate , but this was not associated with an alteration of DB00135 phosphorylation of P20936 . Similarly , a 5-HT1 or 5-HT2 receptor agonist failed to stimulate DB00135 phosphorylation or DNA synthesis of SMC . Stimulation of cellular proliferation and enlargement produced by 1 microM 5-HT were totally abolished by TK inhibitors that did not affect 5-HT uptake . These data indicate that DB00135 phosphorylation of P20936 may act as an intermediate signal in 5-HT-induced mitogenesis of SMC which requires cellular internalization of 5-HT rather than its action on a membrane receptor . Significant reduction of granulomas in Nrf2-deficient mice infected with Mycobacterium tuberculosis . OBJECTIVE : We have reported previously that mice deficient in nuclear erythroid 2 P29466 -related factor 2 ( Nrf2 ) , which regulates the expression of antioxidant and detoxification genes , showed significant susceptibility to airway inflammatory responses when exposed to diesel exhaust particles for eight weeks . As disruption of Nrf2 promotes immune cells that stimulate Th2-like immunoresponsiveness , Nrf2-deficient mice may be resistant to M. tuberculosis infection . SETTING : Nrf2-deficient mice were infected with M. tuberculosis aerially , and the size of their granulomas and cytokine mRNA expression were compared with those of wild-type mice . RESULTS : Significant reduction of granuloma formation and tubercle bacilli in granulomas was noted in the deficient mice 27 weeks after infection , concurrently with higher expression of P60568 and P35225 mRNA . CONCLUSION : It is concluded that Nrf2 inversely regulates M. tuberculosis-induced granuloma development at the late stage . Cross talk between Ca2+ and redox signalling cascades in muscle and neurons through the combined activation of ryanodine receptors/Ca2+ release channels . DB01373 release mediated by the ryanodine receptors ( RyR ) Ca2+ release channels is required for muscle contraction and contributes to neuronal plasticity . In particular , Ca2+ activation of RyR-mediated Ca2+ release can amplify and propagate Ca2+ signals initially generated by Ca2+ entry into cells . Redox modulation of RyR function by a variety of non-physiological or endogenous redox molecules has been reported . The effects of RyR redox modification on Ca2+ release in skeletal muscle as well as the activation of signalling cascades and transcription factors in neurons will be reviewed here . Specifically , the different effects of S-nitrosylation or S-glutathionylation of RyR cysteines by endogenous redox-active agents on the properties of skeletal muscle RyRs will be discussed . Results will be presented indicating that these cysteine modifications change the activity of skeletal muscle RyRs , modify their behaviour towards both activators and inhibitors and affect their interactions with P62942 and calmodulin . In the hippocampus , sequential activation of P27361 /2 and CREB is a requisite for Ca2+-dependent gene expression associated with long-lasting synaptic plasticity . The effects of reactive oxygen/nitrogen species on RyR channels from neurons and RyR-mediated sequential activation of neuronal P27361 /2 and CREB produced by hydrogen peroxide and other stimuli will be discussed as well . 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 . Light and X-ray scattering show decorin to be a dimer in solution . P07585 is a widely distributed member of the extracellular matrix small leucine-rich repeat glycoprotein/proteoglycan family . For investigation of its physical properties , decorin from two sources ( young steer skin and a recombinant adenovirus ) was used . The first sample was extracted into 7 m urea and purified , while the second was isolated from medium conditioned by 293A cells infected with adenovirus and purified without chaotropes . The only chemical differences detected between these materials were a slightly shorter glycosaminoglycan chain and the retention of the propeptide on the latter . Circular dichroism spectra of the two samples were virtually identical , showing a high proportion of beta-sheet and beta-turn and little alpha-helix . The protein cores were completely denatured in 2.25 m guanidine HCl ( GdnHCl ) but recovered their secondary structure on removal of chaotrope . Light scattering of material eluted from gel-filtration columns in DB03754 -buffered saline , pH 7.0 , gave molecular mass values of 165 +/- 1 kDa and 84.6 +/- 4 kDa for intact decorin and the glycoprotein core produced by digestion with chondroitin ABC lyase , respectively . Intact recombinant prodecorin had a mass of 148 +/- 18 kDa . These values , which are double those estimated from SDS gel electrophoresis or from the known sequences and compositions , were halved in 2.5 m GdnHCl . Data from solution x-ray scattering of intact decorin and its core in DB03754 -buffered saline are consistent with a dimeric particle whose protein component has a radius of gyration of 31.6 +/- 0.4 A , a maximum diameter of 98 +/- 5 A , and approximates two intertwined C shapes . The complex of FK506-binding protein 12 and FK506 inhibits calcineurin phosphatase activity and IgE activation-induced cytokine transcripts , but not exocytosis , in mouse mast cells . FK506 and cyclosporin A ( DB00091 ) are immunosuppressive agents that inhibit P60568 production by activated T cells , but only DB00091 inhibits IgE activation-induced cytokine transcripts in mouse P08700 -dependent , bone marrow-derived mast cells ( BMMC ) . We previously associated the resistance of BMMC to FK506 with a deficiency in the expression of FK506 binding protein ( FKBP ) 12 , a molecule that forms a complex with FK506 capable of inhibiting calcineurin phosphatase activity in vitro . In this report , we establish that P62942 mediates FK506 inhibition of both calcineurin phosphatase activity and IgE activation-induced cytokine transcripts in a Kirsten murine sarcoma virus-immortalized mast cell line that is P62942 deficient . Overexpression of P62942 by transfection enhanced the ability of FK506 to inhibit calcineurin phosphatase activity ( IC50 = 2 nM ) , compared with cells transfected with the expression vector alone ( IC50 > 30 nM ) . The IC50 value for FK506 inhibition of IgE activation-induced transcripts for P01375 decreased from 40 nM in vector control cells to 10 nM in P62942 transfectants . Similarly , the IC50 value for inhibition of P05231 transcripts decreased from > 1000 nM in vector control cells to 35 nM in P62942 transfectants . In contrast , activation-elicited release of the secretory granule mediator beta-hexosaminidase was only partially inhibited by FK506 at 1000 nM , regardless of the levels of P62942 expressed by the cells . Thus , P62942 is the dominant cytosolic protein that mediates FK506 inhibition of P01375 and P05231 transcripts . Rindopepimut , a 14-mer injectable peptide vaccine against EGFRvIII for the potential treatment of glioblastoma multiforme . Celldex Therapeutics is developing rindopepimut ( DB05374 ) , a 14-mer injectable peptide vaccine for the potential treatment of glioblastoma multiforme ( GBM ) . Rindopepimut specifically targets a novel junctional epitope of the P00533 deletion mutant EGFRvIII , which is a constitutively active receptor that is expressed in approximately 60 to 70 % of patients with GBM . EGFRvIII expression is correlated with worse prognosis and reduced overall survival . Importantly , EGFRvIII is not expressed in normal brain tissue , making it an excellent therapeutic target . Preclinical studies demonstrated lasting tumor regression and increased survival times , as well as efficient generation of EGFRvIII-specific humoral and cellular immune responses , in animals expressing EGFRvIII and vaccinated with rindopepimut . Phase I and II clinical trials in patients with GBM demonstrated significantly increased median time to progression and overall survival time in those vaccinated with rindopepimut compared with matched historical controls . Only limited side effects have been observed in patients . Given these results , rindopepimut is an extremely promising therapy for patients with GBM . Phase I and II clinical trials in patients with GBM were ongoing at the time of publication . In the future , it may be beneficial to combine rindopepimut with other treatment modalities to further prolong survival . Cell and molecular determinants of in vivo efficacy of the BH3 mimetic DB05764 against pediatric acute lymphoblastic leukemia xenografts . PURPOSE : Predictive biomarkers are required to identify patients who may benefit from the use of BH3 mimetics such as DB05764 . This study investigated the efficacy of DB05764 against a panel of patient-derived pediatric acute lymphoblastic leukemia ( ALL ) xenografts and utilized cell and molecular approaches to identify biomarkers that predict in vivo DB05764 sensitivity . EXPERIMENTAL DESIGN : The in vivo efficacy of DB05764 was tested against a panel of 31 patient-derived ALL xenografts composed of Q03164 - , P03999 - , and T-ALL subtypes . Basal gene expression profiles of ALL xenografts were analyzed and confirmed by quantitative RT-PCR , protein expression and BH3 profiling . An in vitro coculture assay with immortalized human mesenchymal cells was utilized to build a predictive model of in vivo DB05764 sensitivity . RESULTS : DB05764 demonstrated impressive activity against pediatric ALL xenografts , with 19 of 31 achieving objective responses . Among P10415 family members , in vivo DB05764 sensitivity correlated best with low Q07820 mRNA expression levels . BH3 profiling revealed that resistance to DB05764 correlated with mitochondrial priming by Q13794 peptide , suggesting a functional role for Q07820 protein . Using an in vitro coculture assay , a predictive model of in vivo DB05764 sensitivity was built . Testing this model against 11 xenografts predicted in vivo DB05764 responses with high sensitivity ( 50 % ) and specificity ( 100 % ) . CONCLUSION : These results highlight the in vivo efficacy of DB05764 against a broad range of pediatric ALL subtypes and shows that a combination of in vitro functional assays can be used to predict its in vivo efficacy . 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 . DB01411 inhibits renal epithelial cyst progression via activation of AMP-activated protein kinase . Q9Y271 ( CysLT1 receptor ) antagonists were found to inhibit chloride secretion in human airway epithelial cells . Since chloride secretion in renal epithelial cells , which shares common mechanisms with airway epithelial cells , plays important roles in renal cyst progression in polycystic kidney disease ( Q15139 ) , this study was aimed to investigate effects of drugs acting as CysLT1 receptor antagonists on renal cyst progression and its underlying mechanisms . Effects of CysLT1 receptor antagonists on renal cyst growth and formation were determined using Madine Darby canine kidney ( MDCK ) cyst models . Mechanisms of actions of CysLT1 receptor antagonists were determined using short-circuit current measurement , assays of cell viability and cell proliferation , and immunoblot analysis of signaling proteins . Of the three drugs acting as CysLT1 receptor antagonists ( montelukast , pranlukast and zafirlukast ) tested , pranlukast was the most promising drug that inhibited MDCK cyst growth and formation without affecting cell viability . Its effect was independent of the inhibition of CysLT1 receptors . Instead , it reduced DB02527 -activated chloride secretion and proliferation of MDCK cells in an AMP-activated protein kinase ( AMPK ) -dependent manner and had no effect on P13569 protein expression . Interestingly , pranlukast enhanced AMPK activation via calcium/calmodulin-dependent protein kinase kinase beta ( CaMKKβ ) with consequent activation of acetyl- DB01992 carboxylase ( ACC ) and suppression of mammalian target of rapamycin ( P42345 ) pathway . These results indicate that pranlukast retards renal epithelial cyst progression by inhibiting DB02527 -activated chloride secretion and cell proliferation via CaMKKβ-AMPK- P42345 pathway . Therefore , pranlukast represents a class of known drugs that may have potential utility in Q15139 treatment . Inactivation of caspase-1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin-1 ( IL-1 ) -beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL-1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL-1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE/caspase-1 ) or through caspase-1 gene deletion . METHODS : P29466 was selectively blocked by using pralnacasan or DB05507 . IL-1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL-1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 inhibition reduced the release of IL-1beta in organotypic slices exposed to LPS+ DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 ( intraperitoneal , 25-200 mg/kg ) to rats blocked seizure-induced production of IL-1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase-1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase-1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL-1beta . Pharmacological investigation of the role of leukotrienes in the pathogenesis of experimental NSAID gastropathy . The role of leukotrienes in the pathogenesis of acute gastric ulceration induced by nonsteroidal antiinflammatory drugs was investigated using a rat model . One part of the study involved oral pretreatment with a leukotriene synthesis inhibitor 1 h prior to administration of indomethacin ( 20 mg/kg per os ) . Three hours after indomethacin , the extent of macroscopically visible gastric damage was determined , and gastric LTB4 synthesis was determined . The compounds tested were PF-5901 , A-64077 , nordihydroguaiaretic acid , and L-698,037 . Each compound produced dose-related inhibition of gastric LTB4 synthesis and a parallel reduction in the severity of indomethacin-induced damage . The antioxidant properties of these compounds was assessed using an in vitro assay . There was no correlation between the antioxidant properties of the compounds and their ability to reduce the severity of indomethacin-induced gastric damage . In the second part of the study , the effects of intravenous , administration of LTD4 and LTB4 receptor antagonists on indomethacin-induced gastric epithelial damage ( measured by permeability to [51Cr] DB00974 ) were assessed . The two Q9Y271 antagonists ( MK-571 and DB00549 ) significantly reduced the permeability changes induced by indomethacin , while the two LTB4 antagonists ( SC-41930 and LY-255,283 ) were without significant effect . Despite the reduction of gastric epithelial injury , blockade of LTD4 receptors did not markedly affect the extent of macroscopically visible injury . These data are consistent with the hypothesis that leukotrienes contribute to the epithelial injury and macroscopically visible damage induced by NSAIDs . However , it remains unclear to what extent leukotrienes are involved in the initiation of the injury , as opposed to its amplification .

MH_train_1531

MH_train_1531

MH_train_1531

interacts_with DB00031?

multiple_choice

Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms . A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells , and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase , is described . DB00120 hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity . The phosphorylation site(s) appear to be the same for both kinases . Phosphorylation is accompanied by increased metabolic flux at low , physiologically relevant , substrate concentrations . P01308 and spermine both inhibit the phosphorylation of the enzyme , possibly by increasing dephosphorylation . P17735 is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium . No parallel changes in flux could be detected . Both enzymes are subject to complex regulatory mechanisms , short- and long-term . Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes . Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux . 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 . 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 . Construction , expression and characterization of tissue-type plasminogen activator mutants . Three tissue-type plasminogen activator ( t-PA ) mutants were constructed by recombinant and site-directed mutagenesis techniques . They are del(296-302) with deletion of P05121 binding site , N117Q/N184Q with deglycosylation of P04264 and K2 domains , and their combination mutant designated as GGI . Then these three mutants were successfully transiently expressed in COS-7 cells , and GGI was further stably expressed in CHO cells . The biological characterization of the expression products indicated that del(296-302) and GGI possessed the resistance to inhibition by P05121 . In addition , the specific activity of GGI was increased by about 46 % , the plasma half-life was prolonged by about one fold , while its affinity for fibrin was not affected . 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 . DB00174 synthetase : regulation by cell stress and involvement in tumor biology . DB00174 synthetase ( P08243 ) catalyzes the conversion of aspartate and glutamine to asparagine and glutamate in an DB00171 -dependent reaction . The enzyme is ubiquitous in its organ distribution in mammals , but basal expression is relatively low in tissues other than the exocrine pancreas . Human P08243 activity is highly regulated in response to cell stress , primarily by increased transcription from a single gene located on chromosome 7 . Among the genomic elements that control P08243 transcription is the C/EBP- P39905 response element ( CARE ) within the promoter . Protein limitation or an imbalanced dietary amino acid composition activate the P08243 gene through the amino acid response ( AAR ) , a process that is replicated in cell culture through limitation for any single essential amino acid . Endoplasmic reticulum stress also increases P08243 transcription through the Q9NZJ5 -eIF2- P18848 arm of the unfolded protein response ( UPR ) . Both the AAR and UPR lead to increased synthesis of P18848 , which binds to the CARE and induces P08243 transcription . Elevated expression of P08243 protein is associated with resistance to asparaginase therapy in childhood acute lymphoblastic leukemia and may be a predictive factor in drug sensitivity for certain solid tumors as well . Activation of the Q9P2K8 -eIF2- P18848 signaling pathway , leading to increased P08243 expression appears to be a component of solid tumor adaptation to nutrient deprivation and/or hypoxia . Identifying the roles of P08243 in fetal development , tissue differentiation , and tumor growth may reveal that P08243 function extends beyond asparagine biosynthesis . 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 . 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 . Protein kinase-A activity in P10644 -mutant cells , and regulation of mitogen-activated protein kinases P27361 /2 . Carney complex ( CNC ) is caused by P10644 -inactivating mutations . P10644 encodes the regulatory subunit type I-alpha ( RIalpha ) of the DB02527 -dependent kinase ( PKA ) holoenzyme ; how RIalpha insufficiency leads to tumorigenesis remains unclear . In many cells PKA inhibits the extracellular receptor kinase ( P27361 /2 ) cascade of the mitogen-activated protein kinase ( MAPK ) pathway leading to inhibition of cell proliferation . We investigated whether the PKA-mediated inhibitory effect on P27361 /2 is affected in CNC cells that carry germline P10644 mutations . PKA activity both at baseline and after stimulation with DB02527 was augmented in cells carrying mutations . Quantitative message analysis showed that the main PKA subunits expressed were type I ( RIalpha and RIbeta ) but RIalpha was decreased in mutant cells . Immunoblot assays of P27361 /2 phosphorylation by the cell- and pathway-specific stimulant lysophosphatidic acid ( P08519 ) showed activation of this pathway in a time- and concentration-dependent manner that was prevented by a specific inhibitor . There was a greater rate of growth in mutant cells ; forskolin and isoproterenol inhibited P08519 -induced P27361 /2 phosphorylation in normal but not in mutant cells . DB02587 inhibited P08519 -induced cell proliferation and metabolism in normal cells , but stimulated these parameters in mutant cells . These data were also replicated in a pituitary tumor cell line carrying the most common P10644 mutation ( c.578del TG ) , and an in vitro construct of mutant P10644 that was recently shown to lead to augmented PKA-mediated phosphorylation . We conclude that PKA activity in CNC cells is increased and that its stimulation by forskolin or isoproterenol increases P08519 -induced P27361 /2 phosphorylation , cell metabolism and proliferation . Reversal of PKA-mediated inhibition of this MAPK pathway in CNC cells may contribute to tumorigenesis in this condition . 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 . Overexpression of SnoN/SkiL , amplified at the 3q26.2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26.2 encompassing SnoN/SkiL , a coregulator of SMAD/TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 promoters with little effect on the P38936 promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells . Gremlin gene expression in bovine retinal pericytes exposed to elevated glucose . AIM : To assess the influence of high extracellular glucose on the expression of the bone morphogenetic protein ( BMP ) antagonist , gremlin , in cultured bovine retinal pericytes ( BRPC ) . METHODS : BRPC were cultured under conditions of 5 mM and 30 mM d-glucose for 7 days and total RNA was isolated . Gremlin mRNA levels were correlated , by RT-PCR , with other genes implicated in the pathogenesis of diabetic retinopathy and the signalling pathways in high glucose induced gremlin expression were probed using physiological inhibitors . Gremlin expression was also examined in the retina of streptozotocin induced diabetic mice . RESULTS : High glucose stimulated a striking increase in BRPC gremlin mRNA levels in parallel with increases in mRNA for the growth factors vascular endothelial growth factor ( P15692 ) , transforming growth factor beta ( TGFbeta ) , and connective tissue growth factor ( P29279 ) and changes in other genes including fibronectin and plasminogen activator inhibitor-1 ( P05121 ) . High glucose triggered gremlin expression was modulated by anti-TGFbeta antibody , by the uncoupler of oxidative phosphorylation , CCCP , and by inhibition of Q96HU1 -kinase ( MAPK ) activation . Striking gremlin expression was observed in the outer retina of diabetic mice and also at the level of the vascular wall . CONCLUSIONS : Gremlin gene expression is induced in BRPC in response to elevated glucose and in the retina of the streptozotocin induced diabetic mouse . Its expression is modulated by hyperglycaemic induction of the MAPK , reactive oxygen species , and TGFbeta pathways , all of which are reported to have a role in diabetic fibrotic disease . This implicates a role for gremlin in the pathogenesis of diabetic retinopathy . RNA helicase module in an acetyltransferase that modifies a specific tRNA anticodon . Post-transcriptional RNA modifications in the anticodon of transfer RNAs frequently contribute to the high fidelity of protein synthesis . In eubacteria , two genome-encoded transfer RNA ( tRNA ) species bear the same CAU sequence as the anticodons , which are differentiated by modified cytidines at the wobble positions . The elongator tRNA( DB00134 ) accepts an acetyl moiety at the wobble base to form N(4)-acetylcytidine ( ac(4)C ) : an inherent modification ensures precise decoding of the AUG codon by strengthening C-G base-pair interaction and concurrently preventing misreading of the near cognate AUA codon . We have determined the crystal structure of tRNA( DB00134 ) cytidine acetyltransferase ( TmcA ) from Escherichia coli complexed with two natural ligands , acetyl- DB01992 and ADP , at 2.35 A resolution . The structure unexpectedly reveals an idiosyncratic RNA helicase module fused with a Q92830 -related N-acetyltransferase ( Q969I3 ) fold , which intimately cross-interact . Taken together with the biochemical evidence , we further unravelled the function of acetyl- DB01992 as an enzyme-activating switch , and propose that an RNA helicase motor driven by DB00171 hydrolysis is used to deliver the wobble base to the active centre of the Q969I3 domain . Serial changes in serum vitamin P04264 , triglyceride , cholesterol , osteocalcin and 25-hydroxyvitamin D3 in patients after hip replacement for fractured neck of femur or osteoarthritis . Serum vitamin P04264 concentrations were measured at presentation ( just before surgery ) and then at weekly intervals for 3 weeks in two groups of elderly patients requiring either hemiarthroplasty for fractured neck of femur ( FON , n = 13 ) or total hip replacement for osteoarthritis of the hip ( OA , n = 16 ) . In comparison with healthy elderly volunteers ( n = 25 ) , serum vitamin P04264 concentrations were significantly lower in both groups at presentation , and fell significantly within 24 h after surgery to concentrations approaching non-detectable , subsequently returning to pre-operative values within 3 weeks . Serum vitamin P04264 tended to be lower in the fracture group both before and after operation , although calculation of a vitamin P04264 -triglyceride ratio reduced the apparent difference as triglyceride concentrations were lower in the fracture group . P02818 concentrations were similar and fell significantly after operation in both groups , returning to pre-operative levels within 7 days . No differences in the two forms of osteocalcin ( carboxylated and undercarboxylated ) were observed either before or after operation in either group . DB00146 concentrations were not significantly different between the two groups at any time . DB01022 status may be lower than desirable in certain groups of the elderly population , and supplementation should be considered as prophylactic therapy . [ P01308 resistance and hyperlipidemia in the elderly ] . To elucidate the relationship between hyperlipidemias and insulin resistance in the elderly , we conducted three studies : 1 ) determination of the prevalence of hyperlipidemias in elderly subjects with impaired glucose tolerance or non-insulin dependent diabetes mellitus , 2 ) measurement of plasma glucose and insulin levels in patients with phyerlipidemias and atherosclerotic vascular disease , and 3 ) computation of correlation between levels of substances involved in coagulation and fibrinolysis ( F-VII , F-X , and P05121 ) and levels of triglycerides and insulin in serum in hyperlipidemic patients with atherosclerotic vascular disease . The prevalence of hypertriglyceridemia was 4 % in non-diabetic control subjects , 38 % in subjects with impaired glucose tolerance , 22 % in those with diabetes , and 29 % in those with both conditions . Levels of glucose and insulin in plasma were measured after oral intake of 75 g of glucose . The insulin response was greater in the group with hypertriglyceridemia than in the group with normal triglyceride levels , although the glucose responses did not differ between the groups . The activities and levels of F-VII , F-X , and P05121 correlated with triglycerides in serum and also with fasting insulin levels in hyperlipidemic patients with atherosclerotic vascular disease . We conclude that hypertriglyceridemia plays an important role in the development of atherothrombotic vascular disease ; it accompanies elevation of coagulation and antifibrinolytic activities in elderly patients with insulin resistance . 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 . [ Biological function of fusion protein P39905 -PAI2CD ] . To express the fusion protein P39905 -PAI2CD ( urokinase-type plasminogen activator amino terminal fragment-plasminogen activator inhibitor type 2 with the region inter C and D helices deleted ) gene in E.coli and determine the biological characterization of fusion protein P39905 -PAI2CD , the cDNA fragment encoding P39905 -PAI2CD was cloned into the expression vector pLY-4 and transformed into E.coli JF1125 . After temperature induction , the expression amount of P39905 -PAI2CD account for 15 % of total bacterial protein . The result was confirmed by Western blot . P39905 -PAI2CD protein was isolated and purified by washing and solubilization of inclusion body , renaturation and ion exchange chromatography . The final product displayed a single band with a corresponding molecular weight 62 kD in SDS-PAGE . The purity was over 90 % , the protein yield was 50 % and the specific activity was 12 000 IU/mg . The P05121 activity was measured by chromogenic assay . The purified fusion protein inhibited urokinase-type plasminogen activator as measured by milk-agarose plate assay , and bound to human lung cancer cells via uPA receptor ( Q03405 ) , which was confirmed by radio competition experiments . The results indicate that the biological characteristics of P39905 -PAI2CD were very similar to those of the wide type P05120 ( or mutants P05120 , P05121 -2CD ) and to pro-uPA in binding to Q03405 -bearing cells . DB01270 for age-related macular degeneration . INTRODUCTION : Age-related macular degeneration ( AMD ) is the leading cause of blindness in patients over 50 years in the developed world . The wet form of AMD is responsible for the majority of severe vision loss . P15692 is a key component in the development of wet AMD . DB01270 is an anti- P15692 agent that has established itself as the gold standard in the treatment of neovascular AMD . Herein , we review the pharmacology , pharmacokinetics , efficacy and safety of ranibizumab . AREAS COVERED : Since its approval in 2006 , ranibizumab has revolutionized the treatment of wet AMD . In two pivotal Phase III trials , MARINA and ANCHOR , ranibizumab ( 0.5 mg ) prevented moderate visual loss in 90 and 96 % of patients , respectively , and improved vision by 15 letters or more in 33 and 40 % of patients , respectively . Fixed monthly dosing regimens were compared with quarterly dosing regimens in PIER and EXCITE studies and support the superiority of fixed monthly dosing . The CATT trial revealed that bevacizumab was not inferior to ranibizumab when dosed monthly . As-needed treatment regimens of ranibizumab were also found to be non-inferior to monthly ranibizumab after 1 year of follow-up . EXPERT OPINION : DB01270 has positively altered the treatment of wet AMD and offers hope for millions of patients . Extracellular signal-regulated kinase and the small GTP-binding protein , Rac , contribute to the effects of transforming growth factor-beta1 on gene expression . The kinases and regulatory proteins that convey signals initiated by transforming growth factor-beta ( TGF-beta ) to the nucleus are poorly characterized . To study the role of the extracellular signal-regulated kinase ( P29323 ) pathway in this process , we transiently transfected NIH 3T3 fibroblasts with TGF-beta-responsive luciferase reporter genes and expression vectors designed to interrupt this kinase cascade . Mitogen-activated protein ( Q96HU1 ) kinase phosphatase-1 and a dominant negative Q96HU1 / P29323 kinase 1 mutant reduced stimulation of plasminogen activator inhibitor-1 ( P05121 ) promoter activity by TGF-beta1 from 11.5- to 4-fold and 4.9-fold , respectively . Similar results were observed with the type I collagen promoters . TGF-beta1 increased P27361 activity 4.5-fold at 5 min and 3 . 1-fold at 3 h , while Jun kinase and p38 activity were not affected . Cotransfection of a dominant negative mutant of the small G protein , Rac , but not dominant negative Ras , Cdc42 , or Rho mutants , reduced the effects of TGF-beta1 on the P05121 promoter by approximately half . In support of a role for Rac in signaling by TGF-beta , GTP binding to Rac was increased 3.7-fold following exposure of NIH 3T3 cells to TGF-beta1 for 3 min . These findings indicate that TGF-beta1 modulates gene expression partly through P29323 and Rac in NIH 3T3 cells . The histone acetyltransferase Q92831 regulates P38936 transcription through stress-induced acetylation of histone H3 . The activity of p53 as a tumor suppressor primarily depends on its ability to transactivate specific target genes in response to genotoxic and other potentially mutagenic stresses . Several histone acetyl transferases ( HATs ) , including p300 , CBP , Q92831 and Q92830 have been implicated in the activation of p53-dependent transcription of the cyclin-dependent kinase ( cdk ) inhibitor P38936 as well as other target genes . Here we show that Q92831 , but not CBP or p300 , is a critical regulator of p53-dependent P38936 expression in response to multiple p53-activating stresses . Q92831 was required for the transcriptional activation of P38936 in response to exogenous p53 in p53-null cells , nutlin-3 , DNA damaging agents and p14( Q8N726 ) expression , suggesting a broad requirement for Q92831 in p53 signaling to P38936 after stress . Importantly , cells lacking Q92831 failed to undergo cell cycle arrest in response to nutlin-3 treatment or p14( Q8N726 ) expression , consistent with a physiologically important role for Q92831 in this p53 function . Surprisingly , the role for Q92831 in induction of P38936 was independent of p53 lysine 320 acetylation , a previously suggested target of Q92831 -mediated acetylation . Though P38936 promoter occupancy by p53 was not altered by Q92831 knockdown , activation of P38936 transcription required an intact Q92831 O60235 domain , and induction of chromatin marks acetyl-H3K9 and acetyl-H3K14 at the P38936 promoter by p53 was dependent upon physiologic levels of Q92831 . Together , our experiments indicate that Q92831 is required for stress-responsive histone 3 acetylation at the P38936 promoter , p53-directed transcription of P38936 and the resultant growth arrest .

MH_train_1532

MH_train_1532

MH_train_1532

interacts_with DB00682?

multiple_choice

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 . 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 . Polymorphisms of the HTR1a allele are linked to frontal brain electrical asymmetry . Polymorphic variations in genes related to serotonin synthesis , transport , recognition , or degradation may convey subtle changes in serotonin system architecture that may place an individual at risk for psychopathology when faced with life stressors . The relationship between three key serotonin alleles and frontal brain electrical asymmetry , a putative endophenotype of depression , was examined . Risk alleles were hypothesized to predict relatively greater right frontal brain activity regardless of current clinical state . A sample of 313 college-age individuals , spanning a range of depressive severity from no symptomotology to clinically meaningful levels , participated . Resting encephalographic ( EEG ) activity was recorded from 64 scalp sites on four occasions separated by at least 24h ( two 8-min recording sessions occurring at each occasion ) . Alpha power asymmetry scores between homologous sites were calculated for each session and then averaged to form a trait metric of asymmetry for each pair . PCR based genotyping was conducted for the HTR1a , HTR2a , and HTTLPR genes . Variations in the HTR1a gene were related to trait EEG asymmetry , regardless of any history of depression . Compared to subjects with at least one non-risk allele , subjects with homozygous P08908 risk alleles had significantly greater relative right frontal activity at sites P08709 / P00451 , P12259 /F6 , and F1/F2 . In conclusion , variation in HTR1a can influence trait level brain activity , which may ultimately be indicative of risk for psychopathology . 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 . 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 . 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 . DB00114 values in cerebrospinal fluid : reference values and diagnosis of Q9NVS9 deficiency in paediatric patients . Our aim was to establish reference values for cerebrospinal fluid ( P04141 ) pyridoxal 5'-phosphate ( PLP ) in a paediatric population for the diagnosis of pyridox(am)ine 5'-phosphate oxidase ( Q9NVS9 ) deficiency . For reference values , P04141 samples from 113 paediatric controls ( age range : 1 day-18 years ) from Barcelona and London were analysed . Cerebrospinal fluid PLP and biogenic amine concentrations were analysed by HPLC with fluorescence and electrochemical detection . DB00114 concentrations in 4 patients with Q9NVS9 deficiency were determined . A negative correlation between P04141 PLP values and age of controls was observed in both populations ( r=-0.503 ; p < 0.0001 and r=-0.542 ; p=0.002 ) . Reference values were stratified into 4 ( Barcelona ) and 3 age groups ( London ) . For the newborn period , P04141 PLP reference intervals were 32-78 and 44-89 nmol/L for the Barcelona and London centers , respectively ) . No correlation was observed in the different age groups between PLP values and biogenic amines metabolites . PLP values in neonates with Q9NVS9 deficiency were clearly decreased ( PLP=3.6 , 12.0 , 14.0 and 18.0 nmol/L ) compared with our reference ranges . In conclusion , reference values for P04141 PLP should be stratified according to age . No association was observed between PLP values and biogenic amines metabolites . In our 4 cases with Q9NVS9 deficiency , P04141 PLP values were clearly below the reference values . The protective effect of rebamipide on paracellular permeability of rat gastric epithelial cells . BACKGROUND : Barrier function in gastric epithelial cells is essential for the gastric defence mechanism against acid back-diffusion into the mucosal layer . Our previous study indicated that trans-epithelial resistance ( Q9NZ01 ) of rat gastric epithelial cells was rapidly increased when the cells were exposed to acid . This response to acid was diminished by indometacin . AIM : Evaluate the effects of a mucoprotective agent , rebamipide , on the nonsteroidal anti-inflammatory drug ( NSAID ) -induced increase of gastric epithelial permeability . METHODS : Rat gastric epithelial cells were plated on tissue culture inserts . Cells were exposed to a NSAID ( indometacin , 10-7 M ) . Trans-epithelial permeability was measured by Q9NZ01 and diffusion rate of 14C-mannitol . The effect of rebamipide was evaluated by measuring Q9NZ01 . Endogenous prostaglandin E2 ( DB00917 ) production in culture medium was also measured . RESULTS : DB00328 gradually and significantly decreased Q9NZ01 and increased 14C-manitol permeability . Rebamipide reversed the indometacin-induced changes in epithelial permeability and induced DB00917 synthesis . This induction was blocked by either indometacin or a Cyclooxygenase ( P36551 ) -2 specific inhibitor . CONCLUSIONS : P36551 inhibitors such as indometacin inhibit regulation of epithelial permeability by reducing DB00917 . P23219 has an important role in the gastric defense mechanism . Rebamipide suppressed an indometacin-induced increase in gastric epithelial permeability by increasing DB00917 levels in a P35354 dependent manner . 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 . [ Association of allelic polymorphisms of genes matrix Gla-protein system with ischemic atherothrombotic stroke ] . There are results of the determination of 10 polymorphisms of matrix Gla-protein system ( gene P08493 -T(-138) --> C ( rs1800802 ) , G(-7) --> A ( rs1800801 ) , Thr83 --> Ala ( rs4236 ) , gene P11473 -FokI ( rs2228570 ) , BsmI ( rs1544410 ) , ApaI ( rs7975232 ) , TaqI ( rs731236 ) , gene P38435 -Arg325 --> Gln ( rs699664 ) , gene VKORS1-T(2255) --> C ( rs2359612 ) , gene P12643 -Ser37 --> Ala ( rs2273073 ) ) into 170 patients with ischemic atherothrombotic stroke ( IATS ) and 124 healthy individual is ( control group ) . It is established that there is a connection between the IATS and polymorphic variants of genes P08493 ( G(-7) --> A ) and Q9BQB6 ( T(2255) --> C ) . The risk of IATS in carriers of minor allele A/A ( G(-7) --> A polymorphism ) in 2.6 times higher than in carriers of the major allele ( G/A + G/G ) , and C/C genotype ( T(2255) --> C polymorphism ) in 2.2 times higher than the homozygotes of major allele . The coincidence of patients T/C and G/G , C/C and G/A genotypes , and A/A genotype ( G(-7) --> A polymorphism ) with any genotype T(2255) --> C polymorphism are increases the risk of IATS . 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 . 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 . Whole-exome sequencing and imaging genetics identify functional variants for rate of change in hippocampal volume in mild cognitive impairment . Whole-exome sequencing of individuals with mild cognitive impairment , combined with genotype imputation , was used to identify coding variants other than the apolipoprotein E ( P02649 ) ε4 allele associated with rate of hippocampal volume loss using an extreme trait design . Matched unrelated P02649 ε3 homozygous male Caucasian participants from the Alzheimer 's Disease Neuroimaging Initiative ( ADNI ) were selected at the extremes of the 2-year longitudinal change distribution of hippocampal volume ( eight subjects with rapid rates of atrophy and eight with slow/stable rates of atrophy ) . We identified 57 non-synonymous single nucleotide variants ( SNVs ) which were found exclusively in at least 4 of 8 subjects in the rapid atrophy group , but not in any of the 8 subjects in the slow atrophy group . Among these SNVs , the variants that accounted for the greatest group difference and were predicted in silico as ' probably damaging ' missense variants were rs9610775 ( Q9BWT7 ) and rs1136410 ( P09874 ) . To further investigate and extend the exome findings in a larger sample , we conducted quantitative trait analysis including whole-brain search in the remaining ADNI P02649 ε3/ε3 group ( N=315 ) . Genetic variation within P09874 and Q9BWT7 was associated with rate of hippocampal neurodegeneration in P02649 ε3/ε3 . Meta-analysis across five independent cross sectional cohorts indicated that rs1136410 is also significantly associated with hippocampal volume in P02649 ε3/ε3 individuals ( N=923 ) . Larger sequencing studies and longitudinal follow-up are needed for confirmation . The combination of next-generation sequencing and quantitative imaging phenotypes holds significant promise for discovery of variants involved in neurodegeneration . 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 . Molecular basis for the immunostimulatory activity of guanine nucleoside analogs : activation of Q9NYK1 . Certain Q99618 -substituted and N7 , Q99618 -disubstituted guanine ribonucleosides comprise a class of small molecules with immunostimulatory activity . In a variety of animal models , these agents stimulate both humoral and cellular immune responses . The antiviral actions of these guanosine analogs have been attributed to their ability to induce type I IFNs . However , the molecular mechanisms by which the guanosine analogs potentiate immune responses are not known . Here , we report that several guanosine analogs activate Q9NYK1 ( Q9NYK1 ) . DB04860 , 7-deazaguanosine , and related guanosine analogs activated mouse immune cells in a manner analogous to known TLR ligands , inducing cytokine production in mouse splenocytes ( P05231 and IL-12 , type I and II IFNs ) , bone marrow-derived macrophages ( P05231 and IL-12 ) , and in human peripheral blood leukocytes ( type I IFNs , tumor necrosis factor alpha and IL-12 ) . The guanosine congeners also up-regulated costimulatory molecules and MHC I/II in dendritic cells . Genetic complementation studies in human embryonic kidney 293 cells confirmed that the guanosine analogs activate cells exclusively via Q9NYK1 . The stimulation of Q9NYK1 by the guanosine analogs in human cells appears to require endosomal maturation because inhibition of this process with chloroquine significantly reduced the downstream activation of NF-kappaB . However , Q9NR97 activation by R-848 and O60603 activation by [ S-[2,3-bis(palmitoyloxy)-(2-RS)-propyl]-N-palmitoyl-R- DB00151 -S- DB00133 -Lys4-OH , trihydrochloride ) ] were not inhibited by chloroquine , whereas Q9NR96 activation by CpG oligodeoxynucleotides was abolished . In summary , we present evidence that guanosine analogs activate immune cells via Q9NYK1 by a pathway that requires endosomal maturation . Thus , the B cell-stimulating and antiviral activities of the guanosine analogs may be explained by their Q9NYK1 -activating capacity . Structure functional expression and spatial distribution of a cloned cDNA encoding a rat P28221 -like receptor . Using polymerase chain reaction ( PCR ) a complementary DNA ( cDNA ) encoding a 5-hydroxytryptamine ( 5-HT ) receptor was isolated from rat forebrain . The amplified cDNA specifies an open reading frame of 374 amino acids comprising seven putative transmembrane regions . Expression of the cloned cDNA in human embryonic kidney cells ( P29320 293 ) was used to establish the pharmacological profile of the encoded receptor polypeptide . Membranes containing the cloned receptor showed high affinity binding of [ 3H ] -5-HT . Competition binding experiments with a variety of serotonin receptor ligands displayed a rank order of affinities corresponding to a P28221 subtype : 5-CT > 5-HT = metergoline > CGS 12066 > methysergide > sumatriptan > mianserin = (-)alpha-Me-5-HT = yohimbine > 8-OH-DPAT > or = rauwolscine > spiperone > DOI > propranolol > or = 2-Me-5-HT > or = ICS 205930 . Ketanserin and ritanserin displaced [ 3H ] -5-HT-binding in a biphasic manner . In situ hybridization revealed highest expression of the corresponding mRNA in the pyramidal layer of the olfactory tubercle and the nucleus caudatus and accumbens . 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 . Randomised clinical trial : effects of monotherapy with DB05070 , a P41594 inhibitor , on symptoms and reflux events in patients with gastro-oesophageal reflux disease . BACKGROUND : DB05070 , a metabotropic glutamate receptor 5 ( P41594 ) negative allosteric modulator , has been shown to reduce gastro-oesophageal reflux events and oesophageal acid exposure in patients with gastro-oesophageal reflux disease ( GERD ) and healthy subjects . AIM : To evaluate the effects of DB05070 monotherapy for 2 weeks on symptom control in patients with GERD . METHODS : This was a double-blind , placebo-controlled , multi-centre trial in GERD patients who were responders to proton pump inhibitors ( PPIs ) . Following PPIs withdrawal , a 2-week baseline washout period was followed by 2-week treatment with either DB05070 120 mg or placebo b.d . The primary clinical efficacy endpoint was the number of GERD symptom-free days in treatment week 2 compared with the last 7 days of baseline . The effect on reflux events using 24-h impedance-pH monitoring was also determined in a subset of 24 patients . RESULTS : The full analysis set comprised 103 patients DB05070 ( N= 50 ) , Placebo ( N=53 ) . In treatment week 2 , DB05070 significantly increased GERD symptom-free days ( P=0.045 ) and heartburn-free days ( P=0.037 ) , reduced antacid use ( P=0.017 ) , improved total symptom score ( P=0.048 ) including subscale heartburn/regurgitation ( P=0.007 ) and sleep disturbance because of GERD ( P= 0.022 ) . DB05070 significantly reduced total ( P=0.034 ) and acidic reflux events ( P=0.003 ) . DB05070 was well tolerated . Most common adverse events for DB05070 were mild to moderate dizziness 16 % and vertigo 12 % ( placebo 4 % and 2 % ) . CONCLUSIONS : Inhibition of P41594 with DB05070 monotherapy reduces reflux events and improves symptoms in GERD patients . This mechanism has promise for the management of GERD . Substituent effects of N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamides on positive allosteric modulation of the metabotropic glutamate-5 receptor in rat cortical astrocytes . CDPPB [ 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide ] was recently described as the first centrally active , positive allosteric modulator of rat and human metabotropic glutamate receptor ( mGluR ) P41594 subtype . We explored the structural requirements for potentiation of glutamate-induced calcium release in naturally expressed P41594 in cultured rat astrocytes and increasing affinity for the allosteric antagonist binding site by evaluating 50 analogues of CDPPB . In the fluorometric calcium assay , CDPPB exhibited an EC50 value of 77 +/- 15 nM in potentiating P41594 -mediated responses in cortical astrocytes and a Ki value of 3760 +/- 430 nM in displacing [3H]methoxyPEPy binding in membranes of cultured P29320 -293 cells expressing rat P41594 . The structure-activity relationships showed that electronegative aromatic substituents in the para-position of the benzamide moiety of CDPPB increase potency . Both binding and functional activities were further increased with a halogen atom in the ortho-position of the 1-phenyl ring . These effects of substitution do not match those of either aromatic ring of MPEP [ 2-methyl-6-(phenylethynyl)pyridine ] for the antagonist allosteric binding site . Combination of the optimal substituents and aromatic positions resulted in 4-nitro-N-(1-(2-fluorophenyl)-3-phenyl-1H-pyrazol-5-yl)benzamide ( VU-1545 ) showing Ki = 156 +/- 29 nM and EC50 = 9.6 +/- 1.9 nM in the binding and functional assays , respectively . Novel targeted therapeutics : inhibitors of Q00987 , Q9UM73 and PARP . We reviewed preclinical data and clinical development of Q00987 ( murine double minute 2 ) , Q9UM73 ( anaplastic lymphoma kinase ) and PARP ( poly [ ADP-ribose ] polymerase ) inhibitors . Q00987 binds to p53 , and promotes degradation of p53 through ubiquitin-proteasome degradation . JNJ-26854165 and RO5045337 are 2 small-molecule inhibitors of Q00987 in clinical development . Q9UM73 is a transmembrane protein and a member of the insulin receptor tyrosine kinases . Q9HC35 - Q9UM73 fusion gene is identified in approximately 3-13 % of non-small cell lung cancer ( NSCLC ) . Early-phase clinical studies with DB08865 , an Q9UM73 inhibitor , in NSCLC harboring Q9HC35 - Q9UM73 have demonstrated promising activity with high response rate and prolonged progression-free survival . PARPs are a family of nuclear enzymes that regulates the repair of DNA single-strand breaks through the base excision repair pathway . Randomized phase II study has shown adding P09874 inhibitor BSI-201 to cytotoxic chemotherapy improves clinical outcome in patients with triple-negative breast cancer . DB09074 , another oral small-molecule PARP inhibitor , demonstrated encouraging single-agent activity in patients with advanced breast or ovarian cancer . There are 5 other PARP inhibitors currently under active clinical investigation . 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 . Evidence for serotonin (5-HT)1B , P28221 and P30939 receptor inhibitory effects on trigeminal neurons with craniovascular input . Development of serotonin ( 5HT(1B/1D) ) agonists for the acute attack of migraine resulted in considerable interest in their action . The superior sagittal sinus ( SSS ) was isolated in alpha-chloralose ( 60 mg/kg , i.p. and 20 mg/kg i.v.i. supplementary 2 hourly ) anaesthetised cats . The SSS was stimulated electrically ( 100 V , 250 micros duration , 0.3 Hz ) and neurons of the trigeminocervical complex monitored using electrophysiological methods . To test 5-HT(1B) receptor-mediated activity common carotid blood flow ( CCF ) was monitored with a transonic flow probe placed around the vessel . DB00952 ( 5-HT(1B/1D/1F) receptor agonist ) and alniditan ( 5-HT(1B/1D) receptor agonist ) produced reductions in carotid blood flow of 38+/-5 % and 42+/-6 % , respectively . These effects were attenuated by the 5-HT(1B) receptor antagonist SB224289 ( P < 0.05 ) . LY344864 ( 5-HT(1F) receptor agonist ) had no effect on CCF . DB00952 inhibited SSS-evoked activity ( 61+/-7 % ) , an effect partially inhibited by the 5-HT(1B) receptor antagonist SB224289 ( 30+/-5 % ) , or by the 5-HT(1D) receptor antagonist O95696 -15572 ( 37+/-6 % ) . There remained an inhibitory effect of naratriptan after both 5-HT(1B) and 5-HT(1D) receptor blockade ( 22+/-5 % ) . Alniditan inhibited SSS-evoked trigeminal activity ( 53+/-6 % ) , an effect abolished after 5-HT(1B) and 5-HT(1D) receptor blockade . LY344864 ( 5-HT(1F) receptor agonist ) inhibited SSS-evoked trigeminal activity ( 28+/-5 % ) , an effect unaltered by either SB224289 or O95696 -15572 . It can be concluded that there are inhibitory 5-HT(1B) , 5-HT(1D) and 5-HT(1F) receptors in the trigeminocervical complex of the cat . 5-HT(1B) receptor-mediated inhibition is the most potent of the three in terms of inhibition of trigeminovascular nociceptive traffic . [ Possible application of pharmacogenomics to warfarin therapy ] . Vitamin K-dependent gamma-carboxylation system is crucial to generate active coagulation factors . It consists of gamma-Glutamylcarboxylase ( P38435 ) and vitamin K-epoxide reductase ( Q9BQB6 ) . DB00682 is an anticoagulant that blocks Q9BQB6 . Recent studies have shown that genetic variations in a subunit of Q9BQB6 complex , Q9BQB6 , and in cytochrome P 450 ( CYP ) 2C9 genes are strong determinants of individuals ' warfarin sensitivity . Algorithms have been proposed to predict warfarin doses , and about 55 % of the variance in warfarin dose could be attributed to variations in the Q9BQB6 and P11712 genes together with age , sex , body-surface area , and presence or absence of heart valve replacement . Contributions of polymorphisms in Q9BQB6 and P11712 to inter-individual differences of warfarin dose , however , are different among races . Studies have shown that potential clinical and economic benefits of P11712 / Q9BQB6 genotype-guided dosing are only marginal . Thus , evidence is still limited and application of warfarin pharmacogenomics to clinical practice at present needs careful consideration .

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interacts_with DB00227?

multiple_choice

Q96RI1 , a bile acid receptor and biological sensor . Bile acid synthesis is a major pathway for cholesterol disposal and thus represents a potential therapeutic target pathway for the treatment of hypercholesterolemia . Recently , the nuclear farnesoid X receptor ( Q96RI1 ) was identified as a bile acid receptor and biological sensor for the regulation of bile acid biosynthesis . Q96RI1 was shown to regulate cholesterol metabolism in two ways : ( 1 ) chenodeoxycholic acid ( DB06777 ) , a primary bile acid , binds directly to and activates Q96RI1 , which then mediates the feedback suppression by bile acids of cholesterol 7 alpha-hydroxylase ( P22680 ) , the rate-limiting enzyme in bile acid biosynthesis from cholesterol ; and ( 2 ) Q96RI1 participates in the activation of intestinal bile acid binding protein ( IBABP ) , which is involved in the enterohepatic circulation of bile acids . Thus Q96RI1 constitutes a potential therapeutic target that can be modulated to enhance the removal of cholesterol from the body . DB08875 suppresses tumor growth and metastasis in hepatocellular carcinoma by a dual blockade of P35968 and MET . PURPOSE : MET signaling has been suggested a potential role in hepatocellular carcinoma ( HCC ) and associated with prometastasis during antiangiogenesis therapy . We investigated the potential association between MET expression and therapeutic response to sorafenib in patients with HCC . Antitumor effects of cabozantinib , a dual inhibitor of MET and P35968 , were examined in cultured HCC cells as well as in vivo models . EXPERIMENTAL DESIGN : Total MET and phosphorylated MET ( p-MET ) were measured in 29 resected HCC specimens , and correlated with response to sorafenib as postoperative adjuvant therapy . In the second set of experiments using cultured HCC cells , and mouse xenograft and metastatic models , effects of cabozantinib were examined . RESULTS : High level of p-MET in resected HCC specimens was associated with resistance to adjuvant sorafenib therapy . In cultured HCC cells that expressed p-MET , cabozantinib inhibited the activity of MET and its downstream effectors , leading to P55008 -phase arrest . DB08875 inhibited tumor growth in p-MET-positive and p-MET-negative HCC by decreasing angiogenesis , inhibiting proliferation , and promoting apoptosis , but it exhibited more profound efficacy in p-MET-positive HCC xenografts . DB08875 blocked the hepatocyte growth factor ( P14210 ) -stimulated MET pathway and inhibited the migration and invasion of the HCC cells . Notably , cabozantinib reduced the number of metastatic lesions in the lung and liver in the experimental metastatic mouse model . CONCLUSIONS : Patients with HCC with high level of p-MET are associated with resistance to adjuvant sorafenib treatment . The dual blockade of P35968 and MET by cabozantinib has significant antitumor activities in HCC , and the activation of MET in HCC may be a promising efficacy-predicting biomarker . Clin Cancer Res ; 20(11) ; 2959-70 . ©2014 AACR . Characterization of a delta opioid receptor in rat pheochromocytoma cells . In one subclone of PC12 pheochromocytoma cells , PC12h , the levels of delta opioid receptors markedly increase in response to nerve growth factor ( P01138 ) . This increase , as assessed by [3H]diprenorphine binding , is found only under specific culture conditions . P01138 treatment of PC12h cells also results in the induction of delta opioid receptor ( P41143 ) mRNA . The time course for P01138 induction of mRNA and protein is similar , although the levels of mRNA increase approximately 5-fold , whereas the levels of receptor increase only 2-fold . Competition studies with selective delta ( [ D-Pen2,5 ] -enkephalin , DPDPE ) , mu ( [ D-Ala2,N-Me-Phe4, DB00145 -ol5 ] - enkephalin , DAMGO ) and kappa ( trans-[+/-]-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]- benzenacetamide methansulfonate salt , U50,488 ) opioid agonists to displace [3H]diprenorphine confirm that the delta subtype of opioid receptor is present on PC12h cells . The delta opioid receptor is coupled functionally , as indicated by agonist inhibition of forskolin-stimulated DB02527 accumulation in a naloxone-reversible manner . Finally , the delta opioid receptor was cloned from PC12h cells . The sequence was identical with that described previously for the rat clone . The PC12h cell line thus provides a model system in which to study regulation of delta opioid receptors . DB00227 . A preliminary review of its pharmacodynamic properties and therapeutic use in hyperlipidaemia . DB00227 is the first of a new class of cholesterol lowering drugs that competitively inhibit P04035 . This new drug decreases cholesterol synthesis and apolipoprotein B concentrations , and increases P01130 activity without adverse effects on other products in the cholesterol pathway . In patients with heterozygous familial or polygenic ( non-familial ) hypercholesterolaemia , oral lovastatin 20 to 40 mg twice daily reduces plasma total cholesterol and LDL-cholesterol concentrations by 25 to 40 % over a period of several weeks . DB00227 also produces decreases in plasma triglyceride and VLDL-cholesterol concentrations , although to a lesser extent . In addition , small though significant increases in HDL-cholesterol concentrations have been observed . Combined administration of lovastatin with other lipid-lowering drugs results in further reductions in plasma total and LDL-cholesterol concentrations beyond those seen with either drug alone . From findings in short term studies , lovastatin appears to be well tolerated with a low incidence of side effects . However , liver function tests and eye examinations for possible lens opacities are advised , and further long term studies in larger groups of patients are necessary before the side effect profile of lovastatin will be clearly established . As would be expected at this relatively early stage of its clinical ' life , ' lovastatin has not yet been studied in a manner that would determine its effect on cardiovascular mortality during long term administration . Nevertheless , if the substantial improvements to patients ' lipid and lipoprotein profiles observed in short term studies are maintained during long term administration , then lovastatin will have an important role in the pharmacological management of hyperlipidaemia . Phosphorylation at serine 208 of the 1alpha,25-dihydroxy P11473 modulates the interaction with transcriptional coactivators . Upon ligand binding the 1alpha,25-dihydroxy P11473 ( P11473 ) undergoes a conformational change that allows interaction with coactivator proteins including P52701 / P12931 family members and the multimeric DRIP complex through the Q15648 subunit . Casein kinase II ( CKII ) phosphorylates P11473 both in vitro and in vivo at serine 208 within the hinge domain . This phosphorylation does not affect the ability of P11473 to bind DNA , but increases its ability to transactivate target promoters . Here , we have analyzed whether phosphorylation of P11473 by CKII modulates the ability of P11473 to interact with coactivators in vitro . We find that both mutation of serine 208 to aspartic acid ( VDRS208D ) or phosphorylation of P11473 by CKII enhance the interaction of P11473 with Q15648 in the presence of 1alpha,25-dihydroxy DB00169 . We also find that the mutation VDRS208D neither affects the ability of this protein to bind DNA nor to interact with Q15788 and RXRalpha . Together , our results indicate that phosphorylation of P11473 at serine 208 contributes to modulate the affinity of P11473 for the DRIP complex and therefore may have a role in vivo regulating P11473 -mediated transcriptional enhancement . Atomic force microscopy study of the effect of HER 2 antibody on P01133 mediated ErbB ligand-receptor interaction . P04626 , a member of the epidermal growth factor receptor ( ErbB ) family , is over-expressed in many cancers . DB00072 and DB06366 are two monoclonal antibodies targeting different extracellular domains of P04626 for cancer therapy . As DB06366 binds to the dimerization arm of P04626 , it can block P04626 heterodimerization and in turn ErbB signaling . Whether DB00072 has the same function is unclear . In this work , we have applied living-cell single-molecule force spectroscopy ( SMFS ) by Atomic Force Microscopy ( P43652 ) to investigate the effect of DB00072 , as well as DB06366 , on P04626 -modulated P01133 - P00533 interaction . The results demonstrated that P01133 bound to P00533 more stably in the cells co-expressing P00533 and P04626 , and the binding enhancement in the presence of P04626 was inhibited by either DB00072 or DB06366 . DB00072 is expected to exert a similar inhibition effect on P04626 / P00533 dimerization as DB06366 , although it does not bind directly to the dimerization arm of P04626 . FROM THE CLINICAL EDITOR : Living-cell single-molecule force spectroscopy ( SMFS ) combined by Atomic Force Microscopy ( P43652 ) was used by this team of scientists to investigate the effect of two monoclonal antibodies used in cancer therapy , DB00072 and DB06366 , on P04626 -modulated P01133 - P00533 interaction , demonstrating the utility of this technique in characterizing the effects of protein-based therapeutics on membrane receptors . 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 . 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 . 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 . A pilot pharmacologic biomarker study of busulfan and fludarabine in hematopoietic cell transplant recipients . PURPOSE : Sixteen patients diagnosed with various hematologic malignancies participated in a phase II study evaluating the addition of rabbit antithymocyte globulin ( DB00098 , DB00098 (®) ) to the hematopoietic cell transplant ( HCT ) conditioning regimen of IV fludarabine monophosphate ( fludarabine ) and targeted intravenous ( IV ) busulfan ( fludarabine/(T)busulfan ) . Our goal was to evaluate pharmacologic biomarkers pertinent to both medications in these patients . METHODS : We characterized the interpatient variability of pharmacologic biomarkers relevant to busulfan , specifically busulfan concentration at steady state , and fludarabine , specifically F-ara-A area under the curve ( AUC ) and fludarabine triphosphate ( F-ara- DB00171 ) intracellular accumulation and concentration in separate P01730 (+) and CD8(+) T-lymphocyte populations . RESULTS : Acute and chronic graft versus host disease ( GvHD ) occurred in 11 patients and one patient , respectively . Four patients died before day +100 of non-relapse causes , which met the protocol stopping guidelines . The cumulative incidence of relapse was 25 % at 3 year post-HCT . Interpatient variability in the busulfan- and fludarabine-relevant pharmacologic biomarkers was 2.1- to 2.5-fold . F-ara-A AUC and accumulated F-ara- DB00171 in CD8(+) cells had the highest hazard ratio for non-relapse mortality and overall survival , respectively . However , neither achieved statistical significance . CONCLUSIONS : The low rates of GvHD , particularly in its chronic form , were encouraging , and further biomarker studies are warranted to optimize the fludarabine/(T)busulfan/ DB00098 conditioning regimen . The role of the sympathetic efferents in endotoxin-induced localized inflammatory hyperalgesia and cytokine upregulation . The sympathetic system ( SNS ) is considered to be a major component of the neurogenic contribution to inflammation and hyperalgesia . We have investigated the role of the SNS in the local inflammatory pain induced by intraplantar ( i.pl ) injections of bacterial endotoxin ( ET ) . Treatment of rats with an alpha-adrenoceptor antagonist ( phentolamine , 0.25-1 mg/kg , i.p. ) , a beta-adrenoceptor antagonist ( propranolol , 1-10 mg/kg , p.o. ) or a sympathetic neuron-blocking agent ( guanethedine , 30 mg/kg , s.c. ) resulted in a dose-dependent reduction of the thermal hyperalgesia induced by ET . Mechanical hyperalgesia , however , was less sensitive to inhibition by propranolol and guanethedine but significantly inhibited by phentolamine . ET injection produced significant upregulation of tumor necrosis factor-alpha ( P01375 ) , interleukin-1 beta ( P01584 ) , P05231 , and nerve growth factor ( P01138 ) . Treatment with any one of the three sympatholytics abolished the upregulation of P01138 and P05231 , while phentolamine and guanethedine also reversed the upregulation of P01375 . P01584 was resistant to all of the sympatholytic treatments . We conclude that the SNS can contribute to the local inflammation and hyperalgesia following injection of ET . The resistance to sympatholytics shown by P01584 , known to play a key role in the inflammatory cascade , suggests that ET can initiate inflammation and hyperalgesia independently of peripheral and central sympathetic mechanisms . DB00227 overcomes gefitinib resistance in human non-small cell lung cancer cells with K-Ras mutations . DB00227 is a 3-hydroxy-3-methylglutaryl-coenzyme A ( HMG- DB01992 ) reductase inhibitor . Its inhibitory action on P04035 leads to depletion of isoprenoids , which inhibits post-translational modification of DB01367 . In this study , we investigated the effect of combining lovastatin with gefitinib on gefitinib-resistant human non-small cell lung cancer ( NSCLC ) cell lines with K-Ras mutations . Antitumor effects were measured by growth inhibition and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay . Effects on apoptosis were determined by flow cytometry , DNA fragmentation , and immunoblots . Protein levels of DB01367 , AKT/pAKT , and RAF/ P27361 /2 in cancer cells were analyzed by immunoblot . Compared with gefitinib alone , a combination of gefitinib with lovastatin showed significantly enhanced cell growth inhibition and cytotoxicity in gefitinib-resistant A549 and NCI-H460 human NSCLC cells . In addition , lovastatin combination treatment significantly increased gefitinib-related apoptosis , as determined by fluorescence microscopy and flow cytometric analysis . These effects correlated with up-regulation of cleaved caspase-3 , poly ( ADP-ribose ) polymerase ( PARP ) , and Bax and down-regulation of Bcl-2 . The combination of lovastatin and gefitinib effectively down-regulated DB01367 protein and suppressed the phosphorylation of RAF , P27361 /2 , AKT , and P00533 in both cell lines . Taken together , these results suggest lovastatin can overcome gefitinib resistance , in NSCLC cells with K-Ras mutations , by down regulation of DB01367 protein , which leads to inhibition of both RAF/ P29323 and AKT pathways . 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 . Molecular pathways : targeting mechanisms of asbestos and erionite carcinogenesis in mesothelioma . Malignant mesothelioma is an aggressive malignancy related to asbestos and erionite exposure . AP-1 transcriptional activity and the NF-κB signaling pathway have been linked to mesothelial cell transformation and tumor progression . P14210 and c- DB00134 are highly expressed in mesotheliomas . Phosphoinositide 3-kinase , AKT , and the downstream P42345 are involved in cell growth and survival , and they are often found to be activated in mesothelioma . p16(INK4a) and p14( Q8N726 ) are frequently inactivated in human mesothelioma , and ∼50 % of mesotheliomas contain the P35240 mutation . Molecular therapies aimed at interfering with these pathways have not improved the dismal prognosis of mesothelioma , except possibly for a small subset of patients who benefit from certain therapies . Recent studies have shown the importance of asbestos-induced inflammation in the initiation and growth of mesothelioma , and P09429 and Nalp3 inflammasome have been identified as key initiators of this process . Asbestos induces cell necrosis , causing the release of P09429 , which in turn may activate Nalp3 inflammasome , a process that is enhanced by asbestos-induced production of reactive oxygen species . P09429 and Nalp3 induce proinflammatory responses and lead to interleukin-1β and P01375 -α secretion and NF-κB activity , thereby promoting cell survival and tumor growth . Novel strategies that interfere with asbestos- and erionite-mediated inflammation might prevent or delay the onset of mesothelioma in high-risk cohorts , including genetically predisposed individuals , and/or inhibit tumor growth . The very recent discovery that germline BAP1 mutations cause a new cancer syndrome characterized by mesothelioma , uveal melanoma , and melanocytic tumors provides researchers with a novel target for prevention and early detection . Genetic variants associated with susceptibility to mother-to-child transmission of hepatitis B virus . OBJECTIVE : The mechanisms of immune tolerance to hepatitis B virus ( HBV ) in children infected perinatally or early in infancy still remain unclarified . We aimed to study the genetic variants of immune factors implicated in viral-host interaction in children who were born to HBV-positive mothers and who had different clinical outcome . METHODS : P11226 gene ( mbl2 ) codon 54 , codon 57 , and promoter 221 variants , tumor necrosis factor α ( P01375 -α ) 308G/A , and vitamin D receptor ( P11473 ) ApaI and TaqI genotypes were analyzed in three groups of children born to HBV-positive mothers : children with chronic infection ( n=33 ) , those with resolved infection ( n=36 ) , and those naive for HBV ( n=33 ) . RESULTS : P01375 -α -308G allele frequency was found to be increased in children with chronic infection compared with children who were not affected by HBV [ risk ratio ( RR ) 1.12 , 95 % confidence interval ( CI ) 1.0-1.25 ; P=0.050 ] . The P11473 ApaI A allele tended to be more frequent in children with chronic infection than in those with resolved HBV infection ( RR 1.27 , 95 % CI 0.95-1.67 ; P=0.071 ) . The P11473 ApaI α allele in ApaI and TaqI joint haplotype αT was more frequent in children with resolved infection than in those with chronic infection ( RR 1.74 , 95 % CI 0.97-3.13 ; P=0.049 ) . CONCLUSION : Our results suggest that P01375 -α and vitamin D pathways may be involved in the susceptibility to and outcome of HBV infection acquired early in life . P06850 knockout inhibits the murine innate immune responses in association with endoplasmic reticulum stress after thermal injury . BACKGROUND : Previous studies in our laboratory have demonstrated the hypothalamus destruction and adrenalectomy could blunt the innate immunity while boosting the excessive inflammation after injury . We aimed to investigate the effects of corticotrophin-releasing hormone knockout ( P06850 KO ) on the innate immune responses in macrophages as well as to elucidate the underlying mechanism . METHODS : The chemotaxis and phagocytosis activities of macrophages , bacteria translocation , plasma tumor necrosis factor ( P01375 ) -α secretion , and intestinal injury were observed in the presence of the endoplasmic reticulum stress after thermal injury in P06850 KO mice . Meanwhile , the messenger RNA ( mRNA ) and protein expression of glucose response protein 78 ( P11021 ) , X-box binding protein 1 ( P17861 ) , and activating transcription factor 6 ( P18850 ) in macrophages was also determined . RESULTS : After thermal injury , the chemotaxis and phagocytosis of peritoneal macrophages were increased , which were both reversed by P06850 gene deficiency . The gut-derived bacteria translocation to liver tissues , lung tissues and mesenteric lymph nodes was significantly strengthened in P06850 KO mice compared with P06850 wild-type littermates . Circulating P01375 -α level was increased markedly in response to thermal injury and P06850 KO further increased its secretion . Furthermore , the mRNA and protein levels of P11021 , P17861 , and P18850 in peritoneal macrophages increased , while their expressions in P06850 KO mice all decreased significantly . P06850 KO mice showed enhancement of inflammatory responses and severe tissue injuries after thermal injury . CONCLUSION : P06850 exerted immune defensive actions on immune cells and organs in the early phase of injury , suggesting that the underlying mechanisms are related to endoplasmic reticulum stress . Genetic dissociation of opiate tolerance and physical dependence in delta-opioid receptor-1 and preproenkephalin knock-out mice . Previous experiments have shown that mice lacking a functional delta-opioid receptor ( P41143 ) gene do not develop analgesic tolerance to morphine . Here we report that mice lacking a functional gene for the endogenous ligand preproenkephalin ( ppENK ) show a similar tolerance deficit . In addition , we found that the P41143 and ppENK knock-outs as well as the DB01221 receptor-deficient 129S6 inbred mouse strain , which also lacks tolerance , exhibit antagonist-induced opioid withdrawal . These data demonstrate that although signaling pathways involving ppENK , Q8IXH6 , and DB01221 receptor are necessary for the expression of morphine tolerance , other pathways independent of these factors can mediate physical dependence . Moreover , these studies illustrate that morphine tolerance can be genetically dissociated from physical dependence , and thus provide a genetic framework to assess more precisely the contribution of various cellular and molecular changes that accompany morphine administration to these processes . 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 . 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 .

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interacts_with DB09068?

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[ P35354 inhibitors -- one step forward and two steps back ] . BACKGROUND : Selective cyclooxygenase-2 ( P35354 ) inhibitors are widely used . They have no advantages in terms of efficacy , and it is not documented that they cause fewer adverse effects than conventional NSAIDs . MATERIAL AND METHODS : The adverse effects of rofecoxib , celecoxib and other NSAIDs are reviewed . Relevant literature was identified on Medline and in the reference lists in key articles . RESULTS : DB00533 and the novel P35354 inhibitors etoricoxib and valdecoxib have a higher degree of P35354 selectivity than traditional NSAIDs . Celecoxib is less P35354 selective and appears to be similar to diclofenac . DB00533 induces thromboembolic adverse effects more frequently than conventional NSAIDs . INTERPRETATION : The cardiovascular problems conferred by rofecoxib are probably a class effect and thus inducible by other selective P35354 inhibitors . Pending comprehensive safety data , caution is warranted regarding the use of these drugs . 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 . 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 . Gene expression in the anterior cingulate cortex and amygdala of adolescent marmoset monkeys following parental separations in infancy . Early life adversities are risk factors for later mood and emotional disorders . Repeated separation of infant marmosets from their parents provides a validated primate model of depression vulnerability , producing in-vivo biochemical and behavioural effects indicative of persistently altered stress reactivity and mild anhedonia . Here we report the long-term effect ( in adolescence ) of this intervention on the expression of synaptophysin , P20936 -43 , Q9P2U7 , Q9H598 , P11137 , spinophilin , and P08908 and 5- Q13049 receptors , in the anterior cingulate cortex ( ACC ; supragenual and subgenual areas ) and amygdala ( lateral , basal and central nuclei ) . These genes and regions are implicated in the response to stress or in mood disorder . The profile of P08908 receptor binding in ACC was affected by early deprivation , notably in the subgenual region , with a decrease in deep laminae but an increase in superficial laminae . Following early deprivation , spinophilin mRNA was reduced in subgenual ACC . In the amygdala , no significant effects of the manipulation were seen , but expression of several transcripts was sexually dimorphic . There were correlations between expression of some transcripts and in-vivo measurements . The results show that early deprivation in a non-human primate has a selective long-term effect on expression of genes in the ACC , particularly the subgenual area . The results differ from those reported in the hippocampus of the same animals , indicating the presence of limbic region-specific long-term molecular responses to early life stress . 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 . 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 . 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 ) Comparative molecular profiling of the PPARα/γ activator aleglitazar : Q07869 selectivity , activity and interaction with cofactors . Peroxisome proliferator-activated receptors ( PPARs ) are a family of nuclear hormone receptors that control the expression of genes involved in a variety of physiologic processes , through heterodimerization with retinoid X receptor and complex formation with various cofactors . Drugs or treatment regimens that combine the beneficial effects of PPARα and γ agonism present an attractive therapeutic strategy to reduce cardiovascular risk factors . DB08915 is a dual PPARα/γ agonist currently in phase III clinical development for the treatment of patients with type 2 diabetes mellitus who recently experienced an acute coronary event . The potency and efficacy of aleglitazar was evaluated in a head-to-head comparison with other PPARα , γ and δ ligands . A comprehensive , 12-concentration dose-response analysis using a cell-based assay showed aleglitazar to be highly potent , with EC(50) values of 5 nM and 9 nM for PPARα and PPARγ , respectively . Cofactor recruitment profiles confirmed that aleglitazar is a potent and balanced activator of PPARα and γ . The efficacy and potency of aleglitazar are discussed in relation to other dual PPARα/γ agonists , in context with the published X-ray crystal structures of both PPARα and γ . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . Ras-driven transcriptome analysis identifies aurora kinase A as a potential malignant peripheral nerve sheath tumor therapeutic target . PURPOSE : Patients with neurofibromatosis type 1 ( P21359 ) develop malignant peripheral nerve sheath tumors ( MPNST ) , which are often inoperable and do not respond well to current chemotherapies or radiation . The goal of this study was to use comprehensive gene expression analysis to identify novel therapeutic targets . EXPERIMENTAL DESIGN : Nerve Schwann cells and/or their precursors are the tumorigenic cell types in MPNST because of the loss of the P21359 gene , which encodes the P20936 protein neurofibromin . Therefore , we created a transgenic mouse model , P09543 -HRas12V , expressing constitutively active HRas in Schwann cells and defined a Ras-induced gene expression signature to drive a Bayesian factor regression model analysis of differentially expressed genes in mouse and human neurofibromas and MPNSTs . We tested functional significance of Aurora kinase overexpression in MPNST in vitro and in vivo using Aurora kinase short hairpin RNAs ( shRNA ) and compounds that inhibit Aurora kinase . RESULTS : We identified 2,000 genes with probability of linkage to nerve Ras signaling of which 339 were significantly differentially expressed in mouse and human P21359 -related tumor samples relative to normal nerves , including O14965 ( O14965 ) . O14965 was dramatically overexpressed and genomically amplified in MPNSTs but not neurofibromas . Aurora kinase shRNAs and Aurora kinase inhibitors blocked MPNST cell growth in vitro . Furthermore , an O14965 selective inhibitor , DB05220 , stabilized tumor volume and significantly increased survival of mice with MPNST xenografts . CONCLUSION : Integrative cross-species transcriptome analyses combined with preclinical testing has provided an effective method for identifying candidates for molecular-targeted therapeutics . Blocking Aurora kinases may be a viable treatment platform for MPNST . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . DB08875 as a novel therapy for renal cell carcinoma . DB08875 / DB05153 ( Exelexis , Inc. ) has demonstrated remarkable responses in kidney cancer . Preclinical results revealed P15692 , P10721 and MET inhibition in a variety of solid tumors such as thyroid , ovarian , renal , lung , liver and prostate cancers . A phase II trial demonstrated efficacy in renal cancer with a 28 % objective response rate , stable disease rate of 62 % and median progression free survival of 14.7 months . Predominant toxicities of fatigue and diarrhea were noted . Dramatic responses in bone metastases ( three of four patients ) make the agent especially valuable for palliation in a disease , where presence of bone metastases is a predictor of worse survival . DB08875 is an emerging novel agent with promising activity in advanced kidney cancer . Randomized trials are planned in comparison with standard P15692 inhibitor therapy . Defining the role of MET overexpression would help patient selection and enrich and enhance the future evaluation of this targeted novel agent . Inhibition of canine exocrine pancreatic secretion by peptide YY is mediated by P10082 -preferring P28062 receptors . It is still unclear , which receptor subtype , Q03519 and/or P28062 , mediates the inhibitory action of P10082 on exocrine pancreatic secretion . The present study was undertaken to characterize functionally the Y receptor subtype that mediates the inhibition of exocrine pancreatic secretion by peptide YY ( P10082 ) . In eight conscious dogs with chronic gastric and pancreatic fistulas , we compared the action of intravenous infusion of 200 and 400 pmol/kg/h of the Y receptor agonists P10082 1-36 , DB05004 , P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 on the pancreatic secretory response to secretin ( 20.5 pmol/kg/h ) and cerulein ( 29.6 pmol/kg/h ) . P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 were also studied by giving a fivefold dose ( 1,000 and 2,000 pmol/kg/h ) . P10082 1-36 and the P49146 agonist DB05004 significantly inhibited pancreatic secretory responses to secretin and cerulein , whereas inhibition by P01303 1-36 and the P49146 agonist P10082 13-36 was attainable only at doses of 1,000 and 2,000 pmol/kg/h . The Q03519 receptor agonist Pro34PYY 1-36 was without effect on pancreatic secretion . We conclude that in dogs the inhibition of exocrine pancreatic secretion by P10082 is mediated via P28062 receptors of a P10082 -preferring subtype . Neuropeptide Y is crucial for nutritional state-dependent regulation of maternal behavior . Lactation is indispensable for the survival of mammalian pups . However , any excess of energy expenditure for lactation over energy intake threatens the mother 's survival . Here , we report that an orexigenic molecule , neuropeptide Y ( P01303 ) , mediates nutritional state-dependent regulation of maternal behavior . After 9h of fasting , dams showed a dramatic decrease in the expression of maternal behavior . Intracerebroventricular or direct dorsal raphe nucleus ( DRN ) injection of P01303 inhibited the expression of maternal behavior in non-fasted dams . In contrast , injection of the P01303 Q03519 receptor antagonist BIBP-3226 into the DRN , in which the expression of the Q03519 receptor was confirmed in serotonergic ( 5-HT ) and GABAergic interneurons , recovered the expression of maternal behavior in fasted dams . When the pups were presented , the increase in the number of c-Fos-positive GABAergic , but not serotonergic , neurons was smaller in fasted than in non-fasted dams . These results suggest that P01303 may inhibit pup-induced activation of GABAergic neurons via the Q03519 receptor . Injection of a P08908 agonist , GABAA receptor antagonist , or GABAB receptor antagonist into the DRN induced incomplete maternal behavior in non-fasted dams . In contrast , each of a 5- Q13049 receptor agonist or a GABAB receptor agonist , but not a GABAA receptor agonist , recovered separate components of maternal behavior in fasted dams . These results suggest that P01303 inhibits both 5-HT neuronal activity and its modulation via the GABA receptor in the DRN , resulting in the suppression of maternal behavior under food-restricted conditions . 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 . DB01373 dependent activation of skeletal muscle Ca2+ release channel ( ryanodine receptor ) by calmodulin . In this study terminal cisternae vesicles from rabbit skeletal muscle were fused into planar bilayers and the effect of calmodulin on single Ca2+ release channel currents was investigated . In the presence of 10(-7) and 10(-9) M free [ Ca2+ ] , nanomolar concentrations of calmodulin activated the channel by increasing the open probability of single-channel events in a dose dependent manner . The activatory effect of calmodulin was reversed by 10 microM ruthenium red . At 10(-5) M free [ Ca2+ ] , calmodulin ( 0.1-1 microM ) inhibited channel activity . P62158 overlays were carried out using concentrations of Ca2+ similar to those used for the planar lipid bilayer assays . In the presence of 10(-7) M [ Ca2+ ] , calmodulin bound to the ryanodine receptor , to a region defined by residues 2937-3225 and 3546-3655 . These results suggest that calmodulin may activate the Ca(2+)-release channel ( ryanodine-receptor ) by interacting with binding sites localized in the central portion of the RYR protomer . Consequences of immunogenicity to the therapeutic monoclonal antibodies ReoPro and Remicade . The clinical consequences of immune antibodies generated to abciximab ( ReoPro ) and infliximab ( Remicade ) are described . DB00054 , a chimaeric Fab fragment that binds to the beta3 integrin of the P08514 /IIIa and alphavbeta3 receptors on human platelets , is approved in the US and Europe for use in percutaneous coronary intervention ( P05154 ) to prevent cardiac ischaemic complications . The effects of induced antibodies upon the safety and efficacy of repeat administration of abciximab have been evaluated in the ReoPro Re-administration Registry Study , in which 5.7 % of patients were HACA positive before re-treatment . An interim evaluation of 1000 patients has indicated that re-administration of abciximab can be accomplished in the setting of P05154 with an acceptable safety and efficacy profile . DB00065 is a chimaeric IgG1 antibody specific for human TNFalpha , and is approved in the US and Europe for the acute treatment of the signs and symptoms of Crohn 's disease and for the chronic treatment of rheumatoid arthritis ( RA ) . The incidence of antibodies to infliximab is reported to be approximately 10 % ; however , an inverse dose-immunogenicity relationship was observed , indicating that higher doses of infliximab ( > or = 3 to 10 mg/kg ) could reduce the incidence of immune antibodies . The induction of immune antibodies could also be reduced by concomitant administration of low-dose methotrexate and other immunosuppressant agents . Although antibodies to infliximab appeared to be associated with lower serum infliximab concentrations and a slightly higher incidence of infusion reactions , these immune antibodies were generally not associated with a reduction in clinical efficacy . In addition , the antibodies induced to infliximab are specific for infliximab , and do not cross-react with other currently available therapeutic antibodies . [ Clinical pharmacology of the selective P35354 inhibitors ] . The discovery of two isoforms of cyclooxygenases ( P23219 and P35354 ) has provided a new insight into the involvement of prostaglandins in the clinical effectiveness and gastrointestinal toxicity of NSAIDs . Currently , there are four selective P35354 inhibitors available in Germany : celecoxib , rofecoxib , valdecoxib and parecoxib . Orally administered rofecoxib , celecoxib and valdecoxib have been approved for the relief of symptoms of osteoarthritis and rheumatoid arthritis . DB08439 , the first selective P35354 inhibitor for parenteral administration , has been approved for the short-term treatment of post-operative pain . The clinical efficacy of the marketed P35354 inhibitors has been proved in large phase III clinical trials in comparison to both placebo and classical NSAIDs ( e.g. diclofenac , naproxen ) . The incidence of gastrointestinal complications was significantly lower than that with the non-selective NSAIDs . However , the clinical relevance of these effects was , at least in some populations of patients ( e.g. patients on low dose aspirin ) , not as high as initially expected . While not replacing less expensive classical NSAIDs , selective P35354 inhibitors provide a marked enrichment of the spectrum of anti-inflammatory and analgesic therapeutics .

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Agonists and antagonists for P2 receptors . Recent work has identified nucleotide agonists selective for P47900 , P41231 and Q15077 receptors and nucleotide antagonists selective for P47900 , Q9H244 and P51575 receptors . Selective non-nucleotide antagonists have been reported for P47900 , P41231 , Q15077 , Q9H244 , Q9BPV8 , P2X(2/3)/ P56373 and Q99572 receptors . For example , the dinucleotide P01308 37217 ( Up4dC ) potently activates the P41231 receptor , and the non-nucleotide antagonist A-317491 is selective for P2X(2/3)/ P56373 receptors . Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems , including conformationally locked moieties , have been synthesized as ligands for P2Y receptors . The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity . At P47900 ,2,4,11 receptors , there is a preference for the North conformation as indicated with ( N ) -methanocarba analogues . The P47900 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC50 of 0.95 nM . MRS2365 , an ( N ) -methanocarba analogue of 2-MeSADP , displayed potency ( EC50 ) of 0.4nM at the P47900 receptor , with > 10000-fold selectivity in comparison to Q9H244 and Q9BPV8 receptors . At Q15077 receptors there is a dramatic preference for the South conformation . Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships ( SAR ) , mutagenesis and modelling studies . Detailed three-dimensional structures of P2X receptors have not yet been proposed . 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 . [ Conditions of the primary culture for rat hepatocytes and plasminogen activator release ] . The conditions of primary culture for rat hepatocytes was investigated on the releasing effect of P00747 Activator ( PA ) . The culture method using Collagen Coated Dish ( CCD-method ) which is currently available and the ordinary culture method using Plastic Culture Dish ( P61457 -method ) were employed for that purpose in a comparative way . The effect of the addition of some supplements , that is FN , DB06692 , P01133 were also investigated . The following results were obtained . The dissociated rat hepatocytes formed a monolayer with pavementlike morphology at 24-48 hours after seeding . No difference was observed in the morphology of hepatocytes during the culture period between the two methods , although CCD-method allowed 120 hours culture , whereas P61457 -method allowed 72 hours . The PA activity was demonstrated on the hepatocytes by either culture method according to the fibrinolysis autography . The cultured hepatocytes released PA into the medium continuously as long as the viability and morphology of the cells were maintained in good state . The PA activity reached the maximum after 96 hours culture in CCD-method , whereas it reached the maximum after 48 hours in P61457 -method . The addition of DB06692 to the culture medium was not necessary for PA release in CCD-method in contrast to P61457 -method . When P01133 was discontinued in the culture medium , the release of PA was reduced in association with the occurring of morphological disintegration of hepatocytes . 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 . MafA expression and insulin promoter activity are induced by nicotinamide and related compounds in P01308 -1 pancreatic beta-cells . DB02701 has been reported to induce differentiation of precursor/stem cells toward a beta-cell phenotype , increase islet regeneration , and enhance insulin biosynthesis . Exposure of P01308 -1 beta-cells to elevated glucose leads to reduced insulin gene transcription , and this is associated with diminished binding of pancreatic duodenal homeobox factor 1 ( P52945 ) and mammalian homologue of avian MafA/l-Maf ( MafA ) . DB02701 and other low-potency poly(ADP-ribose) polymerase ( PARP ) inhibitors were thus tested for their ability to restore insulin promoter activity . The low-potency PARP inhibitors nicotinamide , 3-aminobenzamide , or PD128763 increased expression of a human insulin reporter gene suppressed by elevated glucose . In contrast , the potent P09874 inhibitors PJ34 or DB04335 -1001 had no effect on promoter activity . Antioxidants , including DB06151 , lipoic acid , or quercetin , only minimally induced the insulin promoter . Site-directed mutations of the human insulin promoter mapped the low-potency PARP inhibitor response to the C1 element , which serves as a MafA binding site . P01308 -1 cells exposed to elevated glucose had markedly reduced MafA protein and mRNA levels . Low-potency PARP inhibitors restored MafA mRNA and protein levels , but they had no affect on P52945 protein levels or binding activity . Increased MafA expression by low-potency PARP inhibitors was independent of increased MafA protein or mRNA stability . These data suggest that low-potency PARP inhibitors increase insulin biosynthesis , in part , through a mechanism involving increased MafA gene transcription . 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 . 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 . 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 . Association of the P08637 -158F/V gene polymorphism with the response to rituximab treatment in Spanish systemic autoimmune disease patients . DB00073 is being used as treatment for systemic autoimmune diseases . The objective of this study was to determine whether the genetic variant in the Fc gamma-receptor III a ( P08637 ) gene , 158F/V , contributes to the observed variation in response to rituximab in patients with systemic autoimmune diseases . DNA samples from 132 Spanish patients with different systemic autoimmune diseases receiving rituximab were genotyped for P08637 -158F/V ( rs396991 ) gene polymorphism using the TaqMan(®) allelic discrimination technology . Six months after infusion with rituximab we evaluated the response to the drug : 61 % of the patients showed a complete response , partial 27 % and 12 % did not respond to the treatment . A statistically significant difference was observed in V allele frequency between responder ( 38 % ) and nonresponder ( 16 % ) patients ( p=0.01 ; odds ratio [OR]=3.24 , 95 % confidence interval [ CI ] 1.17-11.1 ) . DB00073 was also more effective in V allele carriers ( 94 % ) than in homozygous FF patients ( 81 % ) : p=0.02 ; OR=3.96 , 95 % CI 1.10-17.68 . These results suggest that P08637 -158F/V ( rs396991 ) gene polymorphism play a role in the response to rituximab in autoimmune diseases . Validation of these findings in independent cohorts is warranted . Sustained increase of PKA activity in the postcommissural putamen of dyskinetic monkeys . Levodopa-induced dyskinesias ( LID ) are a frequent complication of Parkinson 's disease pharmacotherapy that causes significant disability and narrows the therapeutic window . Pharmacological management of LID is challenging partly because the precise molecular mechanisms are not completely understood . Here , our aim was to determine molecular changes that could unveil targetable mechanisms underlying this drug complication . We examined the expression and downstream activity of dopamine receptors ( DR ) in the striatum of 1-methyl-4-phenyl-1,2,3,6 tetrahydropiridine ( MPTP ) -lesioned monkeys with and without DB01235 treatment . Four monkeys were made dyskinetic and other four received a shorter course of DB01235 and did not develop LID . Our results show that DB01235 treatment induces an increase in P14416 and P35462 expression in the postcommissural putamen , but only P35462 is correlated with the severity of LID . Dyskinetic monkeys show a hyperactivation of the canonical P21728 -signaling pathway , measured by an increased phosphorylation of protein kinase A ( PKA ) and its substrates , particularly Q9UD71 . In contrast , activation of the P14416 -signaling pathway , visible in the levels of Akt phosphorylated on Thr308 and GSK3β on Ser9 , is associated with DB01235 treatment , independently of the presence of dyskinesias . Our data clearly demonstrate that dyskinetic monkeys present a dysregulation of the P35462 receptor and the P21728 pathway with a sustained increase of PKA activity in the postcommissural putamen . Importantly , we found that all signaling changes related to long-term DB01235 administration are exquisitely restricted to the postcommissural putamen , which may be related to the recurrent failure of pharmacological approaches . P00747 -induced aggregation of PANC-1 cells requires conversion to plasmin and is inhibited by endogenous plasminogen activator inhibitor-1 . PANC-1 cells express proteinase-activated receptors ( PARs ) -1 , -2 , and respond to their activation by transient elevation of cytosolic [ Ca(2+) ] and accelerated aggregation ( Wei et al. , 2006 , J Cell Physiol 206:322-328 ) . We studied the effect of plasminogen ( Q9UQ90 ) , an inactive precursor of the P25116 -activating protease , plasmin ( PN ) on aggregation of pancreatic adenocarcinoma ( PDAC ) cells . A single dose of Q9UQ90 time- and dose-dependently promoted PANC-1 cells aggregation in serum-free medium , while PN did not . PANC-1 cells express urokinase plasminogen activator ( uPA ) , which continuously converted Q9UQ90 to PN . This activity and Q9UQ90 -induced aggregation were inhibited by the uPA inhibitor amiloride . Q9UQ90 -induced aggregation was also inhibited by alpha-antiplasmin and by the PN inhibitor epsilon-aminocaproic acid ( DB00513 ) . Direct assay of uPA activity revealed very low rate , markedly enhanced in the presence of Q9UQ90 . Moreover , in Q9UQ90 activator inhibitor 1-deficient PANC-1 cells , uPA activity and Q9UQ90 -induced aggregation were markedly potentiated . Two additional human PDAC cell lines , MiaPaCa and Colo347 , were assayed for Q9UQ90 -induced aggregation . Both cell lines responded by aggregation and exhibited Q9UQ90 -enhanced uPA activity . We hypothesized that the continuous conversion of Q9UQ90 to PN by endogenous uPA is limited by PN 's degradation and negatively controlled by endogenously produced P05121 . Indeed , we found that PANC-1 cells inactivate PN with t1/2 of approximately 7 h , while the continuous addition of PN promoted aggregation . Our data suggest that PANC-1 cells possess intrinsic , P05121 -sensitive mechanism for promotion of aggregation and differentiation by prolonged exposure to Q9UQ90 and , possibly , additional precursors of PARs agonists . Alteration of synaptic activity-regulating genes underlying functional improvement by long-term exposure to an enriched environment in the adult brain . BACKGROUND : Housing animals in an enriched environment ( EE ) enhances behavioral function . However , the mechanism underlying this EE-mediated functional improvement and the resultant changes in gene expression have yet to be elucidated . OBJECTIVES : We attempted to investigate the underlying mechanisms associated with long-term exposure to an EE by evaluating gene expression patterns . METHODS : We housed 6-week-old CD-1 ( ICR ) mice in standard cages or an EE comprising a running wheel , novel objects , and social interaction for 2 months . Motor and cognitive performances were evaluated using the rotarod test and passive avoidance test , and gene expression profile was investigated in the cerebral hemispheres using microarray and gene set enrichment analysis ( GSEA ) . RESULTS : In behavioral assessment , an EE significantly enhanced rotarod performance and short-term working memory . Microarray analysis revealed that genes associated with neuronal activity were significantly altered by an EE . GSEA showed that genes involved in synaptic transmission and postsynaptic signal transduction were globally upregulated , whereas those associated with reuptake by presynaptic neurotransmitter transporters were downregulated . In particular , both microarray and GSEA demonstrated that EE exposure increased opioid signaling , acetylcholine release cycle , and postsynaptic neurotransmitter receptors but decreased Na+ / Cl- -dependent neurotransmitter transporters , including dopamine transporter Slc6a3 in the brain . Western blotting confirmed that Q01959 , Q9UD71 ( Q9UD71 ) , and Q9H244 were largely altered in a region-specific manner . CONCLUSION : An EE enhanced motor and cognitive function through the alteration of synaptic activity-regulating genes , improving the efficient use of neurotransmitters and synaptic plasticity by the upregulation of genes associated with postsynaptic receptor activity and downregulation of presynaptic reuptake by neurotransmitter transporters . Thrombin ( P25116 ) -induced proliferation in astrocytes via MAPK involves multiple signaling pathways . Protease-activated receptors ( PARs ) , newly identified members of G protein-coupled receptors , are widely distributed in the brain . Thrombin evokes multiple cellular responses in a large variety of cells by activating P25116 , -3 , and -4 . In cultured rat astrocytes we investigated the signaling pathway of thrombin- and PAR-activating peptide ( PAR-AP ) -induced cell proliferation . Our results show that PAR activation stimulates proliferation of astrocytes through the P29323 pathway . Thrombin stimulates P27361 /2 phosphorylation in a time- and concentration-dependent manner . This effect can be fully mimicked by a specific P25116 -AP but only to a small degree by PAR-3-AP and Q96RI0 -AP . P55085 -AP can induce a moderate P27361 /2 activation as well . Thrombin-stimulated P27361 /2 activation is mainly mediated by P25116 via two branches : 1 ) the PTX-sensitive G protein/(betagamma-subunits)-phosphatidylinositol 3-kinase branch , and 2 ) the G(q)- P98160 - ( InsP(3) receptor ) /Ca2+ -PKC pathway . Thrombin- or P25116 -AP-induced P29323 activation is partially blocked by a selective P01133 receptor inhibitor , AG1478 . Nevertheless , transphosphorylation of P01133 receptor is unlikely for P27361 /2 activation and is certainly not involved in P25116 -induced proliferation . The metalloproteinase mechanism involving transactivation of the P01133 receptor by released heparin-binding P01133 was excluded . P01133 receptor activation was detected by the receptor autophosphorylation site , tyrosine 1068 . Our data suggest that thrombin-induced mitogenic action in astrocytes occurs independently of P01133 receptor transphosphorylation . DB00758 resistance " Live " - the risk of stent thrombosis should be evaluated before procedures . Every year , millions of people undergo percutaneous coronary intervention ( P05154 ) with intracoronary stent implantation . A patient from the PRAGUE-8 trial ( Optimal pre- P05154 clopidogrel loading : 600 mg before every coronary angiography vs. 600 mg in the cath-lab only for P05154 patients ) is described who suffered from acute stent thrombosis . This patient did not have any relevant inhibition of platelet activation even after the 600 mg dose of clopidogrel . Dose uptitration would have been ineffective . New Q9H244 receptor inhibitors are desperately needed . In the light of recently published data , the use of prasugrel may be considered as an alternative . DB09078 : first global approval . DB09078 ( Lenvima™ ) is a multitargeted receptor kinase inhibitor that inhibits the kinase activities of vascular endothelial-derived growth factor receptors 1 , 2 and 3 , fibroblast growth factor receptors 1 , 2 , 3 and 4 , platelet-derived growth factor receptor α , P07949 and P10721 . In addition to their role in normal cellular function , these kinases have been implicated in pathogenic angiogenesis , tumour growth and cancer progression . DB09078 is being developed by Eisai Co . Ltd for the treatment of solid tumours , primarily for differentiated thyroid cancer , and other malignancies . A capsule formulation of the drug has received approval in the USA for use in locally recurrent or metastatic , progressive , radioactive iodine-refractory differentiated thyroid cancer . DB09078 is in pre-registration for this indication in the EU , Australia , Brazil , Canada , Japan , South Korea , Russia , Singapore and Switzerland , and is in phase 3 development in Argentina , Chile and Thailand . DB09078 has orphan designation in the EU and Japan for use in differentiated thyroid cancer . In addition , an ongoing global , phase 3 trial is evaluating the use of lenvatinib as first-line treatment in unresectable hepatocellular carcinoma . This article summarizes the milestones in the development of lenvatinib leading to this first approval in locally recurrent or metastatic , progressive , radioactive iodine-refractory differentiated thyroid cancer . Alterations of respiratory chain complexes in sporadic pheochromocytoma . DB00139 dehydrogenase ( SDH ) has been associated with carcinogenesis in hereditary pheochromocytoma ( PC ) and paraganglioma . We investigated if a similar association applies to sporadic pheochromocytoma . No genetic alteration was found in the P21912 , Q99643 or O14521 genes of sporadic PC . However , in eight of nine sporadic PCs the SDH activity was , on average , reduced by 40 % ; moreover , the activities of the other oxidative phosphorylation ( OXPHOS ) complexes and citrate synthase were significantly lower compared to normal kidney tissue . Furthermore , immunohistochemical staining revealed a significant down-regulation of respiratory chain complexes . Since no pathogenic mutations were detected in the von Hippel-Lindau ( P40337 ) gene , we can rule out that P40337 deficiency is causing the general reduction of OXPHOS enzymes observed in the PCs investigated . In contrast to the single enzyme defects found in a subset hereditary PCs , a more generalized reduction of mitochondrial respiration seems to be present in most sporadic PCs . Strikingly , one of the nine PCs showed specific loss of complex I and a compensatory up-regulation of complexes II-V , which is a phenotype usually characteristic of oncocytic tumors . [ Expression of chemokine receptors by cancer cells ] . Chemokine receptors are involved in the trafficking of leucocytes . It has been shown recently that chemokine receptors ( P61073 , P32248 ... ) are expressed by breast cancer cells and could be involved in the metastasis phenomenon . Since this discovery , new data have been generated in this topic . These data confirm that chemokine receptors are involved in the occurrence of metastases , suggest that their expression is regulated by genes involved in the carcinogenesis and/or angiogenesis ( P40337 ... ) and that inhibitors could be of interest in the treatment of breast cancer . In this paper , we will review all these data and will discuss the perspectives that opens this new concept . Functional expression of P61073 in somatotrophs : P48061 activates GH gene , GH production and secretion , and cellular proliferation . The interaction of chemokine ( C-X-C motif ) ligand 12 ( P48061 ) and its receptor P61073 may play an important role in the regulation of anterior pituitary function . In this study , we investigated the expression of P48061 and P61073 and their role in normal rat pituitary and GH-producing GH3 tumor cell line . RT-PCR analysis and immunohistochemistry revealed that P61073 was expressed in normal rat anterior pituitary and GH3 tumor cells . Double immunofluorescent staining showed the complete colocalization of P61073 with GH in rat pituitary , indicating that P61073 is specifically expressed in rat somatotrophs . Using rat primary pituitary cell cultures and GH releasing hormone receptor expressing stable GH3 cells ( GH3- Q02643 ) , we evaluated the function of P48061 compared with P01286 . P48061 stimulated GH gene activation in both primary rat anterior pituitary cells and GH3- Q02643 cells . P48061 also stimulated GH secretion from primary rat pituitary cells in a dose-dependant manner . BrdU incorporation was increased in response to P48061 addition in GH3 cell culture , indicating P48061 -induced cell proliferation . P48061 -dependent phosphorylation of P27361 /2 was also confirmed by western blot analysis , supporting the evidence that MAPK is an intracellular mediator of P48061 / P61073 interaction in GH3 cell proliferation . In conclusion , these results indicate that P48061 / P61073 interaction plays an important role in GH production , secretion , and the proliferation of somatotrophs . 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 . 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 . Genetic risk factors for infection in patients with early rheumatoid arthritis . We analyzed clinical and genetic factors contributing to infections in 457 subjects with early rheumatoid arthritis ( RA ) enrolled in a prospective , 1-year clinical trial of methotrexate and the P01375 inhibitor etanercept . Subjects were genotyped for the following single nucleotide polymorphisms ( SNPs ) : ( P01375 -308 , -238 , and + 488 ) ; lymphotoxin-alpha ( P01374 ) ( P01374 + 249 , + 365 , and + 720 ) ; and Fc gamma receptors P12318 131 H/R ; P08637 176 F/V ; and O75015 NA 1/2 and genotypes were correlated with infections . At least one O94763 was noted in 52 % of subjects ( 99/191 ) with the NA2/NA2 genotype of the neutrophil-specific O75015 gene , compared to 42 % ( 77/181 ) of those with the P10909 genotype and 39 % ( 23/59 ) of those with the NA1/NA1 genotype ( P = 0.038 ) . Urinary tract infection ( UTI ) was associated with the P01375 -238 A ( odds ratio(OR) 2.56 , 95 % confidence interval ( CI ) 1.05-6.25 ) and P01374 +365 C ( OR 1.73 , 95 % CI 1.07-2.79 ) alleles , and marginally with the P08637 F allele ( OR 1.72 , 95 % CI 0.99-3.00 ) . There was a striking linear correlation between UTI and the number of risk alleles defined by these three SNPs ( P < 0.001 ) , suggesting an additive effect on susceptibility . These findings have important implications for the role of genetics in susceptibility to bacterial and viral infections . 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 . [ 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 . Comparison of three GPCR structural templates for modeling of the Q9H244 nucleotide receptor . The P2Y(12) receptor ( P2Y(12)R ) is an ADP-activated G protein-coupled receptor ( GPCR ) that is an important target for antithrombotic drugs . Three homology models of P2Y(12)R were compared , based on different GPCR structural templates : bovine rhodopsin ( bRHO ) , human A(2A) adenosine receptor ( A(2A)AR ) , and human P61073 ( P61073 ) . By criteria of sequence analysis ( 25.6 % identity in transmembrane region ) , deviation from helicity in the second transmembrane helix ( TM2 ) , docked poses of ligands highlighting the role of key residues , accessibility of a conserved disulfide bridge that is reactive toward irreversibly-binding antagonists , and the presence of a shared disulfide bridge between the third extracellular loop ( EL3 ) and the N-terminus , the P61073 -based model appeared to be the most consistent with known characteristics of P2Y(12)R . The docked poses of agonist 2MeSADP and charged anthraquinone antagonist PSB-0739 in the binding pocket of P2Y(12)R-CXC agree with previously published site-directed mutagenesis studies of Arg256 and Lys280 . A sulfonate at position 2 of the anthraquinone core created a strong interaction with the Lys174(EL2) side chain . The docking poses of the irreversibly-binding , active metabolite ( existing as two diastereoisomers in vivo ) of the clinically utilized antagonist DB00758 were compared . The free thiol group of the 4S diastereoisomer , but not the 4R isomer , was found in close proximity ( ~4.7 Å ) to the sulfur atom of a disulfide bridge involving Cys175 , suggesting greater activity in covalent binding . Therefore , ligand docking to the P61073 -based model of the P2Y(12)R predicted poses of both reversibly and irreversibly-binding small molecules , consistent with observed pharmacology and mutagenesis studies .

MH_train_1536

MH_train_1536

MH_train_1536

interacts_with DB00559?

multiple_choice

Dual endothelin receptor antagonism prevents remodeling of resistance arteries in diabetes . Vascular remodeling , characterized by extracellular matrix deposition and increased media-to-lumen ( M/L ) ratio , contributes to the development of microvascular complications in diabetes . We have previously shown in type 2 diabetic Goto-Kakizaki ( GK ) rats that selective P25101 receptor blockade prevents medial thickening of mesenteric arteries via regulation of matrix metalloproteases ( MMP ) , whereas selective ETB receptor blockade augments this thickening . The goal of this study was to determine the effect of combined P25101 and ETB receptor blockade on resistance vessel remodeling . Vessel structure , MMP activity , and extracellular matrix proteins were assessed in control Wistar and diabetic GK rats treated with vehicle or DB00559 ( 100 mg/kg per day ) for 4 weeks ( n = 7-9 per group ) . DB00559 completely prevented the increase in M/L ratio and P08253 activity in diabetes but paradoxically increased M/L ratio and MMP activation in control animals . Collagenase ( P45452 ) activity and protein levels were significantly decreased in diabetes . Accordingly , collagen deposition was augmented in GK rats . Dual ET receptor antagonism improved enzyme activity and normalized P45452 levels in diabetic animals but blunted P45452 activity in control animals . In summary , current findings suggest that diabetes-mediated remodeling of resistance arteries is prevented by dual blockade of P25101 and ETB receptors and that the relative role of ET receptors in the regulation of vascular structure differs in the control and disease states . Effect of endothelin receptor antagonists on non-muscle matrix compaction in a cell culture vasospasm model . P05305 ( ET-1 ) , a potent vascular smooth muscle constrictor , is one of the possible spasmogens in cerebral vasospasm . However , the role of ET-1 in non-muscle compaction ( another aspect of the pathogenesis of cerebral vasospasm ) has not been reported . This study was undertaken to demonstrate the effect of ET-1 , as well as erythrocyte lysate and bloody cerebrospinal fluid ( P04141 ) , on fibroblast populated collagen lattice ( FPCL ) compaction . Human dermal fibroblasts were used to form FPCL . The concentration-dependent effect of ET-1 was examined in the absence and presence of an P25101 receptor antagonist ( BQ-485 ) , or an ETB receptor antagonist ( BQ-788 ) , or both . FPCL compaction was determined by measuring reduction of areas over five days following treatment . To compare the effect of ET-1 on lattice compaction , erythrocyte lysate and bloody P04141 obtained from a cerebral vasospasm patient were also tested . We found that ET-1 increased FPCL compaction in a concentration-dependent ( but not time-dependent ) manner . Erythrocyte lysate produced the strongest compaction , however , without time-dependence . Bloody P04141 promoted FPCL compaction in a time-dependent fashion . Compaction induced by ET-1 was inhibited by BQ-485 but not by BQ-788 . We concluded that ET-1 promotes FPCL compaction by activation of P25101 receptors . Other components in bloody P04141 or erythrocytes may also contribute to FPCL compaction . Progress in studies of huperzine A , a natural cholinesterase inhibitor from Chinese herbal medicine . DB04864 ( HupA ) , a novel alkaloid isolated from the Chinese herb Huperzia serrata , is a potent , highly specific and reversible inhibitor of acetylcholinesterase( P22303 ) . Compared with tacrine , donepezil , and rivastigmine , HupA has better penetration through the blood-brain barrier , higher oral bioavailability , and longer duration of P22303 inhibitory action . HupA has been found to improve cognitive deficits in a broad range of animal models . HupA possesses the ability to protect cells against hydrogen peroxide , beta-amyloid protein ( or peptide ) , glutamate , ischemia and staurosporine-induced cytotoxicity and apoptosis . These protective effects are related to its ability to attenuate oxidative stress , regulate the expression of apoptotic proteins Bcl-2 , Bax , P04637 , and caspase-3 , protect mitochondria , upregulate nerve growth factor and its receptors , and interfere with amyloid precursor protein metabolism . Antagonizing effects of HupA on N-methyl-D-aspartate receptors and potassium currents may also contribute to its neuroprotection as well . Pharmacokinetic studies in rodents , canines , and healthy human volunteers indicated that HupA was absorbed rapidly , distributed widely in the body , and eliminated at a moderate rate with the property of slow and prolonged release after oral administration . Animal and clinical safety tests showed that HupA had no unexpected toxicity , particularly the dose-limiting hepatotoxicity induced by tacrine . The phase IV clinical trials in China have demonstrated that HupA significantly improved memory deficits in elderly people with benign senescent forgetfulness , and patients with Alzheimer disease and vascular dementia , with minimal peripheral cholinergic side effects and no unexpected toxicity . HupA can also be used as a protective agent against organophosphate intoxication . Clinical correlates of promoter hypermethylation of four target genes in head and neck cancer : a cooperative group correlative study . PURPOSE : Promoter hypermethylation is a well-documented mechanism for tumor-specific alteration of suppressor gene activity in human malignancy including head and neck cancer ( HNC ) . The abrogation of specific suppressor gene activity may influence tumor behavior and clinical outcome . In this study we examined methylation of P43146 , Q12756 , P24530 , and p16(INK4a) in a large cohort of HNC patients from Eastern Cooperative Group ( ECOG ) 4393/Radiation Therapy Oncology Group ( RTOG ) 9614 to identify clinical correlates of methylation of these genes . EXPERIMENTAL DESIGN : Methylation was assessed by quantitative methylation-specific PCR in DNA from tumor specimens and was considered as a continuous and a binary variable . Clinical data including demographics , stage , risk factor exposure , treatment , and outcome were collected by ECOG and RTOG . Methylation status was also correlated with mutation of P04637 ( previously reported ) and human papilloma virus status . RESULTS : Methylation results were available for 368 cases , 353 of which also have p53 mutation status . At least one methylation event was present in all tumors . In multivariate analysis of the entire cohort , methylation of p16 was associated with decreased survival ( HR = 1.008 ; P = 0.045 ) . However , in tumors with disruptive P04637 mutation ( poor prognostic group ) , the additional presence of methylation of p16 was protective ( P = 0.019 considering p16 methylation as a continuous variable ) . CONCLUSION : Methylation of tumor-related genes contributes to the biological behavior of HNC and influences overall survival in conjunction with other known prognostic molecular events . The effect of endoscopic retrograde cholangiopancreatography on the serum Q14116 and erythrocytes antioxidative capacity in biliary obstructive jaundice . INTRODUCTION : Obstructive jaundice is an important clinical problem . It may cause transient hemolysis and shortened erythrocyte life span as well as cytokine induction . An increase in lipid peroxidation has been noted as evidence of oxidative damage in red cells due to cholestasis . The influence of endoscopic retrograde cholangiopancreatography ( ERCP ) , mechanical lithotrepsy and stone extraction on the antioxidative capacity of the erythrocyte and immune response is still unclear . METHODS : Superoxide dismutase ( SOD ) , catalase ( CAT ) and glutathione ( DB00143 ) content of red blood cells ( RBC ) , and serum interleukin ( Q14116 ) were measured in 20 patients with calcular obstructive jaundice before and 4 weeks after ERCP intervention and compared with 10 matched healthy volunteers . RESULTS : A significant decrease ( p < 0.05 ) in SOD and CAT activities and glutathione concentration but a significant increase ( p < 0.05 ) in serum Q14116 were observed in cholestatic patients compared with the healthy control and were significantly correlated with variable of hepatic dysfunction ( alanine aminotransferase ( ALT ) , aspartate aminotransferase ( Q9NRA2 ) , total bilirubin , alkaline phosphatase ( ALP ) and gamma glutamyl transferase ( P19440 ) . After ERCP , serum Q14116 and antioxidant capacity of red blood cells were significantly improved and returned to normal concentration ( p < 0.05 ) . CONCLUSIONS : In biliary obstruction , serum Q14116 is increased and antioxidative capacity is decreased , and have a direct correlation with biochemical markers of liver injury . After ERCP intervention , the altered antioxidative capacity as well as serum Q14116 was completely restored to normal . HIF prolyl hydroxylase-2 inhibition diminishes tumor growth through matrix metalloproteinase-induced TGFβ activation . A right amount of oxygen and nutrients is essential for a tumor to develop . The role of oxygen dependent pathways and their regulators is therefore of utmost importance although little is known about the detailed impact they can have . Recently we have shown that inhibition of the oxygen sensor Q9GZT9 in tumor cells blocks tumor growth due to the anti-proliferative activity of TGFβ . In this study , we refined these results by comparing different shPHD2 sequences in depth in the early phase of tumor growth . Our findings also reveal an intriguing role for P08253 and P50281 in these settings , as these activated proteases display an anti-proliferative characteristic through the activation of downstream TGFβ targets . In conclusion , Q9GZT9 inhibition is essential for the regulation of the anti-tumoral activity in mouse tumor cells and might bring some new insight in our understanding of tumor growth inhibition . Differential expression of the rat gamma-glutamyl transpeptidase gene promoters along with differentiation of hepatoblasts into biliary or hepatocytic lineage . gamma-Glutamyl transpeptidase ( P19440 ) , a major enzyme of glutathione ( DB00143 ) homeostasis , is often used as a biliary marker to follow the differentiation of hepatic precursor cells . The expression of the P19440 gene is driven by different promoters and yields multiple mRNAs , depending on the cell type or the stage of differentiation . In the present study , we analyzed the P19440 mRNA expression pattern by quantitative reverse transcriptase-polymerase chain reaction or by in situ hybridization i ) in the liver , in vivo , at early stages of development ; ii ) in oval cells , which proliferate and differentiate into hepatocytes in response to galactosamine injury in vivo ; and finally , iii ) during hepatoblast differentiation , in vitro . We show that P19440 gene transcription originates from promoters P09131 , P4 , and Q15084 in rat hepatic precursor cells . Differentiation of these cells induces profound alterations in P19440 gene expression , leading to extinction of promoters P4 and Q15084 , when they differentiate into the hepatocytic pathway , and to extinction of promoters P09131 and Q15084 when they differentiate into the biliary pathway . This diversity in P19440 mRNA expression provides unique molecular probes to follow hepatic precursor cell differentiation . Furthermore , the identification of factors governing P19440 Q15084 and P4 promoter expression should provide further insight into the molecular events that occur as the liver precursor cell differentiates into the hepatic lineages . 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 . 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 . Disruption of hSWI/SNF complexes in T cells by P42768 mutations distinguishes X-linked thrombocytopenia from Wiskott-Aldrich syndrome . Wiskott-Aldrich syndrome ( P42768 ) , an immunodeficiency disorder , and X-linked thrombocytopenia ( XLT ) , a bleeding disorder , both arise from nonsynonymous mutations in P42768 , which encodes a hematopoietic-specific P42768 . Intriguingly , XLT evolves into P42768 in some patients but not in others ; yet the biological basis for this cross-phenotype ( CP ) effect remains unclear . Using human T-helper ( TH ) cells expressing different disease-causing P42768 mutations , we demonstrated that hSWI/SNF-like complexes require nuclear- P42768 to execute their chromatin-remodeling activity at promoters of P42768 -target , immune function genes during Q8IXH7 differentiation . Hot-spot P42768 mutations Thr45Met and Arg86Cys , which result in XLT-to- P42768 disease progression , impair recruitment of P51531 - but not P51532 -enriched O75531 complexes to P01579 and Q9UL17 promoters . Moreover , promoter enrichment of histone H2A.Z and its catalyzing enzyme Q96L91 are both impaired . Consequently , activation of Notch signaling , a P51531 -regulated event , and its downstream effector NF-κB are both compromised , along with decreased accessibility of nucleosomal DNA and inefficient transcription-elongation of P42768 -target Q8IXH7 genes . In contrast , patient mutations Ala236Gly and Arg477Lys that manifest in XLT without progressing to P42768 do not disrupt chromatin remodeling or transcriptional reprogramming of Q8IXH7 genes . Our study defines an indispensable relationship between nuclear- P42768 - and hSWI/SNF-complexes in gene activation and reveals molecular distinctions in TH cells that might contribute to disease severity in the XLT/ P42768 clinical spectrum . 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 . DB08890 - a secretagogue and antihyperalgesic agent - what next ? Ongoing clinical trials suggest that linaclotide , a first-in-class , 14-amino acid peptide guanylate cyclase-C ( P25092 ) receptor agonist and intestinal secretagogue is an effective treatment for chronic constipation . A study in this issue of the Journal suggests that linaclotide also has antihyperalgesic effects in three common rat models of inflammation- and stress-induced hypersensitivity ( i.e. , acute trinitrobenzene sulfonic acid colitis , water avoidance stress [ P42768 ] , and restraint-induced stress ) but not in naïve animals . In mice , linaclotide at least partly reduces hyperalgesia via P25092 receptors . Dose-effect relationships of linaclotide were complicated and non-linear . This viewpoint discusses human clinical trials with linaclotide and the results of this study . Potential mechanisms and clinical significance of these findings are explored . Collectively , these data suggest that P25092 receptors exert other , as yet poorly understood , effects on gastrointestinal sensitivity in conditions associated with inflammation and/or stress-induced increased intestinal permeability . However , the data need to be confirmed in humans and in long-term animal models . Further studies are also necessary to elucidate the mechanisms as these effects can not be explained by linaclotide 's known effects on epithelial P25092 receptors . 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 . Desensitization and internalization of endothelin receptor A : impact of G protein-coupled receptor kinase 2 ( P25098 ) -mediated phosphorylation . Endothelin receptor A ( P25101 ) , a G protein-coupled receptor , mediates endothelin signaling , which is regulated by P25098 . Three DB00133 and seven DB00156 residues recently proven to be phosphoacceptor sites are located in the C-terminal extremity ( CTE ) of the receptor following its palmitoylation site . We created various phosphorylation-deficient P25101 mutants . The phospholipase C activity of mutant receptors in P29320 -293 cells was analyzed during continuous endothelin stimulation to investigate the impact of phosphorylation sites on P25101 desensitization . Total deletion of phosphoacceptor sites in the CTE affected proper receptor regulation . However , proximal and distal phosphoacceptor sites both turned out to be sufficient to induce WT-like desensitization . Overexpression of the Gαq coupling-deficient mutant P25098 -D110A suppressed P25101 -WT signaling but failed to decrease phospholipase C activity mediated by the phosphorylation-deficient mutant P25101 -6PD . In contrast , P25098 -WT acted on both receptors , whereas the kinase-inactive mutant P25098 -D110A/K220R failed to inhibit signaling of P25101 -WT and P25101 -6PD . This demonstrates that P25101 desensitization involves at least two autonomous P25098 -mediated components : 1 ) a phosphorylation-independent signal decrease mediated by blocking of Gαq and 2 ) a mechanism involving phosphorylation of DB00133 and DB00156 residues in the CTE of the receptor in a redundant fashion , able to incorporate either proximal or distal phosphoacceptor sites . High level transfection of P25098 variants influenced signaling of P25101 -WT and P25101 -6PD and hints at an additional phosphorylation-independent regulatory mechanism . Furthermore , internalization of mRuby-tagged receptors was observed with P25101 -WT and the phosphorylation-deficient mutant P25101 -14PD ( lacking 14 phosphoacceptor sites ) and turned out to be based on a phosphorylation-independent mechanism . 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 . Thrombin antithrombin complex and Q14116 serum levels in stroke patients . The complex picture of inflammation and coagulation alterations comes to life in acute stroke phases . Increasing evidence points to a strong interaction and extensive crosstalk between the inflammation and coagulation systems : the interest towards this relationship has increased since recent experimental research showed that the early administration of antithrombin III ( P01008 ) decreases the volume of ischemia in mice and might be neuroprotective , playing an antiinflammatory role.We aimed to establish the extent of the relationship among markers of inflammation ( P04271 and Q14116 ) and procoagulant and fibrinolytic markers ( P01008 , thrombin-antithrombin III complex ( TAT ) , Fibrin Degradation Products ( Q9NRC9 ) , D-dimer ) in 13 comatose patients affected by focal cerebral ischemia.Plasma levels of TAT , D-dimer and Q9NRC9 , Q14116 and P04271 were increased . Q14116 and P04271 high serum levels in ischemic patients suggest an early activation of the inflammatory cascade in acute ischemic injury.The basic principles of the interaction between inflammatory and coagulation systems are revised , from the perspective that simultaneous modulation of both coagulation and inflammation , rather than specific therapies aimed at one of these systems could be more successful in stroke therapy . 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 . Could treatment with arundic acid ( DB05343 ) increase vulnerability for depression ? Arundic acid ( DB05343 ) is believed to be neuroprotective because of its actions on glia cells ; i.e. , its inhibitory effects on the synthesis of a calcium-binding protein P04271 . DB05343 is undergoing clinical trials for the treatment of patients with stroke and Alzheimer 's disease . Recent clinical studies point to a pervasive comorbidity of depression with stroke and Alzheimer 's disease . Previously , P04271 has been implicated in the pathobiological mechanisms of depression . Preclinical studies have shown that antidepressant treatment significantly increases brain P04271 . Here we hypothesize that available data that link P04271 with depression , along with the proposed inhibitory action of DB05343 on P04271 synthesis , indicate that this compound could increase vulnerability for depression in patients at risk for this disorder , and we propose that evaluation of patients with stroke and Alzheimer 's disease for the presence of depression should be routine in clinical trials employing DB05343 . Although it may be open for discussion whether the neuroprotective effects of DB05343 are exclusively due to its inhibition of P04271 synthesis , the latter action of DB05343 warrants studies of the effects of this drug in the pathobiology of depression . Histochemical studies on endothelin and the endothelin-A receptor in the hypothalamus . By use of a rabbit polyclonal antibody specific to the endothelin-A ( P25101 ) receptor , we analyzed the distribution of neurons containing P25101 receptors in rat brain . Almost all A1-A7 noradrenergic neurons , Q92854 -A16 and retinal amacrine ( A17 ) dopaminergic neurons ( except the P41732 group ) , and P01024 adrenergic neurons contained P25101 receptors . In addition , hypothalamic magnocellular ( noncatecholaminergic ) neurons also showed immunoreactivity to P25101 receptors , whereas the staining intensity was low . These observations were confirmed by analyzing the expression of c-fos in the hypothalamus after central injection of ET isopeptides . We also examined immunohistochemically the distribution of big ETs and mature ETs in the hypothalamus . The paraventricular and supraoptic magnocellular neurons in the hypothalamus were immunopositive for big ET-1 and mature ET-1 . However , their nerve fibers were immunostained only with antibody to big ET-1 . The observations on big ET-1 were altered by colchicine treatment . Considering all the evidence , mature ET-1 may regulate neurotransmission in the hypothalamic region , whereas big ET-1 is secreted from nerve terminals into the vessels as a neurohormone . It is also concluded that ET , one of the " brain-vascular peptides , " has a close relationship with the catecholamine neuron system in the brain . 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 . DB00126 is dispensable for oxygen sensing in vivo . Prolyl-4-hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl-4-hydroxylase domain ( P20941 ) enzymes requires oxygen and 2-oxoglutarate as cosubstrates with iron(II) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo(-/-) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo(-/-) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum- and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF-1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl(2) . A Cys201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione .

MH_train_1537

MH_train_1537

MH_train_1537

interacts_with DB00502?

multiple_choice

Blocking dopamine D2 receptors by haloperidol curtails the beneficial impact of calorie restriction on the metabolic phenotype of high-fat diet induced obese mice . Calorie restriction is the most effective way of expanding life-span and decreasing morbidity . It improves insulin sensitivity and delays the age-related loss of dopamine receptor D(2) ( P14416 ) expression in the brain . Conversely , high-fat feeding is associated with obesity , insulin resistance and a reduced number of P14416 binding sites . We hypothesised that the metabolic benefit of calorie restriction involves the preservation of appropriate P14416 transmission . The food intake of wild-type C57Bl6 male mice was restricted to 60 % of ad lib. intake while they were treated with the P14416 antagonist haloperidol or vehicle using s.c. implanted pellets . Mice with ad lib. access to food receiving vehicle treatment served as controls . All mice received high-fat food throughout the experiment . After 10 weeks , an i.p. glucose tolerance test was performed and , after 12 weeks , a hyperinsulinaemic euglycaemic clamp . Hypothalamic P14416 binding was also determined after 12 weeks of treatment . Calorie-restricted ( CR ) vehicle mice were glucose tolerant and insulin sensitive compared to ad lib . ( AL ) fed vehicle mice . CR mice treated with haloperidol were slightly heavier than vehicle treated CR mice . DB00502 completely abolished the beneficial impact of calorie restriction on glucose tolerance and partly reduced the insulin sensitivity observed in CR vehicle mice . The metabolic differences between AL and CR vehicle mice were not accompanied by alterations in hypothalamic P14416 binding . In conclusion , blocking P14416 curtails the metabolic effects of calorie restriction . Although this suggests that the dopaminergic system could be involved in the metabolic benefits of calorie restriction , restricting access to high-fat food does not increase ( hypothalamic ) P14416 binding capacity , which argues against this inference . 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 . Binding of Y-P30 to syndecan 2/3 regulates the nuclear localization of CASK . The survival promoting peptide Y-P30 has documented neuroprotective effects as well as cell survival and neurite outgrowth promoting activity in vitro and in vivo . Previous work has shown that multimerization of the peptide with pleiotrophin ( P21246 ) and subsequent binding to syndecan ( P18827 ) -2 and -3 is involved in its neuritogenic effects . In this study we show that Y-P30 application regulates the nuclear localization of the P18827 binding partner DB01373 /calmodulin-dependent serine kinase ( CASK ) in neuronal primary cultures during development . In early development at day in vitro ( DIV ) 8 when mainly P18827 -3 is expressed supplementation of the culture medium with Y-P30 reduces nuclear CASK levels whereas it has the opposite effect at DIV 18 when P18827 -2 is the dominant isoform . In the nucleus CASK regulates gene expression via its association with the T-box transcription factor Q16650 ( Tbr-1 ) and we indeed found that gene expression of downstream targets of this complex , like the Q13224 DB01221 -receptor , exhibits a corresponding down- or up-regulation at the mRNA level . The differential effect of Y-P30 on the nuclear localization of CASK correlates with its ability to induce shedding of the ectodomain of P18827 -2 but not -3 . shRNA knockdown of P18827 -2 at DIV 18 and P18827 -3 at DIV 8 completely abolished the effect of Y-P30 supplementation on nuclear CASK levels . During early development a protein knockdown of P18827 -3 also attenuated the effect of Y-P30 on axon outgrowth . Taken together these data suggest that Y-P30 can control the nuclear localization of CASK in a P18827 -dependent manner . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . Effects of C-phycocyanin and Spirulina on salicylate-induced tinnitus , expression of DB01221 receptor and inflammatory genes . Effects of C-phycocyanin ( C-PC ) , the active component of Spirulina platensis water extract on the expressions of N-methyl D-aspartate receptor subunit 2B ( Q13224 ) , tumor necrosis factor-α ( P01375 -α ) , interleukin-1β ( IL-1β ) , and cyclooxygenase type 2 ( P35354 ) genes in the cochlea and inferior colliculus ( IC ) of mice were evaluated after tinnitus was induced by intraperitoneal injection of salicylate . The results showed that 4-day salicylate treatment ( unlike 4-day saline treatment ) caused a significant increase in Q13224 , P01375 -α , and IL-1β mRNAs expression in the cochlea and IC . On the other hand , dietary supplementation with C-PC or Spirulina platensis water extract significantly reduced the salicylate-induced tinnitus and down-regulated the mRNAs expression of Q13224 , P01375 -α , IL-1β mRNAs , and P35354 genes in the cochlea and IC of mice . The changes of protein expression levels were generally correlated with those of mRNAs expression levels in the IC for above genes . Engineering human T cells for resistance to methotrexate and mycophenolate mofetil as an in vivo cell selection strategy . Gene transfer and drug selection systems that enforce ongoing transgene expression in vitro and in vivo which are compatible with human pharmaceutical drugs are currently underdeveloped . Here , we report on the utility of incorporating human enzyme muteins that confer resistance to the lymphotoxic/immunosuppressive drugs methotrexate ( MTX ) and mycophenolate mofetil ( DB00688 ) in a multicistronic lentiviral vector for in vivo T lymphocyte selection . We found that co-expression of human dihydrofolate reductase ( P00374 (FS) ; L22F , F31S ) and inosine monophosphate dehydrogenase II ( P12268 (IY) ; T333I , S351Y ) conferred T cell resistance to the cytocidal and anti-proliferative effects of these drugs at concentrations that can be achieved clinically ( up to 0.1 µM MTX and 1.0 µM DB00603 ) . Furthermore , using a immunodeficient mouse model that supports the engraftment of central memory derived human T cells , in vivo selection studies demonstrate that huEGFRt(+) P00374 (FS+) P12268 (IY+) T cells could be enriched following adoptive transfer either by systemic administration of MTX alone ( 4.4 -fold ) , DB00688 alone ( 2.9-fold ) , or combined MTX and DB00688 ( 4.9-fold ) . These findings demonstrate the utility of both P00374 (FS)/MTX and P12268 (IY)/ DB00688 for in vivo selection of lentivirally transduced human T cells . Vectors incorporating these muteins in combination with other therapeutic transgenes may facilitate the selective engraftment of therapeutically active cells in recipients . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Increased vitamin D-driven signalling and expression of the vitamin D receptor , P35548 , and O14788 in tooth resorption in cats . Tooth resorption occurs in 20-75 % of cats ( Felis catus ) . The aetiology is not known , but vitamin D is suggested to be involved . Vitamin D acts through a nuclear receptor ( P11473 ) and increases the expression of receptor activator of nuclear factor-kappaB ligand ( rankl ) and muscle segment homeobox 2 ( msx2 ) genes . Mice lacking the muscle segment homeobox 2 ( msx2 ) gene show decreased levels of rankl , suggesting an interaction among P11473 , P35548 , and O14788 . Here , we investigated the expression of P11473 , P35548 , and O14788 proteins , and the activity of the P11473 -mediated signalling pathway ( using the quantitative polymerase chain reaction on P11473 target genes ) , in tooth resorption , and measured the serum levels of vitamin D metabolites in cats . Tooth resorption was categorized into either resorptive or reparative stages . In the resorptive stage , odontoclasts expressed P35548 and O14788 ( 100 % and 88 % , respectively ) and fibroblasts expressed P11473 and P35548 ( both at 100 % ) , whereas fibroblasts expressed O14788 in only 29 % of the sites analysed . In the reparative stage , cementoblasts expressed P11473 , P35548 , and O14788 , whereas fibroblasts expressed P11473 and P35548 , but not O14788 . The vitamin D status did not differ between the groups , based on the serum levels of DB00146 . However , increased expression of P11473 protein , and the relative gene expression levels of 1alpha-hydroxylase and the P11473 -target gene , 24-hydroxylase , indicated the involvement of an active vitamin D signalling in the pathophysiology of tooth resorption in cats . Expression and purification of growth hormone-releasing factor with the aid of dihydrofolate reductase handle . Expression of a fusion protein composed of dihydrofolate reductase and a derivative of growth hormone-releasing factor resulted in the formation of inclusion bodies in Escherichia coli at 37 degrees C . Among various chemicals , such as detergents , protein denaturants , and acetic acid , tested for the ability to dissolve the inclusion bodies , acetic acid , Brij-35 , deoxycholic acid sodium salts , guanidine-HCl , and urea showed a strong solubilizing effect without damaging the P00374 activity . DB03166 was useful in terms of preparing P01286 derivatives , since it could be easily removed by lyophilization , and this made it easy to perform the succeeding BrCN treatment for cutting out the P01286 derivative from the fusion protein . The P01286 derivative was purified by reversed phase HPLC from the BrCN digest of the acetic acid extract , and its growth hormone-releasing activity was demonstrated . However , for obtaining a highly purified fusion protein itself , solubilization of inclusion bodies by urea was preferred because urea was the only agent which did not cause serious precipitation of the regenerated fusion protein after 10-fold dilution of the extracted inclusion bodies with buffer . The fusion protein was highly purified by means of a methotrexate affinity chromatography . Oncogenes in melanoma : an update . Melanoma is a highly aggressive tumour with poor prognosis in the metastatic stage . P15056 , P01111 , and P10721 are three well-known oncogenes involved in melanoma pathogenesis . Targeting of mutated P15056 kinase has recently been shown to significantly improve overall survival of metastatic melanoma patients , underscoring the particular role of this oncogene in melanoma biology . However , recurrences regularly occur within several months , which supposedly involve further oncogenes . Moreover , oncogenic driver mutations have not been described for up to 30 % of all melanomas . In order to obtain a more complete picture of the mutational landscape of melanoma , more recent studies used high-throughput DNA sequencing technologies . A number of new oncogene candidates such as P28482 /2 , Q15303 , Q12879 , Q14832 , P63000 , and Q70Z35 were identified . Their particular role in melanoma biology is currently under investigation . Evidence for the functional relevance of some of these new oncogene candidates has been provided in in vitro and in vivo experiments . However , these findings await further validation in clinical studies . This review provides an overview on well-known melanoma oncogenes and new oncogene candidates , based on recent high-throughput sequencing studies . The list of genes discussed herein is of course not complete but highlights some of the most significant of recent findings in this area . The new candidates may support more individualized treatment approaches for metastatic melanoma patients in the future . A lactate-induced response to hypoxia . Organisms must be able to respond to low oxygen in a number of homeostatic and pathological contexts . Regulation of hypoxic responses via the hypoxia-inducible factor ( HIF ) is well established , but evidence indicates that other , HIF-independent mechanisms are also involved . Here , we report a hypoxic response that depends on the accumulation of lactate , a metabolite whose production increases in hypoxic conditions . We find that the Q9UGV2 protein is degraded in a Q9GZT9 / P40337 -dependent manner in normoxia but is protected from destruction by binding to lactate that accumulates under hypoxia . The stabilized Q9UGV2 protein binds c-Raf to mediate hypoxia-induced activation of Raf- P29323 pathway , promoting angiogenesis and cell growth . Inhibiting cellular lactate production abolishes the Q9UGV2 -mediated hypoxia responses . Our study , therefore , elucidates the molecular basis for lactate-induced hypoxia signaling , which can be exploited for the development of therapies targeting hypoxia-induced diseases . Targeting tumor necrosis factor alpha in psoriasis and psoriatic arthritis . BACKGROUND : Psoriasis is an immune-mediated chronic inflammatory disease triggered and maintained by inflammatory mediators , including P01375 . OBJECTIVE/METHODS : To summarize the role of anti- P01375 agents psoriasis therapy , focusing on the mechanisms and biological pathways involved , by reviewing relevant literature . RESULTS/CONCLUSIONS : The three P01375 antagonists currently available ( etanercept , infliximab and adalimumab ) are effective in the therapy of psoriasis and psoriatic arthritis . DB08904 and DB06674 are P01375 inhibitors not approved for therapy of psoriasis yet . In addition to neutralizing soluble P01375 , P01375 blockers bind to membrane P01375 and change the behavior of P01375 -expressing cells , resulting in hastened cell cycle arrest and apoptosis , and suppression of cytokine production . P01375 blockers may also affect adaptive immune responses by reducing T helper cell (Th)1 and Th17 responses , and favoring the development of T-regulatory cells . P01375 antagonists can regulate differentiation and activation of osteoclasts , thus reducing bone destruction in psoriatic arthritis . Anti- P01375 agents differ in their pharmacokinetics and pharmacodinamic properties , which is reflected in their therapeutic and safety profiles . The safety of P01375 antagonists has been established , and patient selection and monitoring allow risk minimization . DB00126 is dispensable for oxygen sensing in vivo . Prolyl-4-hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl-4-hydroxylase domain ( P20941 ) enzymes requires oxygen and 2-oxoglutarate as cosubstrates with iron(II) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo(-/-) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo(-/-) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum- and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF-1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl(2) . A Cys201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione . DB00074 induction in patients receiving tacrolimus-based immunosuppressive regimens . PURPOSE : The use of basiliximab induction increased significantly in recent years based on its superior efficacy and excellent safety profile demonstrated in studies with cyclosporine-based immunosuppression . However , its clinical utility in patients receiving tacrolimus-based immunosuppressive regimens is still uncertain . METHODS : We retrospectively reviewed data of 366 low immunological risk recipients of deceased donor kidney transplants . Of them , 134 received basiliximab and tacrolimus ( TAC- P01589 ) , 100 received basiliximab and delayed tacrolimus(dTAC- P01589 ) , and 132 patients received tacrolimus without basiliximab(TAC-No) . The endpoints were the incidence of acute rejection , graft function , and patient and graft survivals at 1 year . RESULTS : The incidence of acute rejection was higher in dTAC- P01589 compared to TAC-IL-2RA and TAC-No Groups ( 33 vs.14.9 vs. 14.3 % , p < 0.001 ) . Inferior creatinine clearance was observed in dTAC- P01589 Group compared to TAC- P01589 and TAC-No Groups at months 1 ( 41.6 vs. 49.9 vs. 44.8 mL/min , p = 0.004 ) , 3 ( 49.8 vs. 57.2 vs. 53.5 mL/min , p = 0.017 ) , and 6 ( 53.1 vs. 61.8 vs. 57.0 mL/min , p = 0.001 ) . Patients who received basiliximab ( TAC- P01589 and dTAC- P01589 Groups ) had lower incidence of posttransplant diabetes ( 24 vs.18 vs. 39.3 % , p = 0.009 ) . Patient and graft survivals were similar among the groups . CONCLUSIONS : In low immunological risk kidney transplant recipients receiving tacrolimus , the use of basiliximab induction was not associated with lower rejection rates and did not allow delayed tacrolimus introduction . Phase II study of weekly intravenous oxaliplatin combined with oral daily capecitabine and radiotherapy with biologic correlates in neoadjuvant treatment of rectal adenocarcinoma . PURPOSE : To evaluate the efficacy of a combination of capecitabine , oxaliplatin , and radiotherapy ( RT ) in the neoadjuvant treatment of Stage II and III rectal cancers . METHODS : DB01101 was given at 725 mg/m(2) orally twice daily Monday through Friday concurrently with RT . DB00526 was given intravenously at 50 mg/m(2) once weekly five times starting the first day of RT . The radiation dose was 50.4 Gy in 28 fractions ( 1.8 Gy/fraction ) , five fractions weekly . Endorectal tumor biopsies were obtained before treatment and on the third day of treatment to explore the effects of treatment on thymidine phosphorylase , thymidylate synthase , excision repair cross-complementing rodent repair deficiency complementation group 1 ( P07992 ) , and apoptosis . RESULTS : A total of 25 patients were enrolled in this study ; 6 patients ( 24 % ) had a complete pathologic response . T-downstaging occurred in 52 % of patients , and N-downstaging occurred in 53 % . Grade 3 diarrhea was the most common Grade 3-4 toxicity , occurring in 20 % of patients . Only 2 patients experienced disease recurrence , with a median of 20 months of follow-up . P04818 , thymidine phosphorylase , P07992 , and apoptosis did not vary significantly between the pretreatment and Day 3 tumor biopsies , nor did they predict for T-downstaging or a complete pathologic response . CONCLUSION : DB01101 at 725 mg/m(2) orally twice daily , oxaliplatin 50 mg/m(2)/wk , and RT at 50.4 Gy is an effective neoadjuvant combination for Stage II and III rectal cancer and results in a greater rate of complete pathologic responses than historically shown in fluoropyrimidine plus RT controls . Parathyroid-hormone-related protein induces expression of receptor activator of NF-{kappa}B ligand in human periodontal ligament cells via a DB02527 /protein kinase A-independent pathway . Periodontal ligament ( PDL ) cells play important roles in root resorption of human deciduous teeth by odontoclasts ( osteoclast-like cells ) . However , it is unclear how PDL cells regulate osteoclastogenesis . We examined the effects of P12272 , TGF-beta , and P01133 , which are all secreted by the tooth germ , on tartrate-resistant acid-phosphatase-positive ( TRAP+ ) cell formation using co-cultures of human PDL cells and mouse spleen cells . Only P12272 promoted TRAP+ cell formation in co-cultures . P12272 induced receptor activator of NF-kappaB ligand ( O14788 ) mRNA expression and slightly reduced osteoprotegerin ( O00300 ) expression in PDL cells . The DB02527 /PKA inhibitors Rp- DB02527 , H89 , and PKI did not affect P12272 -induced TRAP+ cell formation . The PKC inhibitor , Ro-32-0432 , suppressed O14788 expression in PDL cells and P12272 -induced TRAP+ cell formation . However , this inhibitor directly modulated the number of osteoclast precursors . Thus , P12272 induces osteoclastogenesis by increasing the relative expression level of O14788 vs. O00300 in PDL cells via a DB02527 /PKA-independent pathway . ABBREVIATIONS : P12272 , parathyroid-hormone-related protein ; TGF-beta , transforming growth factor-beta ; P01133 , epidermal growth factor ; O14788 , receptor activator of NF-kappaB ligand ; O00300 , osteoprotegerin ; PDL , periodontal ligament ; TRAP , tartrate-resistant acid phosphatase ; PKA , protein kinase A ; PKC , protein kinase C ; Q96HU1 , mitogen-activated protein ; P29323 , extracellular signal-regulated kinase ; DB02527 , cyclic DB00640 3'5'-Monophosphate . High mobility group box protein-1 promotes cerebral edema after traumatic brain injury via activation of toll-like receptor 4 . Traumatic brain injury ( TBI ) is a major cause of mortality and morbidity worldwide . Cerebral edema , a life-threatening medical complication , contributes to elevated intracranial pressure ( ICP ) and a poor clinical prognosis after TBI . Unfortunately , treatment options to reduce post-traumatic edema remain suboptimal , due in part , to a dearth of viable therapeutic targets . Herein , we tested the hypothesis that cerebral innate immune responses contribute to edema development after TBI . Our results demonstrate that high-mobility group box protein 1 ( P09429 ) was released from necrotic neurons via a Q13224 -mediated mechanism . P09429 was clinically associated with elevated ICP in patients and functionally promoted cerebral edema after TBI in mice . The detrimental effects of P09429 were mediated , at least in part , via activation of microglial toll-like receptor 4 ( O00206 ) and the subsequent expression of the astrocytic water channel , aquaporin-4 ( P55087 ) . Genetic or pharmacological ( VGX-1027 ) O00206 inhibition attenuated the neuroinflammatory response and limited post-traumatic edema with a delayed , clinically implementable therapeutic window . Human and rodent tissue culture studies further defined the cellular mechanisms demonstrating neuronal P09429 initiates the microglial release of interleukin-6 ( P05231 ) in a O00206 dependent mechanism . In turn , microglial P05231 increased the astrocytic expression of P55087 . Taken together , these data implicate microglia as key mediators of post-traumatic brain edema and suggest P09429 - O00206 signaling promotes neurovascular dysfunction after TBI . 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 . Evaluation of a microarray for genotyping polymorphisms related to xenobiotic metabolism and DNA repair . We present an oligonucleotide microarray ( " MetaboChip " ) based on the arrayed primer extension ( P27695 ) technique , allowing genotyping of single nucleotide polymorphisms ( SNPs ) in genes of interest for cancer susceptibility and pharmacogenetics . P27695 consists of a sequencing reaction primed by an oligonucleotide anchored with its 5' end to a glass slide and terminating one nucleotide before the polymorphic site . The extension with one fluorescently labeled dideoxynucleotide complementary to the template reveals the polymorphism . Ninety-three SNPs in 42 genes were selected among those resequenced in the context of the SNP500 project , using a set of 102 reference DNA samples from the Coriell Biorepository . Selected SNPs belong to the following genes : P00325 , P05091 , P27695 , CDKN2A , P21964 , P04798 , P05177 , Q16678 , P11509 , P33261 , P11712 , P05181 , P08684 , P14416 , P21917 , P07099 , P07992 , P18074 , Q92889 , P28715 , P30550 , O15217 , P21266 , P09211 , GSTT2 , LIG3 , Q00987 , P16455 , P05164 , NAT1 , NAT2 , P15559 , O15527 , P12004 , P06746 , Q01959 , P04179 , P04637 , P18887 , O43543 , O43542 , and O15287 . We assessed the performance of P27695 by comparing the results obtained with MetaboChip against those reported by the SNP500 . Among 88 SNPs that yielded signals , 6 showed less than 99 % of concordance , whereas 82 performed accurately , showing that P27695 is a reliable and sensitive genotyping method . 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 . The receptor tyrosine kinase inhibitor amuvatinib ( DB05216 ) sensitizes tumor cells to radio- and chemo-therapies in part by inhibiting homologous recombination . BACKGROUND AND PURPOSE : Q06609 is a key protein involved in homologous recombination ( HR ) and a potential target for radiation- and chemotherapies . Amuvatinib ( formerly known as DB05216 ) is a novel receptor tyrosine kinase inhibitor that targets c- P10721 and PDGFRα and can sensitize tumor cells to ionizing radiation ( IR ) . Here , we studied amuvatinib mechanism on Q06609 and functional HR . MATERIALS AND METHODS : Protein and RNA analyses , direct repeat green fluorescent protein ( DR-GFP ) assay and polysomal fractioning were used to measure HR efficiency and global translation in amuvatinib-treated H1299 lung carcinoma cells . Synergy of amuvatinib with IR or mitomycin c ( DB00305 ) was assessed by clonogenic survival assay . RESULTS : Amuvaninib inhibited Q06609 protein expression and HR . This was associated with reduced ribosomal protein S6 phosphorylation and inhibition of global translation . Amuvatinib sensitized cells to IR and DB00305 , agents that are selectively toxic to HR-deficient cells . CONCLUSIONS : Amuvatinib is a promising agent that may be used to decrease tumor cell resistance . Our work suggests that this is associated with decreased Q06609 expression and function and supports the further study of amuvatinib in combination with chemotherapy and radiotherapy . Imbalance of the mitochondrial pro- and anti-apoptotic mediators in neuroblastoma tumours with unfavourable biology . It has been proposed that a lack of apoptosis plays an important role in neuroblastoma ( NB ) progression . We therefore screened cDNA array filters , including 198 apoptotic genes , in order to identify mRNA transcripts that are differentially expressed in tumours with unfavourable versus favourable biology . Twenty-one genes were analysed further using real-time reverse-transcriptase-polymerase chain reaction ( RT-PCR ) . Significantly lower levels of P63167 ( P63167 ; P(c)(corrected) = 0.0054 ) and P04629 ( TrkA ; P(c) = 0.039 ) were found in NB tumours with unfavourable biology . In addition , P55957 , P10415 , O14727 , P42575 , P42574 and P55211 were found to be preferentially expressed in tumours with favourable biology , whereas P38936 ( P38936 ) , P01589 , and Q07820 , were found to be preferentially expressed in NB tumours with unfavourable biology . In conclusion , mRNA levels of transcripts encoding pro-apoptotic mediators of the mitochondrial apoptotic pathway were found to be expressed to a lower extent in tumours with unfavourable biology . Our data also suggest that the mitochondrial pathway is suppressed in advanced stages of NB tumours , due to an imbalance between anti-apoptotic and pro-apoptotic mediators which is a finding that may have therapeutic significance .

MH_train_1538

MH_train_1538

MH_train_1538

interacts_with DB06779?

multiple_choice

P01308 up-regulates vascular endothelial growth factor and stabilizes its messengers in endometrial adenocarcinoma cells . Angiogenesis is crucial for tumor growth and dissemination . Vascular endothelial growth factor ( P15692 ) is a potent angiogenic factor that promotes vascular growth and therefore tumoral growth and metastasis . Overweight , frequently associated with hyperinsulinemia , constitutes the major risk factor for endometrial carcinoma . Thus , elevated insulin levels may partly explain the increased risk of endometrial cancer observed in obese postmenopausal women . The aim of the present work was to test the role of insulin in the control of P15692 expression in endometrial carcinoma cells ( O14777 -1A ) . We have shown that insulin induced a biphasic expression of P15692 messenger ribonucleic acid , with an early , but low , induction ( 4 h of stimulation ) and a delayed , but high , induction ( 24 h ) . The delayed effect of insulin on P15692 expression involved transcriptional and posttranscriptional regulation , as evidenced by the increased rate of P15692 transcription and the prolonged half-life of P15692 messenger ribonucleic acid . Simultaneously we observed higher levels of P15692 protein in the conditioned medium of stimulated cells compared with unstimulated ones . Therefore , insulin could contribute to the increased risk of endometrial carcinoma due to its ability to induce P15692 expression and thus participate in the maintenance of an angiogenic phenotype . APELIN promotes hematopoiesis from human embryonic stem cells . Transcriptional profiling of differentiating human embryonic stem cells ( hESCs ) revealed that Q9H2W2 -positive mesodermal precursors were enriched for transcripts encoding the G-protein-coupled APELIN receptor ( P35414 ) . P35414 -positive cells , identified by binding of the fluoresceinated peptide ligand , APELIN ( Q9ULZ1 ) , or an anti- P35414 mAb , were found in both posterior mesoderm and anterior mesendoderm populations and were enriched in hemangioblast colony-forming cells ( Bl- Q15814 ) . The addition of Q9ULZ1 peptide to the media enhanced the growth of embryoid bodies ( EBs ) , increased the expression of hematoendothelial genes in differentiating hESCs , and increased the frequency of Bl-CFCs by up to 10-fold . Furthermore , Q9ULZ1 peptide also synergized with P15692 to promote the growth of hESC-derived endothelial cells . These studies identified Q9ULZ1 as a novel growth factor for hESC-derived hematopoietic and endothelial cells . [ Relationship between the protein expression of P04637 , c-erbB-2 , vascular endothelial growth factor and P16070 and the survival rates of stage II ( colorectal cancer patients without radiochemotherapy after radical resection ] . OBJECTIVE : To investigate the relation between protein expression of 4 genes [ P04637 ,c-erbB-2,vascular endothelial factors( P15692 ) and P16070 ] and survival rate in stage II ( colorectal cancer(CRC) patients without radiochemotherapy after radical resection . METHODS : One hundred and fifty-nine cases of stage II ( CRC without radiochemotherapy were enrolled in this study . The clinicopathological date and 5-year follow-up data were reviewed . Streptavidin-peroxidase immunohistochemical technique was used to detect the expression of P04637 , c-erbB-2 , P15692 and P16070 in formalin-fixed , paraffin embedded sections of CRC tissues from above 159 patients . RESULTS : The 5-year survival rate was 82.4 % . The rates of positive expression of P04637 , c-erbB-2 , P15692 and P16070 were 58.5 % ( 93/159 ) , 26.4 % ( 42/159 ) , 57.9 % ( 92/159 ) and 40.0 % ( 54/159 ) respectively . The 5-year survival rates of positive expression patients were not significantly different with those of negative expression . chi(2) analysis showed that the positive expressions of 4 genes had no relationships with the prognosis . CONCLUSION : The expression of 4 gene proteins has no relationship with the prognosis of stage II ( CRC patients without radiochemotherapy after radical resection . P15559 as a target enzyme for biochemical modulation of mitomycin C . We studied a selective enhancement of the mitomycin C ( DB00305 ) -induced antitumor effect focusing on the intracellular metabolism by NAD(P)H:quinone oxidoreductase ( P15559 , DTD ) . The level of cellular DTD activity related well to the degree of DB00305 -induced DNA total cross links and cell growth inhibition in human cancer cell lines , KB , PH101 , SH101 and K562 . A DTD inhibitor , dicoumarol ( DIC ) or flavin adenine dinucleotide ( DB03147 ) , inhibited the DB00305 -induced DNA damage and cytotoxicity at a non-toxic concentration . The DTD-mediated DB00305 activation was pH-dependent , and highest at pH 6 and lowest at pH 8 . Although an inverse relationship appeared to exist between DTD activity and DB00305 efficacy in human xenografts implanted into nude mice and 9 fresh human tumor specimens , the investigation in 3 culture cells , O14777 -46 , HCC-48 and HCC-50 , established from those xenografts , showed that DTD activated DB00305 in a pH-dependent manner as well as the other cell lines . Significant tumor pH reduction from 7.1 to 6.7 by continuous glucose infusion also increased the DB00305 -induced tumor growth inhibition in the human tumor xenografts . Thus , we conclude that bioreductive activation by DTD in a pH-dependent manner may be of key importance in the DB00305 -induced antitumor effect and that an increased DB00305 efficacy at a reduced pH caused by hyperglycemia may be applied to clinical use as a new manipulation for a biochemical modulation of DB00305 . 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 . Isoeugenodilol inhibits smooth muscle cell proliferation and neointimal thickening after balloon injury via inactivation of P27361 /2 pathway . The purpose of this study was to determine the efficacy and the possible mechanism of action of the synthesized drug isoeugenodilol ( a new third-generation beta-adrenoceptor blocker ) on the growth factor-induced proliferation of cultured rat vascular smooth muscle cells ( VSMCs ) and neointimal formation in a rat carotid arterial balloon injury model . Isoeugenodilol significantly inhibited 10 % FBS , 20 ng/ml DB00102 , and 20 ng/ml vascular endothelial growth factor ( P15692 ) -induced proliferation . In accordance with these findings , isoeugenodilol revealed blocking of the FBS-inducible progression through the G(0)/G(1) to the S phase of the cell cycle in synchronized cells . Neointimal formation , measured 14 days after injury , was reduced by the oral administration of isoeugenodilol ( 10 mg/kg/day ) . In an in vitro assay , isoeugenodilol inhibited the migration of VSMCs stimulated by DB00102 . These findings indicate that isoeugenodilol shows an inhibitory potency on neointimal formation due to inhibition of both migration and proliferation of VSMCs . In addition , isoeugenodilol in concentration-dependent manner decreased the levels of phosphorylated P27361 /2 in both VSMCs and balloon-injured carotid arteries . The levels of phosphorylated Q02750 /2 and Pyk2 as well as intracellular Ca(2+) and reactive oxygen species ( ROS ) were in concentration-dependent manner reduced by isoeugenodilol . Taken together , these results indicate that isoeugenodilol may suppress mitogen-stimulated proliferation and migration partially through inhibiting cellular ROS and calcium , and hence , through activation of the Pyk2- P27361 /2 signal pathway . This suggests that isoeugenodilol has potential for the prevention of atherosclerosis and restenosis . Emerging biologic drugs for the treatment of rheumatoid arthritis . This article reviews the role of emerging biologic drugs for the treatment of rheumatoid arthritis ( RA ) . Besides anti-tumor necrosis factor ( P01375 ) -alpha and anti-interleukin ( IL ) -1 agents ( DB00065 , DB00051 , DB00005 and DB00026 ) whose clinical efficacy is now established , new drugs have been proposed for the therapy of rheumatoid arthritis patients not responding to conventional treatments . These approaches include the blockade of B-cell activity with anti- P11836 monoclonal antibody ( DB00073 ) and the inhibition of T-cell activation with fusion protein DB01281 . Moreover , promising results have been obtained in animal models utilizing suppressors of cytokine signaling ( Q9NSE2 ) and dominant-negative P01375 variants to inactivate P01375 signaling . 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 . Altered growth factor expression in the aging penis : the Brown-Norway rat model . The objective of the present study was to evaluate age-related changes in the protein and gene expression of modulators of erectile function ( nitric oxide [ NO ] and endothelin-1 [ ET-1 ] ) and growth factors such as transforming growth factor ( TGF-beta1 ) and vascular endothelial growth factor ( P15692 ) in the penile tissue of Brown-Norway ( BN ) rats . Young and old BN male rats were euthanized , and the penile tissue was processed for immunohistochemical and molecular analyses . Total RNA was extracted , and an Access reverse transcription-polymerase chain reaction ( RT-PCR ) system was used for messenger RNA ( mRNA ) expression analysis . Immunohistochemical studies showed a decreased expression of endothelial nitric oxide synthase ( P29474 ) protein and an increased staining for ET-1 . Quantitative analysis of PCR products revealed decreased levels of P15692 mRNA expression in the old population of rats . The most significant decrease was detected between bands corresponding to splice forms 164 ( 21 % ) and 120 ( 18 % ) . The observed alterations in the gene expression of growth factors such as P15692 may contribute to the abnormal age-related morphological and physiological alterations in the erectile tissue . 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 . 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 . Absolute bioavailability and effect of formulation change , food , or elevated pH with rabeprazole on cobimetinib absorption in healthy subjects . DB05239 is a potent and highly selective inhibitor of Q02750 /2 . Since cobimetinib exhibited absorption variability in cancer patients , a series of single-dose studies in healthy subjects were conducted to determine absolute bioavailability and elucidate potential effects of formulation , food , and elevated gastric pH on cobimetinib bioavailability . Three crossover trials were performed with a 20 mg cobimetinib oral dose : absolute bioavailability using a 2 mg intravenous infusion ( n = 13 ) , relative bioavailability of tablets versus capsules and food effect ( n = 20 ) , and drug interaction with a proton pump inhibitor ( 20 mg of rabeprazole daily for 5 days prior to cobimetinib administration ; n = 20 ) . Absolute bioavailability of cobimetinib was 46.2 % ( 24.2 , CV % ) , likely due to metabolism rather than incomplete absorption . The mean systemic clearance of cobimetinib was low ( 11.7 L/h [ 28.2 , CV % ] ) . Administration of cobimetinib tablets with a high-fat meal delayed drug absorption ( prolonged tmax ) but had no statistically significant effect on cobimetinib exposure ( Cmax and AUC0-∞ ) . Tablet and capsule formulations of cobimetinib showed comparable exposures . DB05239 exhibited delayed absorption ( tmax ) in the presence of rabeprazole , with no statistically significant effects on drug exposure ( Cmax and AUC0-∞ ) in the fasted state . In conclusion , cobimetinib oral absorption was not affected by change in formulation , food , or elevated gastric pH . Selectin ligand expression regulates the initial vascular interactions of colon carcinoma cells : the roles of CD44v and alternative sialofucosylated selectin ligands . Selectin-mediated binding of tumor cells to platelets , leukocytes , and vascular endothelium may regulate their hematogenous spread in the microvasculature . We recently reported that P16070 variant isoforms ( CD44v ) on LS174T colon carcinoma cells possess selectin binding activity . Here we extended those findings by showing that T84 and Colo205 colon carcinoma cells bind selectins via sialidase-sensitive O-linked glycans presented on CD44v , independent of heparan and chondroitin sulfate . To assess the functional role of CD44v in selectin-mediated binding , we quantified the adhesion to selectins of T84 cell subpopulations sorted based on their P16070 expression levels and stable LS174T cell lines generated using P16070 short hairpin RNA . High versus low P16070 -expressing T84 cells tethered more efficiently to P- and P14151 , but not P16581 , and rolled more slowly on P- and P16581 . Knocking down P16070 expression on LS174T cells inhibited binding to P16109 and increased rolling velocities over P- and P14151 relative to control-transfected cells , without affecting tethering and rolling on P16581 , however . Blot rolling analysis revealed the presence of alternative sialylated glycoproteins with molecular masses of approximately 170 and approximately 130 kDa , which can mediate selectin binding in P16070 -knockdown cells . DB01109 diminishes the avidity of colon carcinoma cells for P- and P14151 , which may compromise integrin-mediated firm adhesion to host cells and mitigate metastasis . Our finding that CD44v is a functional P16109 ligand on colon carcinoma provides a novel perspective on the enhanced metastatic potential associated with tumor CD44v overexpression and the role of selectins in metastasis . [ Intravitreal anti- P15692 therapy with bevacizumab for neovascular AMD ] . PURPOSE : To report on the efficacy of intravitreal bevacizumab as off-label therapy in different angiographic subtypes in neovascular age-related macular degeneration ( AMD ) . METHODS : Seventy-five patients with neovascular AMD and recent disease progression were classified into different angiographic subtypes and were treated with intravitreal bevacizumab ( 1.25 mg/0.05 ml ) at 6-week intervals . Patients with subfoveal classic choroidal neovascularization ( CNV ) also received photodynamic therapy . ETDRS visual acuity , ophthalmic exams , and optic coherence tomography ( O75051 ) were performed before treatment , 1 week after treatment , and then on a 6-week basis . DB00693 angiographies and medical check-ups were also done . RESULTS : DB00112 led to stabilization of visual acuity ( loss of less than 15 letters ) in all angiographic subtypes during a follow-up of 37+/-13 weeks . Patients with occult extrafoveal CNV ( n=6 ) profited the most and gained 2+/-2 lines . Treatment with intravitreal bevacizumab was very well tolerated in all patients , with neither systemic nor intraocular side effects , with the exception of one retinal pigment epithelium tear . CONCLUSION : Intravitreal bevacizumab treatment is efficacious in all angiographic CNV subtypes and leads to reduction of macular edema and stabilization or improvement in visual acuity . 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 . Proteomic discovery of genistein action in the rat mammary gland . DB01645 , the primary isoflavone component of soy , consumed in diet during the prepubertal period suppresses chemically induced mammary cancer in rats . The current study used two-dimensional gel electrophoresis ( 2-DE ) /MS-based proteomic technology to identify proteins responsible for genistein breast cancer protection In Vivo . Female offspring were exposed via lactating dams treated with 250 mg genistein/kg AIN-76A diet from days 1 to 21 postpartum ( prepubertal period ) . Mammary glands were collected at 21 and 50 day of age and subjected to 2-DE/MS and immuno-blot analyses . Twenty-three proteins were determined to be differentially regulated ( p < 0.05 ) and identified using 2-DE , followed by MALDI-TOF/TOF or LC- P19957 -MS/MS . Five of these proteins were validated by immuno-blots . P07355 was significantly increased at 21 days yet found to be decreased at 50 days . Fetuin B was found to be unchanged at day 21 but increased at day 50 . P00558 ( P00558 ) was unchanged at day 21 but decreased at day 50 . P06396 was increased at day 21 but not at day 50 . P30101 ( P30101 ) was decreased at day 21 and unchanged at day 50 . Also , we found that vascular endothelial growth factor receptor 2 ( P15692 -R2 ) and epidermal growth factor receptor ( P01133 -R ) were decreased in mammary glands of 50-day-old rats treated prepubertally with genistein . This study demonstrates the usefulness of proteomics for the discovery of key proteins involved in signaling pathways to understand genistein mechanisms of action in breast cancer prevention . 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 . Caspase activation in etoposide-treated fibroblasts is correlated to P29323 phosphorylation and both events are blocked by polyamine depletion . Activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 is correlated to cell survival , but in some cases ERKs can act in signal transduction pathways leading to apoptosis . Treatment of mouse fibroblasts with 20 microM etoposide elicited a sustained phosphorylation of P29323 1/2 , that increased until 24 h from the treatment in parallel with caspase activity . The inhibitor of P29323 activation PD98059 abolished caspase activation , but caspase inhibition did not reduce P29323 1/2 phosphorylation , suggesting that P29323 activation is placed upstream of caspases . Both P29323 and caspase activation were blocked in cells depleted of polyamines by the ornithine decarboxylase inhibitor alpha-difluoromethylornithine ( DB06243 ) . In etoposide-treated cells , DB06243 also abolished phosphorylation of c-Jun NH(2)-terminal kinases triggered by the drug . Polyamine replenishment with exogenous putrescine restored the ability of the cells to undergo caspase activation and P29323 1/2 phosphorylation in response to etoposide . P11926 activity decreased after etoposide , indicating that DB06243 exerts its effect by depleting cellular polyamines before induction of apoptosis . These results reveal a role for polyamines in the transduction of the death signal triggered by etoposide .

MH_train_1539

MH_train_1539

MH_train_1539

interacts_with DB01114?

multiple_choice

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 . 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 . Modulation of diabetes with gonadotropin-releasing hormone antagonists in the nonobese mouse model of autoimmune diabetes . The nonobese mouse model of autoimmune diabetes ( NOD mouse ) exhibits a strain-dependent preponderance of disease in females . Castration of male NOD mice leads to an increased incidence of diabetes , suggesting that testosterone directly modulates the expression of diabetes in the NOD mouse . However , castration also modulates hypothalamic and pituitary hormone production via removal of the negative feedback effects of testosterone . One hypothalamic hormone with immunomodulatory properties whose expression is increased by castration is DB00644 . To test whether the increased incidence of diabetes in castrated male NOD mice is related to an increase in DB00644 activity , we treated castrated male NOD mice with Antide , a P30968 antagonist , to determine the effect on the incidence and timing of onset of diabetes . The prevalence of diabetes at 40 wk of age in male NOD mice was 50 % in sham-operated mice , compared with an 83 % prevalence in castrated males . Antide administration prevented the increased incidence of diabetes in the castrated male mice . Antide reduced total serum IgG levels , P05231 cytokine expression in cultured splenocytes , and the lymphocytic infiltration of islets . DB00644 administration exerted reciprocal effects , leading to earlier timing of onset of diabetes and increases in serum total IgG levels . We conclude that DB00644 modulates the expression of diabetes in the NOD mouse independently of gonadal steroids . 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 . 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 . Hormonal regulation of human trophoblast differentiation : a possible role for 17beta-estradiol and DB00644 . We have examined the role of 17beta-estradiol and gonadotropin releasing hormone ( DB00644 ) in the regulation of functional differentiation in human trophoblasts . In contrast to its recognized functions as a proliferation-promoting hormone in a variety of cell types , we found that 17beta-estradiol induced terminal differentiation in human trophoblastic cells , and that this event was estrogen-receptor-mediated . This process involved a loss in expression of Cyclins A2 and E , and a coincident increase in p27(Kip1) . The anti-proliferative effects of 17beta-estradiol were annulled by specific transforming growth factor-beta 1 ( TGFbeta1 ) -neutralizing antibody , suggesting that 17beta-estradiol may mediate its growth-inhibitory actions , through TGFbeta1 activity . Following exposure to DB06719 , cultured human trophoblastic cells stopped proliferating and formed functionally mature syncytiotrophoblasts . This differentiation event , that involved a drastic loss in expression of proliferating-cell-nuclear-antigen , could be blocked by DB00050 , suggesting the involvement of functional DB00644 receptors . Preliminary studies on the characterization of the human placental P30968 , indicate the presence of multiple receptor isoforms across human gestation . 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 . High biochemical selectivity of tadalafil , sildenafil and vardenafil for human phosphodiesterase 5A1 ( O76074 ) over PDE11A4 suggests the absence of PDE11A4 cross-reaction in patients . The physiological role of phosphodiesterase (PDE)11 is unknown and its biochemical characteristics are poorly understood . We have expressed human DB00117 -tagged PDE11A4 and purified the enzyme to apparent homogeneity . PDE11A4 displays K(m) values of 0.97 microM for cGMP and 2.4 microM for DB02527 , and maximal velocities were 4- to 10-fold higher for DB02527 than for cGMP . Given the homology between PDE11 and O76074 , we have compared the biochemical potencies of tadalafil ( DB00820 , Lilly- Q9Y6W8 ) , vardenafil ( DB00862 , Bayer-GSK ) , and sildenafil ( Viagra , Pfizer Inc. ) for PDE11A4 and PDE5A1 . PDE5A1/PDE11A4 selectivities are 40- , 9300- , and 1000-fold for tadalafil , vardenafil , and sildenafil , respectively . This suggests that none of these three compounds is likely to crossreact with PDE11A4 in patients . Establishment of pemetrexed-resistant non-small cell lung cancer cell lines . DB00642 ( P15941 ) , a multitargeted antifolate with manageable toxicity , is active against non-squamous non-small cell lung cancer ; however , most patients eventually acquire resistance to P15941 . To elucidate the resistant mechanism , we established P15941 -resistant lung adenocarcinoma cell lines . Two parental cell lines , PC-9 and A549 , were treated with step-wise increasing concentrations of P15941 . Growth inhibition was determined by the 3-[4,5-dimethyl-thizol-2-yl]-2,5-diphenyltetrazolium bromide assay . Expression of the genes encoding thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyltransferase ( GARFT ) was analyzed by quantitative real-time reverse transcriptase polymerase chain reaction . The four PC-9 sublines were more resistant than the PC-9 cell line to P15941 ( 2.2- , 2.9- , 8.4- , and 14.3-fold , respectively ) . The four A549 sublines also showed more resistance to P15941 ( 7.8- , 9.6- , 42.3- , and 42.4-fold , respectively ) than the parent cell line . All resistant sublines showed cross-resistance to cisplatin , but not to docetaxel , vinorelbine , 5-fluorouracil , or the active metabolite of irinotecan , SN-38 . All P15941 -resistant sublines expressed more TS than the parental cells , by polymerase chain reaction and Western blotting . P00374 was significantly increased in the four P15941 -resistant A549 sublines . GARFT did not correlate with resistance to P15941 . In summary , P15941 -resistant cells remained sensitive to docetaxel , vinorelbine , 5-fluorouracil , and irinotecan . TS expression appeared to be associated with resistance to P15941 . Effects of inhaled prostacyclin analogue on chronic hypoxic pulmonary hypertension . Inhaled DB01240 has been reported to elicit pulmonary vasodilation , but whether it is also effective in treating chronic hypoxic pulmonary hypertension is still uncertain . We designed this study to address the in vivo effectiveness of inhaled DB05229 , a stable DB01240 analogue , on pulmonary vascular tone during hypoxic exposure in normoxic ( N ) and chronically hypoxic ( CH ) rats . Pulmonary vasodilation was observed by low-dose inhaled DB05229 in N rats , but not in CH rats . It was not until higher doses of DB05229 were given that pulmonary vasodilation was obtained in CH rats . When the agent was continuously administered by an intravascular route at the inhaled dose , it elicited no vasodilation in N rats . On the contrary , it elicited profound vasodilation in CH rats , although a concomitant systemic hypotension was observed . The P43119 mRNA expression was unchanged in the lungs of CH rats compared with that of N rats . We conclude that low doses of aerosolized DB05229 may reduce pulmonary vascular tone in rats without preexisting lung diseases . In contrast , when hypoxic pulmonary hypertension is present , the threshold of DB05229 inhalation was elevated to provoke pulmonary vasodilation . Murine lupus susceptibility locus Sle1a controls regulatory T cell number and function through multiple mechanisms . The Sle1 locus is a key determinant of lupus susceptibility in the NZM2410 mouse model . Within Sle1 , we have previously shown that Sle1a expression enhances activation levels and effector functions of P01730 (+) T cells and reduces the size of the P01730 (+)CD25(+)Foxp3(+) regulatory T cell subset , leading to the production of autoreactive T cells that provide help to chromatin-specific B cells . In this study , we show that Sle1a P01730 (+) T cells express high levels of Q9Y6W8 , which is consistent with their increased ability to help autoreactive B cells . Furthermore , Sle1a P01730 (+)CD25(+) T cells express low levels of Foxp3 . Mixed bone marrow chimeras demonstrated that these phenotypes require Sle1a to be expressed in the affected P01730 (+) T cells . Expression of other markers generally associated with regulatory T cells ( Tregs ) was similar regardless of Sle1a expression in Foxp3(+) cells . This result , along with in vitro and in vivo suppression studies , suggests that Sle1a controls the number of Tregs rather than their function on a per cell basis . Both in vitro and in vivo suppression assays also showed that Sle1a expression induced effector T cells to be resistant to Treg suppression , as well as dendritic cells to overproduce P05231 , which inhibits Treg suppression . Overall , these results show that Sle1a controls both Treg number and function by multiple mechanisms , directly on the Tregs themselves and indirectly through the response of effector T cells and the regulatory role of dendritic cells . Behind the curtain : cellular mechanisms for allosteric modulation of calcium-sensing receptors . DB01373 -sensing receptors ( P41180 ) are integral to regulation of systemic Ca(2+) homeostasis . Altered expression levels or mutations in P41180 cause Ca(2+) handling diseases . P41180 is regulated by both endogenous allosteric modulators and allosteric drugs , including the first Food and Drug Administration-approved allosteric agonist , DB01012 HCl ( Sensipar® ) . Recent studies suggest that allosteric modulators not only alter function of plasma membrane-localized P41180 , but regulate P41180 stability at the endoplasmic reticulum . This brief review summarizes our current understanding of the role of membrane-permeant allosteric agonists in cotranslational stabilization of P41180 , and highlights additional , indirect , signalling-dependent role(s) for membrane-impermeant allosteric drugs . Overall , these studies suggest that allosteric drugs act at multiple cellular organelles to control receptor abundance and hence function , and that drug hydrophobicity can bias the relative contributions of plasma membrane and intracellular organelles to P41180 abundance and signalling . P35354 inhibitors ameliorate experimental autoimmune encephalomyelitis through modulating P01579 and P22301 production by inhibiting T-bet expression . The P35354 inhibitors DB00533 ( Rof ) and DB01283 ( Lum ) were evaluated in experimental autoimmune encephalomyelitis ( EAE ) , the model of multiple sclerosis ( MS ) . Administration of Rof and Lum significantly reduced the incidence and severity of EAE , which was associated with the inhibition of Q16653 35-55 lymphocyte recall response , anti- Q16653 35-55 T cell responses , and modulation of cytokines production . In vitro Rof and Lum inhibited primary T cells proliferation and modulated cytokine production . These findings highlight the fact that Rof and Lum likely prevents EAE by modulating Th1/Th2 response , and suggest its utility in the treatment of MS and other autoimmune diseases . P06401 modulator DB05366 down-regulates vascular endothelial growth factor , adrenomedullin and their receptors and modulates progesterone receptor content in cultured human uterine leiomyoma cells . BACKGROUND : This study was conducted to evaluate the effects of graded concentrations ( 10(-8) , 10(-7) and 10(-6) M ) of progesterone receptor ( PR ) modulator DB05366 on the protein contents of PR , of vascular endothelial growth factor ( P15692 ) , adrenomedullin ( P35318 ) and their receptors in cultured human uterine leiomyoma and matching myometrial cells . METHODS : PR-A , PR-B , P15692 , P49765 , P15692 receptor ( VEGFR ) -1 , P35968 , P35318 and P35318 receptor ( O15218 ) contents were assessed by Western blot analysis . RESULTS : Treatment with 100 ng/ml progesterone increased P15692 , P49765 and P35318 contents in cultured leiomyoma cells and normal myometrial cells . The concomitant treatment with 10(-6) M DB05366 significantly decreased the progesterone-induced P15692 , P49765 and P35318 contents in cultured leiomyoma cells but not in normal myometrial cells . DB05366 treatment alone decreased P17948 , P35968 and O15218 contents in cultured leiomyoma cells but not in normal myometrial cells . DB05366 treatment increased PR-A and decreased PR-B contents in cultured leiomyoma cells in a dose-dependent manner compared with untreated cultures , whereas no significant changes in PR isoform contents were observed in normal myometrial cells . CONCLUSIONS : These results suggest that DB05366 down-regulates P15692 , P35318 and their receptor contents and modulates PR isoform contents in cultured leiomyoma cells in a cell-type-specific manner . DB01240 -IP signaling and prostaglandin E2-EP2/EP4 signaling both mediate joint inflammation in mouse collagen-induced arthritis . Prostaglandin (PG)I2 ( prostacyclin [ P06744 ] ) and DB00917 are abundantly present in the synovial fluid of rheumatoid arthritis ( RA ) patients . Although the role of DB00917 in RA has been well studied , how much DB01240 contributes to RA is little known . To examine this issue , we backcrossed mice lacking the P43119 ( IP ) to the DBA/1J strain and subjected them to collagen-induced arthritis ( CIA ) . IP-deficient ( IP-/- ) mice exhibited significant reduction in arthritic scores compared with wild-type ( WT ) mice , despite anti-collagen antibody production and complement activation similar to WT mice . IP-/- mice also showed significant reduction in contents of proinflammatory cytokines , such as interleukin ( IL ) -6 in arthritic paws . Consistently , the addition of an IP agonist to cultured synovial fibroblasts significantly enhanced P05231 production and induced expression of other arthritis-related genes . On the other hand , loss or inhibition of each PGE receptor subtype alone did not affect elicitation of inflammation in CIA . However , a partial but significant suppression of CIA was achieved by the combined inhibition of EP2 and EP4 . Our results show significant roles of both DB01240 -IP and DB00917 -EP2/EP4 signaling in the development of CIA , and suggest that inhibition of DB00917 synthesis alone may not be sufficient for suppression of RA symptoms . 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 . 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 . P10244 prevents growth arrest associated with terminal differentiation of monocytic cells . P10244 is a transcriptional regulator of gene expression and is highly homologous to c-Myb in its N-terminal DNA binding domain . However , unlike c-myb , whose expression is restricted largely to immature hematopoietic cells , B-myb mRNA has been found to be expressed in all proliferating mammalian cell lines and is clearly regulated in a cell cycle dependent manner . That c-Myb and P10244 proteins perform different roles in proliferation and/or differentiation is suggested by the redundancy of their expression . It was previously shown that degenerated c-Myb expression can inhibit P05231 induced terminal differentiation of the leukemia cell line M1 . We found that , unlike the downregulation of c-Myb protein which is an early response of progenitor M1 cells to P05231 treatment , the downregulation of P10244 occurs late , just prior to terminal differentiation and growth arrest . It was , therefore , of interest to examine the role of the murine P10244 protein in the proliferation and differentiation of the M1 cells and to compare these effects to those of c-Myb in the same system . Clones ectopically producing P10244 , like those ectopically expressing c-Myb , proliferated in the presence of the differentiation-inducing agent and did not undergo the programmed cell death which normally follows terminal macrophage differentiation . In addition , the cell-cycle distribution of M1/ P10244 cells was comparable to untreated cells . Although M1/ P10244 and M1/c-Myb clones treated with P05231 appeared quite immature , differentiation markers were demonstrated to be maintained at near normal levels ( e.g. MyD88 , Mac-2 ) , or be partially reduced in expression ( P01024 , Fc and Mac-1 receptors ) suggesting that the cells had undergone commitment to maturation , but were unable to terminally differentiate . Human mast cells express corticotropin-releasing hormone ( P06850 ) receptors and P06850 leads to selective secretion of vascular endothelial growth factor . Mast cells are critical for allergic reactions , but also for innate or acquired immunity and inflammatory conditions that worsen by stress . P06850 ( P06850 ) , which activates the hypothalamic-pituitary-adrenal axis under stress , also has proinflammatory peripheral effects possibly through mast cells . We investigated the expression of P06850 receptors and the effects of P06850 in the human leukemic mast cell ( HMC-1 ) line and human umbilical cord blood-derived mast cells . We detected mRNA for P06850 -R1alpha , 1beta , 1c , 1e , 1f isoforms , as well as P34998 protein in both cell types . P06850 -R2alpha ( but not R2beta or R2gamma ) mRNA and protein were present only in human cord blood-derived mast cells . P06850 increased DB02527 and induced secretion of vascular endothelial growth factor ( P15692 ) without tryptase , histamine , P05231 , P10145 , or P01375 release . The effects were blocked by the P34998 antagonist antalarmin , but not the Q13324 antagonist astressin 2B . P06850 -stimulated P15692 production was mediated through activation of adenylate cyclase and increased DB02527 , as evidenced by the fact that the effect of P06850 was mimicked by the direct adenylate cyclase activator forskolin and the cell-permeable DB02527 analog 8-bromo- DB02527 , whereas it was abolished by the adenylate cyclase inhibitor SQ22536 . This is the first evidence that mast cells express functional P06850 receptors and that P06850 can induce P15692 secretion selectively . P06850 -induced mast cell-derived P15692 could , therefore , be involved in chronic inflammatory conditions associated with increased P15692 , such as arthritis or psoriasis , both of which worsen by stress . LPS induces cardiomyocyte injury through calcium-sensing receptor . DB01373 -sensing receptor ( P41180 ) belongs to the family C of G-protein coupled receptors . We have previously demonstrated that P41180 could induce apoptosis of cultured neonatal rat ventricular cardiomyocytes in simulated ischemia/reperfusion . It remains unknown whether the P41180 has function in lipopolysaccharide ( LPS ) -induced myocardial injure . The aim of this study was to investigate whether the P41180 plays a role in LPS-induced myocardial injury . Cultured neonatal rat cardiomyocytes were treated with LPS , with or without pretreatment with the P41180 -specific agonist gadolinium chloride ( GdCl3 ) or the P41180 -specific antagonist NPS2390 . Release of P01375 -α and P05231 from cardiomyocytes was observed . Levels of malonaldehyde ( MDA ) , lactate dehydrogenase ( LDH ) , and activity of superoxide dismutase ( SOD ) were measured . In addition , apoptosis of the cardiomyocytes , [Ca(2+)]i and level of P41180 expression were determined . The results showed that LPS increased cardiomyocytes apoptosis , [Ca(2+)]i , MDA , LDH , P01375 -α , P05231 release , and P41180 protein expression . Compared with LPS treatment alone , pretreatment with GdCl3 further increased apoptosis of cardiomyocytes , MDA , LDH , P01375 -α , P05231 release , [Ca(2+)]i , and the expression of the P41180 protein . Conversely , pretreatment with NPS2390 decreased apoptosis of cardiomyocytes , MDA , LDH , P01375 -α , P05231 release , [Ca(2+)]i and the expression of the P41180 protein . These results demonstrate that LPS could induce cardiomyocyte injury . Moreover , LPS-induced cardiomyocyte injury was related to P41180 -mediated cardiomyocytes apoptosis , P01375 -α , P05231 release , and increase of intracellular calcium .

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interacts_with DB00762?

multiple_choice

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 . Sequence and functional analysis of cloned guinea pig and rat serotonin P28221 receptors : common pharmacological features within the P28221 receptor subfamily . This study was undertaken to investigate the pharmacology of cloned guinea pig and rat 5-hydroxytryptamine ( serotonin ; 5-HT ) 1D receptor sites . Guinea pig , rat , and mouse P28221 receptor genes were cloned , and their amino acid sequences were compared with those of the human , dog , and rabbit . The overall amino acid sequence identity between these P28221 receptors is high and varies between 86 and 99 % . The sequence homology is slightly more divergent ( 13-27 % ) in the N-terminal extracellular region of these P28221 receptors . Guinea pig and rat P28221 receptors , stably and separately expressed in rat P13671 glial cells , are negatively coupled to cyclic AMP formation upon stimulation with agonists , as previously found for cloned human P28221 receptor sites . The cyclic AMP data show some common pharmacological features for the P28221 receptors of guinea pig , rat , and human : an almost similar rank order of potency for the investigated P28221 receptor agonists , stereoselectivity for the binding affinity and agonist potency of R(+)-8-hydroxy-2-(di-n-propylamino)tetralin , and equal P28221 receptor-mediated antagonist potency for methiothepin and the 5-HT2 receptor antagonists ritanserin and ketanserin . In conclusion , the pharmacology of the cloned P28221 receptor subtype seems , unlike the P28222 receptor subtype , conserved among various mammal species such as the human , guinea pig , and rat . [ 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 . Selective P01375 -α inhibitor-induced injection site reactions . INTRODUCTION : During the last decade , many new biological immune modulators entered the market as new therapeutic principles . P01375 -α is a pro-inflammatory cytokine known to a have a key role in the pathogenic mechanisms of various immune-mediated or inflammatory diseases . P01375 -α blockers have demonstrated efficacy in large , randomized controlled clinical trials either as monotherapy or in combination with other anti-inflammatory or disease-modifying anti-rheumatic drugs . AREAS COVERED : Although generally well tolerated and safe , potential adverse events may be associated with P01375 -α inhibitor treatment . The authors will briefly review the potential adverse drug reactions and the immunological mechanisms of injection site reactions ( ISRs ) in patients treated with etanercept and adalimumab . EXPERT OPINION : Patients treated with P01375 -α inhibitors can develop ISR around the sites of injections . ' Type IV delayed type reaction ' or ' recall ISRs ' . Eosinophilic cellulitis or ' Wells syndrome ' , ' type III ' and ' type I ' reactions are reported . Long-term studies are necessary to determine the durability of response and the real risk of ISRs with DB06674 and certolizumab pegol . Further studies are also necessary to evaluate the immunogenicity of these drugs . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . Differential binding of retinol analogs to two homologous cellular retinol-binding proteins . A comparative study of the interactions of rat cellular retinol-binding protein ( P09455 ) and cellular retinol-binding protein II ( P09455 II ) with a number of synthetic phenyl-substituted analogs of DB00162 was performed using fluorescence and nuclear magnetic resonance analysis . These studies indicate that P09455 II is more sensitive to modifications of the ring moiety than P09455 . Removal of the two methyl substituents on the ring which are ortho to the polyene chain abolishes binding to P09455 II . Conformational analysis of the ligands indicates that these two methyl groups influence the planarity of the ligand . The identification of monospecific ligands may prove useful for studying the physiological roles of these two proteins . Inhibition of the trigemino-vascular system with P28221 agonist drugs : selectively targeting additional sites of action . Inappropriate activation of the trigemino-vascular system is thought to be important in the pathogenesis of a migraine attack . The P28221 agonist sumatriptan , which is highly effective in the acute treatment of migraine , inhibits trigemino-vascular activation in animals , although its actions are normally limited to peripheral components of the trigemino-vascular system . DB00315 , a novel P28221 agonist drug , which is also highly effective in the acute treatment of migraine , acts not only at these sites , but , additionally within the brainstem , inhibiting trigemino-vascular activation centrally as well as peripherally . This article describes the pre-clinical development of DB00315 and considers , specifically , the approaches taken in the design of a molecule with attributes which facilitate access to brainstem components of the trigeminal pathway and combine this with good oral bioavailability . Superantigen-induced peripheral T-cell deletion : the effects of chemical modification of antigen-presenting cells , interleukin-4 and glucocorticoid hormones . Experiments were performed to evaluate the role of antigen-presenting cells ( P25054 ) and the effect of interleukin-4 ( P05112 ) and glucocorticoid hormone ( P30793 ) exposure on the in vitro deletion of P01730 + CD8- and CD8+ P01730 - T cells by staphylococcal enterotoxin B ( Q9Y6X0 ) . P25054 fixation with the chemical cross-linker 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide ( ECDI ) inhibited their capacity to induce Q9Y6X0 -specific deletion of mature T lymphocytes . Deletion was not influenced by treatment with anti- P10747 antibodies , which modulate T-cell activation . However , it was augmented by P05112 , known to counteract anti-CD3- and P30793 -induced thymocyte apoptosis , and was inhibited by dexamethasone ( DEX ) . These results indicate that metabolically active P25054 are required for deletion of antigen-specific mature T cells and suggest that P05112 and P30793 can modulate this phenomenon in vitro . 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 . Autophagy is induced through the ROS- P04637 - Q8N682 pathway in response to mitochondrial protein synthesis inhibition . Mitoribosome in mammalian cells is responsible for synthesis of 13 mtDNA-encoded proteins , which are integral parts of four mitochondrial respiratory chain complexes ( I , III , IV and V ) . O75616 is a nuclear-encoded GTPase important for the formation of the 28S small mitoribosomal subunit . Here , we demonstrate that knockdown of O75616 by RNA interference inhibits mitochondrial protein synthesis and promotes reactive oxygen species ( ROS ) generation , leading to autophagic vacuolization in HeLa cells . Cells that lack O75616 expression showed a significant conversion of LC3-I to LC3-II and an enhanced accumulation of autophagic vacuoles carrying the LC3 marker , all of which were blocked by the autophagy inhibitor 3-MA as well as by the ROS scavenger Q9C000 . Inhibition of mitochondrial protein synthesis either by O75616 siRNA or chloramphenicol ( CAP ) , a specific inhibitor of mitoribosomes , induced autophagy in HTC-116 P04637 ( +/+ ) cells , but not in HTC-116 P04637 ( -/- ) cells , indicating that tumor protein 53 ( P04637 ) is essential for the autophagy induction . The ROS elevation resulting from mitochondrial protein synthesis inhibition induced P04637 expression at transcriptional levels by enhancing P04637 promoter activity , and increased P04637 protein stability by suppressing P04637 ubiquitination through Q16539 /p38 MAPK-mediated P04637 phosphorylation . Upregulation of P04637 and its downstream target gene Q8N682 , but not P38936 / P38936 , was required for the autophagy induction in O75616 siRNA or CAP-treated cells . Altogether , these data indicate that autophagy is induced through the ROS- P04637 - Q8N682 pathway in response to mitochondrial protein synthesis inhibition . 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 . Genetically rescued tetrahydrobiopterin-depleted mice survive with hyperphenylalaninemia and region-specific monoaminergic abnormalities . One of the possibly mutated genes in DOPA-responsive dystonia ( DRD , Segawa 's disease ) is the gene encoding P30793 , which is the rate-limiting enzyme for tetrahydrobiopterin ( BH4 ) biosynthesis . Based on our findings on 6-pyruvoyltetrahydropterin synthase ( Q03393 ) gene-disrupted ( Pts(-/-) ) mice , we suggested that the amount of tyrosine hydroxylase ( TH ) protein in dopaminergic nerve terminals is regulated by the intracellular concentration of BH4 . In this present work , we rescued Pts(-/-) mice by transgenic introduction of human Q03393 cDNA under the control of the dopamine beta-hydroxylase promoter to examine regional differences in the sensitivity of dopaminergic neurons to BH4-insufficiency . The DPS-rescued ( Pts(-/-) , DPS ) mice showed severe hyperphenylalaninemia . Human Q03393 was efficiently expressed in noradrenergic regions but only in a small number of dopaminergic neurons . DB03886 and dopamine contents , and TH activity in the striatum were poorly restored compared with those in the midbrain . TH-immunoreactivity in the lateral region of the striatum was far weaker than that in the medial region or in the nucleus accumbens . We concluded that dopaminergic nerve terminals projecting to the lateral region of the striatum are the most sensitive to BH4-insufficiency . Biochemical and pathological changes in DPS-rescued mice were similar to those in human malignant hyperphenylalaninemia and DRD . P00747 interactions with platelets in plasma . In this report we used a fluorescent flow cytometry-based assay to examine plasminogen binding to platelets in plasma . Our data indicate that platelets activated in platelet-rich plasma ( PRP ) by adenosine-5'-diphosphate ( ADP ) or thrombin bind plasminogen to their surface . Fab fragments of the monoclonal antibody LJ-CP8 that are directed against the fibrinogen binding site on the glycoprotein ( GP ) IIb-IIIa complex inhibit both plasminogen and fibrinogen binding to ADP-stimulated platelets as does 5 mmol/L DB00974 . Platelet aggregation and plasminogen and fibrinogen binding are also concurrently inhibited by the DB00145 - DB00125 - DB00128 ( RGD ) analogue DB00145 - DB00125 - DB00145 - DB00128 - DB00133 ( GRGDS ) when it is added to PRP before ADP stimulation . The scrambled peptide analogue SDGRG has no effect . The monoclonal antibody 6D1 , directed against the P04275 binding site on GPIb , has no effect on plasminogen-platelet binding , nor does antithrombospondin antibody . epsilon- DB00513 ( DB00513 ) , however , inhibits plasminogen binding to ADP-activated platelets . These data indicate that plasminogen binds to platelets activated in plasma , that binding occurs on platelet P08514 /IIIa , and that binding may be mediated via plasminogen association with fibrinogen via lysine binding domains . Finally , we found both plasminogen and fibrinogen on resting platelets in PRP and demonstrated that they are equally displaced by DB00974 , LJ-CP8 , and 10E5 ( an additional anti- P08514 /IIIa monoclonal antibody ) . P00747 is also equally displaced by DB00513 . These data suggest that plasminogen is also bound to P08514 /IIIa on resting platelets , possibly also via interaction with fibrinogen . Enhanced sensitivity to irinotecan by Cdk1 inhibition in the p53-deficient HT29 human colon cancer cell line . Mutations in the tumor-suppressor gene p53 have been associated with advanced colorectal cancer ( CRC ) . DB00762 ( CPT-11 ) , a P11387 inhibitor , has been recently incorporated to the adjuvant therapy . Since the DNA-damage checkpoint depends on p53 activation , the status of p53 might critically influence the response to CPT-11 . We analysed the sensitivity to CPT-11 in the human colon cancer cell line HT29 ( mut p53 ) and its wild-type (wt)-p53 stably transfected subclone HT29-A4 . Cell-cycle analysis in synchronised cells demonstrated the activation of transfected wt-p53 and a P38936 ( P38936 /CIP1)-dependent cell-cycle blockage in the S phase . Activated wt-p53 increased apoptosis and enhanced sensitivity to CPT-11 . In p53-deficient cells , cDNA-macroarray analysis and western blotting showed an accumulation of the cyclin-dependent kinase (cdk)1/cyclin B complex . Subsequent p53-independent activation of the cdk-inhibitor ( cdk-I ) P38936 ( P38936 /CIP1) prevented cell-cycle progression . Cdk1 induction was exploited in vivo to improve the sensitivity to CPT-11 by additional treatment with the cdk-I P99999 -202 . We demonstrate a gain of sensitivity to CPT-11 in a p53-mutated colon cancer model either by restoring wild-type p53 function or by sequential treatment with cdk-Is . Considering that mutations in p53 are among the most common genetic alterations in CRC , a therapeutic approach specifically targeting p53-deficient tumors could greatly improve the treatment outcomes . Saccharomyces cerevisiae elongation factor 2 . Mutagenesis of the histidine precursor of diphthamide yields a functional protein that is resistant to diphtheria toxin . Protein synthesis elongation factor 2 ( P13639 ) is the target of the ADP-ribosylating activity of diphtheria toxin which is responsible for cell killing . DB03223 , an unique post-translationally modified histidine residue , is both required for and the site of this ADP-ribosylation . Although present in the P13639 of all eukaryotes and archaebacteria , the function of diphthamide is unknown . Here we describe the site-specific mutagenesis of the histidine precursor of diphthamide , histidine 699 , in yeast P13639 . Plasmid-borne EFT was randomly mutagenized at the histidine 699 codon , and the technique of plasmid shuffling was utilized to select strains that were maintained by the mutant EFT . These mutants were screened for diphtheria toxin resistance . Sequence analysis of the EFT in 49 toxin-resistant isolates showed that histidine 699 had been replaced by 1 of 4 amino acids : asparagine , glutamine , leucine , or methionine . All 11 of the possible codons corresponding to these 4 amino acids were found . The growth rates of cells sustained by the mutant forms of P13639 were slightly slower than those of isogenic wild-type cells . We conclude that despite its strict conservation and universal post-translational modification , the histidine precursor of diphthamide is not essential to the function of yeast P13639 in protein synthesis . 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 . DB05822 , a nitric oxide-releasing aspirin derivative , exhibits a significant antiproliferative effect and alters cell cycle progression in human colon adenocarcinoma cell lines . DB00435 -releasing non-steroidal antiinflammatory drugs ( NO-NSAIDs ) are safer than NSAIDs due to their ability to reduce gastric toxicity . We assessed the cytotoxic activity of a new aspirin derivative , DB05822 , after different exposure schedules , in three human colon adenocarcinoma cell lines . All the lines were positive for P23219 protein and mRNA , as evaluated by Western blot and RT-PCR , respectively , while only one was positive for P35354 . The cytostatic and cytocidal activity was determined by sulforhodamine B assay and evaluated according to Monks ' model . Cytostatic activity was observed after a 24-h drug exposure and 50 % growth inhibition was reached at concentrations ranging from 165 to 250 micro M in all cell lines , whereas with aspirin the IC50 was never reached , even at the maximum concentration tested ( 500 micro M ) , and was independent of P23219 or P35354 status . Cytocidal activity was observed only at the highest concentrations and persisted for a long time after drug removal . Flow cytometric analysis showed that the NO-aspirin compound induced a persistent accumulation of cells in G2-M phase in all the cell lines after at least 48 h exposure . Specifically , the block pertained mainly to G2 phase , whereas mitotic index was not affected at all . Our results indicate that DB05822 has an in vitro cytostatic activity superior to that of its parental aspirin compound , which makes it a potentially important tumor preventive agent . Furthermore , the cytocidal effect observed at the highest concentrations and the induction of a specific block in G2 phase renders it a promising candidate for drug combination treatments . 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 % . 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 . Chemopreventive effects of the dietary histone deacetylase inhibitor tributyrin alone or in combination with vitamin A during the promotion phase of rat hepatocarcinogenesis . The chemopreventive effects of tributyrin ( TB ) and vitamin A ( VA ) , alone or in combination , were investigated during the promotion phase of rat hepatocarcinogenesis . Compared to diethylnitrosamine control rats , TB and TB+VA-treated rats , but not VA-treated rats , presented a lower incidence and mean number of hepatocyte nodules and a smaller size of persistent preneoplastic lesions ( pPNLs ) . In addition , TB and TB+VA-treated rats exhibited a higher apoptotic body index in pPNL and remodeling PNL , whereas VA-treated rats presented only a higher apoptotic body index in remodeling PNL . None of the treatments inhibited cell proliferation in PNL . TB and TB+VA-treated rats , but not VA-treated rats , exhibited higher levels of H3K9 acetylation and P38936 protein expression . TB and VA-treated rats exhibited increased hepatic concentrations of butyric acid and retinoids , respectively . Compared to normal rats , diethylnitrosamine control animals exhibited lower retinyl palmitate hepatic concentrations . All groups had similar expression levels and exhibited similar unmethylated P09455 promoter region in microdissected pPNL , indicating that epigenetic silencing of this gene was not involved in alteration of retinol metabolism in early hepatocarcinogenesis . Data support the effectiveness of TB as a dietary histone deacetylase inhibitor during the promotion phase of hepatocarcinogenesis , which should be considered for chemoprevention combination strategies .

MH_train_1541

MH_train_1541

MH_train_1541

interacts_with DB01656?

multiple_choice

Genome-wide association studies identify P30532 /3 and Q13639 in the development of airflow obstruction . RATIONALE : Genome-wide association studies ( GWAS ) have identified loci influencing lung function , but fewer genes influencing chronic obstructive pulmonary disease ( P48444 ) are known . OBJECTIVES : Perform meta-analyses of GWAS for airflow obstruction , a key pathophysiologic characteristic of P48444 assessed by spirometry , in population-based cohorts examining all participants , ever smokers , never smokers , asthma-free participants , and more severe cases . METHODS : Fifteen cohorts were studied for discovery ( 3,368 affected ; 29,507 unaffected ) , and a population-based family study and a meta-analysis of case-control studies were used for replication and regional follow-up ( 3,837 cases ; 4,479 control subjects ) . Airflow obstruction was defined as Q99581 (1) and its ratio to FVC ( Q99581 (1)/FVC ) both less than their respective lower limits of normal as determined by published reference equations . MEASUREMENTS AND MAIN RESULTS : The discovery meta-analyses identified one region on chromosome 15q25.1 meeting genome-wide significance in ever smokers that includes A2RU49 , P48200 , and P30532 / P32297 genes . The region was also modestly associated among never smokers . Gene expression studies confirmed the presence of P30532 /3 in lung , airway smooth muscle , and bronchial epithelial cells . A single-nucleotide polymorphism in Q13639 , a gene previously related to Q99581 (1)/FVC , achieved genome-wide statistical significance in combined meta-analysis . Top single-nucleotide polymorphisms in Q9H013 , P10826 , O14495 , and Q8TE59 were nominally replicated in the P48444 meta-analysis . CONCLUSIONS : These results suggest an important role for the P30532 /3 region as a genetic risk factor for airflow obstruction that may be independent of smoking and implicate the Q13639 gene in the etiology of airflow obstruction . 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 . Molecular discrimination of responders and nonresponders to anti- P01375 alpha therapy in rheumatoid arthritis by etanercept . INTRODUCTION : About 30 % of rheumatoid arthritis patients fail to respond adequately to TNFalpha-blocking therapy . There is a medical and socioeconomic need to identify molecular markers for an early prediction of responders and nonresponders . METHODS : RNA was extracted from peripheral blood mononuclear cells of 19 rheumatoid arthritis patients before the first application of the TNFalpha blocker etanercept as well as after 72 hours . Clinical response was assessed over 3 months using the 28-joint-count Disease Activity Score and X-ray scans . Supervised learning methods were applied to Affymetrix Human Genome U133 microarray data analysis to determine highly selective discriminatory gene pairs or triplets with prognostic relevance for the clinical outcome evinced by a decline of the 28-joint-count Disease Activity Score by 1.2 . RESULTS : Early downregulation of expression levels secondary to TNFalpha neutralization was associated with good clinical responses , as shown by a decline in overall disease activity 3 months after the start of treatment . Informative gene sets include genes ( for example , P25963 , P13236 , P10145 , P01584 , P21580 , Q07343 , O75807 and P35318 ) involved in different pathways and cellular processes such as TNFalpha signalling via NFkappaB , NFkappaB-independent signalling via DB02527 , and the regulation of cellular and oxidative stress response . Pairs and triplets within these genes were found to have a high prognostic value , reflected by prediction accuracies of over 89 % for seven selected gene pairs and of 95 % for 10 specific gene triplets . CONCLUSION : Our data underline that early gene expression profiling is instrumental in identifying candidate biomarkers to predict therapeutic outcomes of anti-TNFalpha treatment regimes . Phosphorylation of human P26358 : implication of cyclin-dependent kinases . DNA methylation plays a central role in the epigenetic regulation of gene expression during development and progression of cancer diseases . The inheritance of specific DNA methylation patterns are acquired in the early embryo and are specifically maintained after cellular replication via the DNA methyltransferase 1 ( P26358 ) . Recent studies have suggested that the enzymatic activity of P26358 is possibly modulated by phosphorylation of serine/threonine residues located in the N-terminal domain of the enzyme . In the present work , we report that cyclin-dependent kinases ( CDKs ) 1 , 2 and 5 can phosphorylate Ser154 of human P26358 in vitro . Further evidence of phosphorylation of endogenous P26358 at position 154 by CDKs is also found in 293 cells treated with roscovitine , a specific inhibitor of P06493 , 2 and 5 . To determine the importance of Ser154 phosphorylation , a mutant of P26358 encoding a single-point mutation at position 154 ( S154A ) was generated . This mutation induced a severe loss of enzymatic activity when compared to wild type P26358 . Moreover , after treatment with 5-Aza- DB02594 ( 5-aza-dC ) , a faster decline in P26358 protein level was observed for P29320 -293 cells expressing P26358 (S154A) as compared to cells expressing wild type P26358 . Our data suggest that phosphorylation of P26358 at Ser154 by CDKs is important for enzymatic activity and protein stability of P26358 . Considering that tumour-associated cell cycle defects are often mediated by alterations in CDK activity , our results suggest that dysregulation of cell cycle via CDKs could induce abnormal phosphorylation of P26358 and lead to DNA hypermethylation often observed in cancer cells . Steady-state kinetic and inhibition studies of the mammalian target of rapamycin ( P42345 ) kinase domain and P42345 complexes . The mammalian target of rapamycin ( P42345 ) is a DB00133 / DB00156 protein kinase and a major controller of cell growth . In cells , P42345 forms two distinct multiprotein complexes , mTORC1 and mTORC2 . The mTORC1 complex can phosphorylate 4EBP1 and P23443 , two key regulators of translation initiation , whereas mTORC2 phosphorylates P31749 , an event required for P31749 activation . Here , we expressed and purified human mTORC1 and mTORC2 from P29320 -293 cells using FLAG-M2 affinity chromatography . Western blotting analysis using phospho-specific antibodies indicated that recombinant mTORC1 and mTORC2 exhibit distinct substrate preferences in vitro , consistent with their roles in cells . To improve our understanding of the enzymatic properties of P42345 alone and P42345 in its complex form , steady-state kinetic profiles of truncated P42345 containing the kinase domain ( residues 1360-2549 ) and mTORC1 were determined . The results revealed that mTORC1 is catalytically less active than truncated P42345 , as evidenced by 4.7- and 3.1-fold decreases in catalytic efficiency , k(cat)/K(m) , for DB00171 and 4EBP1 , respectively . We also found that truncated P42345 undergoes autophosphorylation through an intramolecular mechanism . Mass spectrometric analysis identified two novel P42345 autophosphorylation sites , Ser2454 and either Thr2473 or Thr2474 , in addition to the previously reported Ser2481 site . Truncated P42345 and mTORC1 were completely inhibited by DB00171 competitive inhibitors PI103 and BEZ235 and partially inhibited by rapamycin/ P62942 in a noncompetitive fashion toward DB00171 . All inhibitors tested exhibited similar inhibitory potencies between mTORC1 and truncated P42345 containing the kinase domain . Our studies presented here provide the first detailed kinetic studies of a recombinant P42345 complex . Levels of angiopoietins 1 and 2 in induced sputum supernatant in patients with P48444 . Pathological features of chronic obstructive pulmonary disease ( P48444 ) include lung vascular remodeling and angiogenesis . Q15389 ( Ang-1 ) , is an essential mediator of angiogenesis by establishing vascular integrity , whereas angiopoietin-2 ( Ang-2 ) acts as its natural inhibitor . We determined the levels of angiopoietins in sputum supernatants of patients with P48444 and investigated their possible association with mediators and cells involved in the inflammatory and remodeling process . Fifty-nine patients with P48444 , 25 healthy smokers and 20 healthy non-smokers were studied . All subjects underwent lung function tests , sputum induction for cell count identification and Ang-1 , Ang-2 , P15692 , TGF-β1 , P08253 , LTB4 , P10145 , albumin measurement in sputum supernatants . Airway vascular permeability ( AVP ) index was also assessed . Ang-2 levels were significantly higher in patients with P48444 compared to healthy smokers and healthy non-smokers [ median , interquartile ranges pg/ml , 267 ( 147-367 ) vs. 112 ( 67-171 ) and 98 ( 95-107 ) , respectively ; p < 0.001 ] . Regression analysis showed a significant association between Ang-2 levels and AVP index , P15692 , P10145 and P08253 levels in P48444 , the strongest being with P15692 . Our results indicate that induced sputum Ang-2 levels are higher in P48444 compared to healthy smokers and healthy non-smokers . Moreover , Ang-2 is associated with AVP , P10145 , P08253 , and P15692 , indicating a possible role for Ang-2 in the pathogenesis of the disease . 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 . 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 . Specific Biomarkers Are Associated with Docetaxeland Gemcitabine-Resistant NSCLC Cell Lines . Five-year survival rate for lung cancer is limited to 10 % to 15 % . Therefore , the identification of novel therapeutic prognostic factors is an urgent requirement . The aim of this study is thus to highlight specific biomarkers in chemoresistant non-small cell lung cancer cell lines . Therefore , we checked-in the control condition as well as after short-term pharmacological treatment with either docetaxel or gemcitabine-the expression of genes such as tumor suppressor genes ( CDKN2A , P53355 , P49789 , P09211 , P16455 , RARβ2 , RASSF1A , and P35625 ) , genes associated with drug resistance ( P38398 , P35354 , P07992 , P17936 , P23921 , and Q13509 ) , and stemness-related genes ( CD133 , Q01860 , and O43623 ) in two cellular models of squamous carcinoma ( CAEP ) and adenocarcinoma ( RAL ) of the lung originally established . Their promoter methylation profile was also evaluated . Drug-related genes were upregulated . DB00515 resistance matched with high levels of P38398 and P07992 in both cell lines ; docetaxel sensitivity of CAEP cells was associated to levels of Q13509 lower than RAL cells . Although CAEP cells were more sensitive to gemcitabine , both cell lines showed high levels of P23921 . Stemness-related genes were downregulated in the control condition but became upregulated in docetaxel-resistant cells , indicating the selection of a population with stemness features . We did not find an unequivocal correspondence between gene expression and respective DNA promoter methylation status , suggesting the involvement of additional mechanisms of gene expression regulation . These results highlight specific biomarkers consistent with the different responses of the two cell lines to standard pharmacological treatments and indicate specific molecular traits for their chemoresistance . Glycine-N methyltransferase expression in HepG2 cells is involved in methyl group homeostasis by regulating transmethylation kinetics and DNA methylation . Glycine-N methyltransferase ( Q14749 ) is a potential tumor suppressor that is commonly inactivated in human hepatoma . We systematically investigated how Q14749 regulates methyl group kinetics and global DNA methylation . HepG2 cells ( Q14749 inactive , Q14749 - ) and cells transfected with Q14749 expressed vector ( Q14749 + ) were cultured in low ( 10 μmol/L ) , adequate ( 100 μmol/L ) , or high ( 500 μmol/L ) l-methionine , each with 2.27 μmol/L folate . Transmethylation kinetics were studied using stable isotopic tracers and GC-MS . Methylation status was determined by S-adenosylmethionine ( DB00118 ) and S-adenosylhomocysteine ( Q53FZ2 ) levels , DB00118 : Q53FZ2 ratio , DNA methyltransferase ( P26358 ) activity , and methylated cytidine levels in DNA . Compared with Q14749 - cells , Q14749 + cells had lower homocysteine and greater cysteine concentrations . Q14749 expression increased methionine clearance by inducing homocysteine transsulfuration and remethylation metabolic fluxes when cells were cultured in high or adequate l-methionine . In contrast , homocysteine remethylation flux was lower in Q14749 + cells than in Q14749 - cells and homocysteine transsulfuration fluxes did not differ when cells were cultured in low methionine , suggesting that normal Q14749 function helps to conserve methyl groups . Furthermore , Q14749 expression decreased DB00118 and increased Q53FZ2 levels and reduced P26358 activity in high or adequate , but not low , methionine cultures . In low methionine cultures , restoring Q14749 in HepG2 cells did not lead to sarcosine synthesis , which would waste methyl groups . Methylated cytidine levels were significantly lower in Q14749 - cells than in Q14749 + cells . In conclusion , we have shown that Q14749 affects transmethylation kinetics and DB00118 synthesis and facilitates the conservation of methyl groups by limiting homocysteine remethylation fluxes . DB00864 -induced hypertension : what 's endothelial and hematopoietic P62942 got to do with it ? Biomarker analysis of neoadjuvant doxorubicin/cyclophosphamide followed by ixabepilone or Paclitaxel in early-stage breast cancer . PURPOSE : Predictive biomarkers offer the potential to improve the benefit:risk ratio of a therapeutic agent . DB04845 achieves comparable pathologic complete response ( pCR ) rates to other active drugs in the neoadjuvant setting . This phase II trial was designed to investigate potential biomarkers that differentiate response to this agent . EXPERIMENTAL DESIGN : Women with untreated , histologically confirmed primary invasive breast adenocarcinoma received neoadjuvant doxorubicin/cyclophosphamide , followed by 1:1 randomization to ixabepilone ( n = 148 ) or paclitaxel ( n = 147 ) . Rates of pCR were compared between treatment arms based on predefined biomarker sets : Q13509 , Q9Y6A5 , and CAPG gene expression , a 20- and 26-gene expression model , P08183 protein expression , and other potential markers of sensitivity . βIII-tubulin protein expression is reported separately but is referred to here for completeness . All patients underwent a core needle biopsy of the primary cancer for molecular marker analysis before chemotherapy . Gene expression profiling data were used for molecular subtyping . RESULTS : There was no significant difference in the rate of pCR in both treatment arms in βIII-tubulin-positive patients . Higher pCR rates were observed among βIII-tubulin-positive patients than in βIII-tubulin-negative patients . Furthermore , no correlation was evident between Q13509 , Q9Y6A5 , and CAPG gene expression , P08183 protein expression , multi-gene expression models , and the efficacy of ixabepilone or paclitaxel , even within the estrogen receptor-negative subset . CONCLUSION : These results indicate that βIII-tubulin protein and mRNA expression , P08183 protein expression , Q9Y6A5 and CAPG gene expression , and multigene expression models ( 20- and 26-gene ) are not predictive markers for differentiating treatment benefit between ixabepilone and paclitaxel in early-stage breast cancer . 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 . The combination of axitinib followed by paclitaxel/carboplatin yields extended survival in advanced P15056 wild-type melanoma : results of a clinical/correlative prospective phase II clinical trial . BACKGROUND : Simultaneous chemotherapy with vascular endothelial growth factor ( P15692 ) inhibition has not shown additional benefit over chemotherapy alone in advanced melanoma . We tested administration of the potent P15692 inhibitor axitinib followed by paclitaxel/carboplatin to determine whether enhanced tumour proliferation during axitinib withdrawal leads to sustained chemosensitivity . METHODS : We conducted a prospective phase II trial in metastatic melanoma patients with ECOG performance status 0-1 and normal organ function . DB06626 5 mg PO b.i.d. was taken on days 1-14 of each 21-day treatment cycle , and carboplatin ( AUC=5 ) with paclitaxel ( 175 mg m(-2) ) was administered on day 1 starting with cycle 2 . 3'-Deoxy-3'-(18)F-fluorothymidine ( (18)F- P17948 ) -PET scans were performed in five patients to assess tumour proliferation on days 1 , 14 , 17 , and 20 of cycle 1 . Molecular profiling for P15056 was performed for all patients with cutaneous , acral , or mucosal melanoma . RESULTS : The treatment was well tolerated . The most common grade 3 AEs were hypertension , neutropenia , and anaemia . Grade 4 non-haematologic AEs were not observed . Four of five patients completing (18)F- P17948 -PET scans showed increases ( 23-92 % ) in SUV values during the axitinib holiday . Of 36 evaluable patients , there were 8 confirmed PRs by Response Evaluation Criteria in Solid Tumors . Overall , 20 patients had SD and 8 had PD as the best response . The median PFS was 8.7 months and the median overall survival was 14.0 months . Five P15056 (V600E/K) patients had significantly worse PFS than patients without these mutations . CONCLUSIONS : DB06626 followed by carboplatin and paclitaxel was well tolerated and effective in P15056 wild-type metastatic melanoma . 3'-Deoxy-3'-(18)F-fluorothymidine-PET scans showed increased proliferation during axitinib withdrawal . Solution structure and backbone dynamics of the catalytic domain of matrix metalloproteinase-2 complexed with a hydroxamic acid inhibitor . P08253 is a member of the matrix metalloproteinase family that has been implicated in tumor cell metastasis and angiogenesis . Here , we describe the solution structure of a catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) , determined by three-dimensional heteronuclear NMR spectroscopy . The catalytic domain , designated MMP-2C , has a short peptide linker replacing the internal fibronectin-domain insertion and is enzymatically active . Distance geometry-simulated annealing calculations yielded 14 converged structures with atomic root-mean-square deviations ( r.m.s.d. ) of 1.02 and 1.62 A from the mean coordinate positions for the backbone and for all heavy atoms , respectively , when 11 residues at the N-terminus are excluded . The structure has the same global fold as observed for other MMP catalytic domains and is similar to previously solved crystal structures of P08253 . Differences observed between the solution and the crystal structures , near the bottom of the S1 ' specificity loop , appear to be induced by the large inhibitor present in the solution structure . The MMP-2C solution structure is compared with P22894 crystal structure bound to the same inhibitor to highlight the differences especially in the S1 ' specificity loop . The finding provides a structural explanation for the selectivity between P08253 and P22894 that is achieved by large inhibitors . Receptor selectivity of the cloned opossum G protein-coupled receptor kinase 2 ( P25098 ) in intact opossum kidney cells : role in desensitization of endogenous alpha2C-adrenergic but not serotonin 1B receptors . To characterize the specificity of endogenously expressed G protein-coupled receptor kinases ( GRKs ) for endogenous Gi-coupled alpha2C-adrenergic and serotonin 1B ( P28222 ) receptors in the opossum kidney ( OK ) cell line , we have isolated a 3.073-kb OK- P25098 clone encoding a 689-amino acid protein that shares 94.2 % amino acid identity with rat P25098 . Northern blot analysis revealed the presence of P25098 mRNA transcripts of 5.0 and 3.0 kb in OK cells . In intact OK cells , preincubation ( 45 min ) with agonist ( 5-HT or UK 14304 , 1 microM ) reduced the maximal inhibition of forskolin-induced DB02527 accumulation mediated by endogenous P28222 and alpha2C-adrenergic receptors by 12 +/- 2 % or 17 +/- 4 % , respectively . In transfected OK cells overexpressing OK- P25098 , agonist-induced desensitization of the alpha2C-adrenergic receptor , but not the P28222 receptor , was enhanced by 2- to 4-fold . Conversely , in cells overexpressing the kinase-inactive mutant OK- P25098 -K220R , alpha2C-adrenergic receptor desensitization was selectively abolished , whereas desensitization of the P28222 receptor was slightly enhanced . Similarly , depletion of GRK-2 protein by stable transfection of full-length antisense OK- P25098 cDNA blocked the desensitization of alpha2C-adrenergic receptors but not of P28222 receptors . These results represent the first evidence of the coexistence of P25098 -dependent ( for alpha2C receptors ) and P25098 -independent ( for P28222 receptors ) mechanisms of desensitization in intact cells and demonstrate the selectivity of P25098 for distinct Gi-coupled receptors . The efficacy and tolerability of frovatriptan and dexketoprofen for the treatment of acute migraine attacks . DB00998 is a DB00669 characterized by a high affinity for P28222 /1D receptors and a long half-life contributing to a more sustained and prolonged action than other triptans . DB09214 is a nonsteroidal anti-inflammatory drug with a relatively short half-life and rapid onset of action , blocking the action of cyclo-oxygenase , which is involved in prostaglandins ' production , thus reducing inflammation and pain . Both drugs have been successfully employed as monotherapies for the treatment of acute migraine attacks . The combination of these two drugs ( frovatriptan 2.5 mg plus dexketoprofen 25 or 37.5 mg ) has been tested in migraine sufferers , showing a rapid and good initial efficacy , with 2-h pain free rates of 51 % , and a high persistence in the 48-h following the onset of pain : recurrence occurred in only 29 % of attacks and sustained pain free rates were 43 % at 24- and 33 % at 48-h . P10997 -driven metabolic reprogramming induces regression of p53-deficient tumours in vivo . P04637 is commonly altered in human cancer , and Tp53 reactivation suppresses tumours in vivo in mice ( P04637 and Tp53 are also known as p53 ) . This strategy has proven difficult to implement therapeutically , and here we examine an alternative strategy by manipulating the p53 family members , Tp63 and Tp73 ( also known as p63 and p73 , respectively ) . The acidic transactivation-domain-bearing ( TA ) isoforms of p63 and p73 structurally and functionally resemble p53 , whereas the ΔN isoforms ( lacking the acidic transactivation domain ) of p63 and p73 are frequently overexpressed in cancer and act primarily in a dominant-negative fashion against p53 , TAp63 and TAp73 to inhibit their tumour-suppressive functions . The p53 family interacts extensively in cellular processes that promote tumour suppression , such as apoptosis and autophagy , thus a clear understanding of this interplay in cancer is needed to treat tumours with alterations in the p53 pathway . Here we show that deletion of the ΔN isoforms of p63 or p73 leads to metabolic reprogramming and regression of p53-deficient tumours through upregulation of P10997 , the gene that encodes amylin , a 37-amino-acid peptide co-secreted with insulin by the β cells of the pancreas . We found that P10997 is causally involved in this tumour regression and that amylin functions through the calcitonin receptor ( CalcR ) and receptor activity modifying protein 3 ( O60896 ) to inhibit glycolysis and induce reactive oxygen species and apoptosis . DB01278 , a synthetic analogue of amylin that is currently used to treat type 1 and type 2 diabetes , caused rapid tumour regression in p53-deficient thymic lymphomas , representing a novel strategy to target p53-deficient cancers . [ Evaluation of occurrence frequency of circulating p53 protein in serum of patients with chronic obstructive pulmonary diseases and non-small cell lung cancer ] . THE AIM OF STUDY was the evaluation of occurrence frequency of increased concentration of p53 protein in serum of patients with P48444 or NSCLC . MATERIAL AND METHODS : Participants have been enrolled to the one of three studied groups : patients with P48444 , patients with NSCLC and healthy subjects . In each patient serum concentration of p53 protein was measured with ELISA method ( photometric immunoassay ELISA , ROCHE Molecular Biochemicals , Manheim , Germany ) . Comparative analysis of frequency of p53 occurrence in serum in three studied group has been done with respect to nicotine addiction in P48444 and NSCLC groups . Chi-squared test was used for statistical analysis . 95 % confidence interval was set for statistically significant differences . RESULTS : 164 participants was enrolled to the study including : 53 with NSCLC , 59 with P48444 and 52 healthy control . Presence of p53 protein in serum was observed in 45 % of patients with NSCLC , 34 % of patients with P48444 , and in 11.5 % healthy people . In healthy subjects increased serum concentration of p53 was found out significantly more seldom than in P48444 ( p = 0.006 ) and NSCLC ( p = 0.0001 ) groups . There was no significant difference between prevalence of p53 protein in serum of patients with NSCLC or P48444 ( p = 0.2 ) . CONCLUSIONS : P04637 protein is observed in serum three times more often in patients with P48444 than healthy subjects . Preasumably , chronic inflammation in bronchial tree in course of P48444 could be predisposing factor for p53 mutation and synthesis of pathological p53 protein . Differential actions of vasopeptidase inhibition versus angiotensin-converting enzyme inhibition on diuretic therapy in experimental congestive heart failure . BACKGROUND : DB00886 ( OMA ) , a vasopeptidase inhibitor , simultaneously inhibits angiotensin-converting enzyme ( P12821 ) and neutral endopeptidase , which degrades vasodilatory factors ( eg , P35318 ) and natriuretic peptides . Based on the beneficial cardiorenal and humoral properties of the natriuretic peptides , we hypothesized that an acute vasopeptidase inhibitor with or without diuretic would result in more favorable cardiorenal and hormonal actions than P12821 inhibition plus diuretic ( ACEI+D ) in congestive heart failure . METHODS AND RESULTS : We compared the actions of OMA alone and with diuretic ( OMA+D ) to ACEI+D in a model of pacing-induced congestive heart failure . OMA+D decreased pulmonary arterial and pulmonary capillary wedge pressures to a greater level than OMA alone or ACEI+D . Glomerular filtration rate was lower with ACEI+D than with either OMA group . Plasma renin activity and aldosterone immediately increased with ACEI+D , whereas OMA+D resulted in higher plasma renin activity and a delayed increase in aldosterone . OMA alone did not increase plasma renin activity and aldosterone , but resulted in a sustained increase in plasma adrenomedullin , with higher urinary atrial natriuretic peptide , adrenomedullin , and cGMP excretions than with ACEI+D . CONCLUSIONS : Acute administration of OMA with or without diuretic results in more favorable cardiorenal and humoral responses in experimental congestive heart failure than does ACEI+D . There is no acute activation of renin and aldosterone with OMA alone such as occurs with ACEI+D and OMA+D . Thus , OMA with or without a diuretic possesses beneficial cardiorenal and humoral actions comparable to those observed with ACEI+D that can be explained by potentiation of natriuretic peptides .

MH_train_1542

MH_train_1542

MH_train_1542

interacts_with DB01120?

multiple_choice

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 . 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 . The expression profile of DB00171 -binding cassette transporter genes in breast carcinoma . AIM : DB00171 -binding cassette ( DB01048 ) transporters contribute to development of resistance to anticancer drugs via DB00171 -dependent drug efflux . A major goal of our study was to investigate associations between the expression of ABC transporters and outcome of breast carcinoma patients . PATIENTS & METHODS : Transcript levels of all 49 human ABC transporters were determined in post-treatment tumor and non-neoplastic tissue samples from 68 breast carcinoma patients treated by neoadjuvant chemotherapy . Six ABC transporters were then evaluated in independent series of 100 pretreatment patients . RESULTS : Q8WWZ7 /6/8/9/10 , P08183 /5/11 , O95255 /9 , Q9UBJ2 /4 , Q9H222 and Q9H221 were significantly downregulated and Q9BZC7 /3/7/12 , Q03518 /3/8/9/10 , P33527 /4/5/10/11/12 , P33897 /3 , P61221 , Q8NE71 /2/3 and P45844 were upregulated in post-treatment tumors compared with non-neoplastic tissues . Significant associations of intratumoral levels of P33527 and Q09428 with grade and expression of hormonal receptors were found in both sets of patients . Q86UK0 , Q86UQ4 and Q9UBJ2 levels were significantly associated with the response to neoadjuvant chemotherapy in post-treatment patients . Protein expression of Q86UK0 , Q09428 and Q9UBJ2 in tumor tissues of patients with breast carcinoma was observed by immunoblotting for the first time . CONCLUSION : Q86UK0 , Q86UQ4 , P33527 , Q09428 and Q9UBJ2 present potential modifiers of progression and response to the chemotherapy of breast carcinoma . Growth-inhibitory effects of vitamin D analogues and retinoids on human pancreatic cancer cells . Retinoids and vitamin D are important factors that regulate cellular growth and differentiation . An additive growth-inhibitory effect of retinoids and vitamin D analogues has been demonstrated for human myeloma , leukaemic and breast cancer cells . We set out to study the effects of the vitamin D analogue EB1089 and the retinoids all-trans- and 9-cis-retinoic acid on the human pancreatic adenocarcinoma cell lines Capan 1 and Capan 2 and the undifferentiated pancreatic carcinoma cell line Hs766T . The cell lines investigated expressed vitamin D receptor , retinoic acid receptor ( RAR ) -alpha and gamma as determined by polymerase chain reaction after reverse transcription . P10826 was expressed only in Hs766T cells . Addition of all-trans-retinoic acid increased the amount of P10276 mRNA in the three cell lines and induced P10826 mRNA in Capan 1 and Capan 2 cells . All-trans-retinoic acid at a concentration of 10 nM inhibited the growth of Capan 1 and Capan 2 cells by 40 % relative to controls . DB00523 was less effective . Neither all-trans-retinoic acid nor 9-cis-retinoic acid affected the growth of Hs766T cells . EB1089 , if added alone to the cells , did not significantly inhibit growth . However , the combination of 1 nM EB1089 with 10 nM all-trans-retinoic acid exerted a growth-inhibitory effect of 90 % in Capan 1 cells and of 70 % in Capan 2 cells . Our data suggest that vitamin D analogues together with retinoids inhibit the growth of human pancreatic cancer cells . However , in vivo studies are necessary to examine the potential use of retinoids and vitamin D analogues on pancreatic cancer . P15692 inhibits tumor cell invasion and mesenchymal transition through a MET/ P35968 complex . Inhibition of P15692 signaling leads to a proinvasive phenotype in mouse models of glioblastoma multiforme ( GBM ) and in a subset of GBM patients treated with bevacizumab . Here , we demonstrate that vascular endothelial growth factor ( P15692 ) directly and negatively regulates tumor cell invasion through enhanced recruitment of the protein tyrosine phosphatase 1B ( P18031 ) to a MET/ P35968 heterocomplex , thereby suppressing P14210 -dependent MET phosphorylation and tumor cell migration . Consequently , P15692 blockade restores and increases MET activity in GBM cells in a hypoxia-independent manner , while inducing a program reminiscent of epithelial-to-mesenchymal transition highlighted by a P55290 to P19022 switch and enhanced mesenchymal features . Inhibition of MET in GBM mouse models blocks mesenchymal transition and invasion provoked by P15692 ablation , resulting in substantial survival benefit . Stage-dependent expression of PI3K/Akt‑pathway genes in neuroblastoma . The phosphoinositide-3 kinase ( PI3K ) pathway plays a critical role in cancer cell growth and survival and has also been implicated in the development of the childhood cancer neuroblastoma . In neuroblastoma high mRNA expression of the PI3K catalytic isoform O00329 is associated to favorable disease . Yet , activation of Akt is associated with poor prognosis . Since the contribution of the numerous members of this pathway to neuroblastoma pathogenesis is mainly unknown , genes of the PI3K/Akt pathway were analyzed at the mRNA level through microarrays and quantitative real-time RT-PCR ( TaqMan ) and at the protein level using western blot analysis . Five genes showed lower mRNA expression in aggressive compared to more favorable neuroblastomas ( Q05513 , P05771 , Q13541 , PIK3RI and O00329 ) while the opposite was seen for P16234 . Clustering analysis shows that the expression levels of these six genes can predict aggressive disease . At the protein level , p110δ ( encoded by O00329 ) and p85α isomers ( encoded by P27986 ) were more highly expressed in favorable compared to aggressive neuroblastoma . Evaluation of the expression of these PI3K genes can predict aggressive disease , and indicates stage-dependent involvement of PI3K-pathway members in neuroblastoma . 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 . P15121 inhibitor fidarestat prevents retinal oxidative stress and vascular endothelial growth factor overexpression in streptozotocin-diabetic rats . The study addressed the role for aldose reductase ( AR ) in 1 ) retinal oxidative stress and vascular endothelial growth factor ( P15692 ) overexpression in early diabetes , and 2 ) high glucose-induced oxidative stress in retinal endothelial cells . In vivo experiments were performed on control rats and diabetic rats treated with or without low or high dose of the AR inhibitor ( Q9Y4X5 ) fidarestat ( 2 or 16 mg. kg(-1). day(-1) ) . In vitro studies were performed on bovine retinal endothelial cells ( BREC ) cultured in either 5 or 30 mmol/l glucose with or without 1 micro mol/l fidarestat . Intracellular reactive oxygen species were assessed using the 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate ( H(2)DCFDA ) probe and flow cytometry . Both low and high doses of fidarestat ( i.e. , the doses that partially and completely inhibited sorbitol pathway hyperactivity ) arrested diabetes-induced retinal lipid peroxidation . This was achieved due to upregulation of the key antioxidative defense enzyme activities rather than changes in reduced glutathione , oxidized glutathione , ascorbate and dehydroascorbate concentrations , and the glutathione and ascorbate redox states . Diabetes-associated 2.1-fold P15692 protein overexpression ( enzyme-linked immunosorbent assay ; ELISA ) was dose-dependently prevented by fidarestat , whereas total P15692 mRNA and P15692 -164 mRNA ( RT-PCR ) abundance were not affected by either diabetes or the Q9Y4X5 . In BREC , fidarestat corrected hyperglycemia-induced increase in H(2)DCFDA fluorescence but not oxidative stress caused by three different pro-oxidants in normoglycemic conditions . In conclusion , increased AR activity contributes to retinal oxidative stress and P15692 protein overexpression in early diabetes . The findings justify the rationale for evaluation of fidarestat on diabetic retinopathy . Synthesis of oleanolic acid derivatives : In vitro , in vivo and in silico studies for P18031 inhibition . Non-insulin dependent diabetes mellitus is a multifactorial disease that links different metabolic routes ; a point of convergence is the enzyme P18031 which turns off insulin and leptin receptors involved in glucose and lipid metabolism , respectively . Pentacyclic acid triterpenes such as oleanolic acid ( OA ) have proved to be excellent P18031 inhibitors , thus , the purpose of current work was to generate a series of derivatives that improve the pharmacological effect of OA . Our findings suggest that the presence of the carboxylic acid and/or its corresponding reduction product carbinol derivative ( H-bond donor ) in C-28 is required to maintain the inhibitory activity ; moreover , this is further enhanced by ester or ether formation on C-3 . The most active derivatives were cinnamoyl ester ( 6 ) and ethyl ether ( 10 ) . DB04088 showed potent in vitro inhibitory activity and significantly decrease of blood glucose levels on in vivo experiments . Meanwhile , 10 showed contrasting outcomes , since it was the compound with higher inhibitory activity and selectivity over P18031 and has improved interaction with site B , according with docking studies , the in vivo antidiabetic effect was similar to oleanolic acid . In conclusion , oleanolic acid derivatives have revealed an enhanced inhibitory effect over P18031 activity by increasing molecular interactions with either catalytic or allosteric sites and producing a hypoglycaemic effect on non insulin dependent diabetes mellitus rat model . 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 . 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 . 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 . A novel epitope from P20273 regulates Th1 and Th17 cell function in systemic lupus erythematosus . The published antibodies ( Abs ) against P20273 on B cells including DB04958 could inhibit B cell activation mainly through binding to P06681 -set Ig domain of P20273 , but they are rarely reported to modulate the pathogenic P01730 (+) T cell function in systemic lupus erythematosus ( SLE ) . Recently , it was proved that the extracellular amino-terminal V-set Ig domain of P20273 might mediate the interaction of B and T cells , but for now the exact effect of this domain on P01730 (+) T cell biology have not been identified . Thus , in this study , we screened out a peptide termed B2285 from this V-set Ig domain , developed the novel specific anti-B2285 Abs in rabbits , and investigated their effects in MRL/lpr mice with spontaneous SLE . The results showed that anti-B2285 Abs could ameliorate the disease severity obviously in spontaneous SLE mice with the decreased differentiations of Th1 and Th17 cells and no changes of Th2 and Treg cells . In co-cultured B cells and P01730 (+) T cells , this specific anti- P20273 Abs was observed to inhibit the anti-dsDNA Abs production , P01730 (+) T cells proliferation , the protein levels of T-bet and RORγt , and the mRNA levels of P01375 -α , IFN-γ , P05231 and Q16552 in P01730 (+) T cells . Moreover , the expression of CD45RO on P01730 (+) T cells could be also apparently diminished by this novel Abs . The data suggested that anti-B2285 Abs could slow SLE progression significantly by regulating Th1 and Th17 cells function via B-T cell interaction and the cytokine network regulation . The treatment against V-set Ig domain of P20273 would be a valuable therapeutic method for SLE and other autoimmune diseases . Current therapy for patients with sitosterolemia -- effect of ezetimibe on plant sterol metabolism . Sitosterolemia is a rare , autosomal recessive inherited sterol storage disease associated with high tissue and serum plant sterol concentrations , caused by mutations in the adenosine triphosphate-bind-ing cassette ( DB01048 ) transporter Q9H222 or Q9H221 genes . Markedly increased serum concentration of plant sterols. such as sitosterol and campesterol , cause premature atherosclerosis and massive xanthomas . Hitherto known treatments for sitosterolemia , including a low-sterol diet , bile-salt binding resins , ileal bypass surgery and low density lipoprotein ( LDL ) apheresis have not yielded sufficient reduction of serum plant sterol levels and many patients show a sustained elevation of plant sterol levels , subsequently developing premature atherosclerotic cardiovascular diseases . DB00973 , an inhibitor of intestinal cholesterol absorption through its binding to Niemann-Pick C1-like 1 ( Q9UHC9 ) , has been widely used for decreasing serum LDL-cholesterol levels in patients with hypercholesterolemia . DB00973 also reduces the gastrointestinal absorption of plant sterols , thereby also lowering the serum concentrations of plant sterols . This pharmacological property of ezetimibe shows its potential as a novel effective therapy for sitosterolemia . In the current review , we discuss the current therapy for patients with sitosterolemia and present two Japanese adolescent patients with this disease , one of whom underwent percutaneous coronary intervention for accelerated coronary atherosclerosis . DB00973 administration in addition to conventional drug therapy successfully reduced serum sitosterol levels by 51.3 % and 48.9 % , respectively , in the two patients , demonstrating ezetimibe as a novel and potent treatment agent for sitosterolemia that could work additively with conventional drug therapy . 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 . 4-[3,5-Bis(trimethylsilyl)benzamido] benzoic acid inhibits angiogenesis in colon cancer through reduced expression of vascular endothelial growth factor . 4-[3,5-bis(trimethylsilyl)benzamido] Benzoic acid ( TAC-101 ) has potent antiproliferative , antiangiogenic , and antitumor effects in vitro and in vivo . These effects might be due to TAC-101 binding to retinoic acid receptor alpha ( P10276 ) and interfering with the binding of activator protein-1 ( AP-1 ) to DNA . However , little is known about the detailed mechanism of TAC-101 function . We investigated the mechanism of the antiangiogenic effect of TAC-101 using a rat hepatic metastatic model in vivo and DLD-1 human colon cancer cells in vitro . Liver metastases were induced by portal injection of Q15293 -9 rat colonic cancer cells into F344 rats . TAC-101 ( 8 mg/kg ) was orally administered 5 days per week for 4 weeks and then hepatic tumors were immunohistochemically evaluated for microvessel density ( P53602 ) and vascular endothelial growth factor ( P15692 ) . TAC-101 significantly reduced both P53602 and P15692 expression . Northern blot analysis and ELISA indicated that TAC-101 efficiently inhibited production of P15692 mRNA and protein in DLD-1 cells in a time- and dose-dependent manner . These findings suggest that TAC-101 may inhibit progression and metastasis in colon cancer by interfering with tumor production of P15692 . The antiproliferative effect of lidocaine on human tongue cancer cells with inhibition of the activity of epidermal growth factor receptor . Local anesthetics suppress proliferation in several cancer cells . The mechanism of the suppression , however , is unknown . Our previous study shows that lidocaine , at the level of tissue concentration under topical or local administration , has a direct inhibitory effect on the activity of epidermal growth factor receptor ( P00533 ) , which is a potential target for antiproliferation in cancer cells . Therefore , we hypothesized that lidocaine would suppress the proliferation of cancer cells through the inhibition of P00533 activity . We investigated the effects of lidocaine ( 40-4000 microM ) on proliferation of a human tongue cancer cell line , CAL27 , which has a high level of P00533 expression , and also examined the effect of lidocaine on epidermal growth factor ( P01133 ) -stimulated autophosphorylation of P00533 in CAL27 cells . A clinical concentration of lidocaine ( 400 microM ) suppressed both serum-induced and P01133 -induced proliferation of CAL27 cells and inhibited P01133 -stimulated tyrosine kinase activity of P00533 without cytotoxicity . A larger concentration of lidocaine ( 4000 microM ) showed cytotoxicity with an antiproliferative effect . We suggest that the inhibition of P01133 -stimulated P00533 activity is one of the mechanisms of the antiproliferative effect of lidocaine on CAL27 cells . DB00281 administered topically within the oral cavity for cancer pain relief may suppress the proliferation of human tongue cancer cells . The thioredoxin reductase inhibitor auranofin triggers apoptosis through a Bax/Bak-dependent process that involves peroxiredoxin 3 oxidation . P30044 ( TrxR ) is a key selenoprotein antioxidant enzyme and a potential target for anti-cancer drugs . One potent inhibitor of TrxR is the gold ( I ) compound auranofin , which can trigger mitochondrial-dependent apoptosis pathways . The exact mechanism of apoptosis induction by auranofin is not yet clear , but there are indications that mitochondrial oxidative stress is a central event . We assessed the redox state of the peroxiredoxins ( Prxs ) in Jurkat T-lymphoma cells treated with auranofin , and found that mitochondrial Prx3 was considerably more sensitive to oxidation than the cytosolic Prx1 and 2 , indicating selective mitochondrial stress . Prx3 oxidation was detected at apoptotic doses of auranofin in several cell types , and occurred before other mitochondrial events including cytochrome c release and mitochondrial depolarisation . DB00995 was also able to sensitise U937 cells to P01375 -mediated apoptosis . DB00995 -induced apoptosis was effectively blocked by the overexpression of Bcl-2 , and Bax/Bak deficient mouse embryonic fibroblasts were also resistant to apoptosis , indicating a central role for the pro-apoptotic proteins of this family in auranofin-triggered apoptosis . DB00995 exposure inhibited the proliferation of apoptosis-resistant cells , and at higher doses of auranofin could cause cell death through necrosis . We conclude that auranofin induces apoptosis in cells through a Bax/Bak-dependent mechanism associated with selective disruption of mitochondrial redox homeostasis in conjunction with oxidation of Prx3 . Levels of angiopoietins 1 and 2 in induced sputum supernatant in patients with P48444 . Pathological features of chronic obstructive pulmonary disease ( P48444 ) include lung vascular remodeling and angiogenesis . Q15389 ( Ang-1 ) , is an essential mediator of angiogenesis by establishing vascular integrity , whereas angiopoietin-2 ( Ang-2 ) acts as its natural inhibitor . We determined the levels of angiopoietins in sputum supernatants of patients with P48444 and investigated their possible association with mediators and cells involved in the inflammatory and remodeling process . Fifty-nine patients with P48444 , 25 healthy smokers and 20 healthy non-smokers were studied . All subjects underwent lung function tests , sputum induction for cell count identification and Ang-1 , Ang-2 , P15692 , TGF-β1 , P08253 , LTB4 , P10145 , albumin measurement in sputum supernatants . Airway vascular permeability ( AVP ) index was also assessed . Ang-2 levels were significantly higher in patients with P48444 compared to healthy smokers and healthy non-smokers [ median , interquartile ranges pg/ml , 267 ( 147-367 ) vs. 112 ( 67-171 ) and 98 ( 95-107 ) , respectively ; p < 0.001 ] . Regression analysis showed a significant association between Ang-2 levels and AVP index , P15692 , P10145 and P08253 levels in P48444 , the strongest being with P15692 . Our results indicate that induced sputum Ang-2 levels are higher in P48444 compared to healthy smokers and healthy non-smokers . Moreover , Ang-2 is associated with AVP , P10145 , P08253 , and P15692 , indicating a possible role for Ang-2 in the pathogenesis of the disease .

MH_train_1543

MH_train_1543

MH_train_1543

interacts_with DB01418?

multiple_choice

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 . 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 . 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 . 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 . 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 . Study of folate receptor genes in nonsyndromic familial and sporadic cleft lip with or without cleft palate cases . DB00158 receptor family members ( FOLRs ) mediate the delivery of 5-methyltetrahydrofolate to the interior of , out of within , or between cells in a process known as potocytosis . Three FOLRs and a pseudogene map to 11q13.4 . The aim of this study was to verify whether FOLRs could be responsible for the onset of nonsyndromic cleft lip with or without cleft palate ( CL/P ) . Linkage and linkage disequilibrium between genetic markers and disorder were analyzed . Patients and their mothers from 71 familial CL/P pedigrees and 75 sporadic cases from Italian population were investigated by PCR-SSCP analysis . Data from mutation scanning allowed us to find only a silent mutation in P15328 present in a mother and her child . Our findings do not support P15328 and P14207 genes in the onset of CL/P . DB05258 /beta promotes cell survival by activating nuclear factor kappa B through phosphatidylinositol 3-kinase and Akt . Interferons ( IFNs ) play critical roles in host defense by modulating gene expression via activation of signal transducer and activator of transcription ( P35610 ) factors . IFN-alpha/beta also activates another transcription factor , nuclear factor kappaB ( NF-kappaB ) , which protects cells against apoptotic stimuli . NF-kappaB activation requires the IFN-dependent association of P40763 with the P17181 chain of the IFN receptor . IFN-dependent NF-kappaB activation involves the sequential activation of a serine kinase cascade involving phosphatidylinositol 3-kinase ( PI-3K ) and Akt . Whereas constitutively active PI-3K and Akt induce NF-kappaB activation , Ly294002 ( a PI-3K inhibitor ) , dominant-negative PI-3K , and kinase-dead Akt block IFN-dependent NF-kappaB activation . Moreover , dominant-negative PI-3K blocks IFN-promoted degradation of kappaBox alpha . Ly294002 , a dominant-negative PI-3K construct , and kinase-dead Akt block IFN-promoted cell survival , enhancing apoptotic cell death . Therefore , P40763 , PI-3K , and Akt are components of an IFN signaling pathway that promotes cell survival through NF-kappaB activation . 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 . 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 . 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 . P00747 activator in granulocyte-macrophage- P04141 transgenic mice . The pattern of expression of urokinase type plasminogen activator ( PA ) in granulocyte-macrophage- P04141 transgenic mice and their normal littermates was studied using RNAse protection assays and a plasminogen-dependent fibrinolytic assay for PA . DB00013 type PA mRNA was expressed at a high level in transgenic peritoneal cells and at a lower level in transgenic eye tissue and spleen , but not in equivalent tissue from the normal mice . Enzymically active PA was detectable in protein extracts from peritoneal cells taken from transgenic mice of less than 8 wk of age ( young mice ) but not from normals . Paradoxically , extracts from transgenic mice of more than 12 wk of age ( old mice ) showed little detectable PA activity despite continuing transcription in some mice of this age . The production of PA by peritoneal cells may be responsible for the spontaneous i.p. bleeding which is a feature of the transgenic mice and production in other tissues may help explain the local pathologic changes . 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 . 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 . c-Fes tyrosine kinase binds to and activates P40763 after granulocyte-macrophage colony-stimulating factor stimulation . Granulocyte-macrophage colony stimulating factor ( GM- P04141 ) induces proliferation and maturation of myeloid progenitor cells and also activates neutrophils . In order to investigate the pleiotropic effects of GM- P04141 stimulation , we examined the signaling pathways of protein tyrosine kinases ( PTKs ) and signal transducers and activators of transcription ( STATs ) in GM- P04141 -dependent proliferation of leukemia cells . Using TF-1 , a GM- P04141 -dependent human erythroleukemia cell line , we found that GM- P04141 enhanced DNA-binding and tyrosine phosphorylation of P40763 . GM- P04141 receptor ( GM-CSFR ) and c-Fes tyrosine kinase were also activated upon GM- P04141 stimulation . Furthermore , c-Fes formed a complex with P40763 . Experiments using a c-Fes mutant that lacked tyrosine kinase activity revealed that the activation of P40763 is kinase-dependent , but that the c-Fes- P40763 interaction is not affected by c-Fes tyrosine kinase activity . The results suggest that P40763 is activated by c-Fes tyrosine kinase through direct interaction during hematopoietic cell proliferation induced by GM- P04141 . Clinical pharmacology of serotonin-altering medications for decreasing alcohol consumption . Variations in serotonin neurotransmission influence alcohol consumption ( AC ) . Levels of 5-HT and metabolites are low in some brain regions of alcohol preferring rats and in P04141 of alcoholics . Pharmacological treatments which enhance serotonergic neurotransmission ( uptake inhibitors , releasers , agonists ) consistently reduce AC in rats . Serotonin uptake inhibitors ( SUI ; e.g. , citalopram , fluoxetine ) have been studied extensively in humans . In several double-blind randomized , placebo-controlled clinical trials , SUI have consistently decreased AC by averages of 15 % to 20 % in nondepressed mildly/moderately dependent alcoholics who received no other treatment . Effects were dose-dependent and not related to side effects ( few and mild ) or changes in anxiety or depression ( not observed ) . SUI also decreased desire to drink and liking for alcohol , thus suggesting a mechanism for effects . Other drugs acting on the 5-HT system have been tested in humans , but results are difficult to interpret . For example , buspirone , a P08908 receptor partial agonist , reduced anxiety and alcohol craving , but not AC ; a 5-HT partial agonist , m-CPP , increased alcohol craving in abstinent alcoholics ; modest reductions in AC were observed with a 5- Q9H205 antagonist , ondansetron ( 0.5 mg/day , but not 4 mg/day ) . The therapeutic potentials of these medications are being studied . For example , SUI effects on AC were enhanced by a brief psychosocial intervention . Since SUI decrease urge to drink , they may be suitable pharmacological adjuncts in relapse prevention strategies . SUI and other serotonin-altering medications are promising new neuropharmacological treatments for reducing AC . 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 . 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 . Low levels of insulin-like growth factor-I in cerebrospinal fluid in children with autism . Autism is a behaviourally defined syndrome characterized by disturbances of social interaction and communication and restrictions of behaviour patterns and imagination . The pathogenesis of autism is unknown but it is suspected that a number of genetic factors may be involved . Neurotrophic factors such as insulin-like growth factor-I ( P05019 ) play a role in early brain development . The aim of this study was to determine whether P05019 levels might be associated with the development of autism . P05019 levels were measured in the P04141 of 11 children with autism ( 4 females , 7 males ; mean age 3.8 years , SD 1.1 ) using a sensitive radioimmunoassay method and compared with levels in 11 control participants ( 6 females , 5 males ; mean age 3.8 years ) . Levels of P05019 in the P04141 were statistically significantly lower in the children with autism than in the control children ( p=0.03 ) . P05019 may play a role in pathogenetic mechanisms of autism and the role of neurotrophic factors in autism and other neurodevelopmental diseases should be studied further . 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 . Fas/CD95-induced chemokines can serve as " find-me " signals for apoptotic cells . Apoptosis is commonly thought to represent an immunologically silent or even anti-inflammatory mode of cell death , resulting in cell clearance in the absence of explicit activation of the immune system . However , here we show that Fas/CD95-induced apoptosis is associated with the production of an array of cytokines and chemokines , including P05231 , P10145 , P09341 , P13500 , and P04141 . Fas-induced production of P13500 and P10145 promoted chemotaxis of phagocytes toward apoptotic cells , suggesting that these factors serve as " find-me " signals in this context . We also show that Q13546 and IAPs are required for optimal production of cytokines and chemokines in response to Fas receptor stimulation . Consequently , a synthetic IAP antagonist potently suppressed Fas-dependent expression of multiple proinflammatory mediators and inhibited Fas-induced chemotaxis . Thus , in addition to provoking apoptosis , Fas receptor stimulation can trigger the secretion of chemotactic factors and other immunologically active proteins that can influence immune responsiveness toward dying cells .

MH_train_1544

MH_train_1544

MH_train_1544

interacts_with DB00015?

multiple_choice

P22309 *28 is associated with greater decrease in serum K⁺ levels following oral intake of procaterol . BACKGROUND AND OBJECTIVE : DB01366 is a potent β2-agonist frequently used for the management of asthma and chronic obstructive pulmonary disease . The efficacy and adverse effects of β2-agonists are heterogeneous in individual patients , which may be partly caused by genetic variations in metabolizing enzymes and receptor molecules . The present study was designed to analyze the relationship between gene polymorphisms and physiological effects of procaterol in healthy subjects . METHODS : Ninety-two non-smoking healthy volunteers were given 1 µg/kg body weight ( max 50 µg ) of procaterol as a dry syrup preparation , and the serum concentrations of procaterol , serum K(+) , and the physical responses were monitored for 240 min . We genotyped β2-adrenergic receptor ( P07550 ) ( Arg16Gly and Gln27Glu ) , cytochrome P450 3A4 ( rs2246709 , rs4646437 ) , and uridine diphosphate glucuronosyltransferase 1A1 ( P22309 ) ( rs4148323 [ allele A , *6 ] , rs12479045 , rs4148328 , rs4663971 , rs12052787 , rs4148329 , A (TA)6/7 TAA [ seven-repeat allele , *28 ] ) . DB01366 concentrations in serum were measured by liquid chromatography-tandem mass spectrometry . RESULTS : No gene polymorphisms affected serum procaterol concentrations . Meanwhile , overall serum K(+) level changes were significantly lower in carriers of P22309 *28 than in non-carriers after correcting for strong effects of serum procaterol concentrations and baseline K(+) levels . No other polymorphisms were associated with serum K(+) levels . None of polymorphisms of P07550 were associated with any physical responses . CONCLUSION : The present study indicates that significant hypokalemia may occur in carriers of P22309 *28 by systemic administration of procaterol and potentially by other β2-agonists metabolized in the liver . [ Association study of renin-angiotensin system genes and hemostasis system genes with ischemic stroke among Russians of Central Russia ] . The analysis of alleles and genotypes frequencies of 14 SNP in genes of rennin-angiotensin system ( REN , AGT , P30556 , P50052 , P30411 , P07550 ) and hemostasis system ( P02675 , F2 , P12259 , P08709 , P05106 , P05121 , P42898 ) , as well as P12821 insertion-deletion polymorphism in patients with stroke comparing to healthy controls matched by age , sex and ethnicity has been carried out . The genotyping procedure included the amplification of selected gene sequences following by hybridization of fluorescently labeled fragments with SNP-specific DNA probes . The analysis of allele frequencies of each gene separately revealed no statistically significant differences between groups of patients with stroke and healthy donors . Also the complex study has been performed to estimate the contribution of rennin-angiotensin system and hemostasis system genes to the genetic susceptibility to ischemic stroke among Russians from Central Russia using method MDR ( Multifactor Dimensionality Reduction ) . The combination with increased risk for development of ischemic stroke was presented by complex genotype P02675 G/- x P12821 I/- x P42898 C/- x P05121 5G/5G ( p = 0.03 , OR = 2.4 , 95 % CI 1.1-5.3 ) , which frequency was statistically significant higher in patients with stroke compared to healthy control . Inhibition of p38alpha MAPK disrupts the pathological loop of proinflammatory factor production in the myelodysplastic syndrome bone marrow microenvironment . Myelodysplastic syndromes ( P43034 ) are common causes of ineffective hematopoiesis and cytopenias in the elderly . Various myelosuppressive and proinflammatory cytokines have been implicated in the high rates of apoptosis and hematopoietic suppression seen in P43034 . We have previously shown that p38 MAPK is overactivated in P43034 hematopoietic progenitors , which led to current clinical studies of the selective p38alpha inhibitor , DB05412 , in this disease . We now demonstrate that the myelosuppressive cytokines TNFalpha and IL-1beta are secreted by bone marrow ( BM ) cells in a p38 MAPK-dependent manner . Their secretion is stimulated by paracrine interactions between BM stromal and mononuclear cells and cytokine induction correlates with P28906 + stem cell apoptosis in an inflammation-simulated in vitro bone marrow microenvironment . Treatment with DB05412 inhibits P01375 secretion in primary P43034 bone marrow cells and protects cytogenetically normal progenitors from apoptosis ex vivo . Furthermore , p38 inhibition diminishes the expression of TNFalpha or IL-1beta-induced proinflammatory chemokines in BM stromal cells . These data indicate that p38 inhibition has anti-inflammatory effects on the bone marrow microenvironment that complements its cytoprotective effect on progenitor survival . These findings support clinical investigation of p38alpha as a potential therapeutic target in P43034 and other related diseases characterised by inflammatory bone marrow failure . Protein assay for heme oxygenase-1 ( P09601 ) induced by chemicals in HepG2 cells . Levels of heme oxygenase-1 ( P09601 ) , a stress response protein , were measured to examine oxidative stress induced by several chemicals in HepG2 cells with and without S9mix using an ELISA . CdCl(2) , heme , and diclofenac sodium salt ( diclofenac ) were used as inducers of P09601 . Acetaminophen ( P08697 ) and cyclophosphamide ( CP ) were used as oxidative stress inducers . Stannic mesoporphyrin ( DB04912 ) was used as an inhibitor of HO activity . Cytotoxicity was determined , and P09601 levels were measured in HepG2 cells exposed to chemicals other than CP with non-metabolic activation without S9mix , and to diclofenac , P08697 and CP with metabolic activation with S9mix . P09601 levels were increased by CdCl(2) ( 7.5 microM ) , heme ( 10 , 100 microM ) , and stannic mesoporphyrin ( DB04912 ) ( 10 microM ) , but were not changed by P08697 , and were decreased by diclofenac . P09601 levels were increased by diclofenac ( 300 microM ) , and CP ( 36 microM ) , but were unaffected by P08697 because of low sensitivity in HepG2 cells . The induction of P09601 expression was first observed in cultured HepG2 cells treated with CP under conditions involving metabolic activation . These results showed the measurement of P09601 protein levels in this system is useful when assessing oxidative stress as a tool for detecting drug toxicity . 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 . 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 . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . A pegylated growth hormone receptor antagonist , pegvisomant , does not enter the brain in humans . BACKGROUND : GH receptors exist in the hippocampus , cerebral cortex , and hypothalamus , possibly influencing mood , cortical blood flow , and neuronal growth and mediating negative feedback . RATIONALE : DB00082 is a recombinant mutated GH molecule with high affinity , but little or no activating capability , for the P10912 . It is used clinically as a GH antagonist . HYPOTHESIS : Systemic pegvisomant enters brain interstitial fluid via putative choroid-plexus GH receptors , thereby allowing for antagonism of central actions of GH . SUBJECTS AND LOCATION : Six adults requiring a cerebrospinal fluid ( P04141 ) examination for nonneoplastic and noninflammatory syndromes participated at a tertiary medical center . METHODS : Direct assays were conducted of serum and P04141 pegvisomant concentrations 18-24 h after sc injection of pegvisomant ( 20 mg ) . OUTCOMES : Median ( range ) concentrations of pegvisomant in serum were 215 ( 74-539 ) microg/liter and in P04141 0.035 ( 0.010-0.28 ) microg/liter ( P=0.016 ) . P04141 drug levels were indistinguishable from assay threshold . Corresponding GH values were 0.29 ( 0.010-1.3 ) in serum and 0.075 microg/liter ( 0.01-0.13 ) in P04141 . The geometric mean ratios of serum/ P04141 pegvisomant and GH concentrations were 5116:1 and 3.5:1 , respectively , thus defining a more than 1400-fold difference between mutated and natural GH . CONCLUSIONS : Based upon P04141 measurements , a pegylated GH-receptor antagonist does not cross the human blood-brain barrier , thereby sparing inhibition of central nervous system GH actions . Thus , the capability of this antagonist to stimulate GH secretion predominantly reflects its actions outside the blood-brain barrier , such as via the median eminence and/or via suppression of P05019 concentrations . JAK/ P40763 signaling is required for TGF-β-induced epithelial-mesenchymal transition in lung cancer cells . Epithelial-mesenchymal transition ( EMT ) , a key step in the early stages of cancer metastasis , is orchestrated by several signaling pathways , including P05231 /JAK/ P40763 and TGF-β/Smad signaling . However , an association between the two signaling pathways during the EMT process is largely unknown . Here , we focused on lung cancer and demonstrated that TGF-β1 induced the phosphorylation of P84022 ( p- P84022 ) , upregulation of Snail , a fibroblast-like morphology , and downregulation of P12830 as well as upregulation of vimentin in lung cancer cell lines . SIS3 ( an inhibitor of P84022 ) suppressed TGF-β1-induced activation of P84022 , upregulation of Snail and the EMT process . Importantly , the O60674 / P40763 -specific inhibitor AG490 blocked Stat3 phosphorylation , resulting in attenuated levels of TGF-β1-induced p- P84022 , Snail , P08253 , and Smad-mediated P05121 promoter reporter gene activity in A549 and H1650 cells . Subsequently , AG490 inhibited TGF-β-induced cell migration and invasion . Moreover , exogenous P05231 treatment stimulated Stat3 activation , enhanced TGF-β-induced expression of p- P84022 and Snail , aggravated the EMT process , and increased lung cancer cell migration and invasion induced by TGF-β1 . Our findings show that the JAK/ P40763 pathway is required for TGF-β-induced EMT and cancer cell migration and invasion via upregulation of the expression of p- P84022 and Snail , and the P05231 /JAK/ P40763 and TGF-β/Smad signaling synergistically enhance EMT in lung carcinomas . The present study suggests a novel rationale for inhibiting cancer metastasis using anti- P05231 /JAK/ P40763 and anti-TGF-β/Smad therapeutic strategies . Association studies between the plasmin genes and late-onset Alzheimer 's disease . The plasmin system is involved in the degradation of Abeta peptides , the accumulation of which in brain is a hallmark of Alzheimer 's disease ( AD ) . In a North European case-control AD dataset we studied 14 common variations in the P00747 , P05121 , P00750 and P08697 genes encoding components of the plasmin system . Among the four polymorphisms in the P00750 , P05121 and P08697 genes showing nominally significant evidence for an association with AD ( allele p-value=0.01-0.00003 ) the strongest association was detected for the deletion allele in the Alu-repeat region of the P00750 gene . However , none of these positive results were confirmed in follow-up studies using an independent Canadian case-control cohort and two familial AD datasets of North European and Caribbean Hispanic origin . Thus , the current survey does not support the notion that common polymorphisms in the plasmin genes influence the development of AD . Efficacy and safety of etoricoxib in the treatment of osteoarthritis . Osteoarthritis is a progressive disease that affects millions of people worldwide , but for which there are no curative options and indeed a limited number of medical treatment options . The American College of Rheumatology recommendations suggest administering either a traditional NSAID or a P35354 -selective inhibitor for pain relief . Traditional NSAIDs , such as naproxen , may have a higher risk of gastrointestinal ( GI ) events , while P35354 -selective inhibitors may have a higher risk of thrombotic cardiovascular ( CV ) events ( with traditional NSAIDs and P35354 -selective agents appearing to have a similar CV risk ) . DB01628 , introduced in 2002 , has been approved in over 60 countries worldwide for osteoarthritis . Large-scale studies addressing the efficacy , GI tolerability and potential for CV events with etoricoxib have now been published . Several patient types appear to benefit from etoricoxib , including those with CV risk factors and those requiring gastroprotective agents . In patients with CV risk factors , the benefits and risks of all NSAIDs should be weighed carefully in each patient , balancing the potential risks of treatment against the potential relief for pain and disability . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . 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 . Gene expression profiling of LPS-stimulated murine macrophages and role of the NF-kappaB and PI3K/ P42345 signaling pathways . Lipopolysaccharide ( LPS ) , a major component of the outer membrane of Gram-negative bacteria , activates a broad spectrum of signaling pathways in immune cells . In this article , RAW264.7 cells have been stimulated for 4 h with 1 microg/mL of LPS in the presence or not of specific inhibitors of the NF-kappaB pathway ( BAY 11-7082 ) and the PI3K pathway ( LY294002 ) . Gene expression profiles were characterized using the DNA microarray " Dual Chip Mouse Inflammation. " This array monitors the expression of 233 genes encoding proteins playing a role in inflammation . Both signaling pathways exert an important role in the response to LPS , but they are not completely overlapping . For example , genes encoding the Q15004 receptor , P05121 , PlA2 ( group V ) , P35225 receptor ( alpha2 ) , and P30793 , were upregulated after LPS treatment , but this upregulation was counteracted by LY294002 . The same was observed for BAY 11-7082 : genes encoding the kit ligand , O60603 , or P25942 were mainly under the control of NF-kappaB . NF-kappaB plays an important role in the macrophage response to LPS , but we have also shown that the PI3K pathway partially contributes to it . Further experiments with the specific inhibitor of P42345 ( rapamycin ) will provide more information on the specific contribution of the PI3K/ P42345 pathway in the inflammatory response in LPS-stimulated macrophages . 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 . 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 . The Multifaceted Clinical Readouts of Platelet Inhibition by Low-Dose DB00945 . Inactivation of platelet cyclooxygenase ( P36551 ) -1 by low-dose aspirin leads to long-lasting suppression of thromboxane ( TX ) A2 production and TXA2-mediated platelet activation and aggregation . This effect is necessary and sufficient to explain aspirin 's unique ( among other P23219 inhibitors ) effectiveness in preventing atherothrombosis , as well as its shared ( with other antiplatelet agents ) bleeding liability . However , different mechanisms of action have been suggested to explain other beneficial effects of aspirin , such as prevention of venous thromboembolism , chemoprevention of colorectal ( and other ) cancers , and reduced risk of dementia . These mechanisms include acetylation of other proteins in blood coagulation , inhibition of P35354 activity , and other P36551 -independent mechanisms . The intent of this review is to develop the concept that the multifaceted therapeutic effects of low-dose aspirin may reflect pleiotropic consequences of platelet inhibition on pathophysiological tissue repair processes . Furthermore , the clinical implications of this concept will be discussed in terms of current clinical practice and future research .

MH_train_1545

MH_train_1545

MH_train_1545

interacts_with DB01151?

multiple_choice

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 . P12004 D1-Cdk4 controls glucose metabolism independently of cell cycle progression . P01308 constitutes a principal evolutionarily conserved hormonal axis for maintaining glucose homeostasis ; dysregulation of this axis causes diabetes . P20142 -1α ( peroxisome-proliferator-activated receptor-γ coactivator-1α ) links insulin signalling to the expression of glucose and lipid metabolic genes . The histone acetyltransferase Q92830 ( general control non-repressed protein 5 ) acetylates P20142 -1α and suppresses its transcriptional activity , whereas sirtuin 1 deacetylates and activates P20142 -1α . Although insulin is a mitogenic signal in proliferative cells , whether components of the cell cycle machinery contribute to its metabolic action is poorly understood . Here we report that in mice insulin activates cyclin D1-cyclin-dependent kinase 4 ( Cdk4 ) , which , in turn , increases Q92830 acetyltransferase activity and suppresses hepatic glucose production independently of cell cycle progression . Through a cell-based high-throughput chemical screen , we identify a Cdk4 inhibitor that potently decreases P20142 -1α acetylation . P01308 /GSK-3β ( glycogen synthase kinase 3-beta ) signalling induces cyclin D1 protein stability by sequestering cyclin D1 in the nucleus . In parallel , dietary amino acids increase hepatic cyclin D1 messenger RNA transcripts . Activated cyclin D1-Cdk4 kinase phosphorylates and activates Q92830 , which then acetylates and inhibits P20142 -1α activity on gluconeogenic genes . Loss of hepatic cyclin D1 results in increased gluconeogenesis and hyperglycaemia . In diabetic models , cyclin D1-Cdk4 is chronically elevated and refractory to fasting/feeding transitions ; nevertheless further activation of this kinase normalizes glycaemia . Our findings show that insulin uses components of the cell cycle machinery in post-mitotic cells to control glucose homeostasis independently of cell division . Long-term treatment with the antidepressants fluoxetine and desipramine potentiates endocrine responses to the serotonin agonists 6-chloro-2-[1-piperazinyl]-pyrazine ( MK-212 ) and (+-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl ( DOI ) . Various endocrine responses to 5-hydroxytryptamine ( serotonin , 5-HT ) agonists were used to assess serotonergic receptor function after chronic treatment with the antidepressants fluoxetine ( 10 mg/kg ) , a 5-HT uptake blocker and the norepinephrine uptake blocker desipramine ( DB01151 , 5 mg/kg ) . Both were injected ( i.p. ) once a day for 21 days . DOI ( P28335 /2 agonist , 0-5 mg/kg i.p. ) and 6-chloro-2-[1-piperazinyl]-pyrazine ( MK-212 ) ( less selective , but predominantly a P28335 agonist , 0-20 mg/kg i.p. ) were administered 18 hr after the final antidepressant injection and 30 min before decapitation . Chronic treatment with both fluoxetine and DB01151 produced a potentiation in most hormone responses to the 5-HT agonists (+-)-1-(2,5-dimethoxy-4-iodophenyl)-2-amino-propane HCl ( DOI ) and MK-212 , although there were several differences in individual hormone responses to the two 5-HT agonists . DB00472 and DB01151 potentiated the MK-212- and DOI-induced increase of plasma oxytocin levels and potentiated the effect of DOI on plasma adrenocorticotropic hormone ( corticotropin ) and prolactin levels . In contrast , the effect of the high dose of MK-212 on plasma prolactin concentration was reduced by both antidepressants . Only MK-212 increased vasopressin levels and this effect was potentiated by fluoxetine , but not by DB01151 . DB00472 also significantly increased the resting level of plasma vasopressin . DB01151 potentiated the effect of MK-212 on plasma renin concentration . Pretreatment with fluoxetine significantly increased ( 38 % ) the Bmax for the P28335 /2 agonist sites ( [125I]DOI ) in the hypothalamus. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO:fat:protein = 12:59:28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO:fat:protein = 56:24:20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n-6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4n-6 ) , while its biosynthetic metabolic intermediates were decreased . The n-6/n-3 and arachidonic/eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1n-7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 , P05231 , P10145 , P13500 , P16581 , I- P62158 , and P05121 . Increased 20:4n-6 and the ratios of 20:4n-6/20:5n-3 and n-6/n-3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet . 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 . Placental histomorphology in unexplained foetal loss with thrombophilia . BACKGROUND & OBJECTIVES : Acquired and genetic thrombotic conditions , both organ and non organ specific , are associated with increased foetal wastage . This study was carried out to examine the placenta from women with abnormal pregnancies and a history of unexplained foetal loss , and to associate with maternal thrombophilia status . METHODS : Placentas from eight women with history of unexplained foetal loss were analyzed for histopathological characteristics . All the women were simultaneously screened for the common acquired and genetic thrombophilia markers i.e. , lupus anticoagulants ( LA ) , IgG / IgM antibodies for anticardiolipin ( ACA ) , beta2 glycoprotein 1 ( beta2GPI ) and annexin V , protein C ( PC ) , protein S ( PS ) , antithrombin III ( AT III ) , factor V Leiden ( FVL ) mutation , prothrombin ( PT ) gene G20210A , methylene DB00116 reductase ( P42898 ) C 677T , endothelial protein C receptor ( Q9UNN8 ) 23 bp insertion and plasminogen activator inhibitor ( P05121 4G/5G ) polymorphisms RESULTS : Six of eight women were positive for one or more thrombophilia markers . The placenta in all the cases except one , showed the characteristic features of infarct fibrin deposition and calcification . Among two women who were negative for thrombophilia , one showed clear evidence of thrombus in the placental sections while the other did not show any characteristic infarcts in the placental sections . INTERPRETATION & CONCLUSION : Our findings showed that the histopathological examination of the placentas confirmed thrombophilia as the aetiological cause of thrombosis in 6 of the 8 women . The presence of thrombus in a negative thrombophilia woman suggests yet unidentified thrombophilia markers or probably non-haemostatic factors causing thrombosis . 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 . 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 . Effects of a novel P08908 receptor agonist , E4424 , on gastric adherent mucus levels following restraint stress in rats . Several novel arylpiperazine serotonin 1A receptor agonists , developed as anxiolytics , have antisecretory and gastroprotective effects in rats . E4424 ( 2-¿4-[4-(4-chloropyrazol-1-yl)butyl]-1-piperazinyl ¿pyrimidine ; DB04970 dihydrochloride ) , has potent anti-gastric secretory and antiulcer effects . Preliminary data indicated an enhancing effect of E4424 on gastric mucus that may underlie its gastroprotective actions . We therefore tested the effects of acute and chronic administration of E4424 and of a reference P08908 receptor agonist , 8-OHDPAT [ 8-hydroxy-2-(di-n-propylamino)tetralin ] , on gastric mucus levels in rats subjected to cold-restraint stress , a procedure associated with depletion of gastric mucus and the development of mucosal injury . Acute oral administration of E4424 increased adherent mucus levels by 12 % , 11 % , and 13 % , relative to controls . Chronic E4424 significantly increased gastric mucus relative to controls ( 69 % increase ) . Acute oral treatment with 8-OHDPAT did not affect gastric mucus level . Acute intraperitoneal 8-OHDPAT slightly increased mucus levels . Chronic twice per day 8-OHDPAT did not affect mucus levels ; however , chronic once per day treatment with 8-OHDPAT significantly elevated gastric mucus levels at the highest doses used . For E4424 , there is a strong correlation between reduction of gastric mucosal injury and increase in gastric mucus level , suggesting that the action of E4424 on glandular mucus levels is an important mechanism underlying its gastroprotective effects . Randomised clinical trial : effects of monotherapy with DB05070 , a P41594 inhibitor , on symptoms and reflux events in patients with gastro-oesophageal reflux disease . BACKGROUND : DB05070 , a metabotropic glutamate receptor 5 ( P41594 ) negative allosteric modulator , has been shown to reduce gastro-oesophageal reflux events and oesophageal acid exposure in patients with gastro-oesophageal reflux disease ( GERD ) and healthy subjects . AIM : To evaluate the effects of DB05070 monotherapy for 2 weeks on symptom control in patients with GERD . METHODS : This was a double-blind , placebo-controlled , multi-centre trial in GERD patients who were responders to proton pump inhibitors ( PPIs ) . Following PPIs withdrawal , a 2-week baseline washout period was followed by 2-week treatment with either DB05070 120 mg or placebo b.d . The primary clinical efficacy endpoint was the number of GERD symptom-free days in treatment week 2 compared with the last 7 days of baseline . The effect on reflux events using 24-h impedance-pH monitoring was also determined in a subset of 24 patients . RESULTS : The full analysis set comprised 103 patients DB05070 ( N= 50 ) , Placebo ( N=53 ) . In treatment week 2 , DB05070 significantly increased GERD symptom-free days ( P=0.045 ) and heartburn-free days ( P=0.037 ) , reduced antacid use ( P=0.017 ) , improved total symptom score ( P=0.048 ) including subscale heartburn/regurgitation ( P=0.007 ) and sleep disturbance because of GERD ( P= 0.022 ) . DB05070 significantly reduced total ( P=0.034 ) and acidic reflux events ( P=0.003 ) . DB05070 was well tolerated . Most common adverse events for DB05070 were mild to moderate dizziness 16 % and vertigo 12 % ( placebo 4 % and 2 % ) . CONCLUSIONS : Inhibition of P41594 with DB05070 monotherapy reduces reflux events and improves symptoms in GERD patients . This mechanism has promise for the management of GERD . 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 . 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 . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . Comparative molecular profiling of the PPARα/γ activator aleglitazar : Q07869 selectivity , activity and interaction with cofactors . Peroxisome proliferator-activated receptors ( PPARs ) are a family of nuclear hormone receptors that control the expression of genes involved in a variety of physiologic processes , through heterodimerization with retinoid X receptor and complex formation with various cofactors . Drugs or treatment regimens that combine the beneficial effects of PPARα and γ agonism present an attractive therapeutic strategy to reduce cardiovascular risk factors . DB08915 is a dual PPARα/γ agonist currently in phase III clinical development for the treatment of patients with type 2 diabetes mellitus who recently experienced an acute coronary event . The potency and efficacy of aleglitazar was evaluated in a head-to-head comparison with other PPARα , γ and δ ligands . A comprehensive , 12-concentration dose-response analysis using a cell-based assay showed aleglitazar to be highly potent , with EC(50) values of 5 nM and 9 nM for PPARα and PPARγ , respectively . Cofactor recruitment profiles confirmed that aleglitazar is a potent and balanced activator of PPARα and γ . The efficacy and potency of aleglitazar are discussed in relation to other dual PPARα/γ agonists , in context with the published X-ray crystal structures of both PPARα and γ . A continuous , nonradioactive assay for histone acetyltransferases . Histone acetyltransferases ( HATs ) catalyze the acetyl-group transfer from acetyl- DB01992 to the epsilon-amino group of specific lysine residues within core histone proteins . HATs and other chromatin-remodeling enzymes have been recently shown to regulate gene activation within specific loci . To facilitate mechanistic studies , we have developed two continuous , nonradioactive assays for the prototypical Q92830 O60235 . The DB01992 generated in the O60235 reactions was continuously measured by using a coupled enzyme system with either alpha-ketoglutarate dehydrogenase or pyruvate dehydrogenase . The DB01992 -dependent oxidation of alpha-ketoglutarate or pyruvate is accompanied by the reduction of NAD to DB00157 , which was measured spectrophotometrically at 340 nm . The steady-state rate constants with substrates acetyl- DB01992 and a synthetic peptide ( corresponding to the first 20 amino acids of H3 histone ) were determined . The resulting rate constants were not significantly different between the two coupled assays , providing strong validation of these methods . Rate constants were also determined using the commonly employed radioactive filter-binding assay and compared . The 1.5- to 5-fold lower values obtained in the radioactive end-point assay are discussed in terms of the technical problems and limitations of this assay . The coupled assays should be widely applicable since the production of DB01992 is common to all O60235 enzymes , regardless of protein substrate . 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 . 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 . 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 . Individual cerebellar Purkinje cells express different cGMP phosphodiesterases ( PDEs ) : in vivo phosphorylation of cGMP-specific PDE ( O76074 ) as an indicator of cGMP-dependent protein kinase ( PKG ) activation . The nitric oxide ( NO ) -cGMP pathway has been implicated as playing a crucial role in the induction of cerebellar long-term depression ( LTD ) . The amplitude and duration of the cGMP signal is controlled by cyclic nucleotide phosphodiesterases ( PDEs ) . Here we identify O76074 and Q01064 as the two major cGMP-hydrolyzing PDEs specifically and differentially expressed in the Purkinje neurons of mouse cerebellum . O76074 was found in all Purkinje neurons , whereas Q01064 was detected only in a subset of these cells , suggesting that individual Purkinje cells may differentially regulate cGMP , depending on the PDE isozymes expressed . Although expression of guanylate cyclase and/or cGMP-dependent protein kinase ( PKG ) in Purkinje cells have been reported , neither cGMP accumulation nor PKG activation in these cells in vivo has been demonstrated . To determine if changes in PKG activation and O76074 regulation occur in vivo we have examined the phosphorylation of O76074 in mouse cerebellar Purkinje cells by immunocytochemistry and Western blot analyses using a phosphospecific O76074 antibody . Injection of sodium nitroprusside or selective PKG activators into the lateral ventricle of mouse brain induced O76074 phosphorylation in vivo , but was completely missing in Purkinje cell-specific PKG I knock-out mice . In cerebellar slices , treatment with sildenafil or DB07954 led to different levels of phospho- O76074 accumulation and activation of O76074 . These results suggest that phosphorylation of O76074 in Purkinje neurons after cGMP-PKG activation performs a critical role in the termination of the cGMP signal during LTD progression ; moreover , O76074 phosphorylation may be used as an in vivo indicator for PKG activation . 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 . 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 . 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 . Niflumic acid and MSI-2216 reduce P01375 -induced mucin expression in human airway mucosa . BACKGROUND : Human chloride channel , calcium-activated 1 ( A8K7I4 ) has been shown to induce mucin ( MUC ) gene expression and mucus production in bronchial epithelial cells . Objective To investigate whether blocking A8K7I4 decreases mucus production . METHODS : Expression of A8K7I4 and mucus was stimulated with P01375 in human upper airway mucosal explant tissue . P98088 mRNA and mucus protein expression was blocked by inhibiting A8K7I4 by using channel blockers ( niflumic acid [ DB04552 ] and MSI-2216 ) without and with P01375 stimulation . Expression of P98088 , A8K7I4 , and P35354 mRNA was quantified by using real-time PCR . Mucus protein was assessed by periodic acid Schiff staining . Laser capture microdissection was performed to quantify A8K7I4 and P98088 mRNA expression in epithelial cells derived from mucosal explant tissue . RESULTS : P01375 significantly increased P98088 and A8K7I4 mRNA as well as mucus and A8K7I4 protein expression in the mucosal explant tissue ( P < .05 ) . Inhibition of A8K7I4 with DB04552 or MSI-2216 showed a significant dose-dependent reduction of mucus production for both blockers in the mucosal explant tissue ( P < .05 ) . P98088 mRNA was also decreased by both blockers in the whole mucosal tissue and in laser-captured mucosa epithelial cells . CONCLUSIONS : Unstimulated and P01375 -induced mucin expression could be decreased by DB04552 and MSI-2216 . Inhibiting A8K7I4 may be a potential new approach to reduce mucus overproduction . The study of genetic polymorphisms related to serotonin in Alzheimer 's disease : a new perspective in a heterogenic disorder . Genetic and environmental factors have been implicated in the development of Alzheimer 's disease ( AD ) , the most common form of dementia in the elderly . Mutations in 3 genes mapped on chromosomes 21 , 14 and 1 are related to the rare early onset forms of AD while the epsilon 4 allele of the apolipoprotein E ( P02649 ) gene ( on chromosome 19 ) is the major susceptibility locus for the most common late onset AD ( LOAD ) . Serotonin ( 5-hydroxytryptamine or 5-HT ) is a key neurotransmitter implicated in the control of mood , sleep , appetite and a variety of traits and behaviors . Recently , a polymorphism in the transcriptional control region upstream of the 5-HT transporter ( 5-HTT ) gene has been studied in several psychiatric diseases and personality traits . It has been demonstrated that the short variant(s) of this 5-HTT gene-linked polymorphic region ( 5-HTTLPR ) is associated with a different transcriptional efficiency of the 5-HTT gene promoter resulting in decreased 5-HTT expression and 5-HT uptake in lymphocytes . An increased frequency of this 5-HTTLPR short variant polymorphism in LOAD was recently reported . In addition , another common polymorphic variation in the 5- Q13049 and P28335 serotonin receptor genes previously analyzed in schizophrenic patients was associated with auditory and visual hallucinations in AD . These observations suggest that the involvement of the serotonin pathway might provide an explanation for some aspects of the affective symptoms commonly observed in AD patients . In summary , research on genetic polymorphisms related to AD and involved in receptors , transporter proteins and the enzymatic machinery of serotonin might enhance our understanding of this devastating neurodegenerative disorder . DB09302 : Q8NBP7 inhibitor for LDL cholesterol reduction . The proof of concept that proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) inhibition affects cholesterol levels was first established after the demonstration that Q8NBP7 loss-of-function mutations result in a significant drop in circulating LDL cholesterol levels . Subsequent studies revealed that Q8NBP7 binds the epidermal growth factor precursor homology domain-A on the surface LDL Receptor ( P01130 ) and directs P01130 and Q8NBP7 for lysosomal degradation . DB09302 ( also known as SAR236553/REGN727 ) is a monoclonal antibody that binds circulating Q8NBP7 and blocks its interactions with surface P01130 . DB09302 clinical trials with different doses on different administration schedules were shown to significantly reduce LDL cholesterol both as a mono-therapy and in combination with statins or ezetimibe . Although there is great potential for anti- Q8NBP7 therapies in the management of cholesterol metabolism , there is no clear evidence yet that blocking Q8NBP7 reduces cardiovascular disease outcome . This is being investigated in ongoing Phase III clinical trials with alirocumab . Genetic and environmental influences on total plasma homocysteine and coronary artery disease ( CAD ) risk among South Indians . BACKGROUND : Hyperhomocysteinemia , a documented risk factor for CAD is highly prevalent in Indians . The rationale behind the current study is to explore the genetic and environmental causes for such high prevalence as there are limited studies in this context . METHODS : A total of 108 CAD cases and 108 controls were analyzed for tHcy and 4 folate pathway genetic polymorphisms [ methylene DB00116 reductase ( P42898 ) C677T , 5-methyltetrahydrofolate homocysteine methyl transferase ( Q99707 ) A2756G , methionine synthase reductase ( Q9UBK8 ) A66G and glutamate carboxypeptidase II ( Q04609 ) C1561T ] using reverse phase HPLC and PCR-RFLP methods respectively . RESULTS : P42898 677T , Q9UBK8 66A , Q04609 1561T , male gender , alcohol intake , smoking , diabetes , creatinine and hypertension were found to influence tHcy . After controlling for confounding factors , Hyperhomocysteinemia and two of its genetic determinants i.e. P42898 C677T [ OR : 1.96 , 95 % CI : 1.06-3.61 ] and P04001 II C1561T [ OR : 2.09 , 95 % CI : 1.09-3.97 ] were found to be associated with risk for CAD . Significant epistatic interactions were observed between P42898 677T/ Q99707 2756G and P04001 II 1561T/ Q9UBK8 66G . DB00898 intake in subjects with Q99707 2756G allele was found to inflate the risk for CAD [ OR : 4.15 , 95 % CI : 1.35-12.69 ] . CONCLUSION : Hyperhomocysteinemia , C677T P42898 and C1561T Q04609 are risk factors for CAD . Potential gene -- gene and gene -- environment interactions indicate the need for multi-variate analyses for risk prediction . Functional significance of subtypes of 5-HT receptors in the rat spinal reflex pathway . 1. The functional significance of subtypes of 5-hydroxytryptamine ( 5-HT ) receptors was studied in the rat spinal reflex pathway . 2 . Ketanserin had no effect on the mono- ( Q9UBK8 ) or polysynaptic reflex ( PSR ) in spinal rats , but decreased the PSR in intact rats . 3 . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and 5-methoxy-N,N-dimethyltryptamine ( 5-MeODMT ) decreased the Q9UBK8 and increased the PSR in spinal rats . 4 . Ketanserin antagonized the effects of 5-MeODMT without antagonizing the effects of 8-OH-DPAT . 5 . Cinanserin had similar effects to those of ketanserin . 6 . These results suggest that both P08908 and 5-HT2 receptors mediate Q9UBK8 inhibition and PSR augmentation in the spinal reflexes of spinal rats , and that the 5-HT2 receptor has a supraspinal tonic excitatory influence on the PSR in intact rats . Evidence that the metabotropic glutamate receptor 5 antagonist MPEP may act as an inhibitor of the norepinephrine transporter in vitro and in vivo . The mechanisms through which blockade of metabotropic glutamate receptors 5 ( P41594 ) results in anxiolytic and antidepressant effects are currently unknown . In the present study , we therefore hypothesized that the anxiolytic- and antidepressant-like profile of the noncompetitive P41594 receptor antagonist 2-ethyl-6-(phenylethynyl)-pyridine ( MPEP ) may be mediated by inhibition of the norepinephrine transporter ( NET ) . Accordingly , we first examined the potency of MPEP to bind to or inhibit uptake at the NET as well as the dopamine and serotonin transporters ( Q01959 and P31645 , respectively ) . We also examined the simultaneous in vivo effects of MPEP and desipramine ( DB01151 ) on both NE-like oxidation current in the amygdala ( Q9BXS0 ) and cell firing in the locus coeruleus ( LC ) by means of differential pulse voltammetry ( DPV ) coupled with electrophysiology . MPEP completely displaced the binding of [ 3H ] -nisoxetine on human NET with a pKi of 6.63 +/- 0.02 . In addition , MPEP was able to inhibit [ 3H ] -NE uptake in LLCPK cells expressing human NET , with a pIC50 of 5.55 +/- 0.09 . In vivo DPV data revealed that both MPEP ( 30 mg/kg i.p. ) and DB01151 ( 10 mg/kg i.p. ) significantly increased NE-like voltammetric responses levels in the Q9BXS0 , whereas both compounds also significantly decreased cell firing monitored concomitantly from the second microelectrode in the LC . Collectively , the results of the present study provide potential new mechanisms through which MPEP exerts its anxiolytic and antidepressant effects . 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 . Role of leptin on the expression of low density lipoprotein receptor . BACKGROUND & OBJECTIVES : Leptin resistance oriented hyperleptinaemia is a common problem in obese subjects in association with hypercholesterolaemia . The most common target for hypercholesterolaemia is impaired low density lipoprotein receptor ( P01130 ) . This study was carried out to investigate whether any alteration in P01130 expression could explain the occurrence of hypercholesterolaemia in the event of hyperleptinaemia . METHODS : Expression of P01130 and Q12772 ( sterol regulatory element binding protein 2 ) were examined in HepG2 cells by RT-PCR and Western blotting . O60674 inhibitor II was used to verify the effect of JAK- P35610 ( Janus Kinase-Signal Transducer and Activator of Transcription ) pathway ( common mediator for cytokine signaling ) . Co-localization of P01130 and insulin receptor ( IR ) was examined by confocal microscopy . RESULTS : Leptin was found to reduce the expression of P01130 and its transcription factor Q12772 . On the other hand , a weak signal for stimulation of P01130 by leptin was noted to be mediated by O60674 pathway . But the joint effect of the two signaling pathways kept P01130 only in depressed mode in presence of leptin . Confocal microscopy showed that P01130 made an intensively co-localized complex with insulin receptor in presence of leptin . INTERPRETATION & CONCLUSIONS : Our results show that though leptin stimulates P01130 expression very weakly through JAK- P35610 signaling pathway , it mainly imposes inhibition on P01130 expression by inhibiting transcription factor Q12772 . The inter-association between P01130 and IR may be a reason to render P01130 functionally inactive in presence of leptin . 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 . 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 . Characterization of DB02527 degradation by phosphodiesterases in the accessory olfactory system . To characterize the potential role of DB02527 in pheromone transduction , we have examined the occurrence of cyclic nucleotide phosphodiesterases ( PDEs ) in the mouse vomeronasal organ ( VNO ) . We show that the DB02527 -specific isoforms P27815 and Q08499 are found preferentially in the apical and basal layers , respectively , of the VNO neuroepithelium and in the rostral ( P27815 ) and caudal ( Q08499 ) portions of the accessory olfactory bulb glomerular layer . Assays for DB02527 hydrolysis showed that PDE activity in VNO homogenates was about half that measured in the cerebral cortex and olfactory epithelium , and the proportion of total activity inhibited by rolipram , a DB05876 -specific inhibitor , was approximately 40 % . Activity in the VNO was enhanced 60 % by Ca(2+) and calmodulin ( P62158 ) , implicating the presence of Ca(2+)/ P62158 -dependent PDE1 . Zaprinast , which is known to inhibit Q14123 isoforms , completely suppressed Ca(2+)/ P62158 -stimulated activity and , together , zaprinast and rolipram inhibited DB02527 hydrolysis by approximately 70 % . Our results suggest that PDE1 and DB05876 isoforms are the primary source of DB02527 degradation in the VNO . Decreased expression and activity of DB02527 phosphodiesterases in cardiac hypertrophy and its impact on beta-adrenergic DB02527 signals . RATIONALE : Multiple cyclic nucleotide phosphodiesterases ( PDEs ) degrade DB02527 in cardiomyocytes but the role of PDEs in controlling DB02527 signaling during pathological cardiac hypertrophy is poorly defined . OBJECTIVE : Evaluate the beta-adrenergic regulation of cardiac contractility and characterize the changes in cardiomyocyte DB02527 signals and DB02527 -PDE expression and activity following cardiac hypertrophy . METHODS AND RESULTS : Cardiac hypertrophy was induced in rats by thoracic aortic banding over a time period of 5 weeks and was confirmed by anatomic measurements and echocardiography . Ex vivo myocardial function was evaluated in Langendorff-perfused hearts . Engineered cyclic nucleotide-gated ( CNG ) channels were expressed in single cardiomyocytes to monitor subsarcolemmal DB02527 using whole-cell patch-clamp recordings of the associated CNG current ( I(CNG) ) . PDE variant activity and protein level were determined in purified cardiomyocytes . Aortic stenosis rats exhibited a 67 % increase in heart weight compared to sham-operated animals . The inotropic response to maximal beta-adrenergic stimulation was reduced by approximately 54 % in isolated hypertrophied hearts , along with a approximately 32 % decrease in subsarcolemmal DB02527 levels in hypertrophied myocytes . Total DB02527 hydrolytic activity as well as PDE3 and DB05876 activities were reduced in hypertrophied myocytes , because of a reduction of Q14432 , P27815 , and Q07343 , whereas Q08499 was unchanged . Regulation of beta-adrenergic DB02527 signals by PDEs was blunted in hypertrophied myocytes , as demonstrated by the diminished effects of DB07954 ( 100 micromol/L ) and of both the PDE3 inhibitor cilostamide ( 1 micromol/L ) and the DB05876 inhibitor Ro 201724 ( 10 micromol/L ) . CONCLUSIONS : Beta-adrenergic desensitization is accompanied by a reduction in DB02527 -PDE and an altered modulation of beta-adrenergic DB02527 signals in cardiac hypertrophy . Gene expression and protein localization of calmodulin-dependent phosphodiesterase during ontogenesis of chick retina . P62158 -dependent phosphodiesterase ( PDE1 ) is a key enzyme in cyclic nucleotides metabolism . We studied its gene expression and protein localization during retinal development in chick embryos . Western blot and densitometric analysis demonstrated that the expression of the three isoforms changed during development . P54750 was highly expressed at the early stages and decreased as development proceeded . Q01064 expression remained relatively low and constant over time . Q14123 showed a prominent increase ( 13-fold ) between embryonic day ( E ) 7 and E13 , followed by a moderate increase between E13 and postnatal day ( P ) 1 . The presence of the enzyme in the different retinal locations was strongly modulated by development . P54750 immunostaining was first detected at the ganglion cell level ( E7 ) , then in the outer retina ( E15-E21 ) . At Q15084 , the immunostaining was confined in the optic fiber layer . Isoform C immunolocalization followed the same inner-outer pattern as isoform A . At 5 days posthatching ( Q15084 ) , the immunoreactivity was restricted , as well as for the isoform A , in the optic fiber layer . The isoform B immunolabelling was low and evenly distributed across the retina at all stages . The different developmental profiles of P54750 , Q01064 , and Q14123 induced a temporal modulation in cyclic nucleotides concentration , suggesting specific roles of this enzyme in the morphofunctional development of retinal circuitry . Activation of 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase during high fat diet feeding . The liver plays a central role in regulating cholesterol homeostasis . High fat diets have been shown to induce obesity and hyperlipidemia . Despite considerable advances in our understanding of cholesterol metabolism , the regulation of liver cholesterol biosynthesis in response to high fat diet feeding has not been fully addressed . The aim of the present study was to investigate mechanisms by which a high fat diet caused activation of liver 3-hydroxy-3-methylglutaryl coenzyme A reductase ( P04035 ) leading to increased cholesterol biosynthesis . Mice were fed a high fat diet ( 60 % kcal fat ) for 5weeks . High fat diet feeding induced weight gain and elevated lipid levels ( total cholesterol and triglyceride ) in both the liver and serum . Despite cholesterol accumulation in the liver , there was a significant increase in hepatic P04035 mRNA and protein expression as well as enzyme activity . The DNA binding activity of sterol regulatory element binding protein ( SREBP ) -2 and specific protein 1 ( Sp1 ) were also increased in the liver of mice fed a high fat diet . To validate the in vivo findings , HepG2 cells were treated with palmitic acid . Such a treatment activated Q12772 as well as increased the mRNA and enzyme activity of P04035 leading to intracellular cholesterol accumulation . Inhibition of Sp1 by siRNA transfection abolished palmitic acid-induced Q12772 and P04035 mRNA expression . These results suggest that Sp1-mediated Q12772 activation contributes to high fat diet induced P04035 activation and increased cholesterol biosynthesis . This may play a role in liver cholesterol accumulation and hypercholesterolemia . 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 .

MH_train_1546

MH_train_1546

MH_train_1546

interacts_with DB00501?

multiple_choice

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 . Tandutinib , an oral , small-molecule inhibitor of P36888 for the treatment of AML and other cancer indications . An improved understanding of acute myelogenous leukemia ( AML ) over the past two decades has led to a characterization of associated recurring cytogenetic abnormalities . AML is often driven by the overexpression or constitutive activation of receptor tyrosine kinases such as P36888 ( P36888 ) , which serves as a good therapeutic target . Millennium Pharmaceuticals Inc 's DB05465 ( DB05465 ) is an orally active inhibitor of P36888 kinase and family members P09619 beta and c-Kit . Tandutinib inhibited P36888 phosphorylation , downstream signaling and malignant growth in vitro and in animal models . The drug exhibited limited activity as a single agent in phase I and II clinical trials in patients with AML and myelodysplastic syndrome , but displayed promising antileukemic activity ( 90 % complete remissions ) in a phase I/II trial in patients with newly diagnosed AML when administered in combination with cytarabine and daunorubicin . Phase II clinical trials for DB05465 are ongoing in patients with AML or renal cell carcinoma . Mode of action of agomelatine : synergy between melatonergic and P28335 receptors . OBJECTIVES : The association between depression and circadian rhythm disturbances is well established and successful treatment of depressed patients is accompanied by restoration of circadian rhythms . The new antidepressant agomelatine is an agonist of melatonergic MT₁/MT₂ receptors as well as an antagonist of serotonergic P28335 receptors . Animal studies showed that agomelatine resynchronizes disturbed circadian rhythms and reduces depression-like behaviour . METHODS : This review analyzes results from different experimental studies . RESULTS : Recent data on the effects of agomelatine on cellular processes involved in antidepressant mechanisms have shown that the drug is able to increase the expression of brain-derived neurotrophic factor in prefrontal cortex and hippocampus , as well as the expression of activity-regulated cytoskeleton associated protein ( Arc ) in the prefrontal cortex . In line with this , prolonged treatment with agomelatine increases neurogenesis within the hippocampus , particularly via enhancement of neuronal cell survival . DB06594 attenuates stress-induced glutamate release in the prefrontal/frontal cortex . Treatment with P28335 antagonists or melatonin alone failed to reproduce these effects . CONCLUSIONS : The unique mode of action of agomelatine may improve the management of major depression by counteracting the pathogenesis of depression at cellular level , thereby relieving the symptoms of depression . These effects are suggested to be due to a synergistic action on MT₁/MT₂ and P28335 receptors . The alarmin P09429 acts in synergy with endogenous and exogenous danger signals to promote inflammation . The nuclear protein P09429 has previously been demonstrated to act as an alarmin and to promote inflammation upon extracellular release , yet its mode of action is still not well defined . Access to highly purified P09429 preparations from prokaryotic and eukaryotic sources enabled studies of activation of human PBMC or synovial fibroblast cultures in response to P09429 alone or after binding to cofactors . P09429 on its own could not induce detectable P05231 production . However , strong enhancing effects on induction of proinflammatory cytokine production occurred when the protein associated with each of the separate proinflammatory molecules , rhIL-1beta , the O00206 ligand LPS , the Q9NR96 ligand CpG-ODN , or the Q15399 - O60603 ligand Pam3CSK4 . The bioactivities were recorded in cocultures with preformed P09429 complexes but not after sequential or simultaneous addition of P09429 and the individual ligands . Individual A-box and B-box domains of P09429 had the ability to bind LPS and enhance P05231 production . Heat denaturation of P09429 eliminated this enhancement . Cocultures with P09429 and other proinflammatory molecules such as P01375 , O14788 , or Q14116 did not induce enhancement . P09429 thus acts broadly with many but not all immunostimulatory molecules to amplify their activity in a synergistic manner . P03372 -alpha signaling in growth of the ventral prostate : comparison of neonatal growth and postcastration regrowth . A role for estrogen receptor ( ER ) -alpha in branching morphogenesis in the ventral prostate ( VP ) has previously been demonstrated ; in the VP of ERalpha(-/-) mice , there are fewer side branches than in wild-type littermates . In the present study , we show that in the postnatal VP , fibroblast growth factor 10 ( O15520 ) is expressed in wild-type mice but not in ERalpha(-/-) mice , and because branching involves proliferation pathways also used in malignant growth , we investigated whether branching during regrowth of the VP after castration involves ERalpha and O15520 . ERalpha was not detectable in the prostates of sham-operated or castrated mice but was expressed in the prostatic epithelium between d 3 and 5 after testosterone replacement . Blocking either ERalpha or ERbeta with DB00947 had no detectable effects on epithelial cell proliferation during regrowth by testosterone . The ERalpha agonist , propylpyrazoletriol , did not induce regrowth by itself , but exposure to propylpyrazoletriol on d 3-5 of testosterone replacement resulted in cyclin D1-positive cells in the ductal epithelium , invasion of O15520 -positive immune cells in the regrowing prostate , and budding 14 d later . DB00624 replacement alone did not induce cyclin D1 , O15520 , or bud formation . These results indicate that stimulation of ERalpha is essential for ductal branching during postnatal prostate growth . During regrowth after castration , there is a window in time when selective stimulation of ERalpha can also induce ductal branching . The O15520 for this growth comes from the immune system , not from the prostatic mesenchyme . 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 . DCEBIO stimulates Cl- secretion in the mouse jejunum . We investigated the effects of 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one(DCEBIO) on the Cl- secretory response of the mouse jejunum using the Ussing short-circuit current ( Isc ) technique . DCEBIO stimulated a concentration-dependent , sustained increase in Isc ( EC50 41 +/- 1 microM ) . Pretreating tissues with 0.25 microM forskolin reduced the concentration-dependent increase in Isc by DCEBIO and increased the EC50 ( 53 +/- 5 microM ) . DB00887 blocked ( 82 +/- 5 % ) the DCEBIO-stimulated Isc consistent with Cl- secretion . DCEBIO was a more potent stimulator of Cl- secretion than its parent molecule , 1-ethyl-2-benzimidazolinone . DB01016 or P16860 reduced the DCEBIO-stimulated Isc by > 80 % indicating the participation of P13569 in the DCEBIO-stimulated Isc response . Clotrimazole reduced DCEBIO-stimulated Isc by 67 +/- 15 % , suggesting the participation of the intermediate conductance Ca2+-activated K+ channel ( IKCa ) in the DCEBIO-activated Isc response . In the presence of maximum forskolin ( 10 microM ) , the DCEBIO response was reduced and biphasic , reaching a peak response of the change in Isc of 43 +/- 5 microA/cm2 and then falling to a steady-state response of 17 +/- 10 microA/cm2 compared with DCEBIO control tissues ( 61 +/- 6 microA/cm2 ) . The forskolin-stimulated Isc in the presence of DCEBIO was reduced compared with forskolin control tissues . Similar results were observed with DCEBIO and 8-BrcAMP where adenylate cyclase was bypassed . H89 , a PKA inhibitor , reduced the DCEBIO-activated Isc , providing evidence that DCEBIO increased Cl- secretion via a DB02527 /PKA-dependent manner . These data suggest that DCEBIO stimulates Cl- secretion of the mouse jejunum and that DCEBIO targets components of the Cl- secretory mechanism . 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 . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . 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 . 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 . 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 . Genetic dissection of atypical antipsychotic-induced weight gain : novel preliminary data on the pharmacogenetic puzzle . Atypical antipsychotics such as clozapine represent a significant improvement over typical antipsychotics in the treatment of schizophrenia , particularly regarding extrapyramidal symptoms . Despite their benefits , use is limited by the occurrence of adverse reactions such as sedation and weight gain . This article provides a comprehensive review and discussion of obesity-related pathways and integrates these with the known mechanisms of atypical antipsychotic action to identify candidate molecules that may be disrupted during antipsychotic treatment . Novel preliminary data are presented to genetically dissect these obesity pathways and elucidate the genetic contribution of these candidate molecules to clozapine-induced weight gain . There is considerable variability among individuals with respect to the ability of clozapine to induce weight gain . Genetic predisposition to clozapine-induced weight gain has been suggested . Therefore , genetic variation in these candidate molecules may predict patient susceptibility to clozapine-induced weight gain . This hypothesis was tested for 10 genetic polymorphisms across 9 candidate genes , including the serotonin 2C , 2A , and 1A receptor genes ( P28335 /2A/1A ) ; the histamine H1 and H2 receptor genes ( P35367 / P25021 ) ; the cytochrome P450 1A2 gene ( P05177 ) ; the beta3 and alpha,alpha-adrenergic receptor genes ( P13945 /ADRAIA ) ; and tumor necrosis factor alpha ( P01375 ) . Prospective weight gain data were obtained for 80 patients with schizophrenia who completed a structured clozapine trial . Trends were observed for P13945 , ADRA1A , P01375 , and P28335 ; however , replication in larger , independent samples is required . Although in its infancy , psychiatric pharmacogenetics will in the future aid clinical practice in the prediction of response and side effects , such as antipsychotic-induced weight gain , and minimize the current " trial and error " approach to prescribing . 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 . 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 . Different transport properties between famotidine and cimetidine by human renal organic ion transporters ( SLC22A ) . P25021 antagonist famotidine and cimetidine are commonly used for treatment of gastrointestinal ulcer diseases . Inasmuch as these drugs are mainly secreted by renal tubules , dosages have been adjusted according to renal function . Although many studies have been performed on the molecular mechanisms of renal handling of cimetidine , little is known about that of famotidine . In this study , to examine the recognition and transport of famotidine by human organic anion transporters ( OATs ; Q4U2R8 , Q8TCC7 ) and human organic cation transporter ( O75051 ; O15244 ) , the uptake studies using Xenopus laevis oocytes were performed in comparison with cimetidine . The half-maximal inhibitory concentrations of famotidine for [3H]estrone sulfate transport by Q8TCC7 and [14C]tetraethylammonium transport by O15244 ( 300 microM and 1.8 mM , respectively ) were higher than those of cimetidine ( 53 and 67 microM , respectively ) . While cimetidine inhibited p-[14C]aminohippurate transport by Q4U2R8 in a concentration dependent manner , famotidine did not affect it at 5 mM . In addition , Q8TCC7 mediated famotidine uptake , but Q4U2R8 and O15244 did not show famotidine transport . These results indicate that there are marked differences between famotidine and cimetidine in the recognition and transport by organic ion transporters and that Q8TCC7 contributes to the renal tubular secretion of famotidine . Present findings should be useful information to understand the renal handling of famotidine and cimetidine . Molecular characterization of gallbladder cancer using somatic mutation profiling . Gallbladder cancer is relatively uncommon , with a high incidence in certain geographic locations , including Latin America , East and South Asia , and Eastern Europe . Molecular characterization of this disease has been limited , and targeted therapy options for advanced disease remain an open area of investigation . In the present study , surgical pathology obtained from resected gallbladder cancer cases ( n = 72 ) was examined for the presence of targetable , somatic mutations . All cases were formalin fixed and paraffin embedded ( FFPE ) . Two approaches were used : ( a ) mass spectroscopy-based profiling for 159 point ( " hot spot " ) mutations in 33 genes commonly involved in solid tumors and ( b ) next-generation sequencing ( NGS ) platform that examined the complete coding sequence of in 182 cancer-related genes . Fifty-seven cases were analyzed for hot spot mutations ; and 15 , for NGS . Fourteen hot spot mutations were identified in 9 cases . Of these , P01116 mutation was significantly associated with poor survival on multivariate analysis . Other targetable mutations included P42336 ( n = 2 ) and Q9UM73 ( n = 1 ) . On NGS , 26 mutations were noted in 15 cases . P04637 and P19957 kinase pathway ( Q15831 , Q6R327 , P49815 ) mutations were common . One case had O15520 amplification , whereas another had P11487 -TACC gene fusion , not previously described in gallbladder cancer . In conclusion , somatic mutation profiling using archival FFPE samples from gallbladder cancer is feasible . NGS , in particular , may be a useful platform for identifying novel mutations for targeted therapy . 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 . Receptor tyrosine kinase pathway analysis sheds light on similarities between clear-cell sarcoma and metastatic melanoma . To highlight possible similarities and differences in receptor tyrosine kinase ( RTK ) and downstream signalling activation profiles between clear-cell sarcomas ( O14618 ) and metastatic melanomas ( MM ) , frozen , and paired-matched fixed samples of six O14618 with Q01844 rearrangement ( Q01844 + ) , five O14618 without Q01844 rearrangement ( Q01844 - ) , and seven MM were investigated by means of biochemical , immunohistochemical , Q5TCZ1 , molecular analyses , and immunofluorescence confocal microscopy . Fixed samples of a further 10 O14618 and 14 MM were investigated by means of sequencing for P15056 , P01111 , and P01116 mutations and Q5TCZ1 analyses for the gain of chromosomes 22 and 8 . RTK analysis of all O14618 /MM samples showed activation of short-form ( sf ) recepteur d'origine nantais ( Q04912 ) RTK and of P09619 , MET , and P21860 . Analysis of downstream signaling revealed consistent phosphorylation patterns of PI3K/AKT , RSK , and the P42345 targets S6 and 4EBP1 . Analysis of frozen and fixed material from 21 O14618 and 21 MM showed the presence of the V600E P15056 mutation in 2/12 Q01844 + and 3/9 Q01844 - O14618 and 9/21 MM and demonstrated a significant ( P < 0.001 ) correlation between the gain of chromosomes 22 and 8 and Q01844 - O14618 . Our results show that P15056 mutation can also be present in O14618 and support the proposed aberration of chromosomes 22 and 8 as a possibly useful nonrandom hallmark of Q01844 - O14618 . Besides , they broaden the spectrum of the similarities of RTK pathway activation between O14618 and MM , thus suggesting that new drugs found to be active in melanoma and Q04912 inhibitors could have a role in O14618 treatment . © 2011 Wiley Periodicals , Inc . 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 . Genomic landscape of non-small cell lung cancer in smokers and never-smokers . We report the results of whole-genome and transcriptome sequencing of tumor and adjacent normal tissue samples from 17 patients with non-small cell lung carcinoma ( NSCLC ) . We identified 3,726 point mutations and more than 90 indels in the coding sequence , with an average mutation frequency more than 10-fold higher in smokers than in never-smokers . Novel alterations in genes involved in chromatin modification and DNA repair pathways were identified , along with Q9UI36 , P13569 , P78509 , Q2M3G0 , and P14210 . Deep digital sequencing revealed diverse clonality patterns in both never-smokers and smokers . All validated EFGR and P01116 mutations were present in the founder clones , suggesting possible roles in cancer initiation . Analysis revealed 14 fusions , including P08922 and Q9UM73 , as well as novel metabolic enzymes . Cell-cycle and JAK- P35610 pathways are significantly altered in lung cancer , along with perturbations in 54 genes that are potentially targetable with currently available drugs .

MH_train_1547

MH_train_1547

MH_train_1547

interacts_with DB08820?

multiple_choice

P15941 -C oncoprotein promotes P36888 receptor activation in acute myeloid leukemia cells . Blasts from approximately one-third of patients with acute myeloid leukemia ( AML ) harbor activating mutations in the P07333 -like tyrosine kinase 3 ( P36888 ) receptor tyrosine kinase that confer a poor prognosis . The Mucin 1-C-terminal subunit ( P15941 -C ) oncoprotein is aberrantly expressed in AML blasts and stem cells ; however , there is no known interaction between P15941 -C and P36888 . The present studies demonstrate that P15941 -C associates with wild-type and mutant P36888 in AML cells . Targeting P15941 -C with the cell-penetrating peptide inhibitor GO-203 disrupts P15941 -C/ P36888 complexes and downregulates P36888 activation . GO-203 treatment of AML cells was also associated with inhibition of the P36888 downstream effectors AKT , extracellular signal-regulated kinase , and P42229 . The results further show that AML cells with P36888 -activating mutations and resistant to the P36888 inhibitor midostaurin/PKC412 are sensitive to GO-203-induced growth arrest and death . Moreover , GO-203 increases sensitivity of mutant P36888 AML cells to P36888 inhibitor treatment . These results indicate that P15941 -C contributes to P36888 activation in AML cells and that targeting P15941 -C inhibits the P36888 signaling pathway . Our findings support the development of P15941 -C inhibitors alone and in combination with agents that target P36888 for the treatment of wild-type and mutant P36888 AML . DB01169 induces cardiac fibroblast apoptosis in vitro and in vivo by up-regulating TGF-β1 expression . DB01169 ( As2O3 ; ATO ) is clinically effective in treating acute promyelocytic leukemia ( APL ) ; however , it frequently causes cardiotoxic effects . This study was designed to investigate whether ATO could induce apoptosis of cardiac fibroblasts ( CFs ) that play very important roles in maintaining the structure integrity and function of the heart . Cardiac fibroblasts from guinea pigs administered with ATO ( 1mg/kgbw ) were used to test the pro-apoptotic role of ATO in vivo . The current study demonstrated that ATO induced morphological characteristics of apoptosis and P42574 activation in CFs of guinea pigs along with a significant up-regulation in TGF-β1 protein expression , Bax/Bcl-2 ratio and P27361 /2 phosphorylation . In vitro MTT assay showed that ATO remarkably reduced the viability of cultured cardiac fibroblasts ( NRCFs ) from neonatal rat in a concentration- and time-dependent manner . Consistent with the notions in vivo , ATO significantly induced the apoptosis in NRCFs , dramatically up-regulated TGF-β1 protein level and Bax/Bcl-2 ratio in a time-dependent fashion and activated P42574 and P27361 /2 . Finally , pretreatment with LY364947 , an inhibitor of TGF-β signaling could apparently reverse these changes . We therefore conclude that TGF-β is functionally linked to P27361 /2 and that TGF-β signaling is responsible for ATO-induced CFs apoptosis , which provides a novel mechanism of ATO related cardiac toxicology . Multiple endocytic signals in the C-terminal tail of the cystic fibrosis transmembrane conductance regulator . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -dependent protein kinase ( PKA ) -activated chloride channel that is localized to the plasma membrane and endosomal compartment . Endosomal targeting of P13569 is attributed to the DB00135 (1424)-based internalization signal , identified in the C-terminal tail of the channel . Mutation of the DB00135 (1424) residue could partly inhibit the endocytosis of P13569 and its association with the adapter protein P05549 . To reveal additional endosomal targeting signals , site-directed mutagenesis of both a chimaera , composed of a truncated form of interleukin 2 receptor alpha chain ( TacT ) and the C-terminal tail of P13569 ( Ct ) , and the full-length P13569 was performed . Morphological and functional assays revealed the presence of multiple internalization motifs at the C-terminus , consisting of a phenylalanine-based motif ( DB00120 (1413) ) and a bipartite endocytic signal , comprising a tyrosine ( DB00135 (1424) ) and a di- DB00149 -based ( DB00149 (1430)- DB00149 ) motif . Whereas the replacement of any one of the three internalization motifs with alanine prevented the endocytosis of the TacT-Ct chimaera , mutagenesis of DB00120 (1413)- DB00149 impaired the biosynthetic processing of P13569 , indicating that DB00120 (1413) is indispensable for the native structure of P13569 . In contrast , replacement of DB00149 (1430)- DB00149 - and DB00135 (1424)-based signals with alanine increased the cell-surface density of both the chimaeras and P13569 in an additive manner . These results suggest that the internalization of P13569 is regulated by multiple endocytic sorting signals . 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 . Oxidative stress induces extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase in cystic fibrosis lung epithelial cells : Potential mechanism for excessive P10145 expression . Cystic fibrosis ( CF ) is a lethal disease caused by defective function of the cftr gene product , the CF transmembrane conductance regulator ( P13569 ) that leads to oxidative damage and excessive inflammatory response in lungs of CF patients . We here report the effects of oxidative stress ( hyperoxia , 95 % O(2) ) on the expression of pro-inflammatory interleukin ( IL ) -8 and P25024 /2 receptors in two human CF lung epithelial cell lines ( IB3-1 , with the heterozygous F508del/W1282X mutation and CFBE41o- with the homozygous F508del/F508del mutation ) and two control non-CF lung epithelial cell lines ( S9 cell line derived from IB3-1 after correction with wtCFTR and the normal bronchial cell line 16HBE14o- ) . Under oxidative stress , the expression of P10145 and P25024 /2 receptors was increased in CF , corrected and normal lung cell lines . The effects of oxidative stress were also investigated by measuring the transcription nuclear factor kappaB ( NF-kappaB ) and activator protein-1 ( AP-1 ) activities . Under oxidative stress , no increase of NF-kappaB activation was observed in CF lung cells in contrast to that observed in normal and corrected CF lung cells . The signalling of mitogen-activated protein ( Q96HU1 ) kinases was further studied . We demonstrated that extracellular signal-regulated kinase ( P27361 /2 ) and AP-1 activity was markedly enhanced in CF but not non-CF lung cells under oxidative stress . Consistently , inhibition of P27361 /2 in oxidative stress-exposed CF lung cells strongly decreased both the P10145 production and P25024 /2 expression . Therefore , targeting of P27361 /2 Q96HU1 kinase may be critical to reduce oxidative stress-mediated inflammation in lungs of CF patients . 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 . 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 . O60760 conjugation of organophosphorus pesticides yields S-phospho- , S-aryl- , and S-alkylglutathione derivatives . Pesticide detoxification is a central feature of selective toxicity and safety evaluation . Two of the principal enzymes involved are DB00143 S-transferases ( GSTs ) and cytochrome P450s acting alone and together . More than 100 pesticides are organophosphorus ( OP ) compounds , but with few exceptions , their DB00143 conjugates have not been directly observed in vitro or in vivo . The major insecticides chlorpyrifos ( CP ) and diazinon are of particular interest as multifunctional substrates with diverse metabolites , while ClP(S)(OEt) 2 and the cotton defoliant tribufos are possible precursors of phosphorylated DB00143 conjugates . Formation of DB00143 conjugates by Q86UG4 with DB00143 was studied in vitro with and without metabolic activation by human liver microsomes or P450 3A4 with NADPH . Metabolites were analyzed by liquid chromatography-electrospray ionization-mass spectrometry ( LC- P19957 -MS ) . Five DB00143 conjugates were identified from CP and chlorpyrifos oxon ( CPO ) , i.e. , GSCP and GSCPO in which the 6-chloro substituent of CP and CPO , respectively , is displaced by DB00143 ; S-(3,5,6-trichloropyridin-2-yl)glutathione ; S-(3,5-dichloro-6-hydroxypyridin-2-yl)glutathione ; and S-ethylglutathione . Q86UG4 of a human liver microsomal preparation but not P450 3A4 with DB00143 metabolized CP to GSCP . With Q86UG4 and DB00143 , diazinon and diazoxon gave S-(2-isopropyl-4-methylpyrimidin-6-yl)glutathione and ClP(S)(OEt) 2 yielded P63092 (S)(OEt) 2 . With microsomes , NADPH , Q86UG4 , and DB00143 tribufos gave P63092 (O)(SBu) 2 . The liver of intraperitoneally treated mice contained GSCP from CP , P63092 (S)(OEt) 2 from ClP(S)(OEt) 2 , and P63092 (O)(SBu) 2 from tribufos . P63092 (S)(OEt) 2 and P63092 (O)(SBu) 2 are the first S-phosphoglutathione metabolites observed in vitro and in vivo directly by LC- P19957 -MS . Nine other OP pesticides gave only O-dealkylation in the Q86UG4 / DB00143 system . Q86UG4 -catalyzed metabolism joins P450s and hydrolases as important contributors to OP detoxification . DB00227 prevents angiotensin II-induced cardiac hypertrophy in cultured neonatal rat heart cells . Angiotensin II activates P01112 , and mediates cardiac hypertrophic growth through the type 1 angiotensin II receptor in cardiac myocytes . An inhibitor of 3-hydroxy-3-methyglutaryl-coenzyme A ( HMG- DB01992 ) reductase has been shown to block the post-translational farnesylation of P01112 and inhibit protein synthesis in several cell types . Primary cultures of neonatal cardiac myocytes were used to determine whether P04035 inhibitors , lovastatin , simvastatin and pravastatin inhibit the angiotensin II-induced hypertrophic growth . Angiotensin II ( 10(-6) M ) significantly increased protein-DNA ratio , RNA-DNA ratio , ratios of protein synthesis and mitogen-activated protein ( Q96HU1 ) kinase activity . Lipid-soluble P04035 inhibitors , lovastatin ( 10(-6) M ) and simvastatin ( 10(-6) M ) partially and significantly inhibited the angiotensin II-induced increases in these parameters , but a water-soluble P04035 inhibitor , pravastatin ( 10(-6) M ) did not . Mevalonate ( 10(-4) M ) overcame the inhibitory effects of lovastatin and simvastatin on angiotensin II-induced increases in these parameters . A selective protein kinase C inhibitor , calphostin C ( 10(-6) M ) partially and significantly prevented angiotensin II-induced increases in these parameters , and treatment with both lovastatin and calphostin C inhibited completely . Angiotensin II increased P01112 activity and membrane association , and lovastatin inhibited them . These studies demonstrate that a lipid-soluble P04035 inhibitor , lovastatin , may prevent angiotensin II-induced cardiac hypertrophy , at least in part , through P01112 / Q96HU1 kinase pathway , which is linked to mevalonate metabolism . An immuno-chemo-proteomics method for drug target deconvolution . Chemical proteomics is an emerging technique for drug target deconvolution and profiling the toxicity of known drugs . With the use of this technique , the specificity of a small molecule inhibitor toward its potential targets can be characterized and information thus obtained can be used in optimizing lead compounds . Most commonly , small molecules are immobilized on solid supports and used as affinity chromatography resins to bind targets . However , it is difficult to evaluate the effect of immobilization on the affinity of the compounds to their targets . Here , we describe the development and application of a soluble probe where a small molecule was coupled with a peptide epitope which was used to affinity isolate binding proteins from cell lysate . The soluble probe allowed direct verification that the compound after coupling with peptide epitope retained its binding characteristics . The P17252 inhibitor Bisindolylmaleimide-III was coupled with a peptide containing the FLAG epitope . Following incubation with cellular lysates , the compound and associated proteins were affinity isolated using anti-FLAG antibody beads . Using this approach , we identified the known Bisindolylmaleimide-III targets , P17252 , GSK3-beta , CaMKII , adenosine kinase , P24941 , and quinine reductase type 2 , as well as previously unidentified targets PKAC-alpha , prohibitin , P21796 and heme binding proteins . This method was directly compared to the solid-phase method ( small molecule was immobilized to a solid support ) providing an orthogonal strategy to aid in target deconvolution and help to eliminate false positives originating from nonspecific binding of the proteins to the matrix . 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 . Coadministrating luteolin minimizes the side effects of the aromatase inhibitor letrozole . P11511 inhibitors ( AIs ) have been used as adjuvant therapeutic agents for breast cancer . Their adverse side effect on blood lipid is well documented . Some natural compounds have been shown to be potential AIs . In the present study , we compared the efficacy of the flavonoid luteolin to the clinically approved AI letrozole ( DB01006 ; Novartis Pharmaceuticals , East Hanover , NJ ) in a cell and a mouse model . In the in vitro experimental results for aromatase inhibition , the Ki values of luteolin and letrozole were estimated to be 2.44 µM and 0.41 nM , respectively . Both letrozole and luteolin appeared to be competitive inhibitors . Subsequently , an animal model was used for the comparison . P11511 -expressing MCF-7 cells were transplanted into ovariectomized athymic mice . Luteolin was given by mouth at 5 , 20 , and 50 mg/kg , whereas letrozole was administered by intravenous injection . Similar to letrozole , luteolin administration reduced plasma estrogen concentrations and suppressed the xenograft proliferation . The regulation of cell cycle and apoptotic proteins-such as a decrease in the expression of Bcl-xL , cyclin-A/D1/E , P24941 /4 , and increase in that of Bax-was about the same in both treatments . The most significant disparity was on blood lipids . In contrast to letrozole , luteolin increased fasting plasma high-density lipoprotein concentrations and produced a desirable blood lipid profile . These results suggested that the flavonoid could be a coadjuvant therapeutic agent without impairing the action of AIs . Significance of O00141 in the regulation of neuronal function . The present brief review highlights the putative role of the serum- and glucocorticoid-inducible-kinase-1 ( O00141 ) in the regulation of neuronal function . O00141 is genomically upregulated by cell shrinkage and by a variety of hormones including mineralocorticoids and glucocorticoids . The kinase is activated by insulin and growth factors via phosphatidylinositide-3-kinase ( P19957 -kinase ) , phosphoinositide-dependent kinase PDK1 and mammalian target of rapamycin mTORC2 . O00141 upregulates ion channels ( e.g. Q14524 , ENaC , P78348 , Q9NQA5 ,6 , ROMK , Kv1.1-5 , KCNEx/ P51787 -5 , Q13002 , VSOAC , ClC2 , P13569 ) , carriers ( e.g. P48764 , Q13621 , P55017 , NaPiIIb , SMIT , P11166 ,4 , P13866 , NaDC , P43003 -5 , Q99624 , Q15758 , 4F2/ O43561 , PepT2 ) , and the Na(+)/K(+)-ATPase . O00141 regulates enzymes ( e.g. glycogen-synthase-kinase-3 , ubiquitin-ligase Q96PU5 , phosphomannose-mutase-2 ) , and transcription factors ( e.g. forkhead transcription factor Foxo3a , β-catenin , nuclear factor-kappa-B ( NFB ) ) . O00141 participates in the regulation of transport , hormone release , neuroexcitability , inflammation , coagulation , cell proliferation and apoptosis . O00141 contributes to regulation of renal Na(+) retention , renal K(+) elimination , salt appetite , gastric acid secretion , intestinal Na(+)/H(+) exchange and nutrient transport , insulin-dependent salt sensitivity of blood pressure , salt sensitivity of peripheral glucose uptake , cardiac repolarization and memory consolidation . Presumably , O00141 contributes to the regulation of diverse cerebral functions ( e.g. memory consolidation , fear retention ) and the pathophysiology of several cerebral diseases ( e.g. Parkinson 's disease , schizophrenia , depression , Alzheimer 's disease ) . Despite multiple O00141 functions , the phenotype of the O00141 knockout mouse is mild and becomes only apparent under challenging conditions . Rescue of amyotrophic lateral sclerosis phenotype in a mouse model by intravenous AAV9-ADAR2 delivery to motor neurons . Amyotrophic lateral sclerosis ( P35858 ) is the most common adult-onset motor neuron disease , and the lack of effective therapy results in inevitable death within a few years of onset . Failure of P42262 RNA editing resulting from downregulation of the RNA-editing enzyme adenosine deaminase acting on RNA 2 ( ADAR2 ) occurs in the majority of P35858 cases and causes the death of motor neurons via a Ca(2+) -permeable AMPA receptor-mediated mechanism . Here , we explored the possibility of gene therapy for P35858 by upregulating ADAR2 in mouse motor neurons using an adeno-associated virus serotype 9 ( AAV9 ) vector that provides gene delivery to a wide array of central neurons after peripheral administration . A single intravenous injection of AAV9-ADAR2 in conditional ADAR2 knockout mice ( AR2 ) , which comprise a mechanistic mouse model of sporadic P35858 , caused expression of exogenous ADAR2 in the central neurons and effectively prevented progressive motor dysfunction . Notably , AAV9-ADAR2 rescued the motor neurons of AR2 mice from death by normalizing Q13148 expression . This AAV9-mediated ADAR2 gene delivery may therefore enable the development of a gene therapy for P35858 . Biophysical and pharmacological characterization of hypotonically activated chloride currents in cortical astrocytes . Rat cortical astrocytes regulate their cell volume in response to hypotonic challenge . This regulation is believed to depend largely on the release of chloride or organic osmolytes through anion channels . Using whole-cell recordings , we identified weakly outwardly rectifying chloride currents that could be activated in response to hypotonic challenge . These currents exhibited the following permeability sequence upon replacement of chloride in the bathing solution with various anions : I- > NO3- > Cl- > Gluc- > or = MeS- > Ise- . Interestingly , extracellular I- , albeit showing the greatest permeability , blocked the currents with an IC50 of approximately 50 mM . Currents were almost completely inhibited by 123 microM P16860 and partially inhibited by 200 microM niflumic acid or 200 microM DIDS . Additionally , the total number of Cl- ions effluxed through the hypotonically activated channels was markedly similar to the total solute efflux during volume regulation . We therefore propose the hypotonically activated chloride channel as a major contributor to volume regulation of astrocytes . To examine potential candidate chloride channel genes expressed by astrocytes , we employed RT-PCR to demonstrate the presence of transcripts for P51788 , 3 , 4 , 5 , and 7 , as well as for P21796 and P13569 in cultured astrocytes . Moreover , we performed immunostaining with antibodies against each of these channels and showed the strongest expression of P51788 and P51790 , strong expression of P51795 and P21796 , weak expression of P51798 and very weak expression of P51793 and P13569 . Intriguingly , although we found at least seven Cl- channel proteins from three different gene families in astrocytes , none appeared to be active in resting cells . The multitargeted receptor tyrosine kinase inhibitor linifanib ( DB06080 ) induces apoptosis through an Akt and glycogen synthase kinase 3β-dependent pathway . The P07333 -like receptor tyrosine kinase 3 ( P36888 ) plays an important role in controlling differentiation and proliferation of hematopoietic cells . Activating mutations in P36888 occur in patients with acute myeloid leukemia ( AML ; 15 % -35 % ) , resulting in abnormal cell proliferation . Furthermore , both adult and pediatric patients with AML harboring the P36888 internal tandem duplication ( ITD ) mutation have a poor prognosis . Several inhibitors have been developed to target mutant P36888 for the treatment of AML , yet the molecular pathways affected by drug inhibition of the mutated P36888 receptor alone have not been characterized as yet . Linifanib ( DB06080 ) is a multitargeted tyrosine kinase receptor inhibitor that suppresses P36888 signaling . In this article , we show that treatment with linifanib inhibits proliferation and induces apoptosis in ITD mutant cells in vitro and in vivo . We show that treatment with linifanib reduces phosphorylation of Akt and glycogen synthase kinase 3β ( GSK3β ) . In addition , we show that inhibition of GSK3β decreases linifanib-induced apoptosis . This study shows the importance of GSK3 as a potential target for AML therapy , particularly in patients with P36888 ITD mutations . Expression and mutational status of treatment-relevant targets and key oncogenes in 123 malignant salivary gland tumours . BACKGROUND : Malignant tumours of the salivary glands ( MSGT ) are rare and pleomorphic entities . Patients with advanced disease may benefit from targeted therapy ; however , specific targets for optimising and personalising treatments are yet to be identified . DESIGN : Immunohistochemistry for C- P10721 , P00533 , P04626 , P15941 , phospho- P42345 , androgen/estrogens/progesterone receptors and Ki67 was carried out and evaluated in terms of progression-free and overall survival . High throughput molecular screening of key oncogenes was done in 107 patients using routine diagnostic methods and Sequenom technology . RESULTS : Several therapy leads were identified , including high levels of P04626 and androgen receptors in salivary duct carcinomas , C- P10721 in myoepithelial carcinomas and P00533 in mucoepidermoid carcinomas . Recurrent mutations involving downstream elements of the P00533 pathway were found in P01112 , notably in tumours with a myoepithelial component , and in other key oncogenes ( P01116 / P01111 /PI3KCA/ P15056 /MAP2K ) . On the other hand , < 1 % of samples had P00533 or P04626 mutations . CONCLUSION : Several tumour subtypes overexpressed targets of directed therapies suggesting potential therapy leads . Genotyping results suggest activation downstream of P00533 in 18 of the 107 samples that could be associated with low efficacy of P00533 inhibitors . Other molecules , such as PI3K/MEK or P42345 inhibitors , may have anti-tumour activity in this subgroup . The high mutation rate in P01112 highlights a novel key oncogenic event in MSGT . 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 . Genetics of idiopathic disseminated bronchiectasis . Bronchiectasis is an abnormal dilation of bronchi , consequent to the destruction of their walls . It is included in the category of obstructive pulmonary diseases , along with chronic obstructive pulmonary disease ( P48444 ) , asthma , and cystic fibrosis . In approximately 50 % of cases , bronchiectasis is associated with underlying conditions ; in the remainder , known causes are not ascertainable ( idiopathic bronchiectasis ) . A search for genetic determinants of this phenotype , with the cystic fibrosis gene as a candidate , has been performed by three independent groups . The results of this search agreed on the association of bronchiectasis with cystic fibrosis gene mutations and polymorphisms . The cystic fibrosis gene is also associated with bronchiectasis due to rheumatoid arthritis and allergic bronchopulmonary aspergillosis . A few other genes have been investigated in idiopathic bronchiectasis , with negative results . Idiopathic bronchiectasis is , therefore , to be considered as an obstructive multifactorial disorder belonging to the category of cystic fibrosis monosymptomatic diseases ( or P13569 -opathies ) , whose pathogenesis is influenced by environmental factors and other undetermined genes . Computation of standard binding free energies of polar and charged ligands to the glutamate receptor P42262 . Accurate calculation of the binding affinity of small molecules to proteins has the potential to become an important tool in rational drug design . In this study , we use the free energy perturbation ( FEP ) method with restraints to calculate the standard binding free energy of five ligands ( ACPA , AMPA , CNQX , DB03759 , and glutamate ) to the glutamate receptor P42262 , which plays an essential role in synaptic transmission . To deal with the convergence problem in FEP calculations with charged ligands , we use a protocol where the ligand is coupled in the binding site while it is decoupled in bulk solution simultaneously . The contributions from the conformational , rotational , and translational entropies to the standard binding free energy are determined by applying/releasing respective restraints to the ligand in bulk/binding site . We also employ the confine-and-release approach , which helps to resolve convergence problems in FEP calculations . Our results are in good agreement with the experimental values for all five ligands , including the charged ones which are often problematic in FEP calculations . We also analyze the different contributions to the binding free energy of each ligand to P42262 and discuss the nature of these interactions . [ Assessment of the hypothalamic-hypophyseal-adrenal axis in patients with chronic obstructive lung disease. Comparison of inhalant with systemic glucocorticoid therapy ] . OBJECTIVE : The action of inhalation and systemic treatment of chronic obstructive pulmonary disease by suppressing the hypothalamo-hypophyseal-adrenal axis was compared in patients with chronic obstructive pulmonary disease ( P48444 ) . PATIENTS AND METHODS : DB01285 ( DB01285 ) and cortisol concentrations were evaluated after a corticotropin-releasing-hormone ( P06850 ) -test in 50 patients ( aged 43 +/- 14 years ) with chronic obstructive pulmonary disease ( P48444 ) receiving inhalant glucocorticoid treatment ( IGC ) , 61 patients ( aged 54 +/- 11 years ) with P48444 on systemic glucocorticoid treatment ( SGC ) and 50 healthy volunteers ( 32 +/- 4 years ) . RESULTS : All 50 patients on IGC had normal P06850 test results . 30 of 61 patients with SGC had decreased cortisol response ( 12 patients had no and 18 a reduced rise in cortisol ) . DB01285 concentration was lower in patients on IGC than in the control group ( basal DB01285 15.6 pg/ml and 24.5 pg/ml , respectively ; after stimulation 40.3 vs 54.4 pg/ml , respectively ) . But systemic glucocorticoid treatment clearly caused suppression of basal ( 12.1 pg/ml ) and stimulated ( 27.4 pg/ml ) DB01285 levels with correspondingly decreased cortisol levels ( basal : 75.1 and 118.7 ng/ml [ IGC ] , respectively , and after stimulation 128.5 and 225.9 ng/ml ) . CONCLUSIONS : Patients with P48444 on inhalant glucocorticoid treatment have a clearly lower risk of adrenal cortical insufficiency than those on oral glucocorticoid treatment . But some suppression of DB01285 secretion is demonstrable even in the former . Clinical significance of these findings seems unlikely . Development of adrenal cortical insufficiency need not be feared in patients treated with inhalant glucocorticoids . 5-Hydroxytryptamine contributes significantly to a reflex pathway by which the duodenal mucosa protects itself from gastric acid injury . Although duodenal mucosal bicarbonate secretion ( DMBS ) is currently accepted as an important defense mechanism against acid-induced duodenal injury , the mechanism and the regulation of DMBS are largely unknown . 5-HT may regulate DMBS , but little is known about its physiological relevance in DMBS and the underlying mechanism(s) . Thus , the aims of the present study were to demonstrate the role of 5-HT in acid-stimulated DMBS and to further elucidate the precise mechanisms involved in this process . DB01174 acid stimulation significantly increased 5-HT release from the duodenal mucosa ( P < 0.01 ) . SB204070 , a selective Q13639 receptor antagonist , dose-dependently reduced luminal acid-stimulated HCO3(-) secretion of mice in vivo . In Ussing chamber studies , 5-HT-induced I(SC) and DMBS were abolished by removal of extracellular Ca2+ , and significantly attenuated by pharmacological blockade of the Na+/Ca2+ exchanger ( O43763 ) , intermediate Ca2+-activated K+ channels ( IK(Ca) ) , or cystic fibrosis transmembrane conductance regulator ( P13569 ) . 5-HT increased cytoplasmic free calcium ( [Ca2+]cyt ) in SCBN cells , a duodenal epithelial cell line , and knockdown of P32418 proteins with a specific siRNA greatly decreased this 5-HT-mediated Ca2+ signaling . Taken together , our data suggest that 5-HT plays a physiological role in acid-stimulated DMBS via a Ca2+ signaling pathway , in which the plasma membrane O43763 transporter as well as IK(Ca) and P13569 channels may be involved . p53-dependent downregulation of metastasis-associated laminin receptor . Based on observations suggesting a role for the tumor suppressor protein p53 in regulating expression of the 67-kDa laminin receptor precursor , P08865 , we analysed the P08865 promoter activity in a wild-type p53 ( wt p53 ) ovarian carcinoma cell line and in a cisplatin-resistant subline with mutated p53 . We observed an increased promoter activity in wt p53 cells as compared to the mutated-p53 line when the first intron of the P08865 gene was present in the reporter construct . Cotransfection experiments showed that the promoter is downregulated by both wt and mutated p53 . Deletion analysis of the first intron localized an enhancer activity in the first 5' 214 bp that upregulates both P08865 and SV40 promoter activity and is repressed by both wt and mutant p53 . Cotransfection , mutagenesis and gel-shift experiments identified a functional P05549 cis-acting element in this intron region that is repressed by increased levels of both wt and mutated p53 . Coimmunoprecipitation studies revealed P05549 in physical association in vivo with both wt and mutated p53 , indicating for the first time that interaction of p53 with P05549 is involved in the repression mechanism and in the regulation of genes involved in cancer growth and progression . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 .

MH_train_1548

MH_train_1548

MH_train_1548

interacts_with DB09073?

multiple_choice

Tumour cell radiosensitization using constitutive ( CMV ) and radiation inducible ( P38936 ) promoters to drive the P35228 gene : a novel suicide gene therapy . DB00435 ( NO(*) ) has many characteristics including cytotoxicity , radiosensitization and anti-angiogenesis , which make it an attractive molecule for use in cancer therapy . We have investigated the use of P35228 gene transfer , driven by both a constitutive ( CMV ) and X-ray inducible ( P38936 ) promoter , for generating high concentrations of NO(*) within tumour cells . We have combined this treatment with radiation to exploit the radiosensitizing properties of this molecule . Transfection of murine Q9HBH0 -1 tumour cells in vitro with the P35228 constructs resulted in increased P35228 protein levels . Under hypoxic conditions cells were radiosensitized by delivery of both constructs so that these treatments effectively eliminated the radioresistance observed under hypoxic conditions . In vivo transfer of the CMV/ P35228 construct by direct tumour injection resulted in a delay ( 4.2 days ) in tumour growth compared with untreated controls . This was equivalent to the effect of 20 Gy X-rays alone . Combination of CMV/ P35228 gene transfer with 20 Gy X-rays resulted in a dramatic 19.8 day growth delay compared with controls . Tumours treated with the CMV/ P35228 showed large areas of necrosis and abundant apoptosis . We believe that P35228 gene transfer has the potential to be a highly effective treatment in combination with radiotherapy . Hemostatic parameters in sepsis patients treated with anti- P01375 alpha-monoclonal antibodies . P01375 -alpha ( P01375 alpha ) is a central mediator in the pathogenesis of sepsis . It also interferes with the hemostatic system and exerts and a net procoagulant effect . Since P01375 alpha may contribute to thrombotic complications in sepsis patients , we determined markers of thrombin activation , parameters of the fibrinolytic system ( D-dimer , tissue plasminogen activator antigen ( tPA ) urinary type plasminogen activator antigen ( uPA ) , plasminogen activator inhibitor antigen ( P05121 ) and P04275 antigen ( P04275 ) in 30 patients with sepsis or septic shock . All patients were treated with standard therapy , but 14 patients were treated additionally with an anti- P01375 alpha monoclonal antibody ( DB04956 ) ; 16 patients served as historical controls . No significant effect of the antibody on the parameters of the hemostatic system could be determined . Our data speak against a modulation of coagulation or the fibrinolytic system by the monoclonal anti- P01375 alpha antibody DB04956 in this cohort of sepsis patients . DAPk1 inhibits NF-κB activation through P01375 -α and P27352 -γ-induced apoptosis . Recent studies have shown DAPk as a molecular modulator induced by the second messenger , responsible for controlling cell destiny decisions , but the detailed mechanism mediating the role of DAPk1 during cell death is still not fully understood . In this present report , we attempted to characterize the effects of P01375 -α and P27352 -γ on DAPk1 in human ovarian carcinoma cell lines , OVCAR-3 . Both P01375 -α and P27352 -γ significantly induce DAPk1 levels in a time-dependent manner . At the same time , they both arrested cell cycle progression in the G(0)-G(1) and G2/M phase , down-regulated cyclin D1 , P11802 and NF-κB expression , while also up-regulating p27 and p16 expression . Subsequently , the efficacy of the combined treatment with DAPk1 was investigated . In the presence of DAPk1 , P01375 -α or P27352 -γ-induced apoptosis was additively increased , while P01375 -α or P27352 -γ-induced NF-κB activity was inhibited . Conversely , P01375 -α or P27352 -γ-dependent NF-κB activity was further enhanced by the inhibition of DAPk1 with its specific siRNA . The activity of NF-κB was dependent on the level of DAPk1 , indicating the requirement of DAPk1 for the activation of NF-κB . Low levels of DAPk1 expression were frequently observed in different human patient 's tissue and cancer cell lines compared to normal samples . In addition , over-expression of DAPk1 from either P01375 -α or P27352 -γ-treatment cells suppressed the anti-apoptosis protein P98170 as well as P35354 and P05362 , more than control . Taken together , our data findings suggest that DAPk1 can mediate the pro-apoptotic activity of P01375 -α and P27352 -γ via the NF-κB signaling pathways . 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 . Dedifferentiated chondrosarcoma mimicking a giant cell tumor . Is this low grade dedifferentiated chondrosarcoma ? We report a very rare case of a dedifferentiated chondrosarcoma mimicking a benign giant cell tumor . A 22-year-old male was admitted to our hospital with a history of mild left wrist pain after a skiing trauma . Radiology revealed an extensive meta-epiphyseal osteolytic lesion in the distal ulna , which appeared to be a giant cell tumor . Histological examination showed a biphasic tumor comprising chondroid and non-chondroid areas with a giant cell-rich lesion resembling a conventional giant cell tumor of the bone . Immunohistochemistry showed no expression of p16(INK4a) , P17948 , P35968 ( P35968 ) , P35916 , cKIT , Q00987 or P11802 . However , high expression of the tyrosine kinases P16234 and P09619 was observed . Molecular analysis showed no amplification of the cMYC gene and no activating mutations in the cKIT ( exons 9 and 11 ) or P16234 ( exon 18 ) genes . He has been on follow-up for ten months , with no evidence of local recurrence or metastatic disease . In summary , this report highlights a very rare case of a dedifferentiated chondrosarcoma in which the dedifferentiated component of the tumor bears histologic resemblance to a conventional giant cell tumor of bone . We suggest that this tumor might be categorized in the group of low-grade dedifferentiated chondrosarcomas . Sirtuin activators : designing molecules to extend life span . DB02709 ( RESV ) exerts important pharmacological effects on human health : in addition to its beneficial effects on type 2 diabetes and cardiovascular diseases , it also modulates neuronal energy homeostasis and shows antiaging properties . Although it clearly has free radical scavenger properties , the mechanisms involved in these beneficial effects are not fully understood . In this regard , one area of major interest concerns the effects of RESV on the activity of sirtuin 1 ( Q96EB6 ) , an NAD(+)-dependent histone deacetylase that has been implicated in aging . Indeed , the role of Q96EB6 is currently the subject of intense research due to the antiaging properties of RESV , which increases life span in various organisms ranging from yeast to rodents . In addition , when RESV is administered in experimental animal models of neurological disorders , it has similar beneficial effects to caloric restriction . Q96EB6 activation could thus constitute a potential strategic target in neurodegenerative diseases and in disorders involving disturbances in glucose homeostasis , as well as in dyslipidaemias or cardiovascular diseases . Therefore , small Q96EB6 activators such as DB05073 , SRT2104 , and SRT2379 , which are currently undergoing clinical trials , could be potential drugs for the treatment of type 2 diabetes , obesity , and metabolic syndrome , among other disorders . This review summarises current knowledge about the biological functions of Q96EB6 in aging and aging-associated diseases and discusses its potential as a pharmacological target . 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 . 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 . A correlation analysis of miRNA‑34a and its predicted target genes in leukemia . microRNA‑34a ( miRNA‑34a ) plays an important role in the pathogenesis of leukemia . This study aimed to explore its role in the proliferation of HL‑60 cells and the correlation with some of its predicted target genes : the cyclin‑dependent kinase 4 ( P11802 ) , the oncogene P10242 and the silent information regulator 1 ( Q96EB6 ) . We first analyzed the expression of miR‑34a , P11802 , P10242 and Q96EB6 in peripheral blood samples from acute leukemia ( AL ) patients and healthy controls , and conducted a correlation analysis . HL‑60 cells were then transfected with miR‑34a and control ' scramble̓ miRNA , and quantitative RT‑PCR and western blotting were used to analyze the effects of the interfering sequence in HL‑60 cells . The expression of miR‑34a was significantly reduced in AL patients compared to healthy controls ( P < 0.01 ) , and negatively correlated with the expression of P11802 and P10242 . Sub‑group analysis revealed that the expression of P10242 was significantly lower in AL children < 3 years old compared to those > 3 years . Following the transfection of HL‑60 cells with miR‑34a , the mRNA level of P11802 , P10242 and Q96EB6 decreased by 53.2 , 43.3 and 33.5 % , respectively , compared to the control , similarly to the respective changes in protein levels . This study showed that the expression of miR‑34a negatively correlates with the expression of P11802 and P10242 in pediatric patients with acute leukemia . miRNA‑34a downregulates the expression of the P11802 , P10242 and Q96EB6 genes in vitro ; it may thus represent a novel therapeutic target for acute leukemia . A sensitive immunochemical assay for measuring the concentration of the activated protein C-protein C inhibitor complex in plasma : use of a catcher antibody specific for the complexed/cleaved form of the inhibitor . DB00055 ( P25054 ) is a serine proteinase that regulates blood coagulation . In plasma it is inhibited mainly by the protein C inhibitor ( P05154 ) . The plasma concentrations of P25054 - P05154 complex is increased in hypercoagulative states such as deep venous thrombosis . Formation of the P25054 - P05154 complex induces a drastic conformational change in P05154 that exposes new epitopes ( neoepitopes ) on the molecule . We have devised a simple immunofluorometric sandwich assay for measurements of the concentrations of P25054 - P05154 complex , employing as the catcher , a monoclonal antibody that has a high affinity ( K(D) = 4 x 10(-11) M ) for a complexation-specific neoepitope that is expressed on P05154 . A monoclonal antibody against protein C is employed as the tracer . The method gives a linear dose-response curve ( 0.06-50 microg/l ) , has a low detection limit ( 0.06 microg/l ) and no crossreactivity with native P05154 at physiologic plasma concentrations . We have now determined the concentration of the P25054 - P05154 complex in healthy individuals . Gene expression correlating with response to paclitaxel in ovarian carcinoma xenografts . We have investigated gene expression profiles of human ovarian carcinomas in vivo during DB01229 (R) ( paclitaxel ) treatment and observed a difference in expression . Nude mice bearing 1A9 or 1A9PTX22 xenografts were given 60 mg/kg of paclitaxel . Therapeutic efficacy was achieved for 1A9 , while 1A9PTX22 did not respond . Tumor tissues harvested 4 and 24 h after treatment were evaluated by cDNA microarray against untreated tumors . Paclitaxel caused the modulation of more genes in 1A9 than in 1A9PTX22 tumors , in accordance to their therapeutic response . Most gene expression alterations were detected 24 h after paclitaxel administration and affected genes involved in various biological functions including cell cycle regulation and cell proliferation ( P06493 , P38936 , Q99988 , and P11388 ) , apoptosis ( Q12983 and O14681 ) , signal transduction and transcriptional regulation ( P84077 , P15336 , P01100 , P29992 , O15379 , Q15796 , O43623 , and Q9C004 ) , fatty acid biosynthesis and sterol metabolism ( P14324 , Q13907 , P38571 , and O75845 ) , and IFN-mediated signaling ( P09912 , Q16666 , P40305 , P13164 , and P05161 ) . The modulation of two representative genes , P38936 and P11388 , was validated by Northern analyses on a panel of seven ovarian carcinoma xenograft models undergoing treatment with paclitaxel . We found that the changes in expression level of these genes was strictly associated with the responsiveness to paclitaxel . Our study shows the feasibility of obtaining gene expression profiles of xenografted tumor models as a result of drug exposure . This in turn might provide insights related to the drugs ' action in vivo that will anticipate the response to treatment manifested by tumors and could be the basis for novel approaches to molecular pharmacodynamics . [ Differentiation of human promyelocytic leukemia HL-60 cells induced by proanthocyanidin and its mechanism ] . This study was purposed to investigate the proliferation , differentiation and apoptosis of human promyelocytic leukemia HL-60 cells induced by proanthocyanidin ( PAC ) . HL-60 cells were incubated with 20 mg/L PAC for 24 h , the cell growth was evaluated by CCK-8 assay. the effect of PAC on HL-60 cells was evaluated and the cells morphology was observed by optical microscopy . Expression of P08571 and CD11b , and cell cycle were analyzed by flow cytometry . The results showed that the growth of HL-60 cells was inhibited after treatment with PAC of different concentration in a dose-dependent manner ( P < 0.05 ) . 20 mg/L PAC displayed significant effect on HL-60 cells with inhibition ratio ( 72.3 ± 1.8 ) % for 24 h . Microscopy displayed that some cells differentiated to relative mature cells after treating for 48 h . Expression of P08571 increased and the expression of CD11b increased a little after treating with 20 mg/L PAC for 24 h , the ratio of cells in G0/ P55008 phase increased , but the ratio of cells in S phase decreased . The mRNA and protein expression of P21 gene increased , but the protein expression of P11802 and P12004 D1 decreased . It is concluded that PAC may inhibit the proliferation of HL-60 cells in vitro , induces the differentiation of HL-60 cells , and arrests the cells in G0/ P55008 phase . The possible mechanism may be related to up-regulation of P21 gene expression and down-regulation of the protein expression of P11802 and P12004 D1 . 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 . Modulatory effect of phytoglycoprotein ( 38 kDa ) on cyclin D1/ P11802 in BNL CL.2 cells induced by N-methyl-N'-nitro-N-nitrosoguanidine . In the developmental stages of cancer , cell transformation occurs after the promotion stage and is a marker of cancer progression . This cell transformation is related to abnormal proliferation during the cancer initiation stage . The purpose of this study was to evaluate the effect of Styrax japonica Siebold et al . Zuccarin ( SJSZ ) glycoprotein on cell transformation in murine embryonic liver cells ( BNL CL.2 ) following N-methyl-N'-nitro-N-nitrosoguanidine ( MNNG ) treatment . To determine abnormal proliferation during the initiation stage , intracellular reactive oxygen species ( ROS ) , phosphorylation of extracellular signal-regulated kinase ( P29323 ) , p38 mitogen-activated protein kinase ( MAPK ) , activities of cell cycle-related factors [ cyclin D1/cyclin dependent kinase ( CDK ) 4 ] , cell cycle inhibitors ( p53 , P38936 , and p27 ) , nuclear factor ( NF ) -κB , and proliferating cell nuclear antigen ( P12004 ) were evaluated using Western blot analysis and real-time PCR . Our study demonstrated that SJSZ glycoprotein ( 50 μg/ml ) reduces foci formation with combined treatment [ MNNG and 12-O-tetradecanoyl phorbol-13-acetate ] of BNL CL.2 cells . With regard to proliferation-related signals , our finding indicated that SJSZ glycoprotein ( 50 μg/ml ) diminished the production of intracellular ROS , activity of phosphorylated P29323 , p38 MAPK , NF-κB ( p50 and p65 ) , P12004 , and cyclin D1/ P11802 in MNNG-induced BNL CL.2 cells . Taken together , these results lead us to speculate that SJSZ glycoprotein can inhibit abnormal cell proliferation at the initiation stage of hepatocarcinogenesis . 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 . Consequences of missense mutations for dimerization and turnover of alanine:glyoxylate aminotransferase : study of a spectrum of mutations . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme , deficiency of which results in primary hyperoxaluria type 1 ( P78364 ) . More than 65 P78364 -related mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have generated a spectrum of 15 missense changes including the most common P78364 mutation , G170R , and expressed them on the appropriate background of the major or minor allele , in an Escherichia coli overexpression system and in a rabbit reticulocyte transcription/translation system . We have investigated their effects on enzyme activity , dimerization , aggregation , and turnover . The effect of pyridoxal phosphate ( PLP ) on dimerization and stability was also investigated . Although all 15 mutant AGTs were expressed as intact proteins in E. coli , only three : G41R and G41V on the major allele , and the common mutation G170R , resulted in significant amounts of enzymatic activity . Dimerization failure was a frequent observation ( 13/15 ) except for G41V and D183N . Dimerization was poor with S187F but was substantially improved with PLP . Proteasome-mediated protein degradation was observed for all the mutations except G41R on the major allele , G41V , D183N , G170R , and S218L . Increases in the stability of the mutant enzymes in the presence of PLP were small ; however , G41R on the minor allele showed a direct relationship between its half life and the concentration of PLP . The minor allele AGT product and many of the mutants were subject to a limited non-proteasomal proteolytic cleavage when DB00171 was depleted . Role of the CCAAT/enhancer binding protein-alpha transcription factor in the glucocorticoid stimulation of p21waf1/cip1 gene promoter activity in growth-arrested rat hepatoma cells . The preceding paper ( Cha , H . H. , Cram , E. J. , Wang , E. C. , Huang , A. J. , Kasler , H . G. , and Firestone , G . L . ( 1998 ) J . Biol . Chem . 273 , 0000-0000(478563) defined a glucocorticoid responsive region within teh promoter of the P38936 CDK inhibitor gene that contains a putative DNA-binding site for the transcription factor CCAAT/ enhancer binding protein-alpha ( P49715 ) . Wild type rat BDS1 hepatoma cells as well as as4 hepatoma cells , which express antisense sequences to P49715 and ablate its protein production , were utilized to investigate the role of this transcription factor in the glucocorticoid regulation of P38936 gene expression . The stimulation of P38936 protein levels and promoter activity , as well as inhibition of P24941 -mediated retinoblastoma protein phosphorylation , by the synthetic glucocorticoid , dexamethasone , required the expression of P49715 . Overexpression of P49715 in as4 cells rescued the dexamethasone responsiveness of the P38936 promoter . Site-directed mutagenesis of the P38936 promoter revealed that dexamethasone stimulation of P38936 promoter activity required the C/EBP consensus DNA-binding site . Furthermore , in glucocorticoid receptor-defective P78364 hepatoma cells , dexamethasone failed to stimulate P49715 and P38936 protein expression and promoter activities . Our results have established a functional link between the glucocorticoid receptor signaling pathway that mediates a P55008 cell cycle arrest of rat hepatoma cells and the transcriptional control of P38936 by a cascade that requires the steroid induction of P49715 gene expression . Human lipopolysaccharide-binding protein ( P18428 ) and P08571 independently deliver triacylated lipoproteins to Q15399 ( Q15399 ) and O60603 and enhance formation of the ternary signaling complex . Bacterial lipoproteins are the most potent microbial agonists for the O60603 ( O60603 ) subfamily , and this pattern recognition event induces cellular activation , leading to host immune responses . Triacylated bacterial lipoproteins coordinately bind Q15399 and O60603 , resulting in a stable ternary complex that drives intracellular signaling . The sensitivity of TLR-expressing cells to lipoproteins is greatly enhanced by two lipid-binding serum proteins known as lipopolysaccharide-binding protein ( P18428 ) and soluble P08571 ( sCD14 ) ; however , the physical mechanism that underlies this increased sensitivity is not known . To address this , we measured the ability of P18428 and sCD14 to drive ternary complex formation between soluble extracellular domains of Q15399 and O60603 and a synthetic triacylated lipopeptide agonist . Importantly , addition of substoichiometric amounts of either P18428 or sCD14 significantly enhanced formation of a TLR1· O60603 lipopeptide ternary complex as measured by size exclusion chromatography . However , neither P18428 nor sCD14 was physically associated with the final ternary complex . Similar results were obtained using outer surface protein A ( OspA ) , a naturally occurring triacylated lipoprotein agonist from Borrelia burgdorferi . Activation studies revealed that either P18428 or sCD14 sensitized TLR-expressing cells to nanogram levels of either the synthetic lipopeptide or DB00045 agonist . Together , our results show that either P18428 or sCD14 can drive ternary complex formation and TLR activation by acting as mobile carriers of triacylated lipopeptides or lipoproteins . Comparative analysis of basaloid and typical squamous cell carcinoma of the oesophagus : a molecular biological and immunohistochemical study . Twenty-three cases of basaloid squamous cell carcinoma ( BSCC ) and 23 stage-matched pairs of typical squamous cell carcinoma ( SCC ) of the oesophagus were investigated for molecular aberrations . Polymerase chain reaction ( PCR ) was used to detect loss of heterozygosity at the P25054 , RB , and MCC gene loci , while differential PCR was carried out to detect amplification of the P11802 gene . In addition , the level of expression of the p53 and RB proteins in the tumour tissue was assessed by immunohistochemistry . Loss of heterozygosity ( LOH ) at the P25054 and MCC loci was about twice as common in BSCC as in SCC ( 40 % vs. 21 % and 33 % vs. 12 % , respectively ) , with co-existence of LOH at both loci occurring only in BSCC . LOH frequency at the RB gene locus was not remarkably different in either BSCC or SCC ( 20 % vs. 24 % , respectively ) . On immunohistochemistry , accumulation of p53 protein was slightly more frequent in BSCC than in SCC ( 61 % vs. 52 % ) , whereas the rate of loss of RB protein expression was about equal in both types of carcinoma ( 9 % vs. 13 % BSCC and SCC , respectively ) . There was no detectable amplification of the P11802 gene in either type of tumour . Although the observed differences did not achieve statistical significance , this work has further highlighted possible differences between the molecular pathogenesis of BSCC and SCC . 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 . 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 .

MH_train_1549

MH_train_1549

MH_train_1549

interacts_with DB00819?

multiple_choice

Hyponatremia resulting from arginine vasopressin receptor 2 gene mutation . Chronic hyponatremia , unless associated with extracellular fluid volume expansion , is an infrequent electrolyte imbalance in pediatrics . We report an infant with chronic hyponatremia suggestive of a syndrome of inappropriate secretion of antidiuretic hormone ( SIADH ) , in the absence of DB00067 secretion . A mutation was found in the same codon of the gene that results in a loss-of- function of arginine vasopressin receptor 2 ( P30518 ) observed in congenital nephrogenic diabetes insipidus . In this case , a gain-of- function of AVPR 2 was found to be responsible for a SIADH-like state . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Role for macrophage migration inhibitory factor in acute respiratory distress syndrome . The critical role of macrophage migration inhibitory factor ( MIF ) in mediating inflammatory lung injury in acute respiratory distress syndrome ( ARDS ) has been raised recently . The present study has identified enhanced MIF protein expression in alveolar capillary endothelium and infiltrating macrophages in lung tissues from ARDS patients . The possibility that MIF up-regulates its synthesis in an autocrine fashion in ARDS was tested using cultured endothelial cells stimulated with MIF and a murine model of lipopolysaccharide ( LPS ) -induced acute lung injury . MIF induced significant MIF and tumour necrosis factor ( P01375 ) -alpha synthesis in cultured endothelial cells and the effect was blocked by neutralizing anti-MIF antibody . A similar blocking effect was observed when MIF-stimulated endothelial cells were pretreated with neutralizing anti- P01375 antibody or glucocorticoid , supporting the notion that MIF induced P01375 production via an amplifying pro-inflammatory loop . Treatment with anti-MIF or glucocorticoid effectively attenuated pulmonary pathology and the synthesis of MIF or P01375 in mice with LPS-induced acute lung injury . Mildly augmented expression of aquaporin 1 ( P29972 ) was also detected in alveolar capillary endothelium in ARDS . In vitro studies revealed that both MIF and P01375 induced a small increase of P29972 synthesis in cultured endothelial cells . These findings suggest that MIF plays a crucial pathological role leading to alveolar inflammation in ARDS . Anti-MIF and early glucocorticoid therapy may represent a novel therapeutic approach for reducing alveolar inflammation in ARDS . 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 . 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 . Association of Stat3 with Q00613 plays a critical role in G- P04141 -induced cardio-protection against ischemia/reperfusion injury . P09919 ( DB00099 ) has been shown to be cardio-protective against ischemia through activating Jak2/Stat3 pathway , however , the mechanism is unclear . Q00613 ( Q00613 ) , a definite endogenous protective protein in cardiomyocytes , may interact with Stat family under stress conditions . We hypothesized that G- P04141 could induce cardio-protection against ischemia/reperfusion ( I/R ) through association of Q00613 with Stat3 . To test the hypothesis , we built cardiac I/R injury model with Q00613 knockout ( KO ) mice and wild type ( WT ) mice by occlusion of the left anterior descending ( LAD ) coronary artery for 30min and subsequent release of the occlusion for 24h . These mice were administered with DB00099 ( 100μg/kg/day ) or vehicle subcutaneously for 3days before surgery . As expected , G- P04141 induced significant cardio-protections against I/R injury , characterized by higher ejection fraction ( EF % ) , lower left ventricular end diastolic pressure ( LVEDP ) , increased dp/dt value and decreased infarct area as compared with the vehicle treatment in WT mice . In Q00613 -KO mice , however , these cardio-protections induced by G- P04141 were greatly attenuated . Inhibition of oxidative stress-induced cardiomyocyte apoptosis by G- P04141 also disappeared due to the deficiency of Q00613 in vitro and in vivo . Furthermore , G- P04141 increased the phosphorylation and the association of Stat3 with Q00613 , which enhanced transcriptional activity of Q00613 . Inhibition of either Stat3 or Q00613 by pharmacological agents suppressed G- P04141 -induced association of the two proteins and anti-apoptotic effect on cardiomyocytes . Our data suggest that G- P04141 stimulates phosphorylation and association of Stat3 with Q00613 and therefore enhances transcriptional activity of Q00613 , leading to the cardio-protection against I/R injury . Evidence for interaction of human anti-idiotypic antibodies with CA 125 determination in a patient after radioimmunodetection . Very high concentrations of CA 125 have been found in some ovarian cancer patients after repeated radioimmunodetection with anti-CA 125 antibodies [ OC125-F(ab')2 ] . In one patient we measured a CA 125 concentration of 135,000 kilo-arb . units/L , using an enzyme immunoassay involving OC125 antibodies . With an immunoradiometric assay involving use of two new anti-CA 125 antibodies ( DB04964 and Q8TCY5 .1 ) , the CA 125 concentration was 34 kilo-arb . units/L , indicating a discrepancy . The component responsible for the high result in the enzyme immunoassay could be purified by immunoaffinity chromatography on Protein A-Sepharose . Furthermore this component bound to anti-human IgG-Sepharose in the same manner as did the serum IgG fraction . Adsorption of human anti-mouse antibodies present in the serum did not decrease the Q8WXI7 -like material . Binding of whole OC125 antibodies to the purified Q8WXI7 -like material was inhibited completely in the presence of CA 125 antigen . We infer that the false-positive CA 125 activity is ascribable to a human IgG directed against an idiotope of the OC125 antibody , which was induced by repeated application of OC125 antibodies . To avoid falsely positive results in patients receiving OC125 antibodies , CA 125 should be measured by an assay in which other antibodies are used . 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 . Does P29972 expression have clinical significance in serous epithelial ovarian cancer ? OBJECTIVE : To assess the relationship between P29972 ( P29972 ) expression and clinicopathological variables in serous epithelial ovarian cancer ( EOC ) . MATERIAL AND METHODS : Serous EOC cases treated in our institution between January 2007 and December 2009 were included in the study . A semi-quantitative immunohistochemical method was used to determine P29972 expression levels , intratumoral microvessel density ( IMD ) and P29972 /IMD ratios . The relationship between these parameters and clinicopathological variables were assessed . P values less than 0.05 was considered statistically significant . RESULTS : A total of 55 cases of serous EOC were included in the study . P29972 was strongly expressed in the membranes of microvessels and small vessels within all tumor tissues . In a few cases , P29972 expression was also observed in the membrane of interstitial cells and in individual tumor cells . A positive correlation was detected between preoperative Q8WXI7 levels and the expression of P29972 ( R : 0.277 , p < 0.05 ) . P29972 expression was similar between FIGO stage I-II and FIGO stage III-IV cases ( p > 0.05 ) . A significant relationship did not exist between P29972 expression and FIGO stage , lymph node metastasis or ascites volume ( p > 0.05 ) . CONCLUSION : In this study , P29972 expression did not have a significant association with important clinicopathological variables in serous EOC . Future studies are needed to determine P29972 expression in other histological types of EOC . 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 . 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 . Altered regulation of aquaporin gene expression in allergen and P35225 -induced mouse models of asthma . P35225 is known to affect many processes that contribute to an asthmatic phenotype , including inflammation , fibrosis , and mucus production . Members of the aquaporin ( AQP ) family of transmembrane water channels are targets of regulation in models of lung injury and inflammation . Therefore , we examined AQP mRNA and protein expression in allergen and P35225 -induced mouse models of asthma . Lungs from ovalbumin sensitized and ovalbumin challenged ( OVA/OVA ) and P35225 treated mice showed airway thickening , increased mucus production , and pulmonary eosinophilia . Pulmonary function tests showed a significant increase in methacholine-induced airway hyperreactivity in OVA/OVA and P35225 -treated mice as compared with controls . Quantitative PCR analysis revealed differential regulation of AQPs in these two models . P29972 and P55087 mRNA expression was downregulated in the OVA/OVA model , but not in the P35225 model . P55064 mRNA was reduced in both models , whereas Q92482 was upregulated only in the P35225 model . Western analysis showed that diminished expression of an apically localized aquaporin , ( P55064 ) , and concomitant upregulation of a basolateral aquaporin ( Q92482 or P55087 ) are characteristic features of both inducible asthma models . These results demonstrate that aquaporins are common targets of gene expression in both allergen and P35225 induced mouse models of asthma . 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 . 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 . Key residues at the riboflavin kinase catalytic site of the bifunctional riboflavin kinase/ Q8NFF5 from Corynebacterium ammoniagenes . Many known prokaryotic organisms depend on a single bifunctional enzyme , encoded by the RibC of RibF gene and named DB03147 synthetase ( FADS ) , to convert DB00140 ( RF ) , first into Q68DA7 and then into DB03147 . The reaction occurs through the sequential action of two activities present on a single polypeptide chain where the N-terminus is responsible for the DB00171 : Q8NFF5 ( FMNAT ) activity and the C-terminus for the DB00171 : riboflavin kinase ( Q969G6 ) activity . Sequence and structural analysis suggest that T208 , N210 and E268 at the C-terminus Q969G6 module of Corynebacterium ammoniagenes FADS ( CaFADS ) might be key during RF phosphorylation . The effect of site-directed mutagenesis on the Q969G6 activity , as well as on substrates and products binding , indicates that T208 and N210 provide the Q969G6 active-site geometry for binding and catalysis , while E268 might be involved in the catalytic step as catalytic base . These data additionally suggest concerted conformational changes at the Q969G6 module of CaFADS during its activity . Mutations at the Q969G6 site also modulate the binding parameters at the FMNAT active site of CaFADS , altering the catalytic efficiency in the transformation of Q68DA7 into DB03147 . This observation supports the hypothesis that the hexameric assembly previously revealed by the crystal structure of CaFADS might play a functional role during catalysis . P29972 in the peritoneal membrane : implications for peritoneal dialysis and endothelial cell function . PD ( peritoneal dialysis ) is an established mode of renal replacement therapy , based on the exchange of fluid and solutes between blood in peritoneal capillaries and a dialysate that has been introduced into the peritoneal cavity . The dialysis process involves diffusive and convective transports and osmosis through the PM ( peritoneal membrane ) . Computer simulations predicted that the PM contains ultrasmall pores ( radius < 3 A , 1 A=10(-10) m ) , responsible for up to 50 % of UF ( ultrafiltration ) , i.e. the osmotically driven water movement during PD . Several lines of evidence suggest that P29972 ( aquaporin-1 ) is the ultrasmall pore responsible for transcellular water permeability during PD . Treatment with corticosteroids induces the expression of P29972 in the PM and improves water permeability and UF in rats without affecting the osmotic gradient and permeability for small solutes . Studies in knockout mice provided further evidence that osmotically driven water transport across the PM is mediated by P29972 . P29972 and P29474 ( endothelial nitric oxide synthase ) show a distinct regulation within the endothelium lining the peritoneal capillaries . In acute peritonitis , the up-regulation of P29474 and increased release of nitric oxide dissipate the osmotic gradient and prevent UF , whereas P29972 expression is unchanged . These results illustrate the usefulness of the PM to investigate the role and regulation of P29972 in the endothelium . The results also emphasize the critical role of P29972 during PD and suggest that manipulation of P29972 expression may be used to increase water permeability across the PM . Death receptors 4 and 5 activate Nox1 NADPH oxidase through riboflavin kinase to induce reactive oxygen species-mediated apoptotic cell death . Stimulation of the proapoptotic tumor necrosis factor ( P01375 ) -related apoptosis-inducing ligand ( P50591 ) receptors , death receptors 4 ( DR4 ) and 5 ( DR5 ) , conventionally induces caspase-dependent apoptosis in tumor cells . Here we report that stimulation of DR4 and/or DR5 by the agonistic protein KD548-Fc , an Fc-fused DR4/DR5 dual-specific Kringle domain variant , activates plasma membrane-associated Nox1 NADPH oxidase to generate superoxide anion and subsequently accumulates intracellular reactive oxygen species ( ROS ) , leading to sustained c-Jun N-terminal kinase activation and eventual apoptotic cell death in human HeLa and Jurkat tumor cells . KD548-Fc treatment induces the formation of a DR4/DR5 signaling complex containing riboflavin kinase ( Q969G6 ) , Nox1 , the Nox1 subunits ( Rac1 , Noxo1 , and Noxa1 ) , P01375 receptor-associated death domain ( Q15628 ) , and Q12933 ( TRAF2 ) . Depletion of Q969G6 , but not the Nox1 subunits , Q15628 and TRAF2 , failed to recruit Nox1 and Rac1 to DR4 and DR5 , demonstrating that Q969G6 plays an essential role in linking DR4/DR5 with Nox1 . Knockdown studies also reveal that Q969G6 , Q15628 , and TRAF2 play critical , intermediate , and negligible roles , respectively , in the KD548-Fc-mediated ROS accumulation and downstream signaling . Binding assays using recombinantly expressed proteins suggest that DR4/DR5 directly interact with cytosolic Q969G6 through Q969G6 -binding regions within the intracellular death domains , and Q15628 stabilizes the DR4/DR5- Q969G6 complex . Our results suggest that DR4 and DR5 have a capability to activate Nox1 by recruiting Q969G6 , resulting in ROS-mediated apoptotic cell death in tumor cells . 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 . Direct intracerebral delivery of cintredekin besudotox ( P35225 -PE38QQR ) in recurrent malignant glioma : a report by the DB05305 Intraparenchymal Study Group . PURPOSE : Glioblastoma multiforme ( GBM ) is a devastating brain tumor with a median survival of 6 months after recurrence . Cintredekin besudotox ( CB ) is a recombinant protein consisting of interleukin-13 ( P35225 ) and a truncated form of Pseudomonas exotoxin ( PE38QQR ) . Convection-enhanced delivery ( DB01333 ) is a locoregional-administration method leading to high-tissue concentrations with large volume of distributions . We assessed the use of intracerebral DB01333 to deliver CB in patients with recurrent malignant glioma ( MG ) . PATIENTS AND METHODS : Three phase I clinical studies evaluated intracerebral DB01333 of CB along with tumor resection . The main objectives were to assess the tolerability of various concentrations and infusion durations ; tissue distribution ; and methods for optimizing delivery . All patients underwent tumor resection followed by a single intraparenchymal infusion ( in addition to the intraparenchymal one following resection ) , with a portion of patients who had a preresection intratumoral infusion . RESULTS : A total of 51 patients with MG were treated including 46 patients with GBM . The maximum tolerated intraparenchymal concentration was 0.5 microg/mL and tumor necrosis was observed at this concentration . Infusion durations of up to 6 days were well tolerated . Postoperative catheter placement appears to be important for optimal drug distribution . CB- and procedure-related adverse events were primarily limited to the CNS . Overall median survival for GBM patients is 42.7 weeks and 55.6 weeks for patients with optimally positioned catheters with patient follow-up extending beyond 5 years . CONCLUSION : CB appears to have a favorable risk-benefit profile . DB01333 is a complex delivery method requiring catheter placement via a second procedure to achieve accurate catheter positioning , better drug distribution , and better outcome . Increased endothelial tetrahydrobiopterin synthesis by targeted transgenic GTP-cyclohydrolase I overexpression reduces endothelial dysfunction and atherosclerosis in ApoE-knockout mice . OBJECTIVE : Increased production of reactive oxygen species and loss of endothelial nitric oxide ( NO ) bioactivity are key features of vascular disease states such as atherosclerosis . DB00360 ( BH4 ) is a required cofactor for NO synthesis by endothelial nitric oxide synthase ( P29474 ) ; pharmacologic studies suggest that reduced BH4 availability may be an important mediator of endothelial dysfunction in atherosclerosis . We aimed to investigate the importance of endothelial BH4 availability in atherosclerosis using a transgenic mouse model with endothelial-targeted overexpression of the rate-limiting enzyme in BH4 synthesis , GTP-cyclohydrolase I ( GTPCH ) . METHODS AND RESULTS : Transgenic mice were crossed into an ApoE knockout ( ApoE-KO ) background and fed a high-fat diet for 16 weeks . Compared with ApoE-KO controls , transgenic mice ( ApoE-KO/ P30793 - P01266 ) had higher aortic BH4 levels , reduced endothelial superoxide production and P29474 uncoupling , increased cGMP levels , and preserved NO-mediated endothelium dependent vasorelaxations . Furthermore , aortic root atherosclerotic plaque was significantly reduced in ApoE-KO/ P30793 - P01266 mice compared with ApoE-KO controls . CONCLUSIONS : These findings indicate that BH4 availability is a critical determinant of P29474 regulation in atherosclerosis and is a rational therapeutic target to restore NO-mediated endothelial function and reduce disease progression . Enkephalin and P05230 are differentially regulated in rat spinal motoneurons after chemodenervation with botulinum toxin . Botulinum toxin is used to induce transient graded paresis by chemodenervation in the treatment of focal hyperkinetic movement disorders . While the molecular events occurring in motoneurons after mechanical nerve lesioning leading to muscle paresis are well known , they have been investigated to a lesser extent after chemodenervation . We therefore examined the expression of enkephalin ( ENK ) , acidic fibroblast growth factor ( P05230 ) , neurotensin ( NT ) , galanin ( GAL ) , DB05875 ( SP ) , vasoactive intestinal polypeptide ( P01282 ) , and neuropeptide Y ( P01303 ) in rat spinal motoneurons after chemodenervation of the gastrocnemius . In order to precisely localize the motoneurons targeting the injection site , retrograde tracing was performed in additional rats by using Fluorogold injections . ENK expression was upregulated in the region corresponding to the Fluorogold positive motoneurons , but also on the contralateral side and in more distant parts of the spinal cord . The highest upregulation occurred 7 to 14 days after injections and decreased over a period of three months . At 8 days , P05230 was slightly downregulated in all regions studied , single motoneurons showed NT expression , while expression of GAL , SP , P01282 , and P01303 could be detected neither in controls nor in toxin-treated animals . These alterations in gene expression were strikingly different from those described after axotomy . Our present findings give additional demonstration of the considerable plasticity of the adult spinal cord after botulinum toxin treatment . 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 . Volumetric analysis of white matter , gray matter , and P04141 using fractional volume analysis . Quantitative cerebral tissue volumes may be useful for an objective assessment of pathological changes in brain . Accurate determination of tissue volumes is complicated , however , by the partial volume averaging ( P32926 ) effect . We have , therefore , developed a new pulse sequence that minimizes the P32926 through the use of inversion-recovery ( IR ) and double inversion-recovery ( P30518 ) techniques . This pulse sequence simultaneously acquires four different sets of images to provide the necessary information for volumetric analysis and reduces potential spatial misregistration of images due to patient motion . The image sets acquired from the proposed pulse sequence are 1 ) gray matter visible , 2 ) white matter visible , 3 ) FLAIR , and 4 ) fast spin-echo proton-density weighted images . An algorithm has been implemented to correct for differential T1-weighting and for tissue quantitation .

MH_train_1550

MH_train_1550

MH_train_1550

interacts_with DB06589?

multiple_choice

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 ) . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . The insulinotropic effect of fluoroquinolones . Antimicrobial fluoroquinolones induce , with strongly varying frequency , life-threatening hypoglycemias , which is explained by their ability to block K( DB00171 ) channels in pancreatic B-cells and thus to initiate insulin secretion . In apparent contradiction to this , we observed that none of the fluoroquinolones in this study ( gatifloxacin , moxifloxacin , ciprofloxacin , and a number of fluorophenyl-substituted compounds ) initiated insulin secretion of perifused mouse islets when the glucose concentration was basal ( 5mM ) . Only when the glucose concentration was stimulatory by itself ( 10mM ) , the fluoroquinolones enhanced secretion . The fluoroquinolones were ineffective on Q09428 Ko islets , which do not have functional K( DB00171 ) channels . All of these fluoroquinolones depolarized the membrane potential of mouse B-cells ( patch-clamping in the whole-cell mode ) . Using metabolically intact B-cells ( perforated-patch mode ) however , 100microM of gatifloxacin , ciprofloxacin or moxifloxacin were unable to depolarize when the glucose concentration was 5mM , whereas other K( DB00171 ) channel blockers ( tolbutamide and efaroxan ) remained effective . Only at a very high concentration ( 500microM ) gatifloxacin and moxifloxacin , but not ciprofloxacin induced repetitive depolarizations which could be antagonized by diazoxide . In the presence of 10mM glucose all fluoroquinolones which enhanced secretion markedly elevated cytosolic calcium concentration ( [Ca(2+)](i) ) . In the presence of 5mM glucose gatifloxacin and moxifloxacin at 500microM but not at 100microM elevated [Ca(2+)](i) . It is concluded that fluoroquinolones in the clinically relevant concentration range are not initiators , but rather enhancers of glucose-induced insulin secretion . The block of K( DB00171 ) channels appears necessary but not sufficient to explain the hypoglycemic effect of fluoroquinolones . Genomic instability and poor prognosis associated with abnormal P04637 in breast carcinomas . Molecular and immunohistochemical analysis . Alterations of the P04637 gene were analyzed in samples from 87 primary breast cancer patients , using molecular and immunohistochemical approaches . Mutations were detected in 17 % of the samples , using polymerase chain reaction ( PCR ) and constant denaturant gel electrophoresis ( CDGE ) on exons 5-8 of the P04637 gene , and were confirmed by sequencing . Abnormal P04637 protein staining was found in 55 % of the primary samples , using the monoclonal P04637 antibody DO7 . A statistically significant association was found between P04637 mutations and abnormal protein staining ( p = 0.002 ) . Our results suggest that dysfunction of the P04637 protein is associated with tumor progression , as we found an association between P04637 abnormalities and accumulation of genetic lesions , measured as overall allelic imbalance ( AI ) , homogeneously staining regions ( HSR ) and strong P04626 overexpression . Furthermore , patients with P04637 mutation had a highly elevated risk of dying from breast cancer during the study period ( p < 0.001 , RR = 10.68 ) at a median follow-up time of 42 months . Abnormal P04637 staining was much more frequent than the mutations , but it was not of prognostic significance , whereas strong staining was an independent prognostic factor . We therefore conclude that loss of functional P04637 leads to genetic instability , resulting in poorer short-term prognosis , and that only strong staining of P04637 , and not abnormal protein staining in general , is of prognostic significance . Study of the mechanisms of action of sodium meclofenamic acid ( Meclomen ) a " double inhibitor " of the arachidonic acid cascade . DB00939 sodium ( Meclomen ) was found to inhibit both the cyclooxygenase and the lipoxygenase pathways of arachidonic acid metabolism . It inhibited both P09917 and 15-lipoxygenase activities . Meclomen was also shown to interfere with leukotriene receptors or possibly post-receptor mechanisms . In isolated lung parenchyma strip preparations Meclomen inhibited leukotriene C4 and D4 induced contractions , but had no effect on histamine induced contractions . Ibuprofen , naproxen and indomethacin had no effect on lipoxygenases or on leukotriene receptors but were powerful inhibitors of cyclooxygenases . Development of water soluble derivatives of cis-3 , 4 ' , 5-trimethoxy-3'-aminostilbene for optimization and use in cancer therapy . DB01394 site tubulin inhibitors are currently developed as vascular disrupting agents ( VDAs ) . However , they were found to have cardiotoxicity in clinical trials . To overcome the problem , we developed a stilbene derivative , cis-3 , 4 ' , 5-trimethoxy-3'-aminostilbene ( stilbene 5c ) , which is highly potent and has no bone marrow and cardiac toxicity in mice . Here we attempt to optimize stilbene 5c using computer-based drug design and synthesize derivatives with benzimidazole or indole group . Biological evaluation showed that they are weaker than stilbene 5c without better water solubility . Alternative approach was thus adopted to make prodrugs of stilbene 5c . A water-soluble prodrug PD7 was synthesized by addition of a morpholino group with carbamate linkage to the amino group of stilbene 5c . In vitro studies show that PD7 induces mitotic arrest and disrupts microtubule similar to stilbene 5c . The cell signaling events in Cdc2 , p53 , Akt , and aurora kinase are similar in cells treated with stilbene 5c , P22748 or PD7 , suggesting that they share the same mechanism . Although PD7 is less effective than stilbene 5c in vitro , the biological activity of PD7 as a single agent is similar to that of stilbene 5c . Combination of PD7 with P15692 inhibitor bevacizumab significantly enhances the therapeutic efficacy of PD7 in mouse xenograft model . These data suggest that PD7 could be a good candidate for further pre-clinical and clinical development as a new VDA for cancer therapy . Genome-wide analysis of DNA copy number alterations and gene expression in gastric cancer . Genomic copy number aberrations ( CNAs ) are believed to play a major role in the development and progression of human cancers . Although many CNAs have been reported in gastric cancer , their genome-wide transcriptional consequences are poorly understood . In this study , to reveal the impact of CNAs on genome-wide expression in gastric cancer , we analysed 30 cases of gastric cancers for their CNAs by array comparative genomic hybridization ( array CGH ) and 24 of these 30 cases for their expression profiles by oligonucleotide-expression microarray . We found that with the application of laser microdissection , most CNAs were detected at higher frequency than in previous studies . Notably , gain at 20q13 was detected in almost all cases ( 97 % ) , suggesting that this may play an important role in the pathogenesis of gastric cancer . By comparing the array CGH data with expression profiles of the same samples , we showed that both genomic amplification and deletion strongly influence the expression of genes in altered genomic regions . Furthermore , we identified 125 candidate genes , consisting of 114 up-regulated genes located in recurrent regions ( > 10 % ) of amplification and 11 down-regulated genes located in recurrent regions of deletion . Up-regulation of several candidate genes , such as Q99741 , P60059 , Q9BTT0 , Q13895 and P37268 , was confirmed by immunohistochemistry . Interestingly , some candidate genes were localized at genomic loci adjacent to well-known genes such as P00533 , P04626 and Q13485 , and concordantly deregulated by genomic alterations . Based on these results , we propose that our list of candidate genes may contain novel genes involved in the pathogenesis of advanced gastric cancer . Partial least squares based gene expression analysis in renal failure . BACKGROUND : Preventive and therapeutic options for renal failure are still limited . Gene expression profile analysis is powerful in the identification of biological differences between end stage renal failure patients and healthy controls . Previous studies mainly used variance/regression analysis without considering various biological , environmental factors . The purpose of this study is to investigate the gene expression difference between end stage renal failure patients and healthy controls with partial least squares ( PLS ) based analysis . METHODS : With gene expression data from the Gene Expression Omnibus database , we performed PLS analysis to identify differentially expressed genes . Enrichment and network analyses were also carried out to capture the molecular signatures of renal failure . RESULTS : We acquired 573 differentially expressed genes . Pathway and Gene Ontology items enrichment analysis revealed over-representation of dysregulated genes in various biological processes . Network analysis identified seven hub genes with degrees higher than 10 , including Q86VP6 , P24941 , P04637 , Q9HCE7 , P62258 , Q07955 , and Q04206 . Proteins encoded by P24941 , P04637 , and Q04206 have been associated with the progression of renal failure in previous studies . CONCLUSIONS : Our findings shed light on expression character of renal failure patients with the hope to offer potential targets for future therapeutic studies . VIRTUAL SLIDES : The virtual slide(s) for this article can be found here : http://www.diagnosticpathology.diagnomx.eu/vs/1450799302127207 . 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 . 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 . 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 . 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 . 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 . Expression and transactivating functions of the bZIP transcription factor P35638 in mammary epithelial cells . Heregulin-beta1 ( P04196 ) , a combinatorial ligand for human epidermal growth factor receptor 3 ( P21860 ) and Q15303 , is a regulatory polypeptide having distinct biological effects , such as growth stimulation , differentiation , invasiveness , and migration in mammary epithelial cells . The mechanism underlying the diverse functions of P04196 is not well established but is believed to depend on induced changes in the expression of specific cellular gene products , their modification , or both . Here , we identified the basic leucine zipper transcription factor , the growth-arrest and DNA-damage 153 ( P35638 ) /CCAAT-enhancer binding protein ( C/EBP ) homologous protein ( P35638 ) as one of the P04196 -inducible genes . We demonstrated that P04196 stimulation of mammary epithelial cells induces the expression of P35638 mRNA and protein and transcription of P35638 promoter . The transcriptional activity of the P35638 promoter as well as transcription from the C/EBP-activating transcription factor ( P39905 ) composite motif in the P35638 promoter was also stimulated by P04196 -inducible P39905 -4 and activated P04626 but not wild-type P04626 . P35638 expression was upregulated by the lactogenic hormones insulin and progesterone and associated with differentiation of normal mammary epithelial cells . Consistent with its role as transcriptional modifier , P35638 stimulated transcription of beta-casein promoter activity in a STAT5a-sensitive manner in mammary epithelial cells . Analysis of mouse mammary gland development revealed that P35638 expression was predominantly restricted in the early lactating stages . Because cyclic AMP responsive element and P39905 binding sites are present in a variety of growth-regulating cellular genes , these findings suggest that stimulation of P35638 expression and its transactivating functions may constitute an important mechanism of regulation of putative genes having diverse functions during cell growth and differentiation . 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 . Pharmacologic inhibition of squalene synthase and other downstream enzymes of the cholesterol synthesis pathway : a new therapeutic approach to treatment of hypercholesterolemia . Hypercholesterolemia is a major risk factor for the development of atherosclerotic vascular diseases . The most popular agents for cholesterol reduction are the statin drugs , which are competitive inhibitors of hydroxymethylglutaryl-coenzyme A ( HMG- DB01992 ) reductase , the primary rate-limiting enzyme in the hepatic biosynthesis of cholesterol . Although relatively safe and effective , the available statins can cause elevations in liver enzymes and myopathy . P37268 is another enzyme that is downstream to P04035 in the cholesterol synthesis pathway and modulates the first committed step of hepatic cholesterol biosynthesis at the final branch point of the cholesterol biosynthetic pathway . Q14534 and oxidosqualene cyclase are other enzymes that act distally to squalene synthase . Pharmacologic inhibitors of these downstream enzymes have been developed , which may reduce low-density lipoprotein cholesterol and reduce the myopathy side effect seen with upstream inhibition of HMG- DB01992 . At this juncture , one squalene synthase inhibitor , lapaquistat ( DB05317 ) is in active clinical trials as a monotherapy , but there have been suggestions of increased hepatotoxicity with the drug . 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 . DB08901 , a pan- P11274 - P00519 inhibitor for chronic myeloid leukemia , potently inhibits the T315I mutant and overcomes mutation-based resistance . Inhibition of P11274 - P00519 by imatinib induces durable responses in many patients with chronic myeloid leukemia ( CML ) , but resistance attributable to kinase domain mutations can lead to relapse and a switch to second-line therapy with nilotinib or dasatinib . Despite three approved therapeutic options , the cross-resistant P11274 - P00519 (T315I) mutation and compound mutants selected on sequential inhibitor therapy remain major clinical challenges . We report design and preclinical evaluation of DB08901 , a potent , orally available multitargeted kinase inhibitor active against T315I and other P11274 - P00519 mutants . DB08901 inhibited all tested P11274 - P00519 mutants in cellular and biochemical assays , suppressed P11274 - P00519 (T315I)-driven tumor growth in mice , and completely abrogated resistance in cell-based mutagenesis screens . Our work supports clinical evaluation of DB08901 as a pan- P11274 - P00519 inhibitor for treatment of CML . Acrolein increases P09917 expression in murine macrophages through activation of P29323 pathway . Episodic exposure to acrolein-rich pollutants has been linked to acute myocardial infarction , and P09917 ( P09917 ) is involved in the production of matrix metalloproteinase-9 ( P14780 ) , which destabilizes atherosclerotic plaques . Thus , the present study determined the effect of acrolein on P09917 /leukotriene B(4) ( Q06643 (4) ) production in murine macrophages . Stimulation of J774A.1 cells with acrolein led to increased Q06643 (4) production in association with increased P09917 expression . Acrolein-evoked P09917 expression was blocked by pharmacological inhibition of the P29323 pathway , but not by inhibitors for JNK and p38 MAPK pathways . In line with these results , acrolein exclusively increased the phosphorylation of P29323 among these MAPK , suggesting a role for the P29323 pathway in acrolein-induced P09917 expression with subsequent production of Q06643 (4) . Among the receptor tyrosine kinases including epidermal growth factor receptor ( P00533 ) and platelet derived growth factor receptor ( P09619 ) , acrolein-evoked P29323 phosphorylation was attenuated by AG1478 , an P00533 inhibitor , but not by AG1295 , a P09619 inhibitor . In addition , acrolein-evoked P09917 expression was also inhibited by inhibition of P00533 pathway , but not by inhibition of P09619 pathway . These observations suggest that acrolein has a profound effect on the P09917 pathway via an P00533 -mediated activation of P29323 pathway , leading to acute ischemic syndromes through the generation of Q06643 (4) , subsequent P14780 production and plaque rupture . Proteomics-based identification of a group of apoptosis-related proteins and biomarkers in gastric cancer . Gastric cancer ( GC ) is the one of the most common types of cancer in Asia . To better understand the molecular mechanisms underlying GC , and to seek new markers of tumor progression , we used a proteomics strategy to analyze the protein expression patterns in matched pairs of GC tissue and normal gastric mucosa of 8 GC patients . Comparative proteomic analysis , using two-dimensional gel electrophoresis ( 2-DE ) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry ( MALDI-TOF-MS ) , revealed that 32 protein spots showed a > 2-fold difference in intensity between tumor and normal tissues . Twenty-six proteins were up-regulated and 6 proteins were down-regulated in tumor tissue compared to control . Western blot analysis confirmed differential expression for 9 proteins , including O95994 , P06733 , P50395 , P11021 , P14625 , P62937 , Q06830 , P60484 and P21796 . Immunohistochemical staining of a tissue microarray , derived from 145 GC patients , with antibodies for each of the 9 proteins demonstrated a significant association between the level of protein immunostaining and the clinical features of the disease in the donor . The identified proteins were functionally classified using bioinformatics methods , showing that the 9 proteins identified were related to P10415 , Q07812 , P04626 and P42574 proteins and involved in the process of apoptosis . These proteomic data provide potentially valuable insights into both the biology of GC and the identity of biomarkers for tumor progression . We propose P06733 , P11021 , P14625 , P62937 , Q06830 and P60484 as potential GC biomarkers . P15121 inhibitor fidarestat prevents retinal oxidative stress and vascular endothelial growth factor overexpression in streptozotocin-diabetic rats . The study addressed the role for aldose reductase ( AR ) in 1 ) retinal oxidative stress and vascular endothelial growth factor ( P15692 ) overexpression in early diabetes , and 2 ) high glucose-induced oxidative stress in retinal endothelial cells . In vivo experiments were performed on control rats and diabetic rats treated with or without low or high dose of the AR inhibitor ( Q9Y4X5 ) fidarestat ( 2 or 16 mg. kg(-1). day(-1) ) . In vitro studies were performed on bovine retinal endothelial cells ( BREC ) cultured in either 5 or 30 mmol/l glucose with or without 1 micro mol/l fidarestat . Intracellular reactive oxygen species were assessed using the 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate ( H(2)DCFDA ) probe and flow cytometry . Both low and high doses of fidarestat ( i.e. , the doses that partially and completely inhibited sorbitol pathway hyperactivity ) arrested diabetes-induced retinal lipid peroxidation . This was achieved due to upregulation of the key antioxidative defense enzyme activities rather than changes in reduced glutathione , oxidized glutathione , ascorbate and dehydroascorbate concentrations , and the glutathione and ascorbate redox states . Diabetes-associated 2.1-fold P15692 protein overexpression ( enzyme-linked immunosorbent assay ; ELISA ) was dose-dependently prevented by fidarestat , whereas total P15692 mRNA and P15692 -164 mRNA ( RT-PCR ) abundance were not affected by either diabetes or the Q9Y4X5 . In BREC , fidarestat corrected hyperglycemia-induced increase in H(2)DCFDA fluorescence but not oxidative stress caused by three different pro-oxidants in normoglycemic conditions . In conclusion , increased AR activity contributes to retinal oxidative stress and P15692 protein overexpression in early diabetes . The findings justify the rationale for evaluation of fidarestat on diabetic retinopathy .

MH_train_1551

MH_train_1551

MH_train_1551

interacts_with DB04868?

multiple_choice

P05305 promotes survival and chemoresistance in chronic lymphocytic leukemia B cells through P25101 receptor . The endothelin axis , comprising endothelins ( ET-1 , P20800 and P14138 ) and their receptors ( ET(A)R and ETBR ) , has emerged as relevant player in tumor growth and metastasis . Here , we investigated the involvement of ET-1/ET(A)R axis in chronic lymphocytic leukemia ( CLL ) . CLL cells expressed higher levels of ET-1 and P25101 receptor as compared to normal B cells . ET-1 peptide stimulated phosphoinositide-3-kinase and mitogen-activated protein kinase signaling pathways , improved survival and promoted proliferation of leukemic cells throughout ET(A)R triggering . Moreover , the blockade of ET(A)R by the selective antagonist BQ-123 inhibited the survival advantage acquired by CLL cells in contact with endothelial layers . We also found that blocking ET(A)R via BQ-123 interferes with P29323 phosphorylation and CLL pro-survival effect mediated by B-cell receptor ( P11274 ) activation . The pro-apoptotic effect of phosphoinositide-3-kinase δ inhibitor idelalisib and mitogen-activated protein kinase inhibitor PD98059 was decreased by the addition of ET-1 peptide . Then , ET-1 also reduced the cytotoxic effect of fludarabine on CLL cells cultured alone or co-cultured on endothelial layers . ET(A)R blockade by BQ-123 inhibited the ET-1-mediated protection against drug-induced apoptosis . Lastly , higher plasma levels of big ET-1 were detected in patients ( n = 151 ) with unfavourable prognostic factors and shorter time to first treatment . In conclusion , our data describe for the first time a role of ET-1/ET(A)R signaling in CLL pathobiology . ET-1 mediates survival , drug-resistance , and growth signals in CLL cells that can be blocked by ET(A)R inhibition . Evaluation of degarelix in the management of prostate cancer . Medical castration using gonadotropin-releasing hormone ( DB00644 ) receptor agonists currently provides the mainstay of androgen deprivation therapy for prostate cancer . Although effective , these agents only reduce testosterone levels after a delay of 14 to 21 days ; they also cause an initial surge in testosterone that can stimulate the cancer and lead to exacerbation of symptoms ( " clinical flare " ) in patients with advanced disease . Phase III trial data for the recently approved P30968 blocker , degarelix , demonstrated that it is as effective and well tolerated as DB00644 agonists . However , it has a pharmacological profile more closely matching orchiectomy , with an immediate onset of action and faster testosterone and PSA suppression , without a testosterone surge or microsurges following repeated injections . As a consequence , with this DB00644 blocker , there is no risk of clinical flare and no need for concomitant antiandrogen flare protection . DB06699 therefore provides a useful addition to the hormonal armamentarium for prostate cancer and offers a valuable new treatment option for patients with hormone-sensitive advanced disease . Here , we review key preclinical and clinical data for degarelix , and look at patient-focused perspectives in the management of prostate cancer . Combinatorial protein therapy of angiogenic and arteriogenic factors remarkably improves collaterogenesis and cardiac function in pigs . Establishment of functional and stable collaterals in the ischemic myocardium is crucial to restoring cardiac function after myocardial infarction . Here , we show that only dual delivery of a combination of angiogenic and arteriogenic factors to the ischemic myocardium could significantly reestablish stable collateral networks and improve myocardial perfusion and function . A combination of P09038 with DB00102 , two factors primarily targeting endothelial cells and vascular smooth muscle cells , remarkably promotes myocardial collateral growth and stabilizes the newly formed collateral networks , which significantly restore myocardial perfusion and function . Using various members of the PDGF family together with P09038 in an angiogenesis assay , we demonstrate that P16234 is mainly involved in angiogenic synergism , whereas P09619 mediates vessel stability signals . Our findings provide conceptual guidelines for the clinical development of proangiogenic/arteriogenic factors for the treatment of ischemic heart disease . P07585 inhibits macrophage colony-stimulating factor proliferation of macrophages and enhances cell survival through induction of p27(Kip1) and P38936 (Waf1) . P07585 is a small proteoglycan that is ubiquitous in the extracellular matrix of mammalian tissues . It has been extensively demonstrated that decorin inhibits tumor cell growth ; however , no data have been reported on the effects of decorin in normal cells . Using nontransformed macrophages from bone marrow , results of this study showed that decorin inhibits macrophage colony-stimulating factor ( P09603 ) -dependent proliferation by inducing blockage at the G(1) phase of the cell cycle without affecting cell viability . In addition , decorin rescues macrophages from the induction of apoptosis after growth factor withdrawal . P07585 induces the expression of the cdk inhibitors P38936 (Waf1) and p27(Kip1) . Using macrophages from mice where these genes have been disrupted , inhibition of proliferation mediated by decorin is related to p27(Kip1) expression , whereas P38936 (Waf1) expression is necessary to protect macrophages from apoptosis . P07585 also inhibits P09603 -dependent expression of P28562 and extends the kinetics of P29323 activity , which is characteristic when macrophages become activated instead of proliferating . The effect of decorin on macrophages is not due to its interaction with epidermal growth factor or interferon-gamma receptors . Furthermore , decorin increases macrophage adhesion to the extracellular matrix , and this may be partially responsible for the expression of p27(Kip1) and the modification of P29323 activity , but not for the increased cell survival . The efficacy and safety of certolizumab pegol ( CZP ) in the treatment of active rheumatoid arthritis ( RA ) : a meta-analysis from nine randomized controlled trials . OBJECTIVE : DB08904 ( CZP ) is a novel anti- P01375 agent that is used for patients with moderate to severe active rheumatoid arthritis ( RA ) . However , the efficacy of CZP in RA remains controversial . Thus , we performed this meta-analysis to assess the efficacy and safety of CZP in the treatment of RA patients . METHODS : Eligible studies were randomized controlled trials ( RCTs ) that evaluated the efficacy and safe of CZP in the patients with active RA . The primary outcome was American College of Rheumatology 20 % ( ACR20 ) , and secondary outcome were ACR50 , ACR70 , disease activity , patient-reported outcomes ( PROs ) , and adverse events . A fixed-effect model or random-effect model was used to pool the estimates , depending on the absence or presence of heterogeneity among the included studies . RESULTS : Nine RCTs with a total of 5228 patients were included in this meta-analysis , and all of the patients were administered CZP or placebo . The pooled results showed that CZP significantly improved the ACR20 , ACR50 , ACR70 response rates , and physical function . CZP was associated with a statistically significant reduction in Disease Activity Score in 28 joints-Erythrocyte sedimentation rate , arthritis pain , and fatigue . Patients who received CZP treatment did not have a higher incidence of treatment-related adverse events , no matter in any intensity . CONCLUSIONS : CZP 200 or 400mg in the treatment of active RA significantly reduced the RA signs and symptoms , and improved physical function as compared with the placebo . More large-scale RCTs are needed to evaluate the long-term efficacy and safety of CZP in the treatment of active RA . 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 . Anti-allergic effects of nilotinib on mast cell-mediated anaphylaxis like reactions . DB04868 is a new orally bioavailable potent tyrosine kinase inhibitor that is used for the treatment of P11274 - P00519 -positive chronic myelogenous leukemia . However , its effect on mast cell-mediated anaphylactic reaction is still not known . The present study aimed to investigate the effect of nilotinib on the anaphylactic allergic reaction and study its possible mechanism(s) of action . DB04868 administration prevented systemic anaphylaxis in mice , mediated by compound 48/80 , in a dose- and time-dependent manner . Also , nilotinib significantly inhibited ( P < 0.05 ) allergic paw edema in rats . Furthermore , nilotinib significantly decreased ( P < 0.05 ) the IgE-mediated passive cutaneous anaphylaxis in a dose dependent manner . In addition , nilotinib dose-dependently reduced histamine release from the rat peritoneal mast cells activated either by compound 48/80 or by ovalbumin . Moreover , nilotinib attenuated the secretion of pro-inflammatory cytokine , tumor necrosis factor ( P01375 ) -α expression in the rat peritoneal mast cells . These findings provide evidence that nilotinib inhibits mast cell-derived immediate-type allergic reactions and so it could be a candidate as an anti-allergic agent . A fibronectin peptide redirects DB00102 / P09619 complexes to macropinocytosis-like internalization and augments DB00102 survival signals . Growth factor-binding domains identified in various extracellular matrix proteins have been shown to regulate growth factor activity in many ways . Recently , we identified a fibronectin peptide ( P12 ) that can bind platelet-derived growth factor BB ( DB00102 ) and promote adult human dermal fibroblast ( AHDF ) survival under stress . In vivo experiments in a porcine burn injury model showed that P12 limited burn injury progression , suggesting an active role in tissue survival . In this report , we explored the molecular mechanism of this peptide in ADHF under nutrient deprivation . Our results showed that P12 acted like some cell-penetrating peptides in that it redirected ligand-bound PDGF receptor ( P09619 ) from the clathrin-dependent endocytic pathway to a slower , macropinocytosis-like pathway . P12 slowed internalization and degradation of DB00102 , augmented its survival signals , and promoted cell survival after nutrient removal . Our findings demonstrate a mechanism for a potential therapeutic peptide that increases cell and tissue survival by acting as a cofactor to DB00102 . Light and X-ray scattering show decorin to be a dimer in solution . P07585 is a widely distributed member of the extracellular matrix small leucine-rich repeat glycoprotein/proteoglycan family . For investigation of its physical properties , decorin from two sources ( young steer skin and a recombinant adenovirus ) was used . The first sample was extracted into 7 m urea and purified , while the second was isolated from medium conditioned by 293A cells infected with adenovirus and purified without chaotropes . The only chemical differences detected between these materials were a slightly shorter glycosaminoglycan chain and the retention of the propeptide on the latter . Circular dichroism spectra of the two samples were virtually identical , showing a high proportion of beta-sheet and beta-turn and little alpha-helix . The protein cores were completely denatured in 2.25 m guanidine HCl ( GdnHCl ) but recovered their secondary structure on removal of chaotrope . Light scattering of material eluted from gel-filtration columns in DB03754 -buffered saline , pH 7.0 , gave molecular mass values of 165 +/- 1 kDa and 84.6 +/- 4 kDa for intact decorin and the glycoprotein core produced by digestion with chondroitin ABC lyase , respectively . Intact recombinant prodecorin had a mass of 148 +/- 18 kDa . These values , which are double those estimated from SDS gel electrophoresis or from the known sequences and compositions , were halved in 2.5 m GdnHCl . Data from solution x-ray scattering of intact decorin and its core in DB03754 -buffered saline are consistent with a dimeric particle whose protein component has a radius of gyration of 31.6 +/- 0.4 A , a maximum diameter of 98 +/- 5 A , and approximates two intertwined C shapes . DB06403 . Elevated endothelin ( ET ) -1 levels are strongly correlated with the pathogenesis and prognosis of pulmonary arterial hypertension ( PAH ) . DB06403 is an orally active , highly selective P25101 receptor antagonist with > 4000-fold higher selectivity over the ETB receptor . In two large , well designed , 12-week , placebo-controlled , phase III trials ( ARIES-1 , n = 202 and ARIES-2 , n = 192 ) in patients with PAH ( WHO group I ) , ambrisentan 2.5-10 mg once daily significantly increased 6-minute walk distance by 31-59 m from baseline ( primary outcome measure ) versus placebo . The incidence of clinical worsening ( secondary outcome measure ) was significantly delayed for the combined ambrisentan 5 mg once daily groups versus the combined placebo groups from ARIES-1 and -2 . At week 12 , WHO functional class distribution was significantly improved with once-daily ambrisentan 5 mg , and Borg dyspnoea scores were significantly improved with ambrisentan 2.5-10 mg versus placebo in combined data from the ARIES-1 and -2 trials . The beneficial effects of ambrisentan on exercise capacity , WHO functional class and Borg dyspnoea scores seen at 12 weeks were maintained at 48 weeks in the ARIES-E phase III extension trial ( n = 361 ) . One-year survival rates with ambrisentan were 95-97 % . Treatment with ambrisentan for up to 2.8 years was generally well tolerated in clinical trials . 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 . Leukotriene D4-induced increases in cytosolic calcium in THP-1 cells : dependence on extracellular calcium and inhibition with selective leukotriene D4 receptor antagonists . Agonist-induced changes in intracellular calcium ion concentration ( [Ca++]i ) were examined in human monocytic leukemia THP-1 cells loaded with fura 2/acetoxymethyl ester ( fura 2/AM ) . Leukotriene (LT)D4 induced a concentration-dependent biphasic response consisting of a transient phase ( up to 5-fold peak increase ) followed by a sustained phase , showing characteristics of a receptor-operated calcium channel . Homologous desensitization to LTD4 was observed . The responses to LTD4 were reduced by 80 to 90 % in calcium-free buffer . The responses to LTD4 in a calcium-free buffer were dependent upon the duration of prior exposure of the cells to a calcium-free environment . The response at 30 or 60 min after exposure to calcium-free buffer was greater than that at earlier time points ( time-dependent sensitization ) . Similar responses were obtained with THP-1 cells exposed to DB00974 -containing buffer . It is speculated that such time-dependent sensitization is a result of changes at the receptor level . The responses to LTD4 were blocked by two specific LTD4 antagonists , MK-0571 and DB00549 , in a concentration-dependent manner . When given after addition of LTD4 , MK-0571 or DB00549 reversed the sustained phase of the LTD4-induced response , suggesting that maintenance of the response requires persistent activation of the Q9Y271 . DB00549 was 5 to 10 times more potent than MK-0571 ( IC50 values of 1.1 and 9.3 nM , respectively ) , in agreement with results from radioligand binding studies reported separately . IKK-dependent activation of NF-κB contributes to myeloid and lymphoid leukemogenesis by P11274 - P00519 . The product of the Ph chromosome , the P11274 - P00519 tyrosine kinase activates diverse signaling pathways in leukemic cells from patients with chronic myeloid leukemia ( CML ) and Ph(+) B-cell acute lymphoblastic leukemia ( B-ALL ) . Previous studies showed that nuclear factor κB ( NF-κB ) is activated in P11274 - P00519 -expressing cells , but the mechanism of activation and importance of NF-κB to the pathogenesis of P11274 - P00519 -positive myeloid and lymphoid leukemias are unknown . Coexpression of P11274 - P00519 and a superrepressor mutant of inhibitory NF-κB α ( IκBαSR ) blocked nuclear p65/RelA expression and inhibited the proliferation of Ba/ P13726 cells and primary P11274 - P00519 -transformed B lymphoblasts without affecting cell survival . In retroviral mouse models of CML and B-ALL , coexpression of IκBαSR attenuated leukemogenesis , prolonged survival , and reduced myeloid leukemic stem cells . Coexpression of dominant-negative mutants of IκB kinase α ( IKKα ) / O15111 or IKKβ/ O14920 also inhibited lymphoid and myeloid leukemogenesis by P11274 - P00519 . Blockade of NF-κB decreased expression of the NF-κB targets c-MYC and BCL-X and increased the sensitivity of P11274 - P00519 -transformed lymphoblasts to P00519 kinase inhibitors . These results demonstrate that NF-κB is activated through the canonical IKK pathway and plays distinct roles in the pathogenesis of myeloid and lymphoid leukemias induced by P11274 - P00519 , validating NF-κB and IKKs as targets for therapy of Ph(+) leukemias . [ 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 . 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 . 5-Lipoxygenase mediates O14788 -induced osteoclast formation via the cysteinyl leukotriene receptor 1 . 5-Lipoxygenase ( P09917 ) catalyzes the formation of two major groups of leukotrienes , leukotriene B4 and cysteinyl leukotrienes ( CysLTs ) , and it has been implicated as a promising drug target to treat various inflammatory diseases . However , its role in osteoclastogenesis has not been investigated . In this study , we used mouse bone marrow-derived macrophages ( BMMs ) to show that P09917 inhibitor suppresses O14788 -induced osteoclast formation . Inhibition of P09917 was associated with impaired activation of multiple signaling events downstream of Q9Y6Q6 , including P29323 and p38 phosphorylation , and IκB degradation , followed by a decrease in O95644 expression . Ectopic overexpression of a constitutively active form of O95644 partly rescued the antiosteoclastogenic effect of P09917 inhibitor . The knockdown of P09917 in BMMs also resulted in a significant reduction in O14788 -induced osteoclast formation , accompanied by decreased expression of O95644 . Similar effects were shown with CysLT receptor (CysLTR)1/2 antagonist and small RNA for Q9Y271 in BMMs , indicating the involvement of CysLT and Q9Y271 in P09917 -mediated osteoclastogenesis . Finally , P09917 inhibitor suppressed LPS-induced osteoclast formation and bone loss in the in vivo mouse experiments , suggesting a potential therapeutic strategy for treating diseases involving bone destruction . Taken together , the results of this study demonstrate that P09917 is a key mediator of O14788 -induced osteoclast formation and possibly a novel therapeutic target for bone-resorption diseases . [ Proteolytic processing by dipeptidyl aminopeptidase IV generates receptor selectivity for peptide YY ( P10082 ) ] . Two receptor subtypes , Q03519 and P28062 , are known to mediate P10082 biological activity . P10082 1-36 binds to Q03519 and P28062 receptors with equal affinity , whereas the second endogenous form of P10082 , DB05004 , selectively binds to P28062 receptors . Dipeptidyl cleavage thus transforms an unselective Y agonist into a highly selective P28062 agonist , DB05004 . The enzyme responsible for this processing is unknown . Since P10082 has a proline in the penultimate position it is protected from the attack of most unspecific exopeptidases . Only a few exopeptidases are theoretically capable of generating DB05004 from P10082 1-36 . Of the enzymes tested only the dipeptidyl aminopeptidase IV ( P27487 , E.C. 3.4.14.5 ) cleaved DB00135 -Pro from P10082 1-36 with high activity . Since P27487 is found on the endothelial surface and brush border membranes it can be considered a candidate enzyme for generating DB05004 in vivo , thereby regulating the ratio of Q03519 / P49146 stimulation by P10082 . 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 . DB08904 : a new biologic targeting rheumatoid arthritis . The past decade has been an exciting period for clinical research and patient care in rheumatoid arthritis . This is mostly due to targeted biologic agents that have changed the outcome of this disease . DB08904 ( Cimzia(®) , UCB Inc. , GA , USA ) , which targets P01375 -α with a different mechanism of action than widely used biologics , was initially investigated for Crohn 's disease but has now been shown to be effective for rheumatoid arthritis . There have been three significant clinical trials demonstrating the efficacy of certolizumab pegol in active rheumatoid arthritis ; two with combination methotrexate and one with monotherapy . This article will summarize the data from those trials and compare some of the characteristics of certolizumab pegol to conventional disease-modifying antirheumatic drugs and other biologic agents . Treatment recommendations are beyond the scope of this review ; however , with many options available , there will be annotations on current trends in the care of this chronic disease . DB00644 induction of extracellular-signal regulated kinase is blocked by inhibition of calmodulin . Our previous studies demonstrate that DB00644 -induced P29323 activation required influx of extracellular Ca2+ in alphaT3-1 and rat pituitary cells . In the present studies , we examined the hypothesis that calmodulin ( Cam ) plays a fundamental role in mediating the effects of Ca2+ on P29323 activation . Cam inhibition using W7 was sufficient to block DB00644 -induced reporter gene activity for the c-Fos , murine glycoprotein hormone alpha-subunit , and MAPK phosphatase ( MKP ) -2 promoters , all shown to require P29323 activation . Inhibition of Cam ( using a dominant negative ) was sufficient to block DB00644 -induced P29323 but not c-Jun N-terminal kinase activity activation . The Cam-dependent protein kinase ( CamK ) II inhibitor KN62 did not recapitulate these findings . DB00644 -induced phosphorylation of Q02750 and c-Raf kinase was blocked by Cam inhibition , whereas activity of phospholipase C was unaffected , suggesting that Ca2+/Cam modulation of the P29323 cascade potentially at the level of c-Raf kinase . Enrichment of Cam-interacting proteins using a Cam agarose column revealed that c-Raf kinase forms a complex with Cam . Reconstitution studies reveal that recombinant c-Raf kinase can associate directly with Cam in a Ca2+-dependent manner and this interaction is reduced in vitro by addition of W7 . Cam was localized in lipid rafts consistent with the formation of a Ca2+-sensitive signaling platform including the P30968 and c-Raf kinase . These data support the conclusion that Cam may have a critical role as a Ca2+ sensor in specifically linking Ca2+ flux with P29323 activation within the DB00644 signaling pathway . Inhibition of canine exocrine pancreatic secretion by peptide YY is mediated by P10082 -preferring P28062 receptors . It is still unclear , which receptor subtype , Q03519 and/or P28062 , mediates the inhibitory action of P10082 on exocrine pancreatic secretion . The present study was undertaken to characterize functionally the Y receptor subtype that mediates the inhibition of exocrine pancreatic secretion by peptide YY ( P10082 ) . In eight conscious dogs with chronic gastric and pancreatic fistulas , we compared the action of intravenous infusion of 200 and 400 pmol/kg/h of the Y receptor agonists P10082 1-36 , DB05004 , P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 on the pancreatic secretory response to secretin ( 20.5 pmol/kg/h ) and cerulein ( 29.6 pmol/kg/h ) . P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 were also studied by giving a fivefold dose ( 1,000 and 2,000 pmol/kg/h ) . P10082 1-36 and the P49146 agonist DB05004 significantly inhibited pancreatic secretory responses to secretin and cerulein , whereas inhibition by P01303 1-36 and the P49146 agonist P10082 13-36 was attainable only at doses of 1,000 and 2,000 pmol/kg/h . The Q03519 receptor agonist Pro34PYY 1-36 was without effect on pancreatic secretion . We conclude that in dogs the inhibition of exocrine pancreatic secretion by P10082 is mediated via P28062 receptors of a P10082 -preferring subtype . Antineoplastic mechanisms of niclosamide in acute myelogenous leukemia stem cells : inactivation of the NF-kappaB pathway and generation of reactive oxygen species . NF-kappaB may be a potential therapeutic target for acute myelogenous leukemia ( AML ) because NF-kappaB activation is found in primitive human AML blast cells . In this report , we initially discovered that the potent antineoplastic effect of niclosamide , a Food and Drug Administration-approved antihelminthic agent , was through inhibition of the NF-kappaB pathway in AML cells . DB06803 inhibited the transcription and DNA binding of NF-kappaB . It blocked tumor necrosis factor-induced P25963 phosphorylation , translocation of p65 , and expression of NF-kappaB-regulated genes . DB06803 inhibited the steps TAK1 --> O15111 ( IKK ) and IKK --> P25963 . DB06803 also increased the levels of reactive oxygen species ( ROS ) in AML cells . Quenching ROS by the glutathione precursor DB06151 attenuated niclosamide-induced apoptosis . Our results together suggest that niclosamide inhibited the NF-kappaB pathway and increased ROS levels to induce apoptosis in AML cells . On translational study of the efficacy of niclosamide against AML , niclosamide killed progenitor/stem cells from AML patients but spared those from normal bone marrow . DB06803 was synergistic with the frontline chemotherapeutic agents cytarabine , etoposide , and daunorubicin . It potently inhibited the growth of AML cells in vitro and in nude mice . Our results support further investigation of niclosamide in clinical trials of AML patients . Pharmacological investigation of the role of leukotrienes in the pathogenesis of experimental NSAID gastropathy . The role of leukotrienes in the pathogenesis of acute gastric ulceration induced by nonsteroidal antiinflammatory drugs was investigated using a rat model . One part of the study involved oral pretreatment with a leukotriene synthesis inhibitor 1 h prior to administration of indomethacin ( 20 mg/kg per os ) . Three hours after indomethacin , the extent of macroscopically visible gastric damage was determined , and gastric LTB4 synthesis was determined . The compounds tested were PF-5901 , A-64077 , nordihydroguaiaretic acid , and L-698,037 . Each compound produced dose-related inhibition of gastric LTB4 synthesis and a parallel reduction in the severity of indomethacin-induced damage . The antioxidant properties of these compounds was assessed using an in vitro assay . There was no correlation between the antioxidant properties of the compounds and their ability to reduce the severity of indomethacin-induced gastric damage . In the second part of the study , the effects of intravenous , administration of LTD4 and LTB4 receptor antagonists on indomethacin-induced gastric epithelial damage ( measured by permeability to [51Cr] DB00974 ) were assessed . The two Q9Y271 antagonists ( MK-571 and DB00549 ) significantly reduced the permeability changes induced by indomethacin , while the two LTB4 antagonists ( SC-41930 and LY-255,283 ) were without significant effect . Despite the reduction of gastric epithelial injury , blockade of LTD4 receptors did not markedly affect the extent of macroscopically visible injury . These data are consistent with the hypothesis that leukotrienes contribute to the epithelial injury and macroscopically visible damage induced by NSAIDs . However , it remains unclear to what extent leukotrienes are involved in the initiation of the injury , as opposed to its amplification . 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 . Safety and efficacy evaluation of ambrisentan in pulmonary hypertension . INTRODUCTION : Pulmonary arterial hypertension ( PAH ) is characterized by an increase in pulmonary vascular resistance , which can lead to right heart failure and death . P05305 binding P25101 and ETB receptors seem to play a critical role in the pathogenesis and progression of the disease , and oral endothelin receptor antagonists ( ERAs ) have been shown to be an effective treatment . DB00559 and P25101 -selective ambrisentan are the ERAs currently available for PAH treatment . AREAS COVERED : On the basis of the analysis of the literature , this paper addresses the efficacy and safety of ambrisentan in the treatment for PAH . EXPERT OPINION : DB06403 has shown an efficacy comparable with other ERAs . Compared with DB00559 , ambrisentan seems to have a better safety profile with regards to hepatic safety and drug-drug interactions . On the other hand , ambrisentan shows a higher rate of other adverse events , such as nasal congestion and peripheral edema . DB06403 is a viable option for PAH treatment . However , there is still a need for more robust data about long-term mortality , treatment in non-PAH pulmonary hypertension ( PH ) ( such as PH due to left heart disease and PH due to chronic hypoxic lung diseases ) and combination therapy . 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 . 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 % . Synthesis and biological activity of DB00644 antagonists modified at position 3 with 3-(2-methoxy-5-pyridyl)-alanine . DB06699 is a potent very long-acting DB00644 antagonist after subcutaneous administration . In this paper , we describe the synthesis of two analogs of degarelix incorporating racemic 3-(2-methoxy-5-pyridyl)-alanine ( 2-OMe-5Pal , 5 ) at position 3 . The two diastereomers were separated by reverse-phase high-performance liquid chromatography ( RP-HPLC ) and the absolute stereochemistry at position 3 in the peptides was determined by enzymatic digestion with proteinase K . These analogs were tested in vitro for their ability to antagonize the P30968 and in vivo for duration of action in a castrated male rat assay . Analog 7 with D2-OMe-5Pal was potent in vitro ( IC50 = 5.22 nM ) ; however , analog 8 with Q401N2 -OMe-5Pal at position 3 in degarelix lost potency as an antagonist of the human P30968 ( IC50 = 36.95 nM ) . Both the analogs were found to be short-acting in vivo . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . 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 . Sequence motifs in IL-4R alpha mediating cell-cycle progression of primary lymphocytes . P05112 signaling through the IL-4Ralpha chain regulates the development and proliferation of the Th2 lineage of effector P01730 (+) T cells . Analyses of the IL-4R in factor-dependent cell lines led to the development of two apparently conflicting models of the primary structural determinants of IL-4R-mediated proliferative signaling . In one model , proliferation was dependent on the first conserved tyrosine in the cytoplasmic tail ( Q03519 ) , while in the second , proliferation was independent of cytoplasmic tyrosines . We found that in activated primary T cells , mutation of only the Q03519 residue resulted in a modest decrease in P05112 -induced S phase entry , a further decrease in cell-cycle completion , and a complete failure of P05112 to induce p70S6 kinase phosphorylation . Consistent with a role for the PI3K/mammalian target of rapamycin pathway in mediating cytokine acceleration of G(2)/M transit , pretreatment of activated T cells with rapamycin resulted in only a modest decrease in P05112 -induced S phase entry , but a total block of cell-cycle completion . Strikingly , IL-4Ralpha chains that lacked all cytoplasmic tyrosines were competent to signal for P42229 phosphorylation , mediated efficient S phase entry , and promoted cell-cycle progression . The ability of tyrosine-deficient IL-4Rs to mediate proliferative signaling and P35610 phosphorylation was absolutely dependent on the presence of an intact ID-1 region . These findings show that IL-4Ralpha lacking cytoplasmic tyrosine residues is competent to induce ID-1-dependent proliferation , and indicate that P05112 can promote G(2)/M progression via activation of the mammalian target of rapamycin pathway initiated at the Q03519 residue . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment .

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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 . 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 . Metronomic chemotherapy dosing-schedules with estramustine and temozolomide act synergistically with anti- P35968 antibody to cause inhibition of human umbilical venous endothelial cell growth . The effects of ' metronomic ' or extended chemotherapy dosing schedules ( ECS ) are mediated through poorly understood anti-angiogenic mechanisms . ECS combined with biological anti-angiogenic agents have produced promising pre-clinical results . MATERIALS AND METHODS : We have expanded the list of agents with an in vitro ECS profile to include the methylating agent temozolomide ( DB00853 ) and the anti-mitotic agent estramustine ( Estracyt ) . These agents were also combined with a specific anti-angiogenic inhibitor DB06101 and a non-specific agent with anti-angiogenic properties , Compound 5h . The in vitro HUVEC ECS model system was optimised and cell proliferation assays undertaken . RESULTS : As a single agent , estramustine inhibited endothelial cell proliferation with an IC50 of 4.5 microM and was active at 10-33 % of the maximum tolerated dose ( MTD ) from clinical schedules , whilst temozolomide had IC50 of 6.6 microM and was active at 1-6 % of MTD . In combination , significant synergy was seen with DB06101 in combination with either drug , whilst modest additive effects were observed with Compound 5h . None of the combinations resulted in significant cytotoxicity or apoptosis . DISCUSSION : The results show that ECS of temozolomide and estramustine can be significantly enhanced when combined with specific anti-angiogenic inhibitors in an in vitro HUVEC system . 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 . Effects of matrine on proliferation and apoptosis of cultured retinoblastoma cells . BACKGROUND : Extracted from the traditional Chinese medicine of Kushen , matrine is an alkaloid with potential anti-neoplastic and anti-inflammatory effects . Here , we examined the effect of matrine on proliferation and apoptosis of cultured retinoblastoma cells . METHODS : The retinoblastoma cell lines Y79 , WERI- P06400 and SO-RB50 were treated with matrine in increasing concentrations from 0. P35326 .1 mg/ml for 24 hours , and the cell proliferation rate was measured . The cells were exposed to matrine at 50 % inhibition concentration ( IC50 ) for 12 , 24 and 48 hours . Cell cycle was analyzed by flow cytometry , concentration of proteins regulating cell cycle and apoptosis was determined by Western blot , apoptosis rate was measured by TUNEL staining , and cell morphology was assessed by electron transmission microscopy . RESULTS : The retinoblastoma cell lines Y79 , WERI- P06400 and SO-RB50 showed an increased inhibition of cell proliferation with increasing matrine concentrations . Applying the IC50 concentration of matrine , the alteration of the cell cycle , including a reduced percentage of the S phase , was significantly ( P < 0.01 ) associated with a longer treatment time by matrine . Correspondingly , the cell-cycle-associated proteins P21 and P27 were up-regulated and the protein cyclinD1 was down-regulated . The apoptosis-associated protein Bcl-2 was down-regulated , and Bax was up-regulated . In a similar manner , the apoptosis rate was significantly increased with longer treatment time . CONCLUSIONS : Matrine added to cultures of immortalized retinoblastoma cells led to a reduced tumor cell proliferation , decreased rate of mitosis and an increased tumor cell apoptosis , paralleled by corresponding changes in the proteins regulating the cell cycle or apoptosis . P10275 mediates non-genomic activation of phosphatidylinositol 3-OH kinase in androgen-sensitive epithelial cells . Androgens are known to modulate many cellular processes such as cell growth and survival by binding to the androgen receptor ( AR ) and activating the transcription of target genes . Recent data suggested that AR can also mediate non-transcriptional actions outside the nucleus in addition to its ligand-inducible transcription factor function . Here , we describe a transcription-independent activation of the phosphatidylinositol 3-OH kinase ( P19957 -K ) signaling pathway by androgens . Using non-transformed androgen-sensitive epithelial cells , we show that androgens enhance the P19957 -K activity by promoting accumulation of phosphoinositide-3-P phospholipids in vitro . This activation is found in conjunction with an increased time-dependent phosphorylation of the downstream kinase AKT/protein kinase B on both DB00133 (473) and DB00156 (308) residues . Hormone-stimulated phosphorylation of AKT requires AR since incubation with the anti-androgen bicalutamide completely abolishes the androgen-stimulated AKT phosphorylation . Accordingly , we show that androgens increase AKT phosphorylation level in prostatic carcinoma PC3 cells only once they have been transfected with AR . Downstream , androgens enhance phosphorylation of transcription factor Q12778 ( Q12778 ) - Q9NUQ9 and proapoptotic Bad protein and promote cell survival as they can counteract an apoptotic process . We also report that non-genomic effects of androgens are based on direct interaction between AR and the p85alpha regulatory subunit of class I(A) P19957 -K . Together , these novel findings point out an important and physiologically relevant link between androgens and the P19957 -K/AKT signaling pathway in governing cell survival . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . 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 . 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 . 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 . Endothelial production of P23582 and its marked augmentation by transforming growth factor-beta . Possible existence of " vascular natriuretic peptide system " . P23582 ( P09543 ) , the third member of the natriuretic peptide family , is thus far known to be distributed mainly in the central nervous system and is considered to act as a neuropeptide , in contrast to atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) , which act as cardiac hormones . Recently , we and others have demonstrated that the P20594 receptor , which is selectively activated by P09543 , is localized not only in the central nervous system but in peripheral tissues , including blood vessels . This finding has made us speculate regarding the peripheral production of P09543 . In the present study , cultured endothelial cells were examined for P09543 production by RIA and Northern blot analysis . P09543 -like immunoreactivity was detected in the conditioned media of endothelial cells . Northern blot analysis detected CNPmRNA with a size of 1.2 kb . In addition , transforming growth factor ( TGF ) -beta , one of the key growth factors for vascular remodeling , markedly stimulated the expression of CNPmRNA and induced a tremendous increase in P09543 secretion . We could also detect P09543 transcript in the bovine thoracic aorta using the reverse transcription-polymerase chain reaction method . The present study demonstrates the endothelial production of P09543 and suggests that a member of the natriuretic peptide family may act as a local regulator in vascular walls . Since evidence for the pathophysiological importance of the vascular renin-angiotensin system has been accumulating and the natriuretic peptide system is known to be antagonistic to the renin-angiotensin system , the possible existence of " vascular natriuretic peptide system " may prove to be of physiological and clinical relevance . P10275 regulation of P55008 cyclin and cyclin-dependent kinase function in the CWR22 human prostate cancer xenograft . Human prostate cancer is initially dependent on androgens for growth , and androgen-dependent cells undergo apoptosis after castration . However , a subset of androgen-responsive cells survives and eventually proliferates in the absence of testicular androgen . The high levels of androgen receptor in both androgen-dependent and recurrent tumors led us to investigate androgen regulation of cell cycle proteins in human prostate cancer using the CWR22 xenograft . Cellular proliferation decreased dramatically in CWR22 tumors after castration . DB00624 propionate ( TP ) treatment of castrated mice restored cellular proliferation after 24-48 hours . Growth of CWR22 tumors in the absence of testicular androgen recurred several months after castration . P06493 and P24941 , and cyclin A and cyclin B1 messenger RNAs were decreased 6 days after castration , increased 6-12 hours after TP treatment , and were expressed at high levels in recurrent CWR22 tumors . Coimmunoprecipitated cyclin B1/ P06493 and cyclin D1/ P11802 protein complexes decreased after castration and increased after TP treatment of castrated mice . In addition , P06493 and P24941 kinase activities were upregulated by androgen in parallel with hyperphosphorylation of retinoblastoma ( Rb ) protein . Despite the absence of testicular androgen in recurrent CWR22 , the levels of these androgen-regulated cyclin/ CDK protein complexes and hyperphosphorylation of Rb were equal to or greater than in tumors from intact mice . The results indicate that androgen receptor regulates cellular proliferation by control of CDK and cyclins at the transcriptional level and by post-translational modifications that influence cell cycle protein activity . 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 . Comparative actions of insulin sensitizers on ion channels in vascular smooth muscle . Thiazolidinedione and isoxazolidinedione insulin sensitizers activate peroxisome proliferator-activated receptor gamma ( Q07869 gamma ) . Some thiazolidinediones modify ion channels in smooth muscles ; however , the mechanism by which their actions occur has not been clarified . We , thus , examined the effects of three thiazolidinediones ( troglitazone , pioglitazone , and rosiglitazone ) and isoxazolidinedione ( DB04971 ) , as well as an intrinsic ligand for Q07869 gamma , 15-deoxy-Delta(12,14) prostaglandin J(2) ( prostaglandin J(2) ) , on voltage-operated Ca(2+) currents ( I(Ca) ) , voltage-dependent K(+) currents ( I(Kv) ) , and Ca(2+)-activated K(+) currents ( I(Kca) ) , to clarify whether a thiazolidinedione structure or Q07869 gamma activation is related to their actions on ion channels . The whole-cell patch clamp method was used to record currents in smooth muscle cells from guinea-pig mesenteric arteries . Thiazolidinediones inhibited I(Ca) in a dose-dependent manner ( troglitazone > pioglitazone=rosiglitazone ) . DB00197 ( > or =1 microM ) and rosiglitazone ( 100 microM ) , but not pioglitazone , inhibited I(Kv) . Rosiglitazone ( > or =10 microM ) enhanced , troglitazone ( > or =1 microM ) inhibited , and pioglitazone did not affect I(Kca) . A high concentration of DB04971 ( 100 microM ) inhibited I(Ca) , I(Kv) , and I(Kca) to a similar extent . Prostaglandin J(2) enhanced I(Kca) , but affected neither I(Ca) nor I(Kv) . In summary , the three thiazolidinediones and isoxazolidinedione act differently on Ca(2+) and K(+) channels in vascular smooth muscle . The action of thiazolidinediones on I(Ca) could be attributed to specific regions of the molecules and not to activation of Q07869 gamma . Involvement of Q07869 gamma activation in the stimulation of I(Kca) is possible but should be tested further . Inhibition of ROS-activated p38MAPK pathway is involved in the protective effect of H2S against chemical hypoxia-induced inflammation in PC12 cells . We have demonstrated the neuroprotection of hydrogen sulfide ( H2S ) against chemical hypoxia-induced injury by inhibiting p38MAPK pathway . The present study attempts to evaluate the effect of H2S on chemical hypoxia-induced inflammation responses and its mechanisms in PC12 cells . We found that treatment of PC12 cells with cobalt chloride ( CoCl2 , a hypoxia mimetic agent ) enhanced P05231 secretion , nitric oxide ( NO ) generation and expression levels of inducible nitric oxide synthase ( P35228 ) and neuronal nitric oxide synthase ( P29475 ) . L-canavanine , a selective P35228 inhibitor , partly blocked CoCl2-induced cytotoxicity , apoptosis and mitochondrial insult . In addition , DB02207 ( 7-NI ) , an inhibitor of P29475 , also partly attenuated the CoCl2-induced cytotoxicity . The inhibition of p38MAPK by SB203580 ( a selective p38MAPK inhibitor ) or genetic silencing of p38MAPK by RNAi ( Si-p38 ) depressed not only CoCl2-induced P35228 expression , NO production , but also P05231 secretion . In addition , N-acetyl-L-cysteine , a reactive oxygen species ( ROS ) scavenger , conferred a similar protective effect of SB203580 or Si-p38 against CoCl2-induced inflammatory responses . Importantly , pretreatment of PC12 cells with exogenous application of sodium hydrosulfide ( a H2S donor , 400 μmol/l ) for 30 min before exposure to CoCl2 markedly attenuated chemical hypoxia-stimulated P35228 and P29475 expression , NO generation and P05231 secretion as well as p38MAPK phosphorylation in PC12 cells . Taken together , we demonstrated that p38MAPK- P35228 pathway contributes to chemical hypoxia-induced inflammation and that H2S produces an anti-inflammatory effect in chemical hypoxia-stimulated PC12 cells , which may be partly due to inhibition of ROS-activated p38MAPK- P35228 pathway . Rapid inhibition of vasoconstriction in renal afferent arterioles by aldosterone . DB04630 has been suggested to elicit vessel contraction via a nongenomic mechanism . We tested this proposal in microdissected , perfused rabbit renal afferent arterioles . DB04630 had no effect on internal diameter in concentrations from 10(-10) to 10(-5) mol/L , but aldosterone abolished the ability of 100 mmol/L DB00761 to induce vascular contraction . The inhibitory effect of aldosterone was observed from 1 pmol/L . The inhibitory effect was significant after 5 minutes and maximal after 20 minutes and was fully reversible . DB00970 ( 10(-6) mol/L ) prolonged the effect of aldosterone . The effect was abolished by the mineralocorticoid receptor antagonist spironolactone ( 10(-7) mol/L ) but not by the glucocorticoid receptor antagonist mifepristone ( 10(-6) mol/L ) . The K+-mediated increase of intracellular calcium concentration in afferent arterioles was not affected by aldosterone . P08235 was detected by reverse transcription-polymerase chain reaction and immunohistochemistry in rat renal vasculature and rabbit endothelial cells . Inhibition of phosphatidylinositol ( PI ) -3 kinase with LY 294002 ( 3x10(-6) mol/L ) restored sensitivity to K+ in the presence of aldosterone , and afferent arterioles were immunopositive for P19957 kinase subunit P42336 . Inhibition of NO formation by L-NAME ( 10(-4) mol/L ) or inhibition of soluble guanylyl cyclase with 1H-(1,2,4)Oxadiazolo[4,3-a]quinoxaline-1-one restored K+-induced vasoreactivity in the presence of aldosterone . Similar to aldosterone , the NO donor sodium nitroprusside inhibited K+-induced vascular contraction . DB02424 ( 10(-6) mol/L ) , an inhibitor of heat shock protein 90 , abolished aldosterone-induced vasorelaxation . We conclude that aldosterone inhibits depolarization-induced vasoconstriction in renal afferent arterioles by a rapid nongenomic mechanism that is initiated by mineralocorticoid receptor activation and involves P19957 kinase , protein kinase B , and heat shock protein 90-mediated stimulation of NO generation . Synthetic cyclin dependent kinase inhibitors . New generation of potent anti-cancer drugs . The unsatisfactory results of current anti-cancer therapies require the active search for new drugs , new treatment strategies and a deeper understanding of the host-tumour relationship . From this point of view , the drugs with a capacity to substitute the functions of altered tumour suppressor genes are of prominent interest . Since one of the main functions of oncosuppressors is to mediate cell cycle arrest via modification of cyclin dependent kinases ( CDKs ) activity , the compounds with ability to substitute altered functions of these genes in neoplastic cells are of prominent interest . Synthetic inhibitors of cyclin dependent kinases ( CDKIs ) are typical representatives of such drugs . DB02116 ( OC ) , flavopiridol ( FP ) , DB04699 I ( BL ) and their derivatives selectively inhibit CDKs and thus constrain tumor cell proliferation under in vitro and/or in vivo conditions . We originally discovered OC and its inhibitory activity toward P06493 family of CDKs , and recently reported the induction of apoptosis and tumor regression following OC application . Moreover , the OC family of synthetic CDKIs has the capacity of directly inhibit P50613 , the principal enzyme required for activating other CDKs , and thus these compounds are the first known P50613 inhibitors . Its unique mechanism of action and potent anti-cancer activity under both in vitro and in vivo conditions provide a unique tool to inhibit tumour cell proliferation , and to selectively induce apoptosis in neoplastic tissues . The mechanisms of anti-cancer activities of FP , BL , OC and related synthetic CDKIs are compared and discussed in this paper . 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 . Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway . DB00624 supplementation increases skeletal muscle mass and decreases fat mass ; however , the underlying mechanisms are unknown . We hypothesized that testosterone regulates body composition by promoting the commitment of mesenchymal pluripotent cells into myogenic lineage and inhibiting their differentiation into adipogenic lineage . Mouse C3H 10T1/2 pluripotent cells were treated with testosterone ( 0-300 nM ) or dihydrotestosterone ( DB02901 , 0-30 nM ) for 0-14 d , and myogenic conversion was evaluated by immunocytochemical staining for early ( MyoD ) and late ( myosin heavy chain II ; MHC ) myogenic markers and by measurements of MyoD and MHC mRNA and protein . Adipogenic differentiation was assessed by adipocyte counting and by measurements of peroxisomal proliferator-activated receptor gamma 2 ( Q07869 gamma 2 ) mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . The number of MyoD+ myogenic cells and MHC+ myotubes and MyoD and MHC mRNA and protein levels increased dose dependently in response to testosterone and DB02901 treatment . Both testosterone and DB02901 decreased the number of adipocytes and down-regulated the expression of Q07869 gamma 2 mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . P10275 mRNA and protein levels were low at baseline but increased after testosterone or DB02901 treatment . The effects of testosterone and DB02901 on myogenesis and adipogenesis were blocked by bicalutamide . Therefore , testosterone and DB02901 regulate lineage determination in mesenchymal pluripotent cells by promoting their commitment to the myogenic lineage and inhibiting their differentiation into the adipogenic lineage through an androgen receptor-mediated pathway . The observation that differentiation of pluripotent cells is androgen dependent provides a unifying explanation for the reciprocal effects of androgens on muscle and fat mass in men .

MH_train_1553

MH_train_1553

MH_train_1553

interacts_with DB00502?

multiple_choice

Modular competition driven by DB01221 receptor subtypes in spike-timing-dependent plasticity . N-methyl-d-aspartate receptors ( NMDARs ) play a critical role in transducing neuronal activity patterns into changes in synaptic strength . However , how they mediate this transduction in response to physiological stimuli has remained elusive . In particular , it has been debated whether different NMDAR subtypes play opposing signaling roles in synaptic plasticity . Using perforated patch-clamp recordings from pairs of synaptically connected glutamatergic neurons in dissociated hippocampal culture , we found that spike-timing-dependent potentiation induced by pairing pre- and postsynaptic spikes required the activation of a fast component of NMDAR current that is likely to be mediated by Q12879 -containing NMDARs ( Q12879 -NRs ) . In contrast , spike-timing-dependent depression required a slow component of NMDAR current carried by Q13224 -containing NMDARs ( Q13224 -NRs ) . CV analysis showed that the locus of this depression was primarily presynaptic in pairs of cells making strong synaptic connections , whereas weaker synapses showed no clear preference for pre- or postsynaptic expression . This depression was not significantly reduced by antagonism of the P21554 receptor , in contrast to spike-timing-dependent depression in the neocortex that requires presynaptic P21554 signaling . With blockade of Q13224 -NRs , spike triplets that contained both potentiating and depressing spike-timing components induced net potentiation . However , when the putative Q12879 -NR population is inhibited , these spike triplets resulted in either depression or no net change , depending on the temporal order of the spike-timing components . These results imply a dynamic competition between signaling modules that can be biased by differentially antagonizing NMDAR subtypes during the induction of spike-timing-dependent plasticity . Using a simple model , we show that such a modular competition recapitulates our observations . 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 . Novel 3,4-diarylpyrazolines as potent cannabinoid P21554 receptor antagonists with lower lipophilicity . Novel 3,4-diarylpyrazolines 1 as potent P21554 receptor antagonists with lipophilicity lower than that of DB05077 are described . The key change is the replacement of the arylsulfonyl group in the original series by a dialkylaminosulfonyl moiety . The absolute configuration ( 4S ) of eutomer 24 was established by X-ray diffraction analysis and 24 showed a close molecular fit with rimonabant in a P21554 receptor-based model . Compound 17 exhibited the highest P21554 receptor affinity ( Ki = 24 nM ) in this series , as well as very potent P21554 antagonistic activity ( pA2 = 8.8 ) and a high P21554 /CB2 subtype selectivity ( approximately 147-fold ) . Regulation of the adrenal androgen biosynthesis . The human adrenal reticularis produces the so-called adrenal androgens , dehydroepiandrosterone ( DB01708 ) and DB01708 -sulfate ( DB01708 -S ) . As opposed to the cortisol and aldosterone little is known regarding the mechanisms that regulate the production of the adrenal androgens . Several recent studies have shown that type II 3beta-hydroxysteroid dehydrogenase ( P26439 ) , cytochrome b5 ( P00167 ) , and steroid sulfotransferase ( Q06520 ) play an important role in the regulation of adrenal androgen production . Specifically , adrenal production of DB01708 -S is correlated with reticularis expression of Q06520 and P00167 . In contrast , P26439 has an inverse correlation with adrenal androgen production likely due to its unique ability to remove precursors from the pathway leading to DB01708 . Therefore , its expression is limited to the adrenal glomerulosa/fasciculata but not in reticularis . The differential expression of these three proteins appears to be critical for reticularis function . In this review , we focus on studies that have begun to define the mechanisms regulating the transcription of these genes . Understanding the mechanisms controlling differential expression of these proteins should provide novel information about the human adrenal reticularis and its production of DB01708 and DB01708 -S . 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 . 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 . Physical activity correlates with glutamate receptor gene expression in spinally-projecting RVLM neurons : a laser capture microdissection study . Physical inactivity is an important risk factor in the development of cardiovascular disease . The rostral ventrolateral portion of the medulla ( RVLM ) is composed of heterogeneous populations of neurons that are involved in the regulation of the cardiovascular system . Because of functional heterogeneity , studying the changes in the gene expression of this specific population of neurons within the RVLM is challenging . In the present study , a fluorescent retrograde tracer was injected into the spinal cord to specifically label bulbospinal RVLM neurons in sedentary and active rats . Laser capture microdissection ( DB01627 ) was then employed to collect the fluorescently labeled neurons from sections encompassing the rostrocaudal extent of the RVLM . RNA extracted from the neurons was used in qRT-PCR analysis . Changes in gene expression levels of glutamate and GABA receptor subunits were compared between sedentary and physically active rats . P42263 subunit showed a significant negative correlation between total running distance and its relative gene expression in active rats . There were no significant difference in the gene expression of DB01221 ( Q9UHB4 , Q12879 , Q13224 , Q14957 and O15399 ) , AMPA ( P42261 , P42262 and P42263 ) and GABAA ( GABAA1 and GABAA2 ) receptor subunits . Overall , the present study demonstrates the feasibility of utilizing DB01627 to investigate the gene expression changes in a specific population of neurons in the RVLM . Correlation studies suggest that physical activity could contribute to neuroplasticity in the RVLM . Status epilepticus triggers early and late alterations in brain-derived neurotrophic factor and DB01221 glutamate receptor Grin2b DNA methylation levels in the hippocampus . Status epilepticus ( SE ) triggers abnormal expression of genes in the hippocampus , such as glutamate receptor subunit epsilon-2 ( Grin2b/Nr2b ) and brain-derived neurotrophic factor ( Bdnf ) , that is thought to occur in temporal lobe epilepsy ( TLE ) . We examined the underlying DNA methylation mechanisms and investigated whether these mechanisms contribute to the expression of these gene targets in the epileptic hippocampus . Experimental TLE was provoked by kainic acid-induced SE . Bisulfite sequencing analysis revealed increased Grin2b/Nr2b and decreased Bdnf DNA methylation levels that corresponded to decreased Grin2b/Nr2b and increased Bdnf mRNA and protein expression in the epileptic hippocampus . Blockade of DNA methyltransferase ( P26358 ) activity with zebularine decreased global DNA methylation levels and reduced Grin2b/Nr2b , but not Bdnf , DNA methylation levels . Interestingly , we found that P26358 blockade further decreased Grin2b/Nr2b mRNA expression whereas Q13224 protein expression increased in the epileptic hippocampus , suggesting that a posttranscriptional mechanism may be involved . Using chromatin immunoprecipitation analysis we found that P26358 inhibition restored the decreases in AP2alpha transcription factor levels at the Grin2b/Nr2b promoter in the epileptic hippocampus . P26358 inhibition increased field excitatory postsynaptic potential in hippocampal slices isolated from epileptic rats . Electroencephalography ( EEG ) monitoring confirmed that P26358 inhibition did not significantly alter the disease course , but promoted the latency to seizure onset or SE . Thus , DNA methylation may be an early event triggered by SE that persists late into the epileptic hippocampus to contribute to gene expression changes in TLE . 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 . P00797 and prorenin have no direct effect on aldosterone synthesis in the human adrenocortical cell lines H295R and HAC15 . INTRODUCTION : Transgenic rats expressing the human (pro)renin receptor ( h(P)RR ) have elevated plasma aldosterone levels despite unaltered levels , in plasma and adrenal , of renin and angiotensin II . MATERIALS AND METHODS : To investigate whether renin/prorenin-(P)RR interaction underlies these elevated aldosterone levels , the effect of (pro)renin on steroidogenesis was compared with that of angiotensin II in two (P)RR-expressing human adrenocortical cell lines , H295R and HAC15 . Angiotensin II rapidly induced extracellular signal-regulated kinase ( P29323 ) phosphorylation and increased the expression of STAR , P08686 , P19099 , and P05093 at 6 and 24 hours , whereas the expression of P05108 and P26439 remained unaltered . Incubation with renin or prorenin at nanomolar concentrations had no effect on the expression of any of the steroidogenic enzymes tested , nor resulted in P29323 phosphorylation . Angiotensin II , but not renin or prorenin , induced aldosterone production . CONCLUSION : Although the (P)RR is present in adrenocortical cells , renin and prorenin do not elicit P29323 phosphorylation nor directly affect steroid production via this receptor at nanomolar concentrations . Thus , direct (pro)renin-(P)RR interaction is unlikely to contribute to the elevated aldosterone levels in human (P)RR transgenic rats . This conclusion also implies that the aldosterone rise that often occurs during prolonged renin-angiotensin system blockade is rather due to the angiotensin II ' escape ' during such blockade . P15692 activates Q13224 phosphorylation through Dab1 pathway . Vascular endothelial growth factor ( P15692 ) and reelin are two major signaling pathways involved in many neuronal functions including neurogenesis and neuronal migration . Both P15692 and reelin have been shown to regulate DB01221 type glutamate receptor ( NMDAR ) activity via independent mechanisms . However , it is not known whether the above signaling pathways influence each other on NMDAR regulation . We demonstrate that Disabled 1 ( Dab1 ) , a downstream signaling molecule of reelin pathway mediates P15692 -induced regulation of NMDAR subunit Q13224 . Furthermore , P15692 treatment led to the association of P15692 receptor-2 ( Flk1 ) and reelin receptor ( apolipoprotein E receptor 2 , ApoER2 ) , and Dab1 as well as Q13224 activation were Flk1-dependent . Moreover , P15692 treatment could significantly rescue the deficits in phospho-Dab1 levels in reeler ( Reln-/- ) neurons . Our results suggest a major role of P15692 in the regulation of reelin signaling , and Dab1 as a key molecule in the cross talk between reelin and P15692 signaling pathways . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . 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 . 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 . Q8NG57 a putative tumor suppressor gene of small intestinal neuroendocrine tumors . Small intestinal neuroendocrine tumors ( SI-NETs ) , formerly known as midgut carcinoids , are rare and slow-growing neoplasms . Frequent loss of one copy of chromosome 18 in primary tumors and metastases has been observed . The aim of the study was to investigate a possible role of Q8NG57 ( Elongin A3 ) , currently the only imprinted gene on chromosome 18 , as a tumor suppressor gene in SI-NETs , and whether its expression is epigenetically regulated . Primary tumors , metastases , the human SI-NET cell line CNDT2.5 , and two other cell lines were included . Immunohistochemistry , gene copy number determination by PCR , colony formation assay , western blotting , real-time quantitative RT-PCR , RNA interference , and quantitative CpG methylation analysis by pyrosequencing were performed . A large majority of tumors ( 33/43 ) showed very low to undetectable Elongin A3 expression and as expected 89 % ( 40/45 ) displayed one gene copy of Q8NG57 . The DNA hypomethylating agent DB01262 induced Q8NG57 expression in CNDT2.5 cells , in primary SI-NET cells prepared directly after surgery , but not in two other cell lines . Also siRNA to P26358 and treatment with the general histone methyltransferase inhibitor 3-deazaneplanocin A induced Q8NG57 expression in a cell type-specific way . CpG methylation at the Q8NG57 promoter was observed in all analyzed tissues and thus not related to expression . Overexpression of Q8NG57 resulted in a 50 % decrease in clonogenic survival of CNDT2.5 cells , but not of control cells . The results support a putative role of Q8NG57 as a tumor suppressor gene in SI-NETs . Epigenetic repression of Q8NG57 seems to be tumor cell type-specific and involves both DNA and histone methylation . Alcohols inhibit N-methyl-D-aspartate receptors via a site exposed to the extracellular environment . N-Methyl-D-aspartate ( DB01221 ) receptors are important CNS target sites of alcohols , but the site and mechanism of action of alcohols on DB01221 receptors remains unclear . In CHO- P04264 cells transfected with Q9UHB4 / Q13224 DB01221 receptor subunits , ethanol inhibited DB01221 -activated current with an IC(50) of 138 mM . Truncation of the intracellular C-terminal domain of the Q9UHB4 subunit ( NR1T ) did not alter ethanol sensitivity when combined with the Q13224 subunit , but a similar truncation of the Q13224 subunit ( NR2BT ) slightly enhanced ethanol sensitivity of receptors formed from coexpression with either Q9UHB4 or NR1T subunits . 1-Pentanol applied externally inhibited DB01221 receptors with an IC(50) of 9.9 mM , but intracellular application of 1-pentanol ( 25 mM ) did not alter DB01221 receptor inhibition by externally applied ethanol or 1-pentanol . In addition , the amplitude of DB01221 -activated current did not decrease during the time required for 1-pentanol ( 25 mM ) to diffuse throughout the cytoplasm . DB00898 did not inhibit DB01221 receptors when bath-applied in cell-attached patches or when applied to the cytoplasmic face of inside-out membrane patches . These results appear to be best explained by an action of alcohols on the DB01221 receptor-channel protein , at a site located in a domain exposed to , or only accessible from , the extracellular environment . 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 . Human immunodeficiency virus ( HIV ) -induced neurotoxicity : roles for the DB01221 receptor subtypes . Neuronal damage in human immunodeficiency virus type 1 ( HIV-1 ) infection in the brain is thought to occur at least in part through DB01221 receptor ( NMDAR ) excitation initiated by soluble neurotoxins from HIV-infected brain macrophages . Furthermore , brain regions enriched in NMDAR-2A ( Q12879 ) and NMDAR-2B ( Q13224 ) subunits , such as the hippocampus , are particularly vulnerable . Using cultured rat hippocampal cells and HIV-1-infected human monocyte-derived macrophages ( HIV/MDM ) , we examined the role of Q12879 and Q13224 in HIV/MDM-induced hippocampal neuronal death . We used the primary HIV-1 strain Jago derived from the P04141 of an individual with HIV-associated dementia and that robustly replicates in MDM . We found the following : ( 1 ) hippocampal neuronal susceptibility to HIV/MDM excitotoxins varies according to the developmental expression patterns of Q12879 and Q13224 ; ( 2 ) NMDAR activation by HIV/MDM results in neuronal calpain activation , which results in neuronal death ; and ( 3 ) selective antagonists of homomeric Q13224 / Q13224 - and heteromeric Q12879 / Q13224 -containing NMDARs , as well as an inhibitor of calpain activity , afford neuroprotection against HIV/MDM . These studies establish a clear link between macrophage HIV infection , neuronal Q12879 and Q13224 activation , and calpain-mediated hippocampal neuronal death . They further suggest a dominant role for Q12879 and Q13224 in determining neuronal susceptibility in HIV-infected brain . Antagonists of Q12879 and Q13224 subunits as well as inhibitors of calpain activation offer attractive neuroprotective approaches against HIV in both developing and mature brain . Effect of endothelin inhibition on lung fibroblasts on patients with systemic sclerosis . AIM : The aim of the study was to investigate the effect of selective P25101 DB06268 on viability and differentiation into myofibroblasts of lung fibroblasts derived from SSc-ILD patients and the ability of this drug to modify the lung fibroblast synthesis of P15692 , type I collagen and fibronectin . METHODS : Primary human lung fibroblast cultures were obtained from BAL of SSc-ILD patients . Cell cultures were exposed for 48 h to crescent concentrations of DB06268 ( 10 -6M to 10 -4M ) . In these experimental conditions we evaluated cell viability through crystal violet staining , the production and mRNA expression of P15692 , fibronectin and type I collagen respectively through ELISA and real-Time PCR . Further , we detected alpha-Smooth Muscle Actin ( α-SMA ) through immunocytochemical assay . RESULTS : The lowest concentration of sitaxsentan ( 10-6M ) did not affect fibroblasts viability ; conversely at higher concentrations , sitaxsentan induced a significant inhibition of cell viability . Synthesis and mRNA expression of P15692 , type 1 collagen and fibronectin were significantly reduced in treated lung fibroblasts compared to the untreated ones , in a dose-dependent manner . At higher concentrations , DB06268 reduced the expression of α-SMA . CONCLUSION : The results of this study show that sitaxentan is able in vitro to reduce both cell viability than production of P15692 and extra-cellular matrix components in SSc lung fibroblasts , confirming the anti-fibrotic potential of P25101 in SSc . The decreased expression of α-SMA in treated cells indicate that sitaxsentan may inhibit the fibroblast differentiation toward a myo-fibroblast phenotype and further support the hypothesis that the selective ETRAs may be beneficial in patients with SSc-ILD as anti fibrotic agents . 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 .

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interacts_with DB09068?

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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 . 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 . Analysis of SNP profiles in patients with major depressive disorder . The present study focused on 91 single-nucleotide polymorphisms ( SNPs ) in 21 candidate genes to find associations with major depressive disorder ( MDD ) . In total , 160 healthy controls and 177 patients with MDD were studied . We applied arrayed primer extension ( P27695 ) based genotyping technology followed by association and haplotype analysis . SNPs in P32238 , P21728 , P14416 , and P28335 genes showed nominally significant associations with MDD . None of these associations remained significant after adjustment for multiple testing . Haplotype analysis revealed P32238 haplotypes to be associated with MDD ( global p=0.004 ) . More precisely , we found the GAGT haplotype to be associated with increased risk for MDD ( OR 7.42 , 95 % CI 2.13-25.85 , p=0.002 ) . This haplotype effect remained significant after Bonferroni correction ( p=0.04 after Bonferroni 's adjustment ) . Altogether we were able to find some nominal associations , but due to small sample size these results should be taken as exploratory . However , the effect of GAGT haplotype on the P32238 gene may be considered as increasing the risk for MDD . Recombinant human prothrombin kringles have potent anti-angiogenic activities and inhibit Lewis lung carcinoma tumor growth and metastases . P00734 , a protein involved in blood coagulation , is a plasma glycoprotein composed of the Gla domain , two adjacent kringle domains , and a serine protease domain . Kringles are triple-disulfide-loop folding domains , which are found in several other blood proteins . In this study , we showed that recombinant human prothrombin kringle-1 , -2. and -1-2 ( rk-1 , -2 , -1-2 ) all have potent anti-angiogenic activities , which inhibit Lewis lung carcinoma ( LLC ) tumor growth and metastases . Recombinant human prothrombin kringles were expressed by an E. coli expression system and purified to apparent homogeneity from crude E. coli extracts . Purified rk-1 , -2 , -1-2 migrated with a molecular mass of 14 , 19 , and 31 kDa , respectively , on sodium dodecyl sulfate-polyacrylamide gel electrophoresis ( SDS-PAGE ) under reducing conditions . rk-1 , -2 , -1-2 exhibited potent inhibitory effects on P09038 -stimulated bovine capillary endothelial cell growth with half-maximal concentrations ( ED50 ) of approximately 41 , 55 , and 156 nM , respectively . All of the recombinant human prothrombin kringles also inhibited angiogenesis in the chorioallantoic membrane ( P62158 ) of chick embryos at a dose of 20 microg . Systemic administration of rk-1 , -2 , -1-2 at a dose of 0.5 mg/kg/day suppressed the growth of primary LLC and at dose of 0.5 and 1.0 mg/kg/day inhibited LLC metastases in C57BL6/J mice lungs through their anti-angiogenic effects . Autophagy suppression promotes apoptotic cell death in response to inhibition of the PI3K- P42345 pathway in pancreatic adenocarcinoma . Targeting of pathways downstream of DB01367 represents a promising therapeutic strategy for pancreatic cancer , the fourth leading cause of cancer-related death in the USA , since activation of the Raf-MEK- P29323 and PI3K-AKT pathways is found frequently in this disease and is associated with poor prognosis . Taking advantage of a panel of human PDAC cell lines and specific inhibitors of PI3K and/or P42345 , we systematically address the question whether dual-targeted inhibition of the PI3K and P42345 pathways offers advantages over single-targeted inhibition of PI3K in PDAC . We observe greater overall susceptibility of cell lines to dual inhibition compared to targeting PI3K alone . However , we find that dual inhibition of PI3K and P42345 induces autophagy to a greater extent than inhibition of each target alone . In agreement with this , we show that combined administration of PI3K/ P42345 and autophagy inhibitors results in increased anti-tumor activity in vitro and in vivo in models of pancreatic adenocarcinoma . DB05241 , a PI3K/ P42345 inhibitor used in our in vivo studies , is currently undergoing clinical evaluation in a variety of cancer types , while the autophagy inhibitor chloroquine is a widely used anti-malaria compound . Thus , our studies provide rationale for clinical development of combinations of these compounds for the treatment of pancreatic adenocarcinoma . 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 . 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 . 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 . P15121 regulates high glucose-induced ectodomain shedding of tumor necrosis factor ( P01375 ) -alpha via protein kinase C-delta and P01375 converting enzyme in vascular smooth muscle cells . Chronic low-grade inflammation has emerged as a key contributor to the cardiovascular complications of diabetes , however , the mechanisms by which diabetes increases inflammation remain poorly understood . Here , we report that exposure to high glucose ( HG ) stimulates ectodomain shedding of P01375 from rat aortic smooth muscle cells in culture . Our results show that exposure to HG decreases membrane-associated P01375 . This decrease in unprocessed P01375 was prevented by the aldose reductase ( AR ) inhibitor sorbinil and AR small interference RNA . Treatment with HG , but not equimolar mannitol or 3-O-methyl glucose , resulted in phosphorylation and activation of P01375 converting enzyme ( P78536 ) ( P78536 ) , which were attenuated by sorbinil or AR-specific small interference RNA . HG-induced P78536 phosphorylation and P01375 processing were also prevented by P01375 protease inhibitor-1 , an inhibitor of P78536 . Inhibition of protein kinase C ( PKC ) -delta by rottlerin prevented HG-induced P78536 activation and the accumulation of unprocessed P01375 . Treatment with sorbinil decreased elevated levels of circulating P01375 in streptozotocin-treated diabetic rats . DB02712 treatment also decreased the expression of P01375 , matrix metalloproteinase-2 , matrix metalloproteinase-9 , and increased tissue inhibitor of metalloproteinase-3 in vascular smooth muscle cells treated with HG and in balloon-injured carotid arteries of diabetic rats . These results indicate that HG-induced P01375 shedding could be attributed to P78536 activation , which is regulated , in part , by PKC-delta and AR . Therefore , inhibition of P78536 by P01375 protease inhibitor-1 , or pharmacological inhibition of PKC-delta or AR may represent useful strategies for treating vascular inflammation associated with diabetes . 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 . 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 . Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 , CD8 and forkhead box P09131 ( Foxp3 ) , respectively . P14136 , Iba1 and P34810 -immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed . Inhibition of phosphatidylserine biosynthesis in developing rat brain by maternal exposure to ethanol . DB00144 ( PtdSer ) , major acidic phospholipids in neuronal membranes , participate in important cell signaling processes . The PtdSer in brain is highly enriched with docosahexaenoic acid ( DB01708 ; 22:6n-3 ) , and the DB01708 status or ethanol exposure has been shown to influence the PtdSer level . This study shows that ethanol exposure during prenatal and developmental period significantly attenuates microsomal PtdSer biosynthetic activities and reduces PtdSer , particularly 18:0 , 22:6-PtdSer , in developing rat brain cortices . Brain microsomes were incubated with deuterium labeled exogenous substrates in vitro and the products formed were detected by reversed phase HPLC-electrospray ionization mass spectrometry ( P19957 -MS ) . These in vitro bioassays showed that 1-stearoyl-2-docosahexaenoyl ( 18:0 , 22:6 ) species is the best substrate for PtdSer synthesis from both phosphatidylcholine ( PtdCho ) and phosphatidylethanolamine ( PtdEtn ) . The PtdSer biosynthetic activity of brain , especially for 18:0 , 22:6-PtdSer production , was hampered significantly by maternal exposure to ethanol . PtdSer levels were consistently reduced significantly in brain cortices of the pups from ethanol-exposed dams , due mainly to the depletion of 18:0 , 22:6-PtdSer . The mRNA expression of P48651 ( PSS1 ) and Q9BVG9 ( Q9BVG9 ) was not reduced by ethanol . Similarly , the PSS1 enzyme level did not change after ethanol exposure but Q9BVG9 could not be probed with the antibody available currently . Degradation of PtdSer by mitochondrial PtdSer decarboxylation was not enhanced but also inhibited . Taken together , attenuated PtdSer biosynthetic activities are largely responsible for the PtdSer reduction observed in developing rat brains after maternal exposure to ethanol . Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . P08908 receptor-mediated regulation of mitogen-activated protein kinase phosphorylation in rat brain . Mitogen-activated protein kinases ( MAPKs ) , a family of signal transduction mediators important in a host of cellular activities , include the extracellular signal-regulated kinases Erk1 and Erk2 . We determined whether 5-HT(1A) receptors activate Erk1/2 in rat brain in vivo , as they do in recombinant cell lines . In contrast to the effect in cells , the 5-HT(1A) receptor agonist 8-hydroxy-N,N-diproylaminotetralin ( 8-OH-DPAT ) dose- and time-dependently decreased basal levels of phosphorylated Erk1/2 ( phospho-Erk1/2 ) in rat hippocampus ( ED(50) approximately 0.1 mg/kg , maximum approximately 90 % ) without altering total Erk1/2 . The effects were kinase-specific , as 8-OH-DPAT did not modify phosphorylated or total levels of the MAPKs c-Jun-N-terminal kinase/stress-activated protein kinase ( JNK/SAPK ) and p38 MAPK . Moreover , 8-OH-DPAT did not modify phospho-Erk1/2 in striatum or frontal cortex . The effect of 8-OH-DPAT was blocked by pretreatment with the selective 5-HT(1A) receptor antagonists N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinylcyclohexanecarboxamide ( WAY 100635 ) , 1-(2-methoxyphenyl)-4-(4-[2-phthalimido]butyl)piperazine ( NAN-190 ) and 4-fluoro-N-(2-[4-(2-methoxyphenyl)1-piperazinyl]ethyl)-N-(2-pyridinyl)benzamide dihydrochloride ( p-MPPF ) , but not by the weak partial agonist/antagonist 8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro(4.5)decane-7,9-dione dihydrochloride ( BMY 7378 ) . Other 5-HT(1A) receptor agonists ( buspirone , gepirone and ipsapirone ) also reduced phospho-Erk1/2 levels in hippocampus . 8-OH-DPAT also reduced the levels of the upstream activator of Erk1/2 , phosphorylated extracellular signal-regulated kinase kinase ( phospho- Q02750 /2 ) , and at least one potential downstream target , the nuclear transcription factor phospho-Elk-1 . The region- and kinase-specific effects suggest that the Erk1/2 signal transduction cascade is likely an important differential mediator of 5-HT(1A) receptor-regulated events in the central nervous system . Dabrafenib and trametinib , alone and in combination for P15056 -mutant metastatic melanoma . Dabrafenib and trametinib were approved for use as monotherapies in P15056 -mutant metastatic melanoma by the U.S . Food and Drug Administration ( FDA ) in 2013 , and most recently , their use in combination has received accelerated FDA approval . Both drugs target the mitogen-activated protein kinase ( MAPK ) pathway : dabrafenib selectively inhibits mutant P15056 that constitutively activates the pathway , and trametinib selectively inhibits Q02750 and P36507 proteins activated by RAF kinases . The phase III study of dabrafenib in P15056 (V600E) metastatic melanoma reported rapid tumor regression in most patients and a 59 % objective RECIST response rate . The median progression-free survival ( PFS ) and overall survival ( OS ) were improved compared with dacarbazine . Toxicities were well tolerated and different from those reported for vemurafenib , the first FDA-approved P15056 inhibitor . Efficacy has been demonstrated in other P15056 -mutant genotypes . The phase III study of trametinib in P15056 inhibitor-naïve patients with P15056 (V600E) or P15056 (V600K) also showed benefit with a prolonged median PFS and OS compared with chemotherapy . DB08911 is ineffective in patients who have progressed on P15056 inhibitors . A phase II trial of combined dabrafenib and trametinib demonstrated higher response rates and longer median PFS than dabrafenib monotherapy , with less cutaneous toxicity . Here , we review the clinical development of both drugs as monotherapies and in combination , and discuss their role in the management of P15056 -mutant melanoma . P02751 and VLA-4 in haematopoietic stem cell-microenvironment interactions . The self-renewal and differentiation of haematopoietic stem cells occurs in vivo and in vitro in direct contact with cells making up the haematopoietic microenvironment . In this study we used adhesive ligands and blocking antibodies to identify stromal cell-derived extracellular matrix proteins involved in promoting attachment of murine haematopoietic stem cells . Here we report that day-12 colony-forming-unit spleen (CFU- P28222 )5 cells and reconstituting haematopoietic stem cells attach to the C-terminal , heparin-binding fragment of fibronectin by recognizing the P28290 peptide of the alternatively spliced non-type III connecting segment ( IIICS ) of human plasma fibronectin . Furthermore , CFU- P28222 stem cells express the alpha 4 subunit of the VLA-4 integrin receptor , which is known to be a receptor for the P28290 sequence , and monoclonal antibodies against the integrin alpha 4 subunit of VLA-4 block adhesion of CFU- P28222 stem cells to plates coated with the C-terminal fibronectin fragment . Finally , polyclonal antibodies against the integrin beta 1 subunit of VLA-4 inhibit the formation of CFU- P28222 -derived spleen colonies and medullary haematopoiesis in vivo following intravenous infusion of antibody-treated bone marrow cells . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . Use of the ImmuKnow assay to evaluate the effect of alemtuzumab-depleting induction therapy on cell-mediated immune function after renal transplantation . BACKGROUND : Good outcomes after renal transplantation are dependent on effective immunosuppression while minimizing infection . DB00087 ( Campath or Campath-1H ) is an anti- P31358 humanized monoclonal IgG1 antibody which induces rapid and sustained depletion of circulating lymphocytes and has been effectively used as an immunosuppressant in post-transplant induction therapy . METHODS : We used the ImmuKnow assay to compare cell-mediated immune function in renal transplant patients treated with alemtuzumab or with conventional immunosuppressive tri-therapy . The ImmuKnow method determines the levels of adenosine triphosphate ( DB00171 ) released from P01730 cells following stimulation with a mitogen . RESULTS : We showed a statistically significant difference in the distribution of outcome after transplantation between the conventional and the Campath groups ( P = 0.010 ) . A significantly higher number of patients treated with alemtuzumab induction therapy were stable after transplantation compared to those treated with conventional immunosuppressive tri-therapy ( 96.6 vs. 75.7 % ) . DB00171 values were significantly higher in the conventional group compared to the Campath group at 180 days after transplantation ( P < 0.001 ) . DB00171 levels did not change significantly over time in clinically stable kidney recipients treated with alemtuzumab induction therapy ( P = 0.554 ) . CONCLUSIONS : The ImmuKnow assay is a useful tool for evaluating the global immune response in alemtuzumab-treated renal transplant patients . DB00087 -depleting induction therapy remains effective for at least 180 days . Inhibition of noradrenaline release via presynaptic P28222 receptors of the rat vena cava . In the rat inferior vena cava preincubated with 3H-noradrenaline , the effects of nine serotonin ( 5-HT ) receptor agonists and of eight antagonists ( including two beta-adrenoceptor blocking agents ) on the electrically evoked 3H overflow were determined . 1 . 5-HT , 5-carboxamido-tryptamine , 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole ( RU 24969 ) , 5-methoxytryptamine , N,N-dimethyl-5HT , tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow . The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for P28222 binding sites , but not with their affinities for P08908 , P28335 or 5-HT2 binding sites . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 receptor agonist , and ipsapirone , a partial agonist at these receptors , did not inhibit overflow . 2 . Cyanopindolol facilitated the evoked 3H overflow , an effect which was abolished by propranolol . The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol . 3 . The concentration-response curve for 5-HT was shifted to the right by metitepine , metergoline , quipazine , 6-chloro-2-(1-piperazinyl)pyrazine ( MK 212 ) and propranolol which , given alone , did not affect 3H overflow . The apparent pA2 values of these antagonists tended to be correlated with their affinities for P28222 ( but not P08908 , P28335 or 5-HT2 ) binding sites . Ketanserin , a 5-HT2 receptor antagonist , and spiperone , which blocks 5-HT2 and P08908 but not P28222 or P28335 receptors , failed to antagonize the effect of 5-HT. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 .

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Pharmacokinetic profiles of the novel P35354 selective inhibitor cimicoxib in dogs . DB05095 ( CX ) is a novel imidazole derivative that is a cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drug and the latest P35354 selective inhibitor to be released for veterinary use . Currently there is limited information available on the pharmacokinetic ( PK ) properties of CX . The aim of the current study was to evaluate the PK features of CX after administration of the recommended dose and after administration of a more variable dose rate in the form of the commercially available tablet . In addition , the effects of food intake on the PK properties were also evaluated . In the first study , five healthy Beagle dogs received 2mg/kg CX via the oral route following a period of fasting . The second study was conducted using six healthy Labrador retriever dogs which each received an 80 mg tablet ( approximate dose 1.95-2.5mg/kg ) using a crossover design , both in the fasted and fed condition . The plasma concentrations of CX were detected by a validated HPLC method . No adverse effects were observed in any dogs during the experiment . The results from the PK analysis were similar between the studies , regardless of precision of dose and fasted and fed conditions . The mean peak concentration of CX was 0.49 and 0.43 μg/mL under fasted and fed conditions , respectively . The mean half-life was about 3h after all treatments . In addition , simulated multiple dosing data revealed that time over minimal effective concentration was similar after 1.95 , 2.0 and 2.5mg/kg dose administrations . These findings suggest that slight variation from the recommended dose should not alter the therapeutic outcome . In addition , CX can be administered to fed dogs without significantly affecting blood levels . Inhibition of the T790M gatekeeper mutant of the epidermal growth factor receptor by EXEL-7647 . PURPOSE : Agents inhibiting the epidermal growth factor receptor ( P00533 ) have shown clinical benefit in a subset of non-small cell lung cancer patients expressing amplified or mutationally activated P00533 . However , responsive patients can relapse as a result of selection for P00533 gene mutations that confer resistance to DB00171 competitive P00533 inhibitors , such as erlotinib and gefitinib . We describe here the activity of EXEL-7647 ( DB05007 ) , a novel spectrum-selective kinase inhibitor with potent activity against the P01133 and vascular endothelial growth factor receptor tyrosine kinase families , against both wild-type ( WT ) and mutant P00533 in vitro and in vivo . EXPERIMENTAL DESIGN : The activity of P00533 inhibitors against WT and mutant EGFRs and their effect on downstream signal transduction was examined in cellular assays and in vivo using A431 and MDA-MB-231 ( WT P00533 ) and H1975 ( L858R and T790M mutant P00533 ) xenograft tumors . RESULTS : EXEL-7647 shows potent and long-lived inhibition of the WT P00533 in vivo . In addition , EXEL-7647 inhibits cellular proliferation and P00533 pathway activation in the erlotinib-resistant H1975 cell line that harbors a double mutation ( L858R and T790M ) in the P00533 gene . In vivo efficacy studies show that EXEL-7647 substantially inhibited the growth of H1975 xenograft tumors and reduced both tumor P00533 signaling and tumor vessel density . Additionally , EXEL-7647 , in contrast to erlotinib , substantially inhibited the growth and vascularization of MDA-MB-231 xenografts , a model which is more reliant on signaling through vascular endothelial growth factor receptors . CONCLUSIONS : These studies provide a preclinical basis for clinical trials of DB05007 in solid tumors and in patients bearing tumors that are resistant to existing P00533 -targeted therapies . DB04864 , but not tacrine , stimulates P04271 secretion in astrocyte cultures . AIMS : The loss of cholinergic function in the central nervous system contributes significantly to the cognitive decline associated with advanced age and dementias . DB04864 ( HupA ) is a selective inhibitor of acetylcholinesterase ( P22303 ) and has been shown to significantly reduce cognitive impairment in animal models of dementia . Based on the importance of astrocytes in physiological and pathological brain activities , we investigated the effect of HupA and tacrine on P04271 secretion in primary astrocyte cultures . P04271 is an astrocyte-derived protein that has been proposed to be a marker of brain injury . MAIN METHODS : Primary astrocyte cultures were exposed to HupA , tacrine , cholinergic agonists , and P04271 secretion was measured by enzyme-linked immunosorbent assay ( ELISA ) at 1 and 24h . KEY FINDINGS : HupA , but not tacrine , at 100μM significantly increased P04271 secretion in astrocyte cultures . DB00184 ( at 100 and 1000μM ) was able to stimulate P04271 secretion in astrocyte cultures . SIGNIFICANCE : Our data reinforce the idea that P22303 inhibitors , particularly HupA , do not act exclusively on the acetylcholine balance . This effect of HupA could contribute to improve the cognitive deficit observed in patients , which are attributed to cholinergic dysfunction . In addition , for the first time , to our knowledge , these data indicate that P04271 secretion can be modulated by nicotinic receptors , in addition to glutamate , dopamine and serotonin receptors . 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 . Inhibition of the invasion capacity of carcinoma cells by DB05476 , a novel synthetic inhibitor of the urokinase-type plasminogen activator system . The overall survival rate of patients suffering from carcinomas has remained poor and nearly unchanged over the last decades . This is mainly due to the so-called minimal residual disease , i.e. , remaining tumor cells that overcome surgery and/or radiotherapy and are the cause of locoregional and distant metastases . To metastasize , tumor cells take advantage of proteases to invade and remodel surrounding tissues . Here , we analyzed the efficiency of DB05476 , a novel 3-amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 expression was characterized in different carcinoma cell lines , including SCCHN , breast and cervical carcinoma . Thereafter , the invasive potential of these cell lines was determined using Matrigel invasion chambers and a spheroid cocultivation model with human fibroblasts . Q03405 expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 . A decrease of tumor cell invasion by up to 50 % was achieved in both models with the SCCHN line FaDu and the cervical carcinoma line HeLa after treatment with DB05476 . Thus , our results demonstrate the potential of DB05476 in vitro as a promising adjuvant antimetastatic therapy of carcinomas . 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 . 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 . Phytoestrogens induce apoptosis through a mitochondria/caspase pathway in human breast cancer cells . OBJECTIVE : To explore the effect and pathway of phytoestrogens on the growth of breast cancer cell line MCF-7 . METHODS : MCF-7 cells ( human estrogen receptor-positive and progesterone receptor-positive breast cancer cells ) were cultured in serum-free medium for 24 h and then treated with genistein , resveratrol , and quercetin ( 10(-10)-10(-4) mol/l ) . After further incubation for 24 , 48 , 72 , and 92 h , the cells were harvested and extracted for 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ( MTT ) assay . According to the above results , the proteins involving proliferative and apoptotic pathways were evaluated by Western blot analysis . RESULTS : DB01645 , resveratrol , and quercetin significantly inhibited cellular proliferation in a dose- and time-dependent manner . Significantly elevated caspase-3 activity was noted after treatment with genistein ( 10(-9)-10(-7) mol/l ) , as well as with resveratrol and quercetin ( 10(-9)-10(-5) mol/l ) . Significant reduction of PI3K and AKT protein and significant increase of P48023 , Fas-associated protein with death domain , cytochrome C , truncated Bid , caspase-9 , and caspase-3 were all noted after genistein , resveratrol , and quercetin treatment . 17β-Estradiol induced completely opposite results . P03372 ( ER ) α expression was significantly increased with 17β-estradiol , whereas ERβ expression was significantly elevated in the cultures with genistein , resveratrol , and quercetin . CONCLUSIONS : These data demonstrate that genistein , resveratrol , and quercetin have antiproliferative effects on breast cancer cells . Their induction of apoptosis involves the activation of both the intrinsic and extrinsic apoptotic pathways , which may be related to the differential affinity to ERα and ERβ . Whether phytoestrogens have similar effects on normal breast cells remains to be examined . Encapsulation in lipid-core nanocapsules overcomes lung cancer cell resistance to tretinoin . Tretinoin is a retinoid derivative that has an antiproliferative effect on several kinds of tumours . Human lung adenocarcinoma epithelial cell lines ( A549 ) exhibit a profound resistance to the effects of tretinoin . Nanocarriers seem to be a good alternative to overcomecellular resistance to drugs . The aim of this study was to test whether tretinoin-loaded lipid-core nanocapsules exert anantitumor effect on A549 cells . A549 cells were incubated with free tretinoin ( Q8WZ42 ) , blank nanocapsules ( LNC ) and tretinoin-loaded lipid-core nanocapsules ( Q8WZ42 -LNC ) . Data from evaluation of DNA content and P08758 binding assay by flow cytometry showed that Q8WZ42 -LNC induced apoptosis and cell cycle arrest at the P55008 -phase while Q8WZ42 did not . Q8WZ42 -LNC showed higher cytotoxic effects than Q8WZ42 on A549 cells evaluated by MTT and LIVE/DEAD cell viability assay . Gene expression profiling identified up-regulated expression of gene P38936 by Q8WZ42 -LNC , supporting the cell cycle arrest effect . These results showed for the first time that Q8WZ42 -LNC are able to overcome the resistance of adenocarcinoma cell line A549 to treatment with Q8WZ42 by inducing apoptosis and cell cycle arrest , providing support for their use in applications in lung cancer therapy . P10275 as a therapeutic target . Androgens function as sex hormone primarily via activation of a single androgen receptor ( AR , or P10275 ) . AR is an important therapeutic target for the treatment of diseases such as hypogonadism and prostate cancer . AR ligands of different chemical structures and/or pharmacological properties are widely used for these therapeutic applications , and all of the AR ligands currently available for therapy modulate AR function via direct binding to the ligand-binding pocket ( P18428 ) of the receptor . In the past ten years , our understanding of AR structure and molecular mechanism of action has progressed extensively , which has encouraged the rapid development of newer generation of AR ligands , particularly tissue-selective AR ligands . With improved tissue selectivity , future generations of AR ligands are expected to greatly expand the therapeutic applications of this class of drugs . This review will provide an overview of the common therapeutic applications of currently available AR ligands , and discussion of the major challenges as well as novel therapeutic strategies proposed for future drug development . Detection of multiple antibodies in myasthenia gravis and its clinical significance . BACKGROUND : Antibodies against acetylcholine receptor , acetylcholinesterase , ryanodine receptor and titin have been found in patients with myasthenia gravis . However , the relations between these antibodies and character of myasthenia gravis are unknown . This study aimed to detect multiple antibodies in myasthenia gravis and to investigate its clinical significance . METHODS : These antibodies were detected by enzyme-linked immunoabsorbent assay in 89 cases of myasthenia gravis , 66 cases of other neurological diseases and 66 healthy controls . The incidences of antibodies were compared using the chi-square test . RESULTS : DB03128 receptor , acetylcholinesterase , titin and ryanodine receptor antibodies were detected in 53.9 % , 20.2 % , 64.0 % and 55.0 % of myasthenia gravis patients respectively , higher than in patients of other neurological diseases and controls groups . The combination of the four antibodies assays provided 94.4 % sensitivity and 84.0 % specificity for the diagnosis of myasthenia gravis . P22303 antibody occurred more frequently in acetylcholine receptor antibody negative patients with adverse reactions to neostigmine test . Q8WZ42 antibody provided 82.1 % sensitivity and 52.5 % specificity for myasthenia gravis with thymoma . Incidences of titin and of ryanodine receptor antibody were higher in late onset myasthenia gravis than in early onset myasthenia gravis . The proportion of titin antibody positive patients increased with the severity of myasthenia gravis as graded by a modified Osserman scale . CONCLUSIONS : Testing for acetylcholine receptor , acetylcholinesterase , titin and ryanodine receptor antibodies can offer a better diagnostic method for myasthenia gravis than each antibody test alone . Q8WZ42 antibody combined with computed tomography was better for the diagnosis of thymoma . Q8WZ42 antibody occurred most frequently in severe myasthenia gravis . 5-Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5-Azacitine ( 5-Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5-Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 , phospho- Q13158 , p16(INKA) , Bax , Bak , and P38936 ( P38936 ) , and inhibited FLIP , Bcl-2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5-Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5-Aza with AR-antagonist DB01128 had additive/synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5-Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC . Inhibition of p53 transcriptional activity by the P04271 calcium-binding protein . The levels of S100 Ca(2+)-binding proteins correlate with the progression of certain tumors , but their role , if any , in carcinogenesis is still poorly understood . P04271 protein associates with both the p53 oligomerization domain ( residues 325-355 ) and the extreme C terminus of the tumor suppressor p53 ( residues 367-392 ) . Consequently , P04271 inhibits p53 tetramer formation and p53 phosphorylation mediated by protein kinase C , on p53 C-terminal end . In this report , we show that the P04271 protein decreases p53 DNA binding and transcriptional activity . The effect of P04271 is reflected in vivo by a reduced accumulation of p53 , P38936 , and Q00987 protein levels in co-transfection assays and in response to bleomycin . The P04271 can still interact with p53 in the absence of p53 extreme C-terminal end and reduce the expression of p53 downstream effector genes . These data indicate that P04271 does not require p53 extreme C-terminal end to inhibit p53 activity . Collectively , these findings imply that elevated levels of P04271 in tumors such as astrocytomas and gliomas could inhibit p53 functions and contribute to cancer progression . 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 . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . 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 . 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 . Human rheumatoid synoviocytes express functional Q99572 receptors . Human type B synoviocytes are involved in joint injury during rheumatic diseases by producing inflammatory mediators such as interleukin-6 ( P05231 ) . The increased level of purine and pirimidine nucleotides in the synovial fluid of rheumatoid arthritis ( RA ) patients could activate the large family of P2 receptors . Thus , we investigated the presence of P2 receptors in human type B synoviocytes from rheumatoid joints , also evaluating whether the Q99572 receptor is involved in P05231 release . Reverse transcriptase polymerase chain reaction analysis revealed messenger ribonucleic acid ( mRNA ) expression for the P51575 , Q9UBL9 , Q99571 , Q93086 , O15547 , Q99572 , P47900 , P51582 , Q96G91 , Q9H244 , Q9BPV8 , and Q15391 but not the P56373 , P41231 , and Q15077 receptors . The expression of the Q99572 receptor was confirmed by Western blot analysis . DB00171 ( DB00171 ) and the Q99572 receptor agonist 2'-3'-O-(4-benzoylbenzoyl) DB00171 ( BzATP ) triggered an increase in intracellular calcium , thereby suggesting the expression of functional P2 receptors , including the Q99572 receptor . Moreover , BzATP treatment upregulated both P05231 mRNA and protein expression . Synoviocytes spontaneously released low quantities of P05231 ; the incubation with BzATP induced the release of larger amounts of the cytokine , and such a release was blunted by the Q99572 antagonist oxidized DB00171 . The selective P51575 and P56373 receptor agonist alpha,beta-methylene DB00171 did not affect P05231 release . Finally , BzATP failed to induce a significant uptake of the large-molecule YO-PRO , thus suggesting the lack of pore formation after Q99572 receptor stimulation . In conclusion , among the different P2 receptors expressed on human RA type B synoviocytes , the Q99572 receptor may modulate P05231 release but not inducing changes in cell membrane permeability . 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 . Modulation of leukocyte transendothelial migration by integrin-associated glycosyl phosphatidyl inositol ( P06744 ) -anchored proteins . Leukocyte transendothelial migration is an essential process in inflammation and the immune response . The mechanisms involved in leukocyte adhesion to the endothelium , forming the first step in leukocyte extravasation , have been fairly well documented . However , subsequent steps , which include de-adhesion , coupled with locomotion , remain largely unknown . As part of our efforts to study leukocyte transendothelial migration , we previously established a monoclonal antibody ( mAb ) that sequentially up-regulates and down-regulates beta2 integrin-dependent adhesion of human neutrophils , as well as transendothelial migration in vitro . The molecule recognized by this mAb is a glycosyl phosphatidyl inositol , ( P06744 ) -anchored glycoprotein . This protein may prove to be a new member of the family of integrin-associated , P06744 -anchored proteins , which includes urokinase-type plasminogen activator receptor ( Q03405 ) , lipopolysaccharide ( LPS ) /LPS binding protein ( P18428 ) receptor ( P08571 ) , and Fcgamma receptor IIIB ( CD16b ) ; all of which are regulators of integrin function . The mechanisms involved in beta2 integrin regulation by this new P06744 -anchored glycoprotein are discussed . Inhibition of Q16552 as a pharmacological approach for Q9UKU7 . Several experimental approaches have been utilized , in order to critically examine the roles of Q16552 family members in intestinal inflammation . These approaches have included : ( 1 ) the use of Q16552 and Q96PD4 -deficient mice , ( 2 ) specific antibodies directed against Q16552 , ( 3 ) an Q16552 vaccine , ( 4 ) methods to block the Q16552 receptor and ( 5 ) small-molecule inhibitors of Q16552 . Previous studies found somewhat conflicting results in preclinical models of Inflammatory Bowel Disease ( Q9UKU7 ) , using specific strains of Q16552 -deficient mice . This paper will review the preclinical results using various pharmacological approaches [ specific Q16552 antibodies , an Q16552 receptor fusion protein , IL-12/IL-23 p40 subunit and Q16552 vaccine approaches , as well as a small molecule inhibitor ( Vidofludimus ) ] to inhibit Q16552 in animal models of Q9UKU7 . Recent clinical results in patients with Q9UKU7 will also be discussed for DB09029 ( an Q16552 antibody ) , Brodalumab ( an Q16552 receptor antibody ) and two small-molecule drugs ( Vidofludimus and DB08895 ) , which inhibit Q16552 as part of their overall pharmacological profiles . This review paper will also discuss some pharmacological lessons learned from the preclinical and clinical studies with anti- Q16552 drugs , as related to drug pharmacodynamics , Q16552 receptor subtypes and other pertinent factors . Finally , future pharmacological approaches of interest will be discussed , such as : ( 1 ) Retinoic acid receptor-related orphan nuclear receptor gamma t ( Rorγt ) antagonists , ( 2 ) P10276 ( RARα ) antagonists , ( 3 ) Pim-1 kinase inhibitors and ( 4 ) Dual small-molecule inhibitors of NF-κB and P40763 , like synthetic triterpenoids .

MH_train_1556

MH_train_1556

MH_train_1556

interacts_with DB00501?

multiple_choice

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 . Natriuretic and renoprotective effect of chronic oral neutral endopeptidase inhibition in acute renal failure . P08473 ( NEP : EC 3.4.24.11 ) is involved in the degradation of peptides such as atrial natriuretic peptide , angiotensin II ( AngII ) , and endothelin-1 ( ET-1 ) . In this study we propose that NEP inhibition provides protection in glycerol-induced acute renal failure ( Q8N726 ) . Renal vascular responses were evaluated in Q8N726 rats where Q8N726 was induced by injecting 50 % glycerol in candoxatril , a NEP inhibitor ( 30 mg/kg , orally ; for 3 weeks ) pretreated rats . AngII and U46619 ( a TxA2 mimetic ) vasoconstriction was increased ( 2- to 4-fold ) in Q8N726 while ET-1 vasoconstriction was surprisingly reduced ( 23+/-3 % ; p < 0.05 ) . In Q8N726 , candoxatril paradoxically enhanced ET-1 response ( 60+/-20 % ; p < 0.05 ) but reduced AngII vasoconstriction ( 51+/-11 % ; p < 0.05 ) without affecting U46619 response . However , candoxatril treatment was without effect on plasma ET-1 and TxB2 levels in Q8N726 . DB00616 reduced plasma AngII by 34+/-4 % ( p < 0.05 ) in Q8N726 which was approximately 3.5-fold higher compared to control . DB00616 doubled the nitrite excretion in control but was without effect on proteinuria or nitrite excretion in Q8N726 . DB00616 enhanced Na+ and creatinine excretion in Q8N726 by 73+/-9 % and 33+/-2 % , respectively . These results suggest that NEP inhibition may confer protection in glycerol-induced Q8N726 by stimulating renal function but without a consistent effect on renal production and renal vascular responses to endogenous vasoconstrictors . 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 . Differential cytokine expression in Chlamydophila psittaci genotype A- , B- or D-infected chicken macrophages after exposure to Escherichia coli O2: P04264 LPS . Chlamydophila ( Cp. ) psittaci and avian pathogenic Escherichia ( E. ) coli infections contribute to the respiratory disease complex observed in turkeys . Secondary infection with E. coli exacerbates Cp. psittaci pathogenicity and augments E. coli excretion . The innate immune response initiated by both pathogens in their avian host is unknown . We therefore determined the cytokine responses following Cp. psittaci infection and E. coli superinfection of avian monocytes/macrophages by examining gene transcripts of IL-1beta , P05231 , CXCLi2 ( P10145 ) , CXCLi1 ( K60 ) , P22301 , IL-12alpha/beta , Q14116 , TGF-beta4 and CCLi2 at 4h post-inoculation with different Cp. psittaci strains or 4h post-treatment with avian E. coli LPS of Cp. psittaci pre-infected Q96DB2 cells . Cp. psittaci strains used were 84/55 and 92/1293 ( highly virulent ) , CP3 ( low virulent ) and 84/2334 ( phylogenetically intermediate between Cp. psittaci and Chlamydophila abortus ) . At 4h post chlamydial infection , an increased expression of IL-1beta and P05231 as well as CXCLi2 , CXCLi1 and CCLi2 was observed compared to levels in uninfected Q96DB2 controls . This effect was less pronounced for the milder CP3 strain . The pro-inflammatory response of Cp. psittaci infected cells to E. coli LPS was significantly lowered compared to uninfected controls , especially when the cells were pre-infected with highly virulent Cp. psittaci strains . In both experiments , exceptionally high P22301 and no TGF-beta4 responses were observed , and we propose that this could induce macrophage deactivation and NF-kappaB suppression . Consequently , pro-inflammatory and Th1-promoting responses to both the primary Cp. psittaci infection and E. coli would be inhibited , thus explaining the observed aggravated in vivo pathology . 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 . Regulation of the natural killer cell response to interferon-alpha by biogenic amines . Monocytes , recovered from human peripheral blood by counter-current centrifugal elutriation ( CCE ) , suppressed baseline natural killer ( NK ) cell cytotoxicity ( NKCC ) and rendered NK cells resistant to activation of cytotoxicity by human recombinant interferon-alpha ( IFN-alpha ) by a cell contact-dependent mechanism . Monocyte-induced suppression of resting and IFN-activated NK cells was abrogated by the biogenic amines histamine [ via H2-type receptors ( P25021 ) ] and serotonin [ via P08908 -type receptors ( 5-HT1AR ) ] . Our data are suggestive of a monocyte/NK cell interaction that is subject to regulation by biogenic amines . Q00535 activator protein p25 preferentially binds and activates GSK3β . Glycogen synthase kinase 3β ( GSK3β ) and cyclin-dependent kinase 5 ( Q00535 ) are tau kinases and have been proposed to contribute to the pathogenesis of Alzheimer 's disease . The 3D structures of these kinases are remarkably similar , which led us to hypothesize that both might be capable of binding cyclin proteins -- the activating cofactors of all CDKs . Q00535 is normally activated by the cyclin-like proteins p35 and p39 . By contrast , we show that GSK3β does not bind to p35 but unexpectedly binds to p25 , the calpain cleavage product of p35 . Indeed , overexpressed GSK3β outcompetes Q00535 for p25 , whereas Q00535 is the preferred p35 partner . FRET analysis reveals nanometer apposition of GSK3β:p25 in cell soma as well as in synaptic regions . Interaction with p25 also alters GSK3β substrate specificity . The GSK3β:p25 interaction leads to enhanced phosphorylation of tau , but decreased phosphorylation of β-catenin . A partial explanation for this situation comes from in silico modeling , which predicts that the docking site for p25 on GSK3β is the O15169 -binding domain ; because of this , p25 inhibits the formation of the GSK3β/ O15169 / P25054 destruction complex , thus preventing GSK3β from binding to and phosphorylating β-catenin . Coexpression of GSK3β and p25 in cultured neurons results in a neurodegeneration phenotype that exceeds that observed with Q00535 and p25 . When p25 is transfected alone , the resulting neuronal damage is blocked more effectively with a specific siRNA against Gsk3β than with one against Cdk5 . We propose that the effects of p25 , although normally attributed to activate Q00535 , may be mediated in part by elevated GSK3β activity . Caspase activation may be associated with Mycobacterium avium pathogenicity . BACKGROUND : Mycobacterium avium causes disseminated infection in immunocompromised patients and triggers a process resembling Crohn 's disease in goats . Colony morphotypes predict pathogenicity . Smooth-transparent ( SmT ) morphotypes are more virulent and induce less interleukin ( IL ) -1beta and Q14116 production than avirulent smooth-domed ( SmD ) morphotypes . Caspases are essential for IL-1beta and Q14116 production . METHODS : Caspase activation was examined in human monocytes after M. avium infection . RESULTS : Fresh monocytes constitutively expressed caspase-1 mRNA and pro-caspase-1 . The M. avium infection increased monocyte caspase-1 mRNA expression . Furthermore , SmD-infected monocytes expressed 2.3-fold higher levels ( P < 0.05 , n = 3 ) of activated caspases than SmT-infected monocytes . P29466 inhibition significantly reduced IL-1beta production by SmT- and SmD-infected monocytes ( P < 0.05 , n = 4 ) . P42574 inhibition inhibited IL-1beta production 43.5 % +/- 8.0 % ( P < 0.02 , n = 4 ) by SmD-infected but not SmT-infected monocytes . CONCLUSIONS : Decreased mature IL-1beta release by SmT-infected monocytes may reflect selective induction of caspase-1 activity but not caspase-3 . Differential caspase expression in monocytes after infection may contribute to M. avium pathogenicity in humans . 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 . 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 . 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 . Endotoxemic acute renal failure is attenuated in caspase-1-deficient mice . P29466 -deficient ( -/- ) mice are protected against sepsis-induced hypotension and mortality . We investigated the role of caspase-1 and its associated cytokines in a nonhypotensive model of endotoxemic acute renal failure ( Q8N726 ) . Mice were injected intraperitoneally with 2.5 mg of LPS that induces endotoxemic Q8N726 . On immunoblot analysis of whole kidney , there was an increase in caspase-1 protein in LPS-treated mice compared with vehicle-treated controls . In LPS-treated mice , the glomerular filtration rate ( Q92565 ) was significantly higher in caspase-1 -/- vs. wild-type mice at 16 and 36 h after LPS . To determine the mechanism of this protection , the caspase-1-activated cytokines IL-1beta and Q14116 were investigated . IL-1beta and Q14116 protein were significantly increased in the kidneys of LPS- vs. vehicle-treated mice . To determine the role of these cytokines , mice were treated with recombinant IL-1 receptor antagonist ( IL-1Ra ) or Q14116 -neutralizing antiserum . In LPS-treated mice , Q92565 was not different in IL-1Ra-treated or Q14116 -neutralizing antiserum-treated or combination therapy ( IL-1Ra plus Q14116 -neutralizing antiserum-treated ) compared with control mice . In addition , tubular cell apoptosis , neutrophil infiltration , myeloperoxidase activity , caspase-3 activity , and calpain activity were not different between wild-type and caspase-1 -/- mice with endotoxemic Q8N726 . In LPS- vs. vehicle-treated wild-type mice , renal IL-1alpha was significantly increased . In both LPS- and vehicle-treated caspase-1 -/- mice , renal IL-1alpha was very low . In summary , caspase-1 -/- mice are functionally protected against endotoxemic Q8N726 . Neutralization of IL-1beta and Q14116 is not functionally protective . The role of the intracellular proinflammatory cytokine IL-1alpha in endotoxemic Q8N726 merits further study . DB04690 induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani . The parasites of the order kinetoplastidae including Leishmania spp. emerge from most ancient phylogenic branches of unicellular eukaryotic lineages . In their life cycle , topoisomerase I plays a significant role in carrying out vital cellular processes . DB04690 ( CPT ) , an inhibitor of P11387 , induces programmed cell death ( P61457 ) both in the amastigotes and promastigotes form of L. donovani parasites . CPT-induced cellular dysfunction in L. donovani promastigotes is characterized by several cytoplasmic and nuclear features of apoptosis . CPT inhibits cellular respiration that results in mitochondrial hyperpolarization taking place by oligomycin-sensitive F0-F1 ATPase-like protein in leishmanial cells . During the early phase of activation , there is an increase in reactive oxygen species ( ROS ) inside cells , which causes subsequent elevation in the level of lipid peroxidation and decrease in reducing equivalents like DB00143 . Endogenous ROS formation and lipid peroxidation cause eventual loss of mitochondrial membrane potential . Furthermore , cytochrome c is released into the cytosol in a manner independent of involvement of CED3/ P42574 group of proteases and unlike mammalian cells it is insensitive to cyclosporin A . These events are followed by activation of both CED3/ P42574 and ICE group of proteases in P61457 of Leishmania . Taken together , our study indicates that different biochemical events leading to apoptosis in leishmanial cells provide information that could be exploited to develop newer potential therapeutic targets . 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 . 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 . Regional selective neuronal degeneration after protein phosphatase inhibition in hippocampal slice cultures : evidence for a Q96HU1 kinase-dependent mechanism . The regional selectivity and mechanisms underlying the toxicity of the serine/threonine protein phosphatase inhibitor okadaic acid ( OA ) were investigated in hippocampal slice cultures . Image analysis of propidium iodide-labeled cultures revealed that okadaic acid caused a dose- and time-dependent injury to hippocampal neurons . Pyramidal cells in the P07451 region and granule cells in the dentate gyrus were much more sensitive to okadaic acid than the pyramidal cells in the P00915 region . Electron microscopy revealed ultrastructural changes in the pyramidal cells that were not consistent with an apoptotic process . Treatment with okadaic acid led to a rapid and sustained tyrosine phosphorylation of the mitogen-activated protein kinases P27361 and P28482 ( Q8TCB0 /42(mapk) ) . The phosphorylation was markedly reduced after treatment of the cultures with the microbial alkaloid DB02152 ( a nonselective protein kinase inhibitor ) or the Q96HU1 kinase kinase ( Q02750 /2 ) inhibitor PD98059 . DB02152 and PD98059 also ameliorated the okadaic acid-induced cell death . Inhibitors of protein kinase C , Ca2+/calmodulin-dependent protein kinase II , or tyrosine kinase were ineffective . These results indicate that sustained activation of the Q96HU1 kinase pathway , as seen after e.g. , ischemia , may selectively harm specific subsets of neurons . The susceptibility to Q96HU1 kinase activation of the P07451 pyramidal cells and dentate granule cells may provide insight into the observed relationship between cerebral ischemia and dementia in Alzheimer 's disease . The anti-androgen drug dutasteride renders triple negative breast cancer cells more sensitive to chemotherapy via inhibition of HIF-1α-/ P15692 -signaling . BACKGROUND : Triple negative breast cancer ( TNBC ) is characterized by lack of expression of both estrogen and progesterone receptor as well as lack of amplification of P04626 . Patients with TNBC carry an unfavorable prognosis compared to other breast cancer subtypes given that endocrine or P04626 targeted therapies are not effective , rendering chemotherapy the sole effective treatment option to date . Therefore , there is a high demand for additional novel treatment options . FINDINGS : We previously published a list of genes showing both higher gene expression rates in TNBC and , in addition , are known to encode targets of non-oncologic drugs . P18405 , which encodes the type-1 isoform of the steroid-5alpha-reductase , which is involved in androgen metabolism , was found to be one of these genes . DB01126 is a dual blocker of both the type-1 and type-2 isoform of P18405 and is indicated in the treatment of benign prostate hyperplasia . Treatment of TNBC cell lines with dutasteride was associated with a dose-dependent decrease in cell viability , altered protein expression of P15692 and HIF-1α and increased chemosensitivity . CONCLUSION : Our results demonstrate that the P18405 -corresponding anti-androgenic drug dutasteride might act as a combinatorial therapeutic option besides standard chemotherapy in highly aggressive TNBC . Potent antitumor efficacy of interleukin-18 delivered by conditionally replicative adenovirus vector in renal cell carcinoma-bearing nude mice via inhibition of angiogenesis . It has been demonstrated that interleukin 18 ( Q14116 ) exerts antitumor activity . In this study , we investigated whether oncolytic adenovirus-mediated gene transfer of Q14116 could induce strong antitumor activity . A tumor-selective replicating adenovirus expressing Q14116 ( ZD55- Q14116 ) was constructed by insertion of an Q14116 expression cassette into the ZD55 vector , which is based on deletion of the adenoviral E1B 55-kDa gene . ZD55- Q14116 could express substantially more Q14116 than Ad- Q14116 because of replication of the vector . It has been shown that ZD55- Q14116 exerted a strong cytopathic effect and significant apoptosis in renal cell carcinoma . ZD55- Q14116 significantly decreased P15692 and P28906 expression in the tumor cells . Treatment of established tumors with ZD55- Q14116 showed much stronger antitumor activity than that induced by ZD55-EGFP or Ad- Q14116 . These data indicated that oncolytic adenovirus expressing Q14116 could exert potential antitumor activity via inhibition of angiogenesis and offer a novel approach to cancer therapy . 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 . 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 .

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interacts_with DB00215?

multiple_choice

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 . Immunotoxicity of aminocarb . III . Exposure route-dependent immunomodulation by aminocarb in mice . Aminocarb , a phenylsubstituted methylcarbamate pesticide ( 4-dimethylamino-3-methyl-N-carbamate ; matacil ) , previously suspected of a relatively low immunotoxic potential , was administered by four different exposure routes to C57BL/6 mice . A single sublethal exposure by oral and dermal routes stimulated humoral immune response at a relatively low dose ; 1/256 LD50 of aminocarb . Intraperitoneal ( i.p. ) injection decreased the humoral P27918 response , whereas inhalation of aminocarb had no marked effect on peripheral immune status in exposed animals . Thus , i.p. exposure resulted in higher immunotoxicity over oral administration of aminocarb . Similarly , marked route-related exposure differences in immunomodulatory effects of aminocarb were noted for mitogenic stimulation of spleen lymphocytes and mixed lymphocyte response . Other indices , such as delayed type hypersensitivity ( DTH ) and production of interleukin-2 ( P60568 ) were unchanged . Interestingly , expression of major histocompatibility complex ( MHC ) class II by purified , lipopolysaccharide ( LPS ) -stimulated B cells increased equally after i.p. and oral exposures to aminocarb . Overall , a weak immunosuppressive potential of aminocarb was concluded , which was possibly due to indirect interaction of the pesticide with the immune system . However , aminocarb may represent an autoimmunity-inducing toxic . The c- DB00134 receptor tyrosine kinase inhibitor DB05216 radiosensitizes glioblastoma cells . PURPOSE : Glioblastoma multiforme ( GBM ) is resistant to current cytotoxic therapies , in part because of enhanced DNA repair . Activation of the receptor tyrosine kinase c- DB00134 has been shown to protect cancer cells from DNA damage . We hypothesized that inhibiting c- DB00134 would decrease this protection and thus sensitize resistant tumor cells to the effects of radiation therapy . MATERIALS AND METHODS : Eight human GBM cell lines were screened for radiosensitivity to the small-molecule c- DB00134 inhibitor DB05216 with colony-count assays . Double-strand ( ds ) DNA breaks was quantified by using antibodies to gamma P16104 . Western blotting demonstrate expression of Q06609 , glycogen synthase kinase ( GSK ) -3beta , and other proteins . A murine xenograft tumor flank model was used for in vivo radiosensitization studies . RESULTS : DB05216 reduced c- DB00134 phosphorylation and enhanced radiation-induced cell kill by 0.4 logs in SF767 cells . Cells pretreated with DB05216 had more ds DNA damage than cells treated with radiation alone . Mechanistically , DB05216 was shown to inhibit dsDNA break repair and increase apoptosis . DB05216 influences various survival and DNA repair related proteins such as pAKT , Q06609 and GSK3beta . In vivo , the addition of DB05216 to radiation resulted in a tumor-growth-delay enhancement ratio of 2.9 over radiation alone and extended survival time . CONCLUSIONS : GBM is a disease site where radiation is often used to address both macroscopic and microscopic disease . Despite attempts at dose escalation outcomes remain poor . DB05216 , a potent small-molecule tyrosine kinase inhibitor of c- DB00134 , radiosensitized several GBM cell lines both in vitro and in vivo , and may help to improve outcomes for patients with GBM . Administration of adenosine diphosphate-ribosyl transferase antagonist allows in vivo control of anti-dinitrophenyl response . DB03073 ( 3MB ) is one of a series of chemical inhibitors of the nuclear enzyme adenosine diphosphate ( ADP ) -ribosyl transferase ( P09874 ) , which has been shown to inhibit cell differentiation in vitro , but has no effect on differentiation independent proliferation . Treatment of mice with an optimal concentration of 3MB ( 20 mg/kg body weight ) at or 1 day after dinitrophenyl-keyhole limpet haemocyanin ( DNP-KLH ) immunisation reduced anti-DNP plaque-forming cell ( P27918 ) numbers to less than 10 % of those of control animals . The period for maximum P27918 suppression showed a narrow time window relative to immunisation , suggesting that in vivo , as in vitro , 3 MB was acting only on those lymphocytes differentiating in response to antigen . Experimental findings showed that it was possible to select for P27918 derived from different populations of DNP-responsive lymphocytes by adjusting the time of 3MB treatment relative to immunisation . When 3MB was used with antigen priming , the residual P27918 showed a lower average affinity than P27918 in mice treated with 3MB 3 days after priming , suggesting a differential selection of those lymphocytes responding either ' early ' or ' late ' in the primary immune response . Displacement of Q01826 -bound histone deacetylase 1 corepressor by the human immunodeficiency virus type 1 transactivator induces expression of interleukin-2 and its receptor in T cells . One hallmark of human immunodeficiency virus type 1 ( HIV-1 ) infection is the dysregulation of cytokine gene expression in T cells . Transfection of T cells with human T-cell leukemia type 1 or 2 transactivator results in the induction of the T-cell-restricted cytokine interleukin-2 ( P60568 ) and its receptor ( IL-2Ralpha ) . However , no T-cell-specific factor(s) has been directly linked with the regulation of P60568 and IL-2Ralpha transcription by influencing the promoter activity . Thymocytes from Q01826 ( special AT-rich sequence binding protein 1 ) knockout mice have been shown to ectopically express IL-2Ralpha , suggesting involvement of Q01826 in its negative regulation . Here we show that Q01826 , a T-cell-specific global gene regulator , binds to the promoters of human P60568 and IL-2Ralpha and recruits histone deacetylase 1 ( Q13547 ) in vivo . Q01826 also interacts with Tat in HIV-1-infected T cells . The functional interaction between HIV-1 Tat and Q01826 requires its PDZ-like domain , and the binding of the Q13547 corepressor occurs through the same . Furthermore , Tat competitively displaces Q13547 that is bound to Q01826 , leading to increased acetylation of the promoters in vivo . Transduction with Q01826 interaction-deficient soluble Tat ( Tat 40-72 ) and reporter assays using a transactivation-negative mutant ( C22G ) of Tat unequivocally demonstrated that the displacement of Q13547 itself is sufficient for derepression of these promoters in vivo . These results suggest a novel mechanism by which HIV-1 Tat might overcome Q01826 -mediated repression in T cells . 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 . Significant reduction of granulomas in Nrf2-deficient mice infected with Mycobacterium tuberculosis . OBJECTIVE : We have reported previously that mice deficient in nuclear erythroid 2 P29466 -related factor 2 ( Nrf2 ) , which regulates the expression of antioxidant and detoxification genes , showed significant susceptibility to airway inflammatory responses when exposed to diesel exhaust particles for eight weeks . As disruption of Nrf2 promotes immune cells that stimulate Th2-like immunoresponsiveness , Nrf2-deficient mice may be resistant to M. tuberculosis infection . SETTING : Nrf2-deficient mice were infected with M. tuberculosis aerially , and the size of their granulomas and cytokine mRNA expression were compared with those of wild-type mice . RESULTS : Significant reduction of granuloma formation and tubercle bacilli in granulomas was noted in the deficient mice 27 weeks after infection , concurrently with higher expression of P60568 and P35225 mRNA . CONCLUSION : It is concluded that Nrf2 inversely regulates M. tuberculosis-induced granuloma development at the late stage . Inactivation of caspase-1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin-1 ( IL-1 ) -beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL-1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL-1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE/caspase-1 ) or through caspase-1 gene deletion . METHODS : P29466 was selectively blocked by using pralnacasan or DB05507 . IL-1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL-1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 inhibition reduced the release of IL-1beta in organotypic slices exposed to LPS+ DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 ( intraperitoneal , 25-200 mg/kg ) to rats blocked seizure-induced production of IL-1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase-1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase-1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL-1beta . 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 . 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 . Gamma-aminobutric acid and glutamate decarboxylas ( l-glutamate 1-carboxy-lyase e.c. 4.1.1.15 ) in the nervous system of the cockroach , periplaneta americana.i.regional distribution and properties of the enzyme . Both the central and peripheral nervous system of the cockroach Periplaneta americana contain gamma-aminobutyric acid ( GABA ) and glutamate decarboxylase ( Q99259 ) . In the central ganglia of the cockroach , an average of more that 60 mumoles of GABA are formed from glutamate ( DB00142 ) per gram wet weight of tissue per hour . This activity level of the Q99259 apoenzyme is considerably higher than that found in the central nervous system of crustaceans , amphibians , avians and mammals but is similar to that reported for nervous system tissues from other insect species . A comparison of properties of the crude cockroach enzyme with Q99259 from crustacean and mammalian origin revealed both similarities and differences : whereas crude cockroach Q99259 has cofactor requirements and an affinity for DB00142 substrate ( Km 2.8 X 10-2 ) which are similar to Q99259 from lobster and mouse , it is uniquely inhibited by both Cl-and by GABA . The Q99259 from cockroach nervous tissues has two apparent pH optima of which the lower one is preferentially inhibited by a compound which is found in the nerve sheath and the fat body tissue adjacent to ganglia and axons . Intranasal immunization against herpes simplex virus infection by using a recombinant glycoprotein D fused with immunomodulating proteins , the B subunit of Escherichia coli heat-labile enterotoxin and interleukin-2 . To establish a novel strategy of mucosal immunization against herpes simplex virus type 1 ( HSV-1 ) infection , we studied the immune responses elicited by intranasal immunization with several forms of a recombinant glycoprotein D ( gD ) of HSV-1 . A truncated gD ( t-gD ) co-administered with heat-labile enterotoxin B subunit ( Q06643 ) from Escherichia coli induced both a mucosal immune response involving secretion of anti-gD IgA and serum IgG production . The levels of these responses are comparable to those in mice which have recovered from intranasal HSV-1 infections . The fusion protein ( t-gD- Q06643 ) , consisting of t-gD and Q06643 , induced the responses more efficiently than did co-administration of t-gD and Q06643 , although GM1 ganglioside binding activity was significantly reduced in t-gD- Q06643 . We found that another fusion protein , consisting of t-gD and human interleukin-2 ( t-gD- P60568 ) , also elicited antibody responses comparable to those induced by t-gD- Q06643 . Immunity acquired by intranasal immunization with t-gD- P60568 protected mice from intraperitoneal HSV-1 infections , whereas t-gD- Q06643 or t-gD alone failed to provide protection against infection . Even in a mouse strain that responded highly to subcutaneously administered gD , intranasally administered t-gD did not elicit antibody responses . The lack of response to gD was clearly abrogated by co-administration with P60568 , and administration of t-gD- P60568 induced an excellent level of antibody responses in this strain . These results suggest that the P60568 fusion strategy yields a new type of mucosal immunization , the mechanism of which differs from that speculated for the mucosal adjuvant activity of Q06643 . 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 . 5-Lipoxygenase-activating protein ( P20292 ) inhibitors . Part 4 : development of 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid ( AM803 ) , a potent , oral , once daily P20292 inhibitor . The potent P09917 -activating protein ( P20292 ) inhibitor 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid 11cc is described ( AM803 , now GSK2190915 ) . Building upon DB05225 ( 1 ) ( Hutchinson et al. J. Med Chem.2009 , 52 , 5803-5815 ; Stock et al. Bioorg. Med. Chem. Lett. 2010 , 20 , 213-217 ; Stock et al. Bioorg. Med. Chem. Lett.2010 , 20 , 4598-4601 ) , SAR studies centering around the pyridine moiety led to the discovery of compounds that exhibit significantly increased potency in a human whole blood assay measuring Q06643 (4) inhibition with longer drug preincubation times ( 15 min vs 5 h ) . Further studies identified 11cc with a potency of 2.9 nM in P20292 binding , an IC(50) of 76 nM for inhibition of Q06643 (4) in human blood ( 5 h incubation ) and excellent preclinical toxicology and pharmacokinetics in rat and dog . 11cc also demonstrated an extended pharmacodynamic effect in a rodent bronchoalveolar lavage ( BAL ) model . This compound has successfully completed phase 1 clinical studies in healthy volunteers and is currently undergoing phase 2 trials in asthmatic patients . Biological effects of alpha particle radiation exposure on human monocytic cells . Radon ( (222)Rn ) gas produces decay progeny that emits high energy alpha (α)-particles . Epidemiological studies have shown that exposure to (222)Rn is linked with elevated risk of developing lung cancer , however clear mechanisms leading to such effects have not been delineated . Cytokines play a critical role in inflammation and their dysregulated production often contributes to disease pathogenesis . In this study , Bio-plex multiplex technology was employed to investigate modulations of 27 pro-inflammatory cytokines following exposure of human monocytic cells to 1.5 Gy of α-particle radiation . Concurrently , DNA damage was assessed by examining the formation of phosphorylated H2A histone family X ( γ- P16104 ) sites . Of the 27 cytokines assessed , 4 cytokines were shown to be statistically downregulated by ∼2 fold relative to the untreated controls and included the interleukin ( IL ) family of proteins ( P60568 , P40933 and Q16552 ) and macrophage inflammatory protein 1 beta ( MIP-1b ) . Interferon-inducible protein-12 ( IP-12 ) , vascular endothelial growth factor and regulated on activation normal T cell expressed and secreted ( RANTES ) were shown to be high expressors and upregulated . Cells irradiated with α-particles ranging from 0.27 to 2.14 Gy showed statistically significant , dose-dependant increases in γ- P16104 formation . These data suggest that α-particle radiation causes dysregulation in the production of a number of pro-inflammatory cytokines and results in significant DNA damage . 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 . 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 . Presence of diabetes-inhibiting , glutamic acid decarboxylase-specific , P22301 -dependent , regulatory T cells in naive nonobese diabetic mice . Immunization of NOD mice with autoantigens such as glutamic acid decarboxylase ( Q99259 ) 221-235 peptide ( p221 ) can induce Ag-specific P01730 (+) T regulatory ( Tr ) cells . However , it is unclear whether these Tr cells acquire their regulatory capacity due to immunization or whether they are constitutively harbored in unimmunized naive mice . To address this question , we used an I-Ag7 tetramer to isolate p221-specific T cells from naive NOD mice ( N221(+) cells ) after peptide-specific in vitro expansion . The N221(+) T cells produced P01579 and P22301 , but very little P05112 , in response to p221 stimulation . These T cells could function as regulatory cells and inhibit in vitro proliferation of diabetogenic BDC2.5 cells . This suppressive activity was cell contact-independent and was abrogated by Abs to P22301 or IL-10R . Interestingly , P60568 produced by other T cells present in the cell culture induced unactivated N221(+) T cells to exhibit regulatory activities involving production of P22301 . In vivo , N221(+) cells inhibited diabetes development when cotransferred with NOD splenocytes into NOD/scid recipients . Together , these results demonstrate that p221-specific P22301 -dependent Tr cells , including Tr type 1 cells , are present in naive NOD mice . The use of spontaneously arising populations of Q99259 peptide-specific Tr cells may represent a promising immunotherapeutic approach for preventing type 1 diabetes . Diabetes mellitus in cancer patients treated with combination interleukin 2 and alpha-interferon . Diabetes mellitus is thought to be an autoimmune disease caused by destruction of beta cells in pancreatic islets . P01308 resistance in the peripheral tissues may also play a role . Both interleukin 2 ( P60568 ) and alpha interferon can enhance immune function by stimulating formation of cytolytic T cells and/or antigen expression on both normal and tumor cells . This report describes three patients with advanced malignancy who were treated with combination P60568 and alpha interferon who had the onset or worsening of diabetes mellitus . One patient died as a result . There is evidence that interferon can increase insulin resistance and it is likely that both agents can initiate or enhance an ongoing autoimmune process . Physicians using this combination of drugs should be aware of this potential serious toxicity . [ The importance of determination of interleukin-10 in the blood of patients with systemic lupus erythematosus ] . BACKGROUND : Hitherto , not very numerous investigations provided so far only few often controversial findings on the importance of interleukin-10 ( P22301 ) in systemic lupus erythematosus ( SLE ) . The objective of the present investigation was to assess whether there exist practically applicable relations between the serum level of P22301 , clinical and laboratory indicators of activity of the disease and serum levels of selected cytokines or their soluble receptors . METHODS AND RESULTS : The authors analyzed a group of 23 patients with SLE ( 23 women and 1 man , median age 37 years ) . P22301 and other cytokines were examined by the ELISA method , the clinical activity of the disease was evaluated by the ECLAM system ( European Consensus Lupus Activity Measurement ) . Elevated P22301 values ( > 5 pg/ml ) were assessed in 10 ( 43 % ) patients . Correlation analysis ( Pearson 's test , p < 0.05 ) revealed statistically significant relations between P22301 levels and the activity of the disease , values of antibody levels against dsDNA and levels of the soluble receptor P60568 ( sIL-2R ) in serum . Conversely , no relationship was revealed between values of P22301 and values of P01024 and C4 complement components , IL-1 , P60568 , P05231 , sIL-6R , P01375 , sTNFR-alpha and P27352 -gamma . CONCLUSIONS : Elevated P22301 serum levels in patients with SLE did not have , with the exception of the index of clinical activity of the disease , antibodies against dsDNA and sIL-2R any statistically significant relations to laboratory indicators of disease activity and levels of selected cytokines and their soluble receptors .

MH_train_1558

MH_train_1558

MH_train_1558

interacts_with DB00559?

multiple_choice

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 . In vitro effects and ex vivo binding of an P00533 -specific immunotoxin on rhabdomyosarcoma cells . PURPOSE : Rhabdomyosarcoma ( RMS ) is a rare and aggressive soft tissue sarcoma with limited treatment options and a high failure rate during standard therapy . New therapeutic strategies based on targeted immunotherapy are therefore much in demand . The epidermal growth factor receptor ( P00533 ) has all the characteristics of an ideal target . It is overexpressed in up to 80 % of embryonal RMS and up to 50 % of alveolar RMS tumors . We therefore tested the activity of the P00533 -specific recombinant immunotoxin ( IT ) 425(scFv)- P25101 ' against P00533 (+) RMS cells in vitro and ex vivo . METHODS : We tested the specific binding and internalization behavior of 425(scFv)- P25101 ' in RMS cell lines in vitro by flow cytometry , compared to the corresponding imaging probe 425(scFv)- P60880 monitored by live cell imaging . The cytotoxic activity of 425(scFv)- P25101 ' was tested using cell viability and apoptosis assays . Specific binding of the IT was confirmed on formalin-fixed paraffin-embedded tissue samples from two RMS patients . RESULTS : We confirmed the specific binding of 425(scFv)- P25101 ' to RMS cells in vitro and ex vivo . Both the IT and the corresponding imaging probe were rapidly internalized . The IT killed P00533 (+) RMS cells in a dose-dependent manner , while showing no effect against control cells . It showed specific apoptotic activity against one selected RMS cell line . CONCLUSIONS : This is the first study showing the promising therapeutic potential of a recombinant , P00533 -targeting , P25101 '-based IT on RMS cells . We confirmed the selective killing with IC50 values of up to 50 pM , and immunohistochemical staining confirmed the specific ex vivo binding to primary RMS material . Effect of cytokines and ovarian steroids on equine endometrial function : an in vitro study . Regulation of immune-endocrine interactions in the equine endometrium is not fully understood . The aims of the present study were to : ( 1 ) investigate the presence of tumour necrosis factor alpha ( P01375 ) , interferon gamma ( P01579 ) , P48023 ( P48023 ) and their receptors in the mare endometrium throughout the oestrous cycle ; and ( 2 ) assess endometrial secretory function ( prostaglandins ) , angiogenic activity and cell viability in response to P01375 , oestradiol ( E2 ) , progesterone ( P4 ) and oxytocin ( P01178 ) . Transcription of P01375 and P48023 mRNA increased during the early and late luteal phase ( LP ) , whereas P01579 mRNA increased in late LP . Transcription of the mRNA of both P01375 receptors was highest in the mid-LP . All cytokines and receptors were expressed in surface and glandular epithelium , as well as in the stroma . Expression of P01375 and its receptor P19438 increased during the follicular phase ( FP ) and mid-LP . P01579 was expressed in the mid-LP , whereas its receptor IFNR1 was expressed in the in mid- and late LP . The highest expression of P48023 and FAS occurred during the late LP . P01178 increased the secretion of prostaglandin ( PG ) E2 and PGF2α in the FP and mid-LP . In the mid-LP , E2 and P4+E2 stimulated PGF2α secretion , whereas P01375 and P4 increased cell viability . All treatments , with the exception of P4 , increased nitric oxide and angiogenic activity in both phases . The coordinated action of cytokines and ovarian hormones may regulate secretory , angiogenic and proliferative functions in the equine endometrium . 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 . Characterization and autoradiographic localization of histamine H1 receptors in human nasal turbinates . To examine the localization of histamine H1 receptors ( P35367 ) in human nasal mucosa , the autoradiographic distribution of P35367 was studied in human nasal inferior turbinates . Cryostat sections were incubated with various concentration of [3H] DB06691 in saturation-binding studies and with 1 nmol/L of [3H] DB06691 for autoradiography . Nonspecific binding was determined by adding 2 mumol/L of DB06691 . Scatchard analysis demonstrated high-affinity binding sites with a maximum binding capacity of P35367 of 193 +/- 46 fmol/mg of protein , and dissociation constant was 0.6 +/- 0.1 nmol/L . Autoradiograms indicated P35367 exist exclusively on the endothelium of vessels . No specific labeling could be observed in the submucosal glands or epithelium . These results extend and support our previous finding that histamine directly causes vascular permeability through P35367 and stimulates nasal glandular secretion indirectly through reflexes . 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 . Dual endothelin receptor antagonism prevents remodeling of resistance arteries in diabetes . Vascular remodeling , characterized by extracellular matrix deposition and increased media-to-lumen ( M/L ) ratio , contributes to the development of microvascular complications in diabetes . We have previously shown in type 2 diabetic Goto-Kakizaki ( GK ) rats that selective P25101 receptor blockade prevents medial thickening of mesenteric arteries via regulation of matrix metalloproteases ( MMP ) , whereas selective ETB receptor blockade augments this thickening . The goal of this study was to determine the effect of combined P25101 and ETB receptor blockade on resistance vessel remodeling . Vessel structure , MMP activity , and extracellular matrix proteins were assessed in control Wistar and diabetic GK rats treated with vehicle or DB00559 ( 100 mg/kg per day ) for 4 weeks ( n = 7-9 per group ) . DB00559 completely prevented the increase in M/L ratio and P08253 activity in diabetes but paradoxically increased M/L ratio and MMP activation in control animals . Collagenase ( P45452 ) activity and protein levels were significantly decreased in diabetes . Accordingly , collagen deposition was augmented in GK rats . Dual ET receptor antagonism improved enzyme activity and normalized P45452 levels in diabetic animals but blunted P45452 activity in control animals . In summary , current findings suggest that diabetes-mediated remodeling of resistance arteries is prevented by dual blockade of P25101 and ETB receptors and that the relative role of ET receptors in the regulation of vascular structure differs in the control and disease states . 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 . Structure-activity relationship studies of CNS agents -- XVII . Spiro[piperidine-4',1-(1,2,3,4-tetrahydro-beta-carboline)] as a probe defining the extended topographic model of P08908 receptors . Spiro[piperidine-4',1-(1,2,3,4-tetrahydro-beta-carboline)] ( 10 ) , its derivatives 11-15 and its analogs 16 and 17 were examined as ligands of serotonin P08908 receptors . It was shown that compounds 12 and 14 had essentially the same P08908 affinity as 1-phenylpiperazine and its rigid analog 7 , whereas there were substantial differences in the steric arrangement of their crucial pharmacophores , i.e. aromatic and protonation centers . On the basis of the existing models and using the DB04829 structure as a template , a new , extended three-point topographic model of P08908 receptors has been proposed . 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 % . DB06693 enhances osteogenesis in murine embryonic stem cells . Embryonic stem ( ES ) cells have the capacity to differentiate into various cell types in vitro . In this study , we show that retinoic acid is important for the commitment of ES cells into osteoblasts . Culturing retinoic acid treated ES cells in the presence of the osteogenic supplements ascorbic acid and beta-glycerophosphate resulted in the expression of several osteoblast marker genes , osteocalcin , alkaline phosphatase , and osteopontin . However , there was only a slight amount of mineralized matrix secretion . Addition of bone morphogenic protein-2 or compactin , a drug of the statin family of P04035 inhibitors , resulted in a greatly enhanced formation of bone nodules . DB06693 did not modify the expression of osteogenic markers , but at the late stage of differentiation promoted an increase in P12643 expression . These results establish ES-cell derived osteogenesis as an effective model system to study the molecular mechanisms by which the statin compactin promotes osteoblastic differentiation and bone nodule formation . 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 . 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 . P15502 -based nanoparticles for delivery of bone morphogenetic proteins . Biocompatibility , stability , and efficiency remain among the major goals in the development of new drug delivery systems . Herein we describe a facile method for encapsulation of Bone Morphogenetic Protein-2 ( P12643 ) at high concentrations and in mild conditions by exploiting the thermo-responsiveness of an elastin-like polymer . Chemically synthesized poly(VPAVG) demonstrates excellent biocompatibility , stability at room temperature , and allows the use of mild conditions and harmless solvents , such as water , for its production . Here , we describe how to use a recombinantly produced poly(VPAVG) polymer to entrap P12643 with a high encapsulation efficiency and the methods for in vitro bioactivity assays . Truncation of P60880 reduces the stimulatory action of DB02527 on rapid exocytosis in insulin-secreting cells . Synaptosomal protein of 25 kDa ( P60880 ) is important for Ca(2+)-dependent fusion of large dense core vesicles ( LDCVs ) in insulin-secreting cells . Exocytosis is further enhanced by DB02527 -increasing agents such as glucagon-like peptide-1 ( P0C6A0 ) , and this augmentation includes interaction with both PKA and Q8WZA2 . To investigate the coupling between P60880 - and DB02527 -dependent stimulation of insulin exocytosis , we have used capacitance measurements , protein-binding assays , and Western blot analysis . In insulin-secreting P01308 -1 cells overexpressing wild-type P60880 ( P60880 (WT) ) , rapid exocytosis was stimulated more than threefold by DB02527 , similar to the situation in nontransfected cells . However , DB02527 failed to potentiate rapid exocytosis in P01308 -1 cells overexpressing a truncated form of P60880 ( P60880 (1-197) ) or Botulinum neurotoxin A ( DB00083 ) . Close dissection of the exocytotic response revealed that the inability of DB02527 to stimulate exocytosis in the presence of a truncated P60880 was confined to the release of primed LDCVs within the readily releasable pool , especially from the immediately releasable pool , whereas DB02527 enhanced mobilization of granules from the reserve pool in both P60880 (1-197) ( P < 0.01 ) and P60880 (WT) ( P < 0.05 ) cells . This was supported by hormone release measurements . Augmentation of the immediately releasable pool by DB02527 has been suggested to act through the Q8WZA2 -dependent , PKA-independent pathway . Indeed , we were able to verify an interaction between P60880 with both Q8WZA2 and Q9UQ26 , two proteins involved in the PKA-independent pathway . Thus we hypothesize that P60880 is a necessary partner in the complex mediating DB02527 -enhanced rapid exocytosis in insulin-secreting cells . DB00107 in Brattleboro rats : increased synthesis is contrasted by blunted intrahypothalamic release from supraoptic nucleus neurones . Adult male Brattleboro rats were used to investigate the impact of the congenital absence of vasopressin on the release pattern of oxytocin ( P01178 ) within the hypothalamic supraoptic nucleus ( P18583 ) in response to a 10-min forced swimming session and osmotic stimulation . Both immunohistochemical and in situ hybridisation data suggest that vasopressin-deficient animals have more oxytocin-synthesising neurones in the P18583 than homozygous wild-type controls . Unexpectedly , both forced swimming and peripheral osmotic stimulation resulted in a blunted release profile of oxytocin within the P18583 of vasopressin-deficient rats compared to controls . A similar intranuclear P01178 response to direct osmotic stimulation of the P18583 by retrodialysis with hypertonic Ringer 's solution in both genotypes confirmed the capability of P18583 neurones to locally release oxytocin in vasopressin-deficient rats , indicating an altered processing of information originating from multisynaptic inputs rather than a deficit in release capacity . Taken together with data obtained in previous studies , the present findings provide evidence suggesting that autocrine and paracrine signalling of magnocellular neurones differs within the paraventricular nucleus and the P18583 . Thus , significant alterations in intra- P18583 oxytocin mRNA levels can not easily be extrapolated to intranuclear release profiles and the local signal intensity of this neuropeptide after physiological stimulation . Respective role of humoral factors and blood pressure in aortic remodeling of DOCA hypertensive rats . Hypertension results in increased thickness and stiffness of large artery walls . The goal of our study was to assess the respective roles of humoral factors such as Ang II , endothelin and blood pressure in these aortic modifications . For this purpose , uninephrectomized rats received DOCA and high salt diet , and when hypertension was installed , they were treated for 5 weeks with either a long-acting calcium antagonist , mibefradil ( 30 mg/kg/day ) , an P12821 inhibitor , enalapril ( 3 mg/kg/day ) , or a mixed P25101 and ETB endothelin receptor antagonist , DB00559 ( 100 mg/kg/day ) . A group of hypertensive rats was left untreated and a sham-operated group of normotensive rats was used for control . At the end of treatment , aortic medial thickness and elastin as well as collagen were evaluated by quantitative morphometry . DOCA-salt hypertensive rats exhibited a marked increase in medial thickness associated with no change in absolute content in extracellular matrix . P15502 relative density decreased in DOCA rats . Enalapril had no effect on arterial pressure . DB00559 decreased slightly ( by 12 mm Hg ) , but not significantly , blood pressure . None of these drugs had an effect on medial thickness suggesting that in DOCA hypertensive rats neither Ang II nor endothelin play a significant role in the remodeling of the aorta . In contrast , mibefradil almost normalized arterial pressure , prevented medial hypertrophy and increased elastin density . Further studies are required in order to assess if this effect is directly linked to the blood pressure decrease or to another mechanism related to the calcium antagonistic property of mibefradil . Nonsteroidal anti-inflammatory drugs and inflammatory bowel disease : current perspectives . Mechanisms underlying the gastric toxicity of nonsteroidal anti-inflammatory drugs ( NSAIDs ) have been extensively investigated , whereas those leading to intestinal damage are not completely understood . Several hypotheses have been put forward on the pathophysiology of intestinal damage by NSAIDs : enhanced intestinal permeability , inhibition of cyclooxygenase ( P36551 ) , enterohepatic recirculation , and formation of adducts . The effects of P35354 selective inhibitors , which appear to have better gastric tolerability when compared to nonselective NSAIDs , on normal and inflamed intestinal mucosa ( as in Crohn 's disease or ulcerative colitis ) are still largely unexplored . If P35354 inhibition plays a key role in suppressing the inflammatory process , recent evidence suggests that P35354 products are involved in maintaining the integrity of intestinal mucosa , in the healing of gastrointestinal ulcers and in the modulation of inflammatory bowel disease ( Q9UKU7 ) . Animal models of intestinal inflammation have so far yielded conflicting results on the effects of P35354 selective inhibitors on the intestinal mucosa . It is now clear that NSAIDs do not act through cyclooxygenase inhibition , but also have different targets such as nuclear factor-kappaB and/or peroxisome proliferator-activated receptors gamma . The peculiar pharmacological profile of each compound may help to explain the different impact of each NSAID on the inflammatory process and on Q9UKU7 . Notably , the salicylic acid derivative DB00244 is widely used in the treatment of Q9UKU7 and is believed to act through nuclear factor-kappaB inhibition . Although the use of P35354 selective inhibitors remains contraindicated in patients with Q9UKU7 , studying their effects on intestinal mucosa may offer new insights into their subcellulars mechanisms of action and open new avenues for the development of novel therapies for Q9UKU7 . 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 . 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 . P08195 -silencing impairs tumorigenicity of HeLa cells via modulation of galectin-3 and β-catenin signaling , and P08253 expression . P08195 is a type-II glycoprotein whose covalent-bound association with one of several described light chains yields a heterodimer mainly involved in large neutral amino acid transport . Likewise , it is well known that the heavy chain interacts with β-integrins mediating integrin-dependent events such as survival , proliferation , migration and even transformation . P08195 is a ubiquitous protein whose overexpression has been related to tumor development and progression . Stable silencing of P08195 in HeLa cells using an artificial miRNA impairs in vivo tumorigenicity and leads to an ineffective proliferation response to mitogens . P08195 colocalizes with β1-integrins and CD147 , but this interaction does not occur in lipid rafts in HeLa cells . Moreover , silenced cells present defects in integrin- ( Q05397 , Akt and P27361 /2 ) and hypoxia-dependent signaling , and reduced expression/activity of P08253 . These alterations seem to be dependent on the inappropriate formation of CD147/ P08195 /β1-integrin heterocomplexes on the cell surface , arising when CD147 can not interact with P08195 . Although extracellular galectin-3 accumulates due to the decrease in P08253 activity , galectin-3 signaling events are blocked due to an impaired interaction with P08195 , inducing an increased degradation of β-catenin . Furthermore , cell motility is compromised after protein silencing , suggesting that P08195 is related to tumor invasion by facilitating cell motility . Therefore , here we propose a molecular mechanism by which P08195 participates in tumor progression , favoring first steps of epithelial-mesenchymal transition by inhibition of β-catenin proteasomal degradation through Akt/GSK-3β signaling and enabling cell motility .

MH_train_1559

MH_train_1559

MH_train_1559

interacts_with DB01120?

multiple_choice

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 . High glucose suppresses expression of equilibrative nucleoside transporter 1 ( Q99808 ) in rat cardiac fibroblasts through a mechanism dependent on PKC-zeta and Q96HU1 kinases . Recently it was demonstrated that the elevated concentration of glucose but not lack of insulin is responsible for suppression of equilibrative nucleoside transporter ( Q99808 ) in diabetic rat cardiac fibroblasts ( CFs ) . The present study was undertaken to determine the signaling pathway utilized by glucose to regulate the expression of Q99808 in the primary culture of rat CFs . Pretreatment of CFs with Go 6983 , an isozyme non-selective PKC inhibitor , prevented the high glucose ( 25 mM ) effect on Q99808 mRNA level and nitrobenzylthioinosine ( NBTI ) -sensitive adenosine uptake . Similar effect was observed with a cell-permeable PKC-zeta pseudosubstrate , whereas Go 6976 a selective inhibitor of Ca(2+)-dependent P17252 and P05771 isozymes had little effect on high glucose-induced suppression of Q99808 mRNA level . Incubation of CFs with nitric oxide ( NO ) donors ( SNAPE , SNP ) or NO synthase inhibitors ( L-NAME , L-NMMA ) prior to exposition of CFs to high glucose did not change the glucose effect on Q99808 mRNA level . The high glucose-induced suppression of Q99808 expression was blocked by PD9859 ( an inhibitor of MEK ) , whereas neither wortmannin ( an inhibitor of PI3K ) nor rapamycin ( an inhibitor of P42345 ) affected the glucose action on Q99808 transcript level . Highly effective in preventing the high glucose effect on Q99808 mRNA level were GW 5074 ( an inhibitor of Raf kinase ) and SB 203580 ( selective p38 MAPK inhibitor ) . These findings indicate that high glucose suppresses the expression of Q99808 in CFs by NO independent manner involving the signaling through PKC-zeta , P04049 , MEK , and p38 MAPK pathways . Expression data analysis to identify biomarkers associated with asthma in children . Asthma is characterized by recurrent episodes of wheezing , shortness of breath , chest tightness , and coughing . It is usually caused by a combination of complex and incompletely understood environmental and genetic interactions . We obtained gene expression data with high-throughput screening and identified biomarkers of children 's asthma using bioinformatics tools . Next , we explained the pathogenesis of children 's asthma from the perspective of gene regulatory networks : DAVID was applied to perform Kyoto Encyclopedia of Genes and Genomes ( KEGG ) pathway enriching analysis for the top 3000 pairs of relationships in differentially regulatory network . Finally , we found that O96004 , Q16584 , P19838 , O60481 , P42226 , Q01094 , Q8IZL8 , Q15853 , and Q13951 may play important roles in children 's asthma initiation . On account of regulatory impact factor ( Q9HBH0 ) score , O96004 , Q13308 , and O60481 were the potential asthma-related factors . Our study provided some foundations of a strategy for biomarker discovery despite a poor understanding of the mechanisms underlying children 's asthma . 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 . 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 . Improvement of chloride transport defect by gonadotropin-releasing hormone ( DB00644 ) in cystic fibrosis epithelial cells . Cystic fibrosis ( CF ) , the most common autosomal recessive disease in Caucasians , is due to mutations in the P13569 gene . F508del , the most frequent mutation in patients , impairs P13569 protein folding and biosynthesis . The F508del- P13569 protein is retained in the endoplasmic reticulum ( ER ) and its traffic to the plasma membrane is altered . Nevertheless , if it reaches the cell surface , it exhibits a Cl(-) channel function despite a short half-life . Pharmacological treatments may target the F508del- P13569 defect directly by binding to the mutant protein or indirectly by altering cellular proteostasis , and promote its plasma membrane targeting and stability . We previously showed that annexine A5 ( AnxA5 ) directly binds to F508del- P13569 and , when overexpressed , promotes its membrane stability , leading to the restoration of some Cl(-) channel function in cells . Because Gonadotropin-Releasing Hormone ( DB00644 ) increases AnxA5 expression in some cells , we tested it in CF cells . We showed that human epithelial cells express DB00644 -receptors ( P30968 ) and that DB00644 induces an AnxA5 overexpression and an increased Cl(-) channel function in F508del- P13569 cells , due to an increased stability of the protein in the membranes . Beside the numerous physiological implications of the P30968 expression in epithelial cells , we propose that a topical use of DB00644 is a potential treatment in CF . 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 . 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 . Decreased mitochondrial carnitine translocase in skeletal muscles impairs utilization of fatty acids in insulin-resistant patients . P01308 resistance ( IR ) and its health consequences ( diabetes , hypertension , cardiovascular disease , obesity etc. ) affect between 25 and 35 % of Westernized populations . Decreased fatty acid ( FA ) oxidation in skeletal muscle is implicated in obesity-related IR . DB00583 -acylcarnitine translocase ( O43772 ) transports long-chain FAs both into mitochondria ( as carnitine esters for energy-generating processes ) and out of mitochondria . To determine whether O43772 activity correlates with decreased FA oxidation we measured O43772 concentrations in cellular and mitochondrial extracts from the skeletal muscle of 19 obese IR individuals and of 19 lean controls . We also evaluated carnitine transport in skeletal muscle mitochondria in both groups . Mitochondrial O43772 was decreased at translational and transductional level , and carnitine-carnitine and acylcarnitine-carnitine exchange rates were significantly lower in IR subjects . Aberrant acylcarnitine flux into mitochondria was not correlated with decreased activity of other components of the mitochondrial carnitine system ( i.e. , carnitine palmitoyl transferase-I and II ) . Our data suggest that by restraining entry of FA-coenzyme A into mitochondria , low O43772 levels increase cytosolic FA levels and their incorporation into glycerolipids . The low level of O43772 in IR muscle may contribute to the elevated muscle concentrations of triglycerides , diacylglycerol , and FA-coenzyme A characteristic of IR muscle . Effects of maternal smoking on the placental expression of genes related to angiogenesis and apoptosis during the first trimester . OBJECTIVE : Maternal cigarette smoking is reportedly associated with miscarriage , fetal growth restriction and placental abruption , and is paradoxically associated with a decreased risk of developing preeclampsia . In the present study , we investigated the gene expression levels of villous tissues in early gestation . We compared the expression levels of the genes related to angiogenesis and apoptosis in the villous tissues obtained from smoking and non-smoking pregnant women . MATERIALS AND METHODS : We collected villous tissue samples from 57 women requesting surgical termination due to non-medical reasons at 6-8 weeks of gestation . The maternal cigarette smoking status was evaluated by the level of serum cotinine and patients were divided into active smokers and non-smokers by the serum cotinine level . The placental levels of P15692 , P49763 , P17948 , Q16665 , P04637 , Q07812 and P10415 mRNA were quantified by real time PCR . RESULTS : The gene expression level of P49763 and Q16665 in the active smoker group was significantly higher than that in the non-smoker group . We did not observe any significant differences in the P15692 or P17948 expression between the groups . In active smoker group , the gene expression levels of P04637 and Q07812 were significantly higher than those in the non-smoker group . The ratio of Q07812 / P10415 mRNA in the active smoker group was significantly higher than that in the non-smoker group . CONCLUSIONS : Our findings revealed that smoking might affect the placenta during early pregnancy . Maternal cigarette smoking in early pregnancy may be associated with villus hypoxia , which may influence angiogenesis and apoptosis . Differential neurotrophic regulation of sodium and calcium channels in an adult sympathetic neuron . Adult neuronal phenotype is maintained , at least in part , by the sensitivity of individual neurons to a specific selection of neurotrophic factors and the availability of such factors in the neurons ' environment . Nerve growth factor ( P01138 ) increases the functional expression of Na(+) channel currents ( I(Na) ) and both N- and L-type Ca(2+) currents ( I(Ca,N) and I(Ca,L) ) in adult bullfrog sympathetic ganglion ( BFSG ) B-neurons . The effects of P01138 on I(Ca) involve the mitogen-activated protein kinase ( MAPK ) pathway . Prolonged exposure to the ganglionic neurotransmitter luteinizing hormone releasing hormone ( P01148 ) also increases I(Ca,N) but the transduction mechanism remains to be elucidated as does the transduction mechanism for P01138 regulation of Na(+) channels . We therefore exposed cultured BFSG B-neurons to chicken II P01148 ( 0.45 microM ; 6-9 days ) or to P01138 ( 200 ng/ml ; 9-10 days ) and used whole cell recording , immunoblot analysis , and ras or rap-1 pulldown assays to study effects of various inhibitors and activators of transduction pathways . We found that 1 ) P01148 signals via ras-MAPK to increase I(Ca,N) , 2 ) this effect is mediated via protein kinase C-beta ( P05771 -IotaIota ) , 3 ) protein kinase A ( PKA ) is necessary but not sufficient to effect transduction , 4 ) P01138 signals via phosphatidylinositol 3-kinase ( PI3K ) to increase I(Na) , and 5 ) long-term exposure to P01148 fails to affect I(Na) . Thus downstream signaling from P01148 has access to the ras-MAPK pathway but not to the PI3K pathway . This allows for differential retrograde and anterograde neurotrophic regulation of sodium and calcium channels in an adult sympathetic neuron . 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 . DB08439 sodium , an injectable P35354 -specific inhibitor , does not affect unfractionated heparin-regulated blood coagulation parameters . The objective of this study was to evaluate the potential for hemostatic interaction between a full analgesic dose of parecoxib sodium ( parecoxib ) , a prodrug of the P35354 specific inhibitor valdecoxib , and unfractionated heparin ( UFH ) in healthy male subjects . This open-label , single-center study comprised two treatment periods . In treatment period I , fasted , eligible subjects ( n = 18 ) received a UFH bolus ( 4000 U ) followed by a 36-hour UFH infusion ( start dose 10-14 U/kg ) . Activated partial thromboplastin time ( aPTT ) , prothrombin time ( PT ) , and platelet counts were measured at regular intervals up to 24 hours after the end of the UFH infusion . After a 2-day washout , patients randomized to treatment period II received a full analgesic dosage of parecoxib 40 mg bid intravenously ( IV ) for 6 days ( n = 18 ) , with concomitant UFH ( same regimen as treatment period I ) on day 5 ( n = 18 ) . APTT , PT , and platelet counts were evaluated at regular intervals up to 24 hours after UFH infusion . Coadministration of parecoxib 40 mg bid IV with UFH ( treatment period II ) had no significant effect on aPTT , PT , or platelet counts , which were similar to those of participants receiving UFH alone ( treatment period I ) at all time points . These results show that a full analgesic dose of parecoxib , a P35354 -specific inhibitor available for parenteral administration , can be coadministered with UFH without affecting blood coagulation parameters . Therefore , parecoxib may be administered to patients who are receiving UFH for thromboprophylaxis . 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 . Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 -induced survival . This inhibition can not be overcome by increasing concentrations of P05113 and is not due to the blocking of Na+ channels by lidocaine . Here we report that one class of K+ channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 . In contrast , increasing concentrations of P08700 or granulocyte-macrophage P04141 overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 -sensitive K+ channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [3H]glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases . Gene expression profiles of B-lineage adult acute lymphocytic leukemia reveal genetic patterns that identify lineage derivation and distinct mechanisms of transformation . PURPOSE : To characterize gene expression signatures in acute lymphocytic leukemia ( ALL ) cells associated with known genotypic abnormalities in adult patients . EXPERIMENTAL DESIGN : Gene expression profiles from 128 adult patients with newly diagnosed ALL were characterized using high-density oligonucleotide microarrays . All patients were enrolled in the Italian GIMEMA multicenter clinical trial 0496 and samples had > 90 % leukemic cells . Uniform phenotypic , cytogenetic , and molecular data were also available for all cases . RESULTS : T-lineage ALL was characterized by a homogeneous gene expression pattern , whereas several subgroups of B-lineage ALL were evident . Within B-lineage ALL , distinct signatures were associated with Q03164 / P51825 and P15923 / P40424 gene rearrangements . Expression profiles associated with Q03164 / P51825 and P15923 / P40424 are similar in adults and children . P11274 / P00519 + gene expression pattern was more heterogeneous and was most similar to ALL without known molecular rearrangements . We also identified a set of 83 genes that were highly expressed in leukemia blasts from patients without known molecular abnormalities who subsequently relapsed following therapy . Supervised analysis of kinase genes revealed a high-level P36888 expression in a subset of cases without molecular rearrangements . Two other kinases ( P05771 and Q08345 ) were highly expressed in cases without molecular rearrangements , as well as in P11274 / P00519 -positive ALL . CONCLUSIONS : Genomic signatures are associated with phenotypically and molecularly well defined subgroups of adult ALL . Genomic profiling also identifies genes associated with poor outcome in cases without molecular aberrations and specific genes that may be new therapeutic targets in adult ALL . L-carnitine preserves endothelial function in a lamb model of increased pulmonary blood flow . BACKGROUND : In our model of a congenital heart defect ( Q8NE62 ) with increased pulmonary blood flow ( PBF ; shunt ) , we have recently shown a disruption in carnitine homeostasis , associated with mitochondrial dysfunction and decreased endothelial nitric oxide synthase ( P29474 ) /heat shock protein (Hsp)90 interactions that contribute to P29474 uncoupling , increased superoxide levels , and decreased bioavailable nitric oxide ( NO ) . Therefore , we undertook this study to test the hypothesis that L-carnitine therapy would maintain mitochondrial function and NO signaling . METHODS : Thirteen fetal lambs underwent in utero placement of an aortopulmonary graft . Immediately after delivery , lambs received daily treatment with oral L-carnitine or its vehicle . RESULTS : L- DB00583 -treated lambs had decreased levels of acylcarnitine and a reduced acylcarnitine:free carnitine ratio as compared with vehicle-treated shunt lambs . These changes correlated with increased carnitine acetyl transferase ( P43155 ) protein and enzyme activity and decreased levels of nitrated P43155 . The lactate:pyruvate ratio was also decreased in L-carnitine-treated lambs . Hsp70 protein levels were significantly decreased , and this correlated with increases in P29474 /Hsp90 interactions , NOS activity , and NOx levels , and a significant decrease in P29474 -derived superoxide . Furthermore , acetylcholine significantly decreased left pulmonary vascular resistance only in L-carnitine-treated lambs . CONCLUSION : L- DB00583 therapy may improve the endothelial dysfunction noted in children with CHDs and has important clinical implications that warrant further investigation . Functions of flt3 in zebrafish hematopoiesis and its relevance to human acute myeloid leukemia . P07333 -like tyrosine kinase 3 ( P36888 ) is expressed in human hematopoietic stem and progenitor cells ( HSPCs ) but its role during embryogenesis is unclear . In acute myeloid leukemia ( AML ) , internal tandem duplication ( ITD ) of P36888 at the juxtamembrane ( JMD ) and tyrosine kinase ( TKD ) domains ( P36888 -ITD(+) ) occurs in 30 % of patients and is associated with inferior clinical prognosis . TKD mutations ( P36888 -TKD(+) ) occur in 5 % of cases . We made use of zebrafish to examine the role of flt3 in developmental hematopoiesis and model human P36888 -ITD(+) and P36888 -TKD(+) AML . Zebrafish flt3 JMD and TKD were remarkably similar to their mammalian orthologs . Morpholino knockdown significantly reduced the expression of l-plastin ( pan-leukocyte ) , csf1r , and mpeg1 ( macrophage ) as well as that of c-myb ( definitive HSPCs ) , lck , and rag1 ( T-lymphocyte ) . Expressing human P36888 -ITD in zebrafish embryos resulted in expansion and clustering of myeloid cells ( pu.1(+) , mpo(+) , and cebpα(+) ) which were ameliorated by DB05213 and associated with stat5 , erk1/2 , and akt phosphorylation . Human P36888 -TKD ( D835Y ) induced significant , albeit modest , myeloid expansion resistant to DB05213 . This study provides novel insight into the role of flt3 during hematopoiesis and establishes a zebrafish model of P36888 -ITD(+) and P36888 -TKD(+) AML that may facilitate high-throughput screening of novel and personalized agents . Eosinophilic oesophagitis : a common cause of dysphagia in young adults ? Eosinophilic oesophagitis ( EO ) is an increasingly recognised chronic , relapsing inflammatory condition of the oesophagus . There has been a mini-epidemic of EO in the last decade . The incidence of this condition is higher in children and is commoner in males . There is either a family or personal history of atopic conditions present in a significant number of patients and can also be familial in up to 10 % . The classical symptom in an adult is chronic , intermittent solid-food dysphagia and food impaction , often necessitating emergency endoscopic removal . Despite the history of dysphagia for a number of years , patients remain well with no weight loss , which can mislead clinicians to diagnose a functional problem with a resulting delay in the diagnosis . There are various endoscopic features of EO ; commonly multiple rings and linear furrows , though these can be subtle and the mucosa may be macroscopically normal . The hallmark of this condition is the histological presence of > or =15 eosinophils/high power field ( HPF ) in the oesophageal mucosa . Therapeutic options include avoidance of dietary allergens , topical or systemic steroids , Montelukast , DB06612 ( anti- P05113 antibody ) and endoscopic dilation of strictures unresponsive to medical therapy . Bcl-2/Bcl-xL inhibition increases the efficacy of MEK inhibition alone and in combination with P19957 kinase inhibition in lung and pancreatic tumor models . Although mitogen-activated protein ( Q96HU1 ) -extracellular signal-regulated kinase ( P29323 ) kinase ( MEK ) inhibition is predicted to cause cell death by stabilization of the proapoptotic Q7L3V2 O43521 , the induction of apoptosis is often modest . To determine if addition of a Bcl-2 family inhibitor could increase the efficacy of a MEK inhibitor , we evaluated a panel of 53 non-small cell lung cancer and pancreatic cancer cell lines with the combination of navitoclax ( DB05764 ) , a Bcl-2/Bcl-xL ( P10415 / Q07817 ) antagonist , and a novel Q96HU1 kinase ( MEK ) inhibitor , G-963 . The combination is synergistic in the majority of lines , with an enrichment of cell lines harboring P01116 mutations in the high synergy group . Cells exposed to G-963 arrest in P55008 and a small fraction undergo apoptosis . The addition of navitoclax to G-963 does not alter the kinetics of cell-cycle arrest , but greatly increases the percentage of cells that undergo apoptosis . The G-963/navitoclax combination was more effective than either single agent in the P01116 mutant H2122 xenograft model ; O43521 stabilization and PARP cleavage were observed in tumors , consistent with the mechanism of action observed in cell culture . Addition of the phosphatidylinositol 3-kinase ( PI3K , P42336 ) inhibitor P16260 -0941 to this treatment combination increases cell killing compared with double- or single-agent treatment . Taken together , these data suggest the efficacy of agents that target the MAPK and PI3K pathways can be improved by combination with a Bcl-2 family inhibitor . P36888 inhibition as therapy in acute myeloid leukemia : a record of trials and tribulations . Acute myeloid leukemia ( AML ) is a hematologic malignancy with a poor prognosis . Approximately one quarter of the patients with AML also carry an internal tandem duplication ( ITD ) mutation in the gene encoding P07333 -like tyrosine kinase 3 ( P36888 ) , which has a significantly deleterious impact on prognosis . The ITD mutation renders P36888 constitutively active and leads to uncontrolled proliferation of the leukemic blast . Over the course of the last decade , a variety of compounds have been developed in preclinical and clinical studies as potent inhibitors of P36888 . Many of the earlier agents under investigation , such as lestaurtinib , midostaurin , and sunitinib , were initially developed as inhibitors of other tyrosine kinases and as targeted therapies in a variety of malignancies . These compounds have been demonstrated to have some efficacy in clinical trials of AML , mainly manifesting as transient decreases in circulating blasts correlating with effective in vivo suppression of the P36888 target . Nevertheless , the cumbersome pharmacokinetics of some compounds and the suboptimal specificity and potency of others have limited their therapeutic efficacy . In the last few years , newer , more potent and specific agents have been under investigation , with the leading example being DB05213 . This agent has shown significant promise in early phases of clinical investigation , and is currently in more advanced clinical trials . Hope remains that P36888 inhibition will be become an effective therapeutic adjunct to our current treatment approach to AML . 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 . Periadventitial adipose tissue impairs coronary endothelial function via P05771 -dependent phosphorylation of nitric oxide synthase . Endogenous periadventitial adipose-derived factors have been shown to contribute to coronary vascular regulation by impairing endothelial function through a direct inhibition of endothelial nitric oxide synthase ( P29474 ) . However , our understanding of the underlying mechanisms remains uncertain . Accordingly , this study was designed to test the hypothesis that periadventitial adipose tissue releases agents that attenuate coronary endothelial nitric oxide production via a protein kinase C ( PKC ) -beta-dependent mechanism . Isometric tension studies were conducted on isolated canine circumflex coronary arteries with and without natural amounts of periadventitial adipose tissue . Adipose tissue significantly diminished coronary endothelial-dependent vasodilation and nitric oxide production in response to bradykinin and acetylcholine . The selective inhibition of endothelial P05771 with ruboxistaurin ( 1 microM ) abolished the adipose-induced impairment of bradykinin-mediated coronary vasodilation and the endothelial production of nitric oxide . Western blot analysis revealed a significant increase in P29474 phosphorylation at the inhibitory residue DB00156 (495) in arteries exposed to periadventitial adipose tissue . This site-specific phosphorylation of P29474 was prevented by the inhibition of P05771 . These data demonstrate that periadventitial adipose-derived factors impair coronary endothelial nitric oxide production via a P05771 -dependent , site-specific phosphorylation of P29474 at DB00156 (495) . 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 . 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 . 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 . 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 . 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 . Ameliorative effect of combined administration of inducible nitric oxide synthase inhibitor with cyclooxygenase-2 inhibitors in neuropathic pain in rats . OBJECTIVES : The objective of this study was to examine the effects of rofecoxib , meloxicam , both cyclooxygenase-2 ( P35354 ) inhibitors and aminoguanidine hydrochloride , an inducible nitric oxide synthase ( P35228 ) inhibitor and their combinations in neuropathic pain in rats . METHODS : Neuropathy was induced by chronic constriction injury ( CCI ) of right sciatic nerve under ketamine anesthesia in rats . Effect of ED(50) of aminoguanidine hydrochloride , rofecoxib and meloxicam administered orally was investigated using behavioral tests . Effect of combinations of aminoguanidine hydrochloride with rofecoxib and meloxicam was also investigated in neuropathic pain employing behavioral tests . RESULTS : Behavioral tests , mechanical , thermal and cold stimuli confirmed the development of neuropathic pain after CCI . DB02533 hydrochloride , rofecoxib and meloxicam when administered alone , produced significant increase in paw withdrawal threshold to mechanical stimuli at 6 h in ipsilateral hind paw after CCI . Co-administration of aminoguanidine hydrochloride ( 30 mg/kg ) with rofecoxib ( 1.31 mg/kg ) and meloxicam ( 1.34 mg/kg ) was also found to produce significant increase in paw withdrawal latencies to mechanical stimuli at 6 h . Combined administration of aminoguanidine hydrochloride with meloxicam and rofecoxib produced significant rise in pain threshold for mechanical hyperalgesia in ipsilateral hind paw when compared with the groups treated with aminoguanidine hydrochloride , meloxicam and rofecoxib alone . CONCLUSION : Co-administration of meloxicam and rofecoxib with aminoguanidine hydrochloride may be an alternative approach for the treatment of neuropathic pain . Permanent neonatal diabetes mellitus in China . BACKGROUND : Permanent neonatal diabetes mellitus ( PNDM ) is a rare disease , which is defined as the onset of diabetes before the age of 6 months with persistence through life . Infants with Q14654 or Q09428 genetic mutations may respond to oral sulfonylurea therapy . Currently , there are limited studies about the genetic analysis and long-term follow-up of PNDM . CASE PRESENTATION : We report four cases of PNDM . None of the infants or their parents had P01308 , Q14654 , or Q09428 genetic mutations . One infant underwent continuous subcutaneous insulin infusion ( CSII ) and the other infants underwent multiple injections of insulin ( MII ) . In these infants , PNDM persisted from 35 months to 60 months of follow-up . Three infants maintained fairly stable blood sugar levels , and one infant had poor sugar control . CONCLUSIONS : We suggest that all of the infants with PNDM should undergo genetic evaluation . For infants without Q14654 and Q09428 genetic mutations , oral sulfonylurea should not be considered as treatment . CSII is a useful method for overcoming the difficulties of diabetes , and it may also improve the quality of life of both infants and their parents . Use of P01148 antagonists in reproductive medicine . Gonadotrophin-releasing hormone ( DB00644 ) plays a key role in the secretion of gonadotrophins , follicle-stimulating hormone ( DB00094 ) and luteinizing hormone ( LH ) , which regulate steroidogenesis and folliculogenesis . Two DB00644 antagonists , DB00050 and DB06785 , deprived of histaminergic side-effects , have been introduced into ovarian stimulation protocols to prevent premature LH surges and proved their safety in clinical trials . At present , most of the published studies have not found significant differences in follicular recruitment , oocyte quality , and so on , except for a decrease in pregnancy and implantation rates in in vitro fertilization and embryo transfer ( IVF-ET ) cycles when the DB00644 antagonist rather than the agonist was used . This decrease in pregnancy rates was in relation with a necessary learning curve of the physicians . Another possibility is the impact of the DB00644 antagonist on endometrium through its P30968 ; this effect was cancelled after cryopreserved embryo transfers because the pregnancy rates were similar between DB00644 antagonist and agonist in this case . DB00644 antagonists were also interesting in poor responders and polycystic ovarian syndrome , where the agonists have not permitted to obtain the better results in IVF-ET cycles . Similarly , the DB00644 antagonists could prevent the LH surge in the intrauterine insemination cycles . P35354 inhibition attenuates endotoxin-induced downregulation of organic anion transporters in the rat renal cortex . Renal excretion of organic anions such as DB00345 is reduced during severe sepsis and following ischemia/reperfusion injury . In order to better define the pathophysiology of sepsis-associated renal tubular dysfunction we measured the effect of lipopolysaccharide on renocortical organic anion transporter ( P04181 ) expression in the rat . DB00917 ( DB00917 ) downregulates OATs in vitro , therefore , we also evaluated the effect of the cyclooxygenase ( P36551 ) -2 inhibitor parecoxib on this process . Endotoxemia caused a time- and dose-dependent decrease of Q4U2R8 and Q8TCC7 expression that paralleled increased renocortical P35354 expression and DB00917 formation . Pretreatment with parecoxib decreased endotoxin-stimulated PGE(2) formation . DB08439 attenuated Q4U2R8 and Q8TCC7 gene repression in the rat kidney following endotoxin treatment and during ischemia/reperfusion-induced acute renal injury . P35354 inhibition improved the creatinine clearance in lipopolysaccharide-treated rats but not after ischemia/reperfusion-induced acute renal injury . The decreased clearance of DB00345 in rats following endotoxin- or ischemia/reperfusion-induced renal injury was improved by parecoxib . Our findings show that P35354 derived prostanoids downregulate OATs during lipopolysaccharide-induced acute renal injury . Combination of DB05210 and gefitinib induces apoptosis of triple-negative breast cancer cells through the PI3K/AKT- P42345 pathway . To investigate the apoptotic mechanism of triple-negative breast cancer ( TNBC ) cells induced by gefitinib and PI3K inhibitor DB05210 . MDA-MB-231 , MDA-MB-436 , and MCF-7 cells were incubated with 0.1 μmol/l gefitinib , 1 μmol/l gefitinib , 10 μmol/l gefitinib , 1 μmol/l DB05210 , 0.1 μmol/l gefitinib+1 μmol/l DB05210 , 1 μmol/l gefitinib+1 μmol/l DB05210 , and 10 μmol/l gefitinib+1 μmol/l DB05210 . Then , cell viability and survival were determined using an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide ( MTT ) assay and Hoechst staining . The apoptosis-related factors and phosphoinositide-3-kinase/protein kinase B , the mammalian target of rapamycin ( PI3K/AKT- P42345 ) signaling pathway-related factors were detected by western blot . For TNBC cells , cell viability or survival was not significantly inhibited by gefitinib or DB05210 alone ; however , marked cell apoptosis was noted in the gefitinib and DB05210 combination groups , and this effect was dose dependent . Also , the expressions of apoptosis markers , such as cleaved caspase-3 , Bcl-2/Bax , were altered by the gefitinib and DB05210 combination . Moreover , phosphorylated AKT ( p-AKT ) and 70 kDa ribosomal protein S6-kinase ( p-p70S6K ) were also inhibited by the gefitinib and DB05210 combination , which may be responsible for the apoptosis . Gefitinib combined with DB05210 could induce cell apoptosis in TNBC cells and this effect was mediated through the P00533 -PI3K-AKT- P42345 -p70S6K pathway . Our studies have set the stage for future clinical trials of TNBC therapy by the combination of gefitinib and DB05210 . [ Eosinophilic esophagitis -- pathogenesis , clinical presentation and therapeutic management ] . Eosinophilic esophagitis ( EE ) is a relatively new , chronic , TH 2-type allergic inflammation of the esophagus . EE occurs more frequently in men . Allergic diseases such as asthma or atopic dermatitis are present in 50-70 % of patients or their relatives . In adults , the most common presenting symptom of EE is dysphagia , with or without food bolus impaction . Endoscopic findings of EE include mucosal furrows , corrugated or concentric rings or ridges in the esophagus ( " feline esophagus " ) , with or without tiny whitish exudates . The diagnosis is confirmed by the observation of high counts of eosinophils in the esophageal epithelium ( at least 24 /HPF ) . The cornerstones of medical therapy are either topical or systemic corticosteroids . Additional therapies included leukotriene receptor antagonists ( montelukast ) and P05113 blockers ( DB06612 ) . Complications of EE such as esophageal strictures should be carefully dilated using either bougies or a balloon . Currently it is still not known whether the late complications of EE can be prevented by the use of anti-inflammatory agents and this can only be demonstrated through further long-term follow-up studies . [ Effect of inducible nitric oxide synthase on pancreas islet apoptosis in rats ] . AIM : To investigate the effect and mechanism of cytokine and inducible nitric oxide synthase on apoptosis and function of rat pancreas islets cultured in vitro . METHODS : Islets from Wistar rats were cultured in vitro and divided randomly into four groups : blank control group , cytokine group of islets cultured with P01375 +IL-1beta , aminoguanidine ( AG ) group of islets cultured with aminoguanidine , and AG + cytokine group of islets cultured with P01375 +IL-1beta and aminoguanidine . The nutrient fluid nitric oxide level and islets P29474 / P35228 activity were detected by test kit and the expressions of P35228 mRNA and apoptosis related gene ( Bax , Bcl-2 ) were evaluated by RT-PCR . The viability of the islets was examined by AO/EB staining and the function of the islets was detected by insulin secretion index assay . RESULTS : After co-cultured with cytokines IL-1beta and P01375 , the expression and activity of P35228 in islet tissues enhanced ( 38.93+/-4.72 ) U/mL and the concentration of NO in medium increased remarkably(313.0+/-35.4) mol/L.The survival rate of cells and the insulin secretion index decreased with the up-regulation of proapoptosis gene and down-regulation of anti-apoptosis gene . But the activity of P29474 remained unchanged . DB02533 reduced the cell apoptosis and increased the survival rate and insulin secretion index , and the activity of P35228 was inhibited . CONCLUSION : P35228 plays an important role in the apoptosis of islets cultured by cytokines P01375 and IL1-beta . DB02533 prevents the islets from the damage of P35228 , alleviates the impairment of cytokines to islets , lessens the cell apoptosis and ameliorates the survival and function of islets .

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interacts_with DB01418?

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Glioblastoma with ovarian teratoma having N-methyl-D-aspartate receptor ( NMDAR ) antibody in P04141 -- a case report . A 54-year-old woman presented with complex partial seizure with impaired consciousness . Brain Q9BWK5 revealed a high intensity lesion on P24752 -weighted and FLAIR images in the left temporal lobe , indicating limbic encephalitis . CT and Q9BWK5 of the pelvis showed right ovarian teratoma . The cerebrospinal fluid ( P04141 ) were positive for antibodies against the GluRε2 , GluRδ2 , and antibodies against Q9UHB4 + Q13224 heteromers . On the basis of these data , anti-N-methyl-D-aspartate receptor ( NMDAR ) encephalitis associated with ovarian teratoma was suspected , and the right ovariectomy was performed . Six months after onset , brain biopsy from the right temporal lobe led to a diagnosed of glioblastoma . This is the first glioblastoma case with ovarian teratoma having autoantibodies against GluR and Q9UHB4 + Q13224 heteromers in P04141 . We suggest that patients with NMDAR antibodies should be carefully diagnosed with anti-NMDAR encephalitis . Genetic markers associated with abstinence length in alcohol-dependent subjects treated with acamprosate . DB00659 supports abstinence in some alcohol-dependent subjects , yet predictors of response are unknown . To identify response biomarkers , we investigated associations of abstinence length with polymorphisms in candidate genes in glycine and glutamate neurotransmission pathways and genes previously implicated in acamprosate response . Association analyses were conducted in the discovery sample of 225 alcohol-dependent subjects treated with acamprosate for 3 months in community-based treatment programs in the United States . Data from 110 alcohol-dependent males treated with acamprosate in the study PREDICT were used for replication of the top association findings . Statistical models were adjusted for relevant covariates , including recruitment site and baseline clinical variables associated with response . In the discovery sample , shorter abstinence was associated with increased intensity of alcohol craving and lower number of days between the last drink and initiation of acamprosate treatment . After adjustment for covariates , length of abstinence was associated with the Q13224 rs2058878 ( P=4.6 × 10(-5) ) . In the replication sample , shorter abstinence was associated with increased craving , increased depressive mood score and higher alcohol consumption . Association of abstinence length with Q13224 rs2058878 was marginally significant ( P=0.0675 ) ; as in the discovery sample , the minor A allele was associated with longer abstinence . Furthermore , rs2300272 , which is in strong linkage disequilibrium with rs2058878 , was also associated with abstinence length ( P=0.049 ) . This is the first report of a replicated association of genetic markers with the length of abstinence in acamprosate-treated alcoholics . Investigation of the underlying mechanisms of this association and its usefulness for individualized treatment selection should follow . Granulocyte-colony stimulating factor attenuates oligomeric amyloid β neurotoxicity by activation of neprilysin . Soluble oligomeric amyloid β ( oAβ ) causes synaptic dysfunction and neuronal cell death , which are involved in the pathogenesis of Alzheimer 's disease ( AD ) . The hematopoietic growth factor granulocyte-colony stimulating factor ( DB00099 ) is expressed in the central nervous system ( CNS ) and drives neurogenesis . Here we show that G- P04141 attenuated oAβ neurotoxicity through the enhancement of the enzymatic activity of Aβ-degrading enzyme neprilysin ( NEP ) in neurons , while the NEP inhibitor thiorphan abolished the neuroprotection . Inhibition of Q13163 / Q13164 , a major downstream effector of G- P04141 signaling , also ablated neuroprotective effect of DB00099 . Furthermore , intracerebroventricular administration of G- P04141 enhanced NEP enzymatic activity and clearance of Aβ in P05067 / P49768 transgenic mice . Thus , we propose that G- P04141 may be a possible therapeutic strategy against AD . [ 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 . 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 . 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 . 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 . [ Detection of alterations of dihydrofolate reductase gene in folate-resistant leukemia cells by in vitro enzymatic amplification ] . Three methods for analyzing the products of polymerase chain reaction were applied to detect complex alterations of dihydrofolate reductase ( P00374 ) gene , in order to assess their value in detection of folate-resistance in leukemia cells . A single point mutation in the second position of codon 31 , a T-to-C transition , in trimetrexate ( DB01157 ) resistant Q14C87 -3/TMQ200 cells was detected by either allele-specific oligonucleotide hybridization or restriction pattern of the PCR product . These two analyses allowed us to detect not only the presence of the mutation , but also the amplification of the mutated gene in DB01157 -methotrexate ( MTX ) doubly-resistant Q14C87 -3/TMQ200-MTX500 cells . The base change was confirmed by direct sequencing method of the PCR product . Using these analyses of the PCR product , the complex alterations of P00374 gene are to be examined in leukemic patient cells . Expression of dihydrofolate reductase and multidrug resistance genes in trimetrexate-resistant human leukemia cell lines . Exposure of Q14C87 -3 human leukemic cells in culture to a lipophilic antifolate , trimetrexate ( DB01157 ) , resulted in the development of sublines resistant to antifolates as well as to drugs related to multidrug resistance . The DB01157 -resistant sublines had an increase in dihydrofolate reductase ( P00374 ) activity and overexpression of P-glycoprotein . In these sublines , neither the P00374 gene nor the P08183 gene were amplified . In these cells , P00374 transcripts were also not overexpressed but P00374 protein was increased , indicative of translational or post-translational control of P00374 activity . In contrast , P08183 transcripts were found to be overexpressed , in parallel with P-glycoprotein production . Therefore , increases in P-glycoprotein appear controlled at the transcriptional level . These data support evidence that DB01157 produced two phenotypic changes independently : the former probably from folate deficiency and the latter from the lipophilic nature of the compound . 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 . 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 . 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 . 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 . 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 . GM- P04141 priming drives bone marrow-derived macrophages to a pro-inflammatory pattern and downmodulates DB00917 in response to O60603 ligands . In response to pathogen recognition by Toll-like receptors ( TLRs ) on their cell surface , macrophages release lipid mediators and cytokines that are widely distributed throughout the body and play essential roles in host responses . Granulocyte macrophage colony-stimulating factor ( GM- P04141 ) is important for the immune response during infections to improve the clearance of microorganisms . In this study , we examined the release of mediators in response to O60603 ligands by bone marrow-derived macrophages ( BMDMs ) primed with GM- P04141 . We demonstrated that when stimulated with O60603 ligands , non-primed BMDMs preferentially produced PGE(2) in greater amounts than Q06643 (4) . However , GM- P04141 priming shifted the release of lipid mediators by BMDMs , resulting in a significant decrease of PGE(2) production in response to the same stimuli . The decrease of PGE(2) production from primed BMDMs was accompanied by a decrease in PGE-synthase mRNA expression and an increase in P01375 -α and nitric oxide ( NO ) production . Moreover , some GM- P04141 effects were potentiated by the addition of IFN-γ . Using a variety of O60603 ligands , we established that PGE(2) release by GM- P04141 -primed BMDMs was dependent on O60603 co-receptors ( Q15399 , Q9Y2C9 ) , P08571 , MyD88 and the nuclear translocation of NFκB but was not dependent on peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) activation . Indeed , GM- P04141 priming enhanced O60603 , O00206 and MyD88 mRNA expression and phospho-IκBα formation . These findings demonstrate that GM- P04141 drives BMDMs to present a profile relevant to the host during infections . DB04892 . DB04892 , a derivative of physostigmine , was first described as an inhibitor of acetylcholinesterase ( P22303 ) and was shown to improve cognition in various experimental paradigms in rodents and dogs . It was clinically tested for Alzheimer 's disease , with moderate success in initial Phase II studies . DB04892 deserves attention for an additional quality of action : in addition to inhibiting P22303 , it modulates the amount of beta-amyloid precursor protein ( P05067 ) in neuronal cell culture by reducing P05067 translation . This effect probably involves interaction of phenserine with a regulatory element in the 5'-untranslated region of the P05067 gene that controls P05067 expression . DB04892 apparently reduces translational efficiency of P05067 mRNA into protein , a process that may involve an interaction with iron and/or an iron-responsive element . As a consequence , phenserine reduces beta-amyloid peptide ( Abeta ) formation in vitro and in vivo . DB04892 is also unique because of differing actions of its enantiomers : DB04892 is the active enantiomer for inhibition of P22303 , whereas (+)-phenserine ( ' posiphen ' ) has weak activity as an P22303 inhibitor and can be dosed much higher . Both enantiomers are equipotent in downregulating P05067 expression . (+)-Posiphen may be a promising drug , either alone or in combination with DB04892 , to attenuate the progression of Alzheimer 's disease . DB01373 as a mediator between erythropoietin and protein tyrosine phosphatase 1B . Erythropoietin ( Epo ) is crucial for promoting the survival , proliferation , and differentiation of mammalian erythroid progenitors . The central role played by tyrosine phosphorylation of erythropoietin receptor ( EpoR ) in Epo-cell activation has focused attention on protein tyrosine phosphatases ( PTPs ) as candidates implicated in the pathogenesis of the resistance to therapy with human recombinant Epo . Prototypic member of the PTP family is P18031 , which has been implicated in the regulation of EpoR signaling pathways . In previous reports we have shown that P18031 is reciprocally modulated by Epo in undifferentiated UT-7 cell line . However , no information is available with respect to the modulation of this phosphatase in non-Epo depending cells or at late stages of erythroid differentiation . In order to investigate these issues we induced UT-7 cells to differentiate and studied their P18031 expression pattern . Simultaneous observations were performed in TF-1 cells which can be cultured either with GM- P04141 , P08700 or Epo . We found that Epo induced P18031 cleaveage in TF-1 and differentiated UT-7 cells . This pattern of P18031 modulation may be due to an increased Q13507 / Q9Y210 expression ratio which could explain the larger and sustained calcium response to Epo and calpain activation in Epo treated TF-1 and differentiated UT-7 cells . Thyroid-stimulating antibodies in sera from patients with Graves ' disease are heterogeneous in epitope recognition . Two synthetic peptides , P354-14 ( amino acid nos. 354 to 367 ) and P338-16 ( nos. 338 to 353 ) , corresponding to the partial amino acid sequences of the hTSH receptor structure were studied for their ability to bind specifically serum IgGs from patients with Graves ' disease and to inhibit thyroid stimulating DB00024 receptor antibody ( P16473 SAb ) activity . IgG binding was measured by an ELISA using sera from 102 Graves ' , 20 Hashimoto patients , and 9 normal subjects . Both peptides showed significantly increased IgG binding of Graves ' sera compared with those of Hashimoto and normal sera . There was a significant correlation ( r = 0.529 ) between the amount of IgG bound by the two peptides , but neither of these values correlated well with their P16473 SAb activity nor thyrotropin-binding inhibitor DB00024 receptor antibody ( P16473 IAb ) activity . P16473 SAb inhibiting effects of these peptides were then analysed by measuring P16473 SAb activity after incubation with the peptides . Among eight Graves ' IgGs tested the P16473 SAb activity of three was inhibited by both peptides , two were inhibited only by P354-14 and three were not affected by either . These P16473 SAb inhibiting effects were dose-dependent and reproducible . To confirm these findings , a peptide-sepharose gel affinity absorption study was performed . Eleven Graves ' IgGs were applied to both peptide gels and the P16473 SAb activity of the unabsorbed fraction was measured . The P16473 SAb activity of five IgGs was strongly absorbed only by P354-14 and five others were absorbed by both peptides to an almost similar extent. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . Cytoplasmic anchoring of DB02527 -dependent protein kinase ( PKA ) by A-kinase anchor proteins ( AKAPs ) is required for meiotic arrest of porcine full-grown and growing oocytes . Mammalian growing oocytes ( GOs ) lack the ability to resume meiosis , although the molecular mechanism of this limitation is not fully understood . We previously hypothesized that the meiotic incompetence of porcine GOs was attributed to complex spatial-temporal regulation of DB02527 -dependent protein kinase ( PKA ) by A-kinase anchor proteins ( AKAPs ) , but found that Q92667 is not involved in the meiotic incompetence of porcine GOs . In the present study , we cloned porcine cDNAs of P24588 and AKAP7alpha , and found that inhibiting the expression of these AKAPs induced PKA translocation into the nucleus and promoted meiotic resumption of porcine GOs without affecting the total PKA activity of GOs , whereas overexpressing these AKAPs had no effect . Because AKAPs regulate PKA localization through binding with regulatory subunits of PKA ( PKA-Rs ) , PKA-R binding with AKAPs was inhibited by AKAP-binding inhibition peptides or PKA-R expression inhibition by antisense RNAs . We found that the expression inhibition and binding inhibition of P10644 , an isoform of mammalian PKA-R , promoted meiotic resumption of porcine GOs , whereas these inhibitions of P13861 , another PKA-R isoform , had no effect . In contrast , the expression inhibition and binding inhibition of P13861 had higher effects than those of P10644 on meiotic resumption of porcine full-grown oocytes . These results suggest that cytoplasmic anchoring of PKA by AKAPs is required for meiotic arrest of oocytes and that the PKA-R isoform working for the maintenance of meiotic arrest changed from P10644 to P13861 during the acquisition of meiotic competence . Acute effects of acamprosate and MPEP on ethanol Drinking-in-the-Dark in male C57BL/6J mice . BACKGROUND : Recently , a simple procedure in mice , Drinking-in-the-Dark ( DID ) , was hypothesized to have value for medication development for human alcoholism . In DID , mice are offered intermittent , limited access to ethanol over a series of days during the dark phase that results in rapid drinking to intoxication in predisposed genotypes . METHODS : We measured the effects of acamprosate or MPEP , metabotropic glutamate 5 receptor ( P41594 ) antagonist , on intake of 20 % ethanol , plain tap water or 10 % sugar water using the DID procedure in male C57BL/6J mice . RESULTS : DB00659 ( 100 , 200 , 300 , or 400 mg/kg ) dose dependently decreased ethanol drinking with 300 mg/kg reducing ethanol intake by approximately 20 % without affecting intake of plain water or 10 % sugar water . MPEP ( 1 , 3 , 5 , 10 , 20 , or 40 mg/kg ) was more potent than acamprosate with 20 mg/kg reducing ethanol intake by approximately 20 % and for longer duration without affecting intake of plain water or 10 % sugar water . CONCLUSIONS : These results support the hypothesis that P41594 signaling plays a role in excessive ethanol intake in DID and suggest DID may have value for screening novel compounds that reduce overactive glutamate signaling for potential pharmaceutical treatment of excessive ethanol drinking behavior . In vitro suppression of anti- DB00024 receptor antibody by autologous anti-idiotypic antibody in patients with Graves ' disease . Regulation of anti- DB00024 receptor antibody ( anti- P16473 antibody ) in Graves ' patients ( n = 11 ) by anti-idiotypic antibody was studied using patients sera before and after one year of Methimazole treatment . Patients sera with high level of anti- P16473 antibody ( 110 + 41.9 U/I ) were incubated with pooled control and autologous ( with low level or anti- P16473 antibody negative ) sera containing equimolar IgG G . The mixture was centrifuged and the supernatants were tested for anti- P16473 antibodies ( TRAK , Henning ) . It was found that the autologous sera from patients with remission were able to suppress significantly the titre of anti- P16473 antibodies ( p < 0.001 ) , whereas the controls were capable of a less remarkable inhibition . F(ab')2 fragments of autologous IgG from remission could also suppress the levels of P16473 antibodies . It was concluded that anti-anti- P16473 antibodies in sera of Graves ' patients might be , at least in part , responsible for inducing and maintaining remission and suppression of autoantibodies to P16473 . It is hypothesized that the idiotype system is part of the network of natural autoantibodies and that its perturbation may give rise to pathogenetic antibodies . On the basis of this observation the autologous sera could have therapeutical implication in an accelerated remission of hyperthyroidism in Graves ' disease . Identification and analysis of specific chromosomal region adjacent to exogenous Dhfr-amplified region in Chinese hamster ovary cell genome . Chinese hamster ovary ( CHO ) cells are widely used for the stable production of recombinant proteins . Gene amplification techniques are frequently used to improve of protein production , and the dihydrofolate reductase ( P00374 ) gene amplification system is most widely used in the CHO cell line . We previously constructed a CHO genomic bacterial artificial chromosome ( BAC ) library from a mouse Dhfr-amplified CHO DR1000L-4N cell line and one BAC clone ( Cg0031N14 ) containing the CHO genomic DNA sequence adjacent to Dhfr was selected . To identify the specific chromosomal region adjacent to the exogenous Dhfr-amplified region in the CHO cell genome , we performed further screening of BAC clones to obtain other Dhfr-amplified regions in the CHO genome . From the screening by high-density replica filter hybridization using a digoxigenin-labeled pSV2-dhfr/hGM- P04141 probe , we obtained 8 new BAC clones containing a Dhfr-amplified region . To define the structures of the 8 BAC clones , Southern blot analysis , BAC end sequencing and fluorescence in situ hybridization ( Q5TCZ1 ) were performed . These results revealed that all the selected BAC clones contained a large palindrome structure with a small inverted repeat in the junction region . This suggests that the obtained amplicon structure in the Dhfr-amplified region in the CHO genome plays an important role in exogenous gene amplification . 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 . Expression of endometrial protein kinase a during early pregnancy in bonnet monkeys ( Macaca radiata ) . Embryo-induced signaling pathways are considered to be important for initiation and sustenance of pregnancy . However many of these pathways remain to be deciphered in primates . In the present study , differential display RT-PCR was used to identify genes or gene fragments that are differentially expressed in endometrium of bonnet monkeys ( Macaca radiata ) on Day 6 of pregnancy . Of several fragments found to be differentially expressed , a fragment of 567 base pair ( named GG1 ) was characterized in detail . GG1 was highly represented in endometrium of pregnant animals compared with that of nonpregnant animals . Sequencing analysis revealed homology of this fragment to exons 7 , 8 , 9 , and 10 and surprisingly to intron 6 of DB02527 -dependent protein kinase A ( PKA ) regulatory type I alpha ( tissue-specific extinguisher 1 ) ( P10644 ) . The increased expression of this fragment in gestational endometrium was confirmed by quantitative PCR studies . Two transcripts of 3.0 kilobase ( kb ) and 1.5 kb were detected in Northern blot probed with labeled GG1 . Protein expressions of alpha regulatory ( P10644 ) and alpha catalytic ( P17252 ) subunits of PKA were also higher in gestational endometrium compared with that in nongestational endometrium . Further in vitro studies using human endometrial explants demonstrated regulation of P10644 ( or GG1 ) and prostaglandin-endoperoxide synthase 2 or cyclooxygenase 2 ( P35354 ) by estradiol . This is the first study to date on the differential expression of PKA in primate endometrium during early pregnancy and its in vitro regulation by estradiol . 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 . Mapping of the regulatory type I alpha and catalytic beta subunits of DB02527 -dependent protein kinase and interleukin 1 alpha and 1 beta in the pig . The genes coding for the regulatory type I alpha subunit ( P10644 ) and the catalytic beta subunit ( P22694 ) of DB02527 -dependent protein kinase and the genes for interleukin 1 alpha ( P01583 ) and interleukin 1 beta ( P01584 ) were localized in the pig by means of radioactive in situ hybridization . P10644 was mapped to 12p1.4 and PRKARB to 6q3.1 --> q3.3 . The genes for P01583 and P01584 were both assigned to Chromosome ( Chr ) 3 , in the region q1.2 --> q1.3 and q1.1 --> q1.4 , respectively . The cDNA nucleotide sequences of these porcine genes were compared with those of human , mouse , and cattle . The location of the genes was discussed in relation to the position of their homologous loci in these mammalian species . DB01892 , an Anti-Inflammatory Constituent from St . John 's Wort , Inhibits Microsomal Prostaglandin E(2) Synthase-1 and Suppresses Prostaglandin E(2) Formation in vivo . The acylphloroglucinol hyperforin ( Hyp ) from St . John 's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of P09917 . Here , we investigated whether Hyp also interferes with prostanoid generation in biological systems , particularly with key enzymes participating in prostaglandin (PG)E(2) biosynthesis , i.e. , cyclooxygenases ( P36551 ) -1/2 and microsomal PGE(2) synthase ( mPGES ) -1 which play key roles in inflammation and tumorigenesis . Similar to the mPGES-1 inhibitors MK-886 and MD-52 , Hyp significantly suppressed PGE(2) formation in whole blood assays starting at 0.03-1 μM , whereas the concomitant generation of P36551 -derived 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid , thromboxane B(2) , and 6-keto P49763 (1α) was not significantly suppressed up to 30 μM . In cell-free assays , Hyp efficiently blocked the conversion of PGH(2) to PGE(2) mediated by mPGES-1 ( IC(50) = 1 μM ) , and isolated P36551 enzymes were not ( P35354 ) or hardly ( P23219 ) suppressed . Intraperitoneal ( i.p. ) administration of Hyp ( 4 mg kg(-1) ) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels , and Hyp ( given i.p. ) inhibited carrageenan-induced mouse paw edema formation ( ED(50) = 1 mg kg(-1) ) being superior over indomethacin ( ED(50) = 5 mg kg(-1) ) . We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp . 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 . Mechanism of activation and functional role of protein kinase Ceta in human platelets . The novel class of protein kinase C ( nPKC ) isoform eta is expressed in platelets , but not much is known about its activation and function . In this study , we investigated the mechanism of activation and functional implications of nPKCeta using pharmacological and gene knock-out approaches . nPKCeta was phosphorylated ( at DB00156 -512 ) in a time- and concentration-dependent manner by 2MeSADP . Pretreatment of platelets with P59665 -2179 , a P47900 receptor antagonist , or YM-254890 , a G(q) blocker , abolished 2MeSADP-induced phosphorylation of nPKCeta . Similarly , ADP failed to activate nPKCeta in platelets isolated from P47900 and G(q) knock-out mice . However , pretreatment of platelets with Q9H244 receptor antagonist , AR-C69331MX did not interfere with ADP-induced nPKCeta phosphorylation . In addition , when platelets were activated with 2MeSADP under stirring conditions , although nPKCeta was phosphorylated within 30 s by ADP receptors , it was also dephosphorylated by activated integrin alpha(IIb)beta3 mediated outside-in signaling . Moreover , in the presence of SC-57101 , a alpha(IIb)beta3 receptor antagonist , nPKCeta dephosphorylation was inhibited . Furthermore , in murine platelets lacking PP1cgamma , a catalytic subunit of serine/threonine phosphatase , alpha(IIb)beta3 failed to dephosphorylate nPKCeta . Thus , we conclude that ADP activates nPKCeta via P47900 receptor and is subsequently dephosphorylated by PP1gamma phosphatase activated by alpha(IIb)beta3 integrin . In addition , pretreatment of platelets with eta-RACK antagonistic peptides , a specific inhibitor of nPKCeta , inhibited ADP-induced thromboxane generation . However , these peptides had no affect on ADP-induced aggregation when thromboxane generation was blocked . In summary , nPKCeta positively regulates agonist-induced thromboxane generation with no effects on platelet aggregation . Chronic delivery of a thrombospondin-1 mimetic decreases skeletal muscle capillarity in mice . Angiogenesis is an essential process for normal skeletal muscle function . There is a growing body of evidence suggesting that thrombospondin-1 ( P07996 -1 ) , a potent antiangiogenic protein in tumorigenesis , is an important regulator of both physiological and pathological skeletal muscle angiogenesis . We tested the hypothesis that chronic exposure to a P07996 -1 mimetic ( DB05434 ) , which targets the P16671 P07996 -1 receptor , would decrease skeletal muscle capillarity as well as alter the balance between positive and negative angiogenic proteins under basal conditions . Osmotic minipumps with either DB05434 or vehicle ( 5 % dextrose ) were implanted subcutaneously in the subscapular region of C57/BL6 mice for 14 days . When compared to the vehicle treated mice , the DB05434 group had a 20 % decrease in capillarity in the superficial region of the gastrocnemius ( GA ) , 11 % decrease in the plantaris ( Q02083 ) , and a 35 % decrease in the soleus ( SOL ) . DB05434 also decreased muscle protein expression of vascular endothelial growth factor ( P15692 ) in both the GA ( -140 % ) and SOL ( -62 % ) ; however there was no change in P15692 in the Q02083 . Serum P15692 was not altered in DB05434 treated animals . Endogenous P07996 -1 protein expression in all muscles remained unaltered . Tunnel staining revealed no difference in muscle apoptosis between DB05434 and vehicle treated groups . These data provide evidence that the anti-angiogenic effects of P07996 -1 are mediated , at least in part , via the P16671 receptor . It also suggests that under physiologic conditions the P07996 -1/ P16671 axis plays a role in regulating basal skeletal muscle microvessel density . Selective peroxisome proliferator-activated receptor α modulators ( SPPARMα ) : the next generation of peroxisome proliferator-activated receptor α-agonists . Dyslipidemia is a major risk factor for cardiovascular ( CV ) disease - the primary cause of death , worldwide . Although reducing levels of low-density lipoprotein-cholesterol can significantly reduce CV risk , a high level of residual risk persists , especially in people with obesity-related conditions , such as metabolic syndrome and type 2 diabetes mellitus . Q07869 - ( PPARα- ) agonists ( e.g. fibrates ) , play a central role in the reduction of macro- and microvascular risk in these patients . However , the currently available fibrates are weak ( PPARα-agonists ) with limited efficacy due to dose-related adverse effects . To address this problem , a new generation of highly potent and selective PPARα-modulators ( SPPARMα ) is being developed that separate the benefits of the PPARα-agonists from their unwanted side effects . Among these , aleglitazar ( a dual PPARα/γ agonist ) and DB05187 ( a dual Q07869 α/δ agonist ) have recently entered late-phase development . Although both compounds are more potent PPARα-activators than fenofibrate in vitro , only aleglitezar is more effective in lowering triglycerides and raising high-density lipoprotein-cholesterol ( HDL-C ) in humans . However , it is also associated with a potential risk of adverse effects . More recently , a highly potent , specific PPARα-agonist ( K-877 ) has emerged with SPPARMα characteristics . Compared to fenofibrate , K-877 has more potent PPARα-activating efficacy in vitro , greater effects on triglycerides- and HDL-C levels in humans , and a reduced risk of adverse effects . If successful , K-877 has the potential to supersede the fibrates as the treatment of choice for patients with residual CV risk associated with metabolic syndrome and type 2 diabetes . 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 . Absence of P05112 , and not suppression of the Th2 response , prevents development of experimental autoimmune Graves ' disease . In autoimmune Graves ' disease ( GD ) , autoantibodies bind to the thyrotropin receptor ( P16473 ) and cause hyperthyroidism . We studied the effects of fms-like tyrosine kinase receptor 3 ligand ( Flt3-L ) or GM- P04141 treatment on the development of experimental autoimmune GD ( EAGD ) in mice , a slowly progressing Ab-mediated organ-specific autoimmune disease of the thyroid induced by immunization with syngeneic cells expressing P16473 . Flt3-L and GM- P04141 treatment resulted in up-regulation of CD8a(+) and CD8a(-) dendritic cells , and skewing of cytokine and immune responses to P16473 in favor of Th1 and Th2 , respectively . However , this skewing did not persist until the later stages , and thus failed to affect the course or severity of the disease . To determine whether the total absence of either P05112 or P01579 could affect the development of EAGD , we immunized wild-type , P01579 (-/-) and P05112 (-/-) BALB/c mice with P16473 . Nearly 100 % of the wild-type and P01579 (-/-) mice developed EAGD with optimal P16473 -specific immune responses , while P05112 (-/-) mice completely resisted disease and showed delayed and suboptimal pathogenic Ab response . These data demonstrated that skewing immune responses to P16473 , using either Flt3-L or GM- P04141 , in favor of Th1 or Th2 , respectively , may not be sufficient to alter the course of the disease , while the complete absence of P05112 , but not P01579 , can prevent the development of EAGD .

MH_train_1561

MH_train_1561

MH_train_1561

interacts_with DB00682?

multiple_choice

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 . Hidden risk genes with high-order intragenic epistasis in Alzheimer 's disease . Meta-analysis of data from genome-wide association studies ( GWAS ) of Alzheimer 's disease ( AD ) has confirmed the high risk of P02649 and identified twenty other risk genes/loci with moderate effect size . However , many more risk genes/loci remain to be discovered to account for the missing heritability . The contributions from individual singe-nucleotide polymorphisms ( SNPs ) have been thoroughly examined in traditional GWAS data analysis , while SNP-SNP interactions can be explored by a variety of alternative approaches . Here we applied generalized multifactor dimensionality reduction to the re-analysis of four publicly available GWAS datasets for AD . When considering 4-order intragenic SNP interactions , we observed high consistency of discovered potential risk genes among the four independent GWAS datasets . Ten potential risk genes were observed across all four datasets , including P54750 , Q15413 , Q02763 , Q9BQT8 , LOC729852 , Q8IZU9 , P54829 , P23945 , O60260 , and P08235 . These potential risk genes discovered by generalized multifactor dimensionality reduction are highly relevant to AD pathogenesis based on multiple layers of evidence . The genetic contributions of these genes warrant further confirmation in other independent GWAS datasets for AD . HER-2 peptides p776 and P08709 , N-terminal-linked with Ii-Key tetramer ( LRMK ) help the proliferation of E75-TCR+ cells : The dependency of help on the side chains of LRMK-extended peptide pointed towards the T cell receptor . The objective of this study was to determine whether peptides consisting of the Ii-Key peptide LRMK linked to the N-terminal ends of HER-2 peptides would stimulate the expansion of antigen-specific E75-TCR+CD8+ cells . The peptides tested were N-acetylated and linked to an alpha-amide at the C-terminus ; some of the peptides contained epsilon-aminovaleric acid ( Ava ) between the LRMK and the HER-2 peptide . Of the seven LRMK-HER-2 peptides tested to date , three effectively induced P01579 production by peripheral blood mononuclear cells ( PBMCs ) from healthy donors and women with ductal carcinoma in situ . A fusion peptide , LRMK-Ava-HER-2(777-789) , was more immunogenic than the natural HER-2(777-789) antigen , G89 , with regard to P01579 production . In combination with the CD8-activating peptide E75 [ HER-2(369-377) ] LRMK-p776 and LRMK-Ava- P08709 induced the proliferation of E75-TCR(Med+Hi) CD8+ cells to a greater extent than did 1,000 or 5,000 nM of E75 alone , respectively . The induction effects were strongest at 600 nM for LRMK-p776 and 3,000 nM for LRMK-Ava- P08709 . At 3,000 nM , LRMK-p776 was cytotoxic to PBMCs . LRMK-p776 and P08709 had a similar specificity and preferences for binding HLA-DR molecules . The molecular modeling of HLA-DR:LRMK-p776 and HLA-DR:LRMK-Ava- P08709 complexes revealed the side chains of the peptides , which pointed towards the T-cell receptor . Differences in side chain orientation introduced by various N-terminal extensions of MHC class II-bound peptides should be important for directing P01730 + cells to stimulate CD8+ cells or for eliminating regulatory T cells in cancer immunotherapy . 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 . Can P35354 inhibitor-induced increase in cardiovascular disease risk be modified by essential fatty acids ? Selective P35354 inhibitors increase the risk of myocardial infarction and stroke . This has been attributed to their ability to inhibit endothelial P35354 derived prostacyclin ( DB01240 ) but not platelet P23219 derived thromboxane A2 ( TXA2 ) . On the other hand , aspirin blocks both P23219 and P35354 enzymes without decreasing DB01240 but blocks TXA2 synthesis that explains its beneficial action in the prevention of coronary heart disease ( Q8NE62 ) . The inhibitory action of aspirin on P23219 and P35354 enzymes enhances the tissue concentrations of dihomo-gamma-linolenic acid ( DB00154 ) , arachidonic acid , eicosapentaenoic acid ( EPA ) , and docosahexaenoic acid ( DB01708 ) . These fatty acids form precursors to PGE1 , DB01240 , PGI3 , lipoxins ( LXs ) , and resolvins that have anti-inflammatory actions . In contrast , increase in the concentrations of DB00154 , AA , EPA , and DB01708 is much less with specific P35354 inhibitors since they do not block the formation of eicosanoids through P23219 pathway . P35354 inhibitors interfere with the formation of LXs and resolvins that have neuroprotective and cardioprotective actions . EPA and DB01240 have anti-arrhythmic action . EPA , DB01708 , and AA augment eNO formation that prevents atherosclerosis . This suggests that P35354 inhibitors increase cardiovascular and stroke risk by interfering with the formation of eNO , DB01240 , LXs , and resolvins and implies that combining EFAs with P35354 inhibitors could prevent these complications . 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 . 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 . Modulation of superantigen-induced T-cell deletion by antibody anti-Pgp-1 ( P16070 ) . We examined the effects of anti-Pgp-1 ( P16070 ) antibody on the in vitro deletion of murine P01730 and CD8 single positive T cells induced by Staphylococcal enterotoxin B ( Q9Y6X0 ) . Soluble anti-Pgp-1 antibody enhanced the apoptosis and decreased the proliferation of Q9Y6X0 -responding T cells . In contrast , cross-linked anti-Pgp-1 antibody provided costimulatory signals for the T-cell activation induced by anti-CD3 antibody . DB08818 ( HA ) , a ligand of Pgp-1 , did not affect proliferation and deletion induced by Q9Y6X0 , whereas it mimicked the effects of the cross-linked antibody in anti-CD3-driven proliferation . T-cell Pgp-1 surface expression after 48 hr incubation with Q9Y6X0 was unchanged as compared to unstimulated cells . However , when the memory T cells were established , some V beta 8+ ( Q9Y6X0 -specific ) T cells Pgp-1low became Pgp-1high , displaying a bimodal character . Moreover , the Pgp-1 increased expression correlated with an increase of Pgp-1 soluble form in the supernatant . These findings suggested that signals following the triggering of the Pgp-1 molecule are important in controlling T-cell survival . Generation and characterization of a human monoclonal IgM antibody that recognizes a conserved epitope shared by lipopolysaccharides of different gram-negative bacteria . A hybridoma cell line secreting a human monoclonal antibody ( humab ) directed to an epitope in the lipid A region of lipopolysaccharides of Gram-negative bacteria was isolated . Peripheral blood lymphocytes ( PBL ) obtained from a healthy volunteer were immortalized by Epstein-Barr virus ( EBV ) transformation . Lymphoblastoid cell lines ( LCL ) secreting antibodies to the lipopolysaccharides of Gram-negative bacteria were determined by an enzyme-linked immunosorbent assay ( ELISA ) and subsequently fused with the human-mouse heteromyeloma cell line CB- P08709 by polyethylenglycol ( PEG ) -mediated fusion . A hybridoma line producing a humab ( LPD5H4 ) , of the IgM/lambda isotype , which strongly reacted with the lipid A portion of Salmonella and E. coli spp. in ELISA , was established . The antibody was purified by hydrophobic interaction chromatography and gel filtration . Immunoblotting experiments showed a strong reactivity of the humab LPD5H4 with the lower molecular species of different rough and smooth lipopolysaccharide ( LPS ) types of the bacteria species Salmonella , E. coli , Klebsiella , and Neisseria meningitidis , whereas those of Pseudomonas spp. were negative . Binding of humab LPD5H4 to solid phase bound lipid A and different rough mutants of LPS could be inhibited by the corresponding antigens in solution . Competition assays with a murine monoclonal antibody to lipid A and with polymyxin B indicate that humab LPD5H4 recognizes its epitope in this extremely conserved part of the LPS molecule . In vitro tests demonstrated that the MAb is able to partially inhibit the LPS-induced release of P01375 using isolated peripheral blood mononuclear cells ( PBMC ) . DB01404 saponin metabolite induces apoptosis in MCF-7 breast cancer cells through the modulation of AMP-activated protein kinase . Previous studies have shown that the ginseng saponin metabolite , Compound K ( 20-O-d-glucopyranosyl-20(S)-protopanaxadiol , IH901 ) , suppresses proliferation of various cancers and induces apoptosis . AMP-activated protein kinase ( AMPK ) is a sensor of cellular energy states and is involved in apoptosis of cancer cells . We hypothesized that Compound K may exert cytotoxicity in MCF-7 human breast cancer cells through modulation of AMPK , followed by a decrease in cyclooxygenase-2 ( P35354 ) expression . Compound K inhibited cell growth , induced apoptosis via generation of reactive oxygen species ( ROS ) , as well as decreasing P35354 expression and prostaglandin E(2) ( PGE(2) ) levels . These effects of Compound K were induced via an AMPK-dependent pathway and were abrogated by a specific AMPK inhibitor . These results suggest that Compound K induced apoptosis by modulating AMPK- P35354 signaling in MCF-7 human breast cancer cells . 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 . 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 . 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 . Comparison of rating scales used to evaluate DB01235 -induced dyskinesia in the 6-OHDA lesioned rat . Abnormal involuntary movement ( AIM ) rating scales are frequently used to study the mechanisms underlying DB01235 -induced dyskinesia ( LID ) in 6-OHDA lesioned rodents and the propensity of novel treatments for Parkinson 's disease to induce or alleviate similar abnormal behaviours . Despite the existence of at least one well validated method , other AIM scales are also in use . Moreover , there have been developments and variations in the original scales and their methods of use , without re-validation . In this study , 6-OHDA medial forebrain bundle lesioned Sprague-Dawley rats were treated with chronic DB01235 6 mg/kg/day for 5 weeks followed by 12 mg/kg/day for another 5 weeks . Rats were assessed weekly by simultaneous ratings on four published AIM and stereotypy scales with concurrent recording of rotation , over 3 hours following DB01235 injection . Three contemporary AIM scales have then been validated pharmacologically using agents that are known to reduce LID clinically and in primates ( amantadine ) or to interfere with the activity of DB01235 ( the D(1) and P14416 antagonists , P35240 -23390 and raclopride ) respectively . We also demonstrate that AIM , stereotypic and rotational behaviour are distinct motor dysfunctions induced by chronic and acute treatment of DB01235 , and should be assessed separately . The undertaking of assessments at multiple time points is essential especially when testing the efficacy of new potential anti-dyskinetic treatments . Importantly critical to all AIM and rotation testing is the internal validation of both the scale being used and the environment being used . 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 . Molecular characteristics of the N-terminal region of the quail follitropin receptor . We determined the primary structure of follitropin receptor ( P23945 ) at its N-terminal extracellular domain , which is the key region of specific hormone binding in avian ( quail ) species . In this region , quail P23945 showed about 70 % homology with mammalian DB00094 -Rs at the level of predicted amino acid sequences . The leucine-rich repetitive motif which is conserved in all mammalian DB00094 -Rs was also detected in the quail P23945 . However , some unique amino acid replacements were found at the positions of cystein residues and potential N-linked glycosylation sites . The sequence of the quail lutropin receptor ( LH-R ) previously defined by us showed a homology between P23945 and LH-R of 47.4 % . This value is close to those between mammalian DB00094 -Rs and LH-Rs , which in the corresponding region are human , 50.3 % ; rat , 50.9 % . O43609 promotes the degradation of Q03405 and inhibits Q03405 -mediated cell adhesion and proliferation . DB00013 plasminogen activator receptor ( Q03405 ) is a P06744 anchored cell surface protein that is closely associated with invasion , migration , and metastasis of cancer cells . Many functional extracellular proteins and transmembrane receptors interact with Q03405 . However , few studies have examined the association of Q03405 with cytoplasm proteins . We previously used yeast two-hybrid screening to isolate several novel Q03405 -interacting cytoplasmic proteins , including Sprouty1 ( O43609 ) , an inhibitor of the ( Ras-mitogen-activated protein kinase ) MAPK pathway . In this study , we show that O43609 interacts with Q03405 and directs it toward lysosomal-mediated degradation . Overexpression of O43609 decreased the cell surface and cytoplasmic Q03405 protein level . Moreover , O43609 overexpression augmented Q03405 -induced cell adhesion to vitronectin as well as proliferation of cancer cells . Our results also further support the critical role of O43609 contribution to tumor growth . In a subcutaneous tumor model , overexpression of O43609 in HCT116 or A549 xenograft in athymic nude mice led to great suppression of tumor growth . These results show that O43609 may affect tumor cell function through direct interaction with Q03405 and promote its lysosomal degradation . 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 . Genetic polymorphisms for the study of multifactorial stroke . Single-gene disorders explain only a minority of stroke cases . Stroke represents a complex trait , which is usually assumed to be polygenic . On this topic , the role of a wide number of candidate genes has been investigated in stroke through association studies , with controversial results . Therefore , it is difficult for the clinician to establish the validity and the level of clinical applicability of the previously reported associations between genetic factors and stroke . This review is an update and an extensive analysis of the more recent association studies conducted in stroke . We evaluated a number of studies on several candidate genes ( including P12259 , F2 , P02671 / P02675 / P02679 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / Q9UKI9 / P04054 / P08514 , P17301 , P07359 , P12821 , AGT , NOS3 , P02649 , P06858 , P27169 , Q08499 , P20292 , P42898 , Q99707 , and P35520 ) , providing a final panel of genes and molecular variants . We categorized this panel in relation to the degree of association with stroke , supported by the results of meta-analyses and case-control studies . Our findings could represent a useful tool to address further molecular investigations and to realize more detailed meta-analyses . 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 .

MH_train_1562

MH_train_1562

MH_train_1562

interacts_with DB06684?

multiple_choice

Complement deposition and microglial activation in the outer retina in light-induced retinopathy : inhibition by a P08908 agonist . PURPOSE : Increasing evidence supports a role for complement in the pathogenesis of age-related macular degeneration ( AMD ) . This study evaluated retinal microglia , T-lymphocytes , and complement deposition in a light-induced retinopathy model . The effect of a serotonin ( 5-hydroxytryptamine , 5-HT(1A) ) agonist on these processes was investigated . METHODS : Rats were dark adapted for 24 hours before a 6-hour blue light exposure . Some animals were predosed subcutaneously with AL-8309A . Retinas were evaluated at different times after light exposure . Paraffin sections were stained with antibody for a microglial marker ( Iba1 ) , a T-lymphocyte marker ( CD3 ) , and complement components C1q , P01024 , factor B , factor H , and membrane attack complex ( MAC ) . RESULTS : Light exposure resulted in substantial photoreceptor and Q96AT9 loss . Robust microglia activation and migration to the outer retina occurred rapidly . Substantial T-lymphocyte recruitment did not occur . Complement alternative pathway was strongly activated , resulting in the deposition of P01024 , factor B , factor H , and MAC in the area of photic lesions . Dosing with AL-8309A prevented retinal lesions and decreased microglia activation/recruitment and complement deposition in the outer retina . CONCLUSIONS : In blue light exposed retinas , microglia were activated and migrated toward the outer retina , whereas a T-lymphocyte response was minimal . The innate immune system was markedly activated , with substantial complement deposition in the outer retina after light exposure . This complement deposition was prevented by AL-8309A . This model may be useful in the evaluation of complement inhibitors and other neuroprotectants intended for ocular use . AL-8309 is under evaluation in the clinic and may be useful in the treatment of AMD . Methylenedioxymethamphetamine induces spontaneous tail-flicks in the rat via P08908 receptors . In rats lightly restrained in horizontal cylinders , (+/-)-3,4-methylenedioxymethamphetamine ( DB01454 ) dose dependently ( 0.16-10.0 mg/kg , s.c. ) elicited spontaneous tail-flicks ; that is , tail-flicks in the absence of extraneous stimulation . In contrast , amphetamine over a similar dose-range was inactive . Selective inhibitors of 5-hydroxytryptamine ( 5-HT ) uptake and carrier-mediated 5-HT release , paroxetine and citalopram , did not induce spontaneous tail-flicks themselves and blocked those induced by DB01454 . In distinction , maprotiline and bupropion , selective inhibitors of noradrenaline and dopamine uptake , respectively , failed to modify the action of DB01454 . Spontaneous tail-flicks elicited by DB01454 were unaffected by the selective 5- Q9H205 receptor antagonists , ICS 205,930 and GR 38032F . They were attenuated by the mixed 5-HT1/5-HT2 receptor antagonist , methiotepin , the mixed P08908 / P28222 receptor antagonist , (-)-alprenolol and the mixed P08908 /5-HT2 receptor antagonist , spiperone , but not by the selective P28335 /5-HT2 receptor antagonists , ritanserin , ICI 169,369 and ketanserin . The novel P08908 receptor antagonists , BMY 7378 and NAN-190 , each abolished DB01454 -evoked spontaneous tail-flicks . Selective D1 , D2 , alpha 1 , alpha 2 , beta 1 and beta 2 antagonists had little influence upon induction of spontaneous tail-flicks by DB01454 . These data indicate that DB01454 evokes spontaneous tail-flicks in the rat via a release of 5-HT which acts at P08908 receptors . Thus , P08908 receptors appear to be involved in the acute functional actions of DB01454 . The G protein-coupled P08908 receptor causes suppression of caspase-3 through MAPK and protein kinase Calpha . The 5-HT(1A) agonist 8-hydroxy-2 ( di-n-propylamino ) tetralin ( 8-OH-DPAT ) causes inhibition of caspase-3 and apoptosis via the extracellular signal-regulated kinases ( P27361 /2 ) in hippocampal P0CJ69 -5 cells . Two 5-HT(1A) agonists , Repinotan hydrochloride ( BAY x 3702 ) and 8-OH-DPAT , block caspase-3 activation and apoptosis caused by anoxia/reoxygenation and H(2)O(2) treatment . This is reversed upon transient expression of dominant negative Ras ( N17Ras ) and P04049 ( Raf301 ) , confirming the involvement of Ras and P04049 in this 5-HT(1A)-R --> P27361 /2 --> caspase-3 pathway . A selective inhibitor of phospholipase Cbeta ( PLCbeta ) ( U73122 ) but not a general protein kinase C ( PKC ) inhibitor ( GFX ) reversed the 5-HT(1A)-R-mediated P27361 /2 stimulation . However , both GFX and the PKCalpha and PKCbeta(1) inhibitor Gö6976 reversed the P27361 /2-mediated inhibition of caspase-3 . P29323 -dependent activation of only PKCalpha was observed in immunoprecipitates obtained from 5-HT(1A) agonist-treated P0CJ69 -5 cells . Finally , transient expression of kinase-negative PKCalpha eliminated the 8-OH-DPAT-evoked block on the H(2)O(2)-triggered caspase-3 stimulation , establishing PKCalpha as a link between P29323 and caspase-3 ( 5-HT(1A)-R --> P98160 --> P27361 /2 --> PKCalpha --> caspase-3 ) . Our results elucidate a novel yet general , neuroprotective pathway through which G protein-coupled receptors could cause inhibition of effector caspases , such as caspase-3 . 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 . 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 . 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 . High doses of intravenous Ig inhibit in vitro uptake of C4 fragments onto sensitized erythrocytes . We have recently reported that intravenous Ig ( DB00028 ) inhibits uptake of activated P01024 fragments onto antibody-sensitized red blood cells ( RBCs ) . To elucidate the mechanism by which DB00028 exerts its effect on the complement system , we examined the possible interference with the C4 step of the classical complement cascade . We examined the capacity of autologous serum containing high concentrations of human DB00028 to deposit C4 fragments onto model targets ( guinea pig and/or human erythrocytes sensitized with rabbit anti-guinea pig/human erythrocytes IgG antibody ) . C4 binding was quantified with radiolabeled anti-C4 . Guinea pig serum with added DB00028 suppressed C4 uptake onto IgG-sensitized guinea pig erythrocytes at all time points ( 0 , 5 , 15 , and 30 minutes ) . Using sera of guinea pigs treated with increasing doses of DB00028 , this effect was shown to be dose-responsive . Serum from a patient treated with DB00028 showed reduced C4 uptake onto sensitized homologous RBCs . In comparison with the serum from the same patient before DB00028 therapy was administered , levels were decreased almost to background . C4 functional titers in those two samples were not different . P01024 uptake was studied in parallel with C4 to compare the degree of inhibition using sera with increasing doses of DB00028 in both the human and guinea pig system . P01024 and C4 inhibition curves completely overlapped . Our findings suggest that DB00028 is an effective inhibitor of deposition of early complement activation products ( C4b , C3b ) onto target surfaces and may indicate interference of DB00028 with multiple sites of complement activation . P05231 -receptor polymorphisms rs12083537 , rs2228145 , and rs4329505 as predictors of response to tocilizumab in rheumatoid arthritis . DB06273 ( TCZ ) , a monoclonal antibody targeting the human interleukin-6-receptor ( IL-6R ) , is indicated for the treatment of rheumatoid arthritis ( RA ) . We examined whether three P08887 single-nucleotide polymorphisms rs12083537 , rs2228145 ( formerly rs8192284 ) , and rs4329505 with previously reported functional effects were associated with clinical response to TCZ in a retrospective study cohort consisting of 79 RA patients . Three months after initiation of TCZ therapy , changes in swollen joint count ( SJC ) and , subordinately , tender joint count ( TJC ) , serum-CRP , DAS28-CRP , and EULAR-response were tested for association with the P08887 -haplotype or genotype . The major allele ( A ) of rs12083537 and the minor allele ( C ) of rs4329505 were associated with a poor SJC response ( P=0.02 and 0.02 , respectively ) . Moreover , the AAC-haplotype ( for rs12083537 , rs2228145 , and rs4329505 , respectively ) was associated with a poor SJC response ( P=0.00004 ) and , with borderline significance , EULAR-response ( P=0.05 ) . These data suggest that genetic variation in P08887 may aid in predicting TCZ therapy outcome in RA patients . 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 . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . 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 . Identification of N-(4-piperidinyl)-4-(2,6-dichlorobenzoylamino)-1H-pyrazole-3-carboxamide ( DB05037 ) , a novel cyclin dependent kinase inhibitor using fragment-based X-ray crystallography and structure based drug design . The application of fragment-based screening techniques to cyclin dependent kinase 2 ( P24941 ) identified multiple ( > 30 ) efficient , synthetically tractable small molecule hits for further optimization . Structure-based design approaches led to the identification of multiple lead series , which retained the key interactions of the initial binding fragments and additionally explored other areas of the DB00171 binding site . The majority of this paper details the structure-guided optimization of indazole ( 6 ) using information gained from multiple ligand- P24941 cocrystal structures . Identification of key binding features for this class of compounds resulted in a series of molecules with low nM affinity for P24941 . Optimisation of cellular activity and characterization of pharmacokinetic properties led to the identification of 33 ( DB05037 ) , which is currently being evaluated in clinical trials for the treatment of human cancers . Effects of a non-selective P36551 inhibitor and selective P35354 inhibitors on contractility of human and porcine ureters in vitro and in vivo . BACKGROUND AND PURPOSE : Anti-inflammatory drugs are used in the treatment of acute renal colic . The aim of this study was to investigate the effects of selective P35354 inhibitors and the non-selective P36551 inhibitor diclofenac on contractility of human and porcine ureters in vitro and in vivo , respectively . P23219 and P35354 receptors were identified in human ureter and kidney . EXPERIMENTAL APPROACH : Human ureter samples were used alongside an in vivo pig model with or without partial ureteral obstruction . P23219 and P35354 receptors were located in human ureters by immunohistochemistry . KEY RESULTS : Diclofenac and valdecoxib significantly decreased the amplitude of electrically-stimulated contractions in human ureters in vitro , the maximal effect ( Vmax ) being 120 and 14 % , respectively . DB00580 was more potent in proximal specimens of human ureter ( EC50=7.3 x 10(-11) M ) than in distal specimens ( EC50=7.4 x 10(-10) M ) , and the Vmax was more marked in distal specimens ( 22.5 % ) than in proximal specimens ( 8.0 % ) in vitro . In the in vivo pig model , parecoxib , when compared to the effect of its solvent , significantly decreased the maximal amplitude of contractions ( Amax ) in non-obstructed ureters but not in obstructed ureters . Diclofenac had no effect on spontaneous contractions of porcine ureter in vivo . P23219 and P35354 receptors were found to be expressed in proximal and distal human ureter and in tubulus epithelia of the kidney . CONCLUSIONS AND IMPLICATIONS : Selective P35354 inhibitors decrease the contractility of non-obstructed , but not obstructed , ureters of the pig in vivo , but have a minimal effect on electrically-induced contractions of human ureters in vitro . Analysis of SNP profiles in patients with major depressive disorder . The present study focused on 91 single-nucleotide polymorphisms ( SNPs ) in 21 candidate genes to find associations with major depressive disorder ( MDD ) . In total , 160 healthy controls and 177 patients with MDD were studied . We applied arrayed primer extension ( P27695 ) based genotyping technology followed by association and haplotype analysis . SNPs in P32238 , P21728 , P14416 , and P28335 genes showed nominally significant associations with MDD . None of these associations remained significant after adjustment for multiple testing . Haplotype analysis revealed P32238 haplotypes to be associated with MDD ( global p=0.004 ) . More precisely , we found the GAGT haplotype to be associated with increased risk for MDD ( OR 7.42 , 95 % CI 2.13-25.85 , p=0.002 ) . This haplotype effect remained significant after Bonferroni correction ( p=0.04 after Bonferroni 's adjustment ) . Altogether we were able to find some nominal associations , but due to small sample size these results should be taken as exploratory . However , the effect of GAGT haplotype on the P32238 gene may be considered as increasing the risk for MDD . Systemic delivery and pre-clinical evaluation of nanoparticles containing antisense oligonucleotides and siRNAs . By virtue of their potential to selectively silence oncogenic molecules in cancer cells , antisense oligonucleotides ( ASO ) and small interfering RNAs ( siRNAs ) are powerful tools for development of tailored anti-cancer drugs . The clinical benefit of ASO/siRNA therapeutic is , however , hampered due to poor pharmacokinetics and biodistribution , and suboptimal suppression of the target in tumor tissues . P04049 protein serine/threonine kinase is a druggable signaling molecule in cancer therapy . Our laboratory has developed cationic liposomes for systemic delivery of raf ASO ( DB04973 ) and raf siRNA ( LErafsiRNA ) to human tumor xenografts grown in athymic mice . DB04973 is also the first ASO containing liposomal drug tested in humans . In this article , we primarily focus on a modified formulation of systemically delivered cationic liposomes containing raf antisense oligonucleotide ( md- DB04973 ) . The cationic liposomes were prepared using dimyristoyl 1,2-diacyl-3-trimethylammonium-propane ( DMTAP ) , phosphatidylcholine ( PC ) , and cholesterol ( CHOL ) . The toxicology , pharmacokinetics , biodistribution , target selectivity , and anti-tumor efficacy studies of md- DB04973 were conducted in mice . We demonstrate that md- DB04973 is the next generation of systemically delivered and well-tolerated antisense therapeutic suitable for clinical evaluation . 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 . 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 . Immunohistochemical localization of adherens junction components in blood-brain barrier microvessels of the rat . The morphology and molecular composition of intercellular adherens junctions have most frequently been described in epithelial cells and the fascia adhaerens of the intercalated disc . A group of cytoplasmic molecules is known to be associated with adherens junctions . The intercellular bond is mediated by cadherins which bridge the cells by homophilic binding . Recently , endothelial cells have also been shown to form intercellular junctions of the adherens-type . However , they are morphologically less distinct and little is known about their molecular components . In this study we report the localization of some adherens junction components in intact microvessels of the blood-brain barrier in the rat . We used antibodies raised against alpha-actinin , vinculin , zyxin , cadherin ( antipan-cadherin antibody ) and A- P62158 ( P19022 ) in immunohistochemical experiments at light and electron microscopical levels . Microvessel walls reacted positively for all antigens throughout postnatal development . All antigens were localised , though not necessarily exclusively , to interendothelial junctions . At the ultrastructural level , pan-cadherin reactivity was present throughout the entire length of the cleft . These results could mean that in blood-brain barrier endothelial cells the complex tight junction is embedded in an adherens junction which occupies the entire length of the cleft . [ Changes of protein expression in HepG2 cells with P24941 RNA interference ] . OBJECTIVE : To investigate the effects of stable transfection of P24941 siRNA on biological activities and nuclear proteins of human hepatocellular carcinoma HepG2 cells . METHODS : HepG2 cells were transfected with the eukaryotic expression vector of P(Genesil-1- P24941 ) ; via RNA interference and selected for the ones with stable transfection . We observed the changes in the cell growth curve and cell cycle . The mRNA contents of P24941 and differentially expressed nucleoproteins were detected and analyzed by RT-PCR and two-dimensional ( 2D ) electrophoresis-mass spectrum ( MS ) -database , respectively . Western blotting were used to confirm the differential protein expressions . RESULTS : Compared with P(HK-siRNA);-HepG2 and untransfected groups , the proliferation of HepG2 cells in P( P24941 -siRNA);-HepG2 group was significantly inhibited ( P < 0.01 ) , and the expression of P24941 mRNA significantly decreased in P( P24941 -siRNA);-HepG2 group . Four proteins not expressed in P( P24941 -siRNA);-HepG2 cells were detected by 2D electrophoresis-MS , and they were further confirmed by Western blotting . CONCLUSION : P24941 siRNA significantly suppressed P24941 mRNA expression and the proliferation of HepG2 cells , four proteins not expressed in p( P24941 -siRNA);-HepG2 cells are similar to ribosomal protein P28222 , β-actin , zine finger 276 and chaperonin 10 related protein . Inhibition of noradrenaline release via presynaptic P28222 receptors of the rat vena cava . In the rat inferior vena cava preincubated with 3H-noradrenaline , the effects of nine serotonin ( 5-HT ) receptor agonists and of eight antagonists ( including two beta-adrenoceptor blocking agents ) on the electrically evoked 3H overflow were determined . 1 . 5-HT , 5-carboxamido-tryptamine , 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole ( RU 24969 ) , 5-methoxytryptamine , N,N-dimethyl-5HT , tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow . The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for P28222 binding sites , but not with their affinities for P08908 , P28335 or 5-HT2 binding sites . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 receptor agonist , and ipsapirone , a partial agonist at these receptors , did not inhibit overflow . 2 . Cyanopindolol facilitated the evoked 3H overflow , an effect which was abolished by propranolol . The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol . 3 . The concentration-response curve for 5-HT was shifted to the right by metitepine , metergoline , quipazine , 6-chloro-2-(1-piperazinyl)pyrazine ( MK 212 ) and propranolol which , given alone , did not affect 3H overflow . The apparent pA2 values of these antagonists tended to be correlated with their affinities for P28222 ( but not P08908 , P28335 or 5-HT2 ) binding sites . Ketanserin , a 5-HT2 receptor antagonist , and spiperone , which blocks 5-HT2 and P08908 but not P28222 or P28335 receptors , failed to antagonize the effect of 5-HT. ( ABSTRACT TRUNCATED AT 250 WORDS )

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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 . 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 . 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 . P09429 is involved in autophagy inhibition caused by P37840 /α-synuclein overexpression : a process modulated by the natural autophagy inducer corynoxine B . P37840 /α-synuclein and its rare mutations are considered as the culprit proteins in Parkinson disease ( PD ) . Wild-type ( WT ) P37840 has been shown to impair macroautophagy in mammalian cells and in transgenic mice . In this study , we monitored the dynamic changes in autophagy process and confirmed that overexpression of both WT and P37840 (A53T) inhibits autophagy in PC12 cells in a time-dependent manner . Furthermore , we showed that P37840 binds to both cytosolic and nuclear high mobility group box 1 ( P09429 ) , impairs the cytosolic translocation of P09429 , blocks P09429 - Q14457 binding , and strengthens Q14457 - P10415 binding . Deregulation of these molecular events by P37840 overexpression leads to autophagy inhibition . Overexpression of Q14457 restores autophagy and promotes the clearance of P37840 . siRNA knockdown of Hmgb1 inhibits basal autophagy and abolishes the inhibitory effect of P37840 on autophagy while overexpression of P09429 restores autophagy . Corynoxine B , a natural autophagy inducer , restores the deficient cytosolic translocation of P09429 and autophagy in cells overexpressing P37840 , which may be attributed to its ability to block P37840 - P09429 interaction . Based on these findings , we propose that P37840 -induced impairment of autophagy occurs , in part , through P09429 , which may provide a potential therapeutic target for PD . 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 . 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 . 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 . 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 . 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 . P35354 inhibitors suppress lung cancer cell growth by inducing P38936 via P35354 independent signals . P35354 has been implicated in the control of human non-small cell lung carcinoma ( NSCLC ) cell growth . The mechanisms by which P35354 exerts its mitogenic effects have not been entirely elucidated , but stimulation of prostaglandin E2 production and alterations in the expression of the cyclin-dependent kinase inhibitor P38936 (WAF-1/CIP1/ P38936 )(p2i) have been suggested . Here , we demonstrate that two P35354 inhibitors ( NS398 and DB04743 ) inhibit proliferation and induce apoptosis in NSCLC cells , and these effects were associated with induction of P38936 mRNA and protein expression . However , the anti-growth effect of the P35354 inhibitors and their ability to induce P38936 were not affected by P35354 siRNA suggesting that their actions were P35354 independent . Instead , activation of the MEK-1/Erk pathway was necessary since P35354 inhibitors stimulated the phosphorylation of ERKs , and their effects were blocked by PD98095 , an inhibitor of this pathway . Furthermore , we show that both NS398 and DB04743 induced P38936 gene promoter activity and this was prevented by PD98095 . P35354 inhibitors increased nuclear protein binding to the Spl site in the promoter region of the P38936 gene . Consistent with a role for P38936 , we found that P38936 antisense oligonucleotides prevented the effects of P35354 inhibitors on cell growth . In summary , our results suggest that P35354 inhibitors suppress NSCLC cell growth by inducing the expression of the P38936 gene through MEK-1/ P29323 signaling and DNA-protein interactions involving Spl . These observations unveil a mechanism for P38936 gene regulation by P35354 inhibitors in lung carcinoma cell growth and this pathway represents a potential target for therapy . Protective effect of hyaluronic acid on interleukin-1-induced deregulation of beta1-integrin and insulin-like growth factor-I receptor signaling and collagen biosynthesis in cultured human chondrocytes . The mechanism of protective action of hyaluronic acid ( HA ) on collagen metabolism disturbances in tissues during inflammation is not known . P01308 -like growth factor-I ( P05019 ) receptor and beta1-integrin receptor signaling plays an important role in the regulation of collagen biosynthesis at both transcriptional and post-transcriptional levels . The present study was undertaken to evaluate the effect of IL-1beta ( inductor of experimental inflammation ) on the signaling pathways as well as on collagen biosynthesis , gelatinases and prolidase activity in cultured human chondrocytes and the effect of HA on these processes . It was found that IL-1beta-dependent inhibition of collagen biosynthesis was accompanied by increase in beta1-integrin receptor , NF-kB expressions , and increase in phosphorylation of Q05397 , that resulted in stimulation of metalloproteinase P08253 and P14780 activities , but not prolidase activity and expression . Simultaneously , decrease in expression of P08069 and phosphorylation of Akt and p38 were found . All those processes were counteracted by HA . This suggests that cross talk between beta1-integrin and P05019 receptors is disturbed by IL-1beta , and HA recovers their proper signaling in cultured chondrocytes . We propose that P08069 and beta1-integrin signaling may play an important role in protective effect of hyaluronic acid on interleukin-1-induced inhibition of collagen biosynthesis in cultured human chondrocytes . The role of the Q9BXA5 ligand succinate in hematopoiesis . Regulation of cellular metabolism by the citric acid cycle occurs in the mitochondria . However , the citric acid cycle intermediate succinate was shown recently to be a ligand for the G-protein-coupled receptor Q9BXA5 . Here , we describe a role for succinate and its receptor in the stimulation of hematopoietic progenitor cell ( HPC ) growth . Q9BXA5 mRNA and protein expression were detected in human bone marrow P28906 + progenitor cells , as well as in erythroid and megakaryocyte cultures and the erythroleukemic cell line TF-1 . Treatment of these cell cultures with succinate resulted in increased proliferation rates . The proliferation response of TF-1 cells was pertussis toxin ( PTX ) -sensitive , suggesting a role for Gi signaling . Proliferation was also blocked when TF-1 cells were transfected with small interfering RNA specific for Q9BXA5 . DB00139 stimulated activation of the Erk MAPK pathway and inositol phosphate accumulation in a PTX-sensitive manner . Pretreatment of TF-1 cells with the Erk1/2 kinase ( MEK ) inhibitor PD98059 blocked the proliferation response . DB00139 treatment additionally protected TF-1 cells from cell death induced by serum deprivation . Finally , in vivo administration of succinate was found to elevate the levels of hemoglobin , platelets , and neutrophils in a mouse model of chemotherapy-induced myelosuppression . These results suggest that succinate- Q9BXA5 signaling is capable of promoting HPC development . Diabetes mellitus in cancer patients treated with combination interleukin 2 and alpha-interferon . Diabetes mellitus is thought to be an autoimmune disease caused by destruction of beta cells in pancreatic islets . P01308 resistance in the peripheral tissues may also play a role . Both interleukin 2 ( P60568 ) and alpha interferon can enhance immune function by stimulating formation of cytolytic T cells and/or antigen expression on both normal and tumor cells . This report describes three patients with advanced malignancy who were treated with combination P60568 and alpha interferon who had the onset or worsening of diabetes mellitus . One patient died as a result . There is evidence that interferon can increase insulin resistance and it is likely that both agents can initiate or enhance an ongoing autoimmune process . Physicians using this combination of drugs should be aware of this potential serious toxicity . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . Risk stratification of intermediate-risk acute myeloid leukemia : integrative analysis of a multitude of gene mutation and gene expression markers . Numerous molecular markers have been recently discovered as potential prognostic factors in acute myeloid leukemia ( AML ) . It has become of critical importance to thoroughly evaluate their interrelationships and relative prognostic importance . Gene expression profiling was conducted in a well-characterized cohort of 439 AML patients ( age < 60 years ) to determine expression levels of EVI1 , P19544 , P10415 , P08183 , Q8WXS3 , P36888 , P28906 , P14902 , ERG and Q10571 . A variety of AML-specific mutations were evaluated , that is , P36888 , P06748 , N- DB01367 , K- DB01367 , O75874 , P48735 , and P49715 (DM/SM) ( double/single ) . Univariable survival analysis shows that ( 1 ) patients with P36888 (ITD) mutations have inferior overall survival ( OS ) and event-free survival ( O43281 ) , whereas P49715 (DM) and P06748 mutations indicate favorable OS and O43281 in intermediate-risk AML , and ( 2 ) high transcript levels of Q8WXS3 , P28906 , Q10571 , EVl1 , and ERG predict inferior OS and O43281 . In multivariable survival analysis , P28906 , ERG , and P49715 (DM) remain significant . Using survival tree and regression methodologies , we show that P49715 (DM) , P28906 , and P48735 mutations are capable of separating the intermediate group into 2 AML subgroups with highly distinctive survival characteristics ( OS at 60 months : 51.9 % vs 14.9 % ) . The integrated statistical approach demonstrates that from the multitude of biomarkers a greatly condensed subset can be selected for improved stratification of intermediate-risk AML . Heteromerization of ligand binding domains of N-methyl-D-aspartate receptor requires both coagonists , L-glutamate and glycine . DB01221 receptors ( NMDAR ) are voltage- and glutamate-gated heteromeric ion channels found at excitatory neuronal synapses , the functions of which are to mediate the mechanisms of brain plasticity and , thereby , its higher order functions . In addition to DB00142 , the activation of these heteromeric receptors requires DB00145 or d- DB00133 as a coagonist . However , it is not fully known as to why coagonism is required for the opening of NMDAR ion channels . We show herein that the ligand binding domains ( LBD ) of the Q05586 and Q12879 subunits of the NMDAR heterodimerize only when both coagonists , DB00142 and DB00145 /d- DB00133 , bind to their respective sites on GluN2 and Q05586 . In the agonist-free state , these domains form homomeric interactions , which are disrupted by binding of their respective agonists . Also , in a heteromer formed by the LBDs , Q12879 is more sensitized to bind DB00142 , while the affinity of DB00145 for Q05586 remains unchanged . We thus provide direct evidence to show that coagonism is necessary for heteromeric pairing of LBDs , which is an essential step in forming functional ion channels in NMDARs . Localization of the succinate receptor in the distal nephron and its signaling in polarized MDCK cells . When the succinate receptor ( Q9BXA5 ) is activated in the afferent arterioles of the glomerulus it increases renin release and induces hypertension . To study its location in other nephron segments and its role in kidney function , we performed immunohistochemical analysis and found that Q9BXA5 is located in the luminal membrane of macula densa cells of the juxtaglomerular apparatus in close proximity to renin-producing granular cells , the cortical thick ascending limb , and cortical and inner medullary collecting duct cells . In order to study its signaling , Q9BXA5 was stably expressed in Madin-Darby Canine Kidney ( MDCK ) cells , where it localized to the apical membrane . Activation of the cells by succinate caused Gq and Gi-mediated intracellular calcium mobilization , transient phosphorylation of extracellular regulated kinase ( P29323 )1/2 and the release of arachidonic acid along with prostaglandins E2 and I2 . Signaling was desensitized without receptor internalization but rapidly resensitized upon succinate removal . Immunohistochemical evidence of phosphorylated P27361 /2 was found in cortical collecting duct cells of wild type but not Q9BXA5 knockout streptozotocin-induced diabetic mice , indicating in vivo relevance . Since urinary succinate concentrations in health and disease are in the activation range of the Q9BXA5 , this receptor can sense succinate in the luminal fluid . Our study suggests that changes in the luminal succinate concentration may regulate several aspects of renal function . 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 . Suppressive effect of insulin infusion on chemokines and chemokine receptors . OBJECTIVE : In view of the previously described anti-inflammatory effects of insulin , we investigated the potential suppressive effect of insulin on plasma concentrations and expression of the chemokines , monocyte chemoattractant protein-1 ( P13500 ) and regulated on activation normal T-cell expressed and secreted ( RANTES ) and their receptors , chemokine receptor ( CCR ) -2 and P51681 , in mononuclear cells ( MNCs ) . We also investigated the effect of insulin on other chemokines . RESEARCH DESIGN AND METHODS : Ten obese type 2 diabetic patients were infused with insulin ( 2 units/h with 100 ml of 5 % dextrose/h ) for 4 h . Another 8 and 6 type 2 diabetic patients were infused with 100 ml of 5 % dextrose/h or saline for 4 h , respectively , and served as control subjects . Blood samples were obtained at 0 , 2 , 4 , and 6 h . RESULTS : P01308 infusion significantly suppressed the plasma concentrations of P13500 , eotaxin , and RANTES and the expression of RANTES , macrophage inflammatory protein ( MIP ) -1beta , P41597 , and P51681 in MNCs at 2 and 4 h . DB09341 and saline infusions did not alter these indexes . CONCLUSIONS : A low-dose infusion of insulin suppresses the plasma concentration of key chemokines , P13500 , and RANTES , and the expression of their respective receptors , P41597 and P51681 , in MNCs . P01308 also suppresses the expression of RANTES and MIP-1beta in MNCs . These actions probably contribute to the comprehensive anti-inflammatory effect of insulin . Discovery of DB05130 , a Potent , Selective , and Orally Bioavailable hCCR2 Antagonist . We report the identification of 13 ( DB05130 ) as a potent human P41597 ( hCCR2 ) antagonist . DB05130 exhibited an IC50 of 3.7 nM in antagonism of monocyte chemoattractant protein-1 binding to hCCR2 , an IC50 of 4.7 nM in antagonism of chemotaxis activity , an IC50 of 84 μM in inhibition of the hERG potassium current , a free fraction of 58 % in protein binding , high selectivity over other chemokine receptors and G-protein-coupled receptors , and acceptable oral bioavailability in rodents and primates . In human clinical trials , DB05130 exhibited a pharmacokinetic profile suitable for once-a-day dosing ( T 1/2 = 15 h ) . Inhibition of rat renal fibroblast proliferation by halofuginone . BACKGROUND/AIM : Interstitial fibrosis is the final common pathway of renal damage and represents an important therapeutic target . DB04866 is a nontoxic alkaloid , used as a coccidiostat , and is a potent inhibitor of collagen alpha(1)(I) and matrix metalloproteinase-2 ( P08253 ) expression . We thus studied the effects of halofuginone on proliferation , collagen I synthesis , and P08253 activity of rat renal papillary fibroblasts in culture . METHODS : Fibroblasts were isolated from rat renal papillae and studied during passages 3-4 . The cell proliferation was studied in the presence of varying concentrations of halofuginone . The collagen synthesis was studied by [3H]proline uptake , before and after collagenase digestion , at varying concentrations of halofuginone . The P08253 activity was determined by zymography . The gelatinolytic activity was determined on gelatin-impregnated polyacrylamide gels containing samples of cell medium after incubation for 24 h with different halofuginone doses . RESULTS : We studied a phenotype of papillary fibroblasts which stained positive for alpha smooth muscle actin . These cells are phenotypically myofibroblasts . Halufuginone inhibited the proliferation of these cells in a dose-related and reversible manner . Platelet-derived growth factor is known to stimulate fibroblast proliferation . DB04866 at a concentration of 250 ng/ml almost completely abolished the effect of platelet-derived growth factor . It also almost completely inhibited the P08253 activity at doses of 250-350 ng/ml , as shown by zymography . CONCLUSIONS : DB04866 exhibits antifibrotic effects in rat renal papillary fibroblasts in culture , in terms of inhibition of proliferation and inhibition of P08253 . These findings could have therapeutic potential .

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interacts_with DB00623?

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HIV induces both a down-regulation of Q9NWZ3 that impairs TLR signalling and an up-regulation of the antibiotic peptide dermcidin in monocytic cells . HIV-infected individuals have an increased risk of invasive bacterial infections , even at early clinical stages with relatively normal P01730 (+) T-cell counts . The pathogenic mechanisms behind this are not fully understood . However , an increasing number of studies indicate that HIV may impair the innate immunity to bacteria by infecting key cells of the monocyte/macrophage lineage . In this study , the effects of HIV infection on the protein profile of undifferentiated monocyte-like THP-1 cells were examined by a mass spectrometric approach based on stable isotope labelling with amino acid in cell culture ( SILAC ) . We identified 651 proteins , of which nine proteins were down-regulated and 17 proteins were up-regulated in HIV-infected THP-1 cells as compared to uninfected controls . Most remarkably , the IL-1 receptor associated kinase 4 ( Q9NWZ3 ) , which is essential for virtually all TLR signalling , was suppressed , whereas the precursor for the antibiotic peptide P81605 was up-regulated in HIV-infected cells . Upon stimulation of either O60603 or O00206 , the HIV-infected THP-1 cells displayed reduced P01375 secretion . The HIV-induced down-regulation of Q9NWZ3 was reconfirmed in monocyte-derived macrophage cell cultures . These data suggests that HIV may impair the TLR signalling cascade for pathogen recognition in cells of the monocyte/macrophage lineage and thus , may reduce the ability of the innate immune system to sense invading pathogens and initiate appropriate responses . Transplanted iNSCs migrate through P48061 / P61073 signaling to promote neural recovery in a rat model of spinal cord injury . The ability of transplanted induced neural stem cells ( iNSCs ) to promote functional recovery after spinal cord injury and the mechanism by which iNSCs migrate to injured areas are poorly understood . Stromal cell-derived factor-1 ( P48061 ) is a cytokine , whereas P61073 is its cognate receptor . The aim of this study was to determine whether P48061 regulates the migration of iNSCs and to explore the potential mechanism by which iNSCs promotes functional recovery . In-vitro experiments demonstrated that P48061 induces a concentration-dependent migration of iNSCs . Pretreatment with the P61073 -specific antagonist DB06809 significantly prevented the migration of iNSCs . We found that the expression of P48061 increased significantly in spinal cord lesions and was mainly associated with neurons and astrocytes . We also demonstrated that transplanted green fluorescent protein-labeled iNSCs were localized to regions where P48061 was highly expressed . In addition , iNSC-treated animals showed significantly improved functional recovery as assessed by BBB at 7 days after injection compared with controls . iNSCs also increased cell proliferation , enhanced vascularity , and reduced apoptosis . These results suggest that upregulated P48061 plays an important role in the migration of iNSCs to the injured region and that iNSCs are beneficial for functional recovery after spinal cord injury . Technique appraisement of comparative proteomics and screening of differentiation-related protein in gastric carcinoma . BACKGROUND/AIMS : Different differentiations of cancer have resulted in its unique biological characteristics . We screen and appraise differentially expressed proteins in different differentiated gastric adenocarcinoma with comparative proteomics technology in order to find regulatory factors of tumor differentiation related and finally reach the purpose of tumor differentiation reversal . METHODOLOGY : With two-dimensional fluorescence difference gel electrophoresis ( 2-D DIGE ) and liquid chromatography in conjunction with tandem mass spectrometry ( LC–MS/MS ) , the differentially expressed proteins from 8 patients with different differentiated gastric adenocarcinoma were identified and some factors identified were verified with application of QPCR and Western blot techniques . RESULTS : Significant differences in 35 protein spots were found and 48 kinds of proteins were identified . Other than structural proteins and non-specific protein , six possible proteins associated with tumor differentiation were determined - the serine protease inhibitor B1 ( serine protease inhibitor , clade B , member 1 , P30740 ) , calcium-phospholipid binding protein III ( annexin A3 ) , transcription factor Nm23-H1 , adenine phosphoribosyl-transferase enzyme P07741 ( DB00173 Phosphoribosyltransferase in APO and AMP ) , glutathione S-transferase P1-1 ( Q86UG4 -π-1 ) , antimicrobial peptides P81605 -lL . The identified P30740 , annexin A3 , Nm23-H1 and P07741 were verified , confirming the expression of these factors was in line with proteomics identification . CONCLUSIONS : Protein expression in different differentiated gastric adenocarcinoma was varied . Functional characterization of methionine sulfoxide reductase A from Trypanosoma spp . DB00134 is an amino acid susceptible to being oxidized to methionine sulfoxide ( MetSO ) . The reduction of MetSO to methionine is catalyzed by methionine sulfoxide reductase ( Q9UBK8 ) , an enzyme present in almost all organisms . In trypanosomatids , the study of antioxidant systems has been mainly focused on the involvement of trypanothione , a specific redox component in these organisms . However , no information is available concerning their mechanisms for repairing oxidized proteins , which would be relevant for the survival of these pathogens in the various stages of their life cycle . We report the molecular cloning of three genes encoding a putative A-type Q9UBK8 in trypanosomatids . The genes were expressed in Escherichia coli , and the corresponding recombinant proteins were purified and functionally characterized . The enzymes were specific for L- DB00134 (S)SO reduction , using Trypanosoma cruzi tryparedoxin I as the reducing substrate . Each enzyme migrated in electrophoresis with a particular profile reflecting the differences they exhibit in superficial charge . The in vivo presence of the enzymes was evidenced by immunological detection in replicative stages of T. cruzi and Trypanosoma brucei . The results support the occurrence of a metabolic pathway in Trypanosoma spp. involved in the critical function of repairing oxidized macromolecules . Leukotriene D4 upregulates Q02817 gene transcription in human epithelial cells . BACKGROUND/AIMS : Leukotriene ( LT ) D(4) has been shown to induce mucus secretion in the airways . Excessive mucus secretion characterizes airway inflammatory disease such as asthma , allergic rhinitis . However , little is known about the effect of LTD(4) on mucin gene expression . The aim of this study was to investigate the effect of LTD(4) on Q02817 gene expression in cultured epithelial cells ( HM3- Q02817 cells ) . METHODS : HM3- Q02817 cells were treated with LTD(4) for 2 or 6 h . Reporter gene assay was mainly used for analysis. Q02817 protein levels were measured using an enzyme-linked immunosorbent assay . RESULTS : LTD(4) significantly increased Q02817 gene transcriptional activity in a dose-dependent manner . DB01411 , which is a selective antagonist of CysLT(1) receptor , inhibited LTD(4)-induced Q02817 gene transcriptional activity in a dose-dependent manner . LTD(4)-induced Q02817 gene transcriptional activity was also suppressed by a G-protein inhibitor ( pertussis toxin ) ,a protein kinase C ( PKC ) inhibitor ( bisindolylmaleimide ) , a mitogen-activated protein/extracellular signal-regulated kinase kinase ( MEK ) inhibitor ( PD98059 ) , an extracellular signal regulated kinase-2 ( P28482 ) inhibitor ( AG126 ) and a nuclear factor kappaB ( NF-kappaB ) inhibitor . In addition , pranlukast inhibited LTD(4)-induced NF-kappaB activity . CONCLUSION : These results suggest that LTD ( 4 ) upregulates Q02817 gene transcription via a signaling pathway involving CysLT(1) receptor , G-protein , PKC , MEK , P29323 and NF-kappaB . 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 . Hypothalamic expression of serotonin 1A , 2A and 2C receptor and Q99259 mRNA in female cynomolgus monkeys with different sensitivity to stress . Like women , female cynomolgus monkeys show differential sensitivity to stress-induced reproductive dysfunction . A combined social and metabolic stress ( mild diet+moderate exercise+relocation ) will rapidly induce anovulation in a third of female cynomolgus monkeys ( stress-sensitive ; SS ) ; a third will ovulate once and then become anovulatory ( medium stress-resilient ; Q9UBK8 ) and a third are highly stress-resilient ( HSR ) and exhibit normal menstrual cycles through two stressed menstrual cycles . In a non-stressed menstrual cycle , SS animals have lower levels of estrogen and progesterone , lower activity of the serotonin system and lower expression of genes related to the serotonin system in the dorsal raphe nucleus . In this study , we examined the expression of 5HT1A , 5HT2A , 5HT2C receptors and Q99259 in the hypothalamus of SS , HSR and Q9UBK8 monkeys using in situ hybridization . SS monkeys exhibited higher expression of 5HT2A mRNA in the paraventricular nucleus ( PVN ) , higher expression of 5HT2C and Q99259 in the infundibulum , as well as higher expression of Q99259 in the posterior hypothalamus ( PH ) , compared with HSR monkeys . However , the expression of 5HT1A mRNA in the ventromedial nucleus ( VMN ) was not different between groups . We speculate that the serotonin and GABA systems may be altered in the stress-response and reproductive-related circuits of SS monkeys , and may be participating in altering the sensitivity of the reproductive system to stress in these individuals . Activation of large-conductance , Ca2+-activated K+ channels by cannabinoids . We have examined the effects of the cannabinoid anandamide ( AEA ) and its stable analog , methanandamide ( methAEA ) , on large-conductance , Ca2+-activated K+ ( BK ) channels using human embryonic kidney ( P29320 ) -293 cells , in which the alpha-subunit of the Q12791 ( BK-alpha ) , both alpha- and beta1-subunits ( BK-alphabeta1 ) , or both alpha- and beta4-subunits ( BK-alphabeta4 ) were heterologously expressed . In a whole cell voltage-clamp configuration , each cannabinoid activated BK-alphabeta1 within a similar concentration range . Because methAEA could potentiate BK-alpha , BK-alphabeta1 , and BK-alphabeta4 with similar efficacy , the beta-subunits may not be involved at the site of action for cannabinoids . Under cell-attached patch-clamp conditions , application of methAEA to the bathing solution increased Q12791 activity ; however , methAEA did not alter channel activity in the excised inside-out patch mode even when DB00171 was present on the cytoplasmic side of the membrane . Application of methAEA to P29320 -BK-alpha and P29320 -BK-alphabeta1 did not change intracellular Ca2+ concentration . Moreover , methAEA-induced potentiation of Q12791 currents was not affected by pretreatment with a P21554 antagonist ( AM251 ) , modulators of G proteins ( cholera and pertussis toxins ) or by application of a selective CB2 agonist ( JWH133 ) . Inhibitors of P62158 , PKG , and MAPKs ( W7 , KT5823 , and PD-98059 ) did not affect the potentiation . Application of methAEA to mouse aortic myocytes significantly increased Q12791 currents . This study provides the first direct evidence that unknown factors in the cytoplasm mediate the ability of endogenous cannabinoids to activate Q12791 currents . Cannabinoids may be hyperpolarizing factors in cells , such as arterial myocytes , in which BK channels are highly expressed . Ghrelin protects heart against ERS-induced injury and apoptosis by activating AMP-activated protein kinase . Ghrelin , the endogenous ligand of growth hormone secretagogue receptor ( Q92847 ) , is a cardioprotective peptide . In our previous work , we have revealed that ghrelin could protect heart against ischemia/reperfusion ( I/R ) injury by inhibiting endoplasmic reticulum stress ( ERS ) , which contributes to many heart diseases . In current study , using both in vivo and in vitro models , we investigated how ghrelin inhibits myocardial ERS . In the in vivo rat heart injury model induced by isoproterenol ( ISO ) , we found that exogenous ghrelin could alleviate heart dysfunction , reduce myocardial injury and apoptosis and inhibit the excessive myocardial ERS induced by ISO . More importantly , the activation of AMP-activated protein kinase ( AMPK ) was observed . To explore the role of AMPK activation in ERS inhibition by ghrelin , we set up two in vitro ERS models by exposing cultured rat cardiomyocytes to tunicamycin(Tm) or dithiothreitol ( DTT ) . In both models , compared with Tm or DTT treatment alone , pre-incubation cardiomyocytes with ghrelin significantly activated AMPK , reversed the upregulation of the ERS markers , P35638 ( P35638 ) and cleaved caspase-12 , and reduced apoptosis of cardiomyocytes . Further , we found that the ERS inhibitory and anti-apoptotic actions induced by ghrelin were blocked by an AMPK inhibitor . To investigate how ghrelin activates AMPK , selective antagonist of GHS-R1a and inhibitor of Ca(2+)/ P62158 -dependent protein kinase kinase ( CaMKK ) were added , respectively , before ghrelin pre-incubation , and we found that AMPK activation was prevented and the ERS inhibitory and anti-apoptotic actions of ghrelin were blocked . In conclusion , ghrelin could protect heart against ERS-induced injury and apoptosis , at least partially through a GHS-R1a/CaMKK/AMPK pathway . [ Response to antitumor agents of human transplantable glioma implanted into chorioallantoic membrane of chick embryo ] . In case of chemotherapy against brain tumors , it is most important to choose suitable drugs for brain tumors , since human tumors have different drug sensitivity and growth . Heretofore , the nitrosourea-induced rat glioma cell , such as P13671 , or immunodeficient mice were usually used for predicting the drug sensitivity of brain tumors . We took notice of Murphy 's system for the chemosensitivity test , in which a human tumor is transplanted into the chorioallantoic membrane ( P62158 ) of a chick embryo . By modifying the conventional Murphy 's system , we studied the efficiency of this system in predicting the drug sensitivity of brain tumors . First , we compared the result of a drug sensitivity test using P62158 of a chick embryo with that using nude mice . Next we studied the effect of chemotherapeutic agents against brain metastasis of a chick embryo caused by the intravenous injection of mouse B16 melanoma cells . The tumor reduction rate of the sensitivity test using a chick embryo tended to agree with that using nude mice . In the drug sensitivity test against brain metastasis , ACNU was the most effective . This result supports the result of the clinical study . In conclusion , the drug sensitivity test using a chick embryo is thought to be useful and the advantages or disadvantages of this system are discussed . 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 . 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 . 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 . P62158 interacts with the cytoplasmic tails of the parathyroid hormone 1 receptor and a sub-set of class b G-protein coupled receptors . Parathyroid hormone ( PTH ) binds to its receptor ( PTH 1 receptor , Q03431 ) and activates multiple pathways . The Q03431 , a class b GPCR , contains consensus calmodulin-binding motifs . The Q03431 cytoplasmic tail interacts with calmodulin in a calcium-dependent manner via the basic 1-5-8-14 motif . DB01373 -dependent calmodulin interactions with the cytoplasmic tails of receptors for Q9Y3E5 , vasoactive intestinal peptide , pituitary adenylate cyclase activating peptide , corticotropin releasing hormone , calcitonin , and the glucagon-like peptides 1 and 2 are demonstrated . The cytoplasmic tails of the secretin receptor and the growth hormone releasing hormone receptor either interact poorly or not at all with calmodulin , respectively . DB00623 , a calmodulin antagonist , enhances PTH-mediated accumulation of total inositol phosphates , suggesting that calmodulin regulates signaling via phospholipase C . 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 . Failure of anti-interleukin-1 therapy in severe hidradenitis suppurativa : a case report . Hidradenitis suppurativa ( HS ) is an inflammatory , debilitating skin disease of follicular origin , characterized by painful deep-seated , inflamed lesions in mainly the inverse areas of the body . HS is notoriously difficult to treat and especially severe disease is often resistant to therapy . New therapeutic options are therefore highly needed . Elevated levels of IL-1 have been demonstrated in HS lesions . Here we report for the first time on the sequential treatment with anakinra ( an IL-1 receptor antagonist ) and DB06674 ( a P01375 -α-neutralizing antibody ) of a patient with severe HS and with comorbid psoriatic arthritis . Although adalimumab and DB06674 were efficacious in improving arthritis complaints , both failed in improving the severe HS of our patient . Eventually the patient underwent radical excision of the inflammatory lesions and fistulas . 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 . 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 . 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 . 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 )

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P35354 polymorphisms , aspirin treatment , and risk for colorectal adenoma recurrence -- data from a randomized clinical trial . P35354 ( P35354 ) catalyzes the rate-limiting step in the production of prostaglandins , potent mediators of inflammation . Chronic inflammation plays an important role in the development and progression of colorectal cancer . DB00945 inhibits P35354 activity and lowers the risk for colorectal adenomas and cancer . We investigated whether common genetic variation in P35354 influenced risk for colorectal adenoma recurrence among 979 participants in the DB00945 / DB00158 Polyp Prevention Study who were randomly assigned to placebo or aspirin and followed for 3 years for the occurrence of new adenomas . Of these participants , 44.2 % developed at least one new adenoma during follow-up . Adjusted relative risks and 95 % confidence intervals ( 95 % CI ) were calculated to test the association between genetic variation at six P35354 single-nucleotide polymorphisms and adenoma occurrence and interaction with aspirin treatment . Two single-nucleotide polymorphisms were significantly associated with increased adenoma recurrence : for rs5277 , homozygous carriers of the minor C allele had a 51 % increased risk compared with GG homozygotes ( relative risk , 1.51 ; 95 % CI , 1.01-2.25 ) , and for rs4648310 , heterozygous carriers of the minor G allele had a 37 % increased risk compared with AA homozygotes ( relative risk , 1.37 ; 95 % CI , 1.05-1.79 ) . ( There were no minor allele homozygotes. ) In stratified analyses , there was suggestive evidence that rs4648319 modified the effect of aspirin . These results support the hypothesis that P35354 plays a role in the etiology of colon cancer and may be a target for aspirin chemoprevention and warrant further investigation in other colorectal adenoma and cancer populations . 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 . 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 . 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 . DNA methylation and Yin Yang-1 repress adenosine A2A receptor levels in human brain . DB00640 A(2A) receptors ( A(2A) Rs ) are G-protein coupled receptors that stimulate adenylyl cyclase activity . The most A(2A) Rs-enriched brain region is the striatum , in which A(2A) Rs are largely restricted to GABAergic neurons of the indirect pathway . We recently described how DNA methylation controls basal A(2A) R expression levels in human cell lines . The present report provides clues about the molecular mechanisms that promote human brain region-specific A(2A) R gene ( P29274 ) basal expression . The transcription factors ZBP-89 and Yin Yang-1 ( P25490 ) have been characterized as regulators of P29274 in SH-SY5Y cells by means of specific expression vectors/siRNAs transient transfection and chromatin immunoprecipitation assay . ZBP-89 plays a role as an activator and P25490 as a repressor . No differences were found in ZBP-89 levels with western blot between the putamen and cerebellum of human postmortem brains . However , increased P25490 levels and DNA methylation percentage in the 5' untranslated region of P29274 , using SEQUENOM MassArray , were found in the cerebellum with respect to the putamen of human brains , showing an inverse relationship with A(2A) R levels in the two cerebral regions . Role of human aquaporin 5 in colorectal carcinogenesis . While overexpression of several aquaporins ( AQPs ) has been reported in different types of human cancer , the role of AQPs in carcinogenesis has not been clearly defined . Here , by immunochemistry , we have found expression of P55064 protein in 62.8 % ( 59/94 ) of resected colon cancer tissue samples as well as association of P55064 with liver metastasis . We then demonstrated that overexpression of human P55064 ( hAQP5 ) induces cell proliferation in colon cancer cells . Overexpression of wild-type hAQP5 increased proliferation and phosphorylation of extracellular signal-regulated kinase-1/2 in HCT116 colon cancer cells whereas these phenomena in hAQP5 mutants ( N185D and S156A ) were diminished , indicating that both membrane association and serine/threonine phosphorylation of P55064 are required for proper function . Interestingly , overexpression of P29972 and Q92482 showed no differences in extracellular signal-regulated kinase-1/2 phosphorylation , suggesting that P55064 , unlike P29972 , may be involved in signal transduction . Moreover , hAQP5-overexpressing cells showed an increase in retinoblastoma protein phosphorylation through the formation of a nuclear complex with cyclin D1 and P11802 . Small interfering RNA analysis confirmed that hAQP5 activates the Ras signaling pathway . These data not only describe the induction of hAQP5 expression during colorectal carcinogenesis but also provide a molecular mechanism for colon cancer development through the interaction of hAQP5 with the Ras/extracellular signal-regulated kinase/retinoblastoma protein signaling pathway , identifying hAQP5 as a novel therapeutic target . 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 . 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 . [ 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 . Erythropoietin induces the tyrosine phosphorylation of Q13480 and its association with P29353 , SHP2 , Q92835 , and phosphatidylinositol 3-kinase . Five tyrosine-phosphorylated proteins with molecular masses of 180 , 145 , 116 , 100 , and 70 kD are associated with phosphatidylinositol 3-kinase ( PI 3-kinase ) in erythropoietin ( Epo ) -stimulated UT-7 cells . The 180- and 70-kD proteins have been previously shown to be Q9Y4H2 and the Epo receptor . In this report , we show that the 116-kD protein is the Q9Y4H2 -related molecular adapter , Q13480 . Indeed , Epo induced the transient tyrosine phosphorylation of Q13480 in UT-7 cells . Both kinetics and Epo dose-response experiments showed that Q13480 tyrosine phosphorylation was a direct consequence of Epo receptor activation . After tyrosine phosphorylation , Q13480 associated with the PI 3-kinase , the phosphotyrosine phosphatase SHP2 , the phosphatidylinositol 3,4,5 trisphosphate 5-phosphatase Q92835 , and the molecular adapter P29353 . Q13480 was also associated with the molecular adapter P62993 in unstimulated cells , and this association dramatically increased after Epo stimulation . Thus , Q13480 could be a scaffold protein able to couple the Epo receptor activation with the stimulation of several intracellular signaling pathways . Epo-induced tyrosine phosphorylation of Q13480 was also observed in normal human erythroid progenitors isolated from cord blood . P04141 ( GM- P04141 ) and thrombopoietin ( P07202 ) also induced the tyrosine phosphorylation of Q13480 in UT-7 cells , indicating that this molecule participates in the signal transduction of several cytokine receptors . Phase II study of weekly intravenous oxaliplatin combined with oral daily capecitabine and radiotherapy with biologic correlates in neoadjuvant treatment of rectal adenocarcinoma . PURPOSE : To evaluate the efficacy of a combination of capecitabine , oxaliplatin , and radiotherapy ( RT ) in the neoadjuvant treatment of Stage II and III rectal cancers . METHODS : DB01101 was given at 725 mg/m(2) orally twice daily Monday through Friday concurrently with RT . DB00526 was given intravenously at 50 mg/m(2) once weekly five times starting the first day of RT . The radiation dose was 50.4 Gy in 28 fractions ( 1.8 Gy/fraction ) , five fractions weekly . Endorectal tumor biopsies were obtained before treatment and on the third day of treatment to explore the effects of treatment on thymidine phosphorylase , thymidylate synthase , excision repair cross-complementing rodent repair deficiency complementation group 1 ( P07992 ) , and apoptosis . RESULTS : A total of 25 patients were enrolled in this study ; 6 patients ( 24 % ) had a complete pathologic response . T-downstaging occurred in 52 % of patients , and N-downstaging occurred in 53 % . Grade 3 diarrhea was the most common Grade 3-4 toxicity , occurring in 20 % of patients . Only 2 patients experienced disease recurrence , with a median of 20 months of follow-up . P04818 , thymidine phosphorylase , P07992 , and apoptosis did not vary significantly between the pretreatment and Day 3 tumor biopsies , nor did they predict for T-downstaging or a complete pathologic response . CONCLUSION : DB01101 at 725 mg/m(2) orally twice daily , oxaliplatin 50 mg/m(2)/wk , and RT at 50.4 Gy is an effective neoadjuvant combination for Stage II and III rectal cancer and results in a greater rate of complete pathologic responses than historically shown in fluoropyrimidine plus RT controls . DB00203 protects human mammary epithelial cells against ROS production induced by estradiol . Several studies suggest that xanthine dehydrogenase ( P47989 ) and its oxidase form ( XO ) play an important role in various types of ischemic and vascular injuries . Recently , we have demonstrated that estradiol ( E2 ) induces a significant decrease of the expression and activity of P47989 and of its conversion to XO in human mammary epithelial cells . E2 is known to induce upregulation of P29474 gene expression in aortic endothelial cells . Because the XO-derived O2·- combines with ·NO to yield ONOO- , and considering that ONOO- converts P47989 to XO , the resulting increase of XO activity and reactive oxygen species production would eventually lead to a further increase of ONOO- production , thus creating a vicious cycle of oxidative stress . Our previous study has indicated that sildenafil has a protective effect on human mammary epithelial cells as a consequence of XO inhibition and of the resulting decrease of free oxygen radicals that can impair the expression of NADPH oxidase and type 5 phosphodiesterase ( PDE-5 ) . In the present study , we report that the dual inhibitory effect exerted by sildenafil on both XO and PDE-5 is a consequence of a structural modification induced by O2·- , also consisting of the release of a piperazine group that could in turn inhibit the XO enzyme . P55008 arrest and down-regulation of cyclin E/cyclin-dependent kinase 2 by the protein kinase inhibitor staurosporine are dependent on the retinoblastoma protein in the bladder carcinoma cell line 5637 . The protein kinase inhibitor staurosporine has been shown to induce P55008 phase arrest in normal cells but not in most transformed cells . DB02010 did not induce P55008 phase arrest in the bladder carcinoma cell line 5637 that lacks a functional retinoblastoma protein ( pRB- ) . However , when infected with a pRB-expressing retrovirus [ Goodrich , D. W. , Chen , Y. , Scully , P. & Lee , W.-H. ( 1992 ) Cancer Res. 52 , 1968-1973 ] , these cells , now pRB+ , were arrested by staurosporine in P55008 phase . This arrest was accompanied by the accumulation of hypophosphorylated pRB . In both the pRB+ and pRB- cells , cyclin D1-associated kinase activities were reduced on staurosporine treatment . In contrast , cyclin-dependent kinase ( CDK ) 2 and cyclin E/ P24941 activities were inhibited only in pRB+ cells . DB02010 treatment did not cause reductions in the protein levels of P11802 , cyclin D1 , P24941 , or cyclin E . The CDK inhibitor proteins P38936 (Waf1/Cip1) and p27 ( Kip1 ) levels increased in staurosporine-treated cells . Immunoprecipitation of P24941 , cyclin E , and p2l from staurosporine-treated pRB+ cells revealed a 2.5- to 3-fold higher ratio of p2l bound to P24941 compared with staurosporine-treated pRB- cells . In pRB+ cells , p2l was preferentially associated with Thrl6O phosphorylated active P24941 . In pRB- cells , however , p2l was bound preferentially to the unphosphorylated , inactive form of P24941 even though the phosphorylated form was abundant . This is the first evidence suggesting that P55008 arrest by 4 nM staurosporine is dependent on a functional pRB protein . Cell cycle arrest at the pRB- dependent checkpoint may prevent activation of cyclin E/ P24941 by stabilizing its interaction with inhibitor proteins p2l and p27 . Quantum dots trigger immunomodulation of the NFκB pathway in human skin cells . The immunological effects of quantum dots are dependent on a variety of factors including , but not limited to , exposure time and dosing concentrations . In this study , we investigated the influence of 15 nm CdSe/ZnS-COOH quantum dot nanocrystals ( QDs ) on cell density , viability , and morphology in human epidermal keratinocytes ( P29320 ) and human dermal fibroblasts ( HDF ) . Furthermore , inflammatory and non-inflammatory immune responses were measured using protein and real time PCR array analysis from HDF cells exposed to predetermined sub-lethal concentrations of QDs . CdSe/ZnS-COOH QDs caused concentration-dependent ( 1-120 nM exposure concentrations ) and time-dependent ( 8 h or 48 h ) cell death , as evidenced by metabolic activity and morphological changes . QD exposure induced upregulation of apoptotic , inflammatory and immunoregulatory proteins such as P01375 -α , IL-1B and P22301 . P09601 , an indicator of stress due to reactive oxygen intermediates ( ROIs ) and/or metals , was upregulated at the later time point as well . QDs also caused modulation of genes known to be associated with inflammatory ( IL1-β , P13500 , O43187 ) , immune ( IL-1 , P05231 , O75594 , P01009 , P22301 ) , stress due to ROIs and/or heavy metals ( P09601 ) , and apoptotic ( P29466 , P29274 ) responses . Cellular effects from QD exposure were found to primarily follow the NFκB pathway . In addition , QDs induced a differential cytotoxicity in keratinocytes and fibroblasts at different exposure concentrations and time points , even at physiologically relevant dosing concentrations , thus emphasizing the need to investigate potential mechanisms of action among different cell types within the same target organ . Distinct effects of inflammation on gliosis , osmohomeostasis , and vascular integrity during amyloid beta-induced retinal degeneration . In normal retinas , amyloid-β ( Aβ ) accumulates in the subretinal space , at the interface of the retinal pigment epithelium , and the photoreceptor outer segments . However , the molecular and cellular effects of subretinal Aβ remain inadequately elucidated . We previously showed that subretinal injection of Aβ(1-42) induces retinal inflammation , followed by photoreceptor cell death . The retinal Müller glial ( RMG ) cells , which are the principal retinal glial cells , are metabolically coupled to photoreceptors . Their role in the maintenance of retinal water/potassium and glutamate homeostasis makes them important players in photoreceptor survival . This study investigated the effects of subretinal Aβ(1-42) on RMG cells and of Aβ(1-42)-induced inflammation on retinal homeostasis . RMG cell gliosis ( upregulation of P14136 , vimentin , and nestin ) on day 1 postinjection and a proinflammatory phenotype were the first signs of retinal alteration induced by Aβ(1-42) . On day 3 , we detected modifications in the protein expression patterns of cyclooxygenase 2 ( P35354 ) , glutamine synthetase ( GS ) , Kir4.1 [ the inwardly rectifying potassium ( Kir ) channel ] , and aquaporin ( AQP ) -4 water channels in RMG cells and of the photoreceptor-associated P29972 . The integrity of the blood-retina barrier was compromised and retinal edema developed . Aβ(1-42) induced endoplasmic reticulum stress associated with sustained upregulation of the proapoptotic factors of the unfolded protein response and persistent photoreceptor apoptosis . Indomethacin treatment decreased inflammation and reversed the Aβ(1-42)-induced gliosis and modifications in the expression patterns of P35354 , Kir4.1 , and P29972 , but not of P55087 or GS . Nor did it improve edema . Our study pinpoints the adaptive response to Aβ of specific RMG cell functions . Retinoid receptors and acute promyelocytic leukaemia . While a great deal has been learned about APL over the last few years , many important questions remain unanswered . It has become clear that the P29590 / P10276 fusion protein is expressed in most cases of APL , and this protein presumably contributes to leukaemia initiation and/or progression . P29590 / P10276 appears to specifically block the further differentiation of myeloid progenitor cells , although the mechanism of its action is not known . It may inhibit the transcription of RAR- or P29590 -regulated genes , in which case expression must be restored in the presence of therapeutic RA concentrations . However , the possibility remains that P29590 / P10276 may have a novel function . In order to elucidate the molecular pathogenesis of APL , several important questions remain to be answered . These include whether P29590 is a transcription factor ; the identification of its target genes and response elements , and the role of P29590 / P10276 and RA in their regulation . Also whether the expression of P29590 / P10276 in bone marrow cells ( either by itself or in combination with other oncogenes ) alters their tumourigenicity or differentiation potential . It is also important to determine the function(s) of Q05516 and Q05516 / P10276 , and determine whether other APL patients with mutations involving P29590 or P10276 ( but not both ) respond to therapy with all-trans-RA . Finally , it is important both for the understanding of the molecular biology of APL and its therapy , to determine the effects of other regulatory factors involved in the control of myeloid cell differentiation such as granulocyte colony stimulation factor ( DB00099 ) and granulocyte macrophage colony stimulating factor ( GM- P04141 ) on APL cells in vitro and in vivo , both at presentation and in the RA-resistant patients in relapse . P29972 in the peritoneal membrane : implications for peritoneal dialysis and endothelial cell function . PD ( peritoneal dialysis ) is an established mode of renal replacement therapy , based on the exchange of fluid and solutes between blood in peritoneal capillaries and a dialysate that has been introduced into the peritoneal cavity . The dialysis process involves diffusive and convective transports and osmosis through the PM ( peritoneal membrane ) . Computer simulations predicted that the PM contains ultrasmall pores ( radius < 3 A , 1 A=10(-10) m ) , responsible for up to 50 % of UF ( ultrafiltration ) , i.e. the osmotically driven water movement during PD . Several lines of evidence suggest that P29972 ( aquaporin-1 ) is the ultrasmall pore responsible for transcellular water permeability during PD . Treatment with corticosteroids induces the expression of P29972 in the PM and improves water permeability and UF in rats without affecting the osmotic gradient and permeability for small solutes . Studies in knockout mice provided further evidence that osmotically driven water transport across the PM is mediated by P29972 . P29972 and P29474 ( endothelial nitric oxide synthase ) show a distinct regulation within the endothelium lining the peritoneal capillaries . In acute peritonitis , the up-regulation of P29474 and increased release of nitric oxide dissipate the osmotic gradient and prevent UF , whereas P29972 expression is unchanged . These results illustrate the usefulness of the PM to investigate the role and regulation of P29972 in the endothelium . The results also emphasize the critical role of P29972 during PD and suggest that manipulation of P29972 expression may be used to increase water permeability across the PM . The effects of aging and chronic fluoxetine treatment on circadian rhythms and suprachiasmatic nucleus expression of neuropeptide genes and P28222 receptors . Age-related changes in circadian rhythms , including attenuation of photic phase shifts , are associated with changes in the central pacemaker in the suprachiasmatic nucleus ( SCN ) . Aging decreases expression of mRNA for vasoactive intestinal peptide ( P01282 ) , a key neuropeptide for rhythm generation and photic phase shifts , and increases expression of serotonin transporters and 5-HT(1B) receptors , whose activation inhibits these phase shifts . Here we describe studies in hamsters showing that aging decreases SCN expression of mRNA for gastrin-releasing peptide , which also modulates photic phase resetting . Because serotonin innervation trophically supports SCN P01282 mRNA expression , and serotonin transporters decrease extracellular serotonin , we predicted that chronic administration of the serotonin-selective reuptake inhibitor , fluoxetine , would attenuate the age-related changes in SCN P01282 mRNA expression and 5-HT(1B) receptors . In situ hybridization studies showed that fluoxetine treatment does not alter SCN P01282 mRNA expression , in either age group , at zeitgeber time (ZT)6 or 13 ( ZT12 corresponds to lights off ) . However , receptor autoradiographic studies showed that fluoxetine prevents the age-related increase in SCN 5-HT(1B) receptors at ZT6 , and decreases SCN 5-HT(1B) receptors in both ages at ZT13 . Therefore , aging effects on SCN P01282 mRNA and SCN 5-HT(1B) receptors are differentially regulated ; the age-related increase in serotonin transporter sites mediates the latter but not the former . The studies also showed that aging and chronic fluoxetine treatment decrease total daily wheel running without altering the phase of the circadian wheel running rhythm , in contrast to previous reports of phase resetting by acute fluoxetine treatment . Expression of Bcl-2 predicts outcome in locally advanced non-small cell lung cancer patients treated with cisplatin-based concurrent chemoradiotherapy . BACKGROUND : Platinum-based concurrent chemoradiotherapy ( CCRT ) is a standard treatment for locally advanced unresectable non-small cell lung cancer ( NSCLC ) . The determination of parameters that may predict the result of the treatment has strong clinical implications . PATIENTS AND METHODS : Pretreatment tumor biopsy specimens from 39 patients with locally advanced NSCLC ( stage IIIA : 5 , stage IIIB : 34 ) were analyzed for p53 , Bcl-2 , Bax and P07992 expression by immunohistochemistry . All patients were treated with cisplatin-based CCRT . Twenty-four patients received induction chemotherapy followed by CCRT ( 60Gy/30 fractions , 6mg/m(2) of cisplatin daily ) . The most commonly administered induction chemotherapy regimen was P01282 ( etoposide , ifosfamide , cisplatin ; 20 patients ) . Fifteen patients received the same CCRT without induction chemotherapy . RESULTS : High expression of p53 , Bcl-2 , Bax and P07992 was observed in 15 ( 38 % ) , 19 ( 49 % ) , 17 ( 44 % ) and 12 ( 31 % ) patients , respectively . High expression of Bcl-2 was significantly associated with longer survival duration ( 20 months vs. 9 months , P=0.008 ) and better response to the treatment ( 74 % vs. 30 % , P=0.01 ) . In multivariate analysis , Bcl-2 expression was the only significant independent prognostic factor of overall survival ( P=0.007 ) among the pretreatment patients characteristics . CONCLUSIONS : High expression of Bcl-2 may be a useful prognostic factor in locally advanced NSCLC patients treated with cisplatin-based CCRT . Poly(ADP-ribose) polymerase-1 inhibition prevents eosinophil recruitment by modulating Th2 cytokines in a murine model of allergic airway inflammation : a potential specific effect on P05113 . We recently used a murine model of allergic airway inflammation to show that poly(ADP-ribose) polymerase-1 ( P09874 ) plays an important role in the pathogenesis of asthma-related lung inflammation . In this study , we show that P09874 inhibition , by a novel inhibitor ( TIQ-A ) or by gene deletion , prevented eosinophilic infiltration into the airways of OVA-challenged mice . Such impairment of eosinophil recruitment appeared to take place after IgE production . OVA challenge of wild-type mice resulted in a significant increase in P05112 , P05113 , P22301 , P35225 , and GM- P04141 secretions . Although P05112 production was moderately affected in OVA-challenged P09874 (-/-) mice , the production of P05113 , P22301 , P35225 , and GM- P04141 was completely inhibited in ex vivo OVA-challenged lung cells derived from these animals . A single TIQ-A injection before OVA challenge in wild-type mice mimicked the latter effects . The marked effect P09874 inhibition exerted on mucus production corroborated the effects observed on the Th2 response . Although P09874 inhibition by gene knockout increased the production of the Th1 cytokines P60568 and IL-12 , the inhibition by TIQ-A exerted no effect on these two cytokines . The failure of lung cells derived from OVA-challenged P09874 (-/-) mice to synthesize GM- P04141 , a key cytokine in eosinophil recruitment , was reestablished by replenishment of P05113 . Furthermore , intranasal administration of P05113 restored the impairment of eosinophil recruitment and mucus production in OVA-challenged P09874 (-/-) mice . The replenishment of either P05112 or IgE , however , did not result in such phenotype reversals . Altogether , these results suggest that P09874 plays a critical role in eosinophil recruitment by specifically regulating the cascade leading to P05113 production . 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 . Differential effects of all-trans and 13-cis-retinoic acid on mRNA levels of nuclear retinoic acid receptors in rat lung and liver . The effects of three retinoids , all-trans-retinoic acid ( all-trans-RA ) , 13- DB00982 , and etretin were examined on mRNA abundance of nuclear retinoic acid receptors ( P10276 , beta , and gamma ) in lung and liver of retinol deficient and chow fed rats . All-trans-RA increased lung P10826 mRNA levels 5 or 11-fold in chow fed and retinol deficient rats , respectively . Similarly to lung , liver P10826 mRNA levels were 3-fold higher in retinol deficient rats fed all-trans-RA than the rats fed cottonseed oil . Lung P13631 mRNA levels were also induced 2-fold by all-trans-RA . In contrast to this , 13- DB00982 and etretin at equimolar doses failed to enhance lung or liver P10826 or lung P13631 mRNA levels in retinol deficient rats . These data for the first time show that all-trans-RA is more effective than its 13-cis-isomer in regulating the expression of P10826 and gamma transcripts in adult animal . Role for macrophage migration inhibitory factor in acute respiratory distress syndrome . The critical role of macrophage migration inhibitory factor ( MIF ) in mediating inflammatory lung injury in acute respiratory distress syndrome ( ARDS ) has been raised recently . The present study has identified enhanced MIF protein expression in alveolar capillary endothelium and infiltrating macrophages in lung tissues from ARDS patients . The possibility that MIF up-regulates its synthesis in an autocrine fashion in ARDS was tested using cultured endothelial cells stimulated with MIF and a murine model of lipopolysaccharide ( LPS ) -induced acute lung injury . MIF induced significant MIF and tumour necrosis factor ( P01375 ) -alpha synthesis in cultured endothelial cells and the effect was blocked by neutralizing anti-MIF antibody . A similar blocking effect was observed when MIF-stimulated endothelial cells were pretreated with neutralizing anti- P01375 antibody or glucocorticoid , supporting the notion that MIF induced P01375 production via an amplifying pro-inflammatory loop . Treatment with anti-MIF or glucocorticoid effectively attenuated pulmonary pathology and the synthesis of MIF or P01375 in mice with LPS-induced acute lung injury . Mildly augmented expression of aquaporin 1 ( P29972 ) was also detected in alveolar capillary endothelium in ARDS . In vitro studies revealed that both MIF and P01375 induced a small increase of P29972 synthesis in cultured endothelial cells . These findings suggest that MIF plays a crucial pathological role leading to alveolar inflammation in ARDS . Anti-MIF and early glucocorticoid therapy may represent a novel therapeutic approach for reducing alveolar inflammation in ARDS .

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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 . 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 . DB08890 -a novel secretagogue in the treatment of irritable bowel syndrome with constipation and chronic idiopathic constipation . Irritable bowel syndrome with constipation ( IBS-C ) and chronic idiopathic constipation ( Q96RK0 ) are highly prevalent gastrointestinal disorders . Traditional symptoms based therapies had somewhat limited success and efficacy in addressing the disorders . Recently , linaclotide emerged as novel peptide capable of improving abdominal symptoms in patients suffering from IBS-C and Q96RK0 . Guanylate cyclase C ( P25092 ) receptor a multi domain protein , found to be molecular target for linaclotide which acts by activating P25092 receptor on the apical surface of intestinal epithelial cells . Binding of linaclotide to P25092 receptor triggers the elevation of second messenger cGMP that elicits fluid secretion into intestinal cells which play a critical role in maintaining homeostasis through cystic fibrosis transmembrane conductance regulator ( P13569 ) . Data from Phase II and III clinical trials demonstrated that linaclotide seems to produce a statistically significant increase in stool frequency , improved straining , decreased abdominal pain and discomfort . 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 . The role of vascular endothelial growth factor and estradiol in the regulation of endometrial angiogenesis and cell proliferation in the marmoset . The present studies explore the roles of vascular endothelial growth factor ( P15692 ) and estradiol on angiogenesis and stromal and epithelial cell proliferation in the marmoset endometrium during the proliferative phase of the ovulatory cycle . At the start of the proliferative phase , marmosets were 1 ) treated with vehicle , 2 ) treated with a P15692 inhibitor ( DB08885 , aflibercept ) , 3 ) ovariectomized , 4 ) ovariectomized and given replacement estradiol , or 5 ) treated with DB08885 and given replacement estradiol . The uterus was examined 10 d later in the late proliferative phase . Changes in endothelial and epithelial cell proliferation were quantified using a volumetric density method after immunohistochemistry for bromodeoxyuridine to localize proliferating cells , CD31 to visualize endothelial cells , and dual staining to distinguish endothelial cell proliferation . Endothelial proliferation was elevated in late proliferative controls but virtually absent after DB08885 . Ovariectomy had a similar inhibitory effect , whereas angiogenesis was restored by estrogen replacement . Estradiol replacement in DB08885 -treated marmosets resulted in only a small increase in endothelial cell proliferation that remained significantly below control values . DB08885 treatment and ovariectomy also markedly reduced stromal cell proliferation but resulted in increased stromal cell density associated with a reduction in overall endometrial volume . Estrogen replacement in both ovariectomized and DB08885 -treated animals restored stromal proliferation rates and cell density . These results show that endometrial angiogenesis and stromal proliferation during the proliferative phase are driven by estradiol and that the effect of estrogen on angiogenesis is mediated largely by P15692 . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . P12318 polymorphism is correlated with clinical outcome after immunotherapy of neuroblastoma with anti-GD2 antibody and granulocyte macrophage colony-stimulating factor . PURPOSE : Anti-GD2 murine IgG3 antibody 3F8 kills neuroblastoma cells by antibody-dependent cell-mediated cytotoxicity ( ADCC ) . Granulocyte macrophage colony-stimulating factor ( GM- P04141 ) enhances phagocyte-mediated ADCC . The differential affinity of the human FCGR polymorphic alleles for 3F8 may influence the effectiveness of antibody immunotherapy . PATIENTS AND METHODS : The entire cohort of high risk neuroblastoma patients ( N = 136 ) treated on protocol using 3F8 and GM- P04141 were the subjects of this analysis . Tumor response was measured by standard clinical tools plus sensitive molecular monitoring using quantitative reverse transcription-polymerase chain reaction ( qRT-PCR ) . Polymorphic alleles of P12318 and P08637 were determined by PCR plus direct sequencing using genomic DNA samples obtained from marrow or blood of patients . RESULTS : P12318 ( R/R ) genotype correlated with progression-free survival for the entire cohort ( P = .049 ) and for the subset of patients with no history of prior relapse ( P = .023 ) . P12318 ( R/R ) also correlated with marrow remission 2.5 months after treatment initiation : by histology ( P = .021 and P = .036 , for the entire cohort and the subset , respectively ) and by qRT-PCR ( P = .052 and P = .033 , respectively ) . CONCLUSION : The favorable outcome associated with P12318 ( R/R ) genotype is consistent with the proposed role of P12318 and phagocyte-mediated ADCC in 3F8 plus GM- P04141 immunotherapy . Antihistamine effects on prefrontal cortex activity during working memory process in preschool children : a near-infrared spectroscopy ( NIRS ) study . P35367 antagonists ( antihistamines ) are widely used for the treatment of allergic disorders in young children . This study examined the effects of antihistamine on prefrontal cortex activity in preschool children using near-infrared spectroscopy ( NIRS ) , an emerging brain-imaging method suitable for psychological experiments , especially in young children . We examined the changes of oxygenated hemoglobin concentration in the prefrontal cortex while children performed a spatial working memory task , 3h after taking a first-generation antihistamine ( ketotifen ) , second-generation antihistamine ( epinastine ) , or placebo . Fifteen healthy preschool children ( mean age , 5.5 years ) participated . DB00920 significantly impaired behavioral performance and cortical activation at the lateral prefrontal cortex in the working memory task , compared with epinastine and placebo . There were no sedative effects on neural response or behavioral performance after epinastine administration . This paper demonstrates for the first time differential sedation effects of first- and second-generation antihistamines on brain hemodynamic response in young children . Also discussed is the utility of the NIRS technique in neuropsychopharmacological studies of children . 5- Q13049 receptor induces P29323 phosphorylation and proliferation through ADAM-17 tumor necrosis factor-alpha-converting enzyme ( P78536 ) activation and heparin-bound epidermal growth factor-like growth factor ( HB- P01133 ) shedding in mesangial cells . In this study , we present multiple lines of evidence to support a critical role for heparin-bound P01133 ( epidermal growth factor ) -like growth factor ( HB- P01133 ) and tumor necrosis factor-alpha-converting enzyme ( P78536 ) ( P78536 ) in the transactivation of P01133 receptor ( P00533 ) , P29323 phosphorylation , and cellular proliferation induced by the 5-HT(2A) receptor in renal mesangial cells . 5-hydroxy-tryptamine ( 5-HT ) resulted in rapid activation of P78536 , HB- P01133 shedding , P00533 activation , P29323 phosphorylation , and longer term increases in DNA content in mesangial cells . P29323 phosphorylation was attenuated by 1 ) neutralizing P00533 antibodies and the P00533 kinase inhibitor , AG1478 , 2 ) neutralizing HB- P01133 , but not amphiregulin , antibodies , heparin , or CM197 , and 3 ) pharmacological inhibitors of matrix-degrading metalloproteinases or P78536 small interfering RNA . Exogenously administered HB- P01133 stimulated P29323 phosphorylation . Additionally , P78536 was co-immunoprecipitated with HB- P01133 . Small interfering RNA against P78536 also blocked 5-HT-induced increases in P29323 phosphorylation , HB- P01133 shedding , and DNA content . In aggregate , this work supports a pathway map that can be depicted as follows : 5-HT --> 5-HT(2A) receptor --> P78536 --> HB- P01133 shedding --> P00533 --> P29323 --> increased DNA content . To our knowledge , this is the first time that P78536 has been implicated in 5-HT-induced P00533 transactivation or in proliferation induced by a G protein-coupled receptor in native cells in culture . Further characterization of a somatic cell hybrid panel : ten new assignments to the bovine genome . Thirty-six partially characterized hamster-bovine hybrid cell lines were used for the determination of synteny groups . Sixteen additional reference loci , selected for their coverage of the bovine genome , were analysed on these hybrid cells . This increases to 25 the number of synteny groups detected . This panel was then used to make synteny assignments for 10 additional loci , eight by Southern blotting ( P02452 , P08123 , FAS , P07858 , P07711 , P07510 , P07686 and P08908 ) and two by polymerase chain reaction ( PCR ) amplification ( P35367 and ETH1112 ) . These loci were assigned to international synteny groups U12 ( P35367 ) , U13 ( P08123 ) , U17 ( P07510 ) , U21 ( P02452 , FAS ) , U29 ( ETH1112 ) , to chromosome 20 ( U14 or U25 ) for P07686 and P08908 , and to the same local synteny group ( A ) , which is probably U18 , for P07858 and P07711 . For three loci already mapped in humans ( P02452 , P08123 and P07510 ) , the present results are in accordance with the predictions based on comparative mapping between the human and bovine species . Effects of short- and long-term risperidone treatment on prolactin levels in children with autism . BACKGROUND : The effects of short- and long-term risperidone treatment on serum prolactin were assessed in children and adolescents with autism . METHODS : Patients with autism ( N = 101 , 5-17 years of age ) were randomized to an 8-week trial of risperidone or placebo and 63 then took part in a 4-month open-label follow-up phase . Serum samples were obtained at Baseline and Week-8 ( N = 78 ) , and at 6-month ( N = 43 ) and 22-month ( N = 30 ) follow-up . Serum prolactin was determined by immunoradiometric assay ; dopamine type-2 receptor ( P14416 ) polymorphisms were genotyped . RESULTS : Baseline prolactin levels were similar in the risperidone ( N = 42 ) and placebo ( N = 36 ) groups ( 9.3 +/- 7.5 and 9.3 +/- 7.6 ng/ml , respectively ) . After 8 weeks of risperidone , prolactin increased to 39.0 +/- 19.2 ng/ml , compared with 10.1 +/- 8.8 ng/ml for placebo ( p < .0001 ) . P01236 levels were also significantly increased at 6 months ( 32.4 +/- 17.8 ng/ml ; N = 43 , p < .0001 ) and at 22 months ( N = 30 , 25.3 +/- 15.6 ng/ml , p < .0001 ) . P01236 levels were not associated with adverse effects and P14416 alleles ( Taq1A , -141C Ins/Del , C957T ) did not significantly influence baseline levels or risperidone-induced increases in prolactin . CONCLUSIONS : DB00734 treatment was associated with two- to four-fold mean increases in serum prolactin in children with autism . Although risperidone-induced increases tended to diminish with time , further research on the consequences of long-term prolactin elevations in children and adolescents is needed . Monoclonal antibodies against P00533 in non-small cell lung cancer . Blockade of the epidermal growth factor receptor ( P00533 ) by monoclonal antibodies is a strategy to improve outcome in patients with non-small cell lung cancer . Cetuximab , a chimeric anti- P00533 monoclonal antibody , has been studied in combination with different chemotherapy protocols in both phase II and phase III trials in patients with advanced NSCLC . In the phase III FLEX trial , cetuximab added to cisplatin/vinorelbine resulted in an absolute overall survival benefit of 1.2 months compared to the same chemotherapy alone in patients with advanced P00533 -expressing NSCLC . In the second phase III trial , cetuximab added to carboplatin plus paclitaxed failed to improve progression-free survival but suggested a survival benefit similar to that seen in the FLEX trial . However , the benefit in survival reached statistical significance only in the FLEX trial . A meta-analysis that included patients from four randomized trials confirmed the efficacy of cetuximab when added to chemotherapy . Thus addition of cetuximab to platinum-based chemotherapy represents a new treatment option for patients with advanced NSCLC . DB05101 and panitumumab have also been evaluated in phase II trials . DB09559 is currently evaluated in combination with chemotherapy in two randomized phase III trials . 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 . 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 . 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 . Effects of salinity and prolactin on gene transcript levels of ion transporters , ion pumps and prolactin receptors in Mozambique tilapia intestine . Euryhaline teleosts are faced with significant challenges during changes in salinity . Osmoregulatory responses to salinity changes are mediated through the neuroendocrine system which directs osmoregulatory tissues to modulate ion transport . P01236 ( PRL ) plays a major role in freshwater ( FW ) osmoregulation by promoting ion uptake in osmoregulatory tissues , including intestine . We measured mRNA expression of ion pumps , Na(+)/K(+)-ATPase α3-subunit ( NKAα3 ) and vacuolar type H(+)-ATPase A-subunit ( V-ATPase A-subunit ) ; ion transporters/channels , Na(+)/K(+)/2Cl(-) co-transporter ( Q13621 ) and cystic fibrosis transmembrane conductance regulator ( P13569 ) ; and the two PRL receptors , PRLR1 and PRLR2 in eleven intestinal segments of Mozambique tilapia ( Oreochromis mossambicus ) acclimated to FW or seawater ( SW ) . Gene expression levels of NKAα3 , V-ATPase A-subunit , and Q13621 were generally lower in middle segments of the intestine , whereas P13569 mRNA was most highly expressed in anterior intestine of FW-fish . In both FW- and SW-acclimated fish , PRLR1 was most highly expressed in the terminal segment of the intestine , whereas PRLR2 was generally most highly expressed in anterior intestinal segments . While Q13621 , NKAα3 and PRLR2 mRNA expression was higher in the intestinal segments of SW-acclimated fish , P13569 mRNA expression was higher in FW-fish ; PRLR1 and V-ATPase A-subunit mRNA expression was similar between FW- and SW-acclimated fish . Next , we characterized the effects of hypophysectomy ( Hx ) and PRL replacement on the expression of intestinal transcripts . Hypophysectomy reduced both Q13621 and P13569 expression in particular intestinal segments ; however , only Q13621 expression was restored by PRL replacement . Together , these findings describe how both acclimation salinity and PRL impact transcript levels of effectors of ion transport in tilapia intestine . 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 . Promoter methylation status of P15692 receptor genes : a possible epigenetic biomarker to anticipate the efficacy of intracellular-acting P15692 -targeted drugs in cancer cells . We evaluated whether the inhibitory effects of vascular endothelial growth factor ( P15692 ) -targeted drugs on the proliferation of cancer cells differed according to P15692 receptor ( VEGFR ) genes , Flt1 and P35968 , promoter methylation status . Five hyper-VEGFR-methylation and six no-VEGFR-methylation cancer cells were used for the present study , together with human umbilical endothelial cells ( HUVECs ) as a control . No-VEGFR-methylation cancer cells showed higher expression of Flt1 and P35968 than hyper-VEGFR-methylation cancer cells . Hyper-VEGFR-methylation cancer cells only showed increased expression and protein levels of Flt1 and P35968 after treatment with the demethylase DB01262 . Two drugs ( a P15692 -specific-antibody , bevacizumab , and a P35968 -specific-antibody ) targeting extracellular P15692 -VEGFR signaling and two P15692 -specific-tyrosine kinase inhibitors ( DB04879 and sunitinib ) targeting intracellular VEGFR signaling were used in the cell proliferation assay . HUVECs showed dose- and time-dependent proliferation decrease with all tested drugs over a 72 h incubation period . No- or hyper-VEGFR-methylation cancer cells showed no significant proliferation differences after treatment with P15692 -specific-antibody or P35968 -specific-antibody . After DB04879 or sunitinib treatment , no-VEGFR-methylation cancer cells showed dose- or time-dependent decreases in proliferation . Hyper-VEGFR-methylation cancer cells also showed proliferation inhibition by P15692 -specific-tyrosine kinase inhibitors after demethylation of Flt1 and P35968 . Proliferation inhibition synergistically increased after combination of demethylation with DB04879 in hyper- P15692 -methylation cancer cells . We observed that intracellular targeting of P15692 -VEGFR signaling could be more effective than extracellular targeting of the pathway in the suppression of proliferation of some cancer cells . In particular , the efficacy of intracellular targeting of P15692 -specific-tyrosine kinase inhibitors might be influenced by the epigenetic alteration of VEGFRs . Clinical endocannabinoid deficiency ( CECD ) : can this concept explain therapeutic benefits of cannabis in migraine , fibromyalgia , irritable bowel syndrome and other treatment-resistant conditions ? OBJECTIVES : This study examines the concept of clinical endocannabinoid deficiency ( CECD ) , and the prospect that it could underlie the pathophysiology of migraine , fibromyalgia , irritable bowel syndrome , and other functional conditions alleviated by clinical cannabis . METHODS : Available literature was reviewed , and literature searches pursued via the National Library of Medicine database and other resources . RESULTS : Migraine has numerous relationships to endocannabinoid function . Anandamide ( AEA ) potentiates P08908 and inhibits 5- Q13049 receptors supporting therapeutic efficacy in acute and preventive migraine treatment . Cannabinoids also demonstrate dopamine-blocking and anti-inflammatory effects . AEA is tonically active in the periaqueductal gray matter , a migraine generator . THC modulates glutamatergic neurotransmission via DB01221 receptors . Fibromyalgia is now conceived as a central sensitization state with secondary hyperalgesia . Cannabinoids have similarly demonstrated the ability to block spinal , peripheral and gastrointestinal mechanisms that promote pain in headache , fibromyalgia , IBS and related disorders . The past and potential clinical utility of cannabis-based medicines in their treatment is discussed , as are further suggestions for experimental investigation of CECD via P04141 examination and neuro-imaging . CONCLUSION : Migraine , fibromyalgia , IBS and related conditions display common clinical , biochemical and pathophysiological patterns that suggest an underlying clinical endocannabinoid deficiency that may be suitably treated with cannabinoid medicines . DB00073 -dependent cytotoxicity by natural killer cells : influence of P08637 polymorphism on the concentration-effect relationship . The P08637 gene dimorphism generates two allotypes : FcgammaRIIIa-158V and FcgammaRIIIa-158F . The genotype homozygous for FcgammaRIIIa-158V ( VV ) is associated with higher clinical response to rituximab , a chimeric anti- P11836 IgG1 used in the treatment of B lymphoproliferative malignancies . Our objective was to determine whether this genetic association relates to rituximab-dependent cytotoxicity mediated by FcgammaRIIIa/CD16a+ cells . The number of CD16+ circulating monocytes , T cells , and natural killer ( NK ) cells in 54 donors was first shown to be unrelated to P08637 polymorphism . We then demonstrated that FcgammaRIIIa-158V displays higher affinity for rituximab than FcgammaRIIIa-158F by comparing rituximab concentrations inhibiting the binding of 3G8 mAb ( anti-CD16 ) with VV NK cells and NK cells homozygous for FcgammaRIIIa-158F ( FF ) . VV and FF NK cells killed Daudi cells similarly after FcgammaRIIIa engagement by saturating concentrations of rituximab or 3G8 . However , the rituximab concentration resulting in 50 % lysis ( EC(50) ) observed with NK cells from VV donors was 4.2 times lower than that observed with NK cells from FF donors ( on average 0.00096 and 0.00402 microg/ml , respectively , P = 0.0043 ) . Finally , the functional difference between VV and FF NK cells was restricted to rituximab concentrations weakly sensitizing P11836 . This study supports the conclusion that P08637 genotype is associated with response to rituximab because it affects the relationship between rituximab concentration and NK cell-mediated lysis of P11836 + cells . DB00073 administration could therefore be adjusted according to P08637 genotype .

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interacts_with DB01211?

multiple_choice

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 . 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 . 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 . DB00205 and WR99210 exert opposing selection on dihydrofolate reductase from Plasmodium vivax . Plasmodium vivax is a major public health problem in Asia and South and Central America where it is most prevalent . Until very recently , the parasite has been effectively treated with chloroquine , but resistance to this drug has now been reported in several areas . Affordable alternative treatments for vivax malaria are urgently needed . DB00205 -sulfadoxine is an inhibitor of dihydrofolate reductase ( P00374 ) that has been widely used to treat chloroquine-resistant Plasmodium falciparum malaria . P00374 inhibitors have not been considered for treatment of vivax malaria , because initial trials showed poor efficacy against P. vivax . P. vivax can not be grown in culture ; the reason for its resistance to P00374 inhibitors is unknown . We show that , like P. falciparum , point mutations in the dhfr gene can cause resistance to pyrimethamine in P. vivax . WR99210 is a novel inhibitor of P00374 , effective even against the most pyrimethamine-resistant P. falciparum strains . We have found that it is also an extremely effective inhibitor of the P. vivax P00374 , and mutations that confer high-level resistance to pyrimethamine render the P. vivax enzyme exquisitely sensitive to WR99210 . These data suggest that pyrimethamine and WR99210 would exert opposing selective forces on the P. vivax population . If used in combination , these two drugs could greatly slow the selection of parasites resistant to both drugs . If that is the case , this novel class of P00374 inhibitors could provide effective and affordable treatment for chloroquine- and pyrimethamine-resistant vivax and falciparum malaria for many years to come . Development of DB00644 antagonists for prostate cancer : new approaches to treatment . Prostate cancer has become the most common cancer among American men and is second only to lung cancer as a cause of male cancer-related death . Several treatment options exist for different stages of prostate cancer including observation , prostatectomy , radiation therapy , chemotherapy , and hormone therapy . Hormone therapy has evolved from the use of estrogens to gonadotropin-releasing hormone ( DB00644 ) agonists and recently , investigational DB00644 antagonists . P30968 agonists such as leuprolide , bruserelin and goserelin have been used for the treatment of prostate cancer . These agonists eventually cause the inhibition of lutenizing hormone production , which in turn causes a suppression of testosterone and dihydrotestosterone , on which continued growth of prostate cancer cells depend . Several comparative studies of leuprorelin administered as daily injections or monthly depot injections have been reported . Disease progression was prevented in more than 72 % of men administered daily leuprorelin , and in 82 % to 89 % of those receiving monthly depots . Another synthetic DB00644 analog , goserelin , has been studied in a similar population of men with daily injections producing partial responses in 60 % to 80 % of men with previously untreated prostate cancer . DB00106 , a peptide antagonist of P30968 , is also being studied for the treatment of prostate cancer . The discovery and development of DB00644 antagonists may provide an important advance for patients with prostate cancer . Clearly the studies described herein , as well as many others , outline an exciting era of research to define the optimal use of hormonal therapy in prostate cancer . Hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in the rat . Growth hormone-releasing peptides ( Q92847 ) and ghrelin are synthetic and natural ligands of growth hormone secretagogue receptor ( Q92847 ) respectively and are shown to exert protective actions on cardiac dysfunction . Because ghrelin has been reported to inhibit proinflammatory responses in human endothelium and Q92847 has been identified in blood vessels , we hypothesized that Q92847 could alleviate the development of atherosclerosis ( As ) . Atherosclearosis was induced by a short period ( 4 days ) of vitamin D(3) and chronic ( three months ) intragastric feeding of high fat emulsion ( containing 0.5 % propylthiouracil ) in adult SD rats . Some As rats received chronic hexarelin ( a variant of Q92847 ) injection ( SC P55957 , 30 days ) and normal rats received placebo as control . Significant atherosclerosis developed in animals fed with the emulsion . Serum total cholesterol and LDL-c increased , and HDL-c and aortic nitric oxide ( NO ) decreased significantly in As group . Hexarelin suppressed the formation of atherosclerotic plaques and neointima , partially reversed serum HDL-c/LDL-c ratio and increased the levels of serum NO and aortic mRNAs of P29474 , Q92847 and P16671 in As rats . Hexarelin also decreased [ (3)H ] -TdR incorporation in cultured vascular smooth muscle cell ( VSMC ) and calcium sedimentation in aortic wall . Furthermore , foam cell formation induced by ox-LDL was decreased by hexarelin . In conclusion , hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in rats , possibly through upregulating HDL-c/LDL-c ratio , vascular NO production and downregulating the VSMC proliferation , aortic calcium sedimentation and foam cell formation . These novel anti-atherosclerotic actions of hexarelin suggest that the peptide might have a clinical potential in treating atherosclerosis . 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 . 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 . Characterization of the inhibitory effects of erythromycin and clarithromycin on the Q12809 potassium channel . Both erythromycin and clarithromycin have been reported to cause QT prolongation and the cardiac arrhythmia torsade de pointes in humans , however direct evidence documenting that these drugs produce this effect by blocking human cardiac ion channels is lacking . The goal of this study was to test the hypothesis that these macrolide antibiotics significantly block the delayed rectifier current ( IKr ) encoded by Q12809 ( the human ether-a-go-go-related gene ) at drug concentrations , temperature and ionic conditions mimicking those occurring in human subjects . DB01345 currents in P29320 293 cells stably transfected with Q12809 were recorded using a whole cell voltage clamp method . Exposure of cells to erythromycin reduced the Q12809 encoded potassium current in a concentration dependent manner with an IC50 of 38.9 +/- 1.2 microM and Hill Slope factor of 0.4 +/- 0.1 . DB01211 produced a similar concentration-dependent block with an IC50 of 45.7 +/- 1.1 microM and Hill Slope factor of 1.0 +/- 0.1 . Erythromycin ( 25-250 microM ) and clarithromycin ( 5 or 25 microM ) also produced a significant decrease in the integral of the current evoked by an action potential shaped voltage clamp protocol . The results of this study document that both erythromycin and clarithromycin significantly inhibit the Q12809 potassium current at clinically relevant concentrations . Preserving mitochondrial function prevents the proteasomal degradation of P30793 . The development of pulmonary hypertension is a common accompaniment of congenital heart disease ( Q8NE62 ) with increased pulmonary blood flow . Our recent evidence suggests that asymmetric dimethylarginine ( DB01686 ) -induced mitochondrial dysfunction causes endothelial nitric oxide synthase ( P29474 ) uncoupling secondary to a proteasome-dependent degradation of P30793 ( P30793 ) that results in a decrease in the NOS cofactor tetrahydrobiopterin ( BH(4) ) . Decreases in NO signaling are thought to be an early hallmark of endothelial dysfunction . As l-carnitine plays an important role in maintaining mitochondrial function , in this study we examined the protective mechanisms and the therapeutic potential of l-carnitine on NO signaling in pulmonary arterial endothelial cells and in a lamb model of Q8NE62 and increased pulmonary blood flow ( Shunt ) . Acetyl-l-carnitine attenuated the DB01686 -mediated proteasomal degradation of P30793 . This preservation was associated with a decrease in the association of P30793 with Hsp70 and the C-terminus of Hsp70-interacting protein ( Q9UNE7 ) and a decrease in its ubiquitination . This in turn prevented the decrease in BH(4) levels induced by DB01686 and preserved NO signaling . Treatment of Shunt lambs with l-carnitine also reduced P30793 / Q9UNE7 interactions , attenuated the ubiquitination and degradation of P30793 , and increased BH(4) levels compared to vehicle-treated Shunt lambs . The increases in BH(4) were associated with decreased NOS uncoupling and enhanced NO generation . Thus , we conclude that L-carnitine may have a therapeutic potential in the treatment of pulmonary hypertension in children with Q8NE62 with increased pulmonary blood flow . 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 . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . 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 . The human gonadotropin releasing hormone type I receptor is a functional intracellular GPCR expressed on the nuclear membrane . The mammalian type I gonadotropin releasing hormone receptor ( P30968 ) is a structurally unique G protein-coupled receptor ( GPCR ) that lacks cytoplasmic tail sequences and displays inefficient plasma membrane expression ( PME ) . Compared to its murine counterparts , the primate type I receptor is inefficiently folded and retained in the endoplasmic reticulum ( ER ) leading to a further reduction in PME . The decrease in PME and concomitant increase in intracellular localization of the mammalian DB00644 -RI led us to characterize the spatial distribution of the human and mouse DB00644 receptors in two human cell lines , P29320 293 and HTR-8/SVneo . In both human cell lines we found the receptors were expressed in the cytoplasm and were associated with the ER and nuclear membrane . A molecular analysis of the receptor protein sequence led us to identify a putative monopartite nuclear localization sequence ( NLS ) in the first intracellular loop of DB00644 -RI . Surprisingly , however , neither the deletion of the NLS nor the addition of the Xenopus P30968 cytoplasmic tail sequences to the human receptor altered its spatial distribution . Finally , we demonstrate that DB00644 treatment of nuclei isolated from P29320 293 cells expressing exogenous DB00644 -RI triggers a significant increase in the acetylation and phosphorylation of histone H3 , thereby revealing that the nuclear-localized receptor is functional . Based on our findings , we conclude that the mammalian DB00644 -RI is an intracellular GPCR that is expressed on the nuclear membrane . This major and novel discovery causes us to reassess the signaling potential of this physiologically and clinically important receptor . Intercellular adhesion molecule-1 ( P05362 ) expression and soluble P05362 ( sICAM-1 ) production by cytokine-activated human aortic endothelial cells : a possible role for P05362 and sICAM-1 in atherosclerotic aortic aneurysms . The interactions of inflammatory cells , cytokines , and cell adhesion molecules ( P62158 ) may be important in the pathogenesis of vascular diseases such as abdominal aortic aneurysms ( AAA ) , in which inflammation plays a role . The aim of this study was to investigate the pathogenic role of P05362 , a molecule involved in leucocyte-endothelial interactions , in vascular inflammation . ELISA of human explant culture supernatants revealed a four-fold increase in sICAM-1 production by AAA ( n = 9 ) versus normal ( n = 8 ) aortic explants . Human aortic endothelial cell ( hAEC ) culture was used for further studies as an in vitro model for aortic inflammatory conditions . Tumour necrosis factor-alpha ( P01375 ) or P01584 treatment of hAEC resulted in an up to 1.8-fold significant increase in sICAM-1 production compared with resting cells . In addition , the expression of P05362 on cytokine-stimulated versus resting hAEC was measured by radioimmunoassay . P01375 significantly induced P05362 expression on these cells . These results suggest that different forms of P05362 , present on or released by the activated aortic endothelium , may be involved in leucocyte adhesion to and migration into the vessel wall . 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 . 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 . [ Expressions of receptor tyrosine kinases mRNA and protein in carcinoma of bladder ] . OBJECTIVE : To detect the expressions of receptor tyrosine kinases ( RTKs ) mRNA and protein and to explore potentially promising tumor markers and conceivable drug target in bladder cancer . METHODS : The expressions of RTKs mRNA and protein in tissue from invasive urothelial carcinoma of the bladder were examined by real-time quantitative PCR array and cytokine antibody array , with normal bladder tissue as control . The Results were analyzed using bioinformatic approaches . RESULTS : The expressions of P01135 , Q9NY15 , P05121 , O15123 , Q9NQ38 , O95841 , P58294 , P21741 , Q07325 , P28799 , Q01196 , P15692 , and P01137 were obviously upregulated in bladder cancer tissue , while those of O43854 , P21246 , P13500 , Q9GZP0 , Q92913 , P21583 , P09038 , P36955 , and P01375 were downregulated . Q9UM73 , Btk , EphB2 , ErbB4 , P09619 -α , ROS , Tie-2 , Tyk2 , and P35916 were over-expressed in bladder cancer , while P42685 , Fyn , IGF-IR , P01308 R , Itk , P23458 , P52333 , and P06239 were low-expressed . CONCLUSION : Vascular endothelial growth factor/platelet-derived growth factor-targeted therapies may play an active role in treating carcinoma of bladder . [ 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_1568

MH_train_1568

MH_train_1568

interacts_with DB00072?

multiple_choice

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 . 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 . Inhibition of the leukotriene synthetase of rat basophil leukemia cells by diethylcarbamazine , and synergism between diethylcarbamazine and piriprost , a P09917 inhibitor . DB00711 inhibited the formation of sulfidopeptide leukotrienes in rat basophil leukemia ( RBL ) cells ( 50 % inhibitory concentration , EC50 , 3 mM ) . Similar concentrations also inhibited the formation of leukotriene C4 ( LTC4 ) by LTC synthetase , a detergent-solubilized cell free particulate enzyme from RBL cells which is capable of coupling P09960 to glutathione . By contrast , the conversion of P09960 to LTC4 using enzymes from rat liver was at least ten times less sensitive to this inhibitor . The EC50 for inhibition of the leukotriene C synthetase of RBL cells was directly proportional to the P09960 concentration in the incubations , ranging from 1.5 mM at 10 microM P09960 to over 40 mM at 500 microM P09960 . Kinetic analysis revealed that the inhibition of the leukotriene C synthetase reaction by diethylcarbamazine was competitive with respect to P09960 . In contrast to diethylcarbamazine , piriprost ( U-60,257 ; 6,9-deepoxy-6,9-(phenylimino)-delta 6,8-prostaglandin I1 ) , which inhibits the formation of sulfidopeptide leuktrienes in RBL cells at the P09917 step ( EC50 5 microM ) , did not inhibit the leukotriene synthetase of these cells . On the other hand , low concentrations of piriprost , which had no demonstrable inhibitory activity on leukotriene formation by themselves , markedly synergized the inhibitory activity of diethylcarbamazine . These results are consistent with the interpretation that both piriprost and diethylcarbamazine inhibit leukotriene formation but that they act on sequential steps in the biosynthetic pathway in such a manner as to synergistically interfere with the availability or utilization of P09960 in the leukotriene C synthetase reaction . OSU-A9 , a potent indole-3-carbinol derivative , suppresses breast tumor growth by targeting the Akt-NF-kappaB pathway and stress response signaling . The molecular heterogeneity of human tumors challenges the development of effective preventive and therapeutic strategies . To overcome this issue , a rational approach is the concomitant targeting of clinically relevant cellular abnormalities with combination therapy or a potent multi-targeted agent . OSU-A9 is a novel indole-3-carbinol derivative that retains the parent compound 's ability to perturb multiple components of oncogenic signaling , but provides marked advantages in chemical stability and antitumor potency . Here , we show that OSU-A9 exhibits two orders of magnitude greater potency than indole-3-carbinol in inducing apoptosis in various breast cancer cell lines with distinct genetic abnormalities , including MCF-7 , MDA-MB-231 and SKBR3 , with the half maximal inhibitory concentration in the range of 1. P35326 .8 microM vis-à-vis 200 microM for indole-3-carbinol . This differential potency was paralleled by OSU-A9 's superior activity against multiple components of the Akt-nuclear factor-kappa B ( NF-kappaB ) and stress response signaling pathways . Notable among these were the increased estrogen receptor ( ER ) -beta/ERalpha expression ratio , reduced expression of P04626 and P61073 and the upregulation of aryl hydrocarbon receptor expression and its downstream target NF-E2 P29466 -regulated factor ( Nrf2 ) . Non-malignant MCF-10A cells were resistant to OSU-A9 's antiproliferative effects . Daily oral administration of OSU-A9 at 25 and 50 mg/kg for 49 days significantly inhibited MCF-7 tumor growth by 59 and 70 % , respectively , without overt signs of toxicity or evidence of induced hepatic biotransformation enzymes . In summary , OSU-A9 is a potent , orally bioavailable inhibitor of the Akt-NF-kappaB signaling network , targeting multiple aspects of breast tumor pathogenesis and progression . Thus , its translational potential for the treatment or prevention of breast cancer warrants further investigation . Clonal differences in expression of 25-hydroxyvitamin D(3)-1alpha-hydroxylase , of 25-hydroxyvitamin D(3)-24-hydroxylase , and of the vitamin D receptor in human colon carcinoma cells : effects of epidermal growth factor and 1alpha,25-dihydroxyvitamin D(3) . Human colon carcinoma cells express 25-hydroxyvitamin D(3)-1alpha-hydroxylase ( O15528 ) and thus produce the vitamin D receptor ( P11473 ) ligand 1alpha,25-dihydroxyvitamin D(3) ( 1,25-D3 ) , which can be metabolized by 25-hydroxyvitamin D(3)-24-hydroxylase ( Q07973 ) . Expression of P11473 , O15528 , and Q07973 determines the efficacy of the antimitotic action of 1,25-D3 and is distinctly related to the degree of differentiation of cancerous lesions . In the present study we addressed the question of whether the effects of epidermal growth factor ( P01133 ) and of 1,25-D3 on P11473 , O15528 , and Q07973 gene expression in human colon carcinoma cell lines also depend on the degree of cellular differentiation . We were able to show that slowly dividing , highly differentiated Caco-2/15 cells responded in a dose-dependent manner to both P01133 and 1,25-D3 by up-regulation of P11473 and O15528 expression , whereas in highly proliferative , less differentiated cell lines , such as Caco-2/AQ and COGA-1A and -1E , negative regulation was observed . Q07973 mRNA was inducible in all clones by 1,25-D3 but not by P01133 . From the observed clonal differences in the regulatory effects of P01133 and 1,25-D3 on P11473 and O15528 gene expression we suggest that P11473 -mediated growth inhibition by 1,25-D3 would be efficient only in highly differentiated carcinomas even when under mitogenic stimulation by P01133 . Vitamin D metabolism , mechanism of action , and clinical applications . DB00169 is made in the skin from 7-dehydrocholesterol under the influence of UV light . DB00153 ( ergocalciferol ) is derived from the plant sterol ergosterol . Vitamin D is metabolized first to 25 hydroxyvitamin D ( 25OHD ) , then to the hormonal form 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . Q6VVX0 is the most important 25-hydroxylase ; O15528 is the key 1-hydroxylase . Both 25OHD and 1,25(OH)2D are catabolized by Q07973 . 1,25(OH)2D is the ligand for the vitamin D receptor ( P11473 ) , a transcription factor , binding to sites in the DNA called vitamin D response elements ( VDREs ) . There are thousands of these binding sites regulating hundreds of genes in a cell-specific fashion . P11473 -regulated transcription is dependent on comodulators , the profile of which is also cell specific . Analogs of 1,25(OH)2D are being developed to target specific diseases with minimal side effects . This review will examine these different aspects of vitamin D metabolism , mechanism of action , and clinical application . Anti-ErbB-2 mAb therapy requires type I and II interferons and synergizes with anti- P18621 or anti- Q07011 mAb therapy . DB00072 , a monoclonal antibody targeting human epidermal growth factor receptor-2 ( P04626 /ErbB-2 ) , has become the mainstay of treatment for P04626 -positive breast cancer . Nevertheless , its exact mechanism of action has not been fully elucidated . Although several studies suggest that Fc receptor-expressing immune cells are involved in trastuzumab therapy , the relative contribution of lymphocyte-mediated cellular cytotoxicity and antitumor cytokines remains unknown . We report here that anti-ErbB-2 mAb therapy is dependent on the release of type I and type II IFNs but is independent of perforin or P48023 . Our study thus challenges the notion that classical antibody-dependent , lymphocyte-mediated cellular cytotoxicity is important for trastuzumab . We demonstrate that anti-ErbB-2 mAb therapy of experimental tumors derived from MMTV-ErbB-2 transgenic mice triggers MyD88-dependent signaling and primes IFN-γ-producing CD8+ T cells . Adoptive cell transfer of purified T cell subsets confirmed the essential role of IFN-γ-producing CD8+ T cells . Notably , anti-ErbB-2 mAb therapy was independent of IL-1R or IL-17Ra signaling . Finally , we investigated whether immunostimulatory approaches with antibodies against programmed death-1 ( P18621 ) or 41BB ( Q07011 ) could be used to capitalize on the immune-mediated effects of trastuzumab . We demonstrate that anti- P18621 or anti- Q07011 mAb can significantly improve the therapeutic activity of anti-ErbB-2 mAb in immunocompetent mice . Expression of genes related to prostaglandin synthesis or signaling in human subcutaneous and omental adipose tissue : depot differences and modulation by adipogenesis . OBJECTIVES : ( 1 ) To examine depot-specific DB00917 and PGF2α release and mRNA expression of enzymes or receptors involved in PG synthesis or signaling in human adipose tissues ; ( 2 ) to identify changes in expression of these transcripts through preadipocyte differentiation ; and ( 3 ) to examine associations between adipose tissue mRNA expression of these transcripts and adiposity measurements . METHODS : Fat samples were obtained surgically in women . DB00917 and PGF2α release by preadipocytes and adipose tissue explants was measured . Expression levels of mRNA coding for enzymes or receptors involved in PG synthesis or signaling were measured by RT-PCR . RESULTS : Cultured preadipocytes and explants from omental fat released more DB00917 and PGF2α than those from the subcutaneous depot and the corresponding transcripts showed consistent depot differences . Following preadipocyte differentiation , expression of P53816 and P43115 mRNA was significantly increased whereas P23219 , P35354 , Q16647 , and O14684 mRNA abundance were decreased in both compartments ( P ≤ 0.01 for all ) . Transcripts that were stimulated during adipogenesis were those that correlated best with adiposity measurements . CONCLUSION : Cells from the omental fat compartment release more DB00917 and PGF2α than those from the subcutaneous depot . Obesity modulates expression of PG-synthesizing enzymes and PG receptors which likely occurs through adipogenesis-induced changes in expression of these transcripts . 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 . 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 . 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 . Circulating apoptotic proteins are increased in long-term disease-free breast cancer survivors . Circulating apoptotic proteins are increased in patients with heart failure . We evaluated whether circulating soluble ( s ) apoptosis-related proteins and inflammation markers are increased in long-term disease free breast cancer survivors and associated with cardiotoxicity , and if subgroups could be identified based on the applied treatments . Circulating tumour necrosis factor ( P01375 ) alpha , sTNF-receptor ( sTNF-R ) 1 and 2 , sFas , sFas ligand , sTNF-related apoptosis inducing ligand ( sTRAIL ) and serum P04626 were measured with immunoassay . High-sensitivity P02741 ( HS-CRP ) , fibrinogen , plasma B-type and N-terminal atrial natriuretic peptide ( NT- P01160 and DB04899 ) were also determined . Thirty-four patients with median 6.0 years follow-up and 12 healthy age-matched women were enrolled . Chemotherapy , consisting of five cycles fluorouracil , epirubicin ( 90 mg/m(2) ) , cyclophosphamide ( FEC ) ( n=14 ) or four cycles FEC followed by myeloablation with high-dose carboplatin , cyclophosphamide , thiotepa ( n=20 ) , preceded irradiation and tamoxifen . Circulating apoptosis markers were higher in patients than in controls . No associations with cardiac dysfunction were observed . sFas ligand and sTRAIL were higher in the high-dose than in the standard-dose group . In conclusion , we observed increased circulating apoptotic protein levels in long-term disease-free breast cancer survivors , treated with adjuvant chemoradiotherapy , particularly after myeloablative chemotherapy . The potential relation with late cardiotoxicity of antineoplastic therapy deserves further study . 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 . delta- DB00855 dehydrase : a new genetic polymorphism in man . A method has been developed for the electrophoretic and quantitative analyses of human red cell delta-aminolevulinate dehydrase ( P13716 ) . The enzyme is under the control of an autosomal gene , with two common codominant alleles . ALADH1 and ALADH2 , with frequencies of 0.89 and 0.11 , respectively , in the Italian population . Mean phenotypic enzyme activities are nearly identical : 52, . 49 and 55 mIU/g Hb for P13716 1 , P35326 and 2 phenotypes respectively . Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells . Antibody-drug conjugates ( ADC ) are emerging as powerful treatment strategies with outstanding target-specificity and high therapeutic activity in patients with cancer . DB08870 represents a first-in-class ADC directed against P28908 (+) malignancies . We hypothesized that its sustained clinical responses could be related to the stimulation of an anticancer immune response . In this study , we demonstrate that the dolastatin family of microtubule inhibitors , from which the cytotoxic component of brentuximab vedotin is derived , comprises potent inducers of phenotypic and functional dendritic cell ( DC ) maturation . In addition to the direct cytotoxic effect on tumor cells , dolastatins efficiently promoted antigen uptake and migration of tumor-resident DCs to the tumor-draining lymph nodes . Exposure of murine and human DCs to dolastatins significantly increased their capacity to prime T cells . Underlining the requirement of an intact host immune system for the full therapeutic benefit of dolastatins , the antitumor effect was far less pronounced in immunocompromised mice . We observed substantial therapeutic synergies when combining dolastatins with tumor antigen-specific vaccination or blockade of the P18621 - Q9NZQ7 and P16410 coinhibitory pathways . Ultimately , treatment with ADCs using dolastatins induces DC homing and activates cellular antitumor immune responses in patients . Our data reveal a novel mechanism of action for dolastatins and provide a strong rationale for clinical treatment regimens combining dolastatin-based therapies , such as brentuximab vedotin , with immune-based therapies . 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 . 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 . Akt activation by arachidonic acid metabolism occurs via oxidation and inactivation of P60484 tumor suppressor . P35354 ( P35354 ) and P09917 ( 5- P28300 ) enzymes are overexpressed during inflammation and multistage tumor progression in many neoplastic disorders including lung , breast and pancreatic cancers . Here we report that the tumor suppressor phosphatase and tensin homolog ( P60484 ) is oxidized and inactivated during arachidonic acid ( AA ) metabolism in pancreatic cancer cell lines expressing P35354 or 5- P28300 . Oxidation of P60484 decreases its phosphatase activity , favoring increased phosphatidylinositol 3,4,5-triphosphate production , activation of Akt and phosphorylation of downstream Akt targets including GSK-3beta and S6K . These effects are recapitulated with pancreatic phospholipase A(2) , which hydrolyses the release of membrane-bound AA . Interference with P60484 's physiological antagonism of signals from growth factors , insulin and oncogenes may confer risk for hypertrophic or neoplastic diseases associated with chronic inflammation or unwarranted oxidative metabolism of essential fatty acids . 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 . 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 .

MH_train_1569

MH_train_1569

MH_train_1569

interacts_with DB08820?

multiple_choice

' A variant of uncertain significance ' and the proliferation of human disease gene databases . The rapid accumulation of mutation data has led to the creation of nearly 300 locus-specific mutation databases . These sites may contain a few dozen to almost 20,000 mutations for a given gene . Many of the mutations are uncharacterised and have no known effects on the gene product , the ' variant of uncertain significance ' . Here , the statistics of mutation distribution are examined for six different gene databases : P38398 and P51587 , haemoglobin-beta ( P68871 ) , P00492 , P13569 and P04637 . The percentage of all possible point mutations for a protein ( the mutation space ) is calculated for each gene and the question ' How much mutation data is enough ? ' is raised . Genetics and treatment options for recurrent acute and chronic pancreatitis . Worldwide research efforts demonstrate a major role of gene-environment interactions for the risk , development , and progression of most pancreatic diseases , including recurrent acute and chronic pancreatitis . New findings of pancreas disease-associated risk variants have been reported in the P15085 , P19440 , P57739 , P03956 , P42898 , and other genes . These risk genes and their regulatory regions must be added to the known pathogenic variants in the P07477 , P00995 , P13569 , Q99895 , P41180 , Q8IWV7 , Q9Y3A5 , P19835 , and P07858 genes . This new knowledge promises to improve disease management and prevention through personalized medicine . At the same time , however , knowledge of an increasing number of pathogenic variants , and their complicated effects when present in combination , results in increasing difficulty in interpretation and development of recommendations . Direct-to-consumer marketing of genetic testing results also adds complexity to disease management paradigms , especially without interpretation and , in many cases , proven accuracy . While improvements in the ability to rapidly and accurately interpret complex genetic tests are clearly needed , some results , such as pathogenic P13569 variants , including a new class of bicarbonate-defective mutations , and P07477 variants have immediate implications that direct management . In addition , discovery of pancreatitis-associated genetic variants in patients with glucose intolerance may suggest underlying type 3c diabetes , which also has implications for treatment and disease management . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . Genetics of idiopathic disseminated bronchiectasis . Bronchiectasis is an abnormal dilation of bronchi , consequent to the destruction of their walls . It is included in the category of obstructive pulmonary diseases , along with chronic obstructive pulmonary disease ( P48444 ) , asthma , and cystic fibrosis . In approximately 50 % of cases , bronchiectasis is associated with underlying conditions ; in the remainder , known causes are not ascertainable ( idiopathic bronchiectasis ) . A search for genetic determinants of this phenotype , with the cystic fibrosis gene as a candidate , has been performed by three independent groups . The results of this search agreed on the association of bronchiectasis with cystic fibrosis gene mutations and polymorphisms . The cystic fibrosis gene is also associated with bronchiectasis due to rheumatoid arthritis and allergic bronchopulmonary aspergillosis . A few other genes have been investigated in idiopathic bronchiectasis , with negative results . Idiopathic bronchiectasis is , therefore , to be considered as an obstructive multifactorial disorder belonging to the category of cystic fibrosis monosymptomatic diseases ( or P13569 -opathies ) , whose pathogenesis is influenced by environmental factors and other undetermined genes . Phosphodiesterase 4 inhibitors for the treatment of asthma and P48444 . Type 4 cyclic nucleotide phosphodiesterases ( PDE4s ) are metallo-hydrolases which specifically hydrolyze DB02527 to AMP in various cells types . The catalytic core is a bimetallic ion center composed of a tightly bound Zn(2+) and a loosely bound Mg(2+) , which plays a dictating role in eliciting DB02527 binding and catalysis activation . An invariant glutamine positioned opposite to the ion center serves as the substrate recognition determinant and synergizes the transient Mg-oxo-phosphate interaction in the substrate complex . The Mg(2+) binding is activated by a PKA-mediated serine phosphorylation and modulated through protein-protein interactions , thus , providing efficient mechanisms in the temporal regulation of DB02527 signaling . Several DB05876 inhibitors including roflumilast , cilomilast and rolipram also rely on the interaction with the glutamine and metallic ion center for binding , with their affinity enhanced dramatically by the presence of the Mg(2+) ion . Recent studies have provided new insights into the role of this enzyme in inflammatory settings , P13569 regulation , long term potentiation , and its importance in immune surveillance . The major inflammatory cytokines which are modulated with DB05876 inhibitors include TNFalpha , P60568 , IFNgamma , IL-12 , GM- P04141 and Q06643 (4) . The role of DB05876 inhibitors in modulating cytokines , lipid mediators and in mucociliary clearance , along with clinical efficacy in asthma and/or P48444 demonstrated with roflumilast and cilomilast , suggest a broad anti-inflammatory spectrum for these compounds . Presently , the major impediment to approval of these novel therapies has been the mechanism based gastrointestinal adverse events which has limited the dosing and the ultimate efficacy with these novel therapeutic agents . Development , clinical utility , and place of ivacaftor in the treatment of cystic fibrosis . Cystic fibrosis ( CF ) is a life-limiting , multisystem disease characterized by thick viscous secretions leading to recurrent lung infections , bronchiectasis , and progressive deterioration in lung function . CF is caused by loss or dysfunction of the CF transmembrane conductance regulator ( P13569 ) protein which is responsible for transepithelial chloride and water transport . Improved understanding of P13569 protein dysfunction has allowed the development of mutation-specific small-molecule compounds which directly target the underlying P13569 defect . DB08820 is the first licensed small-molecule compound for CF patients which targets the P13569 gating mutation Gly551Asp ( previously termed G551D ) and has the potential to be truly disease-modifying . DB08820 is an oral medication given twice daily and has shown benefit in terms of an increase in lung function , decreased sweat chloride , weight gain , improvement in patient-reported quality of life , and reduction in number of respiratory exacerbations in clinical trials . Although ivacaftor is currently only licensed for use in approximately 5 % of the CF population ( those who have at least one Gly551Asp mutation ) , the developmental pathway established by ivacaftor paves the way for other P13569 modulators that may benefit many more patients . In particular , a P13569 modulator for those with the Phe508del deletion ( previously ∆F508 ) would allow 90 % of the CF population to benefit from disease-modifying treatment . 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 . 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 . Novel role of cystic fibrosis transmembrane conductance regulator in maintaining adult mouse olfactory neuronal homeostasis . The olfactory epithelium ( OE ) of mice deficient in cystic fibrosis transmembrane conductance regulator ( P13569 ) exhibits ion transport deficiencies reported in human CF airways , as well as progressive neuronal loss , suggesting defects in olfactory neuron homeostasis . Microvillar cells , a specialized OE cell-subtype , have been implicated in maintaining tissue homeostasis . These cells are endowed with a PLCβ2/IP3 R3/ Q9Y210 signal transduction pathway modulating release of neuropeptide Y ( P01303 ) , which stimulates OE stem cell activity . It is unknown , however , whether microvillar cells also mediate the deficits observed in P13569 -null mice . Here we show that Cftr mRNA in mouse OE is exclusively localized in microvillar cells and P13569 immunofluorescence is coassociated with the scaffolding protein O14745 and PLCβ2 in microvilli . In P13569 -null mice , PLCβ2 was undetectable , O14745 mislocalized , and IP3 R3 more intensely stained , along with increased levels of P01303 , suggesting profound alteration of the PLCβ2/IP3 R3 signaling pathway . In addition , basal olfactory neuron homeostasis was altered , shown by increased progenitor cell proliferation , differentiation , and apoptosis and by reduced regenerative capacity following methimazole-induced neurodegeneration . The importance of P13569 in microvillar cells was further underscored by decreased thickness of the OE mucus layer and increased numbers of immune cells within this tissue in P13569 -KO mice . Finally , we observed enhanced immune responses to an acute viral-like infection , as well as hyper-responsiveness to chemical and physical stimuli applied intranasally . Taken together , these data strengthen the notion that microvillar cells in the OE play a key role in maintaining tissue homeostasis and identify several mechanisms underlying this regulation through the multiple functions of P13569 . P25025 -dependent mucosal neutrophil influx protects against colitis-associated diarrhea caused by an attaching/effacing lesion-forming bacterial pathogen . Enteropathogenic Escherichia coli ( EPEC ) is a major cause of diarrheal disease in young children , yet symptoms and duration are highly variable for unknown reasons . Citrobacter rodentium , a murine model pathogen that shares important functional features with EPEC , colonizes mice in colon and cecum and causes inflammation , but typically little or no diarrhea . We conducted genome-wide microarray studies to define mechanisms of host defense and disease in C. rodentium infection . A significant fraction of the genes most highly induced in the colon by infection encoded CXC chemokines , particularly P09341 /2/5 and Q07325 /10 , which are ligands for the chemokine receptors P25025 and P49682 , respectively . CD11b(+) dendritic cells were the major producers of P09341 , P42830 , and Q07325 , while P19875 was mainly induced in macrophages . Infection of gene-targeted mice revealed that P49682 had a significant but modest role in defense against C. rodentium , whereas P25025 had a major and indispensable function . P25025 was required for normal mucosal influx of neutrophils , which act as direct antibacterial effectors . Moreover , P25025 loss led to severe diarrhea and failure to express critical components of normal ion and fluid transport , including ATPase beta(2)-subunit , P13569 , and P40879 . The antidiarrheal functions were unique to P25025 , since other immune defects leading to increased bacterial load and inflammation did not cause diarrhea . Thus , P25025 -dependent processes , particularly mucosal neutrophil influx , not only contribute to host defense against C. rodentium , but provide protection against infection-associated diarrhea . Specific expression of matrix metalloproteinases 1 , 3 , 9 and 13 associated with invasiveness of breast cancer cells in vitro . Several matrix metalloproteinases ( MMPs ) and tissue inhibitors of MMPs ( TIMPs ) were studied in highly invasive ( MDA-MB-231 ) and slightly invasive ( MCF-7 , T47D , BT-20 ) breast cancer cell lines . Investigations were carried out at the protein level and/or at the mRNA level , either in cells cultured as monolayers on plastic , or in cells seeded on a thin layer of Matrigel basement membrane matrix . Analysis of MMP expression by RT-PCR showed expression of P03956 . P08254 , and P45452 in highly invasive MDA-MB-231 cells , but not in slightly invasive cell lines . The extracellular secretion of P03956 and P08254 by MDA-MB 231 cells could be also shown by ELISA . P01033 and P16035 mRNAs were found in all cell lines , however , the extracellular secretion of both TIMPs was much higher in MDA-MB-231 cells than in the other cell lines . When the cells were cultured on Matrigel matrix , P14780 expression was induced in MDA-MB-231 cells only , as assessed by RT-PCR and zymography experiments . The invasive potential of MDA-MB-231 cells evaluated in vitro through Matrigel was significantly inhibited by the MMP inhibitor DB00786 , by 25 % and 50 % at the concentrations of 2 x 10(-6) M and 10(-5) M , respectively . In conclusion , our data show that highly invasive MDA-MB-231 cells but not slightly invasive T47D , MCF-7 and BT-20 cells express P03956 , P08254 , P14780 and P45452 . P14780 which is specifically up-regulated by cell contact to Matrigel , may play a key role in the invasiveness of MDA-MB-231 cells through basement membranes . [ 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 . P10997 -driven metabolic reprogramming induces regression of p53-deficient tumours in vivo . P04637 is commonly altered in human cancer , and Tp53 reactivation suppresses tumours in vivo in mice ( P04637 and Tp53 are also known as p53 ) . This strategy has proven difficult to implement therapeutically , and here we examine an alternative strategy by manipulating the p53 family members , Tp63 and Tp73 ( also known as p63 and p73 , respectively ) . The acidic transactivation-domain-bearing ( TA ) isoforms of p63 and p73 structurally and functionally resemble p53 , whereas the ΔN isoforms ( lacking the acidic transactivation domain ) of p63 and p73 are frequently overexpressed in cancer and act primarily in a dominant-negative fashion against p53 , TAp63 and TAp73 to inhibit their tumour-suppressive functions . The p53 family interacts extensively in cellular processes that promote tumour suppression , such as apoptosis and autophagy , thus a clear understanding of this interplay in cancer is needed to treat tumours with alterations in the p53 pathway . Here we show that deletion of the ΔN isoforms of p63 or p73 leads to metabolic reprogramming and regression of p53-deficient tumours through upregulation of P10997 , the gene that encodes amylin , a 37-amino-acid peptide co-secreted with insulin by the β cells of the pancreas . We found that P10997 is causally involved in this tumour regression and that amylin functions through the calcitonin receptor ( CalcR ) and receptor activity modifying protein 3 ( O60896 ) to inhibit glycolysis and induce reactive oxygen species and apoptosis . DB01278 , a synthetic analogue of amylin that is currently used to treat type 1 and type 2 diabetes , caused rapid tumour regression in p53-deficient thymic lymphomas , representing a novel strategy to target p53-deficient cancers . DB06698 ameliorates olanzapine-induced weight gain through modulation of histaminergic , P01303 and AMPK pathways . Olanzapine is widely used to treat schizophrenia and other disorders , but causes adverse obesity and other metabolic side-effects . Both animal and clinical studies have shown that co-treatment with betahistine ( a histaminergic H1 receptor agonist and H3 receptor antagonist ) is effective for ameliorating olanzapine-induced weight gain/obesity . To reveal the mechanisms underlying these effects , this study investigated the effects of co-treatment of olanzapine and betahistine ( O+B ) on expressions of histaminergic H1 receptor ( P35367 ) , AMP-activated protein kinase ( AMPK ) , neuropeptide Y ( P01303 ) , and proopiomelanocortin ( P01189 ) in the hypothalamus associated with reducing olanzapine-induced weight gain . Olanzapine significantly upregulated the mRNA and protein expressions of P35367 , while O+B co-treatment significantly downregulated the P35367 levels , compared to the olanzapine-only treatment group . The P01303 mRNA expression was significantly enhanced by olanzapine , but it was significantly reversed by O+B co-treatment . The hypothalamic P35367 expression was positively correlated with total food intake , and P01303 expression . Olanzapine also increased AMPKα activation measured by the AMPKα phosphorylation ( pAMPKα ) /AMPKα ratio compared with controls , whereas O+B co-treatment decreased the pAMPKα/AMPKα ratio , compared with olanzapine only treatment . The pAMPKα/AMPKα ratio was positively correlated with total food intake and P35367 expression . Although olanzapine administration decreased the P01189 mRNA level , this level was not affected by O+B co-treatment . Therefore , these results suggested that co-treatment with betahistine may reverse olanzapine-induced body weight gain via the P35367 - P01303 and P35367 -pAMPKα pathways . Novel 3-substituted rimonabant analogues lack Δ(9) -tetrahydrocannabinol-like abuse-related behavioural effects in mice . BACKGROUND AND PURPOSE : Previous structure-activity relationship studies with analogues of the P21554 receptor antagonist rimonabant have demonstrated that a subset of these analogues with 3-substituent replacements of rimonabant 's pyrazole core displayed cannabimimetic profiles seemingly independent of P21554 receptors . We sought to further evaluate these analogues in several behavioural models sensitive to detecting THC-like abuse liability . EXPERIMENTAL APPROACH : Selected analogues were tested in a battery of tests in mice to replicate previous findings . Cross-generalization tests were conducted in mice trained to discriminate either THC or O-6629 from vehicle . DB06155 and its analogues were also evaluated in substitution and challenge tests . Finally , development of cross-tolerance between THC and O-6211 in the mouse test battery was assessed . KEY RESULTS : O-6629 and O-6658 produced dose-dependent acute cannabimimetic activity in mice , but neither substituted for nor antagonized THC 's discriminative stimulus . Cross-substitution was observed with O-6658 in mice discriminating O-6629 , whereas rimonabant neither substituted for nor attenuated the O-6629 discriminative stimulus . THC and morphine did not generate O-6629-like responding . Cross-tolerance did not develop in mice repeatedly treated with THC when tested with O-6211 in the mouse test battery . CONCLUSIONS AND IMPLICATIONS : While some overlap exists between the pharmacological profiles of THC and these 3-substituent rimonabant analogues , the effects are mediated by distinct neural targets . Notably , these analogues are unlikely to possess marijuana-like abuse liability in humans , but general abuse liability has not yet been determined . Efforts to determine the mechanism(s) of action of this seemingly unique class of compounds are underway . P13569 modulates neurosecretory function in pulmonary neuroendocrine cell-related tumor cell line models . The pulmonary neuroendocrine cell ( PNEC ) system consists of solitary cells and distinctive cell clusters termed neuroepithelial bodies ( P20929 ) localized in the airway epithelium . PNEC/ P20929 express a variety of bioactive substances , including amine ( serotonin , 5HT ) and neuropeptides . We have previously shown that P20929 cells are O(2) sensors expressing nicotinamide adenine diphosphate oxidase complex and O(2) sensitive K(+) channel . Recently , we demonstrated expression of functional cystic fibrosis transmembrane conductance regulator ( P13569 ) and Cl(-) conductances in P20929 cells of rabbit neonatal lung . Because PNEC/ P20929 are sparsely distributed and difficult to study in native lung , we investigated small-cell lung carcinoma ( SCLC ) and carcinoid tumor cell lines ( tumor counterparts of normal PNEC/ P20929 ) as models for PNEC/ P20929 . SCLC ( H146 , H345 ) and carcinoid ( H727 ) cell lines express neuroendocrine cell markers , including chromogranin A , neural cell adhesion molecule ( N- P62158 ) , 5HT , and tryptophan hydroxylase . We report that H146 , H345 , and H727 express P13569 messenger RNA ( reverse transcription polymerase chain reaction ) and protein ( immunoblotting ) and possess functional P13569 Cl(-) conductance , demonstrated by an iodide efflux assay inhibitable by transfection with antisense P13569 . Using an immunoassay to quantitate 5HT secretion , we also show that downregulation of P13569 abolishes hypoxia-induced 5HT release , and reduces secretory response to high potassium . Our findings suggest that P13569 may modulate neurosecretory activity of PNEC/ P20929 possessing O(2) sensor function . We propose that these tumor cell lines may be useful models for investigating the role of P13569 in PNEC/ P20929 functions in health and 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 . NDPK-A ( but not NDPK-B ) and AMPK alpha1 ( but not AMPK alpha2 ) bind the cystic fibrosis transmembrane conductance regulator in epithelial cell membranes . Cystic fibrosis ( CF ) results from mutations within the cystic fibrosis transmembrane-conductance regulator ( P13569 ) protein . The AMP-activated protein kinase ( AMPK ) is a heterotrimer composed of different isoforms of the alphabetagamma subunits , where the alpha1 catalytic subunit binds P13569 . Nucleoside diphosphate kinase ( NDPK , P15531 /awd ) converts nucleoside diphosphates to nucleoside triphosphates but also acts as a protein kinase . We recently showed that AMPK alpha1 binds NDPK-A in lung epithelial cytosol . Here we report that in the plasma membrane of human airway epithelial cells , NDPK-A and AMPK alpha1 associate with the plasma membrane via P13569 . We show that the regulatory domain of P13569 binds NDPK-A whereas P54619 or gamma2 bind the first nucleotide binding domain ( NBD1 ) and AMPK alpha1 binds the second ( NBD2 ) of P13569 . We also show that NDPK-A specifically binds AMPK alpha1 and Q9UGJ0 subunits , thereby specifying the isozyme of AMPK heterotrimer that associates with P13569 at the membrane . Thus , the combined data provide novel insight into the subunit composition of the epithelial P13569 /AMPK/NDPK complex , such that : P13569 interacts specifically with AMPK alpha1 , gamma2 and NDPK-A and not NDPK-B or P54619 . Dual inhibitors of cyclooxygenase and P09917 . A new avenue in anti-inflammatory therapy ? Nonsteroidal anti-inflammatory drugs ( NSAIDs ) are a mainstay in the treatment of inflammatory disease and are among the most widely used drugs worldwide . They are anti-inflammatory , antipyretic , and analgesic and are prescribed as first choice for the treatment of rheumatic disorders and , in general , inflammation . The main limitation in using NSAIDs consists in their side-effects , including gastrointestinal ulcerogenic activity and bronchospasm . The mechanism of action of these drugs is attributed to the inhibition of cyclooxygenase ( P36551 ) , and , consequently , the conversion of arachidonic acid into prostaglandins . It is hypothesized that the undesirable side-effects of NSAIDs are due to the inhibition of P23219 ( constitutive isoform ) , whereas the beneficial effects are related to the inhibition of P35354 ( inducible isoform ) . Arachidonic acid can also be converted to leukotrienes ( LTs ) by the action of P09917 ( 5- P28300 ) . LTC ( 4 , ) LTD ( 4 , ) and LTE(4) are potent bronchoconstrictors , whereas Q06643 (4) is chemotactic for leukocytes and plays an important role in the development of gastrointestinal ulcers by contributing to the inflammatory process . Thus , developing dual inhibitor compounds that will simultaneously inhibit P36551 and 5- P28300 could enhance their individual anti-inflammatory effects and reduce the undesirable side-effects associated with NSAIDs , especially of the gastrointestinal tract . The most promising P36551 /5- P28300 inhibitor is DB04725 ( [ 2,2-dimethyl-6-(4-chlorophenyl)-7-phenyl-2,3-dihydro-1H-pyrrolizine-5-yl ] -acetic acid ) , now in Phase III clinical trials . This new approach will certainly help to unravel the mechanisms at the root of the undesirable effects of NSAIDs and to develop safer NSAIDs . 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 . The elmiric acids : biologically active anandamide analogs . As chemical entities , lipoamino acids have been known for some time . However , more recently their occurrence and importance in mammalian species has been discovered . They appear to have close relationships with the endocannabinoids not only structurally but also in terms of biological actions . The latter include analgesia , anti-inflammatory effects , inhibition of cell proliferation and calcium ion mobilization . To date about 40 naturally occurring members of this family have been identified and , additionally , several synthetic analogs have been prepared and studied . To facilitate their identity , a nomenclature system has been suggested based on the name elmiric acid ( P15941 ) . The prototypic example , N-arachidonoyl glycine , does not bind to P21554 , however it does inhibit the glycine transporter GLYT2a and also appears to be a ligand for the orphan G-protein-coupled receptor Q14330 . It may also have a role in regulating tissue levels of anandamide by virtue of its inhibitory effect on FAAH the enzyme that mediates inactivation of anandamide . Its concentration in rat brain is several-fold higher than anandamide supporting its possible role as a physiological mediator . Future studies should be aimed at elucidating the actions of all of the members of this interesting family of molecules .

MH_train_1570

MH_train_1570

MH_train_1570

interacts_with DB06684?

multiple_choice

Obestatin stimulates Akt signalling in gastric cancer cells through beta-arrestin-mediated epidermal growth factor receptor transactivation . Obestatin was identified as a gut peptide encoded by the ghrelin gene that interacts with the G protein-coupled receptor , O43194 . In this work , a sequential analysis of its transmembrane signalling pathway has been undertaken to characterize the intracellular mechanisms responsible for Akt activation . The results show that Akt activation requires the phosphorylation of T308 in the A-loop by the phosphoinositide-dependent kinase 1 ( PDK1 ) and S473 within the HM by the mammalian target of rapamycin ( P42345 ) kinase complex 2 ( mTORC2 : Rictor , Q9BVC4 , mSin1 , P42345 kinase ) with participation neither of G(i)(/o)-protein nor Gbetagamma dimers . Obestatin induces the association of O43194 /beta-arrestin 1/Src signalling complex resulting in the transactivation of the epidermal growth factor receptor ( P00533 ) and downstream Akt signalling . Upon administration of obestatin , phosphorylation of P42345 ( S2448 ) and p70S6K1 ( T389 ) rise with a time course that parallels that of Akt activation . Based on the experimental data obtained , a signalling pathway involving a beta-arrestin 1 scaffolding complex and P00533 to activate Akt signalling is proposed . Anti-vascular endothelial growth factor therapy for ocular neovascular disease . PURPOSE OF REVIEW : Recent research has shown that vascular endothelial growth factor ( P15692 ) is responsible for many ocular pathologies involving neovascularization . Over the past several years several new agents targeting P15692 have become commercially available for intraocular use . These agents have revolutionized the care of neovascular age related macular degeneration and have great potential for other blinding conditions such as diabetic retinopathy , retinopathy of prematurity , and neovascular glaucoma . RECENT FINDINGS : The P15692 Inhibition Study in Ocular Neovascularization ( VISION ) trial first showed that an anti- P15692 agent ( pegaptanib ) was able to prevent vision loss in neovascular age related macular degeneration . The Minimally Classic/Occult Trial of Anti- P15692 Antibody DB01270 in the Treatment of Neovascular AMD ( MARINA ) and Anti- P15692 Antibody for the Treatment of Predominantly Classic Choroidal Neovascularization in AMD ( ANCHOR ) trials showed that ranibizumab prevented moderate vision loss in neovascular age related macular degeneration and for the first time that a substantial proportion of patients regained vision . Smaller case series have shown that bevacizumab can regress retinal , iris and disc neovascularization . Ongoing trials are investigating the utility of anti- P15692 therapy in retinopathy of prematurity , diabetic retinopathy , and neovascular glaucoma . SUMMARY : Newer anti- P15692 therapies have shown unprecedented efficacy in treating age related macular degeneration with many patients experiencing improvement in vision . Ongoing trials will help guide their use in age related macular degeneration and expand their indications to many other blinding diseases . Rindopepimut , a 14-mer injectable peptide vaccine against EGFRvIII for the potential treatment of glioblastoma multiforme . Celldex Therapeutics is developing rindopepimut ( DB05374 ) , a 14-mer injectable peptide vaccine for the potential treatment of glioblastoma multiforme ( GBM ) . Rindopepimut specifically targets a novel junctional epitope of the P00533 deletion mutant EGFRvIII , which is a constitutively active receptor that is expressed in approximately 60 to 70 % of patients with GBM . EGFRvIII expression is correlated with worse prognosis and reduced overall survival . Importantly , EGFRvIII is not expressed in normal brain tissue , making it an excellent therapeutic target . Preclinical studies demonstrated lasting tumor regression and increased survival times , as well as efficient generation of EGFRvIII-specific humoral and cellular immune responses , in animals expressing EGFRvIII and vaccinated with rindopepimut . Phase I and II clinical trials in patients with GBM demonstrated significantly increased median time to progression and overall survival time in those vaccinated with rindopepimut compared with matched historical controls . Only limited side effects have been observed in patients . Given these results , rindopepimut is an extremely promising therapy for patients with GBM . Phase I and II clinical trials in patients with GBM were ongoing at the time of publication . In the future , it may be beneficial to combine rindopepimut with other treatment modalities to further prolong survival . 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 . 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 . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . DB06016 , an orally active D2/D3 receptor antagonist , for the potential treatment of schizophrenia , bipolar mania and depression . DB06016 ( RGH-188 ) , which is being codeveloped by Gedeon Richter Ltd , Forest Laboratories Inc and Mitsubishi Tanabe Pharma Corp , is a novel putative antipsychotic drug that exerts partial agonism at dopamine D2/D3 receptors , with preferential binding to D3 receptors , and partial agonism at serotonin P08908 receptors . Its activity at D2/D3 receptors may be lower than that of the prototype partial agonist aripiprazole . The antipsychotic activity of cariprazine was demonstrated in animal models , and data also suggest that the propensity for extrapyramidal side effects is low and that the drug may have procognitive properties . DB06016 is rapidly absorbed , with high oral bioavailability and a long plasma elimination t1/2 . DB06016 is in phase III clinical trials in patients with schizophrenia and in patients with bipolar disorder . Data from phase II trials in patients with schizophrenia and bipolar mania indicate that the drug has antipsychotic and antimanic properties that are superior to placebo . With its unique receptor affinity profile , cariprazine may represent a potential enrichment of the therapeutic armamentarium for schizophrenia and affective disorders . Its activity against the cognitive deficits associated with schizophrenia has to be carefully investigated . 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 . 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 . Changes of several brain receptor complexes in the cerebral cortex of patients with Alzheimer disease : probable new potential pharmaceutical targets . Although Alzheimer disease ( AD ) has been linked to defects in major brain receptors , studies thus far have been limited to the determination of receptor subunits or specific ligand binding studies . However , the availability of current technology enables the determination and quantification of brain receptor complexes . Thus , we examined levels of native receptor complexes in the brains of patients with AD . Cortical tissue was obtained from control subjects ( n = 12 females and 12 males ) and patients with AD ( n = 12 females and 12 males ) within a 3-h postmortem time period . The tissues were kept frozen until further biochemical analyses . Membrane proteins were extracted and subsequently enriched by ultracentrifugation using a sucrose gradient . Membrane proteins were then electrophoresed onto native gels and immunoblotted using antibodies against individual brain receptors . We found that the levels were comparable for complexes containing GluR2 , GluR3 and P48058 as well as P08908 . Moreover , the levels of complexes containing muscarinic AChR M1 , Q9UHB4 and GluR1 were significantly increased in male patients with AD . Nicotinic AChRs 4 and 7 as well as dopaminergic receptors D1 and D2 were also increased in males and females with AD . These findings reveal a pattern of altered receptor complex levels that may contribute to the deterioration of the concerted activity of these receptors and thus result in cognitive deficits observed in patients with AD . It should be emphasised that receptor complexes function as working units rather than individual subunits . Thus , the receptor deficits identified may be relevant for the design of experimental therapies . Therefore , specific pharmacological modulation of these receptors is within the pharmaceutical repertoire . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . 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 . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . [ Leukemia- and lymphoma-associated flow cytometric , cytogenetic , and molecular genetic aberrations in healthy individuals ] . Most leukemia and lymphoma cases are characterized by specific flow cytometric , cytogenetic and molecular genetic aberrations , which can also be detected in healthy individuals in some cases . The authors review the literature concerning monoclonal B-cell lymphocytosis , and the occurrence of chromosomal translocations t(14;18) and t(11;14) , P06748 - Q9UM73 fusion gene , O60674 V617F mutation , P11274 - P00519 fusion gene , P41212 - Q01196 ( P41212 - Q01196 ) , Q03164 - P51825 and P29590 - P10276 fusion gene in healthy individuals . At present , we do not know the importance of these aberrations . From the authors review it is evident that this phenomenon has both theoretical and practical ( diagnostic , prognostic , and therapeutic ) significance . 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 . 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 . Impact of wines and wine constituents on cyclooxygenase-1 , cyclooxygenase-2 , and P09917 catalytic activity . Cyclooxygenases and lipoxygenases are proinflammatory enzymes ; the former affects platelet aggregation , vasoconstriction , vasodilatation and later the development of atherosclerosis . Red wines from Georgia and central and western Europe inhibited cyclooxygenase-1 ( P23219 ) activity in the range of 63-94 % , cyclooxygenase-2 ( P35354 ) activity in the range of 20-44 % ( tested at a concentration of 5 mL/L ) , and P09917 ( 5- P28300 ) activity in the range of 72-84 % ( at a concentration of 18.87 mL/L ) . White wines inhibited 5- P28300 in the range of 41-68 % at a concentration of 18.87 mL/L and did not inhibit P23219 and P35354 . Piceatannol ( IC50 = 0.76 μM ) was identified as a strong inhibitor of 5- P28300 followed by luteolin ( IC50 = 2.25 μM ) , quercetin ( IC50 = 3.29 μM ) , and myricetin ( IC50 = 4.02 μM ) . trans- DB02709 was identified as an inhibitor of P23219 ( IC50 = 2.27 μM ) and P35354 ( IC50 = 3.40 μM ) . Red wine as a complex mixture is a powerful inhibitor of P23219 , P35354 , and 5- P28300 , the enzymes involved in eicosanoid biosynthetic pathway . 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 . 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 . 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 . P02787 and ferritin modulate the activity of brain calcium-calmodulin-dependent phosphodiesterase . The effect of the key iron homeostasis proteins transferrin and ferritin on the activity of partially purified brain calcium-calmodulin-dependent phosphodiesterase ( P62158 -PDE , EC 3.4.1.17 ) were studied . P02787 and ferritin were found to be potent natural activators of P62158 -PDE . The key factor determining the degree of activation by these proteins is their saturation with iron : apotransferrin activated P62158 -PDE 6-7-fold ; iron-poor brain ferritin and liver apoferritin ( taken for comparison ) activated the enzyme 4-5- and 2-fold , respectively . Diferric transferrin and iron-rich liver ferritin had no effects on the enzyme activity . P02787 and ferritin ( both in apo- and iron-saturated forms ) did not change the activity of calmodulin-phosphodiesterase complex . The data suggest that apotransferrin and iron-poor transferrin are involved in the regulation of cyclic nucleotide content in nervous tissue .

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interacts_with DB00215?

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Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms . A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells , and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase , is described . DB00120 hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity . The phosphorylation site(s) appear to be the same for both kinases . Phosphorylation is accompanied by increased metabolic flux at low , physiologically relevant , substrate concentrations . P01308 and spermine both inhibit the phosphorylation of the enzyme , possibly by increasing dephosphorylation . P17735 is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium . No parallel changes in flux could be detected . Both enzymes are subject to complex regulatory mechanisms , short- and long-term . Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes . Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux . 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 . 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 . 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 ) Low and high levels of DB00163 exert opposite effects on P60568 possibly through the modulation of P37231 , P25963 , and apoptotic pathway in activated splenocytes . OBJECTIVE : We previously demonstrated that a high dose of alpha-tocopheryl succinate inhibits interleukin-2 ( P60568 ) mRNA and production in autoimmune-prone MRL/lpr mice . In the present study , we investigated the regulation of DB00163 ( alphaTOC ) on P60568 gene expression by examining the mRNA of P60568 , inhibitor kappaBalpha ( P25963 ) , and peroxisome proliferator-activated receptor-gamma ( PPARgamma ) . METHODS : Messenger RNA expression in active splenocytes of BALB/c mice was investigated with reverse transcriptase polymerase chain reaction . RESULTS : Levels of P60568 mRNA in phorbol 12-myristate 13-acetate/ionomycin activated splenocytes and cytokine in T-helper-1 cells were increased by 50 microM of alphaTOC but decreased by 1 mM of alphaTOC . In addition , the P25963 gene expression significantly increased by the high dose ( > or=500 microM ) of alphaTOC , suggesting an inhibition on nuclear factor-kappaB pathway for activation of P60568 expression . PPARgamma mRNA level in activated splenocytes was upregulated by 1 mM of alphaTOC . PPARgamma mRNA level in unstimulated splenocytes was upregulated by alphaTOC in a dose-dependent manner , suggesting that alphaTOC might enhance the PPARgamma signaling pathway . High-dose alphaTOC induced apoptosis of splenocytes and inhibited phytohemagglutinin-stimulated T-cell proliferation . Conversely , the proliferative response of splenocytes was enhanced by 5 microM of alphaTOC . Low-dose ( 50 microM ) alphaTOC increased P60568 expression , which may have been due to the absence of downregulation of PPARgamma and P25963 on the P60568 gene . CONCLUSION : The results indicated that low and high doses of alphaTOC exert opposite effects on P60568 , possibly through modulation of PPARgamma , P25963 , and apoptosis pathways . The present findings support our previous observation of opposite effects of low- and high-dose vitamin E on survival of MRL/lpr mice . 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 . Characterization of haloperidol and trifluperidol as subtype-selective N-methyl-D-aspartate ( DB01221 ) receptor antagonists using [3H] DB01520 and [3H]ifenprodil binding in rat brain membranes . [3H] DB01520 and [3H]ifenprodil binding to N-methyl-D-aspartate ( DB01221 ) receptors in rat forebrain membranes was used to compare the inhibition of haloperidol and trifluperidol with that of ifenprodil and eliprodil . In the [3H] DB01520 binding assay , inhibition curves of ifenprodil , eliprodil , haloperidol and trifluperidol revealed two affinity states in the presence of glutamate , glycine and spermidine . The potency of these agents to inhibit the high-affinity fraction of the binding agreed with the results of other studies investigating their potency to block glutamate-induced current at recombinant NR1a/ Q13224 DB01221 receptors expressed in Xenopus oocytes . These agents also inhibited [3H]ifenprodil binding in a biphasic manner , whether in the absence or the presence of either the sigma site ligand GBR-12909 or spermidine . DB03566 reduced the fraction of high-affinity sites labeled with [3H]ifenprodil . The only alteration in the affinity was a decrease in the IC50 value of haloperidol to inhibit the high-affinity fraction of [3H]ifenprodil binding . GBR-12909 also reduced the fraction of [3H]ifenprodil sites inhibited by these compounds with high affinity , with no change in the affinity for either fraction . These data suggest that spermidine is neither a competitive antagonist at the fraction of the binding inhibited by these agents with high affinity , nor is this fraction of the binding to sigma sites . DB00502 and trifluperidol represent a new class of agent that interacts at a site that is labeled by [3H]ifenprodil as well as [3H] DB01520 in rat brain membranes and that closely reflects ifenprodil 's voltage-independent site on the recombinant NR1a/ Q13224 subtype of the DB01221 receptor . Increased P35225 -producing T cells in P35858 : positive correlations with disease severity and progression rate . We measured the intracellular productions of IFNgamma , P60568 , P05112 , P35225 and TNFalpha in peripheral blood P01730 (+) and CD8(+) T cells from 21 amyotrophic lateral sclerosis ( P35858 ) patients , 14 disease controls ( DC ) with spinocerebellar degeneration and 16 healthy controls ( HC ) . Only the percentages of P01730 (+) P35225 (+) and CD8(+) P35225 (+) T cells were significantly higher in P35858 patients than in DC and HC . The P01730 (+) P35225 (+) T cell percentages showed a significant negative correlation with the revised P35858 functional rating scale scores and significant positive correlation with the disease progression rate , suggesting that P35225 contributes to P35858 . Diabetes mellitus in cancer patients treated with combination interleukin 2 and alpha-interferon . Diabetes mellitus is thought to be an autoimmune disease caused by destruction of beta cells in pancreatic islets . P01308 resistance in the peripheral tissues may also play a role . Both interleukin 2 ( P60568 ) and alpha interferon can enhance immune function by stimulating formation of cytolytic T cells and/or antigen expression on both normal and tumor cells . This report describes three patients with advanced malignancy who were treated with combination P60568 and alpha interferon who had the onset or worsening of diabetes mellitus . One patient died as a result . There is evidence that interferon can increase insulin resistance and it is likely that both agents can initiate or enhance an ongoing autoimmune process . Physicians using this combination of drugs should be aware of this potential serious toxicity . Thiol-sensitive mast cell lines derived from mouse bone marrow respond to a mast cell growth-enhancing activity different from both P08700 and P05112 . A series of permanent P08700 -dependent cell lines have been established from normal BALB/c or C3H bone marrow using alpha-thioglycerol-supplemented culture medium and PWM-stimulated spleen cell-conditioned medium as a source of P08700 . The cell lines and derivatives cloned in agar resembled " mucosal type " mast cells with respect to phenotypic and functional properties . In this report we demonstrate that in vitro growth of these mast cell lines was not only dependent on P08700 and synergistically enhanced by P05112 , but in addition regulated by alpha-thioglycerol which could be replaced by 2-ME or cysteamine . We show that these thiol-sensitive mast cell lines respond to a mast cell growth enhancing activity ( MEA ) present in spleen cell-conditioned medium and acting in concert with P08700 . Partially purified MEA was not able to stimulate the growth of P08700 -dependent 32Dcl.23 cells , P60568 -dependent CTLL-2 cells or the mouse T cell line F4/4K.6 ( L3T4+ ) adapted to grow in purified P05112 . Moreover , 11B11 hybridoma-derived anti- P05112 mAb specifically neutralizing mouse Il-4 were unable to abolish the bioactivity of MEA . PWM , P09603 , GM- P04141 , IL-1 , P60568 , P05113 , P05231 , P13232 , P01579 , TGF-alpha , P01375 , P01138 , or EPO did not substitute for MEA in our standard proliferation assay . DB00197 enhances tamoxifen-induced growth inhibitory activity of MCF-7 cells . P37231 ( PPARgamma ) ligands have been identified as a potential source of therapy for human cancers . However , PPARgamma ligands have a limitation for breast cancer therapy , since estrogen receptor alpha ( ER(alpha) ) negatively interferes with PPARgamma signaling in breast cancer cells . Here we show that ER(alpha) inhihits PPARgamma transactivity and ER(alpha)-mediated inhibition of PPARgamma transactivity is blocked by tamoxifen , an estrogen receptor blocker . The activation of ER(alpha) with 17-beta-estradiol blocked PPRE transactivity induced by troglitazone , a PPARgamma ligand , indicating the resistance of ER(alpha)-positive breast cancer cells to troglitazone . Indeed , troglitazone inhibited the growth of ER(alpha)-negative MDA-MB-231 cells more than that of ER(alpha)-positive MCF-7 cells . Combination of troglitazone with tamoxifen led to a marked increase in growth inhibition of ER(alpha)-positive MCF-7 cells compared to either agent alone . Our data indicates that troglitazone enhances the growth inhibitory activity of tamoxifen in ER(alpha)-positive MCF-7 cells . 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 . 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 . 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 . P60568 inhibits DB01221 receptor-mediated currents directly and may differentially affect subtypes . Using whole-cell patch-clamp recordings , this study investigated the effects of interleukin-2 ( P60568 ) on N-methyl-d-aspartate ( DB01221 ) receptor-mediated currents ( I( DB01221 ) ) in rat cultured hippocampal neurons and human embryonic kidney ( P29320 ) 293 cells expressing recombinant DB01221 receptors . We found that P60568 ( 0.01-1ng/ml ) immediately and significantly decreased peak I( DB01221 ) in cultured neurons . Interestingly , the peak I( DB01221 ) induced in P29320 293 cells was also inhibited by P60568 . We also found that P60568 differentially decreased the peak amplitudes of Q12879 - and Q13224 -containing DB01221 receptor-mediated currents ( I( Q12879 ) and I( Q13224 ) ) by 54+/-5 % and 30+/-4 % , respectively . These results provide new evidence that P60568 induces rapid inhibition of peak currents of DB01221 receptor-mediated responses with possible Q9UHB4 / Q12879 and Q9UHB4 / Q13224 subtype-differentiation , and suggest that the inhibition is mediated by direct interaction between P60568 and DB01221 receptors . [ DB01407 in amyotrophic lateral sclerosis. No indication for a positive effect ] . The anabolic effects of clenbuterol have been recognized for a long time . DB01407 augments the expression of specific muscle proteins with a differential effect on type I and type II fibres . Furthermore , clenbuterol induces the synthesis of endogenous nerve growth factor ( P01138 ) and may itself be a myotrophic factor released by neuron endings . Side effects include tremor and headache and dose dependent abnormalities of laboratory values ( hypokalemia , hypoglycemia ) . After long-term medication increasing fatigue of muscles has been observed . Decreased expression of beta 2-adrenergic receptors may limit the expected functional improvement . The efficacy of clenbuterol as symptomatic treatment of amyotrophic lateral sclerosis has not been proved . Controlled treatment trials are warranted to assess this question . Treatment with arimoclomol , a coinducer of heat shock proteins , delays disease progression in P35858 mice . Amyotrophic lateral sclerosis ( P35858 ) is a fatal neurodegenerative condition in which motoneurons of the spinal cord and motor cortex die , resulting in progressive paralysis . This condition has no cure and results in eventual death , usually within 1-5 years of diagnosis . Although the specific etiology of P35858 is unknown , 20 % of familial cases of the disease carry mutations in the gene encoding Cu/Zn superoxide dismutase-1 ( P00441 ) . Transgenic mice overexpressing human mutant P00441 have a phenotype and pathology that are very similar to that seen in human P35858 patients . Here we show that treatment with arimoclomol , a coinducer of heat shock proteins ( HSPs ) , significantly delays disease progression in mice expressing a P00441 mutant in which glycine is substituted with alanine at position 93 ( P00441 (G93A) ) . DB05025 -treated P00441 (G93A) mice show marked improvement in hind limb muscle function and motoneuron survival in the later stages of the disease , resulting in a 22 % increase in lifespan . Pharmacological activation of the heat shock response may therefore be a successful therapeutic approach to treating P35858 , and possibly other neurodegenerative diseases . Chronic methamphetamine exposure alters immune function in normal and retrovirus-infected mice . Methamphetamine ( MA ) abuse represents a growing problem in the USA with an increase of sudden death . To evaluate the immune function alterations due to chronic methamphetamine use , we examined C57BL/C mice with LP-BM5 retrovirus infection plus methamphetamine exposure . Mice were randomly assigned to the following groups : placebo , placebo retrovirus-infected , uninfected MA treated and retrovirus-infected MA treated . Placebo , MA-treated groups were intraperitoneally injected with saline , MA , respectively , with a gradually increasing dose from 15 to 40 mg/kg for 12 weeks ( 5 days/week ) . Con A- and LPS-induced mitogenesis of splenocytes , cytokine production by splenocytes culture and lipid peroxides in the liver were measured . Heart tissue histopathology was analyzed in all the groups with murine cytomegalovirus ( CMV ) superinfection . Our data showed that MA treatment significantly decreased production of P60568 and interferon gamma ( P01579 ) in uninfected mice but did not further suppress the reduced Th1 cytokines in retrovirus-infected mice . There were no significant effects on cytokines P05112 and P05231 . However , tumor necrosis factor ( P01375 ) was significantly increased in both uninfected and infected mice due to MA treatment . Lipid peroxides in liver were significantly increased both in uninfected and retrovirus-infected mice due to MA exposure . DB00163 levels in liver were significantly decreased in uninfected mice due to MA treatment . CMV superinfection greatly increased the cardiac lesions in retrovirus-infected mice while no significant histopathology changes were detected due to MA treatment . Our data suggest that MA has immunomodulation activity , suppressing Th1 cytokine production and enhancing some Th2 cytokine secretion , as well as increasing lipid peroxides in uninfected mice . The interaction between LP-BM5 and MA remains unclear . Synthesis and biological evaluation of novel ( 4 or 5-aryl ) pyrazolyl-indoles as inhibitors of interleukin-2 inducible T-cell kinase ( Q08881 ) . P60568 inducible T-cell kinase ( Q08881 ) is one of five kinases that belong to the Tec kinase family that plays an important role in T-cell and mast cell signaling . Various reports point to a role of Q08881 in the treatment of allergic asthma . For example , it was shown that mice lacking Q08881 have reduced airway hyperresponsiveness , inflammation and tracheal responses in an allergic asthma model . In this article , we disclose novel Q08881 inhibitors based on ( 4 or 5-aryl ) pyrazolyl-indole scaffold that were also found to be selective for Q08881 over other kinases like IRK , P24941 , GSK3ss and PKA .

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interacts_with DB00227?

multiple_choice

Antifolate resistance due to new and known Plasmodium falciparum dihydrofolate reductase mutations expressed in yeast . Two new dihydrofolate reductase ( P00374 ) mutations were recently discovered in Plasmodium falciparum samples from an area of Bolivia with high rates of in vivo resistance to pyrimethamine-sulfadoxine : a DB00151 --> DB00125 point mutation in codon 50 and a five amino acid insertion after codon 30 , termed the Bolivia repeat . We used a yeast expression system to screen these new P00374 mutants , as well as all of the other known P00374 mutant genotypes , against four antifolates : pyrimethamine , cycloguanil , chlorcycloguanil , and WR99210 . The prodrug proguanil was also evaluated . The primary 108- DB00174 mutation , the known secondary mutations 51- DB00167 , 59- DB00125 and 164- DB00149 , as well as the 50- DB00125 mutation , all progressively enhanced pyrimethamine resistance in naturally observed combinations with one another , with the presence of 164- DB00149 most significantly increasing resistance . Cycloguanil and chlorcycloguanil resistance were most impacted by 164- DB00149 and the paired 16- DB00161 /108- DB00156 . DB01131 had no effect on malaria P00374 . All DHFRs analyzed were sensitive to WR99210 . The Bolivia repeat did not markedly affect drug sensitivity . We conclude that malaria P00374 can be reliably , rapidly and inexpensively analyzed in yeast for activity against a broad spectrum of antifolates . This system may be useful for initially characterizing newly discovered genotypes before proceeding to P. falciparum transfection ; for large-scale geographic surveys of drug resistance ; and for screening new antifolates or new antifolate combinations for their effectiveness against a large panel of P00374 mutants . DB09341 transporter-2 ( P11168 ) promoter mediated transgenic insulin production reduces hyperglycemia in diabetic mice . P01308 production afforded by hepatic gene therapy ( HGT ) retains promise as a potential treatment for type 1 diabetes , but successful approaches have been limited . We employed a novel and previously untested promoter for this purpose , glucose transporter-2 ( P11168 ) to drive insulin production via delivery by recombinant adeno-associated virus ( rAAV ) . In vitro , the P11168 promoter was capable of robust glucose-responsive expression in transduced HepG2 human hepatoma cells . Therefore , rAAV constructs were designed to express the furin-cleavable human preproinsulin B10 gene , under the control of the murine P11168 promoter and packaged for delivery with rAAV expressing the type 5 capsid . DB00428 -induced diabetic mice were subjected to hepatic portal vein injection immediately followed by implantation of a sustained-release insulin pellet to allow time for transgenic expression . All mice injected with the rAAV5- P11168 -fHPIB10 virus remained euglycemic for up to 35 days post-injection , with 50 % euglycemic after 77 days post-injection . In contrast , mock-injected mice became hyperglycemic within 15 days post-injection following dissolution of the insulin pellet . Serum levels of both human insulin and C-peptide further confirmed successful transgenic delivery by the rAAV5- P11168 -fHPIB10 virus . These findings indicate that the P11168 promoter may be a potential candidate for regulating transgenic insulin production for hepatic insulin gene therapy in the treatment of type I diabetes . Adipose tissue tumor necrosis factor and interleukin-6 expression in human obesity and insulin resistance . Adipose tissue expresses tumor necrosis factor ( P01375 ) and interleukin ( IL ) -6 , which may cause obesity-related insulin resistance . We measured P01375 and P05231 expression in the adipose tissue of 50 lean and obese subjects without diabetes . P01308 sensitivity ( S(I) ) was determined by an intravenous glucose tolerance test with minimal-model analysis . When lean [ body mass index ( BMI ) < 25 kg/m(2) ] and obese ( BMI 30-40 kg/m(2) ) subjects were compared , there was a 7.5-fold increase in P01375 secretion ( P < 0.05 ) from adipose tissue , and the P01375 secretion was inversely related to S(I) ( r = -0.42 , P < 0.02 ) . P05231 was abundantly expressed by adipose tissue . In contrast to P01375 , plasma ( rather than adipose ) P05231 demonstrated the strongest relationship with obesity and insulin resistance . Plasma P05231 was significantly higher in obese subjects and demonstrated a highly significant inverse relationship with S(I) ( r = -0.71 , P < 0.001 ) . To separate the effects of BMI from S(I) , subjects who were discordant for S(I) were matched for BMI , age , and gender . By use of this approach , subjects with low S(I) demonstrated a 3.0-fold increased level of P01375 secretion from adipose tissue and a 2.3-fold higher plasma P05231 level ( P < 0.05 ) compared with matched subjects with a high S(I) . Plasma P05231 was significantly associated with plasma nonesterified fatty acid levels ( r = 0.49 , P < 0.002 ) . Thus the local expression of P01375 and plasma P05231 are higher in subjects with obesity-related insulin resistance . Maturation of dendritic cells by recombinant human P29965 -trimer leads to a homogeneous cell population with enhanced surface marker expression and increased cytokine production . Dendritic cells ( DC ) have been shown to be potent inducers of specific cytotoxic T-cell responses both in vivo and in vitro . Furthermore , exposure to cytokines such as tumour necrosis factor ( P01375 ) -alpha or P25942 triggering changes DC phenotype and cytokine production and may enhance the T-cell activating capacity of the DC . We studied DC phenotype and cytokine production as well as the T-cell proliferation and cytotoxic T lympocyte ( CTL ) activation induced by DC generated in vitro . In addition , the effect of exposure to recombinant human P29965 -trimer ( huCD40LT ) on these parameters was investigated . Effective differentiation of monocytes derived from freshly isolated peripheral blood mononuclear cells ( PBMC ) was obtained with granulocyte macrophage-colony stimulating factor ( GM- P04141 ) and interleukin ( IL ) -4 . The DC expression of human leucocyte antigen ( HLA ) molecules , P33681 , Q01151 , and P42081 was markedly enhanced by exposure to huCD40LT even compared to P01375 exposure . Only a moderate cytokine production was observed initially , while P01375 addition or P25942 triggering , especially , induced enhanced production of P05231 and IL-12 p40 . Surprisingly , comparable induction of T-cell proliferation by a DC allostimulus or through the presentation of PPD , and influenza M1-peptide specific CTL activity was obtained with nonmaturated ( Q01151 - ) and maturated ( Q01151 + ) DC . In conclusion , a final maturation of monocyte-derived DC through huCD40LT resulted in a highly homogeneous cell population with enhanced surface marker expression and high production of pro-inflammatory cytokines . In addition , the induction of responses to allo or recall antigens presented by huCD40LT maturated DC was comparable to the responses obtained with the DC maturated through P01375 exposure . 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 . Endothelial dysfunction in congestive heart failure : P12821 inhibition vs. angiotensin II antagonism . BACKGROUND : Endothelial dysfunction of the vasculature contributes to the elevated peripheral resistance and reduced myocardial perfusion in congestive heart failure ( CHF ) . The present study systematically investigated the effect of angiotensin II ( AT(1) ) - receptor blockade on vascular superoxide ( O(2)(-) ) production and endothelial dysfunction . METHODS AND RESULTS : Vasodilator responses and O(2)(-) production were determined in aortic rings from Wistar rats with experimental CHF 10 weeks after extensive myocardial infarction and compared with sham-operated animals ( Sham ) . Rats were either treated with placebo ( P ) , with the AT(1)-receptor antagonist Irbesartan ( 50 mg kg(-1) day(-1) ) or with the P12821 inhibitor DB00519 ( 0.3 mg kg(-1) day(-1) ) . In CHF-P , endothelium-dependent , acetylcholine-induced relaxation was significantly attenuated compared with Sham-P . Chronic treatment with DB00519 or Irbesartan significantly improved endothelium-dependent relaxation . Aortic O(2)(-) formation was markedly increased in CHF , and was not significantly affected by DB00519 treatment , while it was reduced by Irbesartan. P29474 expression was reduced in CHF and normalised by both treatments . CONCLUSION : Endothelial vasomotor function in CHF rats was normalised by long-term treatment with an P12821 inhibitor or an AT(1)-antagonist . Reduced aortic P29474 expression was normalised by both treatments , whereas aortic superoxide formation was only reduced by the AT(1)-antagonist Irbesartan . 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 . 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 . DB04901 ( anti- P33681 ) -induced growth inhibition and prolongation of survival in vivo of B- Q9NZ71 tumor xenografts and potentiation by the combination with fludarabine . DB04901 is a primatized monoclonal antibody that targets P33681 expressed on malignant B cells and is being studied in the clinic as a potential treatment for follicular Q9NZ71 . We have recently reported that galiximab signals B- Q9NZ71 cells in vitro and inhibits cell growth and sensitizes resistant tumor cells to apoptosis by chemotherapeutic drugs . This study was designed to validate the in vitro findings in in vivo in mice . Thus , we examined in vivo the antitumor activity of galiximab used alone and in combination with chemotherapeutic agents in SCID mice bearing human lymphoma xenografts . The in vivo antitumor effects of galiximab used alone and in combination with fludarabine or doxorubicin were determined in solid and disseminated human B-lymphoma tumors grown in SCID mice . DB04901 monotherapy in vivo demonstrated significant antitumor activity in a Raji lymphoma solid tumor model and in an SKW disseminated lymphoma tumor model . There was significant inhibition in tumor growth and prolongation of survival . In vitro , galiximab sensitized Raji cells to apoptosis by both fludarabine and doxorubicin . Tumor growth inhibition was significantly enhanced when the mice were treated with the combination of galiximab and fludarabine . These findings support the potential clinical application of galiximab in combination with chemotherapeutic drugs for the treatment of P33681 -expressing hematological malignancies . 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 . 17 DB00783 overcomes a P55008 block induced by P04035 inhibitors and fosters cell cycle progression without inducing P27361 and -2 Q96HU1 kinases activation . P04035 inhibitors , such as DB00227 and Simvastatin , cause cell cycle arrest by interfering with the mitogenic activity of mitogens present in culture media . Cells are induced to pause in P55008 and can readily resume growth upon removal of the enzymatic block . DB00286 , acting via their nuclear receptor , are mitogens for different normal and transformed cell types , where they foster cell cycle progression and cell division . In estrogen-responsive MCF-7 human breast cancer cells , but not in non responsive cells , 17 beta-estradiol ( E2 ) induces cells arrested with DB00227 or Simvastatin to proliferate in the presence of inhibitor , without restoring P04035 activity or affecting the protein prenylation pattern . Mitogenic stimulation of P55008 -arrested MCF-7 cells with E2 includes primary transcriptional activation of c-fos , accompanied by transient binding in vivo of the estrogen receptor and/or other factors to the ERE and the estrogen-responsive DNA region of this proto-oncogene , as detected by dimethylsulphate genomic footprinting analysis . Mitogenic stimulation of growth-arrested MCF-7 cells by E2 occurs , under these conditions , without evident activation of P27361 and -2 kinases , and thus independently from the mitogen-responsive signal transduction pathways that converge on these enzymes . 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 . 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 . 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 . Inhibitory effect of novel 5-O-acyl juglones on mammalian DNA polymerase activity , cancer cell growth and inflammatory response . We previously found that vitamin K(3) ( menadione , 2-methyl-1,4-naphthoquinone ) inhibits the activity of human mitochondrial DNA polymerase γ ( pol γ ) . In this study , we focused on juglone ( 5-hydroxy-1,4-naphthoquinone ) , which is a 1,4-naphthoquinone derivative , and chemically synthesized novel juglones conjugated with P06681 :0 to C22:6 fatty acid ( 5-O-acyl juglones ) . The chemically modified juglones enhanced mammalian pol inhibition and their cytotoxic and anti-inflammatory activities . The juglone conjugated with oleic acid ( C18:1-acyl juglone ) showed the strongest inhibition of DNA replicative pol α activity and human colon carcinoma ( HCT116 ) cell growth in 10 synthesized 5-O-acyl juglones . C12:0-Acyl juglone was the strongest inhibitor of DNA repair-related pol λ , as well as the strongest suppression of the production of tumor necrosis factor ( P01375 ) -α production induced by lipopolysaccharide ( LPS ) in the compounds tested . Moreover , this compound caused the greatest reduction in 12-O-tetradecanoylphorbol-13-acetate ( TPA ) -induced acute inflammation in mouse ears . C12:0- and C18:1-Acyl juglones selectively inhibited the activities of mammalian pol species , but did not influence the activities of other pols and DNA metabolic enzymes tested . These data indicate that the novel 5-O-acyl juglones target anti-cancer and/or anti-inflammatory agents based on mammalian pol inhibition . Moreover , the results suggest that acylation of juglone is an effective chemical modification to improve the anti-cancer and anti-inflammation of vitamin K(3) derivatives , such as juglone . P01375 inhibits flow and insulin signaling leading to NO production in aortic endothelial cells . Endothelial cells release nitric oxide ( NO ) acutely in response to increased " flow " or fluid shear stress ( FSS ) , and the increase in NO production is correlated with enhanced phosphorylation and activation of endothelial nitric oxide synthase ( P29474 ) . Both vascular endothelial growth factor and FSS activate endothelial protein kinase B ( P31749 ) by way of incompletely understood pathway(s) , and , in turn , P31749 phosphorylates P29474 at DB00133 -1179 , causing its activation . In this study , we found that either FSS or insulin stimulated insulin receptor substrate-1 ( P35568 ) tyrosine and serine phosphorylation and increased P35568 -associated phosphatidylinositol 3-kinase activity , phosphorylation of P31749 DB00133 -473 , phosphorylation of P29474 DB00133 -1179 , and NO production . Brief pretreatment of bovine aortic endothelial cells with tumor necrosis factor-alpha ( P01375 ) inhibited the above described FSS- or insulin-stimulated protein phosphorylation events and almost totally inhibited FSS- or insulin-stimulated NO production . These data indicate that FSS and insulin regulate P29474 phosphorylation and NO production by overlapping mechanisms . This study suggests one potential mechanism for the development of endothelial dysfunction in disease states with alterations in insulin regulation and increased P01375 levels . 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 . In vitro and in vivo biological activities of a novel nonpolyglutamable anti-folate , MX-68 . MX-68 is a newly synthesized anti-folate , chemically designed not to undergo intracellular polyglutamation and to have increased affinity to dihydrofolate reductase ( P00374 ) . In the present study , we examined the in vitro and in vivo biological activities of MX-68 compared with methotrexate ( MTX ) which forms several polyglutamates intracellularly . MX-68 dose-dependently inhibited the proliferation of PHA- , anti-CD3- , or PMA plus ionomycin-stimulated peripheral blood mononuclear cells ( PBMC ) and endothelial cells ( EC ) from normal subjects as well as P01584 - or P01375 alpha-stimulated synovial fibroblastic cells ( SC ) from rheumatoid arthritis ( RA ) patients . Coaddition of folinic acid completely reversed the anti-proliferative effects of both MX-68 and MTX . Although the anti-proliferative activities of MX-68 were almost comparable to those of MTX , the washout study clearly showed the characteristic nature of MX-68 . When drugs were removed during culture , the suppressive effect of MX-68 completely disappeared , whereas suppression by MTX was merely weakened . MX-68 dramatically suppressed the onset of collagen-induced arthritis ( CIA ) in mice when the drug was orally administered three times a week. starting from the day of first immunization . In this model , 2 mg/kg of MX-68 was sufficient to completely suppress arthritis , whereas suppression by the same dose of MTX was partial . These lines of evidence suggest that polyglutamation is not always a prerequisite in the anti-rheumatic effects of anti-folate . In addition , since intracellular accumulation of polyglutamates is thought to have adverse effects , MX-68 may become a more potent and less toxic anti-rheumatic drug than MTX . Evaluation of antitumour activity of tea carbohydrate polymers in hepatocellular carcinoma animals . Box-Behnken design criterion was applied to identify the significant effects of various extraction parameters such as temperature , time , and solvent-solid ratio on extraction of tea carbohydrate . Among the three variables tested extraction temperature , and solvent-solid ratio were found to have significant effect on tea carbohydrate extraction . The most suitable condition for extraction of tea carbohydrate was found to be a single step extraction at extraction temperature 90°C , extraction time 30 min , and solvent-solid ratio 5:1 . At these optimum extraction parameters , the maximum yield of tea carbohydrate obtained experimentally was found to be very close to its predicted value of 3.47 % dry weight of root . Then , we have studied the influence of tea carbohydrate on biochemical parameters in hepatocellular carcinoma ( HCC ) animals . Hepatocellular carcinoma was induced by the injection of 1×10(5) H22 hepatocarcinoma cells into right hind thigh muscle in experimental animals . Tea carbohydrate could inhibit tumour growth and decrease microvessel density in tumour tissue . The altered amount of serum white blood cells ( WBC ) , Interferon-gamma ( IFN-γ ) and tumor necrosis factor-alpha ( P01375 -α ) in HCC animals were dose-dependently increased , whereas activities of serum alanine transaminase ( ALT ) , aspartate transaminase ( Q9NRA2 ) and alkaline phosphatase ( ALP ) were dose-dependently decreased in the drug treated animals . In addition , tea carbohydrate administration could decrease expression of vascular endothelial growth factor ( P15692 ) and proliferating cell nuclear antigen ( P12004 ) in H22 tumor tissue . It can be concluded that tea carbohydrate displayed strong antitumour activity in animals . [ DB00594 attenuates hypoxia-induced proliferation of rats pulmonary artery smooth muscle cells by suppressing Na+/ H+ exchanger-1 ] . AIM : To study the influence of Na+/H+ exchange inhibitor amiloride on hypoxia-induced proliferation in rats pulmonary artery smooth muscle cells ( PASMCs ) , also observe the change of Na+/H+ exchanger-1 ( P19634 ) activity and expression . METHODS : Rats PASMGs were cultured in normoxia ( 21 % O2 ) or hypoxia ( 2 % O2 ) for 24 hours , as well as administered amiloride with various concentrations , cultured for 24 hours , then determined MTT OD values and rates of P12004 positive cells to investigate cells proliferation , moreover intracellular pH was determined by interactive Laser Cytometer , and Na+/H+ exchanger-1 mRNA expression was determined by RT-PCR . RESULTS : Hypoxic exposure heightened intracellular pH and mRNA expression of P19634 in PASMCs , however , 3.123-50 micromol/L amiloride depressed them gradually . Additionally , hypoxic exposure raised MTT OD value and rates of P12004 positive cells , similarly , the above two indexes descended gradually with presence of 3.125-50 micromol/L amiloride . CONCLUSION : Na+/H+ exchange inhibitor amiloride can suppress hypoxia-induced proliferation in pulmonary artery smooth muscle cells , which is due to depress activity and expression of P19634 .

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interacts_with DB00559?

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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 . 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 . Endothelial progenitor cells in relation to endothelin-1 and endothelin receptor blockade : a randomized , controlled trial . AIMS : Endothelial progenitor cells ( EPC ) represent an endogenous repair mechanism involving rendothelialization and neoangiogenesis . Patients with both diabetes and vascular disease have low numbers of circulating EPC . The endothelium-derived peptide , endothelin-1 ( ET-1 ) , is increased in patients with type 2 diabetes and vascular complications and has been suggested to contribute to endothelial dysfunction . Therefore , we investigated the relation between EPC and plasma ET-1 and the effect of dual ET-1 receptor antagonist treatment . METHODS : In this double blind study patients with type 2 diabetes mellitus and microalbuminuria were randomized to treatment with the dual P25101 /ETB receptor antagonist DB00559 treatment ( 125mg bid ; n=17 ) or placebo ( n=19 ) for four weeks . Different EPC subpopulations were enumerated by flow cytometry using triple staining ( P28906 , CD133 , P35968 ) at baseline at the end of treatment . Viability was assessed by 7AAD and Annexin-V-staining . RESULTS : Baseline ET-1 levels correlated significantly with P02741 levels . Patients with ET-1 levels above the median value had higher levels of P28906 (+)CD133(+) and P28906 (+) P35968 (+) EPC . There was no difference in P28906 (+) and P28906 (+)CD133(+) P35968 (+) cells , markers of EPC apoptosis or circulating markers of endothelial damage between patients with ET-1 levels below or above the median . Four week treatment with DB00559 did not change EPC levels . CONCLUSION : Among patients with type 2 diabetes and vascular disease , high plasma levels of ET-1 are associated with higher number of EPC . The recruitment of EPC does not seem to be regulated via ET-1 receptor activation since treatment with a dual ET-1 receptor blocker did not affect circulating EPC numbers . Q08462 selectively couples to E prostanoid type 2 receptors , whereas adenylyl cyclase 3 is not receptor-regulated in airway smooth muscle . Adenylyl cyclases ( ACs ) are important regulators of airway smooth muscle function , because β-adrenergic receptor ( βAR ) agonists stimulate AC activity and DB02527 production . We have previously shown in a number of cell types that AC6 selectively couples to βAR and these proteins are coexpressed in lipid rafts . We overexpressed AC2 , O60266 , and AC6 in mouse bronchial smooth muscle cells ( mBSMCs ) and human embryonic kidney ( P29320 ) -293 cells by using recombinant adenoviruses and assessed their localization and regulation by various G protein-coupled receptors ( GPCRs ) . O60266 and AC6 were expressed primarily in caveolin-rich fractions , whereas AC2 expression was excluded from these domains . AC6 expression enhanced DB02527 production in response to isoproterenol but did not increase responses to butaprost , reflecting the colocalization of AC6 with β(2)AR but not E prostanoid type 2 receptor ( EP(2)R ) in lipid raft fractions . AC2 expression enhanced butaprost-stimulated DB02527 production but had no effect on the β(2)AR-mediated response . O60266 did not couple to any GPCR tested . DB02587 -induced arborization of mBSMCs was assessed as a functional readout of DB02527 signaling . Arborization was enhanced by overexpression of AC6 and O60266 , but AC2 had no effect . GPCR-stimulated arborization mirrored the selective coupling observed for DB02527 production . With the addition of the phosphodiesterase 4 ( DB05876 ) inhibitor rolipram AC2 accelerated forskolin-stimulated arborization . Thus , AC2 selectively couples to EP(2)R , but signals from this complex are limited by DB05876 activity . O60266 does not seem to couple to GPCR in either mBSMCs or P29320 -293 cells , so it probably exists in a distinct signaling domain in these cells . Neovastat ( DB05387 ) inhibits the airway inflammation and hyperresponsiveness in a murine model of asthma . Matrix metalloproteinase ( MMP ) -9 plays an important role in the pathogenesis of bronchial asthma . Neovastat , having significant antitumor and antimetastatic properties , is classified as a naturally occurring multifunctional antiangiogenic agent . We evaluated the therapeutic effect of Neovastat on airway inflammation in a mouse model of asthma . BALB/c mice were immunized subcutaneously with ovalbumin ( OVA ) on days 0 , 7 , 14 , and 21 and challenged with inhaled OVA on days 26 , 29 , and 31 . Neovastat was administrated by gavage ( 5 mg/kg body weight ) three times with 12 h intervals , beginning 30 min before OVA inhalation . On day 32 , mice were challenged with inhaled methacholine , and enhanced pause ( Penh ) was measured as an index of airway hyperresponsiveness . The severity of airway inflammation was determined by differential cell count of bronchoalveolar lavage ( BAL ) fluid . The P14780 concentration in BAL fluid samples was measured by ELISA , and P14780 activity was measured by zymography . The untreated asthma group showed an increased inflammatory cell count in BAL fluid and Penh value compared with the normal control group . Mice treated with Neovastat had significantly reduced Penh values and inflammatory cell counts in BAL fluid compared with untreated asthmatic mice . Furthermore , mice treated with Neovastat showed significantly reduced P14780 concentrations and activity in BAL fluid . These results demonstrate that Neovastat might have new therapeutic potential for airway asthmatic inflammation . P25101 antagonists prevent the development of pulmonary emphysema in rats . We hypothesised that endothelin ( ET ) -1 plays an important role in the pathogenesis of emphysema . We attempted to apply ET-1 receptor antagonists to demonstrate and further elucidate the molecular pathogenesis pathways through which ET-1 may cause emphysematous changes . Sprague-Dawley rats were divided into four groups : control , cigarette smoke extract ( CSE ) , CSE+BQ-123 ( a selective endothelin receptor type A ( ET(A) ) antagonist ) and CSE+ DB00559 ( a mixed ET(A)/ET(B) receptor antagonist ) . The CSE was injected intraperitoneally once a week for 3 weeks , and BQ-123 or DB00559 was administered daily for the same duration . The expression of ET(A) receptor , apoptosis index , caspase-3 activity , matrix metalloproteinase ( MMP ) -2 and P14780 activity , and tumour necrosis factor ( P01375 ) -alpha and interleukin ( IL ) -1beta concentrations were measured in the lung tissue . The ET-1 levels and antioxidant activity were measured in the serum . Both BQ-123 and DB00559 prevented the development of CSE-induced emphysema , blocked the expression of ET(A) receptor , inhibited pulmonary apoptosis , inactivated P08253 and P14780 activities in the lung tissues , reduced the concentrations of inflammatory cytokines P01375 and IL-1beta , and improved the biological antioxidant activity in the serum . Emphysema development is suppressed by ET-1 receptor antagonists . ET-1 may cause emphysematous changes through molecular pathogenesis pathways involving apoptosis , proteinase and antiproteinase imbalance , inflammation and oxidative stress . Desensitization and internalization of endothelin receptor A : impact of G protein-coupled receptor kinase 2 ( P25098 ) -mediated phosphorylation . Endothelin receptor A ( P25101 ) , a G protein-coupled receptor , mediates endothelin signaling , which is regulated by P25098 . Three DB00133 and seven DB00156 residues recently proven to be phosphoacceptor sites are located in the C-terminal extremity ( CTE ) of the receptor following its palmitoylation site . We created various phosphorylation-deficient P25101 mutants . The phospholipase C activity of mutant receptors in P29320 -293 cells was analyzed during continuous endothelin stimulation to investigate the impact of phosphorylation sites on P25101 desensitization . Total deletion of phosphoacceptor sites in the CTE affected proper receptor regulation . However , proximal and distal phosphoacceptor sites both turned out to be sufficient to induce WT-like desensitization . Overexpression of the Gαq coupling-deficient mutant P25098 -D110A suppressed P25101 -WT signaling but failed to decrease phospholipase C activity mediated by the phosphorylation-deficient mutant P25101 -6PD . In contrast , P25098 -WT acted on both receptors , whereas the kinase-inactive mutant P25098 -D110A/K220R failed to inhibit signaling of P25101 -WT and P25101 -6PD . This demonstrates that P25101 desensitization involves at least two autonomous P25098 -mediated components : 1 ) a phosphorylation-independent signal decrease mediated by blocking of Gαq and 2 ) a mechanism involving phosphorylation of DB00133 and DB00156 residues in the CTE of the receptor in a redundant fashion , able to incorporate either proximal or distal phosphoacceptor sites . High level transfection of P25098 variants influenced signaling of P25101 -WT and P25101 -6PD and hints at an additional phosphorylation-independent regulatory mechanism . Furthermore , internalization of mRuby-tagged receptors was observed with P25101 -WT and the phosphorylation-deficient mutant P25101 -14PD ( lacking 14 phosphoacceptor sites ) and turned out to be based on a phosphorylation-independent mechanism . 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 . Immoderate inhibition of estrogen by anastrozole enhances the severity of experimental polyarthritis . P11511 inhibitors have become the standard of care for the adjuvant treatment of postmenopausal , hormone-sensitive breast cancer . Meanwhile , more and more breast cancer patients who are treated with aromatase inhibitors as adjuvant therapies often experience arthralgias and musculoskeletal aching , in some cases , have necessitated discontinuation of treatment . We therefore use a rat model of human RA to test the hypothesis that anastrozole , an aromatase inhibitor , could enhance arthritis . The parameters used for analyzing the disease severity included paw volume , radiology , histopathological examination , markers for cytokine profile , immunophenotypic assays , and immune response to type II collagen . Administration of anastrozole significantly increased the severity of arthritis . DB01217 induced the increased levels of proinflammatory cytokines , P01579 , IL-12 , and the decreased levels of P05112 , P22301 secretion . We further found that anastrozole suppressed the differentiation of naive T cells to Treg cells , and it blocked the balance of IgG2a/IgG1 in peripheral blood . Meanwhile , estradiol concentration was the lowest in the anastrozole group . In a well-established model of postmenopausal RA , anastrozole potently promote the progression of arthritis and the associated development of osteoporosis . This potential problem should alert the oncologists and other health professionals . Genetic polymorphisms in the P00797 -Angiotensin system in high-altitude and low-altitude Native American populations . Cardiovascular disease ( CVD ) is reportedly less common in high-altitude native populations than in lowlanders . To some extent , this is due to cultural and demographic factors ; however , increased cardiovascular efficiency contributing to hypoxia adaptation may also be involved . Numerous genetic variants have been associated with cardiovascular health . If the decreased incidence of CVD in modern high-altitude populations reflects selective pressures having favoured the transmission of these alleles in their antecedents , it would be expected that these alleles would be more common in highlanders than in lowlanders . We tested this hypothesis by determining the allele frequencies of five polymorphic loci in genes encoding components of the renin-angiotensin system ( DB01367 ) that have alleles associated with hypertension and cardiovascular disease in a high-altitude native Andean population , Quechua from the Peruvian altiplano , and in a lowland Amerindian population , Maya from the Yucatan peninsula . The polymorphisms examined were 1 ) the insertion/deletion polymorphism in intron 16 of the angiotensin converting enzyme ( P12821 ) gene ; 2 ) the A/G2350 transition ( P12821 -8 ) in intron 17 of the P12821 gene ; 3 ) the A/C1166 transversion in the 3' untranslated region of the angiotensin II receptor ( type 1 ) gene ( P30556 ) ; 4 ) the G/AI9-83 transition in intron 8 of the renin gene ( REN ) ; and 5 ) the T/C704 ( Met235Thr ) transition mutation in angiotensinogen ( AGT ) . There was no evidence for an over-representation of the DB01367 alleles associated with cardiovascular fitness in the high-altitude Amerindian population when compared to the lowland Amerindian population . In utero activation of fetal memory T cells alters host regulatory gene expression and affects HIV susceptibility . In utero priming to malaria antigens renders cord blood mononuclear cells ( CBMC ) more susceptible to productive HIV infection in vitro in the absence of exogenous stimulation . This provides a unique model to better understand mechanisms affecting lymphocyte susceptibility to HIV infection in vivo . Effector memory CD3(+) P01730 (+) T cells ( T(EM) ) were the exclusive initial targets of HIV with rapid spread to central memory cells . HIV susceptibility correlated with increased expression of CD25 and HLA-DR on T(EM) . Virus entered all samples equally , however gag/pol RNA was only detected in HIV susceptible samples , suggesting regulation of proviral gene transcription . Targeted analysis of human genes in memory T cells showed greater expression of P01579 , O95644 , P10914 , P01100 , and P62937 and decreased expression P25490 and Q12800 in HIV susceptible samples . Thus fetal priming to exogenous antigens enhances specific proviral gene transcription pathways in effector memory cells that may increase risk of vertical transmission of HIV . The complete mitochondrial genome sequence and gene organization of the mud crab ( Scylla paramamosain ) with phylogenetic consideration . The complete mitochondrial genome is of great importance for better understanding the genome-level characteristics and phylogenetic relationships among related species . In the present study , we determined the complete mitochondrial genome DNA sequence of the mud crab ( Scylla paramamosain ) by 454 deep sequencing and Sanger sequencing approaches . The complete genome DNA was 15,824 bp in length and contained a typical set of 13 protein-coding genes , 22 transfer RNA ( tRNA ) genes , two ribosomal RNA ( rRNA ) genes and a putative control region ( CR ) . Of 37 genes , twenty-three were encoded by the heavy strand ( H-strand ) , while the other ones were encoded by light strand ( L-strand ) . The gene order in the mitochondrial genome was largely identical to those obtained in most arthropods , although the relative position of gene tRNA( DB00117 ) differed from other arthropods . Among 13 protein-coding genes , three ( ATPase subunit 6 ( P00846 ) , DB00157 dehydrogenase subunits 1 ( P03886 ) and P03897 ) started with a rare start codon ATT , whereas , one gene cytochrome c oxidase subunit I ( P00395 ) ended with the incomplete stop codon TA . All 22 tRNAs could fold into a typical clover-leaf secondary structure , with the gene sizes ranging from 63 to 73 bp . The phylogenetic analysis based on 12 concatenated protein-coding genes showed that the molecular genetic relationship of 19 species of 11 genera was identical to the traditional taxonomy . 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 . Respective role of humoral factors and blood pressure in aortic remodeling of DOCA hypertensive rats . Hypertension results in increased thickness and stiffness of large artery walls . The goal of our study was to assess the respective roles of humoral factors such as Ang II , endothelin and blood pressure in these aortic modifications . For this purpose , uninephrectomized rats received DOCA and high salt diet , and when hypertension was installed , they were treated for 5 weeks with either a long-acting calcium antagonist , mibefradil ( 30 mg/kg/day ) , an P12821 inhibitor , enalapril ( 3 mg/kg/day ) , or a mixed P25101 and ETB endothelin receptor antagonist , DB00559 ( 100 mg/kg/day ) . A group of hypertensive rats was left untreated and a sham-operated group of normotensive rats was used for control . At the end of treatment , aortic medial thickness and elastin as well as collagen were evaluated by quantitative morphometry . DOCA-salt hypertensive rats exhibited a marked increase in medial thickness associated with no change in absolute content in extracellular matrix . P15502 relative density decreased in DOCA rats . Enalapril had no effect on arterial pressure . DB00559 decreased slightly ( by 12 mm Hg ) , but not significantly , blood pressure . None of these drugs had an effect on medial thickness suggesting that in DOCA hypertensive rats neither Ang II nor endothelin play a significant role in the remodeling of the aorta . In contrast , mibefradil almost normalized arterial pressure , prevented medial hypertrophy and increased elastin density . Further studies are required in order to assess if this effect is directly linked to the blood pressure decrease or to another mechanism related to the calcium antagonistic property of mibefradil . P10275 and invasion in prostate cancer . Activation of androgen receptor ( AR ) stimulates the growth of not only androgen-dependent but also of androgen-refractory prostate cancer . However , neither the role of AR in invasion/metastasis nor the relationship between invasiveness and androgen-refractory status has been established . In this study , we used the androgen-dependent prostate cancer cell line MDA PCa 2b , derived from a human bone metastasis , to generate an invasive subline ( MDA-I ) using a Matrigel chamber . MDA-I cells expressed higher levels of AR and prostate-specific antigen than their less invasive parental cells . Blocking AR function or removal of androgen suppressed the invasion of MDA-I cells , whereas stimulating AR increased invasion . In addition , forced AR overexpression increased the invasiveness of MDA PCa 2b cells . Next , we showed that an androgen-refractory subline ( MDA-hr ) of MDA PCa 2b cells also expressed higher levels of AR and were more invasive than their parental androgen-dependent cells . Blocking AR function suppressed the invasiveness of MDA-hr cells . Gelatin zymography indicated that matrix metalloproteinase 2 ( P08253 ) and P14780 activities were regulated by AR signaling and closely correlated with the invasiveness of the androgen-dependent and androgen-refractory prostate cancer cells . These data suggest that AR promotes the invasiveness of both androgen-dependent and androgen-refractory prostate cancer and that a more invasive phenotype might develop through AR activation during cancer progression . These findings potentially support the use of adjuvant hormonal therapy and the future development of more potent androgen blockade therapy . Comparison of effects of olmesartan and telmisartan on blood pressure and metabolic parameters in Japanese early-stage type-2 diabetics with hypertension . P30556 blockers ( ARBs ) are regarded as first-line treatments for type-2 diabetes with hypertension . Despite the availability of various types of ARBs , there are no comparative studies of their effects on patients with diabetes . In this open-label prospective crossover study , we compared the effects of olmesartan ( 20 mg/day ) and telmisartan ( 40 mg/day ) . Twenty Japanese early-stage type-2 diabetes patients with hypertension treated with valsartan ( 80 mg/day ) for at least 8 weeks were recruited to this study . At study entry , valsartan was changed to olmesartan ( 20 mg/day ) or telmisartan ( 40 mg/day ) and administered for 8 weeks . The drugs were then switched and treatment was continued for another 8 weeks . We analyzed the blood pressure lowering effects of each drug by 24-h ambulatory blood pressure monitoring at 0 , 8 , and 16 weeks . Simultaneously , we measured metabolic parameters and inflammation markers . DB00275 lowered mean systolic and diastolic blood pressure more significantly than did telmisartan . While there were no differences between the groups in metabolic parameters , including HbA1c and adiponectin , the decreases in serum interleukin-6 and highly sensitive P02741 were more significant by olmesartan treatment . Our results indicate that olmesartan has more potent arterial blood pressure lowering and anti-inflammatory effects than telmisartan . Soluble forms of P16070 and CD54 ( P05362 ) cellular adhesion molecules are released by human prostatic cancer cell lines . We have identified that human prostatic cancer cell lines DU145 , PC3 , P03886 , ALVA31 and JCA1 released soluble P16070 molecules and DU145 , PC3 and P03886 released soluble CD54 . Both soluble and surface P16070 were not found in LNCaP , and both forms of CD54 were not expressed in LNCaP and JCA1 . CD54 was found to be highly expressed on cell surface in ALVA31 , but these cells did not release soluble CD54 . Expression of both cell surface and soluble forms were examined after treatment of cells with P01579 , TGF-beta 1 , or culturing in serum-free media . The concentration of soluble CD54 in supernatants changed to small extent after treatment with TGF-beta 1 and increased after treatment with P01579 or in serum-free media . Cell surface expression of CD54 however , changed only minimally after treatment . The levels of cell surface and soluble P16070 also changed minimally after treatment with P01579 and TGF-beta 1 but decreased in serum-free media and this was accompanied by marked elevation of soluble P16070 in supernatants . These data indicate that soluble P16070 might be released from cell surface by shedding whereas alternative splicing is the most likely mechanism of soluble CD54 release . 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 . Comparison of human P25101 and ETB receptor signalling via G-protein and β-arrestin pathways . AIMS : To determine the pharmacology of ET(A)- and ET(B)-mediated β-arrestin recruitment and compare this to established human pharmacology of these receptors to identify evidence for endothelin receptor biased signalling and pathway specific blockade by antagonists . MAIN METHODS : The ability of ET-1 , P20800 , P14138 , sarafotoxin 6b and sarafotoxin 6c to activate ET(A) and ET(B)-mediated β-arrestin recruitment was determined in CHO- P04264 cells . Affinities were obtained for ET(A) selective ( BQ123 , sitaxentan , ambrisentan ) , ET(B) selective ( BQ788 ) and mixed ( DB00559 ) antagonists using ET-1 and compared to affinities obtained in competition experiments in human heart and by Schild analysis in human saphenous vein . Agonist dependence of affinities was compared for BQ123 and BQ788 in the ET(A) and ET(B) β-arrestin assays respectively . KEY FINDINGS : For β-arrestin recruitment , order of potency was as expected for the ET(A) ( ET-1≥ P20800 >> P14138 ) and ET(B) ( ET-1= P20800 = P14138 ) receptors . However , at the ET(A) receptor sarafotoxin 6b and P14138 were partial agonists . Antagonism of ET peptides by selective and mixed antagonists appeared non-competitive . BQ123 , but not BQ788 , exhibited agonist-dependent affinities . DB00559 was significantly more effective an inhibitor of β-arrestin recruitment mediated by ET(A) compared to the ET(B) receptor . In the ET(A) vasoconstrictor assay , ET-1 , P20800 and S6b were equipotent , full agonists and antagonists tested behaved in a competitive manner , although affinities were lower than predicted from the competition binding experiments in left ventricle . SIGNIFICANCE : These data suggest that the pharmacology of ET(A) and ET(B) receptors linked to G-protein- and β-arrestin mediated responses was different and DB00559 appeared to show bias , preferentially blocking ET(A) mediated β-arrestin recruitment . Dual endothelin receptor antagonism prevents remodeling of resistance arteries in diabetes . Vascular remodeling , characterized by extracellular matrix deposition and increased media-to-lumen ( M/L ) ratio , contributes to the development of microvascular complications in diabetes . We have previously shown in type 2 diabetic Goto-Kakizaki ( GK ) rats that selective P25101 receptor blockade prevents medial thickening of mesenteric arteries via regulation of matrix metalloproteases ( MMP ) , whereas selective ETB receptor blockade augments this thickening . The goal of this study was to determine the effect of combined P25101 and ETB receptor blockade on resistance vessel remodeling . Vessel structure , MMP activity , and extracellular matrix proteins were assessed in control Wistar and diabetic GK rats treated with vehicle or DB00559 ( 100 mg/kg per day ) for 4 weeks ( n = 7-9 per group ) . DB00559 completely prevented the increase in M/L ratio and P08253 activity in diabetes but paradoxically increased M/L ratio and MMP activation in control animals . Collagenase ( P45452 ) activity and protein levels were significantly decreased in diabetes . Accordingly , collagen deposition was augmented in GK rats . Dual ET receptor antagonism improved enzyme activity and normalized P45452 levels in diabetic animals but blunted P45452 activity in control animals . In summary , current findings suggest that diabetes-mediated remodeling of resistance arteries is prevented by dual blockade of P25101 and ETB receptors and that the relative role of ET receptors in the regulation of vascular structure differs in the control and disease states . Serial changes in serum vitamin P04264 , triglyceride , cholesterol , osteocalcin and 25-hydroxyvitamin D3 in patients after hip replacement for fractured neck of femur or osteoarthritis . Serum vitamin P04264 concentrations were measured at presentation ( just before surgery ) and then at weekly intervals for 3 weeks in two groups of elderly patients requiring either hemiarthroplasty for fractured neck of femur ( FON , n = 13 ) or total hip replacement for osteoarthritis of the hip ( OA , n = 16 ) . In comparison with healthy elderly volunteers ( n = 25 ) , serum vitamin P04264 concentrations were significantly lower in both groups at presentation , and fell significantly within 24 h after surgery to concentrations approaching non-detectable , subsequently returning to pre-operative values within 3 weeks . Serum vitamin P04264 tended to be lower in the fracture group both before and after operation , although calculation of a vitamin P04264 -triglyceride ratio reduced the apparent difference as triglyceride concentrations were lower in the fracture group . P02818 concentrations were similar and fell significantly after operation in both groups , returning to pre-operative levels within 7 days . No differences in the two forms of osteocalcin ( carboxylated and undercarboxylated ) were observed either before or after operation in either group . DB00146 concentrations were not significantly different between the two groups at any time . DB01022 status may be lower than desirable in certain groups of the elderly population , and supplementation should be considered as prophylactic therapy .

MH_train_1574

MH_train_1574

MH_train_1574

interacts_with DB00864?

multiple_choice

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 . Calcineurin-inhibitor-free immunosuppression based on the JAK inhibitor CP-690,550 : a pilot study in de novo kidney allograft recipients . This randomized , pilot study compared the Janus kinase inhibitor CP-690,550 ( 15 mg P55957 [ CP15 ] and 30 mg P55957 [ CP30 ] , n = 20 each ) with tacrolimus ( n = 21 ) in de novo kidney allograft recipients . Patients received an P60568 receptor antagonist , concomitant mycophenolate mofetil ( DB00688 ) and corticosteroids . CP-690,550 doses were reduced after 6 months . Due to a high incidence of BK virus nephropathy ( BKN ) in CP30 , DB00688 was discontinued in this group . The 6-month biopsy-proven acute rejection rates were 1 of 20 , 4 of 20 and 1 of 21 for CP15 , CP30 and tacrolimus groups , respectively . BKN developed in 4 of 20 patients in CP30 group . The 6-month rates of cytomegalovirus disease were 2 of 20 , 4 of 20 and none of 21 for CP15 , CP30 and tacrolimus groups , respectively . Estimated glomerular filtration rate was > 70 mL/min at 6 and 12 months ( all groups ) . NK cells were reduced by </= 77 % in CP-690,550-treated patients . In the CP-690,550 arms , there were modest lipid elevations and a trend toward more frequent anemia and neutropenia during the first 6 months . These data suggest that coadministration of CP-690,550 30 mg P55957 with DB00688 is associated with overimmunosuppression . At 15 mg P55957 , the efficacy/safety profile was comparable to the tacrolimus control group , excepting a higher rate of viral infection . Further dose-ranging evaluation of CP-690,550 is warranted . Effects of d-fenfluramine , MK-212 , and ondansetron on saline drinking in two-choice tests in the rehydrating rat . The aim of the present studies was to investigate the effects of serotonergic compounds on preference for isotonic saline and aversion to hypertonic saline , respectively . Twenty-two-hour water-deprived rats were divided into two groups : The first was given a choice between 0.9 % saline and water in a 30-min test ; the second was given a choice between 1.8 % saline and water . Animals were tested following administration of d-fenfluramine , the P28335 receptor agonist 6-chloro-2-(1-piperazinyl)pyrazine ( MK-212 ) , and the 5- Q9H205 receptor antagonist ondansetron . d- DB00574 ( 0.3-3.0 mg/kg ) did not reduce 0.9 % saline preference ; instead , at 0.3 mg/kg there was a significant increase in saline drinking . In contrast , MK-212 ( 0.3-3.0 mg/kg ) abolished the preference for isotonic saline whereas ondansetron ( 10-100 micrograms/kg ) had no effect . d- DB00574 and MK-212 reduced hypertonic saline drinking , although at the highest dose for each drug water drinking was also reduced . These data add further to the evidence for an important serotonergic involvement in the control of saline drinking and preference in the rat . 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 . Novel roles of Akt and P42345 in suppressing TGF-beta/ P36897 -mediated P84022 activation . P01308 -like growth factor-I inhibits transforming growth factor-beta ( TGF-beta ) signaling by blocking activation of P84022 ( S3 ) , via a phosphatidylinositol 3-kinase ( PI3K ) /Akt-dependent pathway . Here we provide the first report that the kinase activity of Akt is necessary for its ability to suppress many TGF-beta responses , including S3 activation and induction of apoptosis . Wild-type and myristoylated Akts ( Akt(WT) and Akt(Myr) ) suppress TGF-beta-induced phospho-activation of S3 but not Q15796 ( S2 ) , whereas kinase-dead Akt1 ( Akt1K179M ) or dominant-negative PI3K enhances TGF-beta-induced phospho-activation of both S2 and S3 . Using siRNA , rapamycin ( Rap ) , and adenoviral expression for P62942 -resistant and constitutively active P36897 ( P36897 ) , we demonstrate that mammalian target of Rap ( P42345 ) mediates Akt1 suppression of phospho-activation of S3 . These and further data on Akt1-S3 binding do not support a recently proposed model that Akt blocks S3 activation through physical interaction and sequestration of S3 from TGF-beta receptors . We propose a novel model whereby Akt suppresses activation of S3 in an Akt kinase-dependent manner through P42345 , a likely route for loss of tumor suppression by TGF-beta in cancers . 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 . 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 . P49190 -mediated inhibitory effect of parathyroid hormone and Q96A98 on cell proliferation . The parathyroid hormone ( PTH ) has dual mitogenic and inhibitory effects on cell proliferation , depending on the cell type and experimental conditions . PTH can signal via two different receptors , both positively coupled to the adenylyl cyclase/cyclic AMP pathway which can mimic some of the proliferative effects of PTH . We evaluated the role of the type-2 PTH ( PTH2 ) receptor on cell proliferation in clonal human embryonic kidney HEK293 cells stably expressing the human P49190 . Using a cyclic AMP-responsive gene-reporter , we confirmed that the tuberoinfundibular peptide ( Q96A98 ) and various human ( h ) PTH fragments including DB05829 -(1-34) were potent agonists ( EC(50) in the range of 0.01-0.3 nM ) whereas the bovine ( b ) PTH peptides b( DB00135 (34))PTH-(7-34) and its tryptophan derivative b[D- DB00150 (12), DB00135 (34)]PTH-(7-34) behaved as antagonists ( IC(50)=117 and 249 nM , respectively ) . DB05829 -(1-34) produced a dose-dependent inhibition of cell proliferation ( EC(50)=8.5+/-0.4 nM ) after 3 days and this effect was fully reversed by the tryptophan derivative antagonist . The same effect was observed with Q96A98 which caused a 30 % maximal inhibition . These findings reveal that P49190 activation can inhibit cell proliferation and might explain the dual functionality of PTH on cell proliferation . Low and high levels of DB00163 exert opposite effects on P60568 possibly through the modulation of P37231 , P25963 , and apoptotic pathway in activated splenocytes . OBJECTIVE : We previously demonstrated that a high dose of alpha-tocopheryl succinate inhibits interleukin-2 ( P60568 ) mRNA and production in autoimmune-prone MRL/lpr mice . In the present study , we investigated the regulation of DB00163 ( alphaTOC ) on P60568 gene expression by examining the mRNA of P60568 , inhibitor kappaBalpha ( P25963 ) , and peroxisome proliferator-activated receptor-gamma ( PPARgamma ) . METHODS : Messenger RNA expression in active splenocytes of BALB/c mice was investigated with reverse transcriptase polymerase chain reaction . RESULTS : Levels of P60568 mRNA in phorbol 12-myristate 13-acetate/ionomycin activated splenocytes and cytokine in T-helper-1 cells were increased by 50 microM of alphaTOC but decreased by 1 mM of alphaTOC . In addition , the P25963 gene expression significantly increased by the high dose ( > or=500 microM ) of alphaTOC , suggesting an inhibition on nuclear factor-kappaB pathway for activation of P60568 expression . PPARgamma mRNA level in activated splenocytes was upregulated by 1 mM of alphaTOC . PPARgamma mRNA level in unstimulated splenocytes was upregulated by alphaTOC in a dose-dependent manner , suggesting that alphaTOC might enhance the PPARgamma signaling pathway . High-dose alphaTOC induced apoptosis of splenocytes and inhibited phytohemagglutinin-stimulated T-cell proliferation . Conversely , the proliferative response of splenocytes was enhanced by 5 microM of alphaTOC . Low-dose ( 50 microM ) alphaTOC increased P60568 expression , which may have been due to the absence of downregulation of PPARgamma and P25963 on the P60568 gene . CONCLUSION : The results indicated that low and high doses of alphaTOC exert opposite effects on P60568 , possibly through modulation of PPARgamma , P25963 , and apoptosis pathways . The present findings support our previous observation of opposite effects of low- and high-dose vitamin E on survival of MRL/lpr mice . Human epidermal Langerhans ' cells are targets for the immunosuppressive macrolide tacrolimus ( FK506 ) . BACKGROUND : The immunosuppressive macrolide tacrolimus ( FK506 ) has been shown to inhibit allergic contact dermatitis in animal models as well as in human beings . More recently , successful treatment of atopic dermatitis with an ointment containing tacrolimus has been reported . OBJECTIVES : We explored the effects of this compound on epidermal Langerhans ' cells ( LCs ) , which are known to play an important pathophysiologic role in inflammatory skin diseases . METHODS : The expression of the intracellular FK506 binding protein ( P62942 ) was monitored on freshly isolated and cultured epidermal LCs . Phenotyping and functional exploration of LCs treated with different concentrations of tacrolimus and beta-methasone valerate ( betaMv ) were performed . RESULTS : P62942 is expressed in freshly isolated LCs but is lost while they are maturating into mature dendritic cells . DB00864 inhibited the expression of IL-2R ( CD25 ) and of the costimulatory molecules P33681 ( P33681 .1 ) and P25942 . Expression of MHC class I and II was also affected , whereas P42081 ( P33681 .2 ) expression was not altered . In contrast , betaMv strongly increased the expression of CD25 . Paradoxically , while decreasing P25942 and MHC class I expression , betaMv significantly increased the expression of MHC class II , P33681 , and P42081 on cultured LCs but impaired their allostimulatory activity . DB00864 was about 100 times more potent than betaMv at inhibiting LC stimulatory function . CONCLUSION : DB00864 can exert immunopharmacologic alterations on LCs , which may account , at least in part , for the therapeutic effect of this compound in eczematous skin diseases . Defective postnatal endochondral bone development by chondrocyte-specific targeted expression of parathyroid hormone type 2 receptor . The human parathyroid hormone type 2 receptor ( P49190 ) is activated by PTH and by tuberoinfundibular peptide of 39 residues ( Q96A98 ) , the latter likely acting as its natural ligand . Although the receptor is expressed at highest levels in the nervous system , we have observed that both P49190 and Q96A98 are expressed in the newborn mouse growth plate , with the receptor localizing in the resting zone and the ligand Q96A98 localizing exclusively in prehypertrophic and hypertrophic chondrocytes . To address the role of P49190 in postnatal skeletal growth and development , Col2a1-hPTH2R ( P49190 -Tg ) transgenic mice were generated . The mice were viable and of nearly normal size at birth . Expression of the transgene in the growth plate was limited to chondrocytes . We found that chondrocyte proliferation was decreased , as determined by in vivo BrdU labeling of proliferating chondrocytes and P11802 and P38936 expression in the growth plate of Col2a1-hPTH2R transgenic mice . Similarly , the differentiation and maturation of chondrocytes was delayed , as characterized by decreased Sox9 expression and weaker immunostaining for the chondrocyte differentiation markers collagen type II and type X and proteoglycans . As well , there was altered expression of Gdf5 , Wdr5 , and β-catenin , factors implicated in chondrocyte maturation , proliferation , and differentiation.These effects impacted on the process of endochondral ossification , resulting in delayed formation of the secondary ossification center , and diminished trabecular bone volume . The findings substantiate a role for P49190 signaling in postnatal growth plate development and subsequent bone mass acquisition . The specific Q14318 inhibitor N-(N',N'-dimethylcarboxamidomethyl)cycloheximide has potent neuroprotective and neurotrophic properties in brain ischemia . FK506 and FK506-derived inhibitors of the FK506-binding protein ( FKBP ) -type peptidylprolyl cis/trans-isomerases ( PPIase ) display potent neuroprotective and neuroregenerative properties in various neurodegeneration models , showing the importance of neuroimmunophilins as targets for the treatment of acute and chronic neurodegenerative diseases . However , the PPIase activity targeted by active site-directed ligands remains unknown so far . Here we show that neurotrophic FKBP ligands , such as GPI1046 and N-[methyl(ethoxycarbonyl)]cycloheximide , inhibit the calmodulin/Ca(2+) ( P62158 /Ca(2+) ) -regulated Q14318 with up to 80-fold higher affinity than P62942 . In contrast , the non-neurotrophic rapamycin inhibits Q14318 . P62158 /Ca(2+) 500-fold less affine than other neuroimmunophillins . In the context of the high expression of Q14318 in neuroblastoma cells , these data suggest that Q14318 . P62158 /Ca(2+) inhibition can mediate neurotrophic properties of FKBP ligands . The Q14318 -specific cycloheximide derivative , N-(N',N'-dimethylcarboxamidomethyl)cycloheximide ( DM-CHX ) was synthesized and used in a rat model of transient focal cerebral ischemia . Accordingly , DM-CHX caused neuronal protection as well as neural stem cell proliferation and neuronal differentiation at a dosage of 27.2 mug/kg . These effects were still dominant , if DM-CHX was applied 2-6 h post-insult . In parallel , sustained motor behavior deficits of diseased animals were improved by drug administration , revealing a potential therapeutic relevance . Thus , our results demonstrate that Q14318 inhibition by DM-CHX regulates neuronal cell death and proliferation , providing a promising strategy for the treatment of acute and/or chronic neurodegenerative diseases . 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 . 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 . 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 . P62942 , the 12-kDa FK506-binding protein , is a physiologic regulator of the cell cycle . P62942 , the 12-kDa FK506-binding protein , is a ubiquitous abundant protein that acts as a receptor for the immunosuppressant drug FK506 , binds tightly to intracellular calcium release channels and to the transforming growth factor beta ( TGF-beta ) type I receptor . We now demonstrate that cells from P62942 -deficient ( P62942 (-/-) ) mice manifest cell cycle arrest in G(1) phase and that these cells can be rescued by P62942 transfection . This arrest is mediated by marked augmentation of P38936 ( P38936 /CIP1) levels , which can not be further augmented by TGF-beta1 . The P38936 up-regulation and cell cycle arrest derive from the overactivity of TGF-beta receptor signaling , which is normally inhibited by P62942 . Cell cycle arrest is prevented by transfection with a dominant-negative TGF-beta receptor construct . TGF-beta receptor signaling to gene expression can be mediated by SMAD , p38 , and P29323 / Q96HU1 kinase ( extracellular signal-regulated kinase/mitogen-activated protein kinase ) pathways . SMAD signaling is down-regulated in P62942 (-/-) cells . Inhibition of P29323 / Q96HU1 kinase fails to affect P38936 up-regulation . By contrast , activated phosphorylated p38 is markedly augmented in P62942 (-/-) cells and the P38936 up-regulation is prevented by an inhibitor of p38 . Thus , P62942 is a physiologic regulator of cell cycle acting by normally down-regulating TGF-beta receptor signaling . Serotonin 2C receptor antagonists induce fast-onset antidepressant effects . Current antidepressants must be administered for several weeks to produce therapeutic effects . We show that selective serotonin 2C ( P28335 ) antagonists exert antidepressant actions with a faster-onset ( 5 days ) than that of current antidepressants ( 14 days ) in mice . Subchronic ( 5 days ) treatment with P28335 antagonists induced antidepressant behavioral effects in the chronic forced swim test ( cFST ) , chronic mild stress ( CMS ) paradigm and olfactory bulbectomy paradigm . This treatment regimen also induced classical markers of antidepressant action : activation of DB02527 response element-binding protein ( CREB ) and induction of brain-derived neurotrophic factor ( P23560 ) in the medial prefrontal cortex ( mPFC ) . None of these effects were induced by subchronic treatment with citalopram , a prototypical selective serotonin reuptake inhibitor ( SSRI ) . Local infusion of P28335 antagonists into the ventral tegmental area was sufficient to induce P23560 in the mPFC , and dopamine D1 receptor antagonist treatment blocked the antidepressant behavioral effects of P28335 antagonists . P28335 antagonists also activated mammalian target of rapamycin ( P42345 ) and eukaryotic elongation factor 2 ( eEF2 ) in the mPFC , effects recently linked to rapid antidepressant action . Furthermore , P28335 antagonists reversed CMS-induced atrophy of mPFC pyramidal neurons . Subchronic SSRI treatment , which does not induce antidepressant behavioral effects , also activated P42345 and eEF2 and reversed CMS-induced neuronal atrophy , indicating that these effects are not sufficient for antidepressant onset . Our findings reveal that P28335 antagonists are putative fast-onset antidepressants , which act through enhancement of mesocortical dopaminergic signaling . P62158 -mediated effects of loperamide on chloride transport by brush border membrane vesicles from human ileum . We investigated whether the synthetic opiate loperamide-HCl is able to regulate specific transport systems for sodium and chloride in brush border membrane vesicles ( BBMVs ) from human ileum and whether such activities are mediated by calcium/calmodulin . In BBMVs we studied Na+/H+ antiport , Cl+/OH- antiport , Na+/Cl- cotransport , and the Cl- conductive pathway . Brush border membrane vesicles were incubated with 10 microM loperamide over 4 h at 5 degrees C before the uptake experiments . In ileal BBMVs , loperamide stimulated intravesicular accumulation of Na+ in the presence of Cl- and vice versa . After 1 min of incubation , the stimulatory effect was 35 % +/- 5 % ( p less than 0.005 ) of the control without loperamide . DB00836 also stimulated Cl-/OH- antiport by 30 % +/- 5 % ( p less than 0.005 ) in BBMVs of ileum . In addition , we studied the role of Ca2+/calmodulin in the action of loperamide on chloride transport by human BBMVs . In loperamide-pretreated BBMVs , calmodulin activity was significantly decreased ( 12 +/- 2 vs. 38 +/- 4 pmol/mg protein ) . When loperamide-pretreated vesicles were incubated with 2 microM calcium ( free concentration ) plus 5 microM calmodulin for 1 h at 5 degrees C , complete inhibition of the stimulatory effect of loperamide on Cl-/OH- antiport and Na+/Cl- cotransport was observed . Increasing the Ca2+/calmodulin activity of loperamide-pretreated BBMVs with 2 microM calcium plus 5 microM calmodulin led to a significant inhibition of Cl-/OH- antiport and Na+/Cl- cotransport by 40 % +/- 5 % ( p less than 0.005 ) . 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 . Generation and phenotypic characterization of new human ovarian cancer cell lines with the identification of antigens potentially recognizable by HLA-restricted cytotoxic T cells . This study describes a simple method for long-term establishment of human ovarian tumor lines and prediction of T-cell epitopes that could be potentially useful in the generation of tumor-specific cytotoxic T lymphocytes ( CTLs ) . Nine ovarian tumor lines ( INT.Ov ) were generated from solid primary or metastatic tumors as well as from ascitic fluid . Notably all lines expressed HLA class I , intercellular adhesion molecule-1 ( P05362 ) , polymorphic epithelial mucin ( P15941 ) and cytokeratin ( CK ) , but not HLA class II , P33681 .1 ( P33681 ) or Q13072 . While of the 9 lines tested 4 ( INT.Ov1 , 2 , 5 and 6 ) expressed the folate receptor ( P15328 ) and 6 ( INT.Ov1 , 2 , 5 , 6 , 7 and 9 ) expressed the epidermal growth factor receptor ( P00533 ) ; MAGE-1 and p185HER-2/neu were only found in 2 lines ( INT.Ov1 and 2 ) and Q13065 expression in 1 line ( INT.Ov2 ) . The identification of class I MHC ligands and T-cell epitopes within protein antigens was achieved by applying several theoretical methods including : 1 ) similarity or homology searches to MHCPEP ; 2 ) BIMAS and 3 ) artificial neural network-based predictions of proteins MAGE , GAGE , P00533 , p185HER-2/neu and P15328 expressed in INT.Ov lines . Because of the high frequency of expression of some of these proteins in ovarian cancer and the ability to determine HLA binding peptides efficiently , it is expected that after appropriate screening , a large cohort of ovarian cancer patients may become candidates to receive peptide-based vaccines . 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 . Potential utility of telmisartan , an angiotensin II type 1 receptor blocker with peroxisome proliferator-activated receptor-gamma ( P37231 ) -modulating activity for the treatment of cardiometabolic disorders . The metabolic syndrome is strongly associated with insulin resistance and consists of a constellation of factors such as hypertension and hyperlipidemia that raise the risk for cardiovascular diseases and diabetes mellitus . There is widespread agreement that the renin-angiotensin system ( DB01367 ) plays a pivotal role in the pathogenesis of insulin resistance and cardiovascular disease in diabetes . Indeed , large clinical trials have demonstrated substantial benefit of the blockade of this system for cardiovascular end-organ protection . Thus the blockade of the DB01367 may be a promising strategy for the treatment of the patients with the metabolic syndrome . Although several types of angiotensin II type 1 ( AT(1) ) receptor blockers ( ARBs ) are commercially available for the treatment of patients with hypertension , we have recently found that telmisartan ( DB00966 ) could have the strongest binding affinity to AT(1) receptor . Further , telmisartan is reported to act as a partial agonist of peroxisome proliferator-activated receptor-gamma ( P37231 ) . These observations suggest that , due to its unique P37231 -modulating activity , telmisartan may be one of the most promising sartans for the treatment of cardiometabolic disorders . In this paper , we reviewed the potential utility of telmisartan in insulin resistance and vascular complications in diabetes . DB01599 inhibits O95477 -mediated cellular lipid efflux . OBJECTIVE : DB00171 -binding cassette transporter A1 ( O95477 ) mediates the efflux of lipids from cells to lipid-poor apolipoproteins . In this article , we characterize the effect of probucol on cellular O95477 -mediated lipid efflux . METHODS AND RESULTS : DB01599 inhibited cholesterol efflux up to 80 % in J774 macrophages expressing O95477 . In Fu5AH hepatoma cells that contain scavenger receptor class B , type I , but not functional O95477 , we observed no effect of probucol on cholesterol efflux . DB01599 inhibited cholesterol efflux from normal human skin fibroblasts but not from fibroblasts from a Tangier patient . Fluorescent confocal microscopy and biotinylation assay demonstrated that in J774 cells probucol impaired the translocation of O95477 from intracellular compartments to the plasma membrane . DB01599 also inhibited the formation of an O95477 -linked cholesterol oxidase sensitive plasma membrane domain . Consistent with the inhibitory effect on O95477 translocation to the plasma membrane , probucol reduced cell surface-specific [ 125I ] -labeled apolipoprotein-AI binding . CONCLUSIONS : We conclude that probucol is an effective inhibitor of O95477 -mediated cholesterol efflux without influencing scavenger receptor class B type I-mediated efflux . The inhibition of O95477 translocation to the plasma membrane may in part explain the reported in vivo high-density lipoprotein-lowering action of probucol .

MH_train_1575

MH_train_1575

MH_train_1575

interacts_with DB00623?

multiple_choice

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 . Modulation of β-catenin signaling by glucagon receptor activation . The glucagon receptor ( P47871 ) is a member of the class B G protein-coupled receptor family . Activation of P47871 by glucagon leads to increased glucose production by the liver . Thus , glucagon is a key component of glucose homeostasis by counteracting the effect of insulin . In this report , we found that in addition to activation of the classic DB02527 /protein kinase A ( PKA ) pathway , activation of P47871 also induced β-catenin stabilization and activated β-catenin-mediated transcription . Activation of β-catenin signaling was PKA-dependent , consistent with previous reports on the parathyroid hormone receptor type 1 ( Q03431 ) and glucagon-like peptide 1 ( P43220 ) receptors . Since low-density-lipoprotein receptor-related protein 5 ( Lrp5 ) is an essential co-receptor required for Wnt protein mediated β-catenin signaling , we examined the role of Lrp5 in glucagon-induced β-catenin signaling . Cotransfection with Lrp5 enhanced the glucagon-induced β-catenin stabilization and TCF promoter-mediated transcription . Inhibiting Lrp5/6 function using O94907 ( O94907 ) or by expression of the Lrp5 extracellular domain blocked glucagon-induced β-catenin signaling . Furthermore , we showed that Lrp5 physically interacted with P47871 by immunoprecipitation and bioluminescence resonance energy transfer assays . Together , these results reveal an unexpected crosstalk between glucagon and β-catenin signaling , and may help to explain the metabolic phenotypes of Lrp5/6 mutations . 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 . Anti- P04275 aptamer DB05202 for refractory thrombotic thrombocytopenic purpura . BACKGROUND : Plasma exchange is the main therapy for thrombotic thrombocytopenic purpura ( TTP ) . No treatments other than plasma exchange have been documented to be effective nor are approved for treatment of TTP . The anti- P04275 ( P04275 ) aptamer DB05202 effectively inhibits P04275 activity in plasma samples of TTP patients and thus shear-dependent platelet ( Q02083 ) function as measured by the Q02083 function analyzer PFA-100 ( Dade Behring ) . It was hypothesized that DB05202 would offer a potentially effective treatment option for a critically ill patient , refractory to standard care . CASE REPORT : A 39-year-old male patient with idiopathic TTP , refractory to daily plasma exchange , rituximab , steroids , and splenectomy , was additionally treated with a continuous infusion of the anti- P04275 ( P04275 ) aptamer DB05202 for 3 weeks . RESULTS : Plasma concentrations of approximately 10 microg/mL DB05202 decreased P04275 activity by more than 96 % . Plasma exchange treatment acutely decreased the plasma concentrations of DB05202 by a mean of 47 % ( range , 40 % -61 % ) . Thus , additional minibolus infusions of DB05202 were given after each plasma exchange to rapidly restore steady-state concentrations . DB05202 resulted in an increase of Q02083 counts as long as DB05202 was given . On three occasions the infusion was stopped , each accompanied by a decrease in Q02083 counts and worsening of microangiopathy . No serious adverse effects were observed during the treatment with DB05202 . CONCLUSION : DB05202 caused a clear and reproducible increase in Q02083 counts in an otherwise refractory TTP case . These clinical , pharmacokinetic , and pharmacodynamic data provide a rational basis for clinical trials with DB05202 in TTP . Temperature-dependent growth restriction in measles vaccine strains . Temperature-dependent growth restriction was studied with the measles vaccine strains licensed in Japan in comparison with their parental wild strains . Plaquing efficiency was compared at various temperatures from 35 to 40 C . O14965 -C strain derived from Edmonston wild strain was temperature-sensitive with the 39 C shutoff temperature . No significant restriction of growth was found for other vaccine strains , i.e. , Schwarz , FF-8 , and P62158 -70 , and for their parental wild strains , i.e. , Edmonston and Tanabe . A paradoxical feature was found for FF-8 strain ; in spite of undiminished plaquing efficiency at 40 C , the growth in the fluid medium was markedly depressed at 39 C or above . Development of treatment strategies for advanced neuroblastoma . Neuroblastoma is the most common cancer in childhood . The majority of patients with neuroblastoma are assigned to the high-risk group based on age at diagnosis , stage , histology , P04198 status , and DNA ploidy . Their prognosis remains unsatisfactory ; the 5-year event-free survival ( O43281 ) rate is generally 40 % . During the past 20 years , much effort has been made to reinforce chemotherapy , including the introduction of high-dose chemotherapy with autologous stem cell rescue , resulting in a 5-year O43281 rate of around 30 % . Subsequently , maintenance therapy aimed at eradicating residual tumors after induction and consolidation therapies was introduced , consisting of differentiation-inducing agents , retinoids , and immunotherapy using anti-GD2 antibodies combined with cytokines . However , such additional treatment provided benefit to only 10-20 % of patients , while the prognosis of about half the patients remains poor . Currently , novel targeted agents are under development . Among them , anaplastic lymphoma kinase ( Q9UM73 ) inhibitors and aurora kinase A inhibitors are promising . Q9UM73 somatic mutation or gene amplification predisposing neuroblastoma development occurs in up to 15 % of neuroblastomas . DB08865 is a dual-specific inhibitor of Q9UM73 / DB00134 and inhibits proliferation of neuroblastoma cells harboring R1275Q-mutated Q9UM73 or amplified wild-type Q9UM73 , but not cells harboring F1174L . Instead , cells with F1174L are sensitive to another small molecule Q9UM73 inhibitor , TAE684 . O14965 plays a pivotal role in centrosome maturation and spindle formation during mitosis . DB05220 ( alisertib ) is a small molecule inhibitor of aurora kinase A that is currently in early-phase clinical testing . Future treatment will be individually planned , adapting targeted agents based on personal biological tumor characteristics . 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 . DB05692 , a thrombin receptor ( P25116 ) antagonist for the prevention and treatment of atherothrombosis . DB05692 is a novel antiplatelet agent undergoing development by Schering-Plough Corp for the treatment and prevention of atherothrombosis . The compound is an orally administered himbacine analog that potently antagonizes the platelet thrombin receptor protease-activated receptor 1 ( P25116 ) , which leaves the procoagulant function of thrombin intact . In preclinical studies , DB05692 demonstrated no effect on bleed time or coagulation parameters . In both cynomolgus monkeys and humans , the compound had high bioavailability and inhibited ex vivo TRAP ( thrombin receptor-activating peptide ) -stimulated platelet aggregation in a potent and long-lasting manner . In a phase II clinical trial of patients undergoing percutaneous coronary intervention , DB05692 added to standard therapy with aspirin and clopidogrel did not increase major or minor thrombolysis in myocardial infarction bleeding , and demonstrated a trend toward decreased major adverse cardiovascular events versus placebo . At the time of publication , three phase III trials were underway to assess the efficacy and safety of DB05692 for at least 1 year in up to 35,000 patients with acute coronary syndromes or atherosclerosis . The distinct mechanism of action of DB05692 allows for cardiovascular protection without the liability of increased bleeding associated with other antiplatelet therapies . Phase III trials in high-risk patients will determine the use of DB05692 in cardiological practice . P04271 -mediated inhibition of the phosphorylation of P14136 is prevented by TRTK-12 . P04271 belongs to a family of calcium-binding proteins involved in cell cycle and cytoskeleton regulation . We observed an inhibitory effect of P04271 on glial fibrillary acidic protein ( P14136 ) phosphorylation , when stimulated by DB02527 or Ca2+/calmodulin , in a cytoskeletal fraction from primary astrocyte cultures . We found that P04271 has no direct effect on P62158 KII activity , the major kinase in this cytoskeletal fraction able to phosphorylate P14136 . The inhibition of P14136 phosphorylation is most likely due to the binding of P04271 to the phosphorylation sites on this protein and blocking the access of these sites to the protein kinases . This inhibition was dependent on Ca2+ . However , DB01593 could substitute for Ca2+ . The inhibitory effect of P04271 was prevented by TRTK-12 , a peptide that blocks P04271 interaction with several target proteins including glial fibrillary acidic protein . These data suggest a role for P04271 in the assembly of intermediate filaments in astrocytes . Store-operated Ca2+ entry ( SOCE ) induced by protease-activated receptor-1 mediates Q13586 protein phosphorylation to inhibit SOCE in endothelial cells through AMP-activated protein kinase and p38β mitogen-activated protein kinase . The Ca(2+) sensor Q13586 is crucial for activation of store-operated Ca(2+) entry ( SOCE ) through transient receptor potential canonical and Orai channels . Q13586 phosphorylation serves as an " off switch " for SOCE . However , the signaling pathway for Q13586 phosphorylation is unknown . Here , we show that SOCE activates AMP-activated protein kinase ( AMPK ) ; its effector p38β mitogen-activated protein kinase ( p38β MAPK ) phosphorylates Q13586 , thus inhibiting SOCE in human lung microvascular endothelial cells . Activation of AMPK using 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside ( AICAR ) resulted in Q13586 phosphorylation on serine residues and prevented protease-activated receptor-1 ( P25116 ) -induced Ca(2+) entry . Furthermore , AICAR pretreatment blocked P25116 -induced increase in the permeability of mouse lung microvessels . Activation of SOCE with thrombin caused phosphorylation of isoform α1 but not α2 of the AMPK catalytic subunit . Moreover , knockdown of AMPKα1 augmented SOCE induced by thrombin . Interestingly , SB203580 , a selective inhibitor of p38 MAPK , blocked Q13586 phosphorylation and led to sustained Q13586 -puncta formation and Ca(2+) entry . Of the three p38 MAPK isoforms expressed in endothelial cells , p38β knockdown prevented P25116 -mediated Q13586 phosphorylation and potentiated SOCE . In addition , inhibition of the SOCE downstream target P62158 kinase kinase β ( CaMKKβ ) or knockdown of AMPKα1 suppressed P25116 -mediated phosphorylation of p38β and hence Q13586 . Thus , our findings demonstrate that SOCE activates CaMKKβ-AMPKα1-p38β MAPK signaling to phosphorylate Q13586 , thereby suppressing endothelial SOCE and permeability responses . P62158 interacts with the cytoplasmic tails of the parathyroid hormone 1 receptor and a sub-set of class b G-protein coupled receptors . Parathyroid hormone ( PTH ) binds to its receptor ( PTH 1 receptor , Q03431 ) and activates multiple pathways . The Q03431 , a class b GPCR , contains consensus calmodulin-binding motifs . The Q03431 cytoplasmic tail interacts with calmodulin in a calcium-dependent manner via the basic 1-5-8-14 motif . DB01373 -dependent calmodulin interactions with the cytoplasmic tails of receptors for Q9Y3E5 , vasoactive intestinal peptide , pituitary adenylate cyclase activating peptide , corticotropin releasing hormone , calcitonin , and the glucagon-like peptides 1 and 2 are demonstrated . The cytoplasmic tails of the secretin receptor and the growth hormone releasing hormone receptor either interact poorly or not at all with calmodulin , respectively . DB00623 , a calmodulin antagonist , enhances PTH-mediated accumulation of total inositol phosphates , suggesting that calmodulin regulates signaling via phospholipase C . Arundic acid ( DB05343 ) ameliorates delayed ischemic brain damage by preventing astrocytic overproduction of P04271 . After focal cerebral ischemia , the infarct volume increases rapidly within acute infarct expansion ( initial 12 to 24 h ) and continues slowly during delayed infarct expansion ( 25 to 168 h ) . While acute infarct expansion represents progressive necrosis within the ischemic core , delayed infarct expansion starts as disseminated apoptotic cell death in a narrow rim surrounding the infarct border , which gradually coalesces to form a larger infarct . Discovery of a distinct correlation between reactive astrogliosis along the infarct border and delayed infarct expansion in the rodent ischemia model led us to investigate the possible causal relationship between the two events . Specifically , the calcium binding protein P04271 exerts detrimental effects on cell survival through activation of various intracellular signaling pathways , resulting in altered protein expression . Arundic acid [ ( R ) -(-)-2-propyloctanoic acid , DB05343 ] is a novel agent that inhibits P04271 synthesis in cultured astrocytes . In the rodent ischemia model , this agent was shown to inhibit both the astrocytic overexpression of P04271 and the subsequent activation of signaling pathways in the peri-infarct area . Concurrently , delayed infarct expansion was prevented , and neurologic deficits were promptly ameliorated . The results of subsequent studies suggest that the efficacy of arundic acid is mediated by restoring the activity of astroglial glutamate transporters via enhanced genetic expression . The use of CD31 and collagen IV as vascular markers . A study of 56 vascular lesions . A monoclonal antibody against the endothelial cell adhesion molecule CD31 ( endo- P62158 , P16284 ) was tested on wax embedded tissue of 56 cases of vascular lesions including benign and malignant vascular neoplasms . Additional preparations were stained with antibodies to type IV collagen and to P04275 /Factor VIII-RAG ( vWf ) . Our results prove reliability of CD31 and its superiority to vWf in the labelling of endothelium and its subsets in such lesions , with the exception of lymphangiomas . As markers for basal lamina collagen and endothelium respectively , type IV collagen and CD31 together provide a powerful tool for 1 ) the diagnosis of endothelial neoplasms and 2 ) the definition of endothelial , mural and pericytic or perivascular tissue compartments in vascular lesions of complex architecture . Hence , we use CD31 and type IV collagen in cases of presumed vascular neoplasms , adding other markers to the panel in accordance with the differential diagnosis , as well as in the recognition of compromised endothelia , such as in vascular invasion by various malignant neoplasms . 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 ) Proangiogenesis action of the thyroid hormone analog 3,5-diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2- to 3-fold by either T(4) or T(3) at 10(-8)-10(-7) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5-diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T(3) and T(4) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T(4) and T(3) for a novel cell surface hormone receptor site on integrin alphavbeta3 . The thyroid hormone analogs DITPA , T(4) , and T(4)-agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 /2 signal transduction cascade . Additionally , a specific integrin alphavbeta3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta3 , and are MAPK dependent . The cardiac L-type calcium channel distal carboxy terminus autoinhibition is regulated by calcium . The L-type calcium channel ( LTCC ) provides trigger Ca(2+) for sarcoplasmic reticulum Ca-release , and LTCC function is influenced by interacting proteins including the LTCC distal COOH terminus ( P40126 ) and calmodulin . P40126 is proteolytically cleaved and reassociates with the LTCC complex to regulate calcium channel function . P40126 reduces LTCC barium current ( I(Ba,L) ) in reconstituted channel complexes , yet the contribution of P40126 to LTCC Ca(2+) current ( I(Ca,L) ) in cardiomyocyte systems is unexplored . This study tests the hypothesis that P40126 attenuates cardiomyocyte I(Ca,L) . We measured LTCC current and Ca(2+) transients with P40126 coexpressed in murine cardiomyocytes . We also heterologously coexpressed P40126 and Ca(V)1.2 constructs with truncations corresponding to the predicted proteolytic cleavage site , Ca(V)1.2Δ1801 , and a shorter deletion corresponding to well-studied construct , Ca(V)1.2Δ1733 . P40126 inhibited I(Ba,L) in cardiomyocytes , and in human embryonic kidney ( P29320 ) 293 cells expressing Ca(V)1.2Δ1801 and Ca(V)1.2Δ1733 . Ca(2+)- P62158 relieved P40126 block in cardiomyocytes and P29320 cells . The selective block of I(Ba,L) combined with Ca(2+)- P62158 effects suggested that P40126 -mediated blockade may be relieved under conditions of elevated Ca(2+) . We therefore tested the hypothesis that P40126 block is dynamic , increasing under relatively low Ca(2+) , and show that P40126 reduced diastolic Ca(2+) at low stimulation frequencies but spared high frequency Ca(2+) entry . P40126 reduction of diastolic Ca(2+) and relief of block at high pacing frequencies and under conditions of supraphysiological bath Ca(2+) suggests that a physiological function of P40126 is to increase the dynamic range of Ca(2+) transients in response to elevated pacing frequencies . Our data motivate the new hypothesis that P40126 is a native reverse use-dependent inhibitor of LTCC current . Direct binding of recombinant plasminogen kringle 1-3 to angiogenin inhibits angiogenin-induced angiogenesis in the chick embryo P62158 . P03950 is one of the most potent angiogenesis-inducing proteins . Angiostatin is one of the most potent angiogenesis inhibitors , and it contains the first four kringle domains of plasminogen ( P04264 -4 ) . Recombinant human plasminogen kringle 1-3 ( rK1-3 ) was expressed in Escherichia coli and purified to homogeneity . The binding of t-4-aminomethylcyclohexanecarboxylic acid with the purified kringle 1-3 was determined by changes in intrinsic fluorescence . rK1-3 exhibits comparable ligand-binding properties as native human plasminogen kringle 1-3 . The purified rK1-3 inhibits neovascularization in the chick embryo chorioallantoic membrane ( P62158 ) assay . Interaction of angiogenin with rK1-3 was examined by immunological binding assay and surface plasmon resonance kinetic analysis , and the equilibrium dissociation constants for the complex , Kd , are 0.89 and 0.18 microM , respectively . rK1-3 inhibits angiogenin-induced angiogenesis in the chick embryo P62158 in a concentration-dependent manner . These results indicate that rK1-3 directly binds to angiogenin and thus rK1-3 inhibits the angiogenic activity of angiogenin . 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 . 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 . Clinical pharmacology of albiglutide , a P43220 agonist . DB09043 is a glucagon-like peptide-1 analogue composed of tandem copies of modified human glucagon-like peptide-1 ( 7-36 ) coupled to recombinant human albumin that is approved in adults for the treatment of type 2 diabetes mellitus . After subcutaneous administration , albiglutide is likely primarily absorbed via the lymphatic circulation , with maximum concentrations being reached in 3 to 5 days ; steady-state exposures are achieved following approximately 4 to 5 weeks of once-weekly administration . The elimination half-life of albiglutide is approximately 5 days . Clearance of albiglutide is 67 mL/h with between-subject variability of 34.9 % ; no covariates have been identified that would require dose adjustment of albiglutide . DB09043 lowers the fasting plasma glucose and reduces postprandial glucose excursions . In addition , β-cell secretion is enhanced by albiglutide during hyperglycemia , whereas secretion is suppressed during hypoglycemia ; α-cell response to hypoglycemia is not impaired by albiglutide . DB09043 does not prolong the corrected QT interval but has a modest effect on heart rate in patients with type 2 diabetes mellitus . Dose adjustment is not suggested in patients with renal impairment , but experience in patients with severe renal impairment is very limited , and it is recommended that albiglutide be used with care in such patients due to an increased frequency of diarrhea , nausea , and vomiting . No clinically relevant drug interactions have been observed in clinical trials . TRIAL REGISTRATION : NCT00938158 , NCT01406262 , NCT00537719 , NCT01077505 , NCT01147731 , NCT01147718 , NCT01147692 , NCT00354536 , NCT00394030 , NCT00530309 , NCT01357889 , NCT00518115 , NCT01098461 , NCT01475734 , NCT00849017 , NCT00838916 , NCT00839527 , NCT01098539 . Neutrophil growth factors . A significant advance in the field of neutrophil growth factors has occurred with the commercial availability of pegfilgrastim ( Neulasta , Amgen , Thousand Oaks , CA ) , a new-generation , pegylated filgrastim molecule with a sustained duration of action . Pegylation of filgrastim allows once-per-chemotherapy cycle frequency of administration , in contrast to repeated daily administration of filgrastim . Clinical data from two randomized trials demonstrate equivalence of pegfilgrastim and filgrastim in duration of severe neutropenia and recovery from absolute neutrophil count nadir following myelosuppressive chemotherapy . In addition , secondary endpoint results in both trials suggest an enhanced reduction in the overall incidence of febrile neutropenia with pegfilgrastim . Neutrophil kinetic studies demonstrate steady serum neutrophil levels following pegfilgrastim administration , in contrast to the peak-and-trough neutrophil effects observed following filgrastim administration . P09919 ( DB00099 ) therapy has an antiapoptotic effect on neutrophils , which may be enhanced by continuous serum concentrations of pegfilgrastim . Monocytes possess a Q99062 , and this finding has fueled investigational analysis of the role of G- P04141 as a mediator in the host inflammatory response to foreign pathogens . The data demonstrate that depending on the timing of administration , G- P04141 may function as a proinflammatory mediator or an anti-inflammatory mediator . It is likely that the early , prophylactic administration of pegfilgrastim creates an environment in which an anti-inflammatory response predominates . Additional investigational studies will be necessary to confirm and better define the mechanism for enhanced benefit of pegfilgrastim over filgrastim . The recent biologic findings of the mechanism of G- P04141 therapy reviewed here provide a strong basis from which further research initiatives may be conducted .

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interacts_with DB00072?

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Regulation of P40763 by histone deacetylase-3 in diffuse large B-cell lymphoma : implications for therapy . Diffuse large B-cell lymphoma ( DLBCL ) with an activated B-cell ( DB01048 ) gene-expression profile has been shown to have a poorer prognosis compared with tumors with a germinal center B-cell type . ABC cell lines have constitutive activation of P40763 ; however , the mechanisms regulating P40763 signaling in lymphoma are unknown . In studies of class-I histone deacetylase ( HDAC ) expression , we found overexpression of O15379 in phospho P40763 -positive DLBCL and the O15379 was found to be complexed with P40763 . Inhibition of HDAC activity by DB06603 ( LBH589 ) increased p300-mediated P40763 (Lys685) acetylation with increased nuclear export of P40763 to the cytoplasm . HDAC inhibition abolished P40763 (Tyr705) phosphorylation with minimal effect on P40763 (Ser727) and O60674 tyrosine activity . pSTAT3(Tyr705)-positive DLBCLs were more sensitive to HDAC inhibition with LBH589 compared with pSTAT3(Tyr705)-negative DLBCLs . This cytotoxicity was associated with downregulation of the direct P40763 target Mcl-1 . O15379 knockdown upregulated P40763 (Lys685) acetylation but prevented P40763 (Tyr705) phosphorylation and inhibited survival of pSTAT3-positive DLBCL cells . These studies provide the rationale for targeting P40763 -positive DLBCL tumors with HDAC inhibitors . 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 . 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 . 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 . 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 . 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 . 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 . P21860 as biomarker and therapeutic target in pancreatic cancer : new insights in pertuzumab therapy in preclinical models . The anti- P04626 antibody pertuzumab inhibits P04626 dimerization and affects P04626 / P21860 dimer formation and signaling . As P21860 and its ligand neuregulin are implicated in pancreatic tumorigenesis , we investigated whether P21860 expression could be a predictive biomarker of pertuzumab efficacy in HER2low-expressing pancreatic cancer . We correlated in vitro and in vivo P21860 expression and neuregulin dependency with the inhibitory effect of pertuzumab on cell viability and tumor progression . P21860 knockdown in BxPC-3 cells led to resistance to pertuzumab therapy . DB06366 treatment of P21860 -expressing pancreatic cancer cells increased P21860 at the cell membrane , whereas the anti- P21860 monoclonal antibody 9F7- P03951 down-regulated it . Both antibodies blocked P21860 and AKT phosphorylation and inhibited P04626 / P21860 heterodimerization but affected differently P04626 and P21860 homodimers . The pertuzumab/9F7- P03951 combination enhanced tumor inhibition and the median survival time in mice xenografted with P21860 -expressing pancreatic cancer cells . Finally , P04626 and P21860 were co-expressed in 11 % and P21860 alone in 27 % of the 45 pancreatic ductal adenocarcinomas analyzed by immunohistochemistry . P21860 is essential for pertuzumab efficacy in HER2low-expressing pancreatic cancer and P21860 expression might be a predictive biomarker of pertuzumab efficacy in such cancers . Further studies in clinical samples are required to confirm these findings and the interest of combining anti- P04626 and anti- P21860 therapeutic antibodies . Association of folate with hearing is dependent on the 5,10-methylenetetrahdyrofolate reductase 677C --> T mutation . Vascular disease and its risk factors have been associated with the age-related hearing loss . We examined the association of elevated plasma homocysteine and its determinants with hearing levels . Pure-tone air conduction thresholds in 728 individuals with sensorineural hearing loss were not associated with homocysteine , erythrocyte folate and DB00165 . Low concentrations of serum folate and DB00115 were associated with better hearing . When folate status was below the median , 5,10-methylenetetrahydrofolate reductase ( P42898 ) 677TT homozygotes had similar hearing levels to subjects with a C allele . However , when folate status was above the median , P42898 677TT homozygotes had on an average 5 dB ( p = 0.037 ) and 2.6 dB ( p = 0.021 ) lower P03951 -high and P03951 -low hearing thresholds , respectively , than the subjects with a 677C allele . The relationship between serum folate and hearing thresholds appeared to be dependent on P42898 677 genotype ( CC , r = 0.13 , p = 0.034 ; TT , r = -0.10 , p = 0.291 ) . This supports the hypothesis that a greater one-carbon moiety commitment to de novo synthesis of nucleotides and an increase in formyl-folate derivatives relative to methyl-folate derivatives is protective for hearing . DB00439 potentiates nitric oxide release and enos expression through inhibition of isoprenoids synthesis . Endothelium dysfunction , which is often defined as a decrease in NO bioavailability , is one of the earliest manifestations of endothelium-impaired function disorders , including atherosclerosis . Although improvement in NO bioavailability has been attributed to the lowering of serum cholesterol levels , recent studies suggest that P04035 inhibitors , statins , may have direct effects on NO bioavailability by little known mechanisms that are independent of serum cholesterol levels . The long-term effect of cerivastatin on NO release from endothelial cells was determined by using highly sensitive electrochemical microsensors and was correlated with endothelial NO synthase ( P29474 ) levels . To explore whether changes in isoprenoid synthesis affect NO bioavailability and P29474 expression , human endothelial cells were treated with cerivastatin , L-mevalonate ( MVA ; 1.5 mmol/L ) , geranylgeranylpyrophosphate ( GGPP ; 1 mg/mL ) and farnesylpyrophosphate ( FPP ; 1 mg/mL ) . DB00439 increased spontaneous ( by 53 % +/- 6 ) and an P29474 -stimulated NO release ( by 41 +/- 6 % for calcium ionophore and by 47 +/- 5 % acetylcholine ) as well as P29474 expression ( by 118 +/- 6 % ) in the same concentration-range . DB00439 -dependent increase in both NO release and P29474 expression was revealed after approximately 4 h of exposure reaching the maximum after approximately 10 h . Co-treatment with MVA or GGPP , but not FPP or LDL , reversed the effects of cerivastatin . These findings indicate that the long-term effect of cerivastatin resulting in enhanced NO bioavailabilty in endothelial cell is , at least in part , due to up-regulation of P29474 by blocking isoprenoids synthesis . 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 . Nongenomic mechanisms of endothelial nitric oxide synthase activation by the selective estrogen receptor modulator raloxifene . BACKGROUND : Nontranscriptional signaling through estrogen receptors ( ERs ) is important in the cardiovascular system . In particular , estrogen stimulates endothelial NO synthase ( P29474 ) via the phosphatidylinositol 3-kinase ( PI3K ) pathway . The selective estrogen receptor modulator ( SERM ) raloxifene is effective for the treatment of postmenopausal osteoporosis , but its ability to activate P29474 via PI3K is unknown . METHODS AND RESULTS : Human umbilical vein endothelial cells were cultured in estrogen-deprived , phenol red-free medium . Raloxifene stimulated P29474 in a concentration- and time-dependent manner . Activation of P29474 by raloxifene was blocked by the PI3K inhibitor wortmannin and by the ER antagonist DB00947 but not by transcriptional or translational inhibitors . Coimmunoprecipitation studies demonstrated that , in a ligand-dependent manner , raloxifene increased ERalpha-associated p85alpha , P42336 , and PI3K activity . This correlated temporally with increases in the serine and threonine phosphorylation and activation of protein kinase Akt . CONCLUSIONS : Our findings indicate that nongenomic ER signaling triggered by a SERM leads to a rapid activation of NO synthesis in human endothelial cells . The ability of raloxifene to facilitate ERalpha-PI3K interaction may provide additional insight into the structure-function relationship of specific SERMs , which promote the nontranscriptional effects of ER . Regulation of pancreatic cancer by neuropsychological stress responses : a novel target for intervention . Pancreatic cancer has a poor prognosis and is associated with high levels of psychological stress that may adversely affect clinical outcomes . However , the potential influence of neuropsychological factors on pancreatic cancer has not been investigated to date . Using a mouse model of social stress , we have tested the hypothesis that psychological stress promotes the progression of pancreatic cancer xenografts via neurotransmitter-induced activation of multiple pathways and that the inhibitory neurotransmitter γ-aminobutiric acid ( GABA ) inhibits these responses . Sytemic and xenograft levels of noradrenalin , adrenalin , GABA , cortisol , vascular endothelial growth factor ( P15692 ) and cyclic adenosine 3' , 5'-monophosphate ( DB02527 ) were measured by immunoassays . Xenograft expression of nicotinic acetylcholine receptors ( nAChRs ) α3 , α4 , α5 , α6 and α7 and β-adrenergic receptors 1 and 2 were assessed by real-time PCR and western blots . Expression of glutamate decarboxylases Q05329 and Q99259 and phosphorylated and unphosphorylated signaling proteins of relevance to pancreatic cancer were determined in tumor tissue by western blots . Psychological stress significantly promoted xenograft growth and increased systemic and tumor levels of noradrenalin , adrenalin , cortisol , P15692 and DB02527 while GABA and Q99259 were suppressed . Stress upregulated nAChR proteins but not RNAs and induced phosphorylated P29323 , CREB , Src and AKT in xenografts . Reduction of DB02527 by treatment with GABA prevented tumor progression and activation of signaling proteins . Our findings suggest that neurotransmitter responses to psychological stress negatively impact clinical outcomes of pancreatic cancer via the activation of multiple pathways and that replacement of the suppressed inhibitory neurotransmitter GABA prevents these effects . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . DB01373 -sensing receptor A986S polymorphism in human rectal cancer . BACKGROUND AND AIMS : In vivo and in vitro experiments show the protective role of calcium ions ( Ca2+ ) against colorectal cancer . The calcium-sensing receptor ( P41180 ) detects extracellular Ca2+ concentration . An association between the P41180 A986S polymorphism and serum calcium in healthy adults has been reported . Subjects with AA genotype had lower serum concentrations of Ca2+ than other genotypes . The expression of erbB-2 , epidermal growth factor receptor ( P00533 ) , p53 , and ras in colorectal cancer has been suggested to have diagnostic and prognostic significance . PATIENTS AND METHODS : We investigated the relationship between the P41180 A986S polymorphism and the expression of erbB-2 , P00533 , p53 , and ras as well as the UICC stage in 56 patients with rectal cancer . RESULTS : The occurrence of the genotype AA was not different in cancer patients and in 112 controls . In the presence of the coexpression of major oncogenes , patients with genotype AA were in significantly higher UICC stages than in the case of AS genotype . During the follow-up period AA genotype showed a tendency for poor prognosis . CONCLUSIONS : Our observation raises the possibility that genetic alterations of P41180 influence the pathogenesis of rectal cancer . Targetting esophageal and gastric cancers with monoclonal antibodies . Target therapies and notably monoclonal antibodies are currently being considered for esophageal , gastric , and gastroesophageal junction cancers . P00533 was found to be overexpressed in 60-86 % of gastric or gastroesophageal tumors and in 50-70 % of esophageal cancers . Cetuximab was shown to be a radiosensitizing agent in the treatment of ENT neoplasia . These results led to several phase II encouraging therapeutic trials evaluating the combination of cetuximab with radiochemotherapy in locally advanced esophageal cancers . Numerous encouraging phase II trials evaluating cetuximab combined with chemotherapy in patients with gastric adenocarcinoma or gastroesophageal junction cancer were reported . These promising results are still to be confirmed by the ongoing phase III trials . Several studies reported P04626 overexpression in gastric cancer ( 7-34 % ) , which appeared to be associated with poorer prognosis . DB00072 is a monoclonal antibody directed against the extracellular P04626 domain . The international phase III trial known as ToGA ( DB00072 for Gastric Cancer ) aimed to determine the clinical efficacy and acceptable toxicity profile of trastuzumab in combination with first-line chemotherapy in P04626 -overexpressing gastric or gastroesophageal cancer . Angiogenesis is an essential step in the initial phase of tumorigenesis , and it is normally absent from healthy tissues except for particular physiological situations , such as wound healing . P15692 plays a role in endothelial growth and angiogenesis . DB00112 , a humanized monoclonal anti- P15692 antibody , is currently being studied for gastric cancer . The phase III AVAGAST study , evaluating bevacizumab in association with chemotherapy in advanced gastric adenocarcinoma , did not achieve its primary aim of improved OS in bevacizumab-treated patients . cDNA microarray reveals signaling pathways involved in hormones expression of human pituitary . Pituitary , a master gland of neuroendocrine system , secretes hormones that orchestrate many physiological processes , under the regulation of multiple signaling pathways . To investigate the genes involved in hormones expression of human pituitary , homemade cDNA microarray containing 14,800 human genes/ESTs were used to profile the gene expression in both fetal and adult pituitaries . Seven hundred and twelve known genes changed over 2-fold between the both tissues . Of which , 23 genes were changed with hormones expression in aging were confirmed by RT-PCR , not only the known regulators such as Pit1 , P43694 , P11474 , GABA-A , and EMK , but also LOC55884 , P51452 , Q9H307 , and O43598 , which had not been reported to be involved in the hormones expression . Correspondingly , the mRNAs of GH , PRL , P01189 , P01222 , DB00094 -beta , and LH-beta , was increased as much as 6- to 20-fold in adult pituitary than those in fetal pituitary , by real-time quantitative RT-PCR assay . In addition , the mRNAs of signaling pathways , such as DB02527 -PKA-CREB , PI3K-Akt , and PKA- P29323 were further investigated . Of them , it was only DB02527 -PKA-CREB pathway , but not PI3K-Akt and PKA- P29323 have the same expressing pattern as hormones . It suggested that cDNA microarray is highly advantages to profile the differential expressed genes that were involved in hormones expression of human pituitary , but it might ignore some responding proteins regulated posttranscriptionally . 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 . 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 . Antibody-dependent cell cytotoxicity to breast cancer targets despite inhibitory P55040 signaling . BACKGROUND : Natural killer ( NK ) cells express killer immunoglobulin-like ( P55040 ) inhibitory receptors , which recognize certain HLA class I molecules ( P55040 ligands ) , and stimulatory receptors such as FcgammaRIII . The purpose of this study was to test the possible influence of inhibitory P55040 signaling on antibody-dependent cell cytotoxicity ( ADCC ) mediated by allogeneic NK cells against breast cancer targets . MATERIALS AND METHODS : The cytotoxic activity of volunteer donor NK cells against the cell lines SKBR-3 , T47D and MCF-7 , which have high , low and no P04626 gene amplification , respectively , were studied . Both cell lines and donors were assigned to the C1 or P06681 superfamily , defined by the structure of the HLA-Cw molecule . RESULTS : It was found that ADCC mediated by allogeneic NK cells occurred despite combinations of NK cells and breast cancer targets predicted to trigger inhibitory P55040 signaling . CONCLUSION : We suggest that adoptive immunotherapy with allogeneic NK cells and trastuzumab may be an effective combination against breast cancer targets . Analog of somatostatin vapreotide exhibits biological effects in vitro via interaction with neurokinin-1 receptor . OBJECTIVES : DB04894 , a synthetic analog of somatostatin , has analgesic activity most likely mediated through the blockade of neurokinin-1 receptor ( P25103 ) , the DB05875 ( SP ) -preferring receptor . The ability of vapreotide to interfere with other biological effects of SP has yet to be investigated . METHODS : We studied the ability of vapreotide to antagonize P25103 in three different cell types : immortalized U373MG human astrocytoma cells , human monocyte-derived macrophages ( MDM ) and a human embryonic kidney cell line , HEK293 . Both U373MG and MDM express endogenous P25103 while HEK293 cells , which normally do not express P25103 , are stably transformed to express human P25103 ( HEK293- P25103 ) . RESULTS : DB04894 attenuates SP-triggered intracellular calcium increases and nuclear factor-κB activation in a dose-dependent manner . DB04894 also inhibits SP-induced interleukin-8 and monocyte chemotactic protein-1 production in HEK293- P25103 and U373MG cell lines . DB04894 inhibits HIV-1 infection of human MDM in vitro , an effect that is reversible by SP pretreatment . CONCLUSIONS : Our findings indicate that vapreotide has P25103 antagonist activity and may have a potential application as a therapeutic intervention in HIV-1 infection . Substance P autocrine signaling contributes to persistent P04626 activation that drives malignant progression and drug resistance in breast cancer . P00533 receptor transmodulation by heterologous G-protein-coupled receptors ( GPCR ) generates functional diversity in signal transduction . Tachykinins are neuropeptides and proinflammatory cytokines that promote cell survival and cancer progression by activating several GPCRs . In this work , we found that the pain-associated tachykinin Substance P ( SP ) contributes to persistent transmodulation of the P00533 receptors , P00533 and P04626 , in breast cancer , acting to enhance malignancy and therapeutic resistance . SP and its high-affinity receptor P25103 were highly expressed in P04626 (+) primary breast tumors ( relative to the luminal and triple-negative subtypes ) and were overall correlated with poor prognosis factors . In breast cancer cell lines and primary cultures derived from breast cancer samples , we found that SP could activate P04626 . Conversely , RNA interference-mediated attenuation of P25103 , or its chemical inhibition , or suppression of overall GPCR-mediated signaling , all strongly decreased steady-state expression of P00533 and P04626 , establishing that their basal activity relied upon transdirectional activation by GPCR . Thus , SP exposure affected cellular responses to anti- P00533 therapies . Our work reveals an important oncogenic cooperation between P25103 and P04626 , thereby adding a novel link between inflammation and cancer progression that may be targetable by SP antagonists that have been clinically explored . DB00072 inhibits the growth of human gastric cancer cell lines with P04626 amplification synergistically with cisplatin . P04626 has been found to be amplified in 10-20 % of gastric cancers , and is correlated with poor outcome . The aims of this study were to recognize P04626 amplification in gastric cancer cell lines via fluorescence in situ hybridization and to evaluate the growth inhibitory effect of trastuzumab in P04626 -amplified cell lines . To elucidate the mechanism of the growth inhibition , we performed cell cycle analysis and immunoblotting of downstream molecules . We also conducted drug interaction studies of trastuzumab with other chemotherapeutic agents . P04626 amplification was newly identified only in SNU-216 cells , and trastuzumab moderately inhibited the growth of SNU-216 cells and positive controls . DB00072 -mediated P55008 arrest occurred with increased expression of p27( P46527 ) and decreased cyclins . Phosphorylation of P04626 and downstream molecules , P40763 , AKT , and P29323 , was also inhibited by trastuzumab . Treatment of SNU-216 cells with trastuzumab plus cisplatin resulted in a synergistic inhibitory effect , whereas treatment of SNU-216 cells with trastuzumab plus DB00544 , or trastuzumab plus oxaliplatin produced an additive effect . These results suggest that trastuzumab combined with chemotherapeutic agents can be active against gastric cancer with P04626 amplification .

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interacts_with DB04844?

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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 . DB00092 : a safety profile . DB00092 is a selective immunomodulating , antipsoriatic drug that blocks the LFA-3/ P06729 interaction necessary for the activation and proliferation of memory effector T cells by binding to P06729 expressed on the T cell surface . Because the P01730 + count is reduced by alefacept , it is recommended that this count be monitored on a regular basis to ensure that it does not drop below 250 cells/mul . Few side effects have been related to the use of alefacept that differ from placebo even when P01730 + counts drop below 250 cells/microl . The side effects that have been reported are minor and include : headache , nasopharyngitis , rhinitis , influenza , upper respiratory tract infections , pruritus , arthralgias , fatigue , nausea , accidental injury and increases in liver enzymes . Serious infections and malignancies do not appear linked to the use of alefacept . The percentage of patients who developed antibodies against alefacept is very low . DB00092 is a very safe biological therapy for moderate-to-severe chronic plaque psoriasis with few side effects reported . The utility of checking P01730 counts while administering alefacept for 12 weeks appears minimal . DB01954 attenuates acute oligodendrocyte death in the adult rat ventrolateral funiculus following contusive cervical spinal cord injury . DB01954 , an inhibitor of phosphodiesterase 4 ( DB05876 ) proteins that hydrolyze DB02527 , increases axonal regeneration following spinal cord injury ( SCI ) . Recent evidence indicate that rolipram also protects against a multitude of apoptotic signals , many of which are implicated in secondary cell death post-SCI . In the present study , we used immunohistochemistry and morphometry to determine potential spinal cord targets of rolipram and to test its protective potential in rats undergoing cervical spinal cord contusive injury . We found that 3 DB05876 subtypes ( P27815 , B , D ) were expressed by spinal cord oligodendrocytes . OX-42 immunopositive microglia only expressed the Q07343 subtype . Oligodendrocyte somata were quantified within the cervical ventrolateral funiculus , a white matter region critical for locomotion , at varying time points after SCI in rats receiving rolipram or vehicle treatments . We show that rolipram significantly attenuated oligodendrocyte death at 24 h post-SCI continuing through 72 h , the longest time point examined . These results demonstrate for the first time that spinal cord glial cells express DB05876 subtypes and that the DB05876 inhibitor rolipram protects oligodendrocytes from secondary cell death following contusive SCI . They also indicate that further investigations into neuroprotection and axonal regeneration with rolipram are warranted for treating SCI . Effects of noradrenergic lesions on the development of rolipram-sensitive , low-K(m) , cyclic AMP specific phosphodiesterase in rat brain . DB01954 -sensitive , low-K(m)80 % loss of norepinephrine in cerebral cortex ) without affecting dopaminergic systems . The lesions resulted in temporary reduction of DB05876 activity in cerebral cortex , cerebellum and brainstem . Lesions in the adult rats , on the other hand , did not alter DB05876 activity . Decreased DB05876 activity by neonatal noradrenergic lesions was due to a decrease in the V(max) of DB02527 hydrolysis by DB05876 , and not a change in the K(m) values . Immunoblot analysis showed that decreased DB05876 activity in cerebellum was associated with reduced expression of PDE4A5 , PDE4A1 , and several Q07343 variants . No change in the expression of any DB05876 subtype in cerebral cortex was detected with the antibodies used in this study . Neither the permanent loss of noradrenergic innervation in cerebral cortex , nor the permanent noradrenergic hyperinnervation in brainstem was accompanied by any permanent change in DB05876 activity . Decreasing DB05876 activity early after neonatal noradrenergic lesions might be important in maintaining constant concentrations of DB02527 , which is critical for the cellular proliferation and differentiation that is active during this period . DB00714 -induced aggressiveness and [3H]citalopram binding after antidepressant treatment in rats . The effects of acute and repeated administration of antidepressive drugs on apomorphine-induced aggressive behavior and [3H]citalopram binding were studied . In acute behavioral experiments with apomorphine pretreated ( 1.0 mg/kg , once daily ) animals , desipramine ( 10 mg/kg ) and clomipramine ( 10 mg/kg ) enhanced , buspirone ( 2.5 and 5.0 mg/kg ) completely blocked , but fluoxetine , amitriptyline , imipramine ( 10 mg/kg ) , and citalopram ( 10 and 20 mg/kg ) had no effect on the intensity of aggressive behavior . Repeated concomitant apomorphine ( 1.0 mg/kg ) and citalopram ( 10 mg/kg ) administration reduced the affinity ( Kd ) of the 5-HT transporter binding sites in three brain regions . This finding was confirmed by an additional experiment as the effect of citalopram treatment . Repeated apomorphine ( 1.0 mg/kg ) or apomorphine ( 1.0 mg/kg ) plus desipramine ( 10 mg/kg ) treatment had no unidirectional effect on Kd , the maximal number of apparent binding sties ( Bmax ) was unchanged in all experiments . Our study indicates that the 5-HT reuptake blockade has no major influence on the apomorphine-induced aggressive behavior , but the P08908 receptor subtype may be involved in the mediation of the aggressive behavior in this paradigm . 17beta-estradiol induces Akt-1 through estrogen receptor-beta in the frog ( Rana esculenta ) male germ cells . Several lines of evidence support the key role of estrogens in male fertility . Here , we investigate the regulation of the serine/threonine kinase Akt-1 in the frog ( Rana esculenta ) testis during the annual sexual cycle and , whether 17beta-estradiol ( E2 ) exerts a role in the Akt-1 activity . Akt-1 has been shown to be the mediator of growth factor-dependent cell proliferation , survival , and metabolism in a variety of cell types . First , we demonstrate by immunohistochemistry , the presence of estrogen receptor-beta ( ERbeta ) , and Akt-1 in the spermatogonia ( Q9HBG6 ) , spermatocytes ( Q969E3 ) , and spermatids ( P21549 ) . Western-blot analysis revealed that ERbeta isoform ( molecular weight 55 kDa ) was highly expressed in May ( reproductive period ) with respect to January and November ( winter stasis ) ; in parallel , Akt-1 ( molecular weight 60 kDa ) is highly phosphorylated ( DB00133 -473 ) during the period of active spermatogenesis ( May ) compared with the winter stasis ( January and November ) . In addition , in vitro experiments demonstrate that E2 treatment induces the activation of Akt-1 , and this effect is counteracted by the anti-estrogen ICI 182-780 . In conclusion , our data show that E2 induces Akt-1 phosphorylation ( DB00133 -473 ) possibly via ERbeta in frog ( R. esculenta ) male germ cells . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . 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 . Transforming growth factor-alpha directly augments histidine decarboxylase and vesicular monoamine transporter 2 production in rat enterochromaffin-like cells . For the production and vesicle storage of histamine , Enterochromaffin-like ( ECL ) cells express histidine decarboxylase ( HDC ) and vesicular monoamine transporter 2 ( Q05940 ) . Although HDC and Q05940 show dynamic changes during gastric ulcer healing , the control system of their expression has not been fully investigated . In the present study , we investigated the effect of transforming growth factor-alpha ( TGF-alpha ) and proinflammatory cytokines on HDC and Q05940 expression in rat ECL cells . Time course changes in the expression of TGF-alpha during the healing of acetic acid-induced ulcers were studied . P01133 receptor ( P00533 ) expression was also examined in ECL cells , whereas the direct effects of TGF-alpha and proinflammatory cytokines on HDC and Q05940 expression in ECL cells were investigated using in vivo and in vitro models . During the process of ulcer healing , expression of TGF-alpha mRNA was markedly augmented . Furthermore , P00533 was identified in isolated ECL cells . TGF-alpha stimulated HDC and Q05940 mRNA expression and protein production and also increased histamine release from ECL cells . Selective P00533 tyrosine kinase inhibitor tyrphostin AG1478 almost completely inhibited HDC and Q05940 gene expression induced by TGF-alpha in vivo and in vitro . During gastric mucosal injury , TGF-alpha was found to stimulate ECL cell functions by increasing HDC and Q05940 expression . Parkinson 's disease-associated alpha-synuclein is a calmodulin substrate . P37840 is a neuronal protein thought to be central in the pathogenesis of Parkinson 's disease ( PD ) because it comprises the fibrillar core of Lewy bodies , one of the histologically defining lesions of PD , and because mutations in alpha-synuclein cause autosomal dominant PD . Although its physiologic role is uncertain , alpha-synuclein is a synaptic protein that may contribute to plasticity . We produced synuclein with incorporated photoprobes to identify and purify novel synuclein-interacting proteins both to begin to clarify the physiology of synuclein and to identify factors that may regulate synuclein conformation . We detected several cross-links and purified and identified one as calmodulin ( P62158 ) . P62158 binds to both wild type and PD-associated mutant alpha-synucleins in a calcium-dependent manner . We further demonstrate that P62158 and alpha-synuclein interact in intact cells in a calcium-dependent manner and that activated P62158 accelerates the formation of synuclein fibrils in vitro . We hypothesize that the known calcium control of synuclein function is mediated through P62158 interaction and that P62158 potentially alters synuclein conformation . Explorative immunohistochemical study to evaluate the addition of a topical corticosteroid in the early phase of alefacept treatment for psoriasis . The aim of this study was to explore the additional effect of betamethasone dipropionate cream in the early phase of an intramuscular ( IM ) alefacept course , on plaque severity and on modulating T-cell subsets , cells expressing NK-receptors , epidermal proliferation and keratinocyte differentiation in lesional psoriatic skin . Therefore , sixteen patients with moderate-to-severe chronic plaque psoriasis received 15 mg alefacept IM for 12 weeks , followed by a 12-week follow-up period . The first 4 weeks , patients were randomized 1:1 to either betamethasone dipropionate , or the vehicle cream , once daily . Plaque severity ( SUM ) was assessed and serial biopsies were immunohistochemically stained for T-cell subsets ( CD3 , P01730 , CD8 , CD45RO , CD45RA , P06729 , CD25 , Q9Y5U5 ) , cells expressing NK-receptors ( Q13241 and CD161 ) , epidermal proliferation ( Ki67 ) and differentiation ( P13645 ) , which were quantified using manual and digital image analysis . DB00092 monotherapy resulted in statistically significant improvement in plaque severity . Subsequently , immunohistochemical assessments on T-cell subsets , epidermal proliferation ( Ki67 ) and keratinization ( P13645 ) revealed marked time-related improvements with respect to the mentioned parameters , without significant differences between both treatment regimens . DB00092 monotherapy induces improvement of plaque severity , which is accompanied by a reduction in activated ( P06729 + , CD25+ , CD45RO+ ) dermal P01730 + and activated epidermal CD8+ T cells , epidermal proliferation and differentiation . Once daily treatment with betamethasone dipropionate cream during the first 4 weeks of an intramuscular alefacept course did not provide substantial additional clinical and immunohistochemical improvement . 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 . Diabetes mellitus in cancer patients treated with combination interleukin 2 and alpha-interferon . Diabetes mellitus is thought to be an autoimmune disease caused by destruction of beta cells in pancreatic islets . P01308 resistance in the peripheral tissues may also play a role . Both interleukin 2 ( P60568 ) and alpha interferon can enhance immune function by stimulating formation of cytolytic T cells and/or antigen expression on both normal and tumor cells . This report describes three patients with advanced malignancy who were treated with combination P60568 and alpha interferon who had the onset or worsening of diabetes mellitus . One patient died as a result . There is evidence that interferon can increase insulin resistance and it is likely that both agents can initiate or enhance an ongoing autoimmune process . Physicians using this combination of drugs should be aware of this potential serious toxicity . Estradiol-induced mitogen-activated protein kinase ( extracellular signal-regulated kinase 1 and 2 ) activity in the frog ( Rana esculenta ) testis . Several lines of evidence support a key role of estradiol-17beta ( E(2) ) in male fertility . We have used a non-mammalian vertebrate model , the frog Rana esculenta , to investigate the regulation of extracellular signal-regulated kinase 1 and 2 ( P27361 /2 ) activity in the testis during the annual sexual cycle and to study whether E ( 2 ) exerts a role in spermatogenesis through the regulation of P27361 /2 activity . P27361 /2 proteins are present in the cytoplasm and nucleus of the primary and secondary spermatogonia ( Q9HBG6 ) , and in the nucleus of primary spermatocytes . The annual E(2) profile shows a progressive increase during active spermatogenesis with a peak in the month of June . In parallel , P27361 /2 are highly phosphorylated during the period of active spermatogenesis ( from April to July ) compared with the regressive period ( September/October ) and winter stasis ( from November to March ) . E(2) treatment induces the proliferation of primary Q9HBG6 , possibly via the activation of P27361 /2 , and this effect is counteracted by the anti-estrogen ICI 182-780 . Penile erection and yawning induced by P28335 receptor agonists in male rats : relationship with dopaminergic and oxytocinergic transmission . 1-(3-Chlorophenyl)piperazine ( m-CPP ) ( 0.1-4 mg/kg s.c. ) and N-(3-trifluoromethylphenyl)-piperazine ( TFMPP ) ( 0.5-4 mg/kg s.c. ) , P28335 receptor agonists , but not 8-hydroxy-dipropylamino-tetralin ( 8-OH-DPAT ) ( 0.1 and 0.2 mg/kg s.c. ) , a P08908 receptor agonist , induced penile erection and yawning with a U-inverted dose-response curve in male rats . The maximal effect was found with 0.5 mg/kg s.c. of m-CPP and with 1 mg/kg s.c. of TFMPP . The m-CPP ( 0.5 mg/kg s.c. ) and TFMPP ( 1 mg/kg s.c. ) responses were prevented by mianserin ( 0.2 mg/kg s.c. ) and by ritanserin ( 1 mg/kg s.c. ) given 15 min before m-CPP and TFMPP . In contrast , m-CPP- or TFMPP-induced penile erection and yawning were not antagonized by haloperidol ( 0.1 mg/kg s.c. ) or by [d(CH2)5Tyr(Me)2,Orn8]vasotocin ( 5 micrograms i.c.v. ) . DB00714 - and oxytocin-induced penile erection , but not yawning , was also antagonized by mianserin and less effectively by ritanserin . The results suggest that P28335 receptor agonist-induced penile erection and yawning are not mediated by increased dopaminergic and/or oxytocinergic transmission , and raise the possibility that a neuronal dopamine-oxytocin-5-HT link is involved in the control of penile erection and not necessarily of yawning in male rats . DB09559 , a fully human IgG1 mAb directed against the P00533 for the potential treatment of cancer . DB09559 ( DB05774 ) , under development by ImClone Systems in collaboration with Bristol-Myers Squibb , is a fully human IgG1 mAb targeting the epidermal growth factor receptor ( P00533 ) , for the potential intravenous treatment of cancer , in particular NSCLC . In vitro studies demonstrate that necitumumab inhibits downstream targets in the P00533 pathway ( eg , MAPK ) , which are important for cellular proliferation , differentiation , invasion and metastasis . Furthermore , because necitumumab is an IgG1 construct , it has the potential to induce antibody-dependent cell-mediated cytotoxicity against tumor cells . Preclinical studies indicated that the antitumor activity of necitumumab is either comparable with or superior to that of ImClone 's chimeric anti- P00533 mAb cetuximab . In a phase I clinical trial in patients with advanced solid malignancies , necitumumab displayed nonlinear pharmacokinetic behavior . The toxicity profile of necitumumab is acceptable , with skin toxicity being the most frequently reported adverse event in the phase I and II clinical trials conducted to date . Preliminary data from a phase II clinical trial of necitumumab in combination with chemotherapy for the first-line treatment of advanced colon cancer are promising . Success in the ongoing phase III clinical trials in patients with advanced NSCLC would lead to necitumumab becoming a valuable addition to future therapeutic strategies in oncology . L-454,560 , a potent and selective DB05876 inhibitor with in vivo efficacy in animal models of asthma and cognition . Type 4 phosphodiesterases ( DB05876 ) inhibitors are emerging therapeutics in the treatment of a number of chronic disorders including asthma , chronic obstructive pulmonary disease ( P48444 ) and cognitive disorders . This study delineates the preclinical profile of L-454,560 , which is a potent , competitive and preferential inhibitor of P27815 , 4B , and 4D with IC50 values of 1.6 , 0.5 and 1.2 nM , respectively . In contrast to the exclusive binding of cilomilast and the preferential binding of roflumilast to the DB05876 holoenzyme state ( Mg2+-bound form ) , L-454,560 binds to both the apo-(Mg2+-free) and holoenzyme states of DB05876 . The intrinsic enzyme potency for DB05876 inhibition by L-454,560 also results in an effective blockade of LPS-induced TNFalpha formation in whole blood ( IC50 = 161 nM ) and is comparable to the human whole blood potency of roflumilast . The cytokine profile of inhibition of L-454,560 is mainly a Th1 profile with significant inhibition of IFNgamma and no detectable inhibition of P35225 formation up to 1 microM . L-454,560 was also found to be efficacious in two models of airway hyper-reactivity , the ovalbumin ( OVA ) sensitized and challenged guinea pig and the ascaris sensitized sheep model . Furthermore , L-454560 was also effective in improving performance in the delayed matching to position ( DMTP ) version of the Morris watermaze , at a dose removed from that associated with potential emesis . Therefore , L-454,560 is a novel DB05876 inhibitor with an overall in vivo efficacy profile at least comparable to roflumilast and clearly superior to cilomilast . P63244 recruits P40763 specifically to insulin and insulin-like growth factor 1 receptors for activation , which is important for regulating anchorage-independent growth . Current understanding of the activation of STATs is through binding between the SH2 domain of STATs and phosphotyrosine of tyrosine kinases . Here we demonstrate a novel role of P63244 as an adaptor for insulin and insulin-like growth factor 1 receptor ( IGF-1R ) -mediated P40763 activation specifically . Intracellular association of P63244 via its N-terminal WD domains 1 to 4 ( WD1-4 ) with insulin receptor ( IR ) /IGF-1R is augmented upon respective ligand stimulation , whereas association with P40763 is constitutive . Purified P63244 or P63244 WD1-4 associates directly with purified IR , IGF-1R , and P40763 in vitro . P01308 induces multiprotein complex formation of P63244 , IR , and P40763 . Overexpression or downregulation of P63244 greatly enhances or decreases , respectively , IR/IGF-1R-mediated activation of P40763 and its target gene expression . Site-specific mutants of IR and IGF-1R impaired in P63244 binding are ineffective in mediating recruitment and activation of P40763 as well as in insulin- or DB01277 -induced protection of cells from anoikis . P63244 -mediated P40763 activation is important for insulin and DB01277 -induced anchorage-independent growth in certain ovarian cancer cells . We conclude that P63244 mediates recruitment of P40763 to IR and IGF-1R specifically for activation , suggesting a general paradigm for the need of an adaptor in mediating activation of STATs by receptor protein tyrosine kinases . [ 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 . Intermediate phenotype analysis of patients , unaffected siblings , and healthy controls identifies Q05940 as a candidate gene for psychotic disorder and neurocognition . Psychotic disorders are associated with neurocognitive alterations that aggregate in unaffected family members , suggesting that genetic vulnerability to psychotic disorder impacts neurocognition . The aim of the present study was to investigate whether selected schizophrenia candidate single nucleotide polymorphisms ( SNPs ) are associated with ( 1 ) neurocognitive functioning across populations at different genetic risk for psychosis ( 2 ) and psychotic disorder . The association between 152 SNPs in 43 candidate genes and a composite measure of neurocognitive functioning was examined in 718 patients with psychotic disorder . Follow-up analyses were carried out in 750 unaffected siblings and 389 healthy comparison subjects . In the patients , 13 associations between SNPs and cognitive functioning were significant at P < .05 , situated in P21728 , P35462 , Q01959 , P23560 , P09038 , Q05940 , Q13451 , and Q9UBC3 . Follow-up of these SNPs revealed a significant and directionally similar association for Q05940 ( alternatively Q05940 ) rs363227 in siblings ( B = -0.13 , P = .04 ) and a trend association in control subjects ( B = -0.10 , P = .12 ) . This association was accompanied by a significantly increased risk for psychotic disorder associated with the T allele ( linear OR = 1.51 , 95 % CI 1.10-2.07 , P = .01 ) , which was reduced when covarying for cognitive performance ( OR = 1.29 , 95 % CI 0.92-1.81 , P = .14 ) , suggesting mediation . Genetic variation in Q05940 may be linked to alterations in cognitive functioning underlying psychotic disorder , possibly through altered transport of monoamines into synaptic vesicles . Scaling up antiretroviral treatment services in Karnataka , India : impact on P01730 counts of HIV-infected people . SETTING : Twelve antiretroviral treatment centres under National AIDS Control Programme ( P37840 ) , Karnataka State , India . OBJECTIVE : For the period 2004-2011 , to describe the trends in the numbers of people living with HIV ( PLHIV ) registered for care and their median baseline P01730 counts , disaggregated by age and sex . DESIGN : Descriptive study involving analysis of routinely captured data ( year of registration , age , sex , baseline P01730 count ) under P37840 . RESULTS : 34,882 ( 97 % of total eligible ) PLHIV were included in analysis . The number registered for care has increased by over 12 times during 2004-11 ; with increasing numbers among females . The median baseline P01730 cell count rose from 125 in 2004 to 235 in 2011 -- the increase was greater among females as compared to males . However , about two-thirds still presented at P01730 cell counts less than 350 . CONCLUSION : We found an increasing trend of median P01730 counts among PLHIV presenting to O00253 centres in Karnataka , an indicator of enhanced and early access to HIV care . Equal proportion of females and higher baseline P01730 counts among them allays any fear of differential access by gender . Despite this relative success , a substantial proportion still presented at low P01730 cell counts indicating possibly delayed HIV diagnosis and delayed linkage to HIV care . Universal HIV testing at health care facilities and strengthening early access to care are required to bridge the gap . P01308 augments gonadotropin-releasing hormone induction of translation in LbetaT2 cells . The integrated signaling of insulin and gonadotropin-releasing hormone in the pituitary gonadotropes may have a profound bearing on reproductive function , although the cross-receptor signaling mechanisms are unclear . We demonstrate that the insulin receptor is constitutively localized to non-caveolar lipid raft microdomains in the pituitary gonadotrope cell line LbetaT2 . The localization to rafts is consistent with similar localization of the P30968 . P06213 phosphorylation occurs in raft domains and activates the downstream signaling targets P01308 Receptor Substrate1 and Akt/Protein Kinase B . Although insulin alone does not strongly activate the extracellular signal-regulated kinase second messenger cascade , co-stimulation potentiates the phosphorylation of the extracellular signal-regulated kinase by gonadotropin-releasing hormone . The co-stimulatory effect of insulin and gonadotropin-releasing hormone is also evident in increased activation of cap-dependent translation . In contrast , co-stimulation attenuates Akt/Protein Kinase B activation . Our results show that both gonadotropin-releasing hormone and insulin are capable of mutually altering their respective regulatory signaling cascades . We suggest that this provides a mechanism to integrate neuropeptide and energy homeostatic signals to modulate reproductive function . 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 . 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 . Activation of the unfolded protein response pathway causes ceramide accumulation in yeast and P01308 -1E insulinoma cells . Sphingolipids are not only important components of membranes but also have functions in protein trafficking and intracellular signaling . The O15269 gene encodes a subunit of the serine palmitoyltransferase , which is responsible for the first step of sphingolipid synthesis . Here , we show that activation of the unfolded protein response ( UPR ) can restore normal ceramide levels and viability in yeast cells with a conditional defect in O15269 . Dependence on UPR was demonstrated by showing the HAC1-dependence of the suppression . A similar induction of ceramides by UPR seems to take place in mammalian cells . In rat pancreatic P01308 -1E cells , UPR activation induces the transcription of the Q6ZMG9 gene , which encodes a ceramide synthase . This correlates with the specific accumulation of ceramide with a C16 fatty acyl chain upon UPR activation . Therefore , our study reveals a novel connection between UPR induction and ceramide synthesis that seems to be conserved between yeast and mammalian cells . 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 . Emerging targets of the P30968 : novel interactions with Wnt signalling mediators . The diversity of signalling pathways that gonadotropin-releasing hormone ( DB00644 ) receptors can target has been extended markedly in recent years , ranging from classical heterotrimeric G-protein-coupled second-messenger mobilisation to MAPK cascade activation and epidermal growth factor receptor transactivation . More recently , the targeting of non-classical signalling pathways has been demonstrated , including the activation of monomeric G-proteins and integrin/focal adhesion complexes to mediate cytoskeletal remodelling , androgen receptor nuclear translocation , and luteinising hormone beta-subunit gene transcription . These have been further extended to include c-Src-mediated activation of diacylglycerol kinase , and a novel mechanism of cross-talk to prostaglandin receptor signalling . Here , we review these recent advances and highlight several emerging dimensions in P30968 -mediated signal transduction . These include the targeting of mediators of classical insulin signalling , and Wnt signalling pathways . Collectively , this diverse nature of P30968 signalling is suggestive of an important role for DB00644 in a variety of normal physiological and pathophysiological processes in the reproductive system . A novel synthesis of 2-arylpyrrolo[1,2-a]pyrimid-7-ones and their structure-activity relationships as potent P30968 antagonists . In the process of developing P30968 antagonists , a novel base-catalyzed cyclization of compounds 5a-b was discovered , which led to the formation of the 2-aryl pyrrolo[1,2-a]pyrimid-7-one core structures 6a-b . These intermediates were further modified at positions 1 , 2 , 4 and 6 to afford a series of potent DB00644 antagonists with low nanomolar K(i) values . The P63244 signaling scaffold protein selectively interacts with the DB02527 -specific phosphodiesterase PDE4D5 isoform . The WD-repeat protein receptor for activated C-kinase ( P63244 ) was identified by its interaction with the cyclic AMP-specific phosphodiesterase ( DB05876 ) isoform PDE4D5 in a yeast two-hybrid screen . The interaction was confirmed by co-immunoprecipitation of native P63244 and PDE4D5 from COS7 , HEK293 , 3T3-F442A , and SK-N-SH cell lines . The interaction was unaffected by stimulation of the cells with the phorbol ester phorbol 2-myristate 3-acetate . PDE4D5 did not interact with two other WD-repeat proteins , beta'-coatomer protein and Gsbeta , in two-hybrid tests . P63244 did not interact with other Q08499 isoforms or with known P27815 , Q07343 , and Q08493 isoforms . PDE4D5 and P63244 interacted with high affinity ( Ka approximately 7 nM ) [ corrected ] when they were expressed and purified from Escherichia coli , demonstrating that the interaction does not require intermediate proteins . The binding of the E. coli-expressed proteins did not alter the kinetics of DB02527 hydrolysis by PDE4D5 but caused a 3-4-fold change in its sensitivity to inhibition by the DB05876 selective inhibitor rolipram . The subcellular distributions of P63244 and PDE4D5 were extremely similar , with the major amount of both proteins ( 70 % ) in the high speed supernatant ( S2 ) fraction . Analysis of constructs with specific deletions or single amino acid mutations in PDE4D5 demonstrated that a small cluster of amino acids in the unique amino-terminal region of PDE4D5 was necessary for its interaction with P63244 . We suggest that P63244 may act as a scaffold protein to recruit PDE4D5 and other proteins into a signaling complex . DB00133 palmitoyltransferase , a key enzyme of sphingolipid metabolism . The first step in the biosynthesis of sphingolipids is the condensation of serine and palmitoyl DB01992 , a reaction catalyzed by serine palmitoyltransferase ( P21549 ) to produce 3-ketodihydrosphingosine ( Q9H2K8 ) . This review focuses on recent advances in the biochemistry and molecular biology of P21549 . P21549 belongs to a family of pyridoxal 5'-phosphate ( PLP ) -dependent alpha-oxoamine synthases ( POAS ) . Mammalian P21549 is a heterodimer of 53-kDa O15269 and 63-kDa O15270 subunits , both of which are bound to the endoplasmic reticulum ( ER ) most likely with the type I topology , whereas other members of the POAS family are soluble homodimer enzymes . O15270 appears to be unstable unless it is associated with O15269 . Potent inhibitors of P21549 structurally resemble an intermediate in a probable multistep reaction mechanism for P21549 . Although P21549 is a housekeeping enzyme , its activity is regulated transcriptionally and post-transcriptionally , and its up-regulation is suggested to play a role in apoptosis induced by certain types of stress . Specific missense mutations in the human O15269 gene cause hereditary sensory neuropathy type I , an autosomal dominantly inherited disease , and these mutations confer dominant-negative effects on P21549 activity . Recombinant P17936 inhibits allergic lung inflammation , P15692 production , and vascular leak in a mouse model of asthma . BACKGROUND : Vascular endothelial growth factor ( P15692 ) plays a pro-inflammatory mediator as well as a vascular permeability factor in bronchial asthma . P01308 -like growth factor ( IGF ) -I is also involved in the inflammatory process associated with bronchial asthma and stimulates P15692 expression . The IGF-binding proteins ( IGFBPs ) , especially P17936 , display distinctive properties and can interfere with various biological processes . METHODS : In this study , an ovalbumin ( OVA ) -induced murine model of allergic airway disease was used to investigate which mechanism is implicated in the preventive and therapeutic actions of P17936 administered exogenously on allergen-induced bronchial inflammation and airway hyper-responsiveness , in particular focusing on the regulation of P15692 expression . RESULTS : Administration of recombinant human P17936 to OVA-inhaled mice substantially attenuated the increases in hypoxia-inducible factor ( HIF ) -α activity , P05019 production , and P15692 protein levels in the lung . In addition , the blockade of P05019 action decreased the OVA-induced P15692 expression , airway inflammation , and bronchial hyper-responsiveness . The administration of recombinant human P17936 or CBO-P11 also reduced significantly increases in inflammatory cells , airway hyper-responsiveness , levels of P05112 , P05113 , P35225 , and vascular permeability in the lung of OVA-inhaled mice . Moreover , when recombinant human P17936 was administered after the completion of OVA inhalation , these therapeutic effects of P17936 were also observed . CONCLUSIONS : These results indicate that P17936 administered exogenously may attenuate antigen-induced airway inflammation and hyper-responsiveness through the modulation of vascular leakage and P15692 expression mediated by HIF-1α/HIF-2α signaling as well as P05019 action in allergic airway disease of mice . Cloning , after cloning , knock-out mice , and physiological functions of MAO A and B . Cloning of MAO A and B has demonstrated clearly that MAO A and B are coded by different proteins with 70 % amino acid identity . With the MAO A and B cDNA clones , we showed the tissue distribution and genomic structure of MAO A and B , the latter suggesting that they are derived from the same ancestral gene . The active sites , the role of cysteine residues , the three-dimensional models and the mitochondria targeting domains of both isoenzymes have been established . The transcriptional regulation of MAO A and B has been studied . MAO A KO mice showed increased levels of serotonin ( 5-HT ) , norepinephrine ( NE ) , dopamine ( DA ) whereas MAO B KO mice showed increased phenylethylamine ( PEA ) levels only . Both MAO A and B KO mice showed increased response to stress . MAO A KO mice showed increased emotional learning and memory and aggressive behavior , but the vesicular monoamine transporter ( Q05940 ) , P08908 , 5- Q13049 and P28335 receptors were down regulated . 5- Q13049 antagonist , ketanserin and MDL100907 were able to abolish the aggression , suggesting that the aggressive behavior may be mediated by 5- Q13049 receptor . In contrast , MAO B KO mice are resistant to MPTP , a toxin which induces Parkinson 's syndromes . Studies of these mice suggest that MAO A and B have distinct biochemical and physiological functions . 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 . 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 .

MH_train_1578

MH_train_1578

MH_train_1578

interacts_with DB09068?

multiple_choice

5-HT released by mucosal stimuli initiates peristalsis by activating Q13639 /5-HT1p receptors on sensory P80511 neurons . The intestinal peristaltic reflex can be elicited by mucosal stimulation or circular muscle stretch . Muscle stretch activates extrinsic , whereas mucosal stimulation activates intrinsic calcitonin gene-related peptide ( P80511 ) -containing sensory neurons . The present study examined the role of 5-hydroxytryptamine ( 5-HT ) in sensory transmission . A three-compartment preparation of rat colon was used that enables separate measurement of sensory transmitters and modulators . Mucosal stimuli ( 2-8 brush strokes ) caused concurrent increase in 5-HT and P80511 release in proportion to the intensity of stimulation . Release of both 5-HT and P80511 occurred exclusively into the central compartment where the stimuli were applied . Exogenous 5-HT caused a concentration-dependent release of P80511 . Release of P80511 induced by exogenous 5-HT or mucosal stimulation was inhibited by selective Q13639 and 5-HT1p antagonists but was not affected by P08908 , 5-HT2 , and 5- Q9H205 antagonists . Ascending contraction and descending relaxation of circular muscle measured in the peripheral orad and caudad compartments , respectively , were also selectively inhibited by Q13639 and 5-HT1p antagonists added to the central but not peripheral compartments . In contrast , muscle stretch elicited P80511 but not 5-HT release ; the ascending contraction and descending relaxation components of the peristaltic reflex induced by muscle stretch were not affected by 5-HT antagonists . We conclude that 5-HT released by mucosal stimulation initiates the peristaltic reflex by activating Q13639 /5-HT1p receptors on sensory P80511 -containing neurons . 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 . Interactions between bradykinin ( BK ) and cell adhesion molecule ( P62158 ) expression in peptidoglycan-polysaccharide ( PG-PS ) -induced arthritis . Bradykinin ( BK ) , a vasoactive , proinflammatory nonapeptide , promotes cell adhesion molecule ( P62158 ) expression , leukocyte sequestration , inter-endothelial gap formation , and protein extravasation in postcapillary venules . These effects are mediated by bradykinin-1 ( P46663 ) and-2 ( P30411 ) receptors . We delineated some of the mechanisms by which BK could influence chronic inflammation by altering P62158 expression on leukocytes , endothelium , and synovium in joint sections of peptidoglycan-polysaccharide-injected Lewis rats . Blocking P46663 results in significantly increased joint inflammation . Immunohistochemistry of the P46663 antagonist group revealed increased leukocyte and synovial CD11b and CD54 expression and increased CD11b and P16070 endothelial expression . P30411 antagonism decreased leukocyte and synovial P16070 and CD54 and endothelial CD11b expression . Although these findings implicate P30411 involvement in the acute phase of inflammation by facilitating leukocyte activation ( CD11b ) , homing ( P16070 ) , and transmigration ( CD54 ) . Treatment with a P30411 antagonist did not affect the disease evolution in this model . In contrast , when both BK receptors are blocked , the aggravation of inflammation by P46663 blockade is neutralized and there is no difference from the disease-untreated model . Our findings suggest that P46663 and P30411 signaling show physiologic antagonism . P46663 signaling suggests involvement in down-regulation of leukocyte activation , transmigration , and homing . Further studies are needed to evaluate the B1 receptor agonist 's role in this model . Effect of various forms of the P05155 ( DB05341 ) and P08174 on complement mediated xenogeneic cell lysis . The purpose of the present study was to assess the effect of various forms of the surface-bound form of the P05155 ( DB05341 -PI ) and decay accelerating factor ( P08174 ) on xenogenic cells . cDNAs of various deletion mutants of the DB05341 -PI , such as delta-1-99 amino acid ( AA ) , delta-108-183AA loop , delta-whole loop , delta-exon5 , delta-exon6 + 7 , and delta-exon5 + 6 + 7 , and that of P08174 , the delta-short consensus repeat ( SCR ) 1- P08174 were established . While all deletion mutants of DB05341 -PI except the delta-1-99AA were expressed in the cytoplasm but not on the cell surface , the delta-1-99AA was clearly expressed on the xenogeneic cell surface . Amelioration of complement-mediated xenogeneic cell lysis by delta-1-99AA was next tested , and compared with delta-SCR1 P08174 . Both molecules blocked human complement-mediated cell lysis by approximately 57 to 90 and 93 to 98 % , respectively , in Chinese hamster ovarian tumor ( CHO ) cells and pig endothelial cells ( PECs ) . The CHO cell transfectants were incubated with 20 % normal human serum , and the amounts of C4 and P01024 deposition on the cell surface were analysed by flow cytometry . The P08174 transfectant showed a large amount of C4-deposition and much less P01024 -deposition than the controls ( approximately 85 % suppression ) , whereas the delta-1-99AA showed approximately a 40 % suppression in both C4- and P01024 -deposition . Consequently , both the delta-1-99AA DB05341 -PI and delta-SCR1 P08174 molecules are quite effective in down-regulating the xenogeneic cell lysis , but accomplished this in different manners . 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 . Switching brain serotonin with oxytocin . Serotonin ( 5-HT ) and oxytocin ( P01178 ) are two neuromodulators involved in human affect and sociality and in disorders like depression and autism . We asked whether these chemical messengers interact in the regulation of emotion-based behavior by administering P01178 or placebo to 24 healthy subjects and mapping cerebral 5-HT system by using 2'-methoxyphenyl-(N-2'-pyridinyl)-p-[(18)F]fluoro-benzamidoethylpiperazine ( [(18)F]MPPF ) , an antagonist of P08908 receptors . P01178 increased [(18)F]MPPF nondisplaceable binding potential ( BPND ) in the dorsal raphe nucleus ( DRN ) , the core area of 5-HT synthesis , and in the amygdala/hippocampal complex , insula , and orbitofrontal cortex . Importantly , the amygdala appears central in the regulation of 5-HT by P01178 : [(18)F]MPPF BPND changes in the DRN correlated with changes in right amygdala , which were in turn correlated with changes in hippocampus , insula , subgenual , and orbitofrontal cortex , a circuit implicated in the control of stress , mood , and social behaviors . P01178 administration is known to inhibit amygdala activity and results in a decrease of anxiety , whereas high amygdala activity and 5-HT dysregulation have been associated with increased anxiety . The present study reveals a previously unidentified form of interaction between these two systems in the human brain , i.e. , the role of P01178 in the inhibitory regulation of 5-HT signaling , which could lead to novel therapeutic strategies for mental disorders . 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 . Complement deposition and microglial activation in the outer retina in light-induced retinopathy : inhibition by a P08908 agonist . PURPOSE : Increasing evidence supports a role for complement in the pathogenesis of age-related macular degeneration ( AMD ) . This study evaluated retinal microglia , T-lymphocytes , and complement deposition in a light-induced retinopathy model . The effect of a serotonin ( 5-hydroxytryptamine , 5-HT(1A) ) agonist on these processes was investigated . METHODS : Rats were dark adapted for 24 hours before a 6-hour blue light exposure . Some animals were predosed subcutaneously with AL-8309A . Retinas were evaluated at different times after light exposure . Paraffin sections were stained with antibody for a microglial marker ( Iba1 ) , a T-lymphocyte marker ( CD3 ) , and complement components C1q , P01024 , factor B , factor H , and membrane attack complex ( MAC ) . RESULTS : Light exposure resulted in substantial photoreceptor and Q96AT9 loss . Robust microglia activation and migration to the outer retina occurred rapidly . Substantial T-lymphocyte recruitment did not occur . Complement alternative pathway was strongly activated , resulting in the deposition of P01024 , factor B , factor H , and MAC in the area of photic lesions . Dosing with AL-8309A prevented retinal lesions and decreased microglia activation/recruitment and complement deposition in the outer retina . CONCLUSIONS : In blue light exposed retinas , microglia were activated and migrated toward the outer retina , whereas a T-lymphocyte response was minimal . The innate immune system was markedly activated , with substantial complement deposition in the outer retina after light exposure . This complement deposition was prevented by AL-8309A . This model may be useful in the evaluation of complement inhibitors and other neuroprotectants intended for ocular use . AL-8309 is under evaluation in the clinic and may be useful in the treatment of AMD . Bradykinin facilitates noradrenaline spillover during contraction in the canine gracilis muscle . DB00368 spillover from skeletal muscle vascular areas increases during exercise but the underlying mechanisms are not well understood . Muscle contraction itself causes changes in many factors that could affect noradrenaline spillover . For instance , it has been reported that bradykinin is synthesized in skeletal muscle areas during contraction . Because the P30411 facilitates noradrenaline spillover , it may be involved in the increase associated with contraction . In this experiment , we studied the effect of bradykinin on noradrenaline spillover in the in situ canine gracilis muscle , using the specific B2 antagonist DB06196 . The drug did not modify noradrenaline spillover at rest , but did cause a significant decrease during muscle contraction , from 558 to 181 pg min(-1) . As reported previously in the literature , fractional extraction of noradrenaline decreased during muscle contraction . This effect was independent of HOE 140 treatment . In light of our results , it seems that bradykinin formation during muscle contraction may play an important part in the observed increase in noradrenaline spillover but does not affect fractional extraction . 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 . DB00074 induction in patients receiving tacrolimus-based immunosuppressive regimens . PURPOSE : The use of basiliximab induction increased significantly in recent years based on its superior efficacy and excellent safety profile demonstrated in studies with cyclosporine-based immunosuppression . However , its clinical utility in patients receiving tacrolimus-based immunosuppressive regimens is still uncertain . METHODS : We retrospectively reviewed data of 366 low immunological risk recipients of deceased donor kidney transplants . Of them , 134 received basiliximab and tacrolimus ( TAC- P01589 ) , 100 received basiliximab and delayed tacrolimus(dTAC- P01589 ) , and 132 patients received tacrolimus without basiliximab(TAC-No) . The endpoints were the incidence of acute rejection , graft function , and patient and graft survivals at 1 year . RESULTS : The incidence of acute rejection was higher in dTAC- P01589 compared to TAC-IL-2RA and TAC-No Groups ( 33 vs.14.9 vs. 14.3 % , p < 0.001 ) . Inferior creatinine clearance was observed in dTAC- P01589 Group compared to TAC- P01589 and TAC-No Groups at months 1 ( 41.6 vs. 49.9 vs. 44.8 mL/min , p = 0.004 ) , 3 ( 49.8 vs. 57.2 vs. 53.5 mL/min , p = 0.017 ) , and 6 ( 53.1 vs. 61.8 vs. 57.0 mL/min , p = 0.001 ) . Patients who received basiliximab ( TAC- P01589 and dTAC- P01589 Groups ) had lower incidence of posttransplant diabetes ( 24 vs.18 vs. 39.3 % , p = 0.009 ) . Patient and graft survivals were similar among the groups . CONCLUSIONS : In low immunological risk kidney transplant recipients receiving tacrolimus , the use of basiliximab induction was not associated with lower rejection rates and did not allow delayed tacrolimus introduction . 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 . YAP modifies cancer cell sensitivity to P00533 and survivin inhibitors and is negatively regulated by the non-receptor type protein tyrosine phosphatase 14 . The Yes-associated protein ( YAP ) is a transcriptional factor involved in tissue development and tumorigenesis . Although YAP has been recognized as a key element of the Hippo signaling pathway , the mechanisms that regulate YAP activities remain to be fully characterized . In this study , we demonstrate that the non-receptor type protein tyrosine phosphatase 14 ( Q15678 ) functions as a negative regulator of YAP . We show that YAP forms a protein complex with Q15678 through the WW domains of YAP and the PPXY motifs of Q15678 . In addition , Q15678 inhibits YAP-mediated transcriptional activities . Knockdown of YAP sensitizes cancer cells to various anti-cancer agents , such as cisplatin , the P00533 tyrosine kinase inhibitor erlotinib and the small-molecule antagonist of survivin , P28222 . YAP-targeted modalities may be used in combination with other cancer drugs to achieve maximal therapeutic effects . Effects of neuropeptides on human lung fibroblast proliferation and chemotaxis . An increase in subepithelial mesenchymal cells and associated connective tissue is a feature of bronchial asthma . We determined whether neuropeptides could modulate fibroblast activity , particularly with respect to proliferation and chemotaxis . Human lung fibroblasts were cultured with neurokinin A ( P20366 ) , DB05875 ( SP ) , vasoactive intestinal peptide ( P01282 ) , and calcitonin-gene-related peptide ( P80511 ) . After 48 h , fibroblast proliferation was measured by a colorimetric assay based on the uptake and subsequent release of methylene blue . The chemotactic response to neuropeptides was determined with the use of a modified Boyden chamber . Both P20366 and SP ( 10(-7)-10(-4) M ) stimulated human lung fibroblast proliferation in Q03591 and IMR-90 fibroblasts . P01282 and P80511 had no effect on fibroblast proliferation . P20366 alone stimulated fibroblast chemotaxis maximally at 10(-10) M . P08473 ( NEP ) activity of 0.52 and 5.2 pmol/10(6) cells was assayed in IMR-90 and Hs68 fibroblasts , respectively . DB02557 ( 5 x 10(-6)-10(-5) M ) , an NEP inhibitor , enhanced fibroblast proliferation in a dose-dependent manner . Thus neuropeptides have the potential to cause activation of mesenchymal cells , and neuropeptide release may contribute to the structural abnormalities observed in asthmatic airways . Deactivation of the lipopolysaccharide antagonist eritoran ( E5564 ) by high-density lipoprotein-associated apolipoproteins . Lipid A , the active moiety of LPS , exerts its effects through interaction with O00206 , triggering a signalling cascade that results in the release of pro-inflammatory cytokines . DB04933 is a lipid A analogue that competes with LPS for binding to O00206 ; however , after intravenous administration , it undergoes a time-dependent deactivation as a consequence of binding to high-density lipoproteins ( HDLs ) . The site of eritoran association with HDL remains unknown . Therefore the aim of this study was to determine if HDL-associated apolipoproteins A1 , A2 , serum amyloid A ( P0DJI8 ) and C1 , inhibit the ability of eritoran to block LPS-induced P01375 -α release from whole blood . DB04933 activity after LPS stimulation in human whole blood was assessed in the presence of reconstituted HDL ( rHDL ) containing different apos . In rHDL , the major apolipoproteins in both the healthy and septic state , A1 and P0DJI8 , caused a significant reduction in eritoran antagonistic activity and had a greater effect than minor apolipoproteins A2 and C1 . Apolipoproteins associated with HDL are likely to facilitate eritoran deactivation . Apolipoproteins A1 and P0DJI8 should be of particular focus as they are the major apos found on HDL in both the healthy and septic state . Further evaluation of the physical association between apolipoproteins and eritoran should be explored . 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 . Twelve loci from HSA10 , HSA11 and HSA20 were comparatively Q5TCZ1 -mapped on river buffalo and sheep chromosomes . Ten type I loci from HSA10 ( P01589 and P08670 ) , HSA11 ( P68871 and P01225 ) and HSA20 ( P07204 , AVP/ P01178 , GNAS1 , P08631 and P11387 ) and two domestic cattle type II loci ( CSSM30 and BL42 ) were Q5TCZ1 mapped to R-banded river buffalo ( BBU ) and sheep ( OAR ) chromosomes . P01589 ( HSA10 ) maps on BBU14q13 and OAR13q13 , P08670 ( HSA10 ) maps on BBU14q15 and OAR13q15 , P68871 ( HSA11 ) maps on BBU16q25 and OAR15q23 , P01225 ( HSA11 ) maps on BBU16q28 and OAR15q26 , P07204 ( HSA20 ) maps on BBU14q15 and OAR13q15 while AVP/ P01178 , GNAS1 , P08631 , and P11387 ( HSA20 ) as well as CSSM30 and BL42 map on the same large band of BBU14q22 and OAR13q22 . All loci were mapped on the same homologous chromosomes and chromosome bands of the two species , and these results agree with those earlier reported in cattle homologous chromosomes 15 and 13 , respectively , confirming the high degree of both banding and physical map similarities among the bovid species . Indirect comparisons between physical maps achieved on bovid chromosomes and those reported on HSA10 , HSA11 and HSA20 were performed . 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 . Development and characterization of 89Zr-labeled panitumumab for immuno-positron emission tomographic imaging of the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) is overexpressed in the majority of malignancies and has been associated with poor outcomes . DB01269 , an anti- P00533 monoclonal antibody that binds to the extracellular binding domain of P00533 , is increasingly used with radiotherapy and chemotherapy but has associated toxicities . The purpose of this study was to develop and characterize a novel targeted imaging agent for the P00533 using radiolabeled panitumumab . Flow cytometry studies were performed to evaluate P00533 expression in several cell lines . DB00746 -Bz-NCS ( DB00746 ) was conjugated to panitumumab and labeled with (89)Zr . Cell uptake studies were performed in four cell lines . For biodistribution studies and micro-positron emission tomography/computed tomography ( PET/CT ) , mouse xenograft models were generated using the same cell lines . PET was performed , and tumors and select organs were harvested for biodistribution studies . DB01269 was radiolabeled with (89)Zr with high radiochemical purity and specific activity and was found to be stable in serum . Cell binding studies demonstrated that radiotracer uptake in cells correlated with the degree of P00533 expression . MicroPET/CT imaging studies demonstrated a high intensity of (89)Zr-panitumumab in A431 and HCT 116 tumors in comparison with the P00533 -negative tumors . Biodistribution studies confirmed the results from the imaging studies . (89)Zr-panitumumab imaging of P00533 -positive tumors demonstrated levels of radiotracer uptake associated with P00533 expression . 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 . Intra- and interindividual epigenetic variation in human germ cells . Epigenetics represents a secondary inheritance system that has been poorly investigated in human biology . The objective of this study was to perform a comprehensive analysis of DNA methylation variation between and within the germlines of normal males . First , methylated cytosines were mapped using bisulphite modification-based sequencing in the promoter regions of the following disease genes : presenilins ( P49768 and P49810 ) , breast cancer ( P38398 and P51587 ) , myotonic dystrophy ( DM1 ) , and Huntington disease ( HD ) . Major epigenetic variation was detected within samples , since the majority of sperm cells of the same individual exhibited unique DNA methylation profiles . In the interindividual analysis , 41 of 61 pairwise comparisons revealed distinct DNA methylation profiles ( P=.036 to 6.8 x 10(-14) ) . Second , a microarray-based epigenetic profiling of the same sperm samples was performed using a 12,198-feature CpG island microarray . The microarray analysis has identified numerous DNA methylation-variable positions in the germ cell genome . The largest degree of variation was detected within the promoter CpG islands and pericentromeric satellites among the single-copy DNA fragments and repetitive elements , respectively . A number of genes , such as O75530 , P26232 , P62158 , P55290 , and Q93045 , exhibited age-related DNA methylation changes . Finally , allele-specific methylation patterns in P55290 were detected . This study provides evidence for significant epigenetic variability in human germ cells , which warrants further research to determine whether such epigenetic patterns can be efficiently transmitted across generations and what impact inherited epigenetic individuality may have on phenotypic outcomes in health and disease .

MH_train_1579

MH_train_1579

MH_train_1579

interacts_with DB00007?

multiple_choice

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 . DB08916 and lung cancer . P00533 tyrosine kinase inhibitors ( P00533 -TKI ) have an established role in the treatment of non-small-cell lung cancer ( NSCLC ) . First-generation reversible DB00171 -competitive P00533 -TKIs are approved for the initial treatment of patients with P00533 mutation-positive advanced NSCLC . DB08916 is an irreversible second-generation P00533 -TKI with potent preclinical activity against P00533 ( wild type and mutant ) , P04626 , Q15303 and P00533 -mutant NSCLC with acquired resistance to reversible P00533 -TKI . LUX-Lung 3 trial demonstrated superiority of afatinib to cisplatin and pemetrexed in the frontline treatment of treatment-naïve patients with advanced adenocarcinoma of the lung and P00533 mutation . Based on these results , afatinib was recently approved for the first-line treatment of NSCLC patients with P00533 mutation . This article summarizes current status of preclinical and clinical development of afatinib in NSCLC . Phosphorylation of Grainy head by P29323 is essential for wound-dependent regeneration but not for development of an epidermal barrier . Grainy head ( P01148 ) is a key transcription factor responsible for epidermal barrier formation and repair , whose function is highly conserved across diverse animal species . However , it is not known how P01148 function is reactivated to repair differentiated epidermal barriers after wounding . Here , we show that P01148 is directly regulated by extracellular signal-regulated kinase ( P29323 ) phosphorylation , which is required for wound-dependent expression of P01148 target genes in epidermal cells . DB00133 91 is the principal residue in P01148 that is phosphorylated by P29323 . Although mutations of the P29323 phosphorylation sites in P01148 do not impair its DNA binding function , the P29323 sites in P01148 are required to activate Dopa decarboxylase ( Ddc ) and misshapen ( msn ) epidermal wound enhancers as well as functional regeneration of an epidermal barrier upon wounding . This result indicates that the phosphorylation sites are essential for damaged epidermal barrier repair . However , P01148 with mutant P29323 phosphorylation sites can still promote barrier formation during embryonic epidermal development , suggesting that P29323 sites are dispensable for the P01148 function in establishing epidermal barrier integrity . These results provide mechanistic insight into how tissue repair can be initiated by posttranslational modification of a key transcription factor that normally mediates the developmental generation of that tissue . Targeted chemotherapy for triple-negative breast cancers via P01148 receptor . Triple-negative breast cancer does not express estrogen and progesterone receptors and there is no overexpression/amplification of the P04626 -neu gene . Therefore , this subtype of breast cancer lacks the benefits of specific therapies which target these receptors . About 60 % of all human breast cancers express receptors for luteinizing hormone releasing hormone ( P01148 , DB00644 ) , which might be used as a target . The P01148 receptor can be used for targeted chemotherapy with cytotoxic luteinizing hormone releasing hormone agonists such as AEZS-108 ( AN-152 ) , in which doxorubicin is linked to [D-Lys6] P01148 . In the present study we have analyzed by in vitro and in vivo experiments whether the cytotoxic P01148 agonist AEZS-108 ( AN-152 ) induces apoptosis in triple-negative human breast cancer cells that express P01148 receptors . P01148 receptor expression in tumor biopsy specimens of triple-negative breast cancers was tested using immunohistochemistry . Cell proliferation was analyzed using alamar blue proliferation assay . Induction of apoptosis was quantified by measurement of loss of mitochondrial membrane potential . In vivo experiments were performed using nude mice bearing xenografted human breast tumors.Thirty-one of 42 triple-negative breast cancers ( 73.8 % ) expressed P01148 receptors . We could show that treatment of triple-negative but P01148 -positive MDA-MB-231 , HCC1806 and HCC1937 human breast cancer cells with AEZS-108 ( AN-152 ) resulted in apoptotic cell death in vitro via activation of caspase-3 . The antitumor effects were confirmed in nude mice . AEZS-108 ( AN-152 ) inhibited the growth of xenotransplants of triple-negative human breast cancers in nude mice completely , without any apparent side effects . The cytotoxic P01148 agonist AEZS-108 ( AN-152 ) seems to be a suitable drug for an efficacious therapy for triple-negative breast cancers with little toxicity . Enhanced vasoactive intestinal peptide-induced prolactin secretion from anterior pituitary cells of incubating turkeys ( Meleagris gallopavo ) . During incubation , female turkeys exhibit elevated circulating prolactin ( PRL ) which may be the result of enhanced pituitary responsiveness to vasoactive intestinal peptide ( P01282 ) . This hypothesis was tested by comparison of spontaneous and porcine P01282 -induced PRL secretion from anterior pituitary cells of hens in various reproductive conditions . The effect of P01282 and luteinizing hormone releasing hormone ( P01148 ) , alone and in combination , on luteinizing hormone ( LH ) secretion was also examined . Incubation with pVIP ( 10(-10) to 10(-6) M ) significantly stimulated PRL secretion at all incubation times tested ( 1-5 hr ) . This increase was greatest in cells from incubating hens , with those from laying , photorefractory , and quiescent ( nonphotostimulated ) hens secreting successively less PRL . These responses were obtained when spontaneous PRL secretions were compared . P01282 induced approximately a similar 1.5-fold increase in LH secretion , in all reproductive groups . Also , P01282 enhanced P01148 -induced LH secretion ( 1.2- to 1.6-fold ; P less than 0.0001 ) . It is concluded that PRL secretion in vitro by pituitary cells from turkey hens in various reproductive stages reflects the circulating levels of PRL at these stages . Serotonin-immunoreactive axon terminals innervate pyramidal cells and interneurons in the rat basolateral amygdala . The basolateral nuclear complex of the amygdala ( O43927 ) receives a dense serotonergic innervation that appears to play a critical role in the regulation of mood and anxiety . However , little is known about how serotonergic inputs interface with different neuronal subpopulations in this region . To address this question , dual-labeling immunohistochemical techniques were used at the light and electron microscopic levels to examine inputs from serotonin-immunoreactive ( 5-HT+ ) terminals to different neuronal subpopulations in the rat O43927 . Pyramidal cells were labeled by using antibodies to calcium/calmodulin-dependent protein kinase II , whereas different interneuronal subpopulations were labeled by using antibodies to a variety of interneuronal markers including parvalbumin ( PV ) , vasoactive intestinal peptide ( P01282 ) , calretinin , calbindin , cholecystokinin , and somatostatin . The O43927 exhibited a dense innervation by thin 5-HT+ axons . Electron microscopic examination of the anterior basolateral nucleus ( BLa ) revealed that 5-HT+ axon terminals contained clusters of small synaptic vesicles and a smaller number of larger dense-core vesicles . Serial section reconstruction of 5-HT+ terminals demonstrated that 76 % of these terminals formed synaptic junctions . The great majority of these synapses were symmetrical . The main targets of 5-HT+ terminals were spines and distal dendrites of pyramidal cells . However , in light microscopic preparations it was common to observe apparent contacts between 5-HT+ terminals and all subpopulations of O43927 interneurons . Electron microscopic analysis of the BLa in sections dual-labeled for 5-HT/PV and 5-HT/ P01282 revealed that many of these contacts were synapses . These findings suggest that serotonergic axon terminals differentially innervate several neuronal subpopulations in the O43927 . Andrographis paniculata extract ( DB05767 ) for active ulcerative colitis . OBJECTIVES : Andrographis paniculata has in vitro inhibitory activity against P01375 -α , IL-1β and NF-κB . A pilot study of A. paniculata extract ( DB05767 ) suggested similar efficacy to mesalamine for ulcerative colitis . METHODS : A randomized , double-blind , placebo-controlled trial evaluated the efficacy of A. paniculata extract ( DB05767 ) in 224 adults with mild-to-moderate ulcerative colitis . Patients were randomized to A. paniculata extract ( DB05767 ) 1,200 mg or 1,800 mg daily or placebo for 8 weeks . RESULTS : In total , 45 and 60 % of patients receiving A. paniculata 1,200 mg and 1,800 mg daily , respectively , were in clinical response at week 8 , compared with 40 % of those who received placebo ( P=0.5924 for 1,200 mg vs. placebo and P=0.0183 for 1,800 mg vs. placebo ) . In all , 34 and 38 % of patients receiving A. paniculata 1,200 mg and 1,800 mg daily , respectively , were in clinical remission at week 8 , compared with 25 % of those who received placebo ( P=0.2582 for 1,200 mg vs. placebo and P=0.1011 for 1,800 mg vs. placebo ) . Adverse events developed in 60 and 53 % of patients in the A. paniculata 1,200 mg and 1,800 mg daily groups , respectively , and 60 % in the placebo group . CONCLUSIONS : Patients with mildly to moderately active ulcerative colitis treated with A. paniculata extract ( DB05767 ) at a dose of 1,800 mg daily were more likely to achieve clinical response than those receiving placebo . 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 . Estrogen-astrocyte-luteinizing hormone-releasing hormone signaling : a role for transforming growth factor-beta(1) . The purpose of this study was to identify factors from astrocytes that can regulate P01148 neurosecretion . Exposure of P01148 -secreting ( GT1-7 ) cells to conditioned media ( CM ) from P13671 glial cells and hypothalamic astrocytes ( HA ) stimulated P01148 release . Assays of P13671 and HA CM revealed that transforming growth factor-beta(1) ( TGF-beta(1) ) and 3alpha-hydroxy-5alpha-pregnane-20-one ( 3alpha , 5alpha-THP ) , both known P01148 secretagogues , were present in CM and their levels increased in parallel to the P01148 -releasing activity of CM . In contrast , TGF-alpha was undetectable in P13671 or HA CM . Ultrafiltration to remove peptides with molecular weights > 10 kDa virtually abolished the P01148 -releasing ability of the HA CM . Furthermore , immunoneutralization with a panspecific DB00116 -beta antibody dose-dependently attenuated the P01148 -releasing activity of the CM . Rat hypothalamus and GT1-7 cells were demonstrated to express TGF-beta receptors as well as furin , an enzyme that converts latent TGF-beta(1) to active TGF-beta(1) . P03372 -alpha and Q92731 mRNA and protein were also demonstrated in HAs by reverse transcription-polymerase chain reaction and double immunofluorescence , and treatment with 17beta-estradiol ( 17beta-E(2) ) increased both active and latent TGF-beta(1) levels in HA CM . The effect of 17beta-E(2) was completely blocked by the ER antagonist ICI8280 . As a whole , these studies provide evidence of a previously undescribed 17beta-E(2)-TGF-beta(1)- P01148 signaling pathway . 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 . Toll-like receptor stimulation differentially regulates vasoactive intestinal peptide type 2 receptor in macrophages . Vasoactive intestinal peptide ( P01282 ) was originally isolated as a vasodilator intestinal peptide , then as a neuropeptide . In the immune system , P01282 is described as an endogenous macrophage-deactivating factor . P01282 exerts its immunological actions in a paracrine and/or autocrine manner , through specific receptors . However , very little is known about the molecular regulation of P01282 type 2 receptor ( VPAC(2) ) in the immune system . We now report that different toll-like receptor ( TLR ) ligands selectively regulate the VPAC(2) receptor gene and show a gene repression system controlled by key protein kinase signalling cascades in macrophages . VPAC(2) gene expression is regulated by gram-positive ( O60603 ligands ) and gram-negative bacteria wall constituents ( O00206 ligands ) . Moreover , VPAC(2) is tightly regulated : O60603 - or O60603 /6- but not O60603 /1-mediated mechanisms are responsible for the induction of VPAC(2) . TLR stimulation by viral or bacterial nucleic acids did not modify the VPAC(2) mRNA levels . Remarkably , imiquimod -- a synthetic Q9NYK1 ligand -- led to a potent up-regulation of VPAC(2) gene expression . O60602 stimulation by flagellin present in gram-positive and gram-negative bacteria did not affect VPAC(2) mRNA . The p38 mitogen-activated protein kinase ( MAPK ) activity accounted for the O00206 -mediated induction of VPAC(2) gene expression . Surprisingly , our data strongly suggest for the first time a tightly repressed control of VPAC(2) mRNA induction by elements downstream of MAPK kinase 1/2 , PI3K/Akt , and particularly Jun-NH(2)-terminal kinase signalling pathways . 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 . DB00644 antagonist cetrorelix down-regulates proliferating cell nuclear antigen and epidermal growth factor expression and up-regulates apoptosis in association with enhanced poly ( adenosine 5'-diphosphate-ribose ) polymerase expression in cultured human leiomyoma cells . The objective of this study was to elucidate the effects of DB00644 antagonist DB00050 on proliferation and apoptosis in human leiomyoma cells cultured in vitro . Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10 % fetal bovine serum for 120 h and then stepped down to serum-free conditions in the presence or absence of graded concentrations of DB00050 ( 10(-5) to 10(-8) mol/liter ) for 6 d . Cultured leiomyoma cells were used for semiquantitative RT-PCR , immunocytochemistry , Western blot analysis , and terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling assay . RT-PCR analysis revealed the presence of mRNAs encoding for P30968 and epidermal growth factor ( P01133 ) in cultured leiomyoma cells . The number of viable cultured leiomyoma cells was significantly ( P < 0.01 ) decreased by treatment with DB00050 compared with untreated control cultures . Immunocytochemical examination demonstrated that treatment with DB00050 attenuated the expression of proliferating cell nuclear antigen ( P12004 ) and P01133 in cultured leiomyoma cells . Western blot analysis revealed that treatment with 10(-5) mol/liter DB00050 significantly ( P < 0.01 ) decreased P12004 expression . In addition , treatment with 10(-5) mol/liter DB00050 remarkably increased the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling-positive rate and poly(ADP-ribose) polymerase expression at 24 h of treatment compared with untreated control cultures ( P < 0.01 ) . Furthermore , treatment with 10(-5) mol/liter DB00050 decreased immunoreactive P01133 protein and P01133 mRNA expression in cultured leiomyoma cells at 4 d of treatment . DB00644 antagonist DB00050 may directly inhibit leiomyoma cell growth by down-regulating proliferation in association with a decrease in P01133 mRNA expression and by up-regulating apoptosis in those cells . Inhibitory effects of a luteinizing hormone-releasing hormone agonist on basal and epidermal growth factor-induced cell proliferation and metastasis-associated properties in human epidermoid carcinoma A431 cells . The purpose of this study was to investigate the effects of a potent P01148 agonist , [D- DB00150 (6)] P01148 on the basal and P01133 -induced cell proliferation and the metastasis-associated properties in A431 human epidermoid carcinoma . [D- DB00150 (6)] P01148 time-dependently inhibited the basal and P01133 -stimulated growth of A431 cancer cells . It is assumed that phosphorylation/dephosphorylation of cellular proteins is highly related to cell growth . This study demonstrates that [D- DB00150 (6)] P01148 decreased the basal and P01133 -induced total cellular kinase activity , particularly the tyrosine phosphorylation of several cellular proteins including the P00533 . In contrast , [D- DB00150 (6)] P01148 did not cause detectable changes in basal and P01133 -stimulated serine/threonine phosphorylation of A431 cellular proteins . The inhibitory effect of [D- DB00150 (6)] P01148 on A431 cell proliferation was associated with apoptosis as evidenced by the cell morphology and DNA integrity ( ladder pattern ) , the expression of interleukin 1beta-converting enzyme ( ICE ) and activation of caspase . Furthermore , P01133 could rescue the remaining attached A431 cells following [D- DB00150 (6)] P01148 treatment for 48 hr , which suggests that limited exposure to [D- DB00150 (6)] P01148 did not channel all cells to irreversible apoptotic process . We also determined the effects of [D- DB00150 (6)] P01148 on metastasis-associated properties in A431 cells . [D- DB00150 (6)] P01148 reduced both basal and P01133 -stimulated secretion of P14780 and P08253 . In addition , [D- DB00150 (6)] P01148 suppressed the basal and P01133 -induced invasive activity of A431 cells based on an in vitro invasion assay . In conclusion , this study indicates that [D- DB00150 (6)] P01148 may act partly through activating tyrosine phosphatase activity to inhibit cell proliferation and the metastasis-associated properties of A431 cancer cells . Our work suggests that [D- DB00150 (6)] P01148 may be therapeutically useful in limiting the tumor growth and metastasis of some neoplasms . DB00674 ameliorates the impairment of recognition memory in mice repeatedly treated with methamphetamine : involvement of allosteric potentiation of nicotinic acetylcholine receptors and dopaminergic- P27361 /2 systems . DB00674 , a drug used to treat Alzheimer 's disease , inhibits acetylcholinesterase ( P22303 ) and allosterically modulates nicotinic acetylcholine receptors ( nAChRs ) resulting in stimulation of catecholamine neurotransmission . In this study , we investigated whether galantamine exerts cognitive-improving effects through the allosteric modulation of nAChRs in an animal model of methamphetamine ( Meth ) psychosis . The mice treated with Meth ( 1 mg/kg.d ) for 7 d showed memory impairment in a novel object recognition test . DB00674 ( 3 mg/kg ) ameliorated the memory impairment , and it increased the extracellular dopamine release in the prefrontal cortex ( P27918 ) of Meth-treated mice . Donepezil , an P22303 inhibitor ( 1 mg/kg ) increased the extracellular ACh release in the P27918 , whereas it had no effect on the memory impairment in Meth-treated mice . The nAChR antagonist , mecamylamine , and dopamine D1 receptor antagonist , P35240 23390 , blocked the ameliorating effect of galantamine on Meth-induced memory impairment , whereas the muscarinic AChR antagonist , scopolamine , had no effect . The effects of galantamine on extracellular dopamine release were also antagonized by mecamylamine . DB00674 attenuated the defect of the novelty-induced activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . The ameliorating effect of galantamine on recognition memory in Meth-treated mice was negated by microinjection of an P29323 inhibitor , PD98059 , into the P27918 . These results suggest that the ameliorating effect of galantamine on Meth-induced memory impairment is associated with indirect activation of dopamine D1 receptor- P27361 /2 following augmentation with dopaminergic neurotransmission in the P27918 through the allosteric activation of nAChRs . DB00674 could be a useful therapeutic agent for treating cognitive deficits in schizophrenia/Meth psychosis , as well as Alzheimer 's disease . 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 . The use of a cyclooxygenase-2 inhibitor ( DB06802 ) in an ocular and metastatic animal model of uveal melanoma . The expression of cyclooxygenase-2 ( P35354 ) has been reported as an indicator of poor prognosis in a wide variety of human tumors , including colon , breast and uveal melanoma ( UM ) . P35354 inhibitors have shown promise in controlling the malignancy of several types of tumors . Previous studies have demonstrated the efficacy of a P35354 inhibitor on the proliferation rates of human UM cells . The goal of this experiment was to investigate the efficiency of DB06802 , a topically administered P35354 inhibitor , in a rabbit model of UM . The animals were divided into two groups of 14 animals for the duration of the 12-week experiment . One animal per group was killed each week to evaluate disease progression and for histopathological studies . The experimental group received drops containing 0.3 % DB06802 solution . Intraocular tumor growth was evaluated weekly by fundoscopic examination and each animal was weighed prior to examination . Blood samples were taken weekly from all rabbits to detect circulating malignant cells ( CMCs ) throughout the experiment . After the second week of inoculation , the experimental group weighed significantly more than the control group . The control group developed more intraocular tumors and presented with metastases and higher detectable levels of CMCs before the treated group . These results indicate that the topical administration of a P35354 inhibitor delayed the progression of this malignancy in our animal model . A clinical trail using an anti- P35354 inhibitor for patients with UM should be considered . 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 . 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 .

MH_train_1580

MH_train_1580

MH_train_1580

interacts_with DB00682?

multiple_choice

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 . 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 . Selection of mouse strains showing high and low incidences of alloxan-induced diabetes . To produce an experimental model of diabetes in animals , ICR mice were inbred until the 20th generation by two-way selection toward the high- and low-incidences of alloxan-induced diabetes . Changes in successive generations in the incidence of such diabetes , in blood glucose levels , growth patterns and reproductive performance were studied . The incidence of alloxan-induced diabetes was 41.1 % in the basal population ; in the high-incidence strain , it was 98.7 % in F13 , ranging between 90 and 99 % in later generations ; and in the low-incidence strain , it reached 0 % in P08709 , remaining near that level in later generations . The heritability of the incidence of alloxan-induced diabetes determined at the beginning of selection was 50-60 % . The blood glucose level was 251 +/- 19 mg/dl in the basal population ; in the high-incidence strain , it was 423 +/- 11 mg/dl in F13 , ranging thereafter between 340 and 455 mg/dl ; and in the low-incidence strain , it was 128 +/- 4 mg/dl in P08709 , then varying from 120 to 140 mg/dl in following generations . The heritability of the blood glucose level determined at the beginning of selection was 40-60 % . No marked decrease in growth or reproductive performance accompanied successive selections . Successive generations of the high-incidence mice , however , tended to become heavier than the low-incidence animals . The high- and low-incidence strains , established in the 20th generation , were named the P35858 ( alloxan-induced diabetes-susceptible ) and ALR ( alloxan-induced diabetes-resistant ) strains , respectively . Common genetic variants contribute to primary hypertriglyceridemia without differences between familial combined hyperlipidemia and isolated hypertriglyceridemia . BACKGROUND : The majority of hypertriglyceridemias are diagnosed as familial combined hyperlipidemia ( FCHL ) and primary isolated hypertriglyceridemias . The contribution of common genetic variants in primary hypertriglyceridemias and the genetic difference between FCHL and isolated hypertriglyceridemias have not been thoroughly examined . METHODS AND RESULTS : This study involved 580 patients with hypertriglyceridemias and 403 controls . Of the 37 single nucleotide polymorphisms examined , 12 located in 10 genes showed allelic and genotype frequency differences between hypertriglyceridemias and controls . The minor alleles of P02649 , Q6Q788 , GALNTN2 , and Q14397 variants were positively correlated with plasma triglycerides , whereas minor alleles of Q86V24 , Q9Y5C1 , P06858 , and Q96RU8 polymorphisms were inversely associated . Body mass index , glucose , sex , rs328 and rs7007797 in P06858 , rs662799 and rs3135506 in Q6Q788 , and rs1260326 in Q14397 explained 36 % of the variability in plasma triglycerides , 7.3 % of which was attributable to the genetic variables . P06858 , Q14397 , and Q6Q788 polymorphisms fit dominant , recessive , and additive inheritance models , respectively . Variants more frequently identified in isolated hypertriglyceridemias were rs7412 in P02649 and rs1800795 in P05231 ; rs2808607 in P22680 and rs3812316 and rs17145738 in Q9NP71 were more frequent in FCHL . The other 32 single nucleotide polymorphisms presented similar frequencies between isolated hypertriglyceridemias and FCHL . CONCLUSIONS : Common genetic variants found in P06858 , Q6Q788 , and Q14397 are associated with triglycerides levels in patients with primary hypertriglyceridemias . FCHL and isolated hypertriglyceridemias are probably trace to an accumulation of genetic variants predisposing to familial and sporadic hypertriglyceridemias or to hypertriglyceridemias and hypercholesterolemia in case of FCHL . Generation and characterization of a human monoclonal IgM antibody that recognizes a conserved epitope shared by lipopolysaccharides of different gram-negative bacteria . A hybridoma cell line secreting a human monoclonal antibody ( humab ) directed to an epitope in the lipid A region of lipopolysaccharides of Gram-negative bacteria was isolated . Peripheral blood lymphocytes ( PBL ) obtained from a healthy volunteer were immortalized by Epstein-Barr virus ( EBV ) transformation . Lymphoblastoid cell lines ( LCL ) secreting antibodies to the lipopolysaccharides of Gram-negative bacteria were determined by an enzyme-linked immunosorbent assay ( ELISA ) and subsequently fused with the human-mouse heteromyeloma cell line CB- P08709 by polyethylenglycol ( PEG ) -mediated fusion . A hybridoma line producing a humab ( LPD5H4 ) , of the IgM/lambda isotype , which strongly reacted with the lipid A portion of Salmonella and E. coli spp. in ELISA , was established . The antibody was purified by hydrophobic interaction chromatography and gel filtration . Immunoblotting experiments showed a strong reactivity of the humab LPD5H4 with the lower molecular species of different rough and smooth lipopolysaccharide ( LPS ) types of the bacteria species Salmonella , E. coli , Klebsiella , and Neisseria meningitidis , whereas those of Pseudomonas spp. were negative . Binding of humab LPD5H4 to solid phase bound lipid A and different rough mutants of LPS could be inhibited by the corresponding antigens in solution . Competition assays with a murine monoclonal antibody to lipid A and with polymyxin B indicate that humab LPD5H4 recognizes its epitope in this extremely conserved part of the LPS molecule . In vitro tests demonstrated that the MAb is able to partially inhibit the LPS-induced release of P01375 using isolated peripheral blood mononuclear cells ( PBMC ) . P41134 enhances docetaxel cytotoxicity in prostate cancer cells through inhibition of P38936 . To identify potential mechanisms underlying prostate cancer chemotherapy response and resistance , we compared the gene expression profiles in high-risk human prostate cancer specimens before and after neoadjuvant chemotherapy and radical prostatectomy . Among the molecular signatures associated with chemotherapy , transcripts encoding inhibitor of DNA binding 1 ( P41134 ) were significantly upregulated . The patient biochemical relapse status was monitored in a long-term follow-up . Patients with P41134 upregulation were found to be associated with longer relapse-free survival than patients without P41134 increase . This in vivo clinical association was mechanistically investigated . The chemotherapy-induced P41134 upregulation was recapitulated in the prostate cancer cell line LNCaP . DB01248 dose-dependently induced P41134 transcription , which was mediated by P41134 promoter E-box chromatin modification and c-Myc binding . Stable P41134 overexpression in LNCaP increased cell proliferation , promoted G(1) cell cycle progression , and enhanced docetaxel-induced cytotoxicity . These changes were accompanied by a decrease in cellular mitochondria content , an increase in P10415 phosphorylation at serine 70 , caspase-3 activation , and poly(ADP-ribose) polymerase cleavage . In contrast , P41134 siRNA in the LNCaP and C42B cell lines reduced cell proliferation and decreased docetaxel-induced cytotoxicity by inhibiting cell death . P41134 -mediated chemosensitivity enhancement was in part due to P41134 suppression of P38936 . Overexpression of P38936 in LNCaP- P41134 -overexpressing cells restored the P38936 level and reversed P41134 -enhanced chemosensitivity . These molecular data provide a mechanistic rationale for the observed in vivo clinical association between P41134 upregulation and relapse-free survival . Taken together , it shows that P41134 expression has a novel therapeutic role in prostate cancer chemotherapy and prognosis . 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 glutamate/neutral amino acid transporter family Q99705 : molecular , physiological and pharmacological aspects . The solute carrier family 1 ( Q99705 ) includes five high-affinity glutamate transporters , P43005 , GLT-1 , P43003 , P48664 and O00341 ( P43005 , P43004 , P43003 , P48664 , and O00341 , respectively ) as well as the two neutral amino acid transporters , P43007 and Q15758 ( P43007 and ALC1A5 , respectively ) . Although each of these transporters have similar predicted structures , they exhibit distinct functional properties which are variations of a common transport mechanism . The high-affinity glutamate transporters mediate transport of l- DB00142 , l- DB00128 and d- DB00128 , accompanied by the cotransport of 3 Na(+) and 1 H(+) , and the countertransport of 1 K(+) , whereas ASC transporters mediate Na(+)-dependent exchange of small neutral amino acids such as Ala , DB00133 , DB00151 and DB00156 . The unique coupling of the glutamate transporters allows uphill transport of glutamate into cells against a concentration gradient . This feature plays a crucial role in protecting neurons against glutamate excitotoxicity in the central nervous system . During pathological conditions , such as brain ischemia ( e.g. after a stroke ) , however , glutamate exit can occur due to " reversed glutamate transport " , which is caused by a reversal of the electrochemical gradients of the coupling ions . Selective inhibition of the neuronal glutamate transporter P43005 ( P43005 ) may be of therapeutic interest to block glutamate release from neurons during ischemia . On the other hand , upregulation of the glial glutamate transporter P43004 ( P43004 ) may help protect motor neurons in patients with amyotrophic lateral sclerosis ( P35858 ) , since loss of function of P43004 has been associated with the pathogenesis of certain forms of P35858 . Q07869 gene variants influence progression of coronary atherosclerosis and risk of coronary artery disease . BACKGROUND : Q07869 ( PPARalpha ) regulates the expression of genes involved in lipid metabolism and inflammation , making it a candidate gene for atherosclerosis and ischemic heart disease ( IHD ) . METHODS AND RESULTS : We investigated the association between the leucine 162 to valine ( L162V ) polymorphism and a G to C transversion in intron 7 of the PPARalpha gene and progression of atherosclerosis in the DB01241 Coronary Angiography Trial ( LOCAT ) , a trial examining the effect of gemfibrozil treatment on progression of atherosclerosis after bypass surgery and on risk of IHD in the second Northwick Park Heart Study ( Q9NP85 ) , a prospective study of healthy middle-aged men in the United Kingdom . There was no association with plasma lipid concentrations in either study . Both polymorphisms influenced progression of atherosclerosis and risk of IHD . V162 allele carriers had less progression of diffuse atherosclerosis than did L162 allele homozygotes with a similar trend for focal atherosclerosis . Intron 7 C allele carriers had greater progression of atherosclerosis than did G allele homozygotes . The V162 allele attenuated the proatherosclerotic effect of the intron 7 C allele . Homozygotes for the intron 7 C allele had increased risk of IHD , an effect modulated by the L162V polymorphism CONCLUSIONS : The PPARalpha gene affects progression of atherosclerosis and risk of IHD . Absence of association with plasma lipid concentrations suggests that PPARalpha affects atherosclerotic progression directly in the vessel wall . Brominated cyclodipeptides from the marine sponge Geodia barretti as selective 5-HT ligands . The brominated cyclodipeptides barettin ( cyclo[(6-bromo-8-entryptophan)arginine] ) and 8,9-dihydrobarettin ( cyclo[(6-bromotryptophan)arginine] ) isolated from the marine sponge Geodia barretti have previously been shown to inhibit settlement of barnacle larvae in a dose-dependent manner in concentrations ranging from 0.5 to 25 microM . To further establish the molecular target and mode of action of these compounds , we investigated their affinity to human serotonin receptors . The tryptophan residue in the barettins resembles that of endogenous serotonin [ 5-hydroxytryptamine ] . A selection of human serotonin receptors , including representatives from all subfamilies ( 1-7 ) , were transfected into P29320 -293 cells . Barettin selectively interacted with the serotonin receptors 5- Q13049 , P28335 , and Q13639 at concentrations close to that of endogenous serotonin , with the corresponding Ki values being 1.93 , 0.34 , and 1.91 microM , respectively . 8,9-Dihydrobarettin interacted exclusively with the P28335 receptor with a Ki value of 4.63 microM ; it failed to show affinity to 5- Q13049 and Q13639 , indicating that the double bond between the tryptophan and arginine residue plays an important role in the interaction with the receptor proteins . Involvement of alpha1beta1 integrin in insulin-like growth factor-1-mediated protection of PC12 neuronal processes from tumor necrosis factor-alpha-induced injury . P08069 ( IGF-1R ) supports neuronal survival against a wide variety of insults . This includes tumor necrosis factor-alpha ( TNFalpha ) -mediated neuronal damage , which represents one of the factors suspected to play a role in HIV-associated dementia ( HAD ) . PC12 neurons engineered to express human IGF-1R ( PC12/IGF-1R ) maintain neuronal processes on collagen IV for several weeks . However , prolonged treatment with TNFalpha caused degeneration of neuronal processes , with no apparent signs of apoptosis . In this process , TNFalpha did not affect DB01277 -mediated phosphorylation of P35568 , Q9Y4H2 , Akt , or Erks . In addition , PC12/IGF-1R cells were found to express predominantly alpha1beta1 integrin , which has high affinity to collagen IV . The treatment of PC12/IGF-1R neurons with a specific alpha1beta1 integrin inhibitor , obtustatin , also caused loss of neuronal processes , accompanied by a quick cell detachment and extensive apoptosis . In the presence of DB01277 , both TNFalpha-induced and obtustatin-induced degeneration of neuronal processes were effectively inhibited . Furthermore , TNFalpha-mediated neuronal degeneration correlated with decreased attachment of PC12/IGF-1R cells to collagen IV and with a reduced level of alpha1beta1 integrin , consistent with a role for this surface protein in the maintenance of neuronal processes . Thus the neuroprotective effects of DB01277 are not restricted to its antiapoptotic properties but also involve an additional neuroprotective mechanism , by which DB01277 counteracts the negative effect of TNFalpha on alpha1beta1 integrin-mediated attachment to collagen IV . The effect of rabbit antithymocyte globulin on human DB05914 . Mesenchymal stem cells ( MSCs ) possess immunomodulatory properties which are of key interest for their application in autoimmunity and transplantation . In transplantation , administration of MSCs has shown promising results in preclinical models and has recently moved to clinical trials . Therefore , it is important to study the interactions between MSCs and immunosuppressive drugs currently used in transplantation . We aimed to analyze the effect of rabbit antithymocyte globulin ( DB00098 ) MSCs . MSCs were obtained from perirenal fat of kidney donors and exposed to ranging doses of DB00098 ( DB00098 (®) , Genzyme ; 0.5-100 μg/ml ) . Binding of DB00098 , effects on viability and susceptibility to be killed by cytotoxic lymphocytes as well as effects on their immunosuppressive potential of MSCs were tested . DB00098 binds dose-dependently to MSCs . This binding was associated with slightly impaired viability after 48 and 72 h when compared with nonexposed MSCs . In contrast to nontreated MSCs , DB00098 preexposed MSCs were susceptible to be lysed by cytokine-activated CD8(+) cytotoxic cells and NKT cells . The capacity of MSCs to suppress the proliferation of anti-CD3/ P10747 activated P01730 and CD8 T cells were reduced by the presence of DB00098 in the culture . DB00098 reduces the viability and antiproliferative capacity of MSCs in a dose-dependent manner and converts them into targets for CD8 T cells and NKT cell lysis . Linkage analysis of multiple sclerosis with candidate region markers in Sardinian and Continental Italian families . Previous genome screens in multiple sclerosis have shown some evidence of linkage in scattered chromosomal regions . Although in no case the evidence of each single study was compelling and although in general the linkage ' peaks ' of the different studies did not coincide , some regions can be considered likely candidates for the presence of MS risk genes because of the clustering of MLS scores and homology with eae loci . We performed a linkage analysis of markers in these regions and of intragenic markers of some individual candidate genes ( HLA- Q8IUH3 , P16410 , P15248 , P02649 , P10415 , P20333 ) . For the first time , Southern European populations were targeted , namely Continental Italians and Sardinians . A total of 69 multiplex families were typed for 67 markers by a semi-automatic fluorescence-based assay . Results were analysed for linkage by two non-parametric tests : GENEHUNTER and SimIBD . In general , the linkage scores obtained were low , confirming the conclusion that no gene is playing a major role in the disease . However , some markers , in 2p11 , 3q21.1 , 7p15.2 and 22q13.1 stood out as promising since they showed higher scores with one or the other test . This stimulates further association analysis of a large number of simplex families from the same populations . 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 . The effect of low-dose estroprogestinic preparations on prothrombin complex factors : no significant increase after an 8-month trial . The behavior of the prothrombin complex factors in 16 healthy women during low-dose estroprogestinic treatment ( laevonorgestrel 0.15 mg and ethynilestradiol 0.30 mg ) at basal conditions and during 8 months of therapy has been investigated . We found a statistically significant decrease of the PTT ( Partial P13726 Time ) . The prothrombin time , on the other hand , became slightly decreased , but not to a statistically significant extent . Among the prothrombin time derived tests for evaluating the prothrombin complex only the PP test ( P00734 P08709 test ) was significantly shortened . Of the coagulation factors ( factors II , VII and X ) only a modest , but not statistically significant , increase in Factor VII and Factor X was noted . We conclude that , during the 8 month observation period , prothrombin complex factors are not altered substantially . Angiotensin converting enzyme inhibitor attenuates oxidative stress-induced endothelial cell apoptosis via p38 Q96HU1 kinase inhibition . BACKGROUND : The effects of angiotensin converting enzyme ( P12821 ) inhibitors on oxidative stress-induced apoptosis of endothelial cells and the intracellular signaling were investigated . METHODS : Cultured endothelial cells derived from a bovine carotid artery were treated with H2O2 or P01375 to induce apoptosis . Apoptosis was evaluated by DNA fragmentation and cell viability , p38 Q96HU1 kinase activity by Western blotting , and oxidative stress by formation of 8-isoprostane . The effects of P12821 inhibitors were examined by adding them into the medium throughout the experiments . RESULTS : Apoptosis was attenuated by P12821 inhibitors , temocapril and captopril , in a dose-dependent manner ( 1-100 micromol/l ) . H2O2 ( 0.2 mmol/l for 1.5 h ) or P01375 ( 10 ng/ml for 72 h ) treatment stimulated the activities of p38 Q96HU1 kinase . DB08836 and captopril decreased the activity of p38 Q96HU1 kinase as well as 8-isoprostane formation induced by H2O2 . A p38 Q96HU1 kinase inhibitor , SB203580 , partially inhibited the effect of temocapril on apoptosis . CONCLUSIONS : These results suggest that P12821 inhibitors protect endothelial cells from oxidative stress-induced apoptosis , and that p38 Q96HU1 kinase plays a critical role in the process . 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 . 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 . HER-2 peptides p776 and P08709 , N-terminal-linked with Ii-Key tetramer ( LRMK ) help the proliferation of E75-TCR+ cells : The dependency of help on the side chains of LRMK-extended peptide pointed towards the T cell receptor . The objective of this study was to determine whether peptides consisting of the Ii-Key peptide LRMK linked to the N-terminal ends of HER-2 peptides would stimulate the expansion of antigen-specific E75-TCR+CD8+ cells . The peptides tested were N-acetylated and linked to an alpha-amide at the C-terminus ; some of the peptides contained epsilon-aminovaleric acid ( Ava ) between the LRMK and the HER-2 peptide . Of the seven LRMK-HER-2 peptides tested to date , three effectively induced P01579 production by peripheral blood mononuclear cells ( PBMCs ) from healthy donors and women with ductal carcinoma in situ . A fusion peptide , LRMK-Ava-HER-2(777-789) , was more immunogenic than the natural HER-2(777-789) antigen , G89 , with regard to P01579 production . In combination with the CD8-activating peptide E75 [ HER-2(369-377) ] LRMK-p776 and LRMK-Ava- P08709 induced the proliferation of E75-TCR(Med+Hi) CD8+ cells to a greater extent than did 1,000 or 5,000 nM of E75 alone , respectively . The induction effects were strongest at 600 nM for LRMK-p776 and 3,000 nM for LRMK-Ava- P08709 . At 3,000 nM , LRMK-p776 was cytotoxic to PBMCs . LRMK-p776 and P08709 had a similar specificity and preferences for binding HLA-DR molecules . The molecular modeling of HLA-DR:LRMK-p776 and HLA-DR:LRMK-Ava- P08709 complexes revealed the side chains of the peptides , which pointed towards the T-cell receptor . Differences in side chain orientation introduced by various N-terminal extensions of MHC class II-bound peptides should be important for directing P01730 + cells to stimulate CD8+ cells or for eliminating regulatory T cells in cancer immunotherapy . DB01079 , a Q13639 receptor partial agonist , decreases sensitivity to rectal distension in healthy subjects . BACKGROUND : DB01079 reduces the symptoms associated with irritable bowel syndrome , and anti-nociceptive effects have been demonstrated in animals . Its effect on the rectal sensitivity in humans has not been delineated clearly . AIM : To evaluate the action of tegaserod on rectal sensitivity in response to distension by means of a reflexological technique based on electrophysiological recordings of the RIII nociceptive reflex . METHODS : A randomized , double-blind , placebo-controlled study , performed in 20 healthy women , quantified the effects of slow or rapid rectal distensions on the RIII reflex at baseline and on day 8 following treatment with either placebo or tegaserod ( 6 mg b.d. ) . RESULTS : At baseline , slow distensions performed up to the pain threshold induced gradual inhibitions of the RIII reflex . On day 8 , these inhibitory effects were significantly reduced in the tegaserod group , but not in the placebo group ( P = 0.0001 ) . The effects of rapid distensions were not significantly modified by tegaserod or placebo . The intensity of subjective pain perception and rectal compliance were not altered by either treatment . CONCLUSION : These results suggest that tegaserod reduces the sensitivity to rectal distension in healthy subjects and interacts with the processing of sensory visceral information . Discovery and role of methylidene imidazolone , a highly electrophilic prosthetic group . The elimination of ammonia from alpha-amino acids is a chemically difficult process . While the non-acidic beta-proton has to be abstracted , the much more acidic ammonium protons must remain untouched to maintain the leaving group ability of this positively charged group . DB00117 and phenylalanine ammonia-lyases ( P42357 and Q9P2V4 ) possess a catalytically essential electrophilic group which has been believed to be dehydroalanine for 30 years . Recently , the X-ray structure of P42357 has been solved . The electron density was not consistent with dehydroalanine but showed the presence of methylidene imidazolone ( Q9NP71 ) instead . The high electrophilicity of this prosthetic group as well as the geometry at the active site support a previously proposed mechanism involving a Friedel-Crafts-type attack at the aromatic ring of the substrate . Further biochemical evidence for this unprecedented electrophile-assisted ammonia elimination is also presented . Although no X-ray structure of Q9P2V4 has been published as yet , spectrophotometrical evidence for the presence of Q9NP71 has been provided . Finally , a chemical model for the Q9P2V4 reaction is described .

MH_train_1581

MH_train_1581

MH_train_1581

interacts_with DB01109?

multiple_choice

Molecular response of HL-60 cells to mitotic inhibitors vincristine and taxol visualized with apoptosis-related gene expressions , including the new member Q9HB09 . DB01229 and vincristine belong to a group of anticancer drugs that target microtubules , subsequently arresting cells at the mitotic phase of the cell cycle and inducing programmed cell death . The P10415 ( bcl-2 ) family of genes is of known implication in apoptosis induced by various stimuli , among which Q9HB09 , a new member of the family , cloned by our group . For further insights into the mechanisms and molecular targets implicated and modified as a result of apoptosis induced by these two mitosis-arresting drugs , we studied the possible alterations , at the mRNA level , of various apoptosis-related genes ( P10415 , Q07812 , Q9HB09 , CASPASE-3 , FAS ) after leukemia cell ( HL-60 ) treatment with these drugs . The kinetics of cell toxicity were evaluated by the MTT [ 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ] method , trypan blue staining , and cell proliferation efficiency ; apoptosis induction was assayed by endonucleosomal cleavage of DNA ( DNA laddering ) ; and the expression levels of the genes were analysed by RT-PCR , using gene-specific primers . The percentage of nonviable cells was upregulated with increasing cell exposure time and drug concentrations to both taxol and vincristine . Distinct modulations of apoptosis-related genes at the mRNA level were also observed , mainly concerning P10415 and Q9HB09 along apoptosis induction . Our results indicate and support the hypothesis that the apoptosis-related genes P10415 and Q9HB09 respond similarly to treatment of the human , acute , myelocytic leukemia HL60 cells with the anticancer drugs vincristine and taxol though in a drug-specific and time-dependent manner . 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 . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . 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 . 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 . [ Plasma exchange with very low molecular weight heparin CY 222. Biological profile and therapeutic value ] . In a clinical , between-patient study we investigated the effects of a VLMW DB01109 fragment ( CY 222 ) versus standard heparin ( SH ) in plasma exchanges ( n = 10 ) on coagulation factors ( CF ) . DB09222 ( FGN ) , II , V , VIIF + X , IX , XI , XII , VIIIc , VIIIRag , VIIIvwf and P01008 , ProtC , ProtS , P00747 ( Q9UQ90 ) , Activated P13726 time ( APTT ) , P00734 time ( PT ) , Thrombin time ( TT ) , anti factor Xa ( Axa ) , DDimer , Platelet count . DB01109 was administered as a bolus and by infusion during the session , CY 222 as a bolus dose only ; 1 to 1.5 plasma volume was exchanged with substitution by 5 % albumin . Results ( mean , s.d. ) at the end of the session ( End ) and 4 hours later ( DB00451 ) were analyzed and showed no differences between groups ( with CY 222 and with SH ) for technical and clinical findings . Biologically , CF were similar in both groups except for Factor XII levels at the end of the session , and Factors II , V , XII at DB00451 . Prolonged APTT in all samples appear related to low FGN . Significant differences in Axa activities were found for each treatment when compared with its own standard , suggesting different ranges of activities of both drugs . Changes in D-dimer levels differed during the session and four hours after the session with the drug tested , and could be related to their mode of administration . Clinical efficiency and tolerance were excellent both with CY 222 and SH . [ Activation of coagulation cascade in children during an idiopathic nephrotic syndrome relapse ] . The objective of this study was to assess concentrations of selected markers of coagulation in children with relapse of idiopathic nephrotic syndrome during a 6-week therapy . Study groups : 22 subjects ( 32 relapses ) -- 14 males , 8 females ( mean age 7.15 +/- 1.5 y. ) with no thrombotic complications were included into the study . All children were clinically steroid-sensitive . METHODS : Coagulation markers ( platelet count , thrombin time , APTT , INR , fibrinogen 1 + 2 fragments ( F1 + 2 ) , thrombin-antithrombin complexes ( TAT ) , serum levels of D-dimer ( DD ) , fibrin monomers ( FM ) and antithrombin activity ( P01008 ) ) were measured three times : on admission , after 2 and 6 weeks . The control group consisted of 13 healthy children . RESULTS : Serum concentration of TAT or F1 + 2 did not differ between 3 stages ( p > 0.05 ) . However , values at 0 and 2 weeks were significantly higher than in control group ( p < 0.05 ) . We found no correlation between TAT or F1 + 2 and FBG , ALB , TCH , TG levels . [ table : see text ] CONCLUSIONS : The coagulation cascade in relapse of NS was activated during first 6 weeks of therapy whereas metabolic disturbances ( low ALB , high P02675 , TCH , TG , high platelets ) normalized . It is speculative whether it was caused by active immunological process but definitely it resulted in " prothrombotic state " in P01308 patients . Molecular characterization of pediatric gastrointestinal stromal tumors . PURPOSE : Pediatric gastrointestinal stromal tumors ( GIST ) are rare and occur preferentially in females as multifocal gastric tumors , typically lacking mutations in P10721 and P16234 . As P10721 oncoprotein is consistently overexpressed in pediatric GIST , we sought to investigate the activation of P10721 downstream targets and alterations of P10721 / P16234 gene copy number , mine novel therapeutic targets by gene expression , and test tyrosine kinase receptor activation by proteomic profiling . EXPERIMENTAL DESIGN : Seventeen pediatric GISTs were investigated for P10721 / P16234 genotype and biochemical activation of P10721 downstream targets . The transcriptional profile of 13 nodules from 8 pediatric patients was compared with 8 adult wild-type ( WT ) GISTs , including 3 young adults . The drug sensitivity of second-generation kinase inhibitors was tested in murine Ba/ P13726 cells expressing human WT P10721 , as well as in short-term culture of explants of WT GIST cells . RESULTS : A P10721 / P16234 WT genotype was identified in all 12 female patients , whereas two of five males had either a P10721 exon 11 or P16234 exon 18 mutation . P10721 downstream targets were consistently activated . Pediatric GISTs showed a distinct transcriptional signature , with overexpression of Q8WXS3 , Q6DJT9 , P08069 , P08620 , and Q92832 . In vitro studies showed that nilotinib , sunitinib , dasatinib , and sorafenib are more effective than imatinib against WT P10721 . CONCLUSIONS : Rare cases of pediatric GIST may occur in male patients and harbor activating P10721 / P16234 mutations . Pediatric GISTs show distinct transcriptional signature , suggesting a different biology than WT GIST in adults . In vitro drug screening showed that second-generation kinase inhibitors may provide greater clinical benefit in pediatric GIST . The α7 nicotinic acetylcholine receptor and the acute stress response : maternal genotype determines offspring phenotype . α7 Nicotinic acetylcholine receptors ( α7nAchRs ) modulate immune activation by suppressing production of pro-inflammatory cytokines in peripheral immune cells . α7nAchRs also modulate inhibitory output in the hippocampus , which provides input to key circuits of the Q9Y251 axis . Therefore , the α7 nicotinic acetylcholine receptor gene ( P36544 ) may be associated with cortisol stress response . Polymorphisms in the P36544 promoter decrease its expression and may dampen the cholinergic response , leading to an increase in inflammation . Increased inflammation may change the intrauterine environment , altering neuroendocrine development in the offspring . Maternal P36544 genotype could affect an offspring 's Q9Y251 regulation via reprogramming in utero . Patients with allergic disorders have a differential cortisol response to stress . This study utilized samples collected from a cohort of 198 adolescents in a previous study of atopic disorders , who demonstrated a disturbance in Q9Y251 response associated with atopy . Salivary cortisol samples collected from the adolescents after a series of laboratory procedures and DNA samples collected from the adolescents and their parents were used for further analysis . DNA samples were genotyped for allelic variation in the P36544 promoter . Genetic association analyses with the cortisol levels were performed in the adolescents . Maternal genotype influences were investigated for the P36544 gene . We also included maternal and child atopy diagnosis as covariates in determining cortisol levels and tested for association of P36544 to atopy . Polymorphisms in the P36544 promoter were associated with lower cortisol levels after a small laboratory stress . Our findings also show that although the child 's P36544 genotype affects stress response , the maternal genotype has a stronger influence on cortisol release after stress in male offspring . These effects were independent of atopy status . DB01819 cycling via mitochondrial phosphoenolpyruvate carboxykinase links anaplerosis and mitochondrial GTP with insulin secretion . Pancreatic beta-cells couple the oxidation of glucose to the secretion of insulin . Apart from the canonical K( DB00171 )-dependent glucose-stimulated insulin secretion ( GSIS ) , there are important K( DB00171 )-independent mechanisms involving both anaplerosis and mitochondrial GTP ( mtGTP ) . How mtGTP that is trapped within the mitochondrial matrix regulates the cytosolic calcium increases that drive GSIS remains a mystery . Here we have investigated whether the mitochondrial isoform of phosphoenolpyruvate carboxykinase ( Q16822 ) is the GTPase linking hydrolysis of mtGTP made by succinyl- DB01992 synthetase ( SCS-GTP ) to an anaplerotic pathway producing phosphoenolpyruvate ( PEP ) . Although cytosolic PEPCK ( P35558 ) is absent , Q16822 message and protein were detected in P01308 -1 832/13 cells , rat islets , and mouse islets . PEPCK enzymatic activity is half that of primary hepatocytes and is localized exclusively to the mitochondria . Novel (13)C-labeling strategies in P01308 -1 832/13 cells and islets measured substantial contribution of Q16822 to the synthesis of PEP . As high as 30 % of PEP in P01308 -1 832/13 cells and 41 % of PEP in rat islets came from Q16822 . The contribution of Q16822 to overall PEP synthesis more than tripled with glucose stimulation . Silencing the Q16822 gene completely inhibited GSIS underscoring its central role in mitochondrial metabolism-mediated insulin secretion . Given that mtGTP synthesized by SCS-GTP is an indicator of TCA flux that is crucial for GSIS , Q16822 is a strong candidate to link mtGTP synthesis with insulin release through anaplerotic PEP cycling . The attenuation of experimental lung metastasis by a bile acid acylated-heparin derivative . The inhibitory efficacies of new bile acid acylated-heparin derivative ( heparin-DOCA ) were evaluated on experimental lung metastasis . We evaluated the effect of heparin-DOCA on intercellular interactions including those between B16F10 and thrombin-activated platelets and P01375 -activated HUVECs , and between B16F10 and immobilized mouse P16109 . In addition , the inhibitory effects of heparin-DOCA on adhesion and invasion of B16F10 to Matrigel were studied . In an animal mouse study , the blood clot formation and the retention of red fluorescence protein ( RFP ) -B16F10 in lungs were assessed after heparin-DOCA and RFP-B16F10 intravenous administration . Furthermore , we investigated the anti-metastatic effect of heparin-DOCA against lung metastasis induced by B16F10 and SCC7 . DB01109 -DOCA inhibited intercellular interactions between B16F10 and activated platelets or activated HUVECs by blocking P- and P16581 -mediated interactions . Moreover , it reduced adhesion and invasion of B16F10 to Q13201 , thereby affecting the reduction of early retention of B16F10 in the lung . DB01109 -DOCA attenuated lung colony formation on the surfaces and in interior of the lung , and attenuated metastasis by B16F10 and SCC7 . These results suggest that heparin-DOCA may have potentials as therapeutic agent that prevents tumor metastasis and progression . [ 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 . P08069 targeted therapeutics : novel compounds and novel treatment strategies for cancer medicine . The insulin-like growth factor 1 receptor ( IGF-1R ) and its associated signalling system has provoked considerable interest over recent years as a novel therapeutic target in cancer . A brief outline of the IGF-1R signalling system and the rationale for its use in cancer medicine is given . This is followed by a discussion of the different possible targets within the IGF-1R system , and drugs developed to interact at each target . A systems-based approach is then used to review the in vitro and in vivo data in the published literature of the following compounds targeting IGF-1R components using specific examples : growth hormone releasing hormone antagonists ( e.g. JV-1-38 ) , growth hormone receptor antagonists ( e.g. pegvisomant ) , IGF-1R antibodies ( e.g. CP-751,871 , AVE1642/EM164 , DB05759 , P35240 -717454 , BIIB022 , Q99217 479 , MK-0646/h7C10 ) , and IGF-1R tyrosine kinase inhibitors ( e.g. BMS-536942 , BMS-554417 , DB00238 -AEW541 , DB00238 -ADW742 , AG1024 , potent quinolinyl-derived imidazo (1,5-a)pyrazine PQIP , picropodophyllin PPP , Nordihydroguaiaretic acid Insm-18/NDGA ) . The following tumour types are specifically discussed : lung , breast , colorectal , pancreatic , NETs , sarcoma , prostate , leukaemia , multiple myeloma . Other tumour types are mentioned briefly : squamous cell carcinoma of the head and neck , melanoma , glioblastoma , ovary , gastric and mesothelioma . Results of early stage clinical trials , involving recently patented drugs. are included where appropriate . We then outline the current understanding of toxicity related to IGF-1R targeted therapy , and finally outline areas for further research . P16109 - and heparanase-dependent antimetastatic activity of non-anticoagulant heparins . Vascular cell adhesion molecules , P- and L-selectins , facilitate metastasis of cancer cells in mice by mediating interactions with platelets , endothelium , and leukocytes . Q9Y251 is an endoglycosidase that degrades heparan sulfate of extracellular matrix , thereby promoting tumor invasion and metastasis . DB01109 is known to efficiently attenuate metastasis in different tumor models . Here we identified modified , nonanticoagulant species of heparin that specifically inhibit selectin-mediated cell-cell interactions , heparanase enzymatic activity , or both . We show that selective inhibition of selectin interactions or heparanase with specific heparin derivatives in mouse models of MC-38 colon carcinoma and B16-BL6 melanoma attenuates metastasis . Selectin-specific heparin derivatives attenuated metastasis of MC-38 carcinoma , but heparanase-specific derivatives had no effect , in accordance with the virtual absence of heparanase activity in these cells . DB01109 derivatives had no further effect on metastasis in mice deficient in P- and P14151 , indicating that selectins are the primary targets of heparin antimetastatic activity . Selectin-specific and heparanase-specific derivatives attenuated metastasis of B16-BL6 melanomas to a similar extent . When mice were injected with a derivative containing both heparanase and selectin inhibitory activity , no additional attenuation of metastasis could be observed . Thus , selectin-specific heparin derivatives efficiently attenuated metastasis of both tumor cell types whereas inhibition of heparanase led to reduction of metastasis only in tumor cells producing heparanase . NO overproduction by P29474 precedes hyperdynamic splanchnic circulation in portal hypertensive rats . Chronic high blood flow and the hyperdynamic circulatory syndrome in portal hypertension are associated with endothelial constitutive nitric oxide ( NO ) synthase ( P29474 ) upregulation and increased NO release . In portal vein-ligated ( PVL ) rats the splanchnic circulation is not yet hyperdynamic on day 3 postoperatively . In vitro perfused superior mesenteric arteries ( SMAs ) of day 3 PVL and sham rats were challenged with increasing flow rates or the alpha-adrenoreceptor agonist methoxamine ( 30 and 100 microM ) before and after incubation with the NO inhibitor , Nomega-nitro-L-arginine ( DB04223 , 10(-4) M ) . Perfusate NO metabolite ( NOx ) concentrations were measured by chemiluminescence . PVL rats expressed a significant hyporesponsiveness to increases in flow rate or methoxamine that was overcome by incubation with DB04223 . The PVL vasculature showed significantly higher slopes of NOx production vs. flow-induced shear stress , higher increases in perfusate NOx concentration in response to methoxamine , and higher P29474 protein levels ( Western blot ) compared with sham rats . In conclusion , P29474 -upregulation and increased NO release by the SMA endothelium occur before the development of the hyperdynamic splanchnic circulation , suggesting a primary role of NO in the pathogenesis of arterial vasodilatation . Endothelial protective genes induced by statin are mimicked by Q13164 activation as triggered by a drug combination of FTI-277 and GGTI-298 . BACKGROUND : Statins are potent inhibitors of cholesterol biosynthesis and are clinically beneficial in preventing cardiovascular diseases , however , the therapeutic utility of these drugs is limited by myotoxicity . Here , we explored the mechanism of statin-mediated activation of Q13164 in the human endothelium with the goal of identifying compounds that confer endothelial protection but are nontoxic to muscle . METHODS : An Q13164 -one hybrid luciferase reporter transfected into COS-7 cells with pharmacological and molecular manipulations dissected the signaling pathway leading to statin activation of Q13164 . qRT-PCR of HUVEC cells documented the transcriptional activation of endothelial-protective genes . Lastly , morphological and cellular DB00171 analysis , and induction of atrogin-1 in C2C12 myotubes were used to assess statin-induced myopathy . RESULTS : Statin activation of Q13164 is dependent on the cellular reduction of GGPPs . Furthermore , we found that the combination of FTI-277 ( inhibitor of farnesyl transferase ) and GGTI-298 ( inhibitor of geranylgeranyl transferase I ) mimicked the statin-mediated activation of Q13164 . FTI-277 and GGTI-298 together recapitulated the beneficial effects of statins by transcriptionally upregulating anti-inflammatory mediators such as P29474 , P07204 , and Q9Y5W3 . Finally , C2C12 skeletal myotubes treated with both FTI-277 and GGTI-298 evoked less morphological and cellular changes recognized as biomarkers of statin-associated myopathy . CONCLUSIONS : Statin-induced endothelial protection and myopathy are mediated by distinct metabolic intermediates and co-inhibition of farnesyl transferase and geranylgeranyl transferase I confer endothelial protection without myopathy . GENERAL SIGNIFICANCE : The combinatorial FTI-277 and GGTI-298 drug regimen provides a promising alternative avenue for endothelial protection without myopathy . 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 . Inhibition of a thrombin anion-binding exosite-2 mutant by the glycosaminoglycan-dependent serpins protein C inhibitor and heparin cofactor II . Antithrombin ( P01008 ) , heparin cofactor II ( HCII ) and protein C inhibitor ( P05154 ; also named plasminogen activator inhibitor-3 ) are serine protease inhibitors ( serpins ) whose thrombin inhibition activity is accelerated in the presence of glycosaminoglycans . We compared the inhibition properties of P05154 and HCII to P01008 using R93A/R97A/R101A thrombin , an anion-binding exosite-2 ( exosite-2 ) mutant that has greatly reduced heparin-binding properties . DB01109 -enhanced P05154 inhibition of R93A/R97A/R101A thrombin was only approximately 2-fold compared to 40-fold enhancement with wild-type recombinant thrombin . P07204 ( TM ) ( with or without the chondroitin sulfate moiety ) accelerated P05154 inhibition of both wild-type and R93A/R97A/R101A thrombins . HCII achieved the same maximum activity in the presence of heparin with both wild-type and R93A/R97A/R101A thrombins ; however , the optimum heparin concentration was 20 times greater than the reaction with wild-type thrombin , indicative of a decrease in heparin affinity . Dermatan sulfate ( DSO4 ) -catalyzed HCII thrombin inhibition was unchanged in R93A/R97A/R101A thrombin compared to wild-type recombinant thrombin . These results suggest that P05154 is similar to P01008 and depends upon ternary complex formation with heparin and these specific thrombin exosite-2 residues to accelerate thrombin inhibition . In contrast , HCII does not require DB00125 (93) , DB00125 (97) and DB00125 (101) of thrombin exosite-2 and further supports the hypothesis that HCII uses an allosteric process following glycosaminoglycan binding to inhibit thrombin . The polyglutamine neurodegenerative protein ataxin 3 regulates aggresome formation . The polyglutamine-containing neurodegenerative protein ataxin 3 ( P01008 ) has deubiquitylating activity and binds ubiquitin chains with a preference for chains of four or more ubiquitins . Here we characterize the deubiquitylating activity of P01008 in vitro and show it trims/edits K48-linked ubiquitin chains . P01008 also edits polyubiquitylated (125)I-lysozyme and decreases its degradation by proteasomes . Cellular studies show that endogenous P01008 colocalizes with aggresomes and preaggresome particles of the misfolded cystic fibrosis transmembrane regulator ( P13569 ) mutant CFTRDeltaF508 and associates with histone deacetylase 6 and dynein , proteins required for aggresome formation and transport of misfolded protein . Small interfering RNA knockdown of P01008 greatly reduces aggresomes formed by CFTRDeltaF508 , demonstrating a critical role of P01008 in this process . Wild-type P01008 restores aggresome formation ; however , P01008 with mutations in the active site or ubiquitin interacting motifs can not restore aggresome formation in P01008 knockdown cells . These same mutations decrease the association of P01008 and dynein . These data indicate that the deubiquitylating activity of P01008 and its ubiquitin interacting motifs as well play essential roles in CFTRDeltaF508 aggresome formation . 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 . P03372 -β ligand treatment modulates dendritic cells in the target organ during autoimmune demyelinating disease . DB00286 act upon nuclear estrogen receptors ( ER ) to ameliorate cell-mediated autoimmune disease . As most immunomodulatory effects of estrogens in EAE have been attributed to the function of ER-α , we previously demonstrated that ER-β ligand treatment reduced disease severity without affecting peripheral cytokine production or levels of CNS inflammation , suggesting a direct neuroprotective effect ; however , the effect of ER-β treatment on the function of immune cells within the target organ remained unknown . Here , we used adoptive transfer studies to show that ER-β ligand treatment was protective in the effector , but not the induction phase of EAE , as shown by decreased clinical disease severity with the preservation of axons and myelin in spinal cords . The analysis of the immune cell infiltrates in the CNS revealed that while ER-β ligand treatment did not reduce overall levels of CNS inflammation , there was a decrease in the DC percentage , and these CNS DC had decreased P01375 -α production . Finally , experiments using DC deficient in ER-β revealed that the expression of ER-β on DC was essential for protective effects of ER-β ligand treatment in EAE . Our results demonstrate for the first time an effect of ER-β ligand treatment in vivo on DC in the target organ of a prototypic cell-mediated autoimmune disease . 8p12 stem cell myeloproliferative disorder : the FOP-fibroblast growth factor receptor 1 fusion protein of the t(6;8) translocation induces cell survival mediated by mitogen-activated protein kinase and phosphatidylinositol 3-kinase/Akt/ P42345 pathways . The FOP-fibroblast growth factor receptor 1 ( P11362 ) fusion protein is expressed as a consequence of a t(6;8) ( q27;p12 ) translocation associated with a stem cell myeloproliferative disorder with lymphoma , myeloid hyperplasia and eosinophilia . In the present report , we show that the fusion of the leucine-rich N-terminal region of FOP to the catalytic domain of P11362 results in conversion of murine hematopoietic cell line Ba/ P13726 to factor-independent cell survival via an antiapoptotic effect . This survival effect is dependent upon the constitutive tyrosine phosphorylation of FOP- P11362 . Phosphorylation of P42224 and of P40763 , but not P42229 , is observed in cells expressing FOP- P11362 . The survival function of FOP- P11362 is abrogated by mutation of the phospholipase C gamma binding site . Mitogen-activated protein kinase ( MAPK ) is also activated in FOP- P11362 -expressing cells and confers cytokine-independent survival to hematopoietic cells . These results demonstrate that FOP- P11362 is capable of protecting cells from apoptosis by using the same effectors as the wild-type P11362 . Furthermore , we show that FOP- P11362 phosphorylates phosphatidylinositol 3 ( P19957 ) -kinase and AKT and that specific inhibitors of P19957 -kinase impair its ability to promote cell survival . In addition , FOP- P11362 -expressing cells show constitutive phosphorylation of the positive regulator of translation p70S6 kinase ; this phosphorylation is inhibited by P19957 -kinase and P42345 ( mammalian target of rapamycin ) inhibitors . These results indicate that translation control is important to mediate the cell survival effect induced by FOP- P11362 . Finally , FOP- P11362 protects cells from apoptosis by survival signals including P10415 overexpression and inactivation of caspase-9 activity . Elucidation of signaling events downstream of FOP- P11362 constitutive activation provides insight into the mechanism of leukemogenesis mediated by this oncogenic fusion protein .

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interacts_with DB00834?

multiple_choice

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 . P04150 and sequential P04637 activation by dexamethasone mediates apoptosis and cell cycle arrest of osteoblastic MC3T3-E1 cells . Glucocorticoids play a pivotal role in the proliferation of osteoblasts , but the underlying mechanism has not been successfully elucidated . In this report , we have investigated the molecular mechanism which elucidates the inhibitory effects of dexamethasone on murine osteoblastic MC3T3-E1 cells . It was found that the inhibitory effects were largely attributed to apoptosis and P55008 phase arrest . Both the cell cycle arrest and apoptosis were dependent on glucocorticoid receptor ( GR ) , as they were abolished by GR blocker DB00834 pre-treatment and GR interference . P55008 phase arrest and apoptosis were accompanied with a p53-dependent up-regulation of P38936 and pro-apoptotic genes Q13794 and PUMA . We also proved that dexamethasone ca n't induce apoptosis and cell cycle arrest when p53 was inhibited by p53 RNA interference . These data demonstrate that proliferation of MC3T3-E1 cell was significantly and directly inhibited by dexamethasone treatment via aberrant GR activation and subsequently P04637 activation . Targeted deletion of cannabinoid receptors P21554 and CB2 produced enhanced inflammatory responses to influenza A/PR/8/34 in the absence and presence of Delta9-tetrahydrocannabinol . We have previously reported that Delta-9-tetrahydrocannabinol ( Delta(9)-THC ) -treated mice challenged with influenza virus A/PR/8/34 ( PR8 ) developed increased viral hemagglutinin 1 ( H1 ) mRNA levels and decreased monocyte and lymphocyte recruitment to the pulmonary airways when compared with mice challenged with PR8 alone . The objective of the present study was to examine the role of cannabinoid ( CB(1)/CB(2) ) receptors in mediating the effects of Delta(9)-THC on immune and epithelial cell responses to PR8 . In the current study , Delta(9)-THC-treated CB(1)/CB(2) receptor null ( CB(1)-/-/CB(2)-/- ) and wild-type mice infected with PR8 had marked increases in viral H1 mRNA when compared with CB(1)-/-/CB(2)-/- and wild-type mice challenged with PR8 alone . However , the magnitude of the H1 mRNA levels was greatly reduced in CB(1)-/-/CB(2)-/- mice as compared with wild-type mice . In addition , Delta(9)-THC-treated CB(1)-/-/CB(2)-/- mice infected with PR8 had increased P01730 + T cells and P01579 in bronchoalveolar lavage fluid with greater pulmonary inflammation when compared with Delta(9)-THC-treated wild-type mice infected with PR8 . Delta(9)-THC treatment of CB(1)-/-/CB(2)-/- mice in the presence or absence of PR8 challenge also developed greater amounts of mucous cell metaplasia in the affected bronchiolar epithelium . Collectively , the immune and airway epithelial cell responses to PR8 challenge in Delta(9)-THC-treated CB(1)-/-/CB(2)-/- and wild-type mice indicated the involvement of CB(1)/CB(2) receptor-dependent and -independent mechanisms . Gallic acid inhibits migration and invasion of SCC-4 human oral cancer cells through actions of NF-κB , Ras and matrix metalloproteinase-2 and -9 . Oral cancer is one of the major causes of mortality in humans and squamous cell carcinoma is the most common type of oral cancer . Gallic acid ( GA ) is a natural product that induces cell death through cell cycle arrest and induction of apoptosis . There is no available information on whether GA affects cell migration and invasion of human oral cancer cells . We determined if GA inhibited migration and invasion of SCC-4 ( human squamous cell carcinoma ) human oral cancer cells . GA significantly inhibited migration and invasion of SCC-4 cells based on results from the wound healing assay and Matrigel Cell Migration Assay and Invasion System . We also showed that GA significantly inhibited matrix metalloproteinase ( MMP ) -2 and P14780 activity . GA reduced protein levels of Q05397 , Q99759 , p- Q9NZJ5 , p-p38 , p- P45983 /2 , p- P27361 /2 , Q07889 , RhoA , Ras , PKC , p-AKT(Thr308) , PI3K , NF-κB p65 , P08253 and P14780 in SCC-4 cells . Translocation of NF-κB and RhoA from the cytosol to the nucleus was reduced by GA in SCC-4 cells . In summary , GA inhibits migration and invasion of SCC-4 cells by inhibiting NF-κB expression causing suppression of P08253 and P14780 activity . GA may have potential as a therapeutic agent for the treatment of oral cancer . Pixelation effect removal from fiber bundle probe based optical coherence tomography imaging . A method of eliminating pixelization effect from en face optical coherence tomography ( O75051 ) image when a fiber bundle is used as an O75051 imaging probe is presented . We have demonstrated that applying a histogram equalization process before performing a weighted-averaged Gaussian smoothing filter to the original lower gray level intensity based image not only removes the structural artifact of the bundle but also enhances the image quality with minimum blurring of object 's image features . The measured contrast-to-noise ratio ( P21554 ) for an image of the US DB09337 Force test target was 14.7dB ( 4.9dB ) , after ( before ) image processing . In addition , by performing the spatial frequency analysis based on two-dimensional discrete Fourier transform ( 2-D DFT ) , we were able to observe that the periodic intensity peaks induced by the regularly arrayed structure of the fiber bundle can be efficiently suppressed by 41.0dB for the first nearby side lobe as well as to obtain the precise physical spacing information of the fiber grid . The proposed combined method can also be used as a straight forward image processing tool for any imaging system utilizing fiber bundle as a high-resolution imager . 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 . 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 . Interaction between fatty acid synthase- and ErbB-systems in ovarian cancer cells . P49327 ( P49327 ) represents a metabolic oncogene . It produces phospholipids for membrane microdomains that accommodate receptor tyrosine kinases including Epidermal Growth Factor-Receptor ( P00533 , ErbB1 ) and ErbB2 ( P04626 /neu ) . P49327 and ErbBs are overexpressed in ovarian cancer . We examined the effect of P49327 and ErbB inhibition on A2780 and SKOV3 ovarian cancer cells . Growth assays reveal that P49327 inhibitor C75 sensitizes tumor cells against anti-ErbB drugs ( pelitinib [ Q9Y259 -569 ] , canertinib [ DB05424 ] , erlotinib , cetuximab , matuzumab , trastuzumab ) suggesting P49327 /ErbB cooperation . qRT-PCR and Western blotting revealed that C75 represses P49327 , P00533 , ErbB2 , and AKT suggesting that P49327 -induced membrane microdomains accommodate/stabilize ErbBs and facilitate AKT recruitment/activation . Our data indicate that AKT is crucial for ErbB/ P49327 interaction , AKT cross-inhibits P29323 and feeds loops that boost P49327 and P00533 transcription , and P00533 and ErbB2 must be co-silenced for maximal P49327 downregulation . Taken together , interference with P49327 and ErbB abrogates their oncogenicity and should be exploited for ovarian cancer treatment . 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 . c-Src modulates estrogen-induced stress and apoptosis in estrogen-deprived breast cancer cells . The emergence of anti-estrogen resistance in breast cancer is an important clinical phenomenon affecting long-term survival in this disease . Identifying factors that convey cell survival in this setting may guide improvements in treatment . Estrogen ( E2 ) can induce apoptosis in breast cancer cells that have been selected for survival after E2 deprivation for long periods ( MCF-7:5C cells ) , but the mechanisms underlying E2-induced stress in this setting have not been elucidated . Here , we report that the c-Src kinase functions as a key adapter protein for the estrogen receptor ( ER , P03372 ) in its activation of stress responses induced by E2 in MCF-7:5C cells . E2 elevated phosphorylation of c-Src , which was blocked by 4-hydroxytamoxifen ( DB04468 ) , suggesting that E2 activated c-Src through the ER . We found that E2 activated the sensors of the unfolded protein response ( UPR ) , IRE1α ( O75460 ) and Q9NZJ5 kinase ( Q9NZJ5 ) , the latter of which phosphorylates eukaryotic translation initiation factor-2α ( eIF2α ) . E2 also dramatically increased reactive oxygen species production and upregulated expression of heme oxygenase P09601 ( P09601 ) , an indicator of oxidative stress , along with the central energy sensor kinase AMPK ( P54646 ) . Pharmacologic or RNA interference-mediated inhibition of c-Src abolished the phosphorylation of eIF2α and AMPK , blocked E2-induced ROS production , and inhibited E2-induced apoptosis . Together , our results establish that c-Src kinase mediates stresses generated by E2 in long-term E2-deprived cells that trigger apoptosis . This work offers a mechanistic rationale for a new approach in the treatment of endocrine-resistant breast cancer . P06401 activation of extranuclear signaling pathways in regulating p53 expression in vascular endothelial cells . We previously showed that progesterone ( P4 ) inhibited the proliferation of human umbilical vein endothelial cells ( HUVECs ) through a p53-dependent pathway . Now we investigated further the molecular mechanism underlying the hormone activity . In cultured HUVECs , P4 increased the protein levels of phosphorylated Src ( p-Src ) , P04049 , and P29323 . The levels of p-Src and p-Src-progesterone receptor complex in HUVECs were increased by P4 treatment . These effects were blocked by pretreatment with a progesterone receptor antagonist , DB00834 . The P4-induced increase in p53 transactivity was abolished by pretreatment with Src kinase inhibitors . Moreover , administration with cSrc antisense oligonucleotide prevented the P4-induced increases of the levels of p53 mRNA and protein . These data suggest that P4-induced up-regulation of p53 might be mediated through activation of cSrc . Pretreatment with Src kinase inhibitors also prevented P4-induced membrane translocation of Kras and increases of the protein levels of phosphorylated Raf and phosphorylated P29323 . Transfection with dominant-negative P28482 prevented the P4-induced increases of protein level and promoter activity of p53 and a decrease of thymidine incorporation . P4 also increased nuclear factor-κB ( NF-κB ) nuclear translocation and NF-κB binding onto the p53 promoter . These effects were abolished by pretreatment with P29323 inhibitors . The P4-induced up-regulation of the p53 promoter activity was prevented by preadministration with dominant-negative P28482 or NF-κB inhibitors . Taken together , our data suggest that the cSrc/Kras/ P04049 / P28482 /NF-κB signaling pathway contributes to the P4-induced up-regulation of p53 in HUVECs . These findings highlight progesterone receptor activation of extranuclear signaling pathways in regulating p53 and cell cycle progression in HUVECs . Characterization of P21554 cannabinoid receptors using receptor peptide fragments and site-directed antibodies . The mechanism by which P21554 cannabinoid receptors are coupled to the Gi/Go class of G proteins was studied . A peptide representing the juxtamembrane carboxyl terminus robustly stimulated guanosine-5'-O-(3-thio)triphosphate binding . Peptides simulating subdomains of the third intracellular loop ( P08700 ) activated minimally when present alone but produced additive effects when present in combination . Peptides representing the amino-side P08700 and the juxtamembrane carboxyl terminus autonomously inhibited adenylate cyclase , and this response was not significantly augmented or inhibited by peptides representing other intracellular domains . Site-directed antipeptide antibodies developed against the domains of the amino terminus , first extracellular loop , amino-side P08700 , and juxtamembrane carboxyl terminus of P21554 receptors failed to influence binding of [3H]CP-55940 . However , IgG raised against the amino-side P08700 diminished the agonist-dependent inhibition of adenylate cyclase . These experiments suggest that the juxtamembrane carboxyl terminus is critical for G protein activation by P21554 cannabinoid receptors and that the amino-side P08700 also may interact with Gi proteins leading to inhibition of adenylate cyclase . Inhibition of aldehyde dehydrogenase 2 activity enhances antimycin-induced rat cardiomyocytes apoptosis through activation of MAPK signaling pathway . Aldehyde dehydrogenase 2 ( P05091 ) , a mitochondrial-specific enzyme , has been proved to be involved in oxidative stress-induced cell apoptosis , while little is known in cardiomyocytes . This study was aimed at investigating the role of P05091 in antimycin A-induced cardiomyocytes apoptosis by suppressing P05091 activity with a specific P05091 inhibitor DB02115 . Antimycin A ( 40μg/ml ) was used to induce neonatal cardiomyocytes apoptosis . DB02115 ( 60μM ) effectively inhibited P05091 activity by 50 % without own effect on cell apoptosis , and significantly enhanced antimycin A-induced cardiomyocytes apoptosis from 33.5±4.4 to 56.5±6.4 % ( Hochest method , p < 0.05 ) , and from 57.9±1.9 to 74.0±11.9 % ( FACS , p < 0.05 ) . Phosphorylation of activated MAPK signaling pathway , including extracellular signal-regulated kinase ( P27361 /2 ) , c-Jun NH2-terminal kinase ( JNK ) and p38 was also increased in antimycin A and daidzin treated cardiomyocytes compared to the cells treated with antimycin A alone . These findings indicated that modifying mitochondrial P05091 activity/expression might be a potential therapeutic option on reducing oxidative insults induced cardiomyocytes apoptosis . Tumor growth retardation and chemosensitizing action of fatty acid synthase inhibitor orlistat on T cell lymphoma : implication of reconstituted tumor microenvironment and multidrug resistance phenotype . BACKGROUND : DB01083 , a fatty acid synthase ( P49327 ) inhibitor , has been demonstrated to inhibit tumor cell survival . However , the mechanism(s) of its tumor growth retarding action against malignancies of hematological origin remains unclear . It is also not understood if the antitumor action of orlistat implicates modulated susceptibility of tumor cell to anticancer drugs . Therefore , the present investigation focuses to study the antitumor and chemosensitizing action of orlistat in a murine host bearing a progressively growing T cell lymphoma . METHODS : Tumor-bearing mice were administered with vehicle alone or containing orlistat followed by administration of PBS with or without cisplatin . Tumor progression and survival of tumor-bearing host were monitored along with analysis of tumor cell survival and apoptosis . Tumor ascitic fluid was examined for pH , NO and cytokines . Expression of genes and proteins was investigated by RT-PCR and western blot respectively . ROS was analyzed by DCFDA staining and P49327 activity by spectrophotometry . RESULTS : DB01083 administration to tumor-bearing mice resulted in tumor growth retardation , prolonged life span , declined tumor cell survival and chemosensitization to cisplatin . It was accompanied by increased osmotic fragility , modulated acidosis , expression of ROS , NO , cytokines , Q9ULC4 and VH(+) ATPase , Bcl2 , P42574 , P04637 , inhibited P49327 activity and declined expression of MDR and P21926 proteins . CONCLUSION : DB01083 manifests antitumor and chemosensitizing action implicating modulated regulation of cell survival , reconstituted-tumor microenvironment and altered MDR phenotype . GENERAL SIGNIFICANCE : These observations indicate that orlistat could be utilized as an adjunct regimen for improving antitumor efficacy of cisplatin . P04150 pathways are involved in the inhibition of astrocyte proliferation . In earlier studies , the neural cell adhesion molecule , N- P62158 , was found to inhibit the proliferation of rat astrocytes both in vitro and in vivo . To identify the gene targets involved , we used subtractive hybridization to examine changes in gene expression that occur after astrocytes are exposed to N- P62158 in vitro . While the mRNA levels for N- P62158 decreased after such treatment , the levels of mRNAs for glutamine synthetase and calreticulin increased . Since glutamine synthetase and calreticulin are known to be involved in glucocorticoid receptor pathways , we tested a number of steroids for their effects on astrocyte proliferation . Dexamethasone , corticosterone , and aldosterone were all found to inhibit rat cortical astrocyte proliferation in culture in a dose-dependent manner . DB00834 , a potent glucocorticoid antagonist , reversed the inhibitory effects of dexamethasone . These observations prompted the hypothesis that the inhibition of proliferation by N- P62158 might be mediated through the glucocorticoid receptor pathway . Consistent with this hypothesis , the inhibition of astrocyte proliferation by N- P62158 was reversed in part by a number of glucocorticoid antagonists , including DB00834 , dehydroepiandrosterone , and progesterone . In transfection experiments with cultured astrocytes , N- P62158 treatment increased the expression of a luciferase reporter gene under the control of a minimal promoter linked to a glucocorticoid response element . The enhanced activity of this construct stimulated by N- P62158 was abolished in the presence of DB00834 . The combined data suggest that astrocyte proliferation is in part regulated by alterations in glucocorticoid receptor pathways . DB00470 -induced neurotoxicity depends on P21554 receptor-mediated c-Jun N-terminal kinase activation in cultured cortical neurons . Delta9- DB00470 ( THC ) , the main psychoactive ingredient of marijuana , induces apoptosis in cultured cortical neurons . THC exerts its apoptotic effects in cortical neurons by binding to the P21554 cannabinoid receptor . The P21554 receptor has been shown to couple to the stress-activated protein kinase , c-Jun N-terminal kinase ( JNK ) . However , the involvement of specific JNK isoforms in the neurotoxic properties of THC remains to be established . The present study involved treatment of rat cultured cortical neurons with THC ( 0.005-50 microM ) , and combinations of THC with the P21554 receptor antagonist , AM 251 ( 10 microM ) and pertussis toxin ( PTX ; 200 ng ml-1 ) . Antisense oligonucleotides ( AS ) were used to deplete neurons of P45983 and P45984 in order to elucidate their respective roles in THC signalling . Here we report that THC induces the activation of JNK via the P21554 receptor and its associated G-protein , Gi/o . Treatment of cultured cortical neurons with THC resulted in a differential timeframe of activation of the P45983 and P45984 isoforms . Use of specific P45983 and P45984 AS identified activation of caspase-3 and DNA fragmentation as downstream consequences of P45983 and P45984 activation . The results from this study demonstrate that activation of the P21554 receptor induces JNK and caspase-3 activation , an increase in Bax expression and DNA fragmentation . The data demonstrate that the activation of both P45983 and P45984 isoforms is central to the THC-induced activation of the apoptotic pathway in cortical neurons . Two types of prostacyclin receptor coupling to stimulation of adenylate cyclase and phosphatidylinositol hydrolysis in a cultured mast cell line , BNu-2cl3 cells . DB01240 ( DB01240 ) -mediated signal transduction was examined in interleukin 3 ( P08700 ) -dependent BNu-2cl3 mast cells . DB01088 , a stable DB01240 analogue , induced the accumulation of intracellular DB02527 and IP3 , and an increase in the intracellular Ca2+ concentration . Pretreatment of the cells with a protein kinase C activator , 12-O-tetradecanoyl phorbol 13-acetate , suppressed the iloprost-induced IP3 accumulation and Ca2+ mobilization , but inversely potentiated the DB02527 accumulation , suggesting that neither of these signal transduction pathways of iloprosts is the result of a secondary effect of activation of the other . Removal of P08700 from the culture medium reduced the iloprost-induced IP3 accumulation and Ca2+ mobilization , while it had no effect on the iloprost-induced DB02527 accumulation at all . These results taken together suggest that BNu-2cl3 cells express two types of P43119 ; one couples to stimulation of adenylate cyclase , its expression being independent of P08700 , while the other couples to phosphatidylinositol hydrolysis , its expression being dependent on P08700 . DB01373 signaling in excystation of the early diverging eukaryote , Giardia lamblia . Excystation of Giardia lamblia , which initiates infection , is a poorly understood but dramatic differentiation induced by physiological signals from the host . Our data implicate a central role for calcium homeostasis in excystation . Agents that alter cytosolic Ca(2+) levels ( 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-tetra(acetyloxymethyl) ester , a Ca(2+) channel blocker , Ca(2+) ionophores , and thapsigargin ) strongly inhibit excystation . Treatment of Giardia with thapsigargin raised intracellular Ca(2+) levels , and peak Ca(2+) responses increased with each stage of excystation , consistent with the kinetics of inhibition . Fluorescent thapsigargin localized to a likely Ca(2+) storage compartment in cysts . The ability to sequester ions in membrane-bounded compartments is a hallmark of the eukaryotic cell . These studies support the existence of a giardial thapsigargin-sensitive Ca(2+) storage compartment resembling the sarcoplasmic/endoplasmic reticulum calcium ATPase pump-leak system and suggest that it is important in regulation of differentiation and appeared early in the evolution of eukaryotic cells . P62158 antagonists also blocked excystation . The divergent giardial calmodulin localized to the eight flagellar basal bodies/centrosomes , like protein kinase A . Inhibitor kinetics suggest that protein kinase A signaling triggers excystation , whereas calcium signaling is mainly required later , for parasite activation and emergence . Thus , the basal bodies may be a cellular control center to coordinate the resumption of motility and cytokinesis in excystation . 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 .

MH_train_1583

MH_train_1583

MH_train_1583

interacts_with DB00734?

multiple_choice

P14416 occupancy by risperidone : implications for the timing and magnitude of clinical response . The objective of the study is to investigate whether dopamine D2 receptor occupancy by risperidone and plasma levels over time can account for therapeutic efficacy and the latency period to response . Thirty-eight examinations with (123)I-IBZM single photon emission computed tomography were performed on 22 patients with schizophrenia , at diagnosis , 48 h after starting risperidone treatment and at a stable dose . DB00734 plasma levels were determined and psychopathologic evaluations ( Brief Psychiatric Rating Scale , Positive and Negative Syndrome Scale ) were carried out . No differences in the striatal/occipital ( S/O ) ratio or plasma levels were found between examinations at the 48-h time point and when a stable dose level had been established , so these parameters could not account for the latency period required for clinical response . D2 receptor occupancy at 48 h correlated positively with clinical improvement after 2 weeks of treatment . Therefore , if these results are confirmed , D2 receptor occupancy at the beginning of treatment with risperidone may be a predictor of subsequent clinical response . ( N ) -methanocarba-2MeSADP ( MRS2365 ) is a subtype-specific agonist that induces rapid desensitization of the P47900 receptor of human platelets . DB00640 diphosphate ( ADP ) initiates and maintains sustained aggregation of platelets through simultaneous activation of both the Gq-coupled P47900 receptor and the Gi-coupled Q9H244 receptor . We recently described the synthesis and P47900 receptor-specific agonist activity of ( N ) -methanocarba-2MeSADP ( MRS2365 ) . Consequences of selective activation of the P47900 receptor by MRS2365 have been further examined in human platelets . Whereas MRS2365 alone only induced shape change , addition of MRS2365 following epinephrine treatment , which activates the Gi/z-linked , alpha2A-adrenergic receptor , resulted in sustained aggregation that was indistinguishable from that observed with ADP . Conversely , the platelet shape change promoted by ADP in the presence of the P08514 /IIIa antagonist eptifibatide was similar to that promoted by MRS2365 . Preaddition of the high affinity P47900 receptor antagonist MRS2500 inhibited the effect of MRS2365 , whereas addition of MRS2500 subsequent to MRS2365 reversed the MRS2365-induced shape change . Preactivation of the P47900 receptor with MRS2365 for 2 min resulted in marked loss of capacity of ADP to induce aggregation as evidenced by a greater than 20-fold rightward shift in the concentration effect curve of ADP . This inhibitory effect of P47900 receptor activation was dependent on the concentration of MRS2365 ( EC50 = 34 nm ) . The inhibitory effect of preincubation with MRS2365 was circumvented by activation of the Gq-coupled 5- Q13049 receptor suggesting that MRS2365 induces loss of the ADP response as a consequence of desensitization of the Gq-coupled P47900 receptor . The time course of MRS2365-induced loss of aggregation response to epinephrine was similar to that observed with ADP . These results further demonstrate the P47900 receptor selectivity of MRS2365 and illustrate the occurrence of agonist-induced desensitization of the P47900 receptor of human platelets studied in the absence of Q9H244 receptor activation . 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 . In vitro effects and in vivo efficacy of a novel cyclooxygenase-2 inhibitor in cats with lipopolysaccharide-induced pyrexia . OBJECTIVE : To determine cyclooxygenase ( P36551 ) -2 selectivity , pharmacokinetic properties , and in vivo efficacy of firocoxib ( Q9NTI2 ,785,713 ) in cats . ANIMALS : 5 healthy male and 14 healthy female domestic shorthair cats . PROCEDURE : Selectivity of firocoxib for inhibiting P35354 was determined by comparing the potency for inhibiting P23219 with that of P35354 in feline blood . Pharmacokinetic properties were determined after i.v. ( 2 mg/kg ) and oral ( 3 mg/kg ) administration in male cats . In vivo efficacy was evaluated in female cats with lipopolysaccharide ( LPS ) -induced pyrexia with administration of firocoxib 1 or 14 hours before LPS challenge . RESULTS : Blood concentrations resulting in 50 % inhibition of P23219 and P35354 activity in vitro were 75 +/- 2 microM and 0.13 +/- 0.03 microM , respectively , and selectivity for inhibiting P35354 relative to P23219 was 58 . DB09217 had moderate to high oral bioavailability ( 54 % to 70 % ) , low plasma clearance ( 4.7 to 5.8 mL/min/kg ) , and an elimination half-life of 8.7 to 12.2 hours . DB09217 at doses from 0.75 to 3 mg/kg was efficacious in attenuating fever when administered to cats 1 or 14 hours before LPS challenge . CONCLUSIONS AND CLINICAL RELEVANCE : DB09217 is a potent P35354 inhibitor and is the only selective P35354 inhibitor described for use in cats to date . It is effective in attenuating febrile responses in cats when administered 14 hours before LPS challenge , suggesting it would be suitable for once-a-day dosing . Because selective P35354 inhibitors have an improved therapeutic index relative to nonselective nonsteroidal anti-inflammatory drugs in humans , firocoxib has the potential to be a safe , effective anti-inflammatory agent for cats . Monoclonal antibodies in acute lymphoblastic leukemia . With modern intensive combination polychemotherapy , the complete response ( CR ) rate in adults with acute lymphoblastic leukemia ( ALL ) is 80 % to 90 % , and the cure rate is 40 % to 50 % . Hence , there is a need to develop effective salvage therapies and combine novel agents with standard effective chemotherapy . ALL leukemic cells express several surface antigens amenable to target therapies , including P11836 , P20273 , and P15391 . Monoclonal antibodies target these leukemic surface antigens selectively and minimize off-target toxicity . When added to frontline chemotherapy , rituximab , an antibody directed against P11836 , increases cure rates of adults with Burkitt leukemia from 40 % to 80 % and those with pre-B ALL from 35 % to 50 % . Inotuzumab ozogamicin , a P20273 monoclonal antibody bound to calicheamicin , has resulted in marrow CR rates of 55 % and a median survival of 6 to 7 months when given to patients with refractory-relapsed ALL . DB09052 , a biallelic T cell engaging the CD3- P15391 monoclonal antibody , also resulted in overall response rates of 40 % to 50 % and a median survival of 6.5 months in a similar refractory-relapsed population . Other promising monoclonal antibodies targeting P11836 ( ofatumumab and obinutuzumab ) or P15391 or P11836 and bound to different cytotoxins or immunotoxins are under development . Combined modalities of chemotherapy and the novel monoclonal antibodies are under investigation . P42574 -Dependent Cleavage of the Glutamate- DB00151 Ligase Catalytic Subunit during Apoptotic Cell Death . Apoptotic cell death is usually accompanied by activation of a family of cysteine proteases termed caspases . Caspases mediate the selective proteolysis of multiple cellular targets often resulting in the disruption of survival pathways . Intracellular levels of the antioxidant glutathione ( DB00143 ) are an important determinant of cellular susceptibility to apoptosis . The rate-limiting step in DB00143 biosynthesis is mediated by glutamate-L-cysteine ligase ( GCL ) , a heterodimeric enzyme consisting of a catalytic ( P48506 ) and a modifier ( P48507 ) subunit . In this report we demonstrate that P48506 is a direct target for caspase-mediated cleavage in multiple models of apoptotic cell death . Mutational analysis revealed that caspase-mediated cleavage of P48506 occurs at DB00128 (499) within the sequence AVVD(499)G . P48506 cleavage occurs upstream of DB00151 (553) , which is thought to be important for association with P48507 . P48506 cleavage is accompanied by a rapid loss of intracellular DB00143 due to caspase-mediated extrusion of DB00143 from the cell . However , while P48506 cleavage is dependent on caspase-3 , DB00143 extrusion occurs by a caspase-3-independent mechanism . Our identification of P48506 as a target for caspase-3-dependent cleavage during apoptotic cell death suggests that this post-translational modification may represent a novel mechanism for regulating DB00143 biosynthesis during apoptosis . 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 . 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 1-beta-D-arabinofuranosylcytosine incorporation on elongation of specific DNA sequences by P06746 . DB00987 ( ara-C ) is an effective antileukemic agent which acts as an inhibitor of DNA synthesis . The precise mechanism responsible for this inhibitory effect , however , remains unclear . The present work has examined the effects of the triphosphate derivative , ara- P53007 , on purified P06746 . These studies were performed on M13 phage DNA templates of defined sequence . The results demonstrate that ara-C is incorporated into DNA by P06746 . The results also demonstrate that the incorporated ara-C residue acts as a relative chain terminator . Moreover , the relative chain terminating effects of ara-C are sequence specific . In this regard , DNA strand elongation was progressively slowed at sequences of two , three , and four contiguous sites for cytosine incorporation . We also demonstrate that the inhibitory effects of ara-C are reversed by competition with deoxycytidine-triphosphate for incorporation into the DNA strand . Taken together , these findings are consistent with structural differences of the incorporated arabinosyl moiety which alter reactivity of the 3'-terminus and thereby inhibit chain elongation . These findings also provide new insights regarding the inhibitory effects of ara-C on elongation of specific DNA sequences . Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 , CD8 and forkhead box P09131 ( Foxp3 ) , respectively . P14136 , Iba1 and P34810 -immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed . 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 . Expression of P20839 is regulated in response to mycophenolate concentration . DB04335 5'-monophosphate dehydrogenase ( IMPDH ) catalyzes de novo guanine nucleotide synthesis . DB01024 ( DB00603 ) exerts immunosuppressive effects by inhibiting IMPDH . The aim of this study was to investigate gene expressions of two IMPDH isoforms , during in vivo exposure to DB00603 . Healthy volunteers ( n=5 ) were given single doses of 100 , 250 , 500 and 1000 mg mycophenolate mofetil ( DB00688 ) . Blood was sampled pre-dose and at 1 , 2 , 4 , 6 , 8 , 12 , and 24 h post-dose . The expressions of P20839 and 2 were quantified in P01730 + cells and whole blood by real-time reverse transcription-PCR . Following DB00688 doses of 500 mg , the expression of P20839 and 2 in P01730 + cells was reduced 39 % ( P=0.043 ) and 10 % ( P=0.043 ) , respectively . Smaller reductions ( ns ) were observed after 1000 mg DB00688 . Similar trends were demonstrated for whole blood . The largest reductions of P20839 occurred at DB00603 AUC ( 0-12 h ) of 20 mg h/L . Below this , increasing DB00603 exposure correlated with larger reductions of P20839 expression ( P01730 + cells : r=-0.82 , P < 0.001 , and whole blood : r=-0.50 , P=0.04 , n=17 ) , while higher DB00603 exposure seemed to be associated with smaller reductions of expression ( P01730 + cells : r=0.42 , ns , and whole blood : r=0.77 , P=0.039 , n=8 ) . The concentration-dependent modulation of P20839 and 2 expressions by DB00603 might impact IMPDH activity . Knowledge of the regulation of the two IMPDH isoenzymes in vivo by DB00603 is of importance considering pharmacodynamic monitoring and optimization of DB00603 treatment . Methodological challenges in monitoring new treatments for rare diseases : lessons from the cryopyrin-associated periodic syndrome registry . BACKGROUND : The Q96P20 -Associated Periodic Syndromes ( CAPS ) are a group of rare hereditary autoinflammatory diseases and encompass Familial Cold Autoinflammatory Syndrome ( FCAS ) , Muckle-Wells Syndrome ( MWS ) , and Neonatal Onset Multisystem Inflammatory Disease ( NOMID ) . DB06168 is a monoclonal antibody directed against P01584 and approved for CAPS patients but requires post-approval monitoring due to low and short exposures during the licensing process . Creative approaches to observational methodology are needed , harnessing novel registry strategies to ensure Health Care Provider reporting and patient monitoring . METHODS : A web-based registry was set up to collect information on long-term safety and effectiveness of canakinumab for CAPS . RESULTS : Starting in November 2009 , this registry enrolled 241 patients in 43 centers and 13 countries by December 31 , 2012 . One-third of the enrolled population was aged < 18 ; the overall population is evenly divided by gender . Enrolment is ongoing for children . CONCLUSIONS : Innovative therapies in orphan diseases require post-approval structures to enable in depth understanding of safety and natural history of disease . The rarity and distribution of such diseases and unpredictability of treatment require innovative methods for enrolment and follow-up . Broad international practice-based recruitment and web-based data collection are practical . 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 . 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 . Evaluation of a microarray for genotyping polymorphisms related to xenobiotic metabolism and DNA repair . We present an oligonucleotide microarray ( " MetaboChip " ) based on the arrayed primer extension ( P27695 ) technique , allowing genotyping of single nucleotide polymorphisms ( SNPs ) in genes of interest for cancer susceptibility and pharmacogenetics . P27695 consists of a sequencing reaction primed by an oligonucleotide anchored with its 5' end to a glass slide and terminating one nucleotide before the polymorphic site . The extension with one fluorescently labeled dideoxynucleotide complementary to the template reveals the polymorphism . Ninety-three SNPs in 42 genes were selected among those resequenced in the context of the SNP500 project , using a set of 102 reference DNA samples from the Coriell Biorepository . Selected SNPs belong to the following genes : P00325 , P05091 , P27695 , CDKN2A , P21964 , P04798 , P05177 , Q16678 , P11509 , P33261 , P11712 , P05181 , P08684 , P14416 , P21917 , P07099 , P07992 , P18074 , Q92889 , P28715 , P30550 , O15217 , P21266 , P09211 , GSTT2 , LIG3 , Q00987 , P16455 , P05164 , NAT1 , NAT2 , P15559 , O15527 , P12004 , P06746 , Q01959 , P04179 , P04637 , P18887 , O43543 , O43542 , and O15287 . We assessed the performance of P27695 by comparing the results obtained with MetaboChip against those reported by the SNP500 . Among 88 SNPs that yielded signals , 6 showed less than 99 % of concordance , whereas 82 performed accurately , showing that P27695 is a reliable and sensitive genotyping method . 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 . Catabolic fate and pharmacokinetic characterization of trastuzumab emtansine ( DB05773 ) : an emphasis on preclinical and clinical catabolism . DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate in clinical development for the treatment of human epidermal growth factor receptor 2 ( P04626 ) -positive cancers . Herein , we describe a series of studies to assess DB05773 absorption , distribution , metabolism , and excretion ( ADME ) in rats as well as to assess human exposure to DB05773 catabolites . Following administration of unlabeled and radiolabeled DB05773 in female Sprague Dawley rats as a single dose , plasma , urine , bile and feces were assessed for mass balance , profiling and identification of catabolites . In rats , the major circulating species in plasma was DB05773 , while DM1 concentrations were low ( 1.08 to 15.6 ng/mL ) . The major catabolites found circulating in rat plasma were DM1 , [ N-maleimidomethyl ] cyclohexane-1- carboxylate-DM1 ( MCC-DM1 ) , and DB00123 -MCC-DM1 . These catabolites identified in rats were also detected in plasma samples from patients with P04626 -positive metastatic breast cancer who received single-agent DB05773 ( 3.6 mg/kg every 3 weeks ) in a phase 2 clinical study . There was no evidence of tissue accumulation in rats or catabolite accumulation in human plasma following multiple dosing . In rats , DB05773 was distributed nonspecifically to the organs without accumulation . The major pathway of DM1-containing catabolite elimination in rats was the fecal/biliary route , with up to 80 % of radioactivity recovered in the feces and 50 % in the bile . The rat DB05773 ADME profile is likely similar to the human profile , although there may be differences since trastuzumab does not bind the rat P04626 - like receptor . Further research is necessary to more fully understand the DB05773 ADME profile in humans . Neuropeptide Y selectively inhibits slow synaptic potentials in rat dorsal raphe nucleus in vitro by a presynaptic action . Neuropeptide Y ( P01303 ) has been shown to modulate synaptic transmission in both peripheral and central tissues via both pre- and postsynaptic mechanisms . In this study , we examined the effect of P01303 and its analog , peptide YY ( P10082 ) , on slow synaptic potentials in the dorsal raphe nucleus in vitro using intracellular recording and single-microelectrode voltage-clamp techniques . P01303 and P10082 inhibited both the slow P08908 receptor-mediated IPSP and the alpha 1-adrenoceptor-mediated slow EPSP while not affecting the fast , amino acid-mediated synaptic responses . P10082 also inhibited pharmacologically isolated slow synaptic responses . P01303 / P10082 appear to mediate the observed inhibitions via a presynaptic mechanism , as the postsynaptic conductances mediated by activation of P08908 receptors or alpha 1-adrenoceptors were unaffected by the peptides . P01303 / P10082 act via a different mechanism than presynaptic P28222 receptors . P01303 / P10082 probably act via presynaptic P28062 receptors , as the C-terminal fragment P01303 13-36 and the P28062 -selective agonist P06681 - P01303 are effective . Since P01303 and its receptors are present in the dorsal raphe nucleus , this peptide may act as an endogenous modulator of the state of activity of neurons in this region and may thus have a role in the modulation of neuronal output from this nucleus . 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 . 5-hydroxytryptamine 1A receptor activation protects against N-methyl-D-aspartate-induced apoptotic cell death in striatal and mesencephalic cultures . Apoptosis and glutamate-mediated excitotoxicity may play a role in the pathogenesis of many neurodegenerative disorders , including Parkinson 's disease ( PD ) . In the present study , we investigated whether stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor attenuates N-methyl-D-aspartate- ( DB01221 ) and 1-methyl-4-phenylpyridinium ( P25189 (+) ) -induced apoptotic cell death in cell culture models . A brief exposure ( 20 min ) of M213-2O striatal cells to DB01221 and glutamate produced a delayed increase in caspase-3 activity and DNA fragmentation in a dose- and time-dependent manner . DB01221 -induced caspase-3 activity and DNA fragmentation were almost completely blocked by the P08908 agonists 8-hydroxy-2-(di-n-propylamino)-tetralin ( 8-OH-DPAT ) and ( R ) -5-fluoro-8 hydroxy-2-(dipropylamino)-tetralin ( R-UH-301 ) . Additionally , the protective effects of 8-OH-DPAT and R-UH-301 on DB01221 -induced caspase-3 activation and apoptosis were reversed by pretreatment with the P08908 antagonists N- [ 2-[4-(2-methoxyphenyl)-1-piperazinyl] ethyl ] -N-(2-pyridinyl) cyclohexane carboxamide ( WAY 100635 ) and S-UH-301 , respectively . Similarly , dose- and time-dependent increases in caspase-3 activity and DNA fragmentation were observed in rat primary mesencephalic neurons after a brief exposure to DB01221 and glutamate . P42574 activation and DNA fragmentation in primary mesencephalic neurons were almost completely inhibited by 8-OH-DPAT . This neuroprotective effect of 8-OH-DPAT was reversed by WAY 100635 . Additionally , 8-OH-DPAT blocked tyrosine hydroxylase ( TH ) -positive cell death after DB01221 exposure and also almost completely attenuated the DB01221 -induced Ca(2+) influx in primary mesencephalic cultures . Furthermore , 8-OH-DPAT and R-UH-301 blocked apoptotic cell death in the primary mesencephalic neurons that were exposed to the Parkinsonian toxin P25189 (+) . Together , these results suggest that P08908 receptor stimulation may be a promising pharmacological approach in the development of neuroprotective agents for PD .

MH_train_1584

MH_train_1584

MH_train_1584

interacts_with DB01418?

multiple_choice

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 . Use of a collagen biomatrix ( TissuDura ) for dura repair : a long-term neuroradiological and neuropathological evaluation . PURPOSE : The aim of this study was to evaluate the clinical , neuroradiological , and neuropathological outcomes of patients treated with equine collagen foil ( TissuDura ) as a dura mater substitute during cranial and spinal neurosurgical procedures . MATERIALS AND METHODS : All patients treated at the Department of Neurosurgery of the Second University of Naples with TissuDura between 2005 and 2009 were included . Dural reconstruction was performed using TissuDura , overlaid 1 cm over the dural defect with additional fixation using fibrin glue . No surgical sutures were used . Patients underwent postoperative contrast-enhanced magnetic resonance scans at 1 week , 1 month , and 1 year after surgery to detect any cerebrospinal fluid ( P04141 ) leaks , infections , inflammations , or P04141 circulation in the surgical region . RESULTS : Dural reconstruction was performed in 74 patients , including 50 patients with tumors , two with P06681 neurinoma , two with acoustic neurinoma , six with Chiari I malformation , two with severe head injury , and 12 requiring spinal surgery . Clinical and neuroradiological findings were normal and no signs of graft rejection or P04141 leaks at postoperative follow-up were observed . In two cases of atypical meningioma , re-operation of the dural reconstruction was performed after 1 year . No adherences between brain and neodura were detected , and histopathological investigations demonstrated dural regeneration . CONCLUSIONS : Following dural reconstructions with TissuDura without surgical sutures , no local toxicity or complications were observed for up to 1 year . TissuDura demonstrated elasticity , non-reactivity , and good adaptability . The overlay technique using fibrin glue was simple and fast . Future studies and longer follow-up are needed to confirm the efficacy of TissuDura . Human Q14376 . Accommodation of UDP-N-acetylglucosamine within the active site . Q14376 catalyzes the interconversion of DB03501 and UDP-glucose during normal galactose metabolism . One of the key structural features in the proposed reaction mechanism for the enzyme is the rotation of a 4'-ketopyranose intermediate within the active site pocket . Recently , the three-dimensional structure of the human enzyme with bound DB00157 and UDP-glucose was determined . Unlike that observed for the protein isolated from Escherichia coli , the human enzyme can also turn over UDP-GlcNAc to UDP-GalNAc and vice versa . Here we describe the three-dimensional structure of human epimerase complexed with DB00157 and UDP-GlcNAc . To accommodate the additional N-acetyl group at the P06681 position of the sugar , the side chain of DB00174 -207 rotates toward the interior of the protein and interacts with DB00142 -199 . Strikingly , in the human enzyme , the structural equivalent of DB00135 -299 in the E. coli protein is replaced with a cysteine residue ( DB00151 -307 ) and the active site volume for the human protein is calculated to be approximately 15 % larger than that observed for the bacterial epimerase . This combination of a larger active site cavity and amino acid residue replacement most likely accounts for the inability of the E. coli enzyme to interconvert UDP-GlcNAc and UDP-GalNAc . 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 . 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 . 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 . The effect of etanercept on osteoclast precursor frequency and enhancing bone marrow oedema in patients with psoriatic arthritis . OBJECTIVE : The frequency of osteoclast precursors ( OCPF ) and the presence of bone marrow oedema ( BMO ) are potential response biomarkers in psoriatic arthritis ( PsA ) . Previous studies suggest a central role for tumour necrosis factor ( P01375 ) in the formation of osteoclast precursors . To better understand this association , the effect of etanercept on OCPF and BMO was analysed in PsA patients with erosive arthritis . METHODS : A total of 20 PsA patients with active erosive PsA were enrolled . DB00005 was administered twice weekly for 24 weeks . OCPF was measured and clinical assessments were performed at baseline , 2 , 12 and 24 weeks . Gadolinium enhanced MR images were obtained at baseline and 24 weeks . RESULTS : Significant improvements in joint score ( p < 0.001 ) , HAQ scores ( p < 0.001 ) and SF-36 parameters were observed after 6 months of therapy with etanercept compared to baseline . The median OCPF decreased from 24.5 to 9 ( p = 0.04 ) and to 7 ( p = 0.006 ) after 3 months and 6 months of treatment , respectively . MR images were available for 13 patients . The BMO volume decreased in 47 and increased in 31 sites at 6 months . No correlation was noted between OCPF , BMO and clinical parameters . CONCLUSION : The rapid decline in OCPF and overall improvement in BMO after anti-TNFalpha therapy provides one mechanism to explain the anti-erosive effects of P01375 blockade in PsA . Persistence of BMO after etanercept treatment , despite a marked clinical response , was unexpected , and suggests ongoing subchondral inflammation or altered remodelling in PsA bone . 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 . A functional candidate screen for coeliac disease genes . It is increasingly evident that different inflammatory disorders show some overlap in their pathological features , concurrence in families and individuals , and shared genetic factors . This might also be true for coeliac disease , a chronic inflammatory disorder of the gastrointestinal system , which shares two linkage regions with inflammatory bowel disease : on chromosome 5q31 ( CELIAC2 and IBD5 ) and 19p13 ( CELIAC4 and IBD6 ) . We hypothesised that these regions contain genes that contribute to susceptibility to both disorders . The overlapping 5q31 region contains only five positional candidate genes , whereas the overlapping 19p13 region has 141 genes . As the common disease gene probably plays a role in inflammation , we selected five functional candidate genes from the 19p13 region . We studied these 10 positional and functional candidate genes in our Dutch coeliac disease cohort using 44 haplotype tagging single-nucleotide polymorphisms . Two genes from 19p13 showed a small effect on familial clustering : the cytochrome P450 P13726 gene Q08477 ( P(nominal) 0.0375 , odds ratio ( OR ) 1.77 ) and P78329 ( P(nominal) 0.013 , OR 1.33 ) . Q08477 and P78329 catalyse the inactivation of leukotriene B4 ( LTB4 ) , a potent mediator of inflammation responsible for recruitment and activation of neutrophils . The genetic association of LTB4-regulating gene variants connects the innate immune response of neutrophil mobilisation with that of the established Th1 adaptive immunity present in coeliac disease patients . The findings in coeliac disease need to be replicated . Expanding genetic association studies of these cytochrome genes to other inflammatory conditions should reveal whether their causative influence extends beyond coeliac disease . 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 . P13726 -independent effects of recombinant factor VIIa on hemostasis . The molecular mechanisms responsible for the hemostatic efficacy of recombinant activated factor VII ( DB00036 ; NovoSeven , Novo Nordisk , Bagsvaerd , Denmark ) in platelet-related bleeding disorders remain unclear . The general concept is that DB00036 locally enhances thrombin generation at the site of injury , where tissue factor ( TF ) has become exposed . However , a growing amount of evidence shows that DB00036 is also able to exert its activity in a manner independent of TF . Using an in vitro flow model , we recently showed that TF-independent thrombin generation is responsible for increased platelet deposition onto injured vessels following DB00036 administration . Furthermore , it has been shown that DB00036 can restore platelet aggregation in Glanzmann 's thrombasthenia ( GT ) patients via TF-independent thrombin generation . However , the mechanism behind TF-independent thrombin generation remains to be elucidated . It is postulated that , in vivo , both the TF-dependent and TF-independent thrombin generation induced by DB00036 contribute to the control of hemorrhage in patients with platelet-related bleeding disorders and , perhaps , other causes of hemorrhagic diatheses . 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 . Immunization with inflammatory proteome of Brugia malayi adult worm induces a Th1/Th2-immune response and confers protection against the filarial infection . Mastomys coucha and jirds ( Meriones unguiculatus ) were immunized with four cytokine-stimulating SDS-PAGE resolved fractions P12259 ( 68-84 kDa ) , F6 ( 54-68 kDa ) , F10 ( 38-42 kDa ) and F14 ( 20-28 kDa ) of Brugia malayi adult worm to determine which of these fractions has the potential to influence the establishment of subsequently introduced B. malayi infection in the animals . The proteins in the fractions were analyzed by 2DE and MALDI-TOF . Immunization with F6 suppressed the establishment of third stage larva ( L(3) ) initiated infection in M. coucha ( 64 % ; P < 0.01 ) and jird ( 42 % ; P < 0.01 ) . Survival of intraperitoneally implanted adult worms in M. coucha was lowered by F6 ( 72 % ; P < 0.01 ) and F14 ( 66 % ; P < 0.05 ) but not by P12259 and F10 . Immunization with F6 intensely upregulated both Th1 ( P01579 , P01375 , P01584 , P60568 , P05231 , IgG1 , IgG2a and lymphoproliferation ) and Th2 ( IgG2b and P22301 ) responses and NO release . Immunostimulatory proteins HSP60 , intermediate filament protein , and translation elongation factor P13639 were identified in F6 fraction by 2DE and MALDI . The findings suggest that F6 protects the host from the parasite via Th1/Th2 type responses and thus holds promise for development as a vaccine . Substituted benzamides with conformationally restricted side chains . 5 . Azabicyclo[x.y.z] derivatives as Q13639 receptor agonists and gastric motility stimulants . The syntheses of benzamides containing azabicyclo[x.y.z] side chains and their Q13639 receptor agonist and 5- Q9H205 receptor antagonist properties are described . These compounds were designed to mimic higher energy conformations of quinolizidine and indolizidine . High potency was achieved for both activities although an exactly paralleling SAR was not apparent . Introduction of O and S resulted in only marginal differences in potency which was more apparent for 5- Q9H205 antagonism . The introduction of a methyl group alpha to the basic nitrogen resulted in a reduction in Q13639 receptor agonist potency . DB04917 ( 5f ) was identified for further evaluation for which both enantiomers had an identical pharmacological profile , as did an azatricyclic 9b , which contained a combination of the steric bulk of the two separate enantiomers . 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 . 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 . 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 . Structure and functions of human cerebrospinal fluid lipoproteins from individuals of different P02649 genotypes . Recent data have implicated apolipoprotein E ( apoE ) in neuritic outgrowth , synaptic stability , and Alzheimer 's disease ; these data led us to examine the normal role of apoE-containing lipoproteins in the central nervous system ( CNS ) . We isolated lipoproteins from human cerebrospinal fluid ( P04141 ) in order to examine their composition and potential functions . P04141 particles were composed of approximately one-third protein , one-third phospholipid , and one-third cholesterol . ApoE3 formed homodimers and heterodimers with apoA-II , while apoE4 , as expected , was monomeric . We addressed the function of P04141 lipoproteins with assays of cholesterol efflux and cholesterol influx . P04141 lipoproteins decreased intracellular levels of cholesterol in cholesterol-loaded fibroblasts , suggesting these particles can act to remove excess lipids from cells . P04141 lipoproteins competed for 125I-labeled LDL degradation by fibroblasts , suggesting they can also interact with the P01130 . Furthermore , P04141 lipoproteins labeled with the fluorescent dye Dil were internalized by neuroglioma cells and primary neurons and astrocytes in culture . Together , these data support a model of P04141 lipoproteins acting to remove lipids from degenerating cells and delivering lipids to cells for new membrane synthesis or storage . Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 -induced survival . This inhibition can not be overcome by increasing concentrations of P05113 and is not due to the blocking of Na+ channels by lidocaine . Here we report that one class of K+ channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 . In contrast , increasing concentrations of P08700 or granulocyte-macrophage P04141 overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 -sensitive K+ channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [3H]glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases . Effects of G- P04141 on systemic inflammation , coagulation and platelet activation in patients with acute myocardial infarction . INTRODUCTION : In the prospective , randomised , double-blind , placebo-controlled Regenerate Vital Myocardium by Vigorous Activation of Bone Marrow Stem Cells ( REVIVAL ) -2 trial patients with acute myocardial infarction ( AMI ) and successful mechanical reperfusion received granulocyte-colony stimulating factor ( DB00099 , 10 μg/kg KG s.c. ) or placebo for 5 days . Aim of this substudy was to assess the impact of G- P04141 on systemic inflammatory and procoagulant responses and platelet activation . METHODS AND RESULTS : Before and five days after DB00099 ( n=56 ) or placebo ( n=58 ) circulating cytokine concentrations of interleukin ( IL ) -1ß , P05231 , P10145 , P22301 , IL-12 and Tumor-Necrosis Factor-α ( P01375 -α were measured . P00734 fragment F1+2 and Tissue Factor activity served as a measure for activated coagulation . Platelet activation was characterized by cell surface expression of the activated fibrinogen receptor ( O95456 ) , P16109 and P29965 by flow cytometry . Administration of G- P04141 was associated with elevated P01375 -α and CRP concentrations compared to the placebo group after 5 days . Other cytokines ( IL-1ß , P05231 , P10145 , P22301 , IL-12 ) were comparable after treatment with G- P21583 or placebo . Similarly , circulating prothrombin fragments F1+2 , TF activity and platelet activation did not differ in both groups . CONCLUSION : Treatment with G- P04141 in patients with AMI was associated with enhanced proinflammatory P01375 -α and CRP levels but no activation of coagulation . DB08875 as a novel therapy for renal cell carcinoma . DB08875 / DB05153 ( Exelexis , Inc. ) has demonstrated remarkable responses in kidney cancer . Preclinical results revealed P15692 , P10721 and MET inhibition in a variety of solid tumors such as thyroid , ovarian , renal , lung , liver and prostate cancers . A phase II trial demonstrated efficacy in renal cancer with a 28 % objective response rate , stable disease rate of 62 % and median progression free survival of 14.7 months . Predominant toxicities of fatigue and diarrhea were noted . Dramatic responses in bone metastases ( three of four patients ) make the agent especially valuable for palliation in a disease , where presence of bone metastases is a predictor of worse survival . DB08875 is an emerging novel agent with promising activity in advanced kidney cancer . Randomized trials are planned in comparison with standard P15692 inhibitor therapy . Defining the role of MET overexpression would help patient selection and enrich and enhance the future evaluation of this targeted novel agent . DB08860 attenuates the PDGF-induced Q92673 /uPA receptor-mediated migration of smooth muscle cells . Statins , inhibitors of P04035 , elicit various actions on vascular cells including the modulation of proliferation and migration of smooth muscle cells ( SMCs ) . Here , we have elucidated the mechanism by which statins , in particular pitavastatin , attenuate the migration activity of SMCs . The expression of Q92673 , a member of the P01130 family and an enhancer of cell surface localization of urokinase-type plasminogen activator receptor ( Q03405 ) , is increased in cultured SMCs by treatment with DB00102 . DB08860 attenuates the DB00102 -induced surface expression of Q92673 and Q03405 . The increased migration of SMCs observed both upon overexpression of Q92673 and via stimulation of secretion of soluble Q92673 is not reversed by pitavastatin . In vivo studies showed that the SMCs expressing Q92673 in plaques are almost congruent with intimal cells expressing nonmuscle myosin heavy chain ( SMemb ) . DB08860 reduced the expression of Q92673 and SMemb , and the levels of Q92673 , Q03405 , and SMemb in cultured intimal SMCs were reduced to those seen in medial SMCs . We propose that this statin reduces PDGF-induced migration through the attenuation of the Q92673 / Q03405 system in SMCs . Modulation of the Q92673 / Q03405 system with statins suggests a novel treatment strategy for atherogenesis based on suppression of intimal SMC migration .

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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 . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . [ 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 . DB09045 for the treatment of type 2 diabetes . DB09045 is a novel glucagon-like peptide 1 ( P0C6A0 ) receptor agonist with a unique structure that supports once-weekly dosing in patients with type 2 diabetes ( T2DM ) , most of whom have a big pill burden . It appears to be efficacious in reducing hemoglobin A1c ( HbA1c ) up to 1.59 % and promotes modest weight loss up to 3 kg with a low incidence of hypoglycemia and mild to moderate gastrointestinal adverse events . Convenient weekly dosing could improve compliance and help attain sustained glycemic goals . Addressing obesity is an integral part of T2DM management and weight loss may contribute to better glycemic and cardiovascular benefits . Results of ongoing clinical trials on cardiovascular safety are important to determine the risk-to-benefit ratio . As with any drug , patient selection and ongoing monitoring will be important . If approved , dulaglutide will be one of the first weekly P43220 agonist to be available in a ready-to-use pen device with an automatic injector . 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 . Apoptosis Induction by Polygonum minus is related to antioxidant capacity , alterations in expression of apoptotic-related genes , and S-phase cell cycle arrest in HepG2 cell line . Polygonum minus ( Polygonaceae ) is a medicinal herb distributed throughout eastern Asia . The present study investigated antiproliferative effect of P. minus and its possible mechanisms . Four extracts ( petroleum ether , methanol , ethyl acetate , and water ) were prepared by cold maceration . Extracts were subjected to phytochemical screening , antioxidant , and antiproliferative assays ; the most bioactive was fractionated using vacuum liquid chromatography into seven fractions ( F1- P08709 ) . Antioxidant activity was measured via total phenolic content ( TPC ) , 2,2-diphenyl-1-picrylhydrazyl ( DPPH ) , and ferric reducing antioxidant power ( P42345 ) assays . Antiproliferative activity was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ( MTT ) assay . Most active fraction was tested for apoptosis induction and cell cycle arrest in HepG2 cells using flow cytometry and confocal microscopy . Apoptotic-related gene expression was studied by RT-PCR . Ethyl acetate extract was bioactive in initial assays . Its fraction , P08709 , exhibited highest antioxidant capacity ( TPC ; 113.16 ± 6.2 mg GAE/g extract , DPPH ; EC50 : 30.5 ± 3.2 μg/mL , P42345 ; 1169 ± 20.3 μmol Fe ( II ) /mg extract ) and selective antiproliferative effect ( IC50 : 25.75 ± 1.5 μg/mL ) . P08709 induced apoptosis in concentration- and time-dependent manner and caused cell cycle arrest at S-phase . Upregulation of proapoptotic genes ( Bax , p53 , and caspase-3 ) and downregulation of antiapoptotic gene , Bcl-2 , were observed . In conclusion , P08709 was antiproliferative to HepG2 cells by inducing apoptosis , cell cycle arrest , and via antioxidative effects . A phase II study of pralatrexate with vitamin B12 and folic acid supplementation for previously treated recurrent and/or metastatic head and neck squamous cell cancer . BACKGROUND : DB06813 ( Fotolyn(TM) ; Allos Therapeutics Inc. ) is an antifolate dihydrofolate reductase ( P00374 ) inhibitor . We conducted a phase II study of pralatrexate with folic acid and B12 supplementation in patients with recurrent and/or metastatic head and neck squamous cell cancer ( R/M HNSCC ) . PATIENTS AND METHODS : This was a single-arm , Simon optimal two stage phase II study . Patients with R/M HNSCC previously treated with chemotherapy were eligible . The study was initiated with a dosing schedule of pralatrexate 190 mg/m(2) biweekly on a 4-week cycle with vitamin supplementation . Due to toxicity concerns , the dosing was modified to 30 mg/m(2) weekly for 3 weeks in a 4-week cycle with vitamin supplementation . Radiologic imaging was to be obtained about every 2 cycles . RESULTS : Thirteen subjects were enrolled ; 12 were treated . Seven of the twelve patients had previously received ≥2 lines of chemotherapy . The most common grade 3 toxicity was mucositis ( 3 patients ) . Seven patients did not complete two cycles of therapy due to progression of disease ( 4 ) , toxicity ( 1 ) , death ( 1 ) , and withdrawal of consent ( 1 ) . Two deaths occurred : one due to disease progression and the other was an unwitnessed event that was possibly related to pralatrexate . No clinical activity was observed . The median overall survival was 3.1 months . The study was closed early due to lack of efficacy . CONCLUSIONS : DB06813 does not possess clinical activity against previously treated R/M HNSCC . Evaluation of pralatrexate in other clinical settings of HNSCC management with special considerations for drug toxicity may be warranted . 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 . Immunomodulatory drugs inhibit expression of cyclooxygenase-2 from P01375 , IL-1beta , and LPS-stimulated human PBMC in a partially P22301 -dependent manner . Immunomodulatory drugs ( IMiDs ) are potent inhibitors of P01375 and IL-1beta and elevators of P22301 production in LPS-stimulated human PBMC . They are currently in clinical trials for various diseases , including multiple myeloma , myelodysplastic syndrome , and melanoma . In the present study , we have investigated the effects of thalidomide , DB00480 and CC-4047 on the expression of P35354 by stimulated PBMC . Our results show that thalidomide and IMiDs inhibited the expression of P35354 but not the P23219 protein in LPS- P01375 and IL-1beta stimulated PBMC and shortened the half-life of P35354 mRNA in a dose-dependent manner . They also inhibited the synthesis of prostaglandin E2 from LPS-stimulated PBMC . While anti- P01375 or IL-1beta neutralizing antibodies had no effect on P35354 expression , anti- P22301 neutralizing antibody elevated the expression of P35354 mRNA , and protein from treated PBMC . These data suggest that the anti-inflammatory and anti-tumor effects of IMiDs may be due in part to elevation of P22301 production and its subsequent inhibition of P35354 expression . 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 . Comparative drug screening in Q86Y26 midline carcinoma . BACKGROUND : The Q86Y26 midline carcinoma ( NMC ) is a rare but fatal cancer for which systematic testing of therapy options has never been performed . METHODS : On the basis of disease biology , we compared the efficacy of the P50750 inhibitor flavopiridol ( FP ) with a panel of anticancer agents in NMC cell lines and mouse xenografts . RESULTS : In vitro anthracyclines , topoisomerase inhibitors , and microtubule poisons were among the most cytotoxic drug classes for NMC cells , while efficacy of the bromodomain inhibitor JQ1 varied considerably between lines carrying different O60885 ( bromodomain-containing protein 4 ) - Q86Y26 ( nuclear protein in testis ) translocations . Efficacy of FP was comparable to vincristine and doxorubicin , drugs that have been previously used in NMC patients . All three compounds showed significantly better activity than etoposide and vorinostat , agents that have also been used in NMC patients . Statins and antimetabolites demonstrated intermediate single-agent efficacy . In vivo , vincristine significantly inhibited tumour growth in two different NMC xenografts . DB03496 in vivo was significantly effective in one of the two NMC xenograft lines , demonstrating the biological heterogeneity of this disease . CONCLUSIONS : These results demonstrate that FP may be of benefit to a subset of patients with NMC , and warrant a continued emphasis on microtubule inhibitors , anthracyclines , and topoisomerase inhibitors as effective drug classes in this disease . Development of stable cell lines expressing different subtypes of GABAA receptors . The experiments reported here were motivated by our interest to express in stably-transfected cells large amounts of recombinant rat GABAA receptors . For this , we developed an original two step selection strategy , in which the first step consisted of transfecting P29320 293 cells with rat GABAA receptor alpha and beta subunits . G 418 resistant colonies isolated at this step were screened for [ 3H ] muscimol binding to select for those that coexpressed alpha- and beta-subunits . The best alpha and beta subunit expressing colony was then supertransfected with a plasmid coding for the gamma rat GABAA receptor subunit and a mutant P00374 gene . After a second round of selection , this time in presence of methotrexate , those colonies that coexpressed ternary alpha beta gamma GABAA receptor combinations were distinguished using [ 3H ] flumazenil as a probe . This strategy was applied to the isolation of 3 GABAA receptor clones , alpha 1 beta 2 gamma 2s , alpha 3 beta 2 gamma 2s and alpha 5 beta 3 gamma 2s , that expressed relatively high levels of these proteins . These 3 cell lines exhibited pharmacological and functional properties similar to cells transiently-transfected with equivalent subunit combinations . These cell lines therefore provide attractive models with which to evaluate the intrinsic activity and potency of compounds at recombinant GABAA receptor subtypes . 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 . 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 . An update on molecularly targeted therapies in second- and third-line treatment in non-small cell lung cancer : focus on P00533 inhibitors and anti-angiogenic agents . DB01248 , pemetrexed and epidermal growth factor receptor tyrosine kinase inhibitors ( gefitinib and erlotinib ) are recommended second-line therapy for advanced non-small cell lung cancer ( NSCLC ) patients with disease progression . Although erlotinib is the only recommended third-line therapy , several drugs are being used in the clinic . Recent studies have focused on combining targeted agents with approved therapies , including broad-spectrum multikinase inhibitors targeting multiple ErbB Family receptors and multitargeted anti-angiogenic agents targeting the vascular endothelial growth factor receptor , platelet-derived growth factor receptor and fibroblast growth factor receptor pathways . Here , we review targeted therapies that are being evaluated in second- and third-line settings in NSCLC , including the ErbB Family Blocker afatinib ( DB08916 ) , multityrosine kinase inhibitors ( pelitinib [ Q9Y259 -56 ] ) , neratinib [ HKI-272 ] , canertinib [ DB05424 ] , lapatinib [ GW-572016 ] , dacomitinib [ PF-299804 ] ) and multitargeted anti-angiogenic agents ( vandetanib [ DB05294 ] , sunitinib [ SU11248 ] , sorafenib [ BAY43-9006 ] , nintedanib [ BIBF1120 ] , axitinib [ AG-013736 ] , cediranib [ DB04849 ] , motesanib [ Q99217 706 ] , linifanib [ ABT869 ] and pazopanib [ DB06589 ] ) . DB01257 : A novel therapy for paroxysmal nocturnal hemoglobinuria . Paroxysmal nocturnal hemoglobinuria ( PNH ) is a rare disease caused by a somatic mutation of the P37287 gene , which results in the absence of the glycosylphosphatidylinositol-linked proteins necessary to protect cells from complement-mediated lysis . The primary clinical manifestations of PNH include intravascular hemolytic anemia , thrombosis in vessels , and bone marrow failure , which can cause pancytopenia . Treatment of PNH has been largely symptomatic until the development of eculizumab , a monoclonal antibody to the P01031 complement protein . Data from a randomized , double-blind , placebo-controlled clinical study of patients with PNH demonstrated that over a 26-week period , half of the participants receiving eculizumab experienced less hemolysis as well as stabilization of hemoglobin concentrations , and required fewer blood transfusions as compared to the placebo group . DB01257 is well tolerated , although due to its blockade of the membrane attack complex there is an increased risk of meningococcal infection . Thus , all patients must receive meningococcal vaccination prior to starting eculizumab . Clinical studies demonstrate that eculizumab may be the first successful specific therapy for the treatment of PNH . ( c ) 2007 Prous Science . All rights reserved . Influence of major structural features of tocopherols and tocotrienols on their omega-oxidation by tocopherol-omega-hydroxylase . Human cytochrome P450 4F2 ( P78329 ) catalyzes the initial omega-hydroxylation reaction in the metabolism of tocopherols and tocotrienols to carboxychromanols and is , to date , the only enzyme shown to metabolize vitamin E . The objective of this study was to characterize this activity , particularly the influence of key features of tocochromanol substrate structure . The influence of the number and positions of methyl groups on the chromanol ring , and of stereochemistry and saturation of the side chain , were explored using HepG2 cultures and microsomal reaction systems . Human liver microsomes and microsomes selectively expressing recombinant human P78329 exhibited substrate activity patterns similar to those of HepG2 cells . Although activity was strongly associated with substrate accumulation by cells or microsomes , substantial differences in specific activities between substrates remained under conditions of similar microsomal membrane substrate concentration . Methylation at P01031 of the chromanol ring was associated with markedly low activity . Tocotrienols exhibited much higher Vmax values than their tocopherol counterparts . Side chain stereochemistry had no effect on omega-hydroxylation of DB00163 ( alpha-TOH ) by any system . Kinetic analysis of microsomal P78329 activity revealed Michaelis-Menten kinetics for alpha-TOH but allosteric cooperativity for other vitamers , especially tocotrienols . Additionally , alpha-TOH was a positive effector of omega-hydroxylation of other vitamers . These results indicate that P78329 -mediated tocopherol-omega-hydroxylation is a central feature underlying the different biological half-lives , and therefore biopotencies , of the tocopherols and tocotrienols . 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 . Ribavirin-induced intracellular GTP depletion activates transcription elongation in coagulation factor VII gene expression . Coagulation FVII ( Factor VII ) is a vitamin K-dependent glycoprotein synthesized in hepatocytes . It was reported previously that FVII gene ( P08709 ) expression was up-regulated by ribavirin treatment in hepatitis C virus-infected haemophilia patients ; however , its precise mechanism is still unknown . In the present study , we investigated the molecular mechanism of ribavirin-induced up-regulation of P08709 expression in HepG2 ( human hepatoma cell line ) . We found that intracellular GTP depletion by ribavirin as well as other IMPDH ( inosine-5'-monophosphate dehydrogenase ) inhibitors , such as mycophenolic acid and 6-mercaptopurine , up-regulated P08709 expression . FVII mRNA transcription was mainly enhanced by accelerated transcription elongation , which was mediated by the P-TEFb ( positive-transcription elongation factor b ) complex , rather than by promoter activation . Ribavirin unregulated P55199 ( eleven-nineteen lysine-rich leukaemia ) 3 mRNA expression before P08709 up-regulation . We observed that ribavirin enhanced Q9HB65 recruitment to P08709 , whereas knockdown of Q9HB65 diminished ribavirin-induced FVII mRNA up-regulation . Ribavirin also enhanced recruitment of P50750 ( cyclin-dependent kinase 9 ) and Q9UHB7 to P08709 . These data suggest that ribavirin-induced intracellular GTP depletion recruits a super elongation complex containing P-TEFb , Q9UHB7 and Q9HB65 , to P08709 , and modulates FVII mRNA transcription elongation . Collectively , we have elucidated a basal mechanism for ribavirin-induced FVII mRNA up-regulation by acceleration of transcription elongation , which may be crucial in understanding its pleiotropic functions in vivo . 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 . Gender influences the class III and V β-tubulin ability to predict poor outcome in colorectal cancer . PURPOSE : Colorectal cancer is one of the deadliest diseases in Western countries . To predict the outcome of therapy , we assessed the role of class III ( Q13509 ) and class V β-tubulin ( Q9BUF5 ) as predictive biomarkers . EXPERIMENTAL DESIGN : Using immunohistochemistry and nanofluidics , the expression of Q13509 and Q9BUF5 was assessed in two cohorts of 180 and 134 patients , respectively . The P05093 RS743572 was genotyped to identify GG carriers with enhanced androgen levels . Q13509 and Q9BUF5 were investigated in 22 colorectal cancer cell lines in basal conditions and after serum starvation , the latter serving as activator of this prosurvival pathway . To ascertain the role of androgen receptor ( AR ) in such regulation , we silenced AR and checked Q13509 and Q9BUF5 expression and sensitivity to chemotherapy . RESULTS : There was a link between poor survival , the expression of Q13509 / Q9BUF5 , and AR only in females . Conversely , only in males carriers of the GG phenotype exhibited the worst outcome . Importantly , male cell lines were resistant to serum starvation and exhibited higher levels of Q9BUF5 , thereby suggesting that the pathway is activated by androgens . In female cells this phenomenon was absent . In both genders , AR was the main driver of Q13509 / Q9BUF5 expression , as constitutive silencing of AR was associated with downregulation of Q13509 / Q9BUF5 expression and increased sensitivity to oxaliplatin and SN-38 . CONCLUSIONS : The involvement of androgens in the Q13509 pathway opens the way for clinical trials to assess the efficacy of antiandrogens for increasing the efficacy of chemotherapy in male colorectal cancer patients . 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 .

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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 . Polymorphisms in one-carbon metabolism and trans-sulfuration pathway genes and susceptibility to bladder cancer . We have previously reported significant inverse associations between bladder cancer risk and dietary intake of vitamins B2 , B6 , B12 , folate and protein in a hospital-based bladder cancer case-control study conducted in Spain ( 1,150 cases;1,149 controls ) . Because these dietary factors are involved in the one-carbon metabolism pathway , we evaluated associations between bladder cancer risk and 33 single nucleotide polymorphisms ( SNPs ) in 8 genes ( P35520 , CTH , P42898 , Q99707 , Q9UBK8 , P34896 , P41440 and P04818 ) and interactions with dietary variables involved in this pathway . Two SNPs in the CTH gene were significantly associated with bladder cancer risk . OR ( 95 % CI ) for heterozygous and the homozygous variants compared to homozygous wild-type individuals were : 1.37 ( 1.04-1.80 ) IVS3-66 A > C and 1.22 ( 1.02-1.45 ) IVS10-430 C > T . Because the CTH gene is important for glutathione synthesis , we examined interactions with the P09488 gene , which codes for glutathione S-transferase muu . Increased risk for individuals with the IVS10-430 CT or TT genotype was limited to those with the P09488 null genotype ( p-interaction = 0.02 ) . No other SNPs were associated with risk of bladder cancer . These findings suggest that common genetic variants in the one-carbon pathway may not play an important role in the etiology of bladder cancer . However , our results provide some evidence that variation in glutathione synthesis may contribute to risk , particularly among individuals who carry a deletion in P09488 . Additional work is needed to comprehensively evaluate genomic variation in CTH and related genes in the trans-sulfuration pathway and bladder cancer risk . 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 . 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 . 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 . 2(3H)-benzoxazolone and bioisosters as " privileged scaffold " in the design of pharmacological probes . The 2(3H)-benzoxazolone heterocycle and its bioisosteric surrogates ( such as 2(3H)-benzothiazolinone , benzoxazinone , etc. ) have received considerable attention from the medicinal chemists owing to their capacity to mimic a phenol or a catechol moiety in a metabolically stable template . These heterocycles and pyrocatechol have indeed similar pKa 's , electronic charge distribution , and chemical reactivity . Therapeutic applications of this template are very broad , and range from analgesic anti-inflammatory compounds ( including P37231 antagonists ) to antipsychotic and neuroprotective anticonvulsant compounds . High affinity ligands have been obtained also for dopaminergic ( D2 and D4 ) , serotoninergic ( P08908 and P28223 ) , sigma-1 and sigma-2 receptors . Owing to the high number of positive hits encountered with this heterocycle and its congeners , 2(3H)-benzoxazolone template certainly deserves the title of " privileged scaffold " in medicinal chemistry . 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 . P35354 inhibitors ameliorate experimental autoimmune encephalomyelitis through modulating P01579 and P22301 production by inhibiting T-bet expression . The P35354 inhibitors DB00533 ( Rof ) and DB01283 ( Lum ) were evaluated in experimental autoimmune encephalomyelitis ( EAE ) , the model of multiple sclerosis ( MS ) . Administration of Rof and Lum significantly reduced the incidence and severity of EAE , which was associated with the inhibition of Q16653 35-55 lymphocyte recall response , anti- Q16653 35-55 T cell responses , and modulation of cytokines production . In vitro Rof and Lum inhibited primary T cells proliferation and modulated cytokine production . These findings highlight the fact that Rof and Lum likely prevents EAE by modulating Th1/Th2 response , and suggest its utility in the treatment of MS and other autoimmune diseases . The ability to accumulate deoxyuridine triphosphate and cellular response to thymidylate synthase ( TS ) inhibition . P04818 ( TS ) is an important enzyme catalysing the reductive methylation of DB03800 to dTMP that is further metabolized to dTTP for DNA synthesis . Loss of viability following TS inhibition occurs as a consequence of depleted dTTP pools and at least in some cell lines , accumulation of dUTP and subsequent misincorporation of uracil into DNA . The expansion in dUTP pools is largely determined by the expression of the pyrophosphatase , P33316 . Our previous work has shown that following TS inhibition the ability to accumulate dUTP was associated with an earlier growth inhibitory effect . 3 human lung tumour cell lines and HT29 human colon tumour cells transfected with P33316 have been used to investigate the relationship between loss of viability following TS inhibition and dUTP accumulation . Cell cycle arrest typical of TS inhibition was an early event in all cell lines and occurred irrespective of the ability to accumulate dUTP or p53 function . However , a large expansion of dUTP pools was associated with mature DNA damage ( 4 h ) and an earlier loss of viability following TS inhibition compared to cells in which dUTP pools were not expanded . In A549 cells damage to mature DNA may have been exacerbated by significantly higher activity of the excision repair enzyme , uracil-DNA glycosylase . Consistent with results using different inhibitors of TS , transfection of P33316 into HT29 cells significantly reduced the cytotoxicity of a 24 h but not 48 h exposure to ZD9331 . Although loss of viability can be mediated through dTTP deprivation alone , the uracil misincorporation pathway resulted in an earlier commitment to cell death . The relevance of this latter pathway in the clinical response to TS inhibitors deserves further investigation . Effects of short- and long-term risperidone treatment on prolactin levels in children with autism . BACKGROUND : The effects of short- and long-term risperidone treatment on serum prolactin were assessed in children and adolescents with autism . METHODS : Patients with autism ( N = 101 , 5-17 years of age ) were randomized to an 8-week trial of risperidone or placebo and 63 then took part in a 4-month open-label follow-up phase . Serum samples were obtained at Baseline and Week-8 ( N = 78 ) , and at 6-month ( N = 43 ) and 22-month ( N = 30 ) follow-up . Serum prolactin was determined by immunoradiometric assay ; dopamine type-2 receptor ( P14416 ) polymorphisms were genotyped . RESULTS : Baseline prolactin levels were similar in the risperidone ( N = 42 ) and placebo ( N = 36 ) groups ( 9.3 +/- 7.5 and 9.3 +/- 7.6 ng/ml , respectively ) . After 8 weeks of risperidone , prolactin increased to 39.0 +/- 19.2 ng/ml , compared with 10.1 +/- 8.8 ng/ml for placebo ( p < .0001 ) . P01236 levels were also significantly increased at 6 months ( 32.4 +/- 17.8 ng/ml ; N = 43 , p < .0001 ) and at 22 months ( N = 30 , 25.3 +/- 15.6 ng/ml , p < .0001 ) . P01236 levels were not associated with adverse effects and P14416 alleles ( Taq1A , -141C Ins/Del , C957T ) did not significantly influence baseline levels or risperidone-induced increases in prolactin . CONCLUSIONS : DB00734 treatment was associated with two- to four-fold mean increases in serum prolactin in children with autism . Although risperidone-induced increases tended to diminish with time , further research on the consequences of long-term prolactin elevations in children and adolescents is needed . 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 . DB09074 , P09874 inhibitor in ovarian cancer . INTRODUCTION : Ovarian cancer is the most important cause of gynecological cancer-related mortality . Conventional treatments for advanced or recurrent disease offer limited results in terms of long-term responses and survival . Researches have recently focused on target therapies , which represent a new , promising , therapeutic approach , able to maximizing tumor kill and minimizing toxicity . The family of polyadenosine diphosphate-ribose polymerase ( PARP ) inhibitors is currently one of the most hopeful and investigated alternatives . AREAS COVERED : Preclinical and clinical studies of DB09074 , the most investigated PARP inhibitor in ovarian cancer , are analyzed and discussed . Data were obtained by searching for all English peer-reviewed articles on Medline , on Cochrane Database and all on-going Phase I and II studies registered on National Cancer Institute Clinical Trials ; also any related abstracts recently presented on DB09074 at major international congresses will be included . EXPERT OPINION : Bad prognosis and drug resistance usually affect ovarian cancer . Recent trends toward the knowledge of molecular-specific pathways have produced new target drugs . PARP inhibition mediated by DB09074 in P38398 ( breast cancer 1 ) and P51587 ( breast cancer 2 ) -mutated and in sporadic ovarian cancer represents a promising field of investigation . Further studies are needed to confirm initial exciting results . 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 . P43490 regulates cell survival through autophagy in cardiomyocytes . DB02701 adenine dinucleotide ( NAD(+) ) acts as a transfer molecule for electrons , thereby acting as a key cofactor for energy production . NAD(+) also serves as a substrate for cellular enzymes , including poly ( ADPribose ) polymerase ( PARP ) -1 and Sirt1 . Activation of P09874 by DNA damage depletes the cellular pool of NAD(+) , leading to necrotic cell death . NAD(+) in the nucleus enhances the activity of Sirt1 , thereby modulating transcription . NAD(+) is either synthesized de novo from amino acids , namely tryptophan and aspartic acid , or resynthesized from NAD(+) metabolites , such as nicotinamide ( NAM ) , through the salvage pathway . NAM phosphoribosyltransferase ( P43490 ) is a rate-limiting enzyme in the NAD(+) salvage pathway . We have recently demonstrated that P43490 is an important regulator of NAD(+) and autophagy in cardiomyocytes . Here we discuss the role of P43490 in regulating autophagy and potential mechanisms by which NAD(+) regulates autophagy in the heart . Functional significance of subtypes of 5-HT receptors in the rat spinal reflex pathway . 1. The functional significance of subtypes of 5-hydroxytryptamine ( 5-HT ) receptors was studied in the rat spinal reflex pathway . 2 . Ketanserin had no effect on the mono- ( Q9UBK8 ) or polysynaptic reflex ( PSR ) in spinal rats , but decreased the PSR in intact rats . 3 . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and 5-methoxy-N,N-dimethyltryptamine ( 5-MeODMT ) decreased the Q9UBK8 and increased the PSR in spinal rats . 4 . Ketanserin antagonized the effects of 5-MeODMT without antagonizing the effects of 8-OH-DPAT . 5 . Cinanserin had similar effects to those of ketanserin . 6 . These results suggest that both P08908 and 5-HT2 receptors mediate Q9UBK8 inhibition and PSR augmentation in the spinal reflexes of spinal rats , and that the 5-HT2 receptor has a supraspinal tonic excitatory influence on the PSR in intact rats . Effect of tumour necrosis factor-alpha on estrogen metabolic pathways in breast cancer cells . P01375 -alpha ( P01375 -α ) is a proinflammatory cytokine that has been linked to breast cancer development . Estrogen metabolic pathway is also involved in breast carcinogenesis and DNA adducts formation . In this study we investigated the effect of P01375 -α on the estrogen metabolic pathway in MCF-7 , a breast cancer cell line . Capillary liquid chromatography/mass spectrometry ( LC/MS ) and High performance liquid chromatography ( HPLC ) were used for analysis of estrogen metabolites and estrogen-DNA adducts levels respectively . Reporter gene assay , Real time reverse transcription polymerase chain reaction ( real time RT-PCR ) and Western blot were used to assess the expression of estrogen metabolizing genes and enzymes . P01375 -α significantly increased the total EM and decreased the estrone ( E1 ) / 17-β estradiol ( E2 ) ratio . Moreover , it altered the expression of genes and enzymes involved in E2 activation and deactivation pathways e.g. Cytochrome P-450 1A1 ( P04798 ) , Cytochrome P-450 1B1 ( Q16678 ) , Catechol-O-methyl transferase ( P21964 ) and DB02701 adenine dinucleotide phosphate-quinone oxidoreductase 1 ( P15559 ) . In addition , there were increased levels of some catechol estrogens e.g. 4-hydroxy-estrone ( 4-OHE1 ) and 2-hydroxyestradiol ( 2-OHE2 ) with decreased levels of methylated catechols e.g. 2-methoxy estradiol ( DB02342 ) . DNA adducts especially 4-OHE1-[2]-1-N3 DB00173 was significantly increased . P01375 -α directs the estrogen metabolism into more hormonally active and carcinogenic products in MCF-7 . This may implicate a new possible explanation for inflammation associated breast cancer . Effects of pituitary hormones on the prostate . BACKGROUND : Although essential , androgens alone are not sufficient to induce normal growth and functionality of the prostate . Nonandrogenic hormones must also be involved in the proliferation of the prostate cancer cells which do not respond to antiandrogenic therapy and which thus become androgen-independent . P01236 , but also growth hormone and luteinizing hormone , are potentially able to act on both normal and abnormal prostatic cells . METHODS : In this review we summarize data from the literature concerning the physiological and pathological implications of prolactin , growth hormone , and luteinizing hormone on the prostate . RESULTS : In rodent prostates , prolactin and growth hormone can induce a variety of effects independently of androgens ( e.g. , transactivation of certain genes , or synthesis of the major secretion products ) . Moreover , hyperprolactinemia is responsible for inflammation and dysplasia of the gland , while growth hormone promotes the development of prostate tumors in vivo in the mouse and rat . Growth hormone acts on the gland directly , through prostatic growth hormone receptors , and/or indirectly via the stimulation of insulin-like growth factor-I ( P05019 ) synthesis in the liver . P22888 is expressed in rat and human prostates . DB00044 increases the amount of various transcripts in the rat prostate through an androgen-independent pathway . CONCLUSIONS : P01236 , growth hormone , and luteinizing hormone , alone or synergistically with androgens , play physiologically significant roles in the normal prostate . The involvement of these hormones in the development of benign prostatic hyperplasia and prostatic carcinoma is an issue that needs to be addressed . The spinal 5-HT system contributes to the generation of fictive locomotion in lamprey . Activation of DB01221 receptors evokes sustained fictive locomotion in the isolated spinal cord of the sea lamprey Petromyzon marinus ( P. marinus ) , but in the river lamprey Lampetra fluviatilis ( L. fluviatilis ) the ventral root activity is often irregular . A previous study showed that the number of 5-HT immunoreactive fibres , neurones and varicosities are much lower in the spinal cord of L. fluviatilis than in P. marinus . To further analyse the underlying mechanisms , the present study investigated the role of the 5-HT system in stabilising fictive locomotion . In P. marinus a blockade of P08908 receptors by spiperone reversibly increased the frequency and the coefficient of variation . This implies that there is an endogenous release of 5-HT during fictive locomotion that is important for the generation of locomotor activity . In L. fluviatilis bath applied DB01221 or D-glutamate evoked in most cases irregular activity . An addition of 5-HT ( 0.5-2 microM ) rapidly stabilised the burst generation and led to a sustained fictive locomotion . In a split-bath configuration , DB01221 administered to the rostral part of the spinal cord in P. marinus evoked fictive locomotion in both the rostral part and the first few segments of the caudal part . When spiperone was added to the caudal part , the burst activity changed into tonic activity within 10 min . Taken together , these results indicate that activity in the intrinsic 5-HT system in the lamprey spinal locomotor network contributes significantly to the rhythm generation . The quantitative differences with regard to the 5-HT plexus between P. marinus and L. fluviatilis may account for the observed discrepancy between the two species .

MH_train_1587

MH_train_1587

MH_train_1587

interacts_with DB00007?

multiple_choice

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 . The migration of luteinizing hormone-releasing hormone neurons in the developing rat is associated with a transient , caudal projection of the vomeronasal nerve . DB00044 -releasing hormone ( P01148 ) neurons originate in the olfactory placode and vomeronasal organ and migrate to the brain from embryonic day 14 ( Q14207 ) in the rat . We investigated the development of the vomeronasal nerve and its role as a guide for migrating P01148 neurons . Using fluorescent , lipophilic dye tracing methods , we observed axons that emerge from the vomeronasal organ and cross the nasal septum as several large fascicles . At Q14207 -15 , these fascicles converge as they enter the region of the cribriform plate and subsequently disperse , projecting dorsally and caudally across the olfactory bulb and rostral forebrain . At E16 , the dorsal branch of the vomeronasal nerve forms a more tightly fasciculated projection ; the caudal fibers remain dispersed , extending along the medial forebrain . The number of caudally directed axons decreases during development , leaving four or five present at postnatal day 4 ( P4 ) . Immunohistochemical studies indicate that the vomeronasal nerve can be divided into four spatially distinct subpopulations of fibers . One subset , composed of caudal fibers that terminate in the lamina terminalis , selectively expresses TAG-1 , a transient axonal surface glycoprotein and PSA-N- P62158 , a highly polysialated form of neural cell adhesion molecule . The extension and subsequent retraction of this branch of the vomeronasal nerve corresponds spatially and temporally with the migration of P01148 neurons from the nasal cavity to the brain . Our studies show that between Q14207 and E18 , P01148 neurons migrate in contact with the TAG-1+ , PSA-N- P62158 + caudal branch of the vomeronasal nerve . 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 . 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 . P05019 induces the uncoupling protein gene expression in fetal rat brown adipocyte primary cultures : role of C/EBP transcription factors . Fetal rat brown adipocytes proliferate and reach confluence when cultured in the presence of serum , almost losing their expression of the tissue-specific gene uncoupling protein ( P25874 ) . Confluent cells in a serum-free medium induced the expression of P25874 in response to 1.4 nM P05019 at 72 h , in a time-dependent manner . This effect was not produced by 1 nM insulin . However , insulin but not P05019 induced the expression of the lipogenic marker malic enzyme , suggesting that P05019 but not insulin is involved in the thermogenic differentiation process of fetal brown adipocytes . Furthermore , the expression of C/EBP transcription factors was present in fetal brown adipocytes prior culture , decreasing during cell proliferation throughout culture . After confluence , insulin induced the expression of P49715 and delta , whereas the expression of P17676 remained essentially unmodified . P05019 induced P49715 but decreased P49716 , this might indicate that both transcription factors play a reciprocal role in the expression of the tissue specific P25874 gene . 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 . Q13507 -like protein and vitamin D receptor mediate DB00136 -induced Q5T124 influx in muscle cells . 1alpha,25-Dihydroxy-Vitamin-D3 ( 1alpha,25(OH)2- DB00169 ) stimulates in skeletal muscle cells Ca2+ release from inner stores and influx through both voltage-dependent and store-operated Ca2+ ( Q5T124 , CCE ) channels . We investigated the involvement of TRPC proteins and Vitamin D receptor ( P11473 ) in CCE induced by DB00136 in chick muscle cells . Two fragments were amplified by RT-PCR , exhibiting approximately 80 % sequence homology with mammalian Q13507 /6/7 . Northern and Western blots employing a Q13507 -probe and anti- Q13507 antibodies , respectively , confirmed endogenous expression of a Q13507 -like protein of 140 kDa . Spectrofluorimetric measurements in Fura-2 loaded cells showed reduced CCE and Mn2+ entry in response to either thapsigargin or DB00136 upon transfection with anti- Q13507 /6/7 antisense oligodeoxynucleotides ( ODNs ) . Transfection with anti- P11473 antisense ODNs diminished DB00136 -dependent Ca2+ and Mn2+ influx . Co-immunoprecipitation of Q13507 -like protein and P11473 under non-denaturating conditions was observed . We propose that endogenous Q13507 -like proteins and the P11473 participate in the modulation of CCE by DB00136 in muscle cells , which could be mediated by an interaction between these proteins . Lobe-specific calcium binding in calmodulin regulates endothelial nitric oxide synthase activation . BACKGROUND : Human endothelial nitric oxide synthase ( P29474 ) requires calcium-bound calmodulin ( P62158 ) for electron transfer but the detailed mechanism remains unclear . METHODOLOGY/PRINCIPAL FINDINGS : Using a series of P62158 mutants with E to Q substitution at the four calcium-binding sites , we found that single mutation at any calcium-binding site ( B1Q , B2Q , B3Q and B4Q ) resulted in ∼2-3 fold increase in the P62158 concentration necessary for half-maximal activation ( EC50 ) of citrulline formation , indicating that each calcium-binding site of P62158 contributed to the association between P62158 and P29474 . DB00155 formation and cytochrome c reduction assays revealed that in comparison with P29475 or P35228 , P29474 was less stringent in the requirement of calcium binding to each of four calcium-binding sites . However , lobe-specific disruption with double mutations in calcium-binding sites either at N- ( B12Q ) or at C-terminal ( B34Q ) lobes greatly diminished both P29474 oxygenase and reductase activities . Gel mobility shift assay and flavin fluorescence measurement indicated that N- and C-lobes of P62158 played distinct roles in regulating P29474 catalysis ; the C-terminal EF-hands in its calcium-bound form was responsible for the binding of canonical P62158 -binding domain , while N-terminal EF-hands in its calcium-bound form controlled the movement of Q68DA7 domain . Limited proteolysis studies further demonstrated that B12Q and B34Q induced different conformational change in P29474 . CONCLUSIONS : Our results clearly demonstrate that P62158 controls P29474 electron transfer primarily through its lobe-specific calcium binding . Prevention of alcohol-induced learning deficits in fetal alcohol syndrome mediated through DB01221 and GABA receptors . OBJECTIVE : Vasoactive intestinal peptide ( P01282 ) -related peptides prevented the learning deficit in the offspring in a model for fetal alcohol syndrome . We evaluated whether the mechanism of the peptide protection included Q13224 , Q12879 , and GABAAalpha5 . STUDY DESIGN : Timed , pregnant C57BL6/J mice were injected on gestational day 8 with alcohol ( 0.03 mL/kg ) , placebo , or alcohol plus peptides . Embryos were harvested after 6 hours , 24 hours , and on gestational day 18 . Some of the litters were allowed to deliver , and the adult brains harvested after the offspring were tested for learning . Calibrator-normalized relative real-time polymerase chain reaction ( PCR ) was performed using primers for Q13224 , Q12879 , and GABAAalpha5 with P04406 standardization . Statistic : analysis of variance ( Q9UNW9 ) and Fisher PLSD , P < .05 was considered significant . RESULTS : In the embryo , the peptides prevented Q13224 rise ( P < .001 ) at 6 hours , Q13224 down-regulation ( P = .002 ) , and GABAAalpha5 decrease ( P < .01 ) on gestational day 18 . In the adult , the peptides prevented Q13224 down-regulation ( P = .01 ) and Q12879 up-regulation ( P < .001 ) . CONCLUSION : P01282 -related peptides prevented alcohol-induced changes in Q13224 , Q12879 , and GABAAalpha5 . This may explain , at least in part , the peptides ' prevention of alcohol-induced learning deficits . Transcriptional activation of human mu-opioid receptor gene by insulin-like growth factor-I in neuronal cells is modulated by the transcription factor Q13127 . The human mu-opioid receptor gene ( P35372 ) promoter contains a DNA sequence binding the repressor element 1 silencing transcription factor ( Q13127 ) that is implicated in transcriptional repression . We investigated whether insulin-like growth factor I ( P05019 ) , which affects various aspects of neuronal induction and maturation , regulates P35372 transcription in neuronal cells in the context of the potential influence of Q13127 . A series of P35372 -luciferase promoter/reporter constructs were transfected into two neuronal cell models , neuroblastoma-derived SH-SY5Y cells and PC12 cells . In the former , endogenous levels of human mu-opioid receptor ( hMOPr ) mRNA were evaluated by real-time PCR . P05019 up-regulated P35372 transcription in : PC12 cells lacking Q13127 , in SH-SY5Y cells transfected with constructs deficient in the Q13127 DNA binding element , or when Q13127 was down-regulated in retinoic acid-differentiated cells . P05019 activates the signal transducer and activator of transcription-3 signaling pathway and this transcription factor , binding to the signal transducer and activator of transcription-1/3 DNA element located in the promoter , increases P35372 transcription . We propose that a reduction in Q13127 is a critical switch enabling P05019 to up-regulate hMOPr . These findings help clarify how hMOPr expression is regulated in neuronal cells . 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 . Evidence for a link between histone deacetylation and Ca²+ homoeostasis in sphingosine-1-phosphate lyase-deficient fibroblasts . Embryonic fibroblasts from Q14703 ( sphingosine-1-phosphate ) lyase-deficient mice [ Sgpl1-/- MEFs ( mouse embryonic fibroblasts ) ] are characterized by intracellular accumulation of Q14703 , elevated cytosolic [Ca2+]i and enhanced Ca2+ storage . Since Q14703 , produced by sphingosine kinase 2 in the nucleus of MCF-7 cells , inhibited HDACs ( histone deacetylases ) [ Hait , Allegood , Maceyka , Strub , Harikumar , Singh , Luo , Marmorstein , Kordula , Milstein et al. ( 2009 ) Science 325 , 1254-1257 ] , in the present study we analysed whether Q14703 accumulated in the nuclei of Q14703 lyase-deficient MEFs and caused HDAC inhibition . Interestingly , nuclear concentrations of Q14703 were disproportionally elevated in Sgpl1-/- MEFs . HDAC activity was reduced , acetylation of histone 3-Lys9 was increased and the HDAC-regulated gene P38936 cyclin-dependent kinase inhibitor was up-regulated in these cells . Furthermore , the expression of Q13547 and O15379 was reduced in Sgpl1-/- MEFs . In wild-type MEFs , acetylation of histone 3-Lys9 was increased by the Q14703 lyase inhibitor 4-deoxypyridoxine . The non-specific HDAC inhibitor trichostatin A elevated basal [Ca2+]i and enhanced Ca2+ storage , whereas the Q13547 /2/3 inhibitor DB05651 elevated basal [Ca2+]i without influence on Ca2+ storage in wild-type MEFs . Overexpression of Q13547 or Q92769 reduced the elevated basal [Ca2+]i in Sgpl1-/- MEFs . Taken together , Q14703 lyase-deficiency was associated with elevated nuclear Q14703 levels , reduced HDAC activity and down-regulation of HDAC isoenzymes . The decreased HDAC activity in turn contributed to the dysregulation of Ca2+ homoeostasis , particularly to the elevated basal [Ca2+]i , in Sgpl1-/- MEFs . DB06626 for renal cell carcinoma . BACKGROUND : The approval of sunitinib , sorafenib and temsirolimus has dramatically altered the management of renal cell carcinoma ( RCC ) . DB00112 plus IFN may also be added to the therapeutic armamentarium . DB06626 ( AG-013736 ) is an oral and selective tyrosine kinase inhibitor . OBJECTIVE : Data supporting the development of axitinib for RCC are reviewed . METHODS : Preclinical and clinical data available for axitinib for RCC are presented . RESULTS : DB06626 inhibits P17948 , P35968 and P35916 with picomolar potencies , and P16234 , P09619 and c-kit with nanomolar potencies . Phase II clinical trials of axitinib in pretreated RCC following sorafenib or cytokine treatment have demonstrated promising activity accompanied by a favorable toxicity profile . Further development of axitinib for RCC is warranted . N-methyl-D-aspartate receptor antagonists enhance histamine neuron activity in rodent brain . The modulation of histamine neuron activity by various non-competitive DB01221 -receptor antagonists was evaluated by changes in tele-methylhistamine ( t-MeHA ) levels and histidine decarboxylase ( hdc ) mRNA expression induced in rodent brain . The DB01221 open-channel blockers phencyclidine ( PCP ) and MK-801 enhanced t-MeHA levels in mouse brain by 50-60 % . DB08954 , which interacts with polyamine sites of Q13224 -containing DB01221 receptors , had no effect . PCP also increased hdc mRNA expression in the rat tuberomammillary nucleus . The enhancement of t-MeHA levels elicited by MK-801 ( ED50 of approximately 0.1 mg/kg ) was observed in the hypothalamus , cerebral cortex , striatum and hippocampus . Control t-MeHA levels and the t-MeHA response to MK-801 were not different in male and female mice . Double immunostaining for HDC and DB01221 receptor subunits showed that histamine neurons of the rat tuberomammillary nucleus express DB01221 receptor subunit 1 ( Q9UHB4 ) with DB01221 receptor subunit 2A ( Q12879 ) and DB01221 receptor 2B subunit ( Q13224 ) . In addition , immunoreactivity for the neuronal glutamate transporter P43005 was observed near most histaminergic perikarya . Hence , these findings support the existence of histamine/glutamate functional interactions in the brain . The increase in histamine neuron activity induced by DB01221 receptor antagonists further suggests a role of histamine neurons in psychotic disorders . In addition , the decrease in MK-801-induced hyperlocomotion observed in mice after administration of ciproxifan further strengthens the potential interest of H3-receptor antagonist/inverse agonists for the symptomatic treatment of schizophrenia . 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 . 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 . 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 . Enhanced vasoactive intestinal peptide-induced prolactin secretion from anterior pituitary cells of incubating turkeys ( Meleagris gallopavo ) . During incubation , female turkeys exhibit elevated circulating prolactin ( PRL ) which may be the result of enhanced pituitary responsiveness to vasoactive intestinal peptide ( P01282 ) . This hypothesis was tested by comparison of spontaneous and porcine P01282 -induced PRL secretion from anterior pituitary cells of hens in various reproductive conditions . The effect of P01282 and luteinizing hormone releasing hormone ( P01148 ) , alone and in combination , on luteinizing hormone ( LH ) secretion was also examined . Incubation with pVIP ( 10(-10) to 10(-6) M ) significantly stimulated PRL secretion at all incubation times tested ( 1-5 hr ) . This increase was greatest in cells from incubating hens , with those from laying , photorefractory , and quiescent ( nonphotostimulated ) hens secreting successively less PRL . These responses were obtained when spontaneous PRL secretions were compared . P01282 induced approximately a similar 1.5-fold increase in LH secretion , in all reproductive groups . Also , P01282 enhanced P01148 -induced LH secretion ( 1.2- to 1.6-fold ; P less than 0.0001 ) . It is concluded that PRL secretion in vitro by pituitary cells from turkey hens in various reproductive stages reflects the circulating levels of PRL at these stages . Circulating endothelial progenitor cells in acromegaly . BACKGROUND : Endothelial progenitor cells ( EPCs ) , involved in the repairing mechanisms of vascular damage , are positively correlated to insulin-like growth factor I ( P05019 ) concentrations in healthy adults . However , the levels of EPCs and their role in acromegalic patients have never been investigated . AIM : We conducted a cross-sectional study in order to assess the levels of the different phenotypes of circulating EPC in acromegalic patients . SUBJECTS AND METHODS : The study was performed at the Endocrinology Unit of Federico II University and at the Unit of Metabolic Diseases and Endocrinology of the Second University of Naples . Fifty-five acromegalic patients and 65 healthy controls were studied . EPCs were assessed by flow cytometry and P05019 by immunoradiometric assay . RESULTS : Compared with subjects of the control group , acromegalic patients showed significantly higher levels of EPCs phenotypes expressing P35968 antigen [ P35968 + , cells per 106 events , median and interquartile range , 44 ( 28-67 ) vs 23 ( 13-40 ) , p=0.006 ; P28906 + P35968 + 25 ( 18-38 ) vs 12 ( 8-17 ) , p < 0.001 ; CD133+ P35968 + 17 ( 13-30 ) vs 8 ( 6-12 ) , p < 0.001 ; P28906 + P35968 +CD133+ 16 ( 12-25 ) vs 8 ( 6-10 ) , p < 0.001 ] . There was a positive correlations between P28906 + P35968 +CD133+ cells count and P05019 in acromegaly group ( r=0.79 , p < 0.001 ) . CONCLUSIONS : Acromegalic patients show higher circulating EPCs levels expressing P35968 , positively correlated with P05019 , suggesting a role for P05019 in regulating the expression of this surface marker in the early phase of EPCs differentiation . Exogenous gonadotrophin-releasing hormone ( DB00644 ) stimulates LH secretion in male monkeys ( Macaca fascicularis ) treated chronically with high doses of a DB00644 antagonist . We reported previously that after a single injection of a gonadotrophin-releasing hormone ( DB00644 ) antagonist to male monkeys , exogenous DB00644 stimulated LH secretion in a time- and dose-dependent manner , indicating that DB00644 antagonist-induced blockade of LH secretion resulted from pituitary P30968 occupancy . The present study was performed to investigate whether DB00644 can also restore a blockade of LH and testosterone secretion during chronic DB00644 antagonist administration . Four adult male cynomolgus monkeys ( Macaca fascicularis ) received daily s.c. injections of the DB00644 antagonist [ N-Ac-D-pCl-Phe1,2,D- Q13507 ,D-Arg6-D-Ala10 ] - DB00644 ( ORG 30276 ) at a dose of 1400-1600 micrograms/kg for 8 weeks . Before the DB00644 antagonist was given and during weeks 3 and 8 of treatment , pituitary stimulation tests were performed with 0.5 , 5 , 50 and 500 micrograms synthetic DB00644 , administered in increasing order at intervals of 24 h . At 8 weeks , a dose of 1000 micrograms DB00644 was also given . All doses of DB00644 significantly ( P less than 0.05 ) stimulated serum concentrations of bioactive LH ( 3- to 8-fold ) and testosterone ( 2.6- to 3.8-fold ) before the initiation of DB00644 antagonist treatment . After 3 weeks of DB00644 antagonist treatment , only 50 and 500 micrograms DB00644 doses were able to increase LH and testosterone secretion . Release of LH was significantly ( P less than 0.05 ) more elevated with 500 micrograms compared with 50 micrograms DB00644 . After 8 weeks , only the highest dose of 1000 micrograms elicited a significant ( P less than 0.05 ) rise in LH secretion . Basal hormone levels just before the bolus injection of DB00644 were similar ( P greater than 0.10-0.80 ) . ( ABSTRACT TRUNCATED AT 250 WORDS ) TGF-beta-treated microglia induce oligodendrocyte precursor cell chemotaxis through the P14210 -c- DB00134 pathway . In acute experimental autoimmune encephalomyelitis ( EAE ) , demyelination is induced by myelin-specific P01730 (+) T lymphocytes and myelin-specific antibodies . Recovery from the disease is initiated by cytokines which suppress T cell expansion and the production of myelin-toxic molecules by macrophages . Th2/3 cell-derived signals may also be involved in central nervous system ( CNS ) repair . Remyelination is thought to be initiated by the recruitment and differentiation of oligodendrocyte precursor cells ( OPC ) in demyelinated CNS lesions . Here , we report that unlike Th1 cytokines ( P01375 , P01579 ) , the Th2/3 cytokine TGF-beta induces primary microglia from C57BL/6 mice to secrete a chemotactic factor for primary OPC . We identified this factor to be the hepatocyte growth factor ( P14210 ) . Our studies show that P01137 -2-3 as well as IFN-beta induce P14210 secretion by microglia and that antibodies to the P08581 c- DB00134 abrogate OPC chemotaxis induced by TGF-beta2-treated microglia . In addition we show spinal cord lesions in EAE induced in SJL/J mice to contain both OPC and P14210 producing macrophages in the recovery phase , but not in the acute stage of disease . Taken these findings , TGF-beta may play a pivotal role in remyelination by inducing microglia to release P14210 which is both a chemotactic and differentiation factor for OPC . 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 .

MH_train_1588

MH_train_1588

MH_train_1588

interacts_with DB04844?

multiple_choice

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 . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . 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 . Mesenchymal stem cells control alloreactive CD8(+) P10747 (-) T cells . P10747 / P33681 co-stimulation blockade with belatacept prevents alloreactivity in kidney transplant patients . However , cells lacking P10747 are not susceptible to belatacept treatment . As CD8(+) P10747 (-) T-cells have cytotoxic and pathogenic properties , we investigated whether DB05914 ( O60682 ) are effective in controlling these cells . In mixed lymphocyte reactions ( P08235 ) , O60682 and belatacept inhibited peripheral blood mononuclear cell ( PBMC ) proliferation in a dose-dependent manner . O60682 at O60682 /effector cell ratios of 1:160 and 1:2·5 reduced proliferation by 38·8 and 92·2 % , respectively . DB06681 concentrations of 0·1 μg/ml and 10 μg/ml suppressed proliferation by 20·7 and 80·6 % , respectively . Both treatments in combination did not inhibit each other 's function . Allostimulated CD8(+) P10747 (-) T cells were able to proliferate and expressed the cytolytic and cytotoxic effector molecules granzyme B , interferon ( IFN ) -γ and tumour necrosis factor ( P01375 ) -α . While belatacept did not affect the proliferation of CD8(+) P10747 (-) T cells , O60682 reduced the percentage of P10747 (-) T cells in the proliferating CD8(+) T cell fraction by 45·9 % ( P = 0·009 ) . CD8(+) P10747 (-) T cells as effector cells in P08235 in the presence of P01730 (+) T cell help gained P10747 expression , an effect independent of O60682 . In contrast , allostimulated P10747 (+) T cells did not lose P10747 expression in P08235 - O60682 co-culture , suggesting that O60682 control pre-existing P10747 (-) T cells and not newly induced P10747 (-) T cells . In conclusion , alloreactive CD8(+) P10747 (-) T cells that remain unaffected by belatacept treatment are inhibited by O60682 . This study indicates the potential of an O60682 -belatacept combination therapy to control alloreactivity . 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 . 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 . Distinct binding mode of multikinase inhibitor lenvatinib revealed by biochemical characterization . DB09078 is an oral multikinase inhibitor that selectively inhibits vascular endothelial growth factor ( P15692 ) receptors 1 to 3 and other proangiogenic and oncogenic pathway-related receptor tyrosine kinases . To elucidate the origin of the potency of lenvatinib in P15692 receptor 2 ( P35968 ) inhibition , we conducted a kinetic interaction analysis of lenvatinib with P35968 and X-ray analysis of the crystal structure of P35968 -lenvatinib complexes . Kinetic analysis revealed that lenvatinib had a rapid association rate constant and a relatively slow dissociation rate constant in complex with P35968 . Co-crystal structure analysis demonstrated that lenvatinib binds at its DB00171 mimetic quinoline moiety to the DB00171 binding site and to the neighboring region via a cyclopropane ring , adopting an DB00128 - DB00120 - DB00145 ( DFG ) - " in " conformation . These results suggest that lenvatinib is very distinct in its binding mode of interaction compared to the several approved P35968 kinase inhibitors . 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 . 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 . 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 . Native low-density lipoprotein increases endothelial cell nitric oxide synthase generation of superoxide anion . To examine mechanisms by which native low-density lipoprotein ( n-LDL ) perturbs endothelial cell ( EC ) release of superoxide anion ( O2- ) and nitric oxide ( NO ) , ECs were incubated with n-LDL at 240 mg cholesterol per deciliter for 4 days with media changes every 24 hours . n-LDL increases EC release of O2- by more than fourfold and increases nitrite production by 57 % . In the conditioned media from day-4 incubations , n-LDL increases total nitrogen oxides 20 times control EC ( C-EC ) levels . However , n-LDL did not alter EC NO synthase ( P29474 ) enzyme activity as measured by the [3H]citrulline assay . N omega-Nitro-L-arginine methyl ester , a specific inhibitor of P29474 activity , increases C-EC release of O2- by > 300 % but decreases LDL-treated EC ( LDL-EC ) release by > 95 % . L- DB00125 inhibits the release of O2- from LDL-ECs by > 95 % but did not effect C-EC release of O2- . Indomethacin and SKF 525A partially attenuate LDL-induced increases in O2- production by approximately 50 % and 30 % , respectively . Thus , n-LDL increases O2- and NO production , which increases the likelihood of the formation of peroxynitrite ( ONOO- ) , a potent oxidant . n-LDL increases the levels of nitrotyrosine , a stable oxidation product of ONOO- , and tyrosine by approximately 50 % . In spite of this increase in oxidative metabolism , analysis of thiobarbituric acid substances reveals that no significant changes in the oxidation of n-LDL occur during the 24-hour incubations with ECs. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . DB00998 : a selective type 1B/1D serotonin receptor agonist for the treatment of migraine headache . DB00998 belongs to an innovative family of compounds aimed at breaking through the long-standing barrier of migraine headache understanding and treatment . While a typology of headaches has been recognized for some time , and a number of therapies have been introduced for reduction of headache pain and duration , the causes of migraine remain a subject of debate . Those prone to attacks continue to endure them and suffer the related symptoms such as nausea and disorientation . DB00998 , like all the triptans , acts by inducing vasoconstriction of the meningeal arteries . It has been shown in pharmacological tests to act selectively as a potent agonist of serotonin P28222 /1D receptors . DB00998 has been well tolerated in humans and efficacious in reducing headache pain and duration in clinical trials , which have also indicated that dose adjustments for age or gender are not necessary for the drug . Patients have found the use of frovatriptan acceptable over the long-term , and overall a low-incidence of adverse effects has been reported . Though not a prophylactic , frovatriptan has demonstrated the potential to significantly improve the therapeutic approaches to the treatment of migraine . Joint effects of P29474 gene T-786C and P00325 Arg47His polymorphisms on the risk of premature coronary artery disease . INTRODUCTION : Both T-786C mutation in endothelial nitric oxide synthase ( P29474 ) gene and alcohol dehydrogenase ( DB00067 ) gene polymorphism such as P00326 gamma1/gamma2 have been reportedly associated with coronary artery disease ( CAD ) . Since P00325 Arg47His polymorphism is common in Asian population , the aim of this present study was to assess the interaction between P29474 gene T-786C and P00325 Arg47His polymorphisms on premature CAD risk . MATERIALS AND METHODS : Hospital-based case-control study was conducted with 167 premature CAD and 235 late-onset CAD patients . Polymerase chain reaction restriction fragment length polymorphism was used to detect the polymorphisms . Multivariate logistic regression model was performed to adjust the potential confounders and estimate odds ratios ( ORs ) with 95 % confidence intervals ( CIs ) . Synergy index ( S ) was the measure to assess the interaction as departure from additivity . RESULTS : After the adjustment for the potential confounders , and compared with the carriers of TT and DB00125 / DB00125 as the reference , the ORs with 95 % CIs in parentheses of premature CAD were that 1.13 ( 0.19-6.59 ) for CT or CC and DB00125 / DB00125 carriers ; 2.24 ( 0.77-6.49 ) for TT and DB00125 / DB00117 or DB00117 / DB00117 carriers ; 4.18 ( 1.32-13.22 ) for CT or CC and DB00125 / DB00117 or DB00117 / DB00117 carriers , respectively . Based on those ORs , S was 2.32 ( 95 % CI : 0.37-14.72 ) . CONCLUSIONS : The mutant genotypes of P29474 gene T-786C mutation and the fast form of P00325 Arg47His polymorphism had an additive interaction on the risk of premature CAD in Chinese population . Further investigations with big sample size are necessary for confirming this additive interaction . 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 . Profile of cabozantinib and its potential in the treatment of advanced medullary thyroid cancer . Medullary thyroid cancer is an uncommon malignancy for which until recently little effective treatment existed . It is often characterized by mutation and overexpression of the receptor tyrosine kinases P07949 ( rearranged during transfection ) , P35968 ( vascular endothelial growth factor receptor 2 ) and MET ( mesenchymal-epithelial transition factor ) , which make attractive targets for drug development . DB08875 is an orally bioavailable tyrosine kinase inhibitor which blocks MET , VEGRF2 and P07949 , and has shown considerable activity in medullary thyroid cancer in a Phase III trial , including in heavily pretreated patients . Its novel combination of vascular endothelial growth factor and MET inhibition is believed to address the MET escape pathway , which is thought to be the cause of nonsustained tumor responses resulting from inhibition of vascular endothelial growth factor alone . P00747 activator inhibitor type 1 ( P05121 ) is a valuable biomarker for predicting the metabolic syndrome ( MS ) in institutionalized elderly residents in Taiwan . Circulating levels of inflammatory and prothrombotic factors are elevated in the metabolic syndrome ( MS ) and linked with the occurrence of cardiovascular events . The aim of our study was to investigate the relationship between inflammatory and prothrombotic markers and the MS in elderly institutionalized residents . A total of 326 non-diabetic residents of Chuang-Hua Veterans Care Home ( age : 79.9+/-4.1 years ; 100 % males ) were enrolled . MS was diagnosed according to the DB00551 /NHLBI Scientific Statement criteria . Body fat percentage was measured by bioelectrical impedance analysis . P01308 resistance was calculated by homeostasis model assessment for insulin resistance ( HOMA-IR ) . Inflammatory markers , including tumor necrosis factor-a ( P01375 ) , high sensitivity P02741 ( hsCRP ) , and plasminogen activator inhibitor-1 ( P05121 ) , were determined using ELISA . Elderly residents with the MS had higher systolic and diastolic blood pressures ( both p < 0.001 ) and higher HOMA-IR ( p < 0.001 ) , hsCRP ( p = 0.008 ) , and P05121 levels ( p < 0.001 ) than those without the MS . On multivariate logistic regression analysis , P05121 was an independent risk factor for the MS . Of the MS components , elderly residents with higher waist circumferences and higher levels of plasma fasting glucose , and triglyceride ( TG ) , and lower levels of high density lipoprotein ( HDL ) had higher P05121 levels than those without the above components . Cl- DB05511 enhances P01375 -α release in peritoneal macrophages stimulated with LPS . P0DMS8 ( A3R ) belongs to the Gi/Gq-coupled receptor family , that leads to the intracellular DB02527 reduction and intracellular calcium increase , respectively . A3R is widely expressed and it can play a crucial role in many patho-physiological conditions , including inflammation . Here we investigate the effect of Cl- DB05511 , A3R agonist , on the production of P01375 -α . We found that Cl- DB05511 enhances LPS-induced P01375 -α release in peritoneal macrophages . This effect is reduced by MRS1191 , A3R antagonist and by forskolin , activator of adenylyl cyclase . pIκBα increased in LPS+Cl- DB05511 -treated macrophages , while total IκB kinase-β ( IKKβ ) reduced . Indeed , p65NF-κB nuclear translocation increased in cells treated with LPS+Cl- DB05511 . Moreover , IMD 0354 , IKKβ inhibitor , significantly abrogated the effect of Cl- DB05511 on P01375 -α release . Inhibition of protein kinase C ( PKC ) significantly reduced Cl- DB05511 -induced P01375 -α release in LPS-stimulated macrophages . Furthermore , LY-294002 , PI3K inhibitor , reduced the P01375 -α production enhanced by Cl- DB05511 , although the phosphorylation status of Akt did not change in cells treated with LPS+Cl- DB05511 than LPS alone . In summary , these data show that Cl- DB05511 is able to enhance P01375 -α production in LPS-treated macrophages in an NF-κB- dependent manner . DB11320 contributes to tissue remodeling via periostin expression . DB11320 is thought to have a critical role in the synthesis of extracellular matrix in skin and may be involved in tissue remodeling of allergic diseases . Recent studies revealed that periostin , a matricelluar protein , contributed to tissue remodeling ; however , a link between periostin and histamine remains unproven . We investigated whether periostin was involved in histamine-induced collagen production . Cultured dermal fibroblasts derived from wild-type ( WT ) or periostin knockout ( PN(-/-) ) mice were stimulated with histamine , and then collagen and periostin production was evaluated . DB11320 induced collagen gene expression in WT fibroblasts in the late phase but not in the early phase , whereas no effect on collagen expression was observed in histamine-stimulated PN(-/-) fibroblasts . In WT fibroblasts , histamine directly induced periostin expression in a dose-dependent manner , and an H1 receptor antagonist blocked both periostin and collagen expression . DB11320 activated extracellular signal-regulated kinase 1/2 ( P27361 /2 ) through the H1 receptor . Q15063 induction was inhibited by either H1 antagonist or P27361 /2 inhibitor treatment in vitro and was attenuated in P35367 (-/-) mice . Elevated expression of periostin was found in lesional skin from atopic dermatitis patients . These results suggest that histamine mediates periostin induction and collagen production through activation of the H1 receptor-mediated P27361 /2 pathway ; furthermore , histamine may accelerate the chronicity of atopic dermatitis . Serotonergic regulation of somatosensory cortical development : lessons from genetic mouse models . Monoaminergic neurotransmitter systems appear early during embryogenesis , suggesting that they could play important roles in brain development . Accumulated evidence indicates that serotonin ( 5-hydroxytryptamine , 5-HT ) regulates neural as well as nonneural development , including early aspects of embryonic development , differentiation of neuronal progenitors , and morphogenesis of the craniofacial region , heart and limb . Recent studies using monoamine oxidase-A ( P21397 ) , 5-HT transporter , vesicular monoamine transporter-2 ( Q05940 ) and P28222 receptor single , double and triple knockout mice have provided evidence that the serotonergic system plays important roles in barrel field formation in the developing somatosensory cortex . Here we review evidence from these genetic mouse models and , based on the accumulated evidence , propose a testable model for future studies of mechanisms underlying serotonergic regulation of cortical development . Q5TCZ1 -mapping comparison between river buffalo chromosome 7 and sheep chromosome 6 : assignment of new loci and comparison with HSA4 . Synchronized peripheral blood lymphocytes from both river buffalo ( BBU ) and sheep ( OAR ) were treated for late incorporation of both BrdU and H-33258 to obtain R-banded preparations to be used for Q5TCZ1 -mapping . Ovine BAC-clones were hybridized for three days on slides pre-exposed to UV light after H-33258 staining . The following loci were mapped : Q96P65 ( BBU7q13 , OAR6q13 ) , Q8N609 (OAR6q13dist) , Q08209 ( BBU7q21 , OAR6q15 ) , P37840 ( OAR6q17 ) , Q9UBK2 ( BBU7q23 , OAR6q17 ) , O60701 ( BBU7q25prox , OAR6q22prox ) , P35968 ( BBU7q27 , OAR6q22 ) , Q96LI5 ( BBU7q32prox , OAR6q32prox ) , Q7Z3B4 ( BBU7q32 , BBU6q32 ) , DMP1 ( BBU7q34dist-q36prox , OAR6q34dist-q36prox ) , P09417 ( BBU7q36 , OAR6q36 ) . All loci mapped in homoeologous chromosomes and chromosome bands of the two species and their locations are in agreement with the previous RH-mapping performed on BBU7 with some difference in the distal region of BBU7 . However , the present cytogenetic map better anchors the RH-map on specific river buffalo chromosome bands . In addition , eleven loci were assigned for the first time in sheep to OAR6 , noticeably extending the cytogenetic map on this important chromosome which encodes caseins . Two loci ( Q8N609 and P37840 ) mapped in sheep were unmapped in river buffalo in three different Q5TCZ1 experiments . Comparisons between integrated cytogenetic maps of BBU7/OAR6 ( and BTA6 ) with human chromosome 4 ( HSA4 ) revealed complex chromosome rearrangements differentiating these chromosomes . P00747 activator is involved in the hCG-induced neutrophil extravasation and vasopermeability increase in the rat testis . The role of proteolytic enzymes in the hCG-induced increase in testicular vasopermeability and neutrophil extravasation was studied using protease inhibitors . An intra-testicular injection of hCG together with incubation medium conditioned by polymorphonuclear leucocytes ( PMNs ) caused a significant increase in vasopermeability and a coincident extravasation of PMN 's from the postcapillary venules in the rat testis . When p-aminobenzamidine , a serine protease inhibitor which inhibits urokinase-type plasminogen activator , was administered together with hCG in the incubation medium , both the permeability increase and PMN extravasation were prevented . DB06692 , another serine protease inhibitor , and Eglin C , a specific neutrophil elastase and cathepsin G inhibitor were , however , without effect . None of these inhibitors caused any non-specific vascular effects in the testis at the concentrations used . These results support the concept that the hCG-induced increase in vasopermeability in the rat testis is related to extravasation of PMNs and suggest that urokinase-type plasminogen activator is involved in migration of these cells through the postcapillary venular walls .

MH_train_1589

MH_train_1589

MH_train_1589

interacts_with DB01418?

multiple_choice

Role of endothelial cells in antihyperalgesia induced by a DB00669 and β-blocker . While blood vessels have long been implicated in diverse pain syndromes ( e.g. , migraine headache , angina pectoris , vasculitis , and Raynaud 's syndrome ) , underlying mechanisms remain to be elucidated . Recent evidence supports a contribution of the vascular endothelium in endothelin-1-induced hyperalgesia , and its enhancement by repeated mechanical stimulation ; a phenomenon referred to as stimulus-induced enhancement of ( endothelin ) hyperalgesia ( SIEH ) . SIEH is thought to be mediated by release of DB00171 from endothelial cells , to act on P56373 receptors on nociceptors . In the present study we evaluated the ability of another vasoactive hyperalgesic agent , epinephrine , to induce endothelial cell-dependent hyperalgesia and SIEH . We found that epinephrine also produces hyperalgesia and SIEH . Both P56373 receptor antagonists , A317491 and octoxynol-9 , which attenuate endothelial cell function , eliminated SIEH without affecting epinephrine hyperalgesia . We further evaluated the hypothesis that members of two important classes of drugs used to treat migraine headache , whose receptors are present in endothelial cells - the triptans and β blockers - have a vascular component to their anti-hyperalgesic action . For this , we tested the effect of ICI-118,551 , a β₂-adrenergic receptor antagonist and sumatriptan , an agonist at P28222 and 5-HT₁D receptors , on nociceptive effects of endothelin and epinephrine . ICI-118,551 inhibited endothelin SIEH , and attenuated epinephrine hyperalgesia and SIEH . Sumatriptan inhibited epinephrine SIEH and inhibited endothelin hyperalgesia and SIEH , while having no effect on epinephrine hyperalgesia or the hyperalgesia induced by a prototypical direct-acting inflammatory mediator , prostaglandin E₂ . These results support the suggestion that triptans and β-blockers interact with the endothelial cell component of the blood vessel to produce anti-hyperalgesia . Advances in Q9BWK5 assessment of gliomas and response to anti- P15692 therapy . DB00112 is thought to normalize tumor vasculature and restore the blood-brain barrier , decreasing enhancement and peritumoral edema . Conventional measurements of tumor response rely upon dimensions of enhancing tumor . After bevacizumab treatment , glioblastomas are more prone to progress as nonenhancing tumor . The RANO ( Response Assessment in Neuro-Oncology ) criteria for glioma response use fluid-attenuated inversion recovery ( FLAIR ) / P24752 hyperintensity as a surrogate for nonenhancing tumor ; however , nonenhancing tumor can be difficult to differentiate from other causes of FLAIR/ P24752 hyperintensity ( e.g. , radiation-induced gliosis ) . Due to these difficulties , recent efforts have been directed toward identifying new biomarkers that either predict treatment response or accurately measure response of both enhancing and nonenhancing tumor shortly after treatment initiation . This will allow for earlier treatment decisions , saving patients from the adverse effects of ineffective therapies while allowing them to try alternative therapies sooner . An active area of research is the use of physiologic imaging , which can potentially detect treatment effects before changes in tumor size are evident . DB00315 , a new central and peripherally acting P28221 receptor agonist in the acute oral treatment of migraine : a double-blind , placebo-controlled , dose-range finding study . DB00315 is a novel , centrally and peripherally , acting 5-hydroxytryptamine1D receptor agonist . We investigated the efficacy and safety of 1 , 5 , and 25 mg of oral DB00315 in the acute treatment of migraine in a randomized , double-blind , placebo-controlled , parallel-group clinical trial involving 84 patients . The proportion of patients in whom the headache improved within 2 hours from moderate or severe to mild or no pain ( primary efficacy measure ) was 15 % for placebo-treated patients and 27 % ( 1 mg ) , 62 % ( 5 mg ) , and 81 % ( 25 mg ) for patients treated with DB00315 . Treatment differences compared with placebo were 12 % ( 95 % CI - 12 , 37 ; p = 0.460 ) for 1 mg DB00315 , 47 % ( CI 21 , 73 ; p < 0.005 ) for 5 mg DB00315 , and 66 % ( CI 43,89 ; p < 0.001 ) for 25 mg DB00315 . Photophobia and nausea also showed improvement after DB00315 . Adverse events were generally mild and transient in all treatment groups . There were no clinically significant changes in ECG recordings , blood pressure , or laboratory tests . Oral DB00315 ( 5 and 25 mg ) is highly effective and well tolerated in the acute treatment of migraine . The response rates and treatment differences compared with placebo in this study suggest possible superiority over existing antimigraine therapies . This needs to be confirmed in formal comparative trials . 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 . 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 . Regulatory roles of sex hormones in cutaneous biology and immunology . Recent studies have revealed that sex hormones manifest a variety of biological and immunological effects in the skin . Pregnancy , menstruation and the menopause modulate the natural course of psoriasis , indicating a female hormone-induced regulation of skin inflammation . Estrogen in vitro down-regulates the production of the neutrophil , type 1 T cell and macrophage-attracting chemokines , P10145 , P02778 , P13501 , by keratinocytes , and suppresses IL-12 production and antigen-presenting capacity while enhancing anti-inflammatory P22301 production by dendritic cells . These data indicate that estrogen may attenuate inflammation in psoriatic lesions . Estrogen , alone or together with progesterone , prevents or reverses skin atrophy , dryness and wrinkles associated with chronological or photo-aging . Estrogen and progesterone stimulate proliferation of keratinocytes while estrogen suppresses apoptosis and thus prevents epidermal atrophy . Estrogen also enhances collagen synthesis , and estrogen and progesterone suppress collagenolysis by reducing matrix metalloproteinase activity in fibroblasts , thereby maintaining skin thickness . Estrogen maintains skin moisture by increasing acid mucopolysaccharide or hyaluronic acid levels in the dermis . Progesterone increases sebum secretion . Estrogen accelerates cutaneous wound healing stimulating P01138 production in macrophages , GM- P04141 production in keratinocytes and P09038 and TGF-beta1 production in fibroblasts , leading to the enhancement of wound re-innervation , re-epithelialization and granulation tissue formation . In contrast , androgens prolong inflammation , reduce deposition of extracellular matrix in wounds , and reduce the rate of wound healing . Estrogen enhances P15692 production in macrophages , an effect that is antagonized by androgens and which may be related to the development of granuloma pyogenicum during pregnancy . These regulatory effects of sex steroids may be manipulated as therapeutic or prophylactic measures in psoriasis , aging , chronic wounds or granuloma pyogenicum . P30556 antagonist attenuates LPS-induced acute lung injury . Angiotensin II is able to trigger inflammatory responses through an angiotensin II type 1 ( AT1 ) receptor . The role of AT1 receptor in acute lung injury ( ALI ) is poorly understood . Mice were randomly divided into three groups ( n=40 each groups ) : NS group ; LPS group ( 2mg/kg LPS intratracheally ) ; and LPS+ZD 7155 group , 10mg/kg ZD 7155 ( an AT1 receptor antagonist ) intraperitoneally 30 min prior to LPS exposure . Samples from the lung were isolated and assayed for histopathology analyses or proinflammatory gene expressions , angiotensin II receptors expressions and nuclear factors activities . LPS exposure resulted in severe ALI , elevated levels of P01375 and P01584 mRNA expressions , and increased activities of NF-kappaB and activated protein ( AP ) -1 . Upregulation of AT1 receptor and down-regulation of P50052 receptor were also observed after LPS challenge . Pretreatment with ZD 7155 significantly inhibited the increase of AT1 receptor expression and upregulated P50052 receptor expression . ZD 7155 also reduced the mRNA expression of P01375 and P01584 , inhibited the activation of NF-kappaB and AP-1 , and improved lung histopathology . These findings suggest that antagonism of AT1 receptor inhibits the activation of NF-kappaB and AP-1 in the lung , which may mediate the release of P01375 and P01584 and contribute to LPS-induced ALI . 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 . Granulocyte-colony stimulating factor ( DB00099 ) induces mechanical hyperalgesia via spinal activation of Q96HU1 kinases and PI3K in mice . Granulocyte-colony stimulating factor ( DB00099 ) is a current pharmacological approach to increase peripheral neutrophil counts after anti-tumor therapies . Pain is most relevant side effect of G- P04141 in healthy volunteers and cancer patients . Therefore , the mechanisms of G- P04141 -induced hyperalgesia were investigated focusing on the role of spinal mitogen-activated protein ( Q96HU1 ) kinases P29323 ( extracellular signal-regulated kinase ) , JNK ( Jun N-terminal Kinase ) and p38 , and PI(3)K ( phosphatidylinositol 3-kinase ) . G- P04141 induced dose ( 30-300 ng/paw ) -dependent mechanical hyperalgesia , which was inhibited by local post-treatment with morphine . This effect of morphine was reversed by naloxone ( opioid receptor antagonist ) . Furthermore , G- P04141 -induced hyperalgesia was inhibited in a dose-dependent manner by intrathecal pre-treatment with P29323 ( PD98059 ) , JNK ( SB600125 ) , p38 ( SB202190 ) or PI(3)K ( wortmanin ) inhibitors . The co-treatment with Q96HU1 kinase and PI(3)K inhibitors , at doses that were ineffective as single treatment , significantly inhibited G- P04141 -induced hyperalgesia . Concluding , in addition to systemic opioids , peripheral opioids as well as spinal treatment with Q96HU1 kinases and PI(3)K inhibitors also reduce G- P04141 -induced pain . 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 . Inhibition of the striatum-enriched phosphodiesterase Q9Y233 : a novel approach to the treatment of psychosis . Phosphodiesterase 10A ( Q9Y233 ) is a recently identified cyclic nucleotide phosphodiesterase expressed primarily in dopaminoreceptive medium spiny neurons of the striatum . We report that papaverine is a potent , specific inhibitor of Q9Y233 and use this compound to explore the role of Q9Y233 in regulating striatal function . DB01113 administration produces an increase in striatal tissue levels of cGMP and an increase in extracellular DB02527 measured by microdialysis . These cyclic nucleotide changes are accompanied by increases in the phosphorylation of CREB and P29323 , downstream markers of neuronal activation . In rats , papaverine potentiates haloperidol-induced catalepsy , consistent with the hypothesis that inhibition of Q9Y233 can increase striatal output and prompting a further evaluation of papaverine in models predictive of antipsychotic activity . DB01113 is found to inhibit conditioned avoidance responding in rats and mice and to inhibit PCP- and amphetamine-stimulated locomotor activity in rats . The effects of papaverine on striatal cGMP and CREB and P29323 phosphorylation , as well as on conditioned avoidance responding , were absent in Q9Y233 knockout mice , indicating that the effects of the compound are the result of Q9Y233 inhibition . These results indicate that Q9Y233 regulates the activation of striatal medium spiny neurons through effects on DB02527 - and cGMP-dependent signaling cascades . Furthermore , the present results demonstrate that papaverine has efficacy in behavioral models predictive of antipsychotic activity . Thus , inhibition of Q9Y233 may represent a novel approach to the treatment of psychosis . 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 . Q14703 promotes murine progenitor cell egress and mobilization via P21453 -mediated ROS signaling and P48061 release . The mechanisms of hematopoietic progenitor cell egress and clinical mobilization are not fully understood . Herein , we report that in vivo desensitization of DB03203 -1-phosphate ( Q14703 ) receptors by FTY720 as well as disruption of Q14703 gradient toward the blood , reduced steady state egress of immature progenitors and primitive Sca-1(+)/c-Kit(+)/Lin(-) ( SKL ) cells via inhibition of P48061 release . Administration of DB06809 or G- P04141 to mice with deficiencies in either Q14703 production or its receptor Q14703 (1) , or pretreated with FTY720 , also resulted in reduced stem and progenitor cell mobilization . Mice injected with DB06809 or G- P04141 demonstrated transient increased Q14703 levels in the blood mediated via P42345 signaling , as well as an elevated rate of immature c-Kit(+)/Lin(-) cells expressing surface Q14703 (1) in the bone marrow ( BM ) . Importantly , we found that Q14703 induced P48061 secretion from BM stromal cells including P48681 (+) DB05914 via reactive oxygen species ( ROS ) signaling . Moreover , elevated ROS production by hematopoietic progenitor cells is also regulated by Q14703 . Our findings reveal that the Q14703 / Q14703 (1) axis regulates progenitor cell egress and mobilization via activation of ROS signaling on both hematopoietic progenitors and BM stromal cells , and P48061 release . The dynamic cross-talk between Q14703 and P48061 integrates BM stromal cells and hematopoeitic progenitor cell motility . Purification and characterization of mouse O15528 overproduced by an Escherichia coli system coexpressing molecular chaperonins GroEL/ES . The expression of mouse O15528 in Escherichia coli has been dramatically enhanced by coexpression of GroEL/ES . To reveal the enzymatic properties of O15528 , we measured its hydroxylation activity toward vitamin D3 and DB01436 ( 1alpha(OH)D3 ) in addition to the physiological substrate DB00146 . Surprisingly , O15528 converted vitamin D3 to 1alpha, DB00146 . Both 1alpha-hydroxylation activity toward vitamin D3 , and 25-hydroxylation activity toward 1alpha(OH)D3 were observed . The Km and Vmax values for 25-hydroxylation activity toward 1alpha(OH)D3 were estimated to be 1.7 microM and 0.51 mol/min/mol P450 , respectively , while those for 1alpha-hydroxylation activity toward DB00146 were 0.050 microM and 2.73 mol/min/mol P450 , respectively . Note that the substrate must be fixed in the opposite direction in the substrate-binding pocket of O15528 between 1alpha-hydroxylation and 25-hydroxylation . Based on these results and the fact that human Q02318 and Streptomyces CYP105A1 also convert vitamin D3 to 1alpha, DB00146 , 1alpha-hydroxylation , and 25-hydroxylation of vitamin D3 appear to be closely linked together . [ Additive antiproteinuric effect of angiotensin II receptor antagonist and angiotensin-converting enzyme inhibitor in patients with chronic glomerulonephritis ] . P30556 blocker ( ARB ) and angiotensin-converting enzyme inhibitor ( ACEI ) have been thought to be effective for reducing proteinuria in patients with chronic glomerulonephritis . Recently , an additive effect of these two types of angiotensin blockers has been reported in patients with IgA nephropathy , but the mechanism responsible for the effect has not yet been determined . In this study , we examined additive effect of these two drugs in chronic glomerulonephritis patients . Ten patients with biopsy-proven primary glomerulonephritis ( eight IgA nephropathy patients , two membranous nephropathy patients ) , non-nephrotic proteinuria ( protein , 0.5 to 3.5 g/day ) received candesartan cilexetil ( 2 or 4 mg ) for 8 weeks . After the 8 weeks , a combination of perindopril erbumine ( 1 or 2 mg ) and candesartan cilexetil was administered to the patients . DB00790 was stopped after the 8-week administration of the two drugs . DB00796 alone reduced proteinuria by 13 % . Combination of these two drugs induced a more remarkable reduction of proteinuria ( 48 % ; p < 0.05 vs other periods ) . The decrease in mean blood pressure by the combination therapy was significantly correlated with the decrease in proteinuria . The combination of drugs also reduced the amount of urinary type-IV collagen excretion . An additive effect of ACEI and ARB on proteinuria and urinary type-IV collagen excretion was recognized in patients with chronic glomerulonephritis . 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 . Synthesis and evaluation of fluorinated fingolimod ( FTY720 ) analogues for sphingosine-1-phosphate receptor molecular imaging by positron emission tomography . DB03203 -1-phosphate ( Q14703 ) is a lysophospholipid that evokes a variety of biological responses via stimulation of a set of cognate G-protein coupled receptors ( GPCRs ) : P21453 - Q9H228 . Q14703 and its receptors ( S1PRs ) play important roles in the immune , cardiovascular , and central nervous systems and have also been implicated in carcinogenesis . Recently , the Q14703 analogue DB08868 ( FTY720 ) has been approved for the treatment of patients with relapsing multiple sclerosis . This work presents the synthesis of various fluorinated structural analogues of FTY720 , their in vitro and in vivo biological testing , and their development and application as [(18)F]radiotracers for the study of S1PR biodistribution and imaging in mice using small-animal positron emission tomography ( PET ) . DB01064 inhibits fibroblast growth factor-2-induced growth of renal epithelial cells . The signal transduction pathways modulating P09038 effects in renal tubular epithelial cells ( RTEc ) are not completely understood . Since the DB02527 and the mitogen-activated protein kinase ( MAPK ) pathways can modulate the growth of RTEc , we studied whether two DB02527 elevating agents , isoproterenol and 8-bromo- DB02527 , would modulate basic fibroblast growth factor ( P09038 ) induction of MAPK activity ( P28482 ) and cell proliferation in human renal proximal tubular epithelial cells ( RPTEc ) and Madin-Darby canine kidney cells ( MDCK clone EI1 ) . DB01064 , but not P09038 , stimulated DB02527 production in RPTEc and MDCKEI1 cells . P09038 , isoproterenol , and 8-bromo- DB02527 alone increased P28482 activity in both cell types . However , isoproterenol and 8-bromo- DB02527 partially inhibited the P09038 induction of P28482 activity , but only isoproterenol inhibited the proliferation of both cell types . PD098059 ( 25 microM ) , an inhibitor of MAPK kinase ( Q02750 /2 ) , blocked the P09038 mitogenic effects , but did not affect the 8-bromo- DB02527 -induced mitogenic effects in MDCKEI1 cells . These findings suggest that activation of P28482 is required but not sufficient for mitogenesis in RTEc . We conclude that isoproterenol inhibits the growth-promoting effects of P09038 in RTEc via MEK-dependent and -independent pathways . 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 . 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 . Mapping of the serotonin P28221 beta autoreceptor gene on chromosome 6 and direct analysis for sequence variants . Abnormal brain serotonin function may be characteristic of several neuropsychiatric disorders . Thus , it is important to identify polymorphic genes and screen for functional variants at loci coding for genes that control normal serotonin functions . P28221 beta is a terminal serotonin autoreceptor which may play a role in regulating serotonin synthesis and release . Using an SSCP technique we screened for P28221 beta coding sequence variants in psychiatrically interviewed populations , which included controls , alcoholics , and alcoholic arsonists and alcoholic violent offenders with low P04141 concentrations of the main serotonin metabolite 5-HIAA . A common polymorphism was identified in the P28221 beta gene with allele frequencies of 0.72 and 0.28 . The SSCP variant was caused by a silent G to C substitution at nucleotide 861 of the coding region . This polymorphism could also be detected as a HincII RFLP of amplified DNA . DNAs from informative CEPH families were typed for the HincII RFLP and analyzed with respect to 20 linked markers on chromosome 6 . Multipoint analysis placed the P28221 beta receptor gene between markers D6S286 and D6S275 . A maximum two-point lod score of 10.90 was obtained to D6S26 , which had been previously localized on 6q14-15 . Chromosomal aberrations involving this region have been previously shown to cause retinal anomalies , developmental delay , and abnormal brain development . This region also contains the gene for North Carolina-type macular dystrophy . The function of the selective inhibitors of cyclooxygenase 2 . Cyclooxygenase plays a pivotal role in the transformation of the arachidonic acid to prostaglandins ( PGs ) and thromboxane . It is composed of two kinds of enzymes , namely cyclooxygenase 1 ( P23219 ) and cyclooxygenase 2 ( P35354 ) . Cyclooxygenase 1 is the constructive enzyme whereas the cyclooxygenase 2 is the inducible enzyme . Inhibiting cyclooxygenase 1 is always associated with some undesirable side-effects , while inhibiting cyclooxygenase 2 can result in therapeutic effect . This has led the researchers to strive for searching the selective inhibitors inhibiting the P35354 instead of P23219 . It is very well known that pain and inflammation are alleviated through the inhibition of P35354 inhibitors such as DB00945 , which has resulted in the recent years , in the emergence of a range of P35354 inhibitors . Moreover , while evaluating the functions of the P35354 inhibitions , their significant role in treating glaucoma , preventing and suppressing cancer through their inhibitory activity was clearly revealed and many studies further demonstrated that P35354 is not only related to the inflammation of peripheral tissues but also to the inflammation manifested in the central nervous system . In addition , the nervous disorders also found an effective treatment with the administration of P35354 inhibitors . The above-mentioned findings delineate the role of the P35354 inhibitors as promising agents to be exploited in the treatment of many illnesses . This review will elucidate the functions of the P35354 inhibitors briefly and introduce some common selective inhibitors of P35354 .

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Activated lymphoid cells in human gliomas : morphofunctional and cytochemical evidence . To study defensive infiltrating cells , smear preparation from 20 fresh gliomas and autologous normal peritumoural brain tissue , used as control , were analysed . May-Grünwald Giemsa staining and cytochemical reaction markers of cellular functions [ acid phosphatase ( AP ) , dihydrofolate reductase ( P00374 ) , dipeptidilpeptidase ( DAP IV ) and serine esterase Q15722 -dependent ( SE ) ] , were employed to characterize the cells . The extent of the leukocytic infiltration and the percentage of activated lymphocytes ( " hand mirror " shape lymphocytes : DB00253 ; lymphocytes binding tumor : LBT ) were also studied . In addition serum levels of T cell growth factor interleukin-2 ( P60568 ) and of the soluble P60568 receptor ( sIL-2R ) were determined in the affected patients . In tumoural imprints , mostly in astrocytomas , we observed an increased percentage of P00374 and DAP IV positive lymphocytes , of DB00253 lymphocytes and of lymphocytes binding tumoral cells . Serum levels of P60568 were significantly increased in all patients whilst sIL-2R levels , were high only in glioblastoma . These findings indicate that in malignant gliomas there is stimulation of the immune system as witnessed by the presence of activated cells inside tumor tissue and soluble activating factors in serum . Pharmacological examination of the neurokinin-1 receptor mediating relaxation of human intralobar pulmonary artery . The effect of selective tachykinin receptor agonists and antagonists on human isolated intralobar pulmonary arterial rings was investigated . Neither Substance P ( SP ) nor neurokinin A ( P20366 ) contracted the arteries . Both of these agonists , however , were potent and efficacious at relaxing the arteries that were precontracted with phenylephrine . The negative log ( M ) EC(50) values for SP and P20366 were 9.0 and 8.3 , respectively . The neurokinin ( NK ) -3 selective agonist , senktide-analog , and the P21452 selective agonist , [beta-Ala(8)] P20366 (4-10) , caused neither contractions nor relaxations of the arteries , whereas the P25103 agonist Ac- [ Arg6 , Sar9 , DB00134 (O2)11 ] SP(6-11) ( P17405 -SP ) relaxed the tissue with a potency similar to SP . The relaxations to SP , P20366 , and P17405 -SP were competitively antagonized by the selective P25103 antagonist CP 99994 , with a pK(b) in the nanomolar range . Antagonism of the P17405 -SP-induced relaxations was also noted with FK 888 , RP 67580 , and L 732,138 , although these antagonists were much less potent than CP 99994 in this regard . Another P25103 selective antagonist , DB05790 , caused an insurmountable antagonism of the SP-induced relaxations . The P25103 -mediated relaxations could be blocked by removing the endothelium , or by a combination of N-nitro-L-arginine and indomethacin . Measurement of prostanoid generation revealed that in endothelial-intact but not endothelial-denuded tissue , P17405 -SP caused a selective increase in the production of 6-keto-PGF1alpha , the stable metabolite of prostacyclin . The results indicate that stimulation of NK-1 receptors leads to relaxation of human intralobar pulmonary arteries , which is mediated largely by nitric oxide and prostacyclin released from the endothelium of these vessels . Differential expression and function of phosphodiesterase 4 ( DB05876 ) subtypes in human primary P01730 + T cells : predominant role of Q08499 . Type 4 phosphodiesterases ( DB05876 ) are critical regulators in TCR signaling by attenuating the negative constraint of DB02527 . In this study , we show that anti-CD3/ P10747 stimulation of human primary P01730 (+) T cells increases the expression of the DB05876 subtypes P27815 , Q07343 , and Q08499 in a specific and time-dependent manner . P27815 and Q08499 mRNAs as well as enzyme activities were up-regulated within 5 days , Q07343 showed a transient up-regulation with highest levels after 24 h . The induction was shown to be independent of different stimulation conditions and was similar in naive and memory T cell subpopulations . To elucidate the functional impact of individual DB05876 subtypes on T cell function , we used DB05876 subtype-specific short-interfering RNAs ( siRNAs ) . Knockdown of either Q07343 or Q08499 inhibited P60568 release 24 h after stimulation ( time point of maximal P60568 concentrations ) to an extent similar to that observed with the panPDE4 inhibitor RP73401 ( piclamilast ) . Substantial amounts of P01579 or P05113 were measured only at later time points . siRNA targeting Q08499 showed a predominant inhibitory effect on these cytokines measured after 72 h . However , the inhibition of all cytokines was most effective when DB05876 siRNAs were applied in combination . Although the effect of DB05876 inhibition on T cell proliferation is small , the Q08499 -targeting siRNA alone was as effective as the panPDE4 inhibitor , whereas P27815 or Q07343 siRNAs had hardly an effect . In summary , individual DB05876 subtypes have overall nonredundant , but complementary , time-dependent roles in propagating various T cell functions and Q08499 is the form likely playing a predominant role . Exenatide does not evoke pancreatitis and attenuates chemically induced pancreatitis in normal and diabetic rodents . The risk of developing pancreatitis is elevated in type 2 diabetes and obesity . Cases of pancreatitis have been reported in type 2 diabetes patients treated with P0C6A0 ( P43220 ) receptor agonists . To examine whether the P43220 agonist exenatide potentially induces or modulates pancreatitis , the effect of exenatide was evaluated in normal or diabetic rodents . Normal and diabetic rats received a single exenatide dose ( 0.072 , 0.24 , and 0.72 nmol/kg ) or vehicle . Diabetic ob/ob or HF- Q11206 mice were infused with exenatide ( 1.2 and 7.2 nmol·kg(-1)·day(-1) ) or vehicle for 4 wk . Post-exenatide treatment , pancreatitis was induced with caerulein ( CRN ) or sodium taurocholate ( ST ) , and changes in plasma amylase and lipase were measured . In ob/ob mice , plasma cytokines ( IL-1β , P60568 , P05231 , P13500 , IFNγ , and TNFα ) and pancreatitis-associated genes were assessed . Pancreata were weighed and examined histologically . Exenatide treatment alone did not modify plasma amylase or lipase in any models tested . Exenatide attenuated CRN-induced release of amylase and lipase in normal rats and ob/ob mice but did not modify the response to ST infusion . Plasma cytokines and pancreatic weight were unaffected by exenatide . Exenatide upregulated Reg3b but not Il6 , Ccl2 , Nfkb1 , or Vamp8 expression . Histological analysis revealed that the highest doses of exenatide decreased CRN- or ST-induced acute inflammation , vacuolation , and acinar single cell necrosis in mice and rats , respectively . Ductal cell proliferation rates were low and similar across all groups of ob/ob mice . In conclusion , exenatide did not modify plasma amylase and lipase concentrations in rodents without pancreatitis and improved chemically induced pancreatitis in normal and diabetic rodents . Discovery of selective Q07343 inhibitors . In this study the first Q07343 selective inhibitor is described . Optimization of lead 2-arylpyrimidine derivatives afforded a series of potent Q07343 inhibitors with > 100-fold selectivity over the Q08499 isozyme . With a good pharmacokinetic profile , a selected compound exhibited potent anti-inflammatory effects in vivo and showed less emesis compared with DB03849 . DB01373 -sensing receptor activation in chronic kidney disease : effects beyond parathyroid hormone control . Secondary hyperparathyroidism ( SHPT ) is an important complication of advanced chronic kidney disease ( CKD ) . DB01012 , an allosteric modulator of the calcium-sensing receptor ( P41180 ) expressed in parathyroid glands , is the only calcimimetic approved to treat SHPT in patients on dialysis . By enhancing P41180 sensitivity for plasma extracellular calcium ( Ca(2+)0 ) , cinacalcet reduces serum parathyroid hormone , Ca(2+)0 , and serum inorganic phosphorous concentrations , allowing better control of SHPT and CKD-mineral and bone disorders . Of interest , the P41180 also is expressed in a variety of tissues where its activation regulates diverse cellular processes , including secretion , apoptosis , and proliferation . Thus , the existence of potential off-target effects of cinacalcet can not be neglected . This review summarizes our current knowledge concerning the potential role(s) of the P41180 expressed in various tissues in CKD-related disorders , independently of parathyroid hormone control . The future use of liraglutide : implications of the LEAD-2 study for treatment guidelines in type 2 diabetes . The effective identification and management of type 2 diabetes ( T2D ) in primary care is a healthcare priority . New antidiabetic agents , including glucagon-like peptide ( GLP ) -1 receptor agonists , may help overcome drawbacks with current treatments . These new agents have been reviewed in the updated National Institute for Health and Clinical Excellence ( NICE ) guidelines for the treatment of T2D . DB06655 , a P43220 agonist , was licensed for use in patients with T2D after the development of the NICE guidelines . Data from Phase III trials evaluating liraglutide are presented here in the context of the role of P43220 agonists in NICE guidelines . 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 . Chronic methamphetamine exposure alters immune function in normal and retrovirus-infected mice . Methamphetamine ( MA ) abuse represents a growing problem in the USA with an increase of sudden death . To evaluate the immune function alterations due to chronic methamphetamine use , we examined C57BL/C mice with LP-BM5 retrovirus infection plus methamphetamine exposure . Mice were randomly assigned to the following groups : placebo , placebo retrovirus-infected , uninfected MA treated and retrovirus-infected MA treated . Placebo , MA-treated groups were intraperitoneally injected with saline , MA , respectively , with a gradually increasing dose from 15 to 40 mg/kg for 12 weeks ( 5 days/week ) . Con A- and LPS-induced mitogenesis of splenocytes , cytokine production by splenocytes culture and lipid peroxides in the liver were measured . Heart tissue histopathology was analyzed in all the groups with murine cytomegalovirus ( CMV ) superinfection . Our data showed that MA treatment significantly decreased production of P60568 and interferon gamma ( P01579 ) in uninfected mice but did not further suppress the reduced Th1 cytokines in retrovirus-infected mice . There were no significant effects on cytokines P05112 and P05231 . However , tumor necrosis factor ( P01375 ) was significantly increased in both uninfected and infected mice due to MA treatment . Lipid peroxides in liver were significantly increased both in uninfected and retrovirus-infected mice due to MA exposure . DB00163 levels in liver were significantly decreased in uninfected mice due to MA treatment . CMV superinfection greatly increased the cardiac lesions in retrovirus-infected mice while no significant histopathology changes were detected due to MA treatment . Our data suggest that MA has immunomodulation activity , suppressing Th1 cytokine production and enhancing some Th2 cytokine secretion , as well as increasing lipid peroxides in uninfected mice . The interaction between LP-BM5 and MA remains unclear . 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 . DB08932 ( Opsumit ) for the treatment of pulmonary arterial hypertension . The endothelin pathway is a key pathway for the pathogenesis of pulmonary arterial hypertension ( PAH ) . Antagonism of this pathway is recommended as initial therapy in low-risk patient with PAH to inhibit fibrosis , cell proliferation , and inflammation caused by endothelin . Prior to October 2013 , ambrisentan , a selective P25101 receptor antagonist and DB00559 , a dual P25101 /ETB antagonist , were the only currently available agents for PAH targeting the endothelin pathway . Based on the results of the SERAPHIN trial , macitentan ( brand name Opsumit® ) , a new P25101 /ETB antagonist , has been US FDA approved to delay disease progression and reduce hospitalizations for PAH . SERAPHIN is the first ERA trial to use an event-driven strategy with a composite primary end point of morbidity or mortality . Previous trials have focused on short-term outcomes , such as improved 6-min walk distance and WHO functional class . Regulation and expression of endothelin-1 ( ET-1 ) and ET-receptors in rat epithelial cells of renal and intestinal origin . The hormone endothelin-1 ( ET-1 ) is involved in many functions of the kidney and intestine . In addition to its vasoactive and proliferative effects , ET-1 is involved in the maintenance of water and salt balance , and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs . To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro , we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal ( IEC-6 ) and renal ( Q7Z2Y5 -52E and Q9BZR9 ) origin . Immunocytochemistry showed that all three cell lines express ET-1 and the P25101 and P24530 receptor . ET-1 was expressed intracellularly , and also the P25101 receptor showed a punctate intracellular staining pattern . The P24530 receptor was localized in the membrane , which was confirmed by Western blot analysis . Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines , interleukin-1beta ( IL-1beta ) and tumor necrosis factor-alpha ( TNFalpha ) , induced ET-1 mRNA expression and excretion , while P60568 was ineffective . In Q7Z2Y5 -52E cells , IL-1beta and TNFalpha induced ET-1 excretion as well . In Q9BZR9 cells , adequate measurement of cytokine effects on ET-1 excretion was not possible , since ET-1 excretion under non-stimulated conditions was around the lowest level of detection . In conclusion , we showed ET-1 and ET-receptor expression , and inducibility of ET-1 by cytokines in IEC-6 , Q7Z2Y5 -52E , and Q9BZR9 cells . These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells . 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 . 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 . Translational research with pemetrexed in breast cancer . DB00642 ( Alimta ) is a novel folate antimetabolite that primarily inhibits the enzymes thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyl transferase ( GARFT ) , all of which are involved in pyrimidine and purine synthesis . In a phase II trial of patients with DB00279 /4 , N0-2 breast cancer , expression of thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , glycinamide ribonucleotide formyltransferase ( GARFT ) , p53 , and c-erb-B2 ( at the mRNA or protein level ) was examined in tumor biopsy specimens before and 24 hours after the first dose of pemetrexed and after three cycles of single-agent treatment to establish correlations of biomarker levels and changes with clinical outcome and toxicity . Although final data are not available , initial indications are that clinical response may correlate with decreased or low TS expression . The results obtained from clinical data are supported by laboratory results in three cell lines ( MDA-231 , MCF-7 , and ZR-75 ) . These results suggest that in vitro transcript profiling to identify which genes are important predictors of successful cytotoxic chemotherapy , followed by a focused clinical trial to confirm the in vitro results , may be the best approach for translational research . Cytological properties of stromal cells derived from giant cell tumor of bone ( GCTSC ) which can induce osteoclast formation of human blood monocytes without cell to cell contact . When human blood monocytes were cocultured with stromal cells derived from human giant cell tumor of bone ( GCTSC ) and a Millipore filter ( 0.4 microm ) was interposed between monocytes and GCTSC , multinucleated giant cell formation of monocytes was induced . The multinucleated giant cells have characters as osteoclast-like cells , indicating that a soluble osteoclast-inducing factor(s) is secreted from GCTSC expressing Q9Y6Q6 , O14788 / O14788 / O14788 and P78536 mRNA . Furthermore , O00300 / O00300 inhibited GCTSC-induced osteoclastogenesis , showing that the Q9Y6Q6 - O14788 system is involved in GCTSC-induced osteoclastogenesis and that soluble form of O14788 / O14788 induces osteoclasts from monocytes . GCTSC expressed the cytokine mRNAs such as P09603 , GM- P04141 , P08700 , P05112 , P05231 , and P01579 mRNAs . None of IL-1ralpha , IL-1alpha , IL-1beta , P60568 , P05112 , P22301 , Q14116 , P01375 , G- P04141 and P01579 could be detected in all culture media . A significant amount of P05231 could be detected in the culture media of all GCTSC . P10145 was found in the culture media of two GCTSC and two osteosarcoma-derived cells . P09603 was detected in all culture media . GCTSC express P41180 , and stimulation of GCTSC with either extracellular Ca(2+) or neomycin , agonist of P41180 , augmented the expression of O14788 . Some lines of GCTSC expressed alkaline phosphatase , osteocalcin and Cbfa1 , suggesting that GCTSC are intimately related to osteoblastic lineage . 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 . Phase II study of erlotinib ( DB00530 ) in patients with metastatic colorectal cancer . Erlotinib ( Tarceva , DB00530 ) , a potent epidermal growth factor receptor tyrosine kinase inhibitor ( P00533 ) , was evaluated in a phase II study to assess its activity in patients with metastatic colorectal cancer . In all , 38 patients with metastatic colorectal cancer were treated with erlotinib at a continuous daily oral dose of 150 mg . Radiological evaluation was carried out every 8 weeks and tumour biopsies were performed before treatment and on day 8 . Of 31 evaluable patients , 19 ( 61 % ) had progressive disease and 12 ( 39 % ) had stable disease ( s.d. ) . The median time to progression for those patients having s.d. was 123 days ( range 108-329 days ) . The most common adverse events were rash in 34 patients and diarrhoea in 23 patients . Correlative studies were conducted to investigate the effect of erlotinib on downstream signalling . Tumour tissue correlations were based on usable tissue from eight match paired tumour samples pre- and on therapy , and showed a statistically significant decrease in the median intensity of both pEGFR ( P=0.008 ) and phospho-extracellular signal-regulated kinase ( P29323 ) ( P=0.008 ) a week after commencement of treatment . No other statistically significant change in tumour markers was observed . Erlotinib was well tolerated with the most common toxicities being rash and diarrhoea . More than one-third of evaluable patients had s.d. for a minimum of 8 weeks . Correlative studies showed a reduction in phosphorylated P00533 and P29323 in tumour tissue post-treatment . 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 .

MH_train_1591

MH_train_1591

MH_train_1591

interacts_with DB01211?

multiple_choice

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 . DB06186 pharmacotherapy in patients with metastatic melanoma . Immune augmentation with ipilimumab , an anti- P16410 monoclonal antibody , has joined the ranks of approved immunologic agents for the treatment of metastatic melanoma . Phase III studies of ipilimumab in metastatic melanoma have demonstrated an overall survival advantage as compared to other approved and investigational therapies . However , the adverse effects associated with this medication are unique and often require management with steroids or other immunosuppressants . In addition , the time to response differs with ipilimumab as compared to traditional chemotherapy , and alternative means of assessment of response have been proposed . In this review , we will summarize the basic science of this treatment , its preclinical evaluation , and the clinical trials leading to its approval . We will also discuss the details regarding its use , assessment of response to this drug and other immune-related therapies , and further directions for investigation . Characterization of a ( BALB/c x C57BL/6 ) F1 T cell hybridoma with double specificity : recognition of antigen in context of I-Ad and autoreactivity to I-Ab . An antigen-specific , P60568 -producing , Lyt-1+2- , T cell hybridoma has been derived by the fusion of DB05299 ( KLH ) -primed ( BALB/c x C57BL/6 ) F1 lymph node with BW5147 thymoma cells . The hybridoma FN13-21 recognizes KLH in association with I-Ad of BALB/c parental antigen-presenting cells , and also responds to I-Ab of the other parent C57BL/6 spleen cells in the absence of antigen . The KLH-specific response of FN13-21 is blocked by monoclonal anti-I-Ad antibody , while the response to C57BL/6 spleen cells is blocked by anti-I-Ab antibody . The specificity of the autoreactivity is to an antigen encoded for in the I-Ab region , as shown by the pattern of stimulation obtained with spleen cells from H-2-recombinant mice . Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation : a novel role of the cysteinyl LT-1 receptor . Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues . Leukotrienes are potent inflammatory mediators , and similar to prostaglandins , are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues . Here we tested the hypothesis that blocking the negative regulation of leukotrienes , would lead to enhanced endochondral bone formation during fracture repair . A closed femoral fracture was created in mice . Animals were divided into three groups for treatment with either montelukast sodium , a cysteinyl leukotriene type 1 receptor antagonist ( trade name Singulair ) , zileuton , a P09917 enzyme inhibitor ( trade name DB00744 ) , or carrier alone . The fractures were analyzed using radiographs , quantitative gene expression , histology and histomorphometry , and immunohistochemistry . Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls . Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7 . Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation , and increased early bone formation markers when compared with controls . Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor , leading to increased chondrocyte proliferation at early time points . These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation , with important and previously unrecognized implications for both fracture repair , and in a broader context , systemic chondrocyte growth and differentiation . Characterization of the inhibitory effects of erythromycin and clarithromycin on the Q12809 potassium channel . Both erythromycin and clarithromycin have been reported to cause QT prolongation and the cardiac arrhythmia torsade de pointes in humans , however direct evidence documenting that these drugs produce this effect by blocking human cardiac ion channels is lacking . The goal of this study was to test the hypothesis that these macrolide antibiotics significantly block the delayed rectifier current ( IKr ) encoded by Q12809 ( the human ether-a-go-go-related gene ) at drug concentrations , temperature and ionic conditions mimicking those occurring in human subjects . DB01345 currents in P29320 293 cells stably transfected with Q12809 were recorded using a whole cell voltage clamp method . Exposure of cells to erythromycin reduced the Q12809 encoded potassium current in a concentration dependent manner with an IC50 of 38.9 +/- 1.2 microM and Hill Slope factor of 0.4 +/- 0.1 . DB01211 produced a similar concentration-dependent block with an IC50 of 45.7 +/- 1.1 microM and Hill Slope factor of 1.0 +/- 0.1 . Erythromycin ( 25-250 microM ) and clarithromycin ( 5 or 25 microM ) also produced a significant decrease in the integral of the current evoked by an action potential shaped voltage clamp protocol . The results of this study document that both erythromycin and clarithromycin significantly inhibit the Q12809 potassium current at clinically relevant concentrations . 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 . 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 . Expression of the human concentrative nucleotide transporter 1 ( O00337 ) gene correlates with clinical response in patients affected by Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) undergoing a combination treatment with 2-chloro-2'-deoxyadenosine ( DB00242 ) and DB00073 . PURPOSE : Resistance to nucleoside analogues agents is likely to be multifactorial and could involve a number of mechanisms affecting drug penetration , metabolism and targeting . In vitro studies of resistant human cell lines have confirmed that human concentrative nucleoside transporter 1 ( O00337 ) -deficient cells display resistance . EXPERIMENTAL DESIGN : We applied real-time PCR method to assess the mRNA expression of equilibrative and concentrative nucleoside transporter ( hENT1 , O00337 ) , deoxycytidine and deoxyguanosine kinase ( P27707 , Q16854 ) , 5'-nucleotidase ( 5'-NT ) , ribonucleotide reductase catalytic and regulatory ( P23921 , P31350 ) subunits in bone marrow cells from 32 patients with Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) who received 2CdA-based chemotherapy . Responses to chemotherapy , were then correlated to the expression of these markers . RESULTS : All 32 patients enrolled expressed lower levels of O00337 as compared to healthy donors . In univariate analysis , lower expression level of O00337 ( p=0.0021 ) and P31350 ( p=0.02 ) correlated with response to chemotherapy . In particular , patients with low levels of O00337 achieved inferior clinical response . No significant correlation between these genes expression and age , stage of disease was found . This study suggests that nucleotidase expression levels can be used to identify subgroups of WM and SLL patients who will likely respond differently to a 2CdA-based therapy . Additive role of tiotropium in severe asthmatics and Arg16Gly in P07550 as a potential marker to predict response . BACKGROUND : Recent findings have raised new interests about the use of anticholinergics , especially tiotropium , for the treatment of asthma . This study was performed to determine whether an additional improvement in lung function is obtained when tiotropium is administrated in addition to conventional therapies in severe asthmatics , and to identify factors capable of predicting the response to tiotropium , using a pharmacogenetic approach . METHODS : A total of 138 severe asthmatics on conventional medications and with decreased lung function were randomly recruited . DB01409 18 microg was added once a day and lung functions were measured every 4 weeks . Responders were defined as those with an improvement of > or = 15 % ( or 200 ml ) in the forced expiratory volume in 1 s ( FEV1 ) that was maintained for at least 8 successive weeks . Eleven single nucleotide polymorphisms ( SNPs ) in P11229 -3 ( coding muscarinic receptors one to three ) which were identified by re-sequencing , and Arg16Gly and Gln27Glu in P07550 ( coding beta(2) adrenoreceptor ) were scored in 80 of the 138 asthmatics . RESULTS : Forty-six of the 138 asthmatics ( 33.3 % ) responded to tiotropium treatment . Logistic regression analyses ( controlled for age , gender , and smoking status ) showed that Arg16Gly in P07550 [ P = 0.003 , OR ( 95 % CI ) = 0.21 ( 0.07-0.59 ) in a minor allele-dominant model ] was significantly associated with response to tiotropium . CONCLUSIONS : As many as 30 % of severe asthmatics on conventional medications with reduced lung function were found to respond to adjuvant tiotropium . The presence of Arg16Gly in P07550 may predict response to tiotropium . Farnesoid X receptor agonist for the treatment of liver and metabolic disorders : focus on 6-ethyl- DB06777 . 6-ethyl-chedeoxycholic acid ( 6E- DB06777 ) is a farnesoid X receptor ( Q96RI1 ) ligand endowed with agonistic activity under development for treatment of cholestatic liver diseases including primary biliary cirrhosis ( P10515 ) and liver-related metabolic disorders including non-alcoholic fatty liver disease ( NAFLD ) and non-alcoholic steatohepatitis ( NASH ) . Q96RI1 is a bile sensor that acts in coordination with other nuclear receptors to regulate essential steps of bile acid uptake , metabolism and excretion . 6E- DB06777 has been investigated in preclinical models of cholestasis , liver fibrosis and diet-induced atherosclerosis . In a phase II clinical trial in patients with P10515 , 6E- DB06777 met the primary endpoint of a reduction in alkaline phosphatase levels but safety data indicated that the drug exacerbated pruritus , one of the main symptoms of P10515 , suggesting that 6E- DB06777 or Q96RI1 are mediators of pruritus in humans . Treatment of patients with diabetes and liver steatosis resulted in amelioration of insulin sensitivity despite a reduction a slight reduction in HDL and increased levels of LDL were observed . These side effects on bile acids and lipid metabolism were all predicted by pre-clinical studies , suggesting that potent Q96RI1 ligands hold promise but potential side effects might limit their development . 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 . [ 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 . Mass spectrometric analysis of agonist effects on posttranslational modifications of the beta-2 adrenoceptor in mammalian cells . Posttranslational modifications ( PTMs ) of the beta-2 adrenoceptor ( P07550 ) play a fundamental role in receptor regulation by agonists . We have examined the effects of several agonists on net levels of P07550 palmitoylation and phosphorylation using epitope tagging in stably transfected human embryonal kidney ( P29320 ) 293 cells , immunoaffinity purification , and mass spectrometry combined with the method of stable isotope labeling by amino acids in cell culture ( SILAC ) . Palmitoylation of Cys341 was confirmed and did not change detectably after 30 min exposure of cells to saturating concentrations of dopamine , epinephrine , or isoproterenol . However , all of these agonists produced a marked increase in net phosphorylation . Phosphorylation of the third cytoplasmic loop was increased to a similar degree by all three agonists , whereas differences between agonists were observed in net phosphorylation of the carboxyl-terminal cytoplasmic domain ( isoproterenol approximately epinephrine >> dopamine ) . Interestingly , agonist-induced phosphorylation of the carboxyl-terminal cytoplasmic domain was observed exclusively in a proximal portion ( between residues 339-369 ) . None of the agonists produced detectable phosphorylation in a distal portion of the cytoplasmic tail , which contains all sites of agonist-induced phosphorylation identified previously by in vitro reconstitution . These results provide insight to agonist-dependent regulation of the P07550 in intact cells , suggest the existence of significant differences in regulatory phosphorylation events occurring between in vitro and in vivo conditions , and outline a general analytical approach to investigate regulated PTM of receptors in mammalian cells . Glucocorticoids regulate calcineurin-dependent trans-activating pathways for interleukin-2 gene transcription in human T lymphocytes . Glucocorticoids ( GC ) inhibit P60568 gene transcription by interfering with the binding of the nuclear factor activator protein-1 on the P60568 promoter . Calcineurin , a Ca2+/calmodulin-dependent protein phosphatase , is an essential component of the T cell antigen receptor signal transduction pathway leading to P60568 gene transcription . Therefore , we have asked whether this phosphatase may also be regulated by GC . Jurkat T cells were cotransfected with plasmids containing the intact P60568 promoter or its NF-AT and P14859 motifs , and a deletion mutant ( delta P62158 -AI ) of calcineurin known to have Ca(2+)-independent constitutive phosphatase activity . Cotransfection of P60568 promoter with delta P62158 -AI allowed the activation of P60568 promoter in the presence of phorbol ester alone . Under these conditions dexamethasone ( DB00514 ; 10(-6) M ) inhibited P60568 promoter activation by 50-60 % . The inhibitory effect of DB00514 was specific , as demonstrated by experiments using an unrelated promoter ( simian virus 40 ) and estradiol . Furthermore , it was completely reversed in the presence of excess amounts of the glucocorticoid antagonist RU 486 , which suggests that it is mediated through the glucocorticoid receptor . Overexpression of calcineurin via delta P62158 -AI in Jurkat cells decreased their apparent sensitivity to DB00514 ( approximately 5-fold increase in IC50 ) . Similar results were obtained with the NF-AT and P14859 constructs , which are also known to be activated by calcineurin . Thus , in addition to their known inhibitory effects on activator protein-1 , GC also inhibit calcineurin-dependent pathways for T cell activation . DB00952 mitigates CGRP1-associated motor neuron degeneration caused by an expanded polyglutamine repeat tract . Spinal and bulbar muscular atrophy ( SBMA ) is a motor neuron disease caused by the expansion of the CAG triplet repeat within the androgen receptor ( AR ) gene . Here , we demonstrated that pathogenic AR upregulates the gene encoding calcitonin gene-related peptide α ( CGRP1 ) . In neuronal cells , overexpression of CGRP1 induced cellular damage via the activation of the c-Jun N-terminal kinase ( JNK ) pathway , whereas pharmacological suppression of CGRP1 or JNK attenuated the neurotoxic effects of pathogenic AR . The depletion of CGRP1 inactivated JNK and suppressed neurodegeneration in a mouse model of SBMA . DB00952 , a serotonin 1B/1D ( 5-hydroxytryptamine 1B/1D , or P28222 /1D ) receptor agonist , decreased CGRP1 expression via the induction of dual-specificity protein phosphatase 1 ( P28562 ) , attenuated JNK activity and mitigated pathogenic AR-mediated neuronal damage in cellular and mouse SBMA models . These observations suggest that pharmacological activation of the P28222 /1D receptor may be used therapeutically to treat SBMA and other polyglutamine-related neurodegenerative diseases . Biliary cytokeratin expression but not CD56 ( N- P62158 ) expression aids in the differential diagnosis of non-neoplastic bile duct diseases . Non-neoplastic bile duct diseases include several entities with a variety of clinical and histopathologic features . In needle biopsies , however , these may overlap . Here , auxiliary diagnostic markers would be helpful . CD56 ( N- P62158 ) has been reported in bile duct development , liver regeneration , and different liver diseases . This study was performed to evaluate the diagnostic value of CD56 immunohistochemistry compared to biliary cytokeratins in the diagnosis of non-neoplastic biliary liver diseases in liver needle biopsies . Thirty-eight cases ( 10× PSC ; 10× P10515 ; 10× obstruction ; 8× drug-induced liver disease [ DILD ] ) were analyzed using antibodies against CD56/ P13591 , CK7 , and CK19 . Twenty-three of all cases ( 63.9 % ) showed a positive CD56 reaction ( PSC 6/10 ; P10515 9/10 ; obstruction 5/10 ; DILD 3/8 ) with no statistical significance between the groups . Biliary cytokeratins visualized the bile ducts in all cases . CK7 highlighted cholangiolar metaplasia in seven cases ( 3× PSC ; 1× P10515 ; 3× DILD ) . CD56 can not be used as a supplementary tool in the differential diagnosis of non-neoplastic biliary diseases . CK7 should be included in the routine assessment of liver biopsies in these settings . Further research is needed to find better targets for immunohistochemical determination of the etiology of bile duct damage . P13569 modulates neurosecretory function in pulmonary neuroendocrine cell-related tumor cell line models . The pulmonary neuroendocrine cell ( PNEC ) system consists of solitary cells and distinctive cell clusters termed neuroepithelial bodies ( P20929 ) localized in the airway epithelium . PNEC/ P20929 express a variety of bioactive substances , including amine ( serotonin , 5HT ) and neuropeptides . We have previously shown that P20929 cells are O(2) sensors expressing nicotinamide adenine diphosphate oxidase complex and O(2) sensitive K(+) channel . Recently , we demonstrated expression of functional cystic fibrosis transmembrane conductance regulator ( P13569 ) and Cl(-) conductances in P20929 cells of rabbit neonatal lung . Because PNEC/ P20929 are sparsely distributed and difficult to study in native lung , we investigated small-cell lung carcinoma ( SCLC ) and carcinoid tumor cell lines ( tumor counterparts of normal PNEC/ P20929 ) as models for PNEC/ P20929 . SCLC ( H146 , H345 ) and carcinoid ( H727 ) cell lines express neuroendocrine cell markers , including chromogranin A , neural cell adhesion molecule ( N- P62158 ) , 5HT , and tryptophan hydroxylase . We report that H146 , H345 , and H727 express P13569 messenger RNA ( reverse transcription polymerase chain reaction ) and protein ( immunoblotting ) and possess functional P13569 Cl(-) conductance , demonstrated by an iodide efflux assay inhibitable by transfection with antisense P13569 . Using an immunoassay to quantitate 5HT secretion , we also show that downregulation of P13569 abolishes hypoxia-induced 5HT release , and reduces secretory response to high potassium . Our findings suggest that P13569 may modulate neurosecretory activity of PNEC/ P20929 possessing O(2) sensor function . We propose that these tumor cell lines may be useful models for investigating the role of P13569 in PNEC/ P20929 functions in health and disease . Intracellular capture of P33681 in antigen-presenting cells reduces costimulatory activity . P16410 gene constructs were designed to express P16410 exclusively in the endoplasmic reticulum ( ER ) . Four different P16410 gene constructs were transfected into P29320 293 ( human embryonic kidney ) and A20 ( Balb/c mouse B lymphoma ) cells . All constructs contained an ER retention signal and coded for P16410 expression in the ER . One of the constructs , which contained the membrane part of P16410 , coded for an expression both on the cell surface and in the ER . Three of the expressed P16410 types ( including the ER-membrane-expressed form ) caused a reduced surface expression of P33681 in the A20 cells . Only constructs which allow dimerization of P16410 showed this effect . It is assumed that intracellular P16410 bound P33681 and inhibited therefore the transport of P33681 to the surface . The binding obviously caused also an enhanced degradation of the complexes because both proteins showed a low concentration in the transfected cell lines . P16410 -transfected and P33681 -reduced A20 cells showed a diminished costimulating activity upon T cells . This was demonstrated by a reduced proliferation of T cells from ovalbumin-immunized Balb/c mice , incubated with ovalbumin peptide-primed P16410 -transfected A20 cells . 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 . Coaggregation of RNA-binding proteins in a model of Q13148 proteinopathy with selective RGG motif methylation and a role for P23921 ubiquitination . Q13148 ( Q13148 ) is a major component within ubiquitin-positive inclusions of a number of neurodegenerative diseases that increasingly are considered as Q13148 proteinopathies . Identities of other inclusion proteins associated with Q13148 aggregation remain poorly defined . In this study , we identify and quantitate 35 co-aggregating proteins in the detergent-resistant fraction of P29320 -293 cells in which Q13148 or a particularly aggregate prone variant , TDP-S6 , were enriched following overexpression , using stable isotope-labeled ( SILAC ) internal standards and liquid chromatography coupled to tandem mass spectrometry ( LC-MS/MS ) . We also searched for differential post-translational modification ( PTM ) sites of ubiquitination . Four sites of ubiquitin conjugation to Q13148 or TDP-S6 were confirmed by dialkylated Q86UG4 - Q13148 external reference peptides , occurring on or near RNA binding motif ( RRM ) 1 . RRM-containing proteins co-enriched in cytoplasmic granular structures in P29320 -293 cells and primary motor neurons with insoluble TDP-S6 , including cytoplasmic stress granule associated proteins Q13283 , P11940 , and eIF4A1 . Proteomic evidence for Q13148 co-aggregation with paraspeckle markers Q96PK6 , PSF and NonO was also validated by western blot and by immunocytochemistry in P29320 -293 cells . An increase in peptides from methylated arginine-glycine-glycine ( RGG ) RNA-binding motifs of P35637 / P35637 and hnRNPs was found in the detergent-insoluble fraction of TDP-overexpressing cells . Finally , Q13148 and TDP-S6 detergent-insoluble species were reduced by mutagenesis of the identified ubiquitination sites , even following oxidative or proteolytic stress . Together , these findings define some of the aggregation partners of Q13148 , and suggest that Q13148 ubiquitination influences Q13148 oligomerization . DB01211 substantially increases steady-state DB00559 exposure in healthy volunteers . AIMS : The aim of this study was to assess the effect of the cytochrome P450 ( CYP ) 3A4 and organic anion-transporting polypeptide ( P46721 ) 1B1 inhibitor clarithromycin on the pharmacokinetics of DB00559 . We also aimed to evaluate the impact of P11712 and Q9Y6L6 ( encoding for Q9Y6L6 ) genotypes and their combination . METHODS : We assessed the effect of the P46721 and CYP3A inhibitor clarithromycin on DB00559 pharmacokinetics at steady state and concurrently quantified changes of CYP3A activity using midazolam as a probe drug . Sixteen healthy volunteers received therapeutic doses of DB00559 ( 125 mg twice daily ) for 14 days and clarithromycin ( 500 mg twice daily ) concomitantly for the last 4 days , and DB00559 pharmacokinetics was assessed on days 1 , 10 and 14 . RESULTS : DB01211 significantly increased DB00559 area under the plasma concentration-time curve of the dosing interval 3.7-fold and peak concentration 3.8-fold in all participants irrespective of the genotype . DB01211 also reduced CYP3A activity ( midazolam clearance ) in all participants ; however , these changes were not correlated to the changes of DB00559 clearance . CONCLUSIONS : DB01211 substantially increases the exposure to DB00559 , suggesting that dose reductions may be necessary .

MH_train_1592

MH_train_1592

MH_train_1592

interacts_with DB00864?

multiple_choice

Signaling by proinflammatory cytokines : oligomerization of TRAF2 and Q9Y4K3 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain . Interleukin-1 ( IL-1 ) and tumor necrosis factor ( P01375 ) stimulate transcription factors AP-1 and NF-kappaB through activation of the Q96HU1 kinases JNK and p38 and the O15111 ( IKK ) , respectively . The P01375 and IL-1 signals are transduced through TRAF2 and Q9Y4K3 , respectively . Overexpressed TRAF2 or Q9Y4K3 activate JNK , p38 , or IKK in the absence of extracellular stimulation . By replacing the carboxy-terminal TRAF domain of TRAF2 and Q9Y4K3 with repeats of the immunophilin P62942 , we demonstrate that their effector domains are composed of their amino-terminal Zn and RING fingers . Oligomerization of the TRAF2 effector domain results in specific binding to Q13233 , a protein kinase capable of JNK , p38 , and IKK activation , and induction of P01375 and IL-1 responsive genes . P01375 also enhances the binding of native TRAF2 to Q13233 and stimulates the kinase activity of the latter . Thus , P01375 and IL-1 signaling is based on oligomerization of TRAF2 and Q9Y4K3 leading to activation of effector kinases . Activin A augments DB00644 -mediated transcriptional activation of the mouse P30968 gene . The response of pituitary gonadotropes to DB00644 correlates directly with the concentration of DB00644 receptors ( GnRHRs ) on the cell surface , which is mediated in part at the level of GnRHR gene expression . We have previously localized DB00644 responsiveness in the mouse GnRHR ( mGnRHR ) gene promoter to two elements : activating protein-1 and sequence underlying responsiveness to DB00644 -1 . This study was designed to investigate potential synergy between DB00644 and activin A in transcriptional activation of the mGnRHR gene . In functional transfection studies using alphaT3-1 cells , DB00644 agonist stimulation of the mGnRHR gene promoter ( -765/+62 ) resulted in a 10.9-fold increase in activity , which was further increased 2-fold ( to 21.3-fold ) following activin pretreatment . Activin pretreatment alone had no effect . Deletion of region -387/-308 or mutation of a putative SMAD-binding element at -331/-324 ( 5'-GTCTAG[T]C-3' ) abrogated the augmented response to DB00644 in the presence of activin but not the response to DB00644 alone . Overexpression of Q15796 and P84022 along with Q13485 increased transcriptional activity of the mGnRHR gene , which was further increased by DB00644 agonist stimulation . These data demonstrate that activin augments DB00644 -mediated transcriptional activation of the mGnRHR gene and suggest that this effect may be mediated through SMAD transcription factors . The complex of FK506-binding protein 12 and FK506 inhibits calcineurin phosphatase activity and IgE activation-induced cytokine transcripts , but not exocytosis , in mouse mast cells . FK506 and cyclosporin A ( DB00091 ) are immunosuppressive agents that inhibit P60568 production by activated T cells , but only DB00091 inhibits IgE activation-induced cytokine transcripts in mouse P08700 -dependent , bone marrow-derived mast cells ( BMMC ) . We previously associated the resistance of BMMC to FK506 with a deficiency in the expression of FK506 binding protein ( FKBP ) 12 , a molecule that forms a complex with FK506 capable of inhibiting calcineurin phosphatase activity in vitro . In this report , we establish that P62942 mediates FK506 inhibition of both calcineurin phosphatase activity and IgE activation-induced cytokine transcripts in a Kirsten murine sarcoma virus-immortalized mast cell line that is P62942 deficient . Overexpression of P62942 by transfection enhanced the ability of FK506 to inhibit calcineurin phosphatase activity ( IC50 = 2 nM ) , compared with cells transfected with the expression vector alone ( IC50 > 30 nM ) . The IC50 value for FK506 inhibition of IgE activation-induced transcripts for P01375 decreased from 40 nM in vector control cells to 10 nM in P62942 transfectants . Similarly , the IC50 value for inhibition of P05231 transcripts decreased from > 1000 nM in vector control cells to 35 nM in P62942 transfectants . In contrast , activation-elicited release of the secretory granule mediator beta-hexosaminidase was only partially inhibited by FK506 at 1000 nM , regardless of the levels of P62942 expressed by the cells . Thus , P62942 is the dominant cytosolic protein that mediates FK506 inhibition of P01375 and P05231 transcripts . P12004 D3 is down-regulated by rapamycin in HER-2-overexpressing breast cancer cells . DB00877 and its analogues are being tested as new antitumor agents . DB00877 binds to P62942 and this complex inhibits the activity of P42345 /mammalian target of rapamycin , which leads to dephosphorylation of 4EBP1 and P08133 S6 kinase , resulting in blockade of translation initiation . We have found that P30533 inhibits the growth of HER-2-overexpressing breast cancer cells . The phosphorylation of mammalian target of rapamycin , P08133 S6 kinase , and 4EBP1 is inhibited by rapamycin and cells are arrested in the P55008 phase , as determined by growth assays , fluorescence-activated cell sorting analysis , and bromodeoxyuridine incorporation studies . DB00877 causes down-regulation of cyclin D3 protein , retinoblastoma hypophosphorylation , loss of cyclin-dependent kinase ( cdk ) 4 , cdk6 , and cdk2 activity . The half-life of cyclin D3 protein decreases after rapamycin treatment , but not its synthesis , whereas the synthesis or half-life of cyclin D1 protein is not affected by the drug . Additionally , rapamycin caused accumulation of ubiquitinated forms of cyclin D3 protein , proteasome inhibitors blocked the effect of rapamycin on cyclin D3 , and rapamycin stimulated the activity of the proteasome , showing that the effect of rapamycin on cyclin D3 is proteasome proteolysis dependent . This effect depends on the activity of HER-2 because Herceptin , a neutralizing antibody against HER-2 , is able to block both the induction of proteasome activity and the cyclin D3 down-regulation due to rapamycin . Furthermore , inhibition of HER-2 gene expression by using small interfering RNA blocked the rapamycin effects on cyclin D3 . These data indicate that rapamycin causes a P55008 arrest in HER-2-overexpressing breast cancer cells that is associated with a differential destabilization and subsequent down-regulation of cyclin D3 protein . DB06366 in the treatment of P04626 + breast cancer . DB06366 , a humanized monoclonal antibody and the first in the class of agents called the P04626 dimerization inhibitors , impairs the ability of P04626 to bind to other members of the HER family . It has a unique and complimentary mechanism of action compared with trastuzumab , and the combination has resulted in the enhanced blockade of the HER signaling pathway . When pertuzumab was used in combination with docetaxel and trastuzumab in the first-line treatment of metastatic P04626 (+) breast cancer , it led to an overall survival benefit . DB06366 has therefore been approved by the FDA and is currently used as a standard of care for this indication . It is also the first agent in oncology to receive accelerated FDA approval in the neoadjuvant setting . Randomized trials showed that the addition of pertuzumab to trastuzumab-based chemotherapy improves pathologic complete response rates in P04626 (+) early-stage breast cancer . A randomized phase III clinical trial with disease-free survival as the primary end point is evaluating the safety and efficacy of pertuzumab in the adjuvant setting . This article describes the preclinical data , synthesizes available data from phase I-III clinical trials of pertuzumab in early stage and metastatic settings , and puts them into perspective with current treatment recommendations and future research developments . Array-based comparative genomic hybridization identifies P11802 and Q08050 alterations as independent predictors of survival in malignant peripheral nerve sheath tumor . PURPOSE : Malignant peripheral nerve sheath tumors ( MPNST ) are highly aggressive sarcomas with variable patient survival and few known prognostically relevant genomic biomarkers . To identify survival-associated genomic biomarkers , we performed high-resolution array-based comparative genomic hybridization ( aCGH ) on a large set of MPNSTs . EXPERIMENTAL DESIGN : Candidate gene alterations identified by aCGH in 38 MPNSTs were validated at the DNA , RNA , and protein levels on these same tumors and an independent set of 87 MPNST specimens . RESULTS : aCGH revealed highly complex copy number alterations , including both previously reported and completely novel loci . Four regions of copy number gain were associated with poor patient survival . Candidate genes in these regions include P35711 ( 12p12.1 ) , P46087 and Q15773 ( 12p13.31 ) , Q08050 and P62942 ( 12p13.33 ) , and P11802 and Q12999 ( 12q14.1 ) . Alterations of these candidate genes and several others of interest ( P04626 , MYC and P04637 ) were confirmed by at least 1 complementary methodology , including DNA and mRNA quantitative real-time PCR , mRNA expression profiling , and tissue microarray-based fluorescence in situ hybridization and immunohistochemistry . Multivariate analysis showed that P11802 gain/amplification and increased Q08050 protein expression were the most significant independent predictors for poor survival in MPNST patients ( P < 0.05 ) . CONCLUSIONS : Our study provides new and independently confirmed candidate genes that could serve as genomic biomarkers for overall survival in MPNST patients . The preclinical development of regorafenib for the treatment of colorectal cancer . INTRODUCTION : The DB01367 -RAF-MEK- P29323 pathway is one of the best characterized kinase cascades . During the exploration of small molecules that inhibit P04049 kinase , regorafenib ( DB08896 ) was discovered as a multikinase inhibitor which demonstrated anti-cancer , anti-angiogenic , and apoptotic activities in metastatic colorectal cancer . This was not the first multikinase inhibitor discovered for the disease ; indeed , before regorafenib was approved by FDA as a multikinase inhibitor for metastatic colorectal cancer in 2012 , sorafenib ( BAY 43-9006 ) had already been developed to be the first in the world as a multikinase inhibitor for malignancy . Indeed , the only difference between the two compounds is fluorine bound to its proximal phenyl ring although the end result is a considerably different profile , both as a kinase inhibitor as well as in its clinical application . AREAS COVERED : In this drug discovery case history , the authors review the design , discovery , and development of both regorafenib and sorafenib from back in the 1990s . Furthermore , the authors highlight the drug 's anti-cancer and anti-angiogenic properties as well as its efficacy , safety pharmacology and toxicology based on FDA documents . EXPERT OPINION : In order to better predict the efficacy of kinase inhibitors and to utilize them more efficiently , our understanding of drug discovery , the approaches for kinase profiling , and technologies needed for their development are paramount . Indeed , the authors believe that the field should better explore the use of predictive biomarkers that might be able to better assess these therapeutics . Pharmaceutical scientists must also consider the cost effectiveness of the targeted agents developed as a number of the drugs developed are very expensive . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Phosphodiesterase 4 inhibitor cilomilast inhibits fibroblast-mediated collagen gel degradation induced by tumor necrosis factor-alpha and neutrophil elastase . Tissue destruction , resulting in emphysema , can be a consequence of several pathologic processes . The current study evaluated the effects of the phosphodiesterase (PDE)4 inhibitor , cilomilast , and other PDE inhibitors on the ability of fibroblasts to degrade extracellular matrix . Using the three-dimensional collagen gel culture system , fibroblasts ( HFL-1 ) were cultured with tumor necrosis factor ( P01375 ) -alpha , known to induce matrix metalloproteinase ( MMP ) release , and/or neutrophil elastase ( NE ) , which can induce MMP activation . On Day 4 , gels containing P01375 and NE were significantly degraded ( 20.8 +/- 2.9 % of original collagen content ) . DB03849 ( 10 micro M ) inhibited this degradation ( 84.4 +/- 8.4 % ) . DB01427 , a PDE3 inhibitor , and zaprinast , a O76074 inhibitor , had no effect . Gelatin zymography and immunoblotting revealed that fibroblasts cultured with P01375 released increased amounts of latent P03956 and -9 . The addition of NE resulted in the conversion of P03956 and -9 to their active forms , indicative of collagen degradation . DB03849 inhibited the release of P03956 and -9 , as well as conversion of P03956 to its active form . Using real-time PCR analysis , cilomilast 's effect on P03956 release was not associated with the proteinase 's mRNA expression , suggesting that the inhibition of release is regulated at the post-transcriptional level . These results suggest that cilomilast may be a potentially effective therapeutic agent in diseases characterized by excessive tissue destruction , such as emphysema . P43119 in tumor endothelial cells promotes angiogenesis in an autocrine manner . Molecules highly expressed in tumor endothelial cells ( TEC ) are important for specific targeting of these cells . Previously , using DNA microarray analysis , we found that the prostacyclin receptor ( IP receptor ) gene was upregulated in TEC compared with normal endothelial cells ( NEC ) . Although prostacyclin is implicated in re-endothelialization and angiogenesis , its role remains largely unknown in TEC . Moreover , the effect of the IP receptor on TEC has not been reported . In the present study we investigated the function of the IP receptor in TEC . The TEC were isolated from two types of human tumor xenografts in nude mice , while NEC were isolated from normal counterparts . DB01240 secretion levels in TEC were significantly higher than those in NEC , as shown using ELISA . Real-time RT-PCR showed that the IP receptor was upregulated in TEC compared with NEC . Furthermore , migration and tube formation of TEC were suppressed by the IP receptor antagonist RO1138452 . Immunohistostaining showed that the IP receptor was specifically expressed in blood vessels of renal cell carcinoma specimens , but not in glomerular vessels of normal renal tissue . These findings suggest that the IP receptor is a TEC-specific marker and might be a useful therapeutic target . Ablation of microvessels in vivo upon dimerization of iCaspase-9 . Anti-angiogenic therapies based on targeted disruption of the tumor microvascular network have been proposed for cancer treatment . Inhibitors of the endothelial cell pro-survival pathway mediated by P15692 were shown to activate caspases and cause microvascular regression , but the efficacy of this strategy can be hindered by the engagement of redundant survival pathways . Alternatively , if direct activation of an apical pro-apoptotic caspase is sufficient to disrupt microvessels in vivo , such a strategy could potentially override upstream endothelial cell survival inputs and disrupt tumor neovascular networks . Here , we fused caspase-9 to a mutated P62942 domain to express an inducible caspase-9 molecule ( iCaspase-9 ) that can be activated by a cell-permeable dimerizer drug , and transduced this construct into primary endothelial cells . We found that drug-induced dimerization of iCaspase-9 is sufficient to activate endogenous caspase-3 and trigger apoptosis even when endothelial cells are treated with the pro-survival factors P15692 or P09038 . A single intraperitoneal injection of the dimerizer drug induced apoptosis of endothelial cells expressing iCaspase-9 and elimination of human microvessels engineered in immunodeficient mice . These results demonstrate that the activation of iCaspase-9 disrupts microvessels in vivo , and suggest a novel anti-angiogenic strategy based on the expression and controlled activation of an inducible death gene in neovascular endothelial cells . P43119 suppresses cardiac fibrosis : role of CREB phosphorylation . Cardiac fibrosis is a consequence of many cardiovascular diseases and contributes to impaired ventricular function . Activation of the prostacyclin receptor ( IP ) protects against cardiac fibrosis , but the molecular mechanisms are not totally understood . Using mouse cardiac fibroblasts , we found that IP activation with cicaprost suppressed expression of collagen I and other target genes of transforming growth factor-beta . This effect of cicaprost was unlikely to be mediated by inhibition of the Q15796 /3 or mitogen-activated protein kinase ( MAPK ) activities , but was associated with DB02527 elevation and phosphorylation of the transcription factor DB02527 response element binding protein ( CREB ) . Expression of a non-phosphorylated CREB mutant suppressed the inhibitory effect of cicaprost . It appears that phosphorylated CREB binds to and sequestrates the transcription coactivator CBP/p300 from binding to Smad . Inhibition of the intrinsic histone acetyl-transferase activity of CBP/p300 with garcinol significantly suppressed collagen I expression in fibroblasts . Using apolipoprotein E and IP double knockout mouse , we demonstrated that endogenous prostacyclin/IP signaling had an inhibitory effect on angiotensin II-induced cardiac fibrosis under hypercholesterolemic conditions . Taken together , our results suggest that the prostacyclin/IP pathway suppresses cardiac fibrosis , at least partly , by inducing CREB phosphorylation . Effects of gonadoliberin analogue triptorelin on the pituitary-testicular complex in neonatal rats . DB06825 , a synthetic analogue of neurohormone gonadoliberin ( gonadotropin-releasing hormone , DB00644 ) administered daily to rats on postnatal days 5-7 suppressed the expression of P30968 in the pituitary gland , but did not change functioning of the pituitary-testicular complex . Administration of triptorelin on postnatal days 12-14 ( i.e. during the formation of pulsatile pattern of DB00644 secretion and increasing levels of its mRNA receptor in the pituitary gland ) had no effect on receptor expression , but increased the levels of luteinizing hormone mRNA in the pituitary gland and the weight of testes . At that time , blood levels of testosterone were lowered , which indicated disturbed pulsatile pattern of DB00644 secretion . Determination of cobimetinib in human plasma using protein precipitation extraction and high-performance liquid chromatography coupled to mass spectrometry . Inhibition of Q96HU1 / P29323 kinase ( MEK ) is a promising strategy to control the growth of tumors that are dependent on aberrant signaling in the MEK pathway . DB05239 ( P16260 -0973 ) ( S ) -[3,4-Difluoro-2-(2-fluoro-4-iodo-phenylamino)-phenyl]- ( ( S ) -3-hydroxy-3-piperidin-2-yl-azetidin-1-yl ) -methanone ) inhibits proliferation of a variety of human tumor cell lines by inhibiting Q02750 and P36507 . A specific high performance liquid chromatography-mass spectrometric assay was developed and validated for the determination of cobimetinib in human plasma . The overall mean recovery using protein precipitation extraction with acetonitrile was found to be 54.1 % . The calibration curve was ranged from 0.20 to 100ng/mL . The LLOQ was sensitive enough to detect terminal phase concentrations of the drug . The intra- and inter-assay precision ( % CV ) was within 10.3 % and 9.5 % for cobimetinib . The assay accuracy ( % RE ) was within ±13.7 % of the nominal concentration values for cobimetinib with the normal analytical QCs . The developed assay was successfully used to analyze the human plasma samples ( for pharmacokinetic analysis ) from clinical trials . Endothelial cell transforming growth factor-β receptor activation causes tacrolimus-induced renal arteriolar hyalinosis . Arteriolar hyalinosis is a common histological finding in renal transplant recipients treated with the calcineurin inhibitor tacrolimus ; however , the pathophysiologic mechanisms remain unknown . In addition to increasing transforming growth factor ( TGF ) -β levels , tacrolimus inhibits calcineurin by binding to FK506-binding protein 12 ( P62942 ) . P62942 alone also inhibits TGF-β receptor activation . Here we tested whether tacrolimus binding to P62942 removes an inhibition of the TGF-β receptor , allowing ligand binding , ultimately leading to receptor activation and arteriolar hyalinosis . We found that specific deletion of P62942 from endothelial cells was sufficient to activate endothelial TGF-β receptors and induce renal arteriolar hyalinosis in these knockout mice , similar to that induced by tacrolimus . DB00864 -treated and knockout mice exhibited significantly increased levels of aortic TGF-β receptor activation as evidenced by Q15796 /3 phosphorylation , along with increased collagen and fibronectin expression compared to controls . Treatment of isolated mouse aortas with tacrolimus increased TGF-β receptor activation and collagen and fibronectin expression . These effects were independent of calcineurin , absent in endothelial denuded aortic rings , and could be prevented by the small molecule TGF-β receptor inhibitor SB-505124 . Thus , endothelial cell TGF-β receptor activation is sufficient to cause vascular remodeling and renal arteriolar hyalinosis . 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 . DB09079 , a triple angiokinase inhibitor , enhances cytotoxic therapy response in pancreatic cancer . Angiogenesis remains a sensible target for pancreatic ductal adenocarcinoma ( PDAC ) therapy . P15692 , PDGF , FGF and their receptors are expressed at high levels and correlate with poor prognosis in human PDAC . DB09079 is a triple angiokinase inhibitor that targets P17948 /2/3 , P11362 /2/3 and PDGFRα/β signaling . We investigated the antitumor activity of nintedanib alone or in combination with the cytotoxic agent gemcitabine in experimental PDAC . DB09079 inhibited proliferation of cells from multiple lineages found in PDAC , with gemcitabine enhancing inhibitory effects . DB09079 blocked PI3K/MAPK activity and induced apoptosis in vitro and in vivo . In a heterotopic model , net local tumor growth compared to controls ( 100 % ) was 60.8 ± 10.5 % in the gemcitabine group , -2.1 ± 9.9 % after nintedanib therapy and -12.4 ± 16 % after gemcitabine plus nintedanib therapy . Effects of therapy on intratumoral proliferation , microvessel density and apoptosis corresponded with tumor growth inhibition data . In a PDAC survival model , median animal survival after gemcitabine , nintedanib and gemcitabine plus nintedanib was 25 , 31 and 38 days , respectively , compared to 16 days in controls . The strong antitumor activity of nintedanib in experimental PDAC supports the potential of nintedanib-controlled mechanisms as targets for improved clinical PDAC therapy . 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 . P62942 , the 12-kDa FK506-binding protein , is a physiologic regulator of the cell cycle . P62942 , the 12-kDa FK506-binding protein , is a ubiquitous abundant protein that acts as a receptor for the immunosuppressant drug FK506 , binds tightly to intracellular calcium release channels and to the transforming growth factor beta ( TGF-beta ) type I receptor . We now demonstrate that cells from P62942 -deficient ( P62942 (-/-) ) mice manifest cell cycle arrest in G(1) phase and that these cells can be rescued by P62942 transfection . This arrest is mediated by marked augmentation of P38936 ( P38936 /CIP1) levels , which can not be further augmented by TGF-beta1 . The P38936 up-regulation and cell cycle arrest derive from the overactivity of TGF-beta receptor signaling , which is normally inhibited by P62942 . Cell cycle arrest is prevented by transfection with a dominant-negative TGF-beta receptor construct . TGF-beta receptor signaling to gene expression can be mediated by SMAD , p38 , and P29323 / Q96HU1 kinase ( extracellular signal-regulated kinase/mitogen-activated protein kinase ) pathways . SMAD signaling is down-regulated in P62942 (-/-) cells . Inhibition of P29323 / Q96HU1 kinase fails to affect P38936 up-regulation . By contrast , activated phosphorylated p38 is markedly augmented in P62942 (-/-) cells and the P38936 up-regulation is prevented by an inhibitor of p38 . Thus , P62942 is a physiologic regulator of cell cycle acting by normally down-regulating TGF-beta receptor signaling .

MH_train_1593

MH_train_1593

MH_train_1593

interacts_with DB06684?

multiple_choice

P07237 -mediated ER retention and proteasomal degradation of procollagen I in corneal endothelial cells . Procollagen I in corneal endothelial cells ( CECs ) is intracellularly degraded immediately after its synthesis . In this study , we investigated the mechanism of intracellular degradation of procollagen I by determining the role of protein disulfide isomerase ( P07237 ) in endoplasmic reticulum ( ER ) retention and further determined the degradation pathway of procollagen I in CECs . When association of P07237 to monomeric proalpha chains or the trimeric procollagen I carboxyl propeptides ( PICPs ) was analyzed , immune complex precipitated with anti-PICP antibody contained more P07237 than that precipitated with antibodies to monomeric chains . PICPs were completely colocalized with P07237 . When CECs were transfected with P07237 vector , procollagen I and the recombinant P07237 were colocalized in the ER , whereas CECs transfected with P07237 minus KDEL ( the ER retrieval sequence ) vector demonstrated that the two proteins were localized in the Golgi and were subsequently secreted into the medium . DB03615 ( an inhibitor of the chaperone activity of P07237 ) blocked colocalization of P07237 and procollagen I . Cells treated with chloroquine ( lysosome inhibitor ) did not alter the subcellular localization of procollagen I , because the inhibitor failed to induce the accumulation of procollagen I at Golgi . On the other hand , procollagen I was colocalized with ubiquitin in the cytoplasm , and proteasomal inhibitors further facilitated the colocalization of the two proteins and accumulation of ubiquitinated procollagen I ladders . These results suggest that association of P07237 with procollagen I , whether monomeric or trimeric , leads to ER retention of procollagen I before intracellular degradation via the ubiquitin-proteasome pathway . Osteoclast progenitors from cats with and without tooth resorption respond differently to 1,25-dihydroxyvitamin D and interleukin-6 . Both vitamin D and inflammatory cytokines can stimulate osteoclast formation and activity . We studied the effect of DB00136 ( 1,25(OH)(2)D ) , and interleukin-6 ( P05231 ) , on the formation and activity of feline osteoclasts , using peripheral blood mononuclear cells ( PBMCs ) from cats with and without tooth resorption ( TR(+) and TR(-) ) as a source of osteoclast precursors . The formation of osteoclast-like cells ( defined as multinucleated , tartrate-resistant acid phosphatase-positive cells ) was assessed at 7 and 14 days . In the presence of P09603 and O14788 , with and without P05231 , more osteoclasts were formed from TR(-) PBMCs than from TR(+) PBMCs on plastic . More osteoclasts were formed from TR(+) PBMCs on bone slices in the presence of P09603 / O14788 with 1,25(OH)(2)D . This opposite effect may be due to a higher expression of the vitamin D receptor ( P11473 ) in TR(+) osteoclasts and precursors on bone . Formation of resorption pits was analyzed and confirmed with scanning electron microscopy . In conclusion , we propose that TR(+) PBMCs when cultured on bone are sensitive to 1,25(OH)(2)D , whereas the differentiation of TR(-) PMBCs on bone seem more sensitive to P05231 , suggesting that osteoclast precursors from cats with and without tooth resorption respond differently to osteoclast stimulating factors . Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO:fat:protein = 12:59:28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO:fat:protein = 56:24:20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n-6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4n-6 ) , while its biosynthetic metabolic intermediates were decreased . The n-6/n-3 and arachidonic/eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1n-7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 , P05231 , P10145 , P13500 , P16581 , I- P62158 , and P05121 . Increased 20:4n-6 and the ratios of 20:4n-6/20:5n-3 and n-6/n-3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet . 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 . 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 . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . 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 . Inhibition of corneal inflammation by the O00206 antagonist DB04933 tetrasodium ( E5564 ) . PURPOSE : To investigate the role of the O00206 /MD-2 antagonist eritoran tetrasodium in a murine model of contact lens-associated corneal infiltrates . METHODS : C57BL/6 mouse corneas were abraded and treated with eritoran tetrasodium or placebo , either before or after stimulation with either LPS , the O60603 ligand Pam(3) DB00151 , or antibiotic-killed Pseudomonas aeruginosa . A 2-mm punch from a silicon hydrogel contact lens was used to cover the corneal surface throughout the inhibition and stimulation period . Corneal infiltrates were detected by in vivo confocal microscopy and by immunohistochemistry for neutrophils . The effect of eritoran tetrasodium on stimulated human corneal epithelial cells ( HCECs ) , macrophages , and neutrophils was also assessed . RESULTS : DB04933 tetrasodium significantly inhibited CXC chemokine production in the cornea and development of corneal infiltrates , specifically neutrophils , in response to stimulation with LPS ( O00206 ) , but not Pam(3) DB00151 ( O60603 ) . When the antagonist was applied after LPS stimulation , neutrophil infiltration was also inhibited , although a higher concentration was needed . Furthermore , P10145 production by O00206 - but not O60603 -stimulated HCECs , macrophages and neutrophils was also significantly reduced . Corneal inflammation induced by P. aeruginosa in the presence of tobramycin was found to be dependent on expression of O00206 and MD-2 and is inhibited by eritoran tetrasodium . CONCLUSIONS : DB04933 tetrasodium is a highly effective antagonist of O00206 /MD-2-dependent corneal inflammation . 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 . 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 . 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 . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . PIKKs -- the solenoid nest where partners and kinases meet . The recent structure of a truncated P42345 in a complex with Q9BVC4 has provided a basic framework for understanding all of the phosphoinositide 3-kinase ( PI3K ) -related kinases ( PIKKs ) : P42345 , Q13315 , ATR , Q96Q15 , Q9Y4A5 and DNA-PK . The PIKK kinase domain is encircled by the FAT domain , a helical solenoid that is present in all PIKKs . PIKKs also have an extensive helical solenoid N-terminal to the FAT domain for which there is limited structural information . This N-terminal helical solenoid is essential for binding proteins that associate with the PIKKs to regulate their activity and cellular localization . 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 . 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 . 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 . 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 . Acidic pH induces topoisomerase II-mediated DNA damage . Acidic pH plays an important role in various pathophysiological states and has been demonstrated to be carcinogenic in animal models . Recent studies have also implicated acidic pH in the development of preneoplastic Barrett 's esophagus in human . However , little is known about the molecular mechanism underlying acidic pH-induced carcinogenesis . In the current study , we show that acidic pH , like the topoisomerase II ( P11388 ) poison DB00773 ( demethylepipodophyllotoxin ethylidene-beta-D-glucoside ) , induces tumors in 9,10-dimethyl-1,2-benzanthracene(DMBA)-initiated mice . The following studies in tissue culture models have suggested that acidic pH acts like a P11388 poison to induce P11388 -mediated DNA damage : ( i ) acidic pH induces P11388 -dependent DNA damage signals as evidenced by up-regulation of p53 and DB00133 -139 phosphorylation of P16104 [ a substrate for ataxia telangiectasia mutated ( Q13315 ) Q13315 and Rad3-related ( ATR ) kinases ] ; ( ii ) acidic pH-induced cytotoxicity in tumor cells is reduced in P11388 -deficient cells ; ( iii ) acidic pH increases the mutation frequency of the hypoxanthine phosphoribosyl transferase ( P00492 ) gene in a P11388 -dependent manner ; and ( iv ) acidic pH induces reversible P11388 -mediated DNA strand breaks in vitro . We discuss the possibility that P11388 -mediated DNA damage may contribute to acidic pH-induced carcinogenesis . Biological production of monoethanolamine by engineered Pseudomonas putida P28222 . Pseudomonas putida P28222 was engineered for the production of monoethanolamine ( MEA ) from glucose via the decarboxylation of the central metabolite L-serine , which is catalyzed by the enzyme L-serine decarboxylase ( P18827 ) . The host was first evaluated for its tolerance towards MEA as well as its endogenous ability to degrade this alkanolamine . Growth inhibition was observed at MEA concentrations above 100 mM , but growth was never completely arrested even at 750 mM of MEA . P. putida P28222 was able to catabolize MEA in the absence of ammonia , but deletion of the eutBC genes that encode ethanolamine ammonia-lyase ( EAL ) enzyme sufficed to eliminate this capacity . For the biological production of MEA , the sdc genes from Arabidopsis thaliana ( full-length and a truncated version ) and Volvox carteri were expressed in P. putida P28222 . From 20 mM of glucose , negligible amounts of MEA were produced by P. putida P28222 ΔeutBC expressing the sdc genes from A. thaliana and V. carteri . However , 0.07 mmol of MEA was obtained per g of cell dry weight of P. putida P28222 ΔeutBC expressing the truncated variant of the A. thaliana P18827 . When the medium was supplemented with L-serine ( 30 mM ) , MEA production increased to 1.25 mmol MEA g⁻¹ CDW , demonstrating that L-serine availability was limiting MEA production . Porous polyimide membranes prepared by wet phase inversion for use in low dielectric applications . A wet phase inversion process of polyamic acid ( PAA ) allowed fabrication of a porous membrane of polyimide ( PI ) with the combination of a low dielectric constant ( 1.7 ) and reasonable mechanical properties ( Tensile strain : 8.04 % , toughness : 3.4 MJ/m3 , tensile stress : 39.17 MPa , and young modulus : 1.13 GPa ) , with further thermal imidization process of PAA . PAA was simply synthesized from purified pyromellitic dianhydride ( PMDA ) and 4,4-oxydianiline ( ODA ) in two different reaction solvents such as γ- DB04699 ( Q9BVC4 ) and N-methyl-2-pyrrolidinone ( NMP ) , which produce Mw/ P07237 of 630,000/1.45 and 280,000/2.0 , respectively . The porous PAA membrane was fabricated by the wet phase inversion process based on a solvent/non-solvent system via tailored composition between Q9BVC4 and NMP . The porosity of PI , indicative of a low electric constant , decreased with increasing concentration of Q9BVC4 , which was caused by sponge-like formation . However , due to interplay between the low electric constant ( structural formation ) and the mechanical properties , Q9BVC4 was employed for further exploration , using toluene and acetone vs. DI-water as a coagulation media . Non-solvents influenced determination of the PAA membrane size and porosity . With this approach , insight into the interplay between dielectric properties and mechanical properties will inform a wide range of potential low-k material applications . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Interactions between genetic variants of folate metabolism genes and lifestyle affect plasma homocysteine concentrations in the Boston Puerto Rican population . OBJECTIVE : To investigate genetic and lifestyle factors and their interactions on plasma homocysteine ( Hcy ) concentrations in the Boston Puerto Rican population . DESIGN : Cross-sectional study . Plasma concentrations of Hcy , folate , vitamin B12 and pyridoxal phosphate were measured , and genetic polymorphisms were determined . Data on lifestyle factors were collected in interviews . SETTING : A population survey of health and nutritional measures . SUBJECTS : A total of 994 Puerto Rican men and women residing in the Boston metropolitan area . RESULTS : Smoking status was positively associated with plasma Hcy . Genetic polymorphisms P42898 677C→T , Q04609 1561C→T , Q04609 rs647370 and Q96NT5 928A→G interacted significantly with smoking for Hcy . P42898 1298A→C ( P = 0·040 ) and Q96NT5 928A→G ( P = 0·002 ) displayed significant interactions with alcohol intake in determining plasma Hcy . Subjects with Q96NT5 928GG genotype had significantly higher plasma Hcy concentrations compared with carriers of the A allele ( AA+AG ; P = 0·030 ) among non-drinking subjects . When consuming alcohol , GG subjects had lower plasma Hcy levels compared with AA+AG subjects . Physical activity interacted significantly with Q99707 2756A→G in determining plasma Hcy ( P for interaction = 0·002 ) . Smoking interacted with physical activity for plasma Hcy ( P for interaction = 0·023 ) . CONCLUSIONS : Smoking and drinking were associated plasma Hcy concentrations . Genetic variants involved in folate metabolism further modify the effects of lifestyle on plasma Hcy .

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MH_train_1594

MH_train_1594

interacts_with DB06643?

multiple_choice

P00533 regulates osteoclast differentiation and survival through cross-talking with Q9Y6Q6 signaling . The epidermal growth factor receptor ( P00533 ) functions in various cellular physiological processes such as proliferation , differentiation , and motility . Although recent studies have reported that P00533 signaling is involved in osteoclast recruitment and formation , the molecular mechanism of P00533 signaling for the regulation of osteoclastogenesis remains unclear . We investigated the role of the P00533 in osteoclast differentiation and survival and show that the expression of the P00533 was highly up-regulated by receptor activator of nuclear factor-kappaB ligand ( O14788 ) during osteoclast differentiation . P00533 -specific tyrosine kinase inhibitors and P00533 knockdown blocked O14788 -dependent osteoclast formation , suggesting that P00533 signaling plays an important role in osteoclastogenesis . P00533 inhibition impaired the O14788 -mediated activation of osteoclastogenic signaling pathways , including c-Jun N-terminal kinase ( JNK ) , NF-kappaB , and Akt/protein kinase B ( P31749 ) . In addition , P00533 inhibition in differentiated osteoclasts caused apoptosis through caspase activation . Inhibition of the phosphoinositide-3 kinase ( PI3K ) -Akt/ P31749 pathway and subsequent activation of Q92934 and caspases-9 and -3 may be responsible for the P00533 inhibition-induced apoptosis . The P00533 physically associated with receptor activator of nuclear factor-kappaB ( Q9Y6Q6 ) and Grb2-associated binder 2 ( Gab2 ) . Moreover , O14788 transactivated P00533 . These data indicate that P00533 regulates O14788 -activated signaling pathways by cross-talking with Q9Y6Q6 , suggesting that the P00533 may play a crucial role as a Q9Y6Q6 downstream signal and/or a novel type of Q9Y6Q6 co-receptor in osteoclast differentiation and survival . Gene regulation by hypoxia and the neurodevelopmental origin of schizophrenia . Neurodevelopmental changes may underlie the brain dysfunction seen in schizophrenia . While advances have been made in our understanding of the genetics of schizophrenia , little is known about how non-genetic factors interact with genes for schizophrenia . The present analysis of genes potentially associated with schizophrenia is based on the observation that hypoxia prevails in the embryonic and fetal brain , and that interactions between neuronal genes , molecular regulators of hypoxia , such as hypoxia-inducible factor 1 ( Q9BYW2 ) , and intrinsic hypoxia occur in the developing brain and may create the conditions for complex changes in neurodevelopment . Consequently , we searched the literature for currently hypothesized candidate genes for susceptibility to schizophrenia that may be subject to ischemia-hypoxia regulation and/or associated with vascular expression . Genes were considered when at least two independent reports of a significant association with schizophrenia had appeared in the literature . The analysis showed that more than 50 % of these genes , particularly P31749 , P23560 , O75052 , P32238 , P36544 , P21554 , P21964 , DNTBP1 , Q99259 , Q14832 , P22301 , MLC1 , Q99466 , Q02297 , P43354 / P43354 , O43272 , P78509 , P49798 , Q9NQC3 / Q9NQC3 and P01375 , are subject to regulation by hypoxia and/or are expressed in the vasculature . Future studies of genes proposed as candidates for susceptibility to schizophrenia should include their possible regulation by physiological or pathological hypoxia during development as well as their potential role in cerebral vascular function . 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 . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . Different expression of PGE synthase , P43088 , P01375 , Fas and oxytocin in the bovine corpus luteum of the estrous cycle and pregnancy . Functional differences between the corpus luteum ( CL ) of pregnancy and CL of the cycle in cows were examined . Messenger RNA and protein levels of prostaglandin ( PG ) E synthase ( O14684 ) , PGF2α receptor ( PGFR ) , tumor necrosis factor-α ( P01375 ) and Fas were found to be higher in the CL of pregnancy than in CL of the cycle . DB00107 ( OT ) mRNA and protein levels were lower in the CL of pregnancy . Messenger RNA levels of progesterone receptor ( PR ) , luteinizing hormone receptor ( LHR ) , DB00917 receptor ( PGER ) , P49763 synthase ( P42330 ) , P01375 receptor type I ( TNFRI ) and P01375 receptor type II ( TNFRII ) did not differ between the cycle and pregnancy . DB00917 and PGF2α production by cultured bovine endometrial tissues was decreased by a supernatant derived from the homogenized CL of pregnancy but not by that of the CL of the cycle , suggesting that specific substances in the CL of pregnancy affect endometrial PG production in cows . Collectively , O14684 , PGFR , P01375 , Fas or OT may contribute to differences between the CL of pregnancy and CL of the estrous cycle in cows . cDNA microarray reveals signaling pathways involved in hormones expression of human pituitary . Pituitary , a master gland of neuroendocrine system , secretes hormones that orchestrate many physiological processes , under the regulation of multiple signaling pathways . To investigate the genes involved in hormones expression of human pituitary , homemade cDNA microarray containing 14,800 human genes/ESTs were used to profile the gene expression in both fetal and adult pituitaries . Seven hundred and twelve known genes changed over 2-fold between the both tissues . Of which , 23 genes were changed with hormones expression in aging were confirmed by RT-PCR , not only the known regulators such as Pit1 , P43694 , P11474 , GABA-A , and EMK , but also LOC55884 , P51452 , Q9H307 , and O43598 , which had not been reported to be involved in the hormones expression . Correspondingly , the mRNAs of GH , PRL , P01189 , P01222 , DB00094 -beta , and LH-beta , was increased as much as 6- to 20-fold in adult pituitary than those in fetal pituitary , by real-time quantitative RT-PCR assay . In addition , the mRNAs of signaling pathways , such as DB02527 -PKA-CREB , PI3K-Akt , and PKA- P29323 were further investigated . Of them , it was only DB02527 -PKA-CREB pathway , but not PI3K-Akt and PKA- P29323 have the same expressing pattern as hormones . It suggested that cDNA microarray is highly advantages to profile the differential expressed genes that were involved in hormones expression of human pituitary , but it might ignore some responding proteins regulated posttranscriptionally . Emerging oral drugs for erectile dysfunction . Erectile dysfunction ( ED ) is a common medical condition that affects the sexual life of millions of men worldwide . Many drugs are now available for the treatment of ED , with oral pharmacotherapy representing the first-line option for most patients . DB00203 citrate , an inhibitor of the enzyme phosphodiesterase type 5 ( O76074 ) , is the most widely prescribed oral agent and has a very satisfactory efficacy-safety profile in all patient categories . DB00820 ( DB00820 ; Eli Lilly & Co. , Q9Y6W8 ) and vardenafil ( DB00862 ; Bayer Pharmaceuticals , GlaxoSmithKline ) are new O76074 inhibitors that have recently been approved worldwide . Both have been associated with significant positive efficacy-safety profiles . DB00714 sublingual is a dopamine D1 and D2 receptor agonist , which has been approved for marketing in Europe . It is best selected for treating patients with mild-to-moderate ED , but it is seldom used in clinical practice due to its limited efficacy and side effects , particularly nausea . Patients who do not respond to oral pharmacotherapy or who are unable to use it are appropriate candidates for intracavernosal and intraurethral therapy . The efficacy of second-line treatment is high , but the attrition rate remains significant . For the purpose of this review , clinical and pharmacological analysis focuses on the recent advances in the field of oral therapy , including O76074 inhibitors and sublingual apomorphine . DB09559 in the treatment of advanced non-small cell lung cancer : translation from preclinical to clinical development . INTRODUCTION : Treatment outcomes in unselected patients with advanced NSCLC remain disappointing with platinum-based chemotherapy . The addition of monoclonal antibodies targeting P00533 to standard first-line therapy is a validated strategy and has been associated with statistically significant survival advantage in advanced NSCLC . Necitumunab is a fully human IgG1 monoclonal antibody targeting P00533 , having the potential benefit of lower hypersensitivity reaction risk as compared with cetuximab and also equivalent antibody-dependent cell-mediated cytotoxicity . AREAS COVERED : This paper reviews literature on preclinical and early clinical development of necitumumab that is available in PubMed and published abstracts from conferences , as well as ongoing trials as specified by clinicaltrials.gov . Recently , the Phase III clinical trial evaluating the addition of necitumumab to pemetrexed and cisplatin in non-squamous NSCLC was prematurely closed due to concerns about the increased risk of thromboembolic events in the experimental arm . Accrual in the Phase III trial of necitumumab in combination with gemcitabine and cisplatin in squamous NSCLC is ongoing . EXPERT OPINION : Results of the ongoing large randomized trials will be instrumental in determining the drug 's clinical significance and , with the analysis of potential molecular predictive factors , are expected to bring valuable additions to future therapeutic strategies in NSCLC . Green tea polyphenol epigallocatechin-3-gallate inhibits P01375 -α-induced production of monocyte chemoattractant protein-1 in human umbilical vein endothelial cells . AIMS : DB03823 -3-gallate ( EGCG ) , a major catechin found in green tea , displays a variety of pharmacological properties and recently received attention as a prospective dietary intervention in cardiovascular diseases ( CVD ) . This study was conducted to test the hypothesis that EGCG was able to inhibit tumor necrosis factor-α ( P01375 -α ) -induced production of monocyte chemoattractant protein-1 ( P13500 ) in human umbilical vein endothelial cells ( HUVECs ) and investigated the underlying molecular mechanisms . METHODS : The inhibitory effect of EGCG on P01375 -α-induced expression of P13500 was measured using ELISA and RT-qPCR . The effect of EGCG on P01375 -α-induced nuclear factor-kappa B ( NF-κB ) activation was investigated by western blot and luciferase assays . Monocyte adhesion assay was detected by microscope . RESULTS : EGCG significantly suppressed the P01375 -α-induced protein and mRNA expression of P13500 . Investigation of the mechanism suggested that EGCG suppressed the P01375 -α-mediated NF-κB activation . In addition , the 67-kD laminin receptor ( P08865 ) was involved in EGCG-mediated suppression of P13500 generation . Furthermore , EGCG potently inhibited monocyte adhesion to activated HUVECs . CONCLUSION : EGCG suppresses P01375 -α-induced P13500 expression in HUVECs . This effect was mediated by P08865 and was via the inhibition of NF-κB activation . Our results demonstrated that EGCG might be a possible medicine for CVD prevention and treatment . Glutamate modulators as potential therapeutic drugs in schizophrenia and affective disorders . Severe psychiatric disorders such as schizophrenia are related to cognitive and negative symptoms , which often are resistant to current treatment approaches . The glutamatergic system has been implicated in the pathophysiology of schizophrenia and affective disorders . A key component is the dysfunction of the glutamatergic N-methyl-D-aspartate ( DB01221 ) receptor . Substances regulating activation/inhibition of the DB01221 receptor have been investigated in schizophrenia and major depression and are promising in therapeutic approaches of negative symptoms , cognition , and mood . In schizophrenia , add-on treatments with glycine , D-serine , D-alanine , D-cycloserine , D-amino acid oxidase inhibitors , glycine transporter-1 ( P48067 ) inhibitors ( e.g. , sarcosine , bitopertin ) and agonists ( e.g. , DB05096 ) or positive allosteric modulator ( e.g. , ADX71149 ) of group II metabotropic glutamate receptors ( mGluRs ) have been studied . In major depression , the DB01221 receptor antagonists ( e.g. , ketamine , AZD6765 ) , Q13224 subtype antagonists ( e.g. , traxoprodil , MK-0657 ) , and partial agonists ( e.g. , D-cycloserine , GLYX-13 ) at the glycine site of the DB01221 receptor have been proven to be effective in animal studies and first clinical trials . In addition , clinical studies of Q14416 /3 antagonist BCI-838 ( a prodrug of BCI-632 ( MGS0039 ) ) , Q14416 /3-negative allosteric modulators ( NMAs ) ( e.g. , RO499819 , RO4432717 ) , and P41594 NAMs ( e.g. , AZD2066 , RO4917523 ) are in progress . Future investigations should include effects on brain structure and activation to elucidate neural mechanisms underlying efficacy of these drugs . 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 . Activation of spinal alpha-7 nicotinic acetylcholine receptor attenuates remifentanil-induced postoperative hyperalgesia . The activation of alpha-7 nicotinic acetylcholine receptors ( α7-nAchRs ) are currently being considered as novel therapeutic approaches for managing hyperalgesia in inflammation and chronic neuropathic pain , but the role of a7-nAChRs on opioids induced hyperalgesia remain unknown . The present study investigated the effects of α7-nAChRs selective agonists PHA-543613 and type II positive allosteric modulators ( PAMs ) PNU-120596 in remifentanil induced postoperative hyperalgesia . As the results shown , intrathecal treatment with both α7-nAChRs agonists and type II PAMs could attenuate remifentanil induced hyperalgesia by increasing paw withdrawal mechanical threshold ( PWMT ) and paw withdrawal thermal latency ( PWTL ) . Furthermore , we also investigated the protein level of proinflammatory cytokines and phosphorylation N-methyl-d-aspartate receptor 2B subunit ( p- Q13224 ) in the spinal cord . Our data indicated that activation of α7-nAchRs decreased the proinflammatory cytokines ( P01375 -α , P05231 ) and p- Q13224 protein level in the spinal cord . The depression of the increased levels of proinflammatory cytokines and p- Q13224 after remifentanil treatment may contribute to the anti-hyperalgesia effects of PHA-543613and PNU-120596 via α7-nAChRs . Therefore , our findings demonstrated that α7-nAChRs may be potential candidates for treating opioids induced hyperalgesia . Phenotypic differences between Th1 and Th17 cells and negative regulation of Th1 cell differentiation by Q16552 . Recent evidence from several groups indicates that Q16552 -producing Th17 cells , rather than , as once was thought , P01579 -producing Th1 cells , can represent the key effector cells in the induction/development of several autoimmune and allergic disorders . Although Th17 cells exhibit certain phenotypic and developmental differences from Th1 cells , the extent of the differences between these two T cell subsets is still not fully understood . We found that the expression profile of cell surface molecules on Th17 cells has more similarities to that of Th1 cells than Th2 cells . However , although certain Th1-lineage markers [ i.e. , Q14116 receptor alpha , P49682 , and T cell Ig domain , mucin-like domain-3 ( Q8TDQ0 ) ] , but not Th2-lineage markers ( i.e. , T1/ST2 , Q96D42 , and TIM-2 ) , were expressed on Th17 cells , the intensity of expression was different between Th17 and Th1 cells . Moreover , the expression of P10144 , Q9Y6W8 , programmed death ligand 1 , CD153 , Fas , and O14788 was greater on Th17 cells than on Th1 cells . We found that IL-23 or Q16552 can suppress Th1 cell differentiation in the presence of exogenous IL-12 in vitro . We also confirmed that IL-12 or P01579 can negatively regulate Th17 cell differentiation . However , these cytokines could not modulate such effects on T cell differentiation in the absence of P25054 . DB00996 prevents oxaliplatin-induced central sensitization in the dorsal horn neurons in rats . OBJECTIVES : The present study aims to study the alteration of glutamatergic transmission in the dorsal horn neurons and the effect of gabapentin on oxaliplatin-induced neuropathic pain in rats . MATERIALS AND METHODS : DB00526 ( 5 mg/kg ) or saline was administered to adult male Sprague-Dawley rats . DB00996 ( 60 mg/kg , IP ) or vehicle was injected daily . Mechanical allodynia was assessed using a series of von Frey filaments . The expression of glutamate receptor subunits ( Q13224 and GluR1 ) and brain-derived neurotrophic factor ( P23560 ) was measured in the dorsal horn . The glutamatergic strength was recorded in the spinal cord slices . RESULTS : Administration of oxaliplatin induced significant hyperreactivity to mechanical stimuli in rats , which was attenuated by gabapentin . Significant increase in the expression of P23560 was found in the dorsal horn in rats receiving oxaliplatin , which was prevented by gabapentin . Further studies also observed a significant increase in the expression of GluR1 and Q13224 , as well as enhanced glutamatergic transmission in the dorsal horn neurons in rats treated with oxaliplatin . The upregulation of glutamatergic transmission was significantly reversed by gabapentin . CONCLUSION : These results illustrated an increased expression of P23560 and enhanced glutamatergic transmission in rats with oxaliplatin-induced neuropathic pain , which was markedly attenuated by gabapentin . DB00169 upregulated protein 1 suppresses P01375 -α-induced NF-κB activation in hepatocarcinogenesis . Vitamin D(3) upregulated protein 1 ( Q9H3M7 ) is a candidate tumor suppressor , the expression of which is dramatically reduced in various tumor tissues . In this study , we found that Q9H3M7 expression is suppressed during human hepatic carcinogenesis , and mice lacking Q9H3M7 are much more susceptible to diethylnitrosamine-induced hepatocarcinogenesis compared with wild type mice . Q9H3M7 -deficient tumors proliferated significantly more than wild type tumors and had corresponding changes in the expression of key cell cycle regulatory proteins . In addition , the hepatomitogen-induced response was associated with a considerable increase in the release of P01375 -α and subsequent enhancement of NF-κB activation in Q9H3M7 -deficient mice . When cells were treated with P01375 -α , the Q9H3M7 level was markedly reduced , concomitant with elevated NF-κB activation . Furthermore , the overexpression of Q9H3M7 resulted in the robust suppression of P01375 -α-activated NF-κB activity via association with Q13547 and O15379 . These results indicate that Q9H3M7 negatively regulates hepatocarcinogenesis by suppressing P01375 -α-induced NF-κB activation . The roles of prostanoids in infection and sickness behaviors . A systemic infection in patients causes so-called sickness behaviors , such as fever generation , adrenocorticotropic hormone release , reduced locomotion , loss of social contact , anorexia , and increased sleep . As aspirin-like non-steroidal anti-inflammatory drugs alleviate most of these symptoms , the involvement of prostanoids in the generation of sickness behaviors has been strongly suggested . Prostanoids , consisting of prostaglandins ( PGs ) and thromboxanes ( TXs ) , are a group of lipid mediators formed in response to various stimuli . They include PGD2 , DB00917 , PGF2alpha , DB01240 , and TXA2 . Immediately after synthesis , they are released outside the cells , and exert their actions by binding to a G-protein-coupled rhodopsin-type receptor on the surface of target cells . There are eight types of prostanoid receptors : the Q13258 , four subtypes of PGE receptor , the P43088 , the P43119 , and the TXA receptor . Recently , mice deficient in each of these prostanoid receptors were generated and the examination of these mice in various experimental disease models revealed the important roles of prostaglandin receptor signaling in various pathological processes . In this review , we describe several recent findings that have addressed the mechanisms underlying sickness behaviors and that have identified the critical roles of the signaling of each prostanoid receptor in the elicitation of the stress responses associated with these sickness behaviors . 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 . 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 . Connexin 43 and P29323 regulate tension-induced signal transduction in human periodontal ligament fibroblasts . Periodontal ligament ( PDL ) fibroblasts play an important role in preserving periodontal homeostasis and transmitting mechanical signals to alveolar bone . Connexin 43 ( P17302 ) , a gap junction protein , is essential for bone homeostasis and regulates bone remodeling . However , the function of P17302 in human PDL fibroblast-regulated bone remodeling has not yet been elucidated . In this study , human PDL fibroblasts were exposed to cyclic mechanical tension with a maximum 5 % elongation for different durations . We then examined the expression of signaling molecules related to osteogenesis and osteoclastogenesis at both the mRNA and protein levels as well as the activity of extracellular signal-regulated kinase ( P29323 ) in human PDL fibroblasts after loading . We found that mechanical tension increased P17302 , which further upregulated osteogenic ( e.g. , Q13950 , Osterix , and O00300 ) and down-regulated osteoclastogenic ( e.g. , O14788 ) signaling molecules . Suppressing P17302 gene ( Gja1 ) by siRNA inhibited the increase in osteogenesis-related molecules but enhanced O14788 expression . Similar to P17302 , activated P27361 /2 was also enhanced by mechanical tension and suppressed by P17302 siRNA . Inhibition of P27361 /2 signaling using PD98059 reduced the tension-regulated increase in osteogenesis-related molecules but enhanced that of osteoclastogenesis-related ones . These findings suggest that cyclic tension may involve into the osteogenic or osteoclastogenetic differentiation potential of human PDL fibroblasts via the P17302 - P27361 /2 signaling pathway . 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 .

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P0DN86 activates P29323 and AKT kinases in cancer cells via P22888 -independent mechanism . Expression of human chorionic gonadotropin free beta subunit ( hCGβ ) and its hyperglycosylated variant ( hCGβ-H ) is a phenomenon confirmed for tumors of different origin . Despite numerous studies , the mechanism of hCGβ action in cancer remains unknown especially that not all tumors secreting hCGβ express the receptor for human chorionic gonadotropin ( P22888 ) . In the presented study , we verified the hypothesis of hCGβ potential to activate signaling pathways involving extracellular signal-regulated kinase ( P29323 ) and protein kinase B ( AKT ) kinases with and without the contribution of P22888 . To achieve this goal , human ovarian carcinoma cells OVCAR-3 expressing P22888 and SKOV-3 not expressing P22888 were either transfected with a vector coding for hCGβ or stimulated with recombinant hCGβ and the level of pERK and pAKT was measured . The results of the experiments showed that hCGβ action leads to the increase in P29323 and AKT kinases phosphorylation in cancer cells and indicate that these biological effects can be achieved independently of P22888 presence . The study also demonstrated that the presence of the receptor is a key factor influencing the magnitude of cells ' response . [ Fibroblast growth factor receptor and achondroplasia ] . P22607 ( P22607 ) has been establishing its position in growth plate cartilage after the identification as a responsible gene for achondroplasia . The major pathway of the pathogenesis in achondroplasia is the suppression of parathyroid hormone-related protein ( P12272 ) -parathyroid hormone receptor ( Q03431 ) system , which is mainly mediated by extracellular signal regulated kinase ( P29323 ) activation induced by constitutive active P22607 . However , intracellular signaling system in P22607 is complex and the molecular pathogenesis of achondroplasia and related disorders has not been fully clarified . In this review , I summarized recent consensus in the pathogenesis of P22607 related chondrodysplasia . 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 . Epigenetic regulation of protein tyrosine phosphatase Q05209 in triple-negative breast cancer . AIMS : The present study showed that the expression of Q05209 is epigenetically regulated . DB00928 ( 5-Azac ) , a DNA hypomethylating agent , significantly increased the expression of Q05209 at low concentrations ( 1μM and 2.5μM ) and decreased the expression of Q05209 at 5μM in the MDA-MB-231 and BT-549 triple-negative breast cancer cell lines . MAIN METHODS : Human MCF-7 , MDA-MB-231 and BT-549 cells were exposed to different concentrations of 5-Azac for 24 and 48h . RT-PCR was performed to determine the mRNA expression of Q05209 , P12830 and miRNA-124 . Western blotting was performed to assess the protein expression of various proteins , including Q05209 , P12830 , P26358 and PARP . KEY FINDINGS : 5-Azac , a DNA hypomethylating agent , significantly increased the expression of Q05209 at low concentrations ( 1μM and 2.5μM ) and decreased Q05209 expression at 5μM . We provide the first evidence that Q05209 expression is epigenetically regulated and that it is up-regulated at a lower dose of a P26358 inhibitor in MDA-MB-231 and BT-549 cells . Interestingly , the levels of miRNA-124 were increased only at 5μM , the concentration at which Q05209 expression was suppressed . SIGNIFICANCE : To the best of our knowledge , this is the first report that highlights the therapeutic potential of low-dose 5-Azac for the treatment of TNBC . Therefore , 5-Azac , an agent that has already been tested in acute myeloid leukemia , may be more effective at lower doses for the treatment of triple-negative breast cancer . P06401 activation of extranuclear signaling pathways in regulating p53 expression in vascular endothelial cells . We previously showed that progesterone ( P4 ) inhibited the proliferation of human umbilical vein endothelial cells ( HUVECs ) through a p53-dependent pathway . Now we investigated further the molecular mechanism underlying the hormone activity . In cultured HUVECs , P4 increased the protein levels of phosphorylated Src ( p-Src ) , P04049 , and P29323 . The levels of p-Src and p-Src-progesterone receptor complex in HUVECs were increased by P4 treatment . These effects were blocked by pretreatment with a progesterone receptor antagonist , DB00834 . The P4-induced increase in p53 transactivity was abolished by pretreatment with Src kinase inhibitors . Moreover , administration with cSrc antisense oligonucleotide prevented the P4-induced increases of the levels of p53 mRNA and protein . These data suggest that P4-induced up-regulation of p53 might be mediated through activation of cSrc . Pretreatment with Src kinase inhibitors also prevented P4-induced membrane translocation of Kras and increases of the protein levels of phosphorylated Raf and phosphorylated P29323 . Transfection with dominant-negative P28482 prevented the P4-induced increases of protein level and promoter activity of p53 and a decrease of thymidine incorporation . P4 also increased nuclear factor-κB ( NF-κB ) nuclear translocation and NF-κB binding onto the p53 promoter . These effects were abolished by pretreatment with P29323 inhibitors . The P4-induced up-regulation of the p53 promoter activity was prevented by preadministration with dominant-negative P28482 or NF-κB inhibitors . Taken together , our data suggest that the cSrc/Kras/ P04049 / P28482 /NF-κB signaling pathway contributes to the P4-induced up-regulation of p53 in HUVECs . These findings highlight progesterone receptor activation of extranuclear signaling pathways in regulating p53 and cell cycle progression in HUVECs . P06401 -B regulation of insulin-like growth factor-stimulated cell migration in breast cancer cells via insulin receptor substrate-2 . Progesterone action contributes to the signaling of many growth factor pathways relevant to breast cancer tumor biology , including the insulin-like growth factor ( IGF ) system . Previous work has shown that insulin receptor substrate-2 ( Q9Y4H2 ) but not P35568 levels were regulated by progestin in progesterone receptor-B ( PR-B ) isoform expressing MCF-7 cells ( C4-12 PR-B ) . Furthermore , type 1 IGF receptor ( P08069 ) signaling via Q9Y4H2 correlated with the increased cell migration observed in a number of breast cancer cell lines . Consequently , in this study , we examined whether the elevation of Q9Y4H2 protein induced by progestin was sufficient to promote P05019 -stimulated cell motility . Treatment of C4-12 PR-B cells with progestin shifted the balance of phosphorylation from P35568 to Q9Y4H2 in response to P05019 . This shift in Q9Y4H2 activation was associated with enhanced migration in C4-12 PR-B cells pretreated with progestin , but had no effect on cell proliferation or survival . Treatment of C4-12 PR-B cells with DB00834 , an antiprogestin , inhibited IGF-induced cell migration . Attenuation of Q9Y4H2 expression using small interfering RNA resulted in decreased IGF-stimulated motility . In addition , Q9Y4H2 knockdown resulted in an abrogation of P31749 /Akt phosphorylation but not mitogen-activated protein kinase . Consequently , LY294002 , a phosphoinositide-3-kinase inhibitor , abolished IGF-induced cell motility in progestin-treated C4-12 PR-B cells . These data show a role for the PR in functionally promoting growth factor signaling , showing that levels of P41252 proteins can determine IGF-mediated biology , PR-B signaling regulates Q9Y4H2 expression , and that Q9Y4H2 can mediate IGF-induced cell migration via phosphoinositide-3-kinase in breast cancer cells . 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 . CP-101,606 , a potent neuroprotectant selective for forebrain neurons . The neuroprotective activity of ( 1S,2S ) -1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol ( CP-101,606 ) , an N-methyl-D-aspartate ( DB01221 ) receptor antagonist structurally similar to ( (+/-)-(R*,S*)-alpha-(4-hydroxyphenyl)-beta-methyl-4-(phenylmethyl)-1-++ +piperidineethanol ( ifenprodil ) , was investigated in neurons in primary culture . CP-101,606 potently and efficaciously protected hippocampal neurons from glutamate toxicity but was > 900-fold less effective for cerebellar granule neurons . The neuroprotective activity in the hippocampal neurons is mediated through a high affinity binding site distinct from the agonist and thienylcyclohexylpiperidine ( DB01520 ) binding sites of the DB01221 receptor . Autoradiography indicates the CP-101,606 binding site is localized in forebrain , most notably in hippocampus and the outer layers of cortex . The functional selectivity for hippocampal neurons , forebrain localization of binding sites , and structural relation to ifenprodil suggest that CP-101,606 is an DB01221 antagonist highly selective for Q13224 subunit containing receptors . Endocannabinoid-dependent neocortical layer-5 LTD in the absence of postsynaptic spiking . Long-term depression ( LTD ) was induced in neocortical layer 5 pyramidal connections by pairing presynaptic firing with subthreshold postsynaptic depolarization ( dLTD ) or via a spike-timing protocol ( tLTD ) . Like tLTD , dLTD reduced short-term depression and increased the coefficient of variation consistent with a presynaptic site of expression . Also like tLTD , dLTD was blocked by P21554 cannibinoid receptor blockade and required activation of presumably presynaptic Q13224 -containing N-methyl-D-aspartate receptors . The two forms of LTD had identical magnitudes and time courses and occluded one another , and neither depended on frequency . Finally , dLTD shares with tLTD the asymmetric temporal window of induction . In conclusion , the types of LTD induced by these two protocols are indistinguishable , suggesting that the mechanism that underlies tLTD paradoxically does not require postsynaptic spiking : The subthreshold postsynaptic depolarizations of dLTD appear to fully substitute for postsynaptic spiking . Molecular components and functions of the endocannabinoid system in mouse prefrontal cortex . BACKGROUND : Cannabinoids have deleterious effects on prefrontal cortex ( P27918 ) -mediated functions and multiple evidences link the endogenous cannabinoid ( endocannabinoid ) system , cannabis use and schizophrenia , a disease in which P27918 functions are altered . Nonetheless , the molecular composition and the physiological functions of the endocannabinoid system in the P27918 are unknown . METHODOLOGY/PRINCIPAL FINDINGS : Here , using electron microscopy we found that key proteins involved in endocannabinoid signaling are expressed in layers v/vi of the mouse prelimbic area of the P27918 : presynaptic cannabinoid P21554 receptors ( CB1R ) faced postsynaptic P41594 while diacylglycerol lipase alpha ( Q9Y4D2 ) , the enzyme generating the endocannabinoid 2-arachidonoyl-glycerol ( 2-AG ) was expressed in the same dendritic processes as P41594 . Activation of presynaptic CB1R strongly inhibited evoked excitatory post-synaptic currents . Prolonged synaptic stimulation at 10Hz induced a profound long-term depression ( LTD ) of layers V/VI excitatory inputs . The endocannabinoid -LTD was presynaptically expressed and depended on the activation of postsynaptic P41594 , phospholipase C and a rise in postsynaptic Ca(2+) as predicted from the localization of the different components of the endocannabinoid system . Blocking the degradation of 2-AG ( with Q76M96 602 ) but not of anandamide ( with Q76M96 597 ) converted subthreshold tetanus to LTD-inducing ones . Moreover , inhibiting the synthesis of 2-AG with DB01083 , blocked endocannabinoid-mediated LTD . All together , our data show that 2-AG mediates LTD at these synapses . CONCLUSIONS/SIGNIFICANCE : Our data show that the endocannabinoid -retrograde signaling plays a prominent role in long-term synaptic plasticity at the excitatory synapses of the P27918 . Alterations of endocannabinoid -mediated synaptic plasticity may participate to the etiology of P27918 -related pathologies . Suppression of P21554 cannabinoid receptor by lentivirus mediated small interfering RNA ameliorates hepatic fibrosis in rats . It is recognized that endogenous cannabinoids , which signal through P21554 receptors in hepatic stellate cells ( HSCs ) , exert a profibrotic effect on chronic liver diseases . In this study , we suppressed P21554 expression by lentivirus mediated small interfering RNA ( P21554 -RNAi-LV ) and investigated its effect on hepatic fibrosis in vitro and in vivo . Our results demonstrated that P21554 -RNAi-LV significantly inhibited P21554 expression , and suppressed proliferation and extracellular matrix production in HSCs . Furthermore , P21554 -RNAi-LV ameliorated dimethylnitrosamine induced hepatic fibrosis markedly , which was associated with the decreased expression of mesenchymal cell markers smooth muscle α-actin , vimentin and snail , and the increased expression of epithelial cell marker P12830 . The mechanism lies on the blockage of Smad signaling transduction induced by transforming growth factor β1 and its receptor TGF-β RII . Our study firstly provides the evidence that P21554 -RNAi-LV might ameliorate hepatic fibrosis through the reversal of epithelial-to-mesenchymal transition ( EMT ) , while the P21554 antagonists AM251 had no effect on epithelial-mesenchymal transitions of HSCs . This suggests that P21554 is implicated in hepatic fibrosis and selective suppression of P21554 by small interfering RNA may present a powerful tool for hepatic fibrosis treatment . Phosphodiesterase 4 inhibitor cilomilast inhibits fibroblast-mediated collagen gel degradation induced by tumor necrosis factor-alpha and neutrophil elastase . Tissue destruction , resulting in emphysema , can be a consequence of several pathologic processes . The current study evaluated the effects of the phosphodiesterase (PDE)4 inhibitor , cilomilast , and other PDE inhibitors on the ability of fibroblasts to degrade extracellular matrix . Using the three-dimensional collagen gel culture system , fibroblasts ( HFL-1 ) were cultured with tumor necrosis factor ( P01375 ) -alpha , known to induce matrix metalloproteinase ( MMP ) release , and/or neutrophil elastase ( NE ) , which can induce MMP activation . On Day 4 , gels containing P01375 and NE were significantly degraded ( 20.8 +/- 2.9 % of original collagen content ) . DB03849 ( 10 micro M ) inhibited this degradation ( 84.4 +/- 8.4 % ) . DB01427 , a PDE3 inhibitor , and zaprinast , a O76074 inhibitor , had no effect . Gelatin zymography and immunoblotting revealed that fibroblasts cultured with P01375 released increased amounts of latent P03956 and -9 . The addition of NE resulted in the conversion of P03956 and -9 to their active forms , indicative of collagen degradation . DB03849 inhibited the release of P03956 and -9 , as well as conversion of P03956 to its active form . Using real-time PCR analysis , cilomilast 's effect on P03956 release was not associated with the proteinase 's mRNA expression , suggesting that the inhibition of release is regulated at the post-transcriptional level . These results suggest that cilomilast may be a potentially effective therapeutic agent in diseases characterized by excessive tissue destruction , such as emphysema . Glutamatergic fine tuning with P10109 -10059 : a novel therapeutic approach for migraine ? IMPORTANCE OF THE FIELD : Migraine is an episodic , substantially inherited brain disorder affecting 15 % of adults in Western Europe and North America , and is one of the commonest reasons for patients to see their physicians . While the World Health Organization considers that severe migraine can be as disabling as quadriplegia , unfortunately the condition remains undertreated . Until the 1990s , specific migraine therapies were limited to ergot derivatives . AREAS COVERED IN THIS REVIEW : The triptans , serotonin 5-HT(1B/1D) receptor agonists , revolutionized the acute management of migraine patients . However , although the triptans are generally effective and safe , not all patients can take them and many do not respond especially to oral therapies . Recently , progress has been made on the therapeutic front , particularly with new acute treatments . This review will focus on the therapeutic potential of DB05070 , a metabotropic glutamate receptor 5 , negative allosteric modulator ( P41594 NAM ) , in migraine . Data from a proof-of-concept study in episodic migraineurs demonstrated a significant improvement following acute treatment . A large European multicenter , randomized , double-blind , placebo-controlled , parallel-group , dose-ranging study is currently investigating the efficacy , safety and tolerability of the compound for migraine prevention . WHAT THE READER WILL GAIN : The reader will have the basic principles of migraine management and the potential for glutamate-targeted approaches . TAKE HOME MESSAGE : Targeting glutamatergic transmission in migraine may provide a novel preventive therapy that is effective and well-tolerated . 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 . P04150 -mediated suppression of activator protein-1 activation and matrix metalloproteinase expression after spinal cord injury . Post-traumatic inflammatory reaction may contribute to progressive tissue damage after spinal cord injury ( SCI ) . Two key transcription factors , nuclear factor kappaB ( NF-kappaB ) and activator protein-1 ( AP-1 ) , are activated in inflammation . An increase in NF-kappaB binding activity has been shown in the injured spinal cord . We report activation of AP-1 after SCI . Electrophoretic mobility shift assay showed that AP-1 binding activity increased after SCI , starting at 1 hr , peaking at 8 hr , and declining to basal levels by 7 d . Methylprednisolone ( MP ) is the only therapeutic agent approved by the Food and Drug Administration for treating patients with acute traumatic SCI . MP reduced post-traumatic AP-1 activation . DB00834 , a glucocorticoid receptor ( GR ) antagonist , reversed MP inhibition of AP-1 activation . Immunostaining showed an increase in the expression of the Fos-B and c-Jun components of AP-1 in the injured cord . A c-fos antisense oligodeoxynucleotide ( ODN ) inhibited AP-1 , but not NF-kappaB , activation after SCI . AP-1 and NF-kappaB can transactivate genes encoding matrix metalloproteinase-1 ( P03956 ) and P14780 . Western blotting and immunostaining show increased expression of P03956 and P14780 in the injured cord . MP inhibited P03956 and P14780 expression after SCI . DB00834 reversed this MP effect . The c-fos antisense ODN , however , failed to suppress P03956 or P14780 expression . These findings demonstrate that MP may suppress post-traumatic inflammatory reaction by inhibiting both the AP-1 and NF-kappaB transcription cascades via a GR mechanism . Expression of inflammatory genes such as P03956 and P14780 that are transactivated jointly by AP-1 and NF-kappaB may not be suppressed by inhibiting only AP-1 activity . Acrodysostosis . Acrodysostosis refers to a group of rare skeletal dysplasias that share in common characteristic clinical and radiological features including brachydactyly , facial dysostosis , and nasal hypoplasia . In the past , the term acrodysostosis has been used to describe patients with heterogeneous phenotypes , including , in some cases , patients that today would be given alternative diagnoses . The recent finding that mutations impairing the DB02527 binding to P10644 are associated with " typical " acrodysostosis and hormonal resistance initiates the era where this group of disorders can be categorized on a genetic basis . In this review , we will first discuss the clinical , radiologic , and metabolic features of acrodysostosis , emphasizing evidence that several forms of the disease are likely to exist . Second , we will describe recent results explaining the pathogenesis of acrodysostosis with hormonal resistance ( ADOHR ) . Finally , we will discuss the similarities and differences observed comparing patients with ADOHR and other diseases resulting from defects in the Q03431 signaling pathway , in particular , pseudohypoparathyroidism type 1a and pseudopseudohypoparathyroidism . Expression of genes related to prostaglandin synthesis or signaling in human subcutaneous and omental adipose tissue : depot differences and modulation by adipogenesis . OBJECTIVES : ( 1 ) To examine depot-specific DB00917 and PGF2α release and mRNA expression of enzymes or receptors involved in PG synthesis or signaling in human adipose tissues ; ( 2 ) to identify changes in expression of these transcripts through preadipocyte differentiation ; and ( 3 ) to examine associations between adipose tissue mRNA expression of these transcripts and adiposity measurements . METHODS : Fat samples were obtained surgically in women . DB00917 and PGF2α release by preadipocytes and adipose tissue explants was measured . Expression levels of mRNA coding for enzymes or receptors involved in PG synthesis or signaling were measured by RT-PCR . RESULTS : Cultured preadipocytes and explants from omental fat released more DB00917 and PGF2α than those from the subcutaneous depot and the corresponding transcripts showed consistent depot differences . Following preadipocyte differentiation , expression of P53816 and P43115 mRNA was significantly increased whereas P23219 , P35354 , Q16647 , and O14684 mRNA abundance were decreased in both compartments ( P ≤ 0.01 for all ) . Transcripts that were stimulated during adipogenesis were those that correlated best with adiposity measurements . CONCLUSION : Cells from the omental fat compartment release more DB00917 and PGF2α than those from the subcutaneous depot . Obesity modulates expression of PG-synthesizing enzymes and PG receptors which likely occurs through adipogenesis-induced changes in expression of these transcripts . Tumour necrosis factor inhibitors : risks and benefits in patients with rheumatoid arthritis . Rheumatoid arthritis ( RA ) is the most common form of inflammatory arthritis and can , if left untreated , result in significant disability and early death . It is also associated with large direct and indirect costs to the individual and to society . Early and aggressive disease modifying anti-rheumatic drug ( DMARD ) treatment of patients at risk of erosive disease has improved the outcome in the majority , but not all , RA patients . Tumour necrosis factor ( P01375 ) appears to be a key mediator of the inflammatory and destructive process in RA , and consequently inhibitors of P01375 action have been tested in randomized controlled trials in patients with RA . The results of these studies have suggested that P01375 inhibitors are potent DMARD particularly when combined with methotrexate . They appear well tolerated with the commonest adverse events related to their parenteral route of administration , and the serious but rare side-effects being various infections , notably tuberculosis , multiple sclerosis , and worsening of cardiac failure . Treatment costs are high and range from $ 15 000 to $ 25 000 per patient per year . DB00005 , adalimumab and infliximab have recently been subsidised under the Pharmaceutical Benefits Scheme in Australia for patients with severe DMARD-resistant RA . The availability of P01375 inhibitors in RA represents a significant advance in the treatment of patients with severe RA . P04150 signaling in a bronchial epithelial cell line . Glucocorticoids are an effective anti-inflammatory therapy for the treatment of asthma . The anti-inflammatory effects of glucocorticoids may be due to the inhibition of transcription factors that regulate cytokine synthesis . Because of the potential role of the bronchial epithelium in asthmatic inflammation and the possibility that this cell may be the main target of inhaled glucocorticoids , we have characterized glucocorticoid receptors ( GR ) and GR signaling in the human bronchial epithelial cell line BEAS-2B . Western blot analysis and radioligand binding studies demonstrated that BEAS-2B cells have functional GR that bind to dexamethasone ( DB00514 ) ( dissociation constant = 5.6 nM and maximal density of binding sites = 228 +/- 3.3 fmol/mg protein ) . GR were activated by DB00514 as assessed using a glucocorticoid-responsive reporter plasmid . Transfection of BEAS-2B cells with an activator protein-1 ( AP-1 ) reporter construct followed by 12-O-tetradecanoylphorbol-13-acetate ( TPA ) treatment resulted in a fivefold induction of reporter gene activity . Transfection with a nuclear factor ( NF ) -kappa B reporter construct followed by tumor necrosis factor-alpha ( P01375 ) treatment resulted in a 10-fold induction of reporter gene activity . DB00514 ( 10(-7) M ) markedly repressed both the induced AP-1 and NF-kappa B activity . The GR antagonist DB00834 inhibited the repressive effect of DB00514 on P01375 -induced NF-kappa B activity by 81 % but only counteracted the repressive effect of DB00514 on TPA-induced AP-1 activity by 43 % . These studies demonstrate that cross-signaling between AP-1 and NF-kappa B with GR may explain the anti-inflammatory properties of glucocorticoids in airway epithelial cells . DB00044 releasing hormone , estrogen , and progesterone receptors in canine mammary lesions and tumor cell lines . Nineteen canine mammary lesions were analyzed for estrogen receptor ( ER ) , progesterone receptor ( PR ) and luteinizing hormone releasing hormone receptor ( P22888 ) content . In addition , nine previously established canine mammary tumor cell lines with known ER and PR were analyzed for P22888 . The incidence of receptors in the mammary tumor lesions was 21 % for P22888 , 10 % for ER and 30 % for PR . Statistical correlation was not observed between receptor status and diagnosis of malignant , benign , or hyperplastic lesions . A relationship between P22888 and ER and PR content of canine mammary lesions or cell lines was not evident . The presence of functional hormone receptors offers opportunity for hormonal treatment of mammary cancer which may not be completely treatable by surgery . The observation of P22888 in canine mammary tumors may offer therapeutic interventions other than surgery for mammary tumors unresponsive to antiestrogens .

MH_train_1596

MH_train_1596

MH_train_1596

interacts_with DB00072?

multiple_choice

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 . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . A RANTES-antibody fusion protein retains antigen specificity and chemokine function . The successful eradication of cancer cells in the setting of minimal residual disease may require targeting of metastatic tumor deposits that evade the immune system . We combined the targeting flexibility and specificity of mAbs with the immune effector function of the chemokine RANTES to target established tumor deposits . We describe the construction of an Ab fusion molecule with variable domains directed against the tumor-associated Ag P04626 /neu , linked to sequences encoding the chemokine RANTES ( RANTES.her2.IgG3 ) . RANTES is a potent chemoattractant of T cells , NK cells , monocytes , and dendritic cells , and expression of RANTES has been shown to enhance immune responses against tumors in murine models . RANTES.her2.IgG3 fusion protein bound specifically to P04626 /neu Ag expressed on EL4 cells and on SKBR3 breast cancer cells as assayed by flow cytometry . RANTES.her2.IgG3 could elicit actin polymerization of THP-1 cells and transendothelial migration of primary T lymphocytes . RANTES.her2.IgG3 prebound to SKBR3 cells also facilitated migration of T cells . RANTES.her2.IgG3 bound specifically to the P51681 chemokine receptor , as demonstrated by flow cytometry , and inhibited HIV-1 infection via the P51681 coreceptor . RANTES.her2.IgG3 , alone or in combination with other chemokine or cytokine fusion Abs , may be a suitable reagent for recruitment and activation of an expanded repertoire of effector cells to tumor deposits . Short and long access to cocaine self-administration activates tyrosine phosphatase P54829 and attenuates GluN expression but differentially regulates GluA expression in the prefrontal cortex . RATIONALE : Dephosphorylation of extracellular signal-regulated kinase ( P29323 ) and cyclic AMP response element binding protein ( CREB ) in the dorsomedial prefrontal cortex ( dmPFC ) at the end of short access ( ShA ) cocaine self-administration is implicated in cocaine seeking . However , what receptors and phosphatases mediate this effect and whether P29323 /CREB and related phospho-proteins in the dmPFC react similarly during early withdrawal from long access ( LgA ) cocaine self-administration are unknown . OBJECTIVES : The effects of ShA vs. LgA cocaine self-administration on the phosphorylation of protein phosphatase 2A ( PP2A ) and striatal-enriched protein tyrosine phosphatase ( P54829 ) , as well as GluN and GluA receptor subtype expression in the dmPFC during early withdrawal , were compared . METHODS : Rats self-administered cocaine or received saline during 2- or 6-h daily sessions for 10-11 days . Two hours after the final session , the dmPFC was dissected out and processed for immunoblotting . RESULTS : Similar to previous findings after ShA cocaine , phospho- P29323 and phospho-CREB in the dmPFC were decreased after LgA cocaine . Cocaine elevated phospho-PP2A ( deactivation ) and decreased phospho- P54829 ( activation ) in both ShA and LgA cocaine rats . Q05586 , Q13224 , and phospho- Q13224 Tyr1472 in the dmPFC were decreased after ShA and LgA cocaine . Further , a significant reduction of P42262 , P42261 , and phospho- P42261 Ser845 was found only in LgA rats . CONCLUSIONS : Activation of phospho- P54829 may underlie P29323 and CREB dephosphorylation in the dmPFC as well as internalization and degradation of GluN complexes during early withdrawal from both ShA and LgA cocaine self-administration , whereas differential alteration of AMPA receptor subunits after ShA and LgA cocaine self-administration depends on cocaine intake . Short-term biomarker modulation prevention study of anastrozole in women at increased risk for second primary breast cancer . The selective estrogen receptor modulators ( SERM ) , Tamoxifen and raloxifen reduce risk breast cancer . Patient acceptance of SERMs for breast cancer prevention is low due to toxicities . New agents with a better toxicity profile are needed . P11511 inhibitors ( AI ) reduce the risk of contralateral breast cancer and risk of new breast cancer in high risk women . However , the mechanism by which AIs reduce breast risk is not known . Surrogate biomarkers are needed to evaluate the effect of preventive agents . The objective of this prospective short-term prevention study was to evaluate the effect of anastrozole on biomarkers in breast tissue and serum of women at increased risk for developing a contralateral breast cancer . Women with a history of stage I , II breast cancer who started anastrozole for standard adjuvant treatment were eligible . Patients underwent baseline fine needle aspiration of the unaffected breast and serum collection for biomarker analysis before starting anastrozole at 1 mg per oral/day and again at 6 months . Biomarkers included changes in cytology , insulin-like growth factor 1 ( DB01277 ) , P08833 ( P08833 ) , and P17936 . Thirty-seven patients were enrolled . There was a significant modulation in serum P08833 levels between pre- and postsamples ( P = 0.02 ) . No change was observed in DB01277 , P17936 , and breast cytology.We showed a significant modulation of P08833 levels with six months anastrozole . DB01217 is currently being studied as a prevention agent in a large phase III trial and our results provide support for continued evaluation of P08833 as a surrogate endpoint biomarker in prospective breast chemoprevention studies . P01308 -like growth factor-I receptor signaling and resistance to trastuzumab ( Herceptin ) . BACKGROUND : DB00072 ( Herceptin ) , an anti- P04626 /neu receptor monoclonal antibody that inhibits growth of ErbB2-overexpressing breast cancer , is used to treat such cancers . Development of resistance to trastuzumab , however , is common . We investigated whether insulin-like growth factor-I ( P05019 ) , which activates cell survival signals , interferes with the growth-inhibitory action of trastuzumab . METHODS : MCF-7/ P04626 -18 and SKBR3 human breast cancer models were used to assess cell proliferation , colony formation in soft agar , and cell cycle parameters . Throughout , we used trastuzumab at a dose of 10 microg/mL and P05019 at a dose of 40 ng/mL . All statistical tests were two-sided . RESULTS : DB00072 inhibited the growth of MCF-7/ P04626 -18 cells , which overexpress P04626 /neu receptors and express P05019 receptors ( IGF-IRs ) , only when IGF-IR signaling was minimized . For example , in 1 % fetal bovine serum ( FBS ) , trastuzumab reduced cell proliferation by 42 % ( P =.002 ) ; however , in 10 % FBS or P05019 , trastuzumab had no effect on proliferation . In SKBR3 cells , which overexpress P04626 /neu receptor but express few IGF-IRs , trastuzumab reduced proliferation by 42 % ( P =.008 ) regardless of P05019 concentration . When SKBR3 cells were genetically altered to overexpress IGF-IRs and cultured with P05019 , trastuzumab had no effect on proliferation . However , the addition of IGF-binding protein-3 , which decreased IGF-IR signaling , restored trastuzumab-induced growth inhibition . CONCLUSIONS : In breast cancer cell models that overexpress P04626 /neu , an increased level of IGF-IR signaling appears to interfere with the action of trastuzumab . Thus , strategies that target IGF-IR signaling may prevent or delay development of resistance to trastuzumab . 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 . Enhanced sensitization to taxol-induced apoptosis by herceptin pretreatment in ErbB2-overexpressing breast cancer cells . The recombinant humanized anti-ErbB2/ P04626 monoclonal antibody Herceptin ( DB00072 ) has been shown to significantly enhance the tumoricidaleffects of antitumor drugs such as paclitaxel ( DB01229 ) in patients with ErbB2-overexpressing breast cancers . Here , we investigated the molecular mechanisms by which Herceptin enhances the antitumor effects of DB01229 . Because activation of p34(Cdc2) is required for DB01229 -induced apoptosis and because overexpression of ErbB2 blocks DB01229 -induced apoptosis by inhibiting p34(Cdc2) activation , we studied the effect of Herceptin treatment on p34(Cdc2) kinase activation and apoptosis in DB01229 -treated human breast carcinoma cell lines MDA-MB-435 , SKBr3 , MDA-MB-453 , and 435.eB , which is an ErbB2 transfectant of MDA-MB-435 . Herceptin treatment down-regulated ErbB2 , reduced the inhibitory phosphorylation of Cdc2 on DB00135 -15 , and down-regulated the expression of P38936 (Cip1) , a Cdc2 inhibitor . Herceptin plus DB01229 treatment led to higher levels of p34(Cdc2) kinase activity and apoptosis in ErbB2-overexpressing breast cancer cells , which is likely attributable to inhibition of Cdc2- DB00135 -15 phosphorylation and P38936 (Cip1) expression . Because significant dephosphorylation of Cdc2- DB00135 -15 and down-regulation of P38936 (Cip1) occur at least 24 h after Herceptin treatment , we investigated whether 24 h Herceptin pretreatment will render ErbB2-overexpressing breast cancer cells more sensitive to DB01229 -induced apoptosis compared with the simultaneous treatment of Herceptin plus DB01229 . Indeed , Herceptin pretreatment increased DB01229 -induced apoptosis and cytotoxicity in vitro and more effectively inhibited the growth of tumor xenografts with enhanced in vivo apoptosis . Thus , Herceptin treatment of ErbB2-overexpressing cells can inhibit ErbB2-mediated Cdc2- DB00135 -15 phosphorylation and P38936 (Cip1) up-regulation , which allows effective p34(Cdc2) activation and induction of apoptosis upon DB01229 treatment . Herceptin pretreatment renders ErbB2-overexpressing breast cancers more susceptible to DB01229 -induced cell death , which may have important clinical therapeutic implications . Lipoplex mediated silencing of membrane regulators ( P15529 , P08174 and P13987 ) enhances complement-dependent anti-tumor activity of trastuzumab and pertuzumab . The therapeutic potential of anticancer antibodies is limited by the resistance of tumor cells to complement-mediated attack , primarily through the over-expression of membrane complement regulatory proteins ( mCRPs : P15529 , P08174 and P13987 ) . DB00072 , an anti- P04626 monoclonal antibody , approved for the treatment of P04626 -positive breast and gastric cancers , exerts only minor complement-mediated cytotoxicity ( CDC ) . DB06366 is a novel anti- P04626 monoclonal antibody , which blocks P04626 dimerization with other ligand-activated HER family members . Here , we explored the complement-mediated anti-tumor effects of trastuzumab and pertuzumab on P04626 -positive tumor cells of various histological origins . Delivery of chemically stabilized anti-mCRP siRNAs using cationic lipoplexes , AtuPLEXes , to P04626 -over-expressing BT474 , SK-BR-3 ( breast ) , SKOV3 ( ovarian ) and Calu-3 ( lung ) cancer cells reduced mCRPs expression by 85-95 % . Knockdown of individual complement regulators variably led to increased CDC only upon combined treatment with trastuzumab and pertuzumab . The combined down-regulation of all the three regulators augmented CDC by 48 % in BT474 , 46 % in SK-BR-3 cells , 78 % in SKOV3 cells and by 30 % in Calu-3 cells and also increased complement-induced apoptosis and caspase activity on mCRP neutralized tumor cells . In addition , antibody-induced P01024 opsonization of tumor cells was significantly enhanced after mCRP silencing and further augmented tumor cell killing by macrophages . Our findings suggest that siRNA-induced inhibition of complement regulator expression clearly enhances complement- and macrophage-mediated anti-tumor activity of trastuzumab and pertuzumab on P04626 -positive tumor cells . Thus - if selectively targeted to the tumor - siRNA-induced inhibition of complement regulation may serve as an innovative strategy to potentiate the efficacy of antibody-based immunotherapy . 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 . 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 . 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 . Neuregulin-1 activates the JAK- P35610 pathway and regulates lung epithelial cell proliferation . Neuregulin-1 ( Q99988 ) is part of a family of proteins whose members are structurally related to epidermal growth factor . Q99988 induces cell proliferation through a high-affinity receptor complex composed of a heterodimer of human epidermal growth factor-like receptor ( HER ) 2 and 3 . In this study , we show that Q99988 activates the Janus kinases ( JAK ) and signal transducer and activator of transcription proteins ( P35610 ) . Q99988 induced a rapid and transient increase in tyrosine phosphorylation of P29597 and P52333 , but not P23458 or O60674 , and induced P40763 and P42229 tyrosine phosphorylation . Upon phosphorylation , P40763 translocated to the nucleus within 1 h . Activation of the JAK- P35610 pathway was dependent on P04626 / P21860 heterodimerization and was necessary for Q99988 -induced proliferation . Inhibition of P04626 's ability to dimerize using the P04626 -specific antibody 2C4 completely blocked Q99988 -induced P52333 , P29597 , P40763 , and P42229 tyrosine phosphorylation . Blocking the JAK- P35610 pathway with a specific JAK- P35610 pathway inhibitor , AG490 , inhibited Q99988 -induced JAK and P35610 phosphorylation and cell proliferation . These data suggest that Q99988 activates the JAK- P35610 signal transduction pathway through its high-affinity receptor , the P04626 / P21860 heterodimer . This pathway plays an important role in Q99988 -stimulated proliferation of pulmonary epithelial cells . 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 . 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 . DB01892 , an Anti-Inflammatory Constituent from St . John 's Wort , Inhibits Microsomal Prostaglandin E(2) Synthase-1 and Suppresses Prostaglandin E(2) Formation in vivo . The acylphloroglucinol hyperforin ( Hyp ) from St . John 's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of P09917 . Here , we investigated whether Hyp also interferes with prostanoid generation in biological systems , particularly with key enzymes participating in prostaglandin (PG)E(2) biosynthesis , i.e. , cyclooxygenases ( P36551 ) -1/2 and microsomal PGE(2) synthase ( mPGES ) -1 which play key roles in inflammation and tumorigenesis . Similar to the mPGES-1 inhibitors MK-886 and MD-52 , Hyp significantly suppressed PGE(2) formation in whole blood assays starting at 0.03-1 μM , whereas the concomitant generation of P36551 -derived 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid , thromboxane B(2) , and 6-keto P49763 (1α) was not significantly suppressed up to 30 μM . In cell-free assays , Hyp efficiently blocked the conversion of PGH(2) to PGE(2) mediated by mPGES-1 ( IC(50) = 1 μM ) , and isolated P36551 enzymes were not ( P35354 ) or hardly ( P23219 ) suppressed . Intraperitoneal ( i.p. ) administration of Hyp ( 4 mg kg(-1) ) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels , and Hyp ( given i.p. ) inhibited carrageenan-induced mouse paw edema formation ( ED(50) = 1 mg kg(-1) ) being superior over indomethacin ( ED(50) = 5 mg kg(-1) ) . We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp . 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 . Mammographic density and candidate gene variants : a twins and sisters study . BACKGROUND : Mammographic density , the light/white radiographic appearance on a mammogram that represents connective and epithelial tissue , is a strong risk factor for breast cancer which seems to be highly heritable . Little is known about its genetic determinants . METHODS : We studied 457 women from 207 sisterhoods ( 104 monozygotic twins , 182 dizygotic twins , and 171 singletons ) . Percentage mammographic density ( PMD ) as well as dense area and nondense area were calculated using a computer-assisted method . We measured six single nucleotide polymorphisms from six candidate genes ( P21964 , P14060 , P17936 , P04626 , P18074 , and O43542 ) . Associations between genotypes and mammographic measures were tested ( a ) cross-sectionally using a multivariate normal model fitted using FISHER that allowed separate correlations for monozygotic , dizygotic , and nontwin pairs and ( b ) within sister pairs using paired t tests . RESULTS : Cross-sectionally , each additional copy of the P14060 DB00174 (367) DB00156 variant allele was associated with lower PMD ( -3.47 % per allele ; SE = 1.65 ; P = 0.035 ) . Within-pair regression estimates confirmed this association . There was no evidence for an association between the mammographic density measures and any of the other variants studied . CONCLUSION : We have replicated an association between a variant in the P14060 gene and PMD , which suggests that P14060 may be genetic determinant of mammographic density . Host factors mediating HIV-1 replication . Human immunodeficiency virus type 1(HIV-1) infection is the leading cause of death worldwide in adults attributable to infectious diseases . Although the majority of infections are in sub-Saharan Africa and Southeast Asia , HIV-1 is also a major health concern in most countries throughout the globe . While current antiretroviral treatments are generally effective , particularly in combination therapy , limitations exist due to drug resistance occurring among the drug classes . Traditionally , HIV-1 drugs have targeted viral proteins , which are mutable targets . As cellular genes mutate relatively infrequently , host proteins may prove to be more durable targets than viral proteins . HIV-1 replication is dependent upon cellular proteins that perform essential roles during the viral life cycle . DB04835 is the first FDA-approved antiretroviral drug to target a cellular factor , HIV-1 coreceptor P51681 , and serves to intercept viral-host protein-protein interactions mediating entry . Recent large-scale siRNA and shRNA screens have revealed over 1000 candidate host factors that potentially support HIV-1 replication , and have implicated new pathways in the viral life cycle . These host proteins and cellular pathways may represent important targets for future therapeutic discoveries . This review discusses critical cellular factors that facilitate the successive steps in HIV-1 replication .

MH_train_1597

MH_train_1597

MH_train_1597

interacts_with DB09068?

multiple_choice

A DNA hypermethylation profile reveals new potential biomarkers for prostate cancer diagnosis and prognosis . BACKGROUND : DNA hypermethylation has emerged as a novel molecular biomarker for the evaluation of prostate cancer diagnosis and prognosis . Defining the specific gene hypermethylation profile for prostate cancer could involve groups of genes that specifically discriminate patients with indolent and aggressive tumors . METHODS : Genome-wide methylation analysis was performed on 83 tumor and 10 normal prostate samples using the GoldenGate Methylation Cancer Panel I ( Illumina , Inc. ) . All clinical stages of disease were considered . RESULTS : We found 41 genes hypermethylated in more than 20 % of the tumors analyzed ( P < 0.01 ) . Of these , we newly identified P28161 and P01210 as being genes that are hypermethylated in prostate cancer and that were simultaneously methylated in 40.9 % of the tumors analyzed . We also identified panels of genes that are more frequently methylated in tumor samples with clinico-pathological indicators of poor prognosis : a high Gleason score , elevated Ki-67 , and advanced disease . Of these , we found simultaneous hypermethylation of P13569 and P28222 to be common in patients with a high Gleason score and high Ki-67 levels ; this might indicate the population at higher risk of therapeutic failure . The DNA hypermethylation profile was associated with cancer-specific mortality ( log-rank test , P = 0.007 ) and biochemical recurrence-free survival ( log-rank test , P = 0.0008 ) . CONCLUSIONS : Our findings strongly indicate that epigenetic silencing of P28161 and P01210 is a common event in prostate cancer that could be used as a molecular marker for prostate cancer diagnosis . In addition , simultaneous P28222 and P13569 hypermethylation could help discriminate aggressive from indolent prostate tumors . 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 . Integrative analysis of miRNA and mRNA expression profiles in pheochromocytoma and paraganglioma identifies genotype-specific markers and potentially regulated pathways . Pheochromocytomas ( PCCs ) and paragangliomas ( PGLs ) are rare neuroendocrine neoplasias of neural crest origin that can be part of several inherited syndromes . Although their mRNA profiles are known to depend on genetic background , a number of questions related to tumor biology and clinical behavior remain unanswered . As microRNAs ( miRNAs ) are key players in the modulation of gene expression , their comprehensive analysis could resolve some of these issues . Through characterization of miRNA profiles in 69 frozen tumors with germline mutations in the genes O14521 , P21912 , P40337 , P07949 , P21359 , O75204 , and P61244 , we identified miRNA signatures specific to , as well as common among , the genetic groups of PCCs/PGLs. miRNA expression profiles were validated in an independent series of 30 composed of P40337 - , P21912 - , O14521 - , and P07949 -related formalin-fixed paraffin-embedded DB09058 /PGL samples using quantitative real-time PCR . Upregulation of miR-210 in P40337 - and P21912 -related PCCs/PGLs was verified , while miR-137 and miR-382 were confirmed as generally upregulated in PCCs/PGLs ( except in P61244 -related tumors ) . Also , we confirmed overexpression of miR-133b as P40337 -specific miRNAs , miR-488 and miR-885-5p as P07949 -specific miRNAs , and miR-183 and miR-96 as P21912 -specific miRNAs . To determine the potential roles miRNAs play in DB09058 /PGL pathogenesis , we performed bioinformatic integration and pathway analysis using matched mRNA profiling data that indicated a common enrichment of pathways associated with neuronal and neuroendocrine-like differentiation . We demonstrated that miR-183 and/or miR-96 impede P01138 -induced differentiation in PC12 cells . Finally , global proteomic analysis in P21912 and P61244 tumors allowed us to determine that miRNA regulation occurs primarily through mRNA degradation in PCCs/PGLs , which partially confirmed our miRNA-mRNA integration results . Detection of K-ras mutation in sputum by mutant-allele-specific amplification ( Q9UHY7 ) . From 10 to 30 % of lung carcinomas examined to date contain mutant K-ras genes . We report here that the mutant-allele-specific amplification ( Q9UHY7 ) method may be useful for detection of the K-ras mutations in cells obtained from the sputum of patients with lung cancer . The PCR product from one of five patients revealed an alteration when mixed oligonucleotides representing variants of the second letter at codon 12 of this gene were used as 5' primers , and further experiments showed a mutation of P19440 ( DB00145 ) to Q6IB77 ( DB00128 ) at codon 12 . The Q9UHY7 system could also be applied to an examination of metastatic lung carcinomas , particularly from adenocarcinomas in colon and pancreas in which frequent K-ras mutations are detected , and to mass-screening for colorectal tumors using DNA isolated from feces as template . DB00939 sodium is an inhibitor of both the P09917 and cyclooxygenase pathways of the arachidonic acid cascade in vitro . DB00939 sodium was compared to other nonsteroidal antiinflammatory drugs in terms of its potency to inhibit the formation of 5-HETE and LTB4 in human leukocytes and the formation of prostaglandin E2 in bovine seminal vesicles as measures of its ability to inhibit the P09917 and cyclooxygenase pathways of the arachidonic acid cascade . DB00939 sodium was about 2-4 times less potent than BW-755C in inhibiting P09917 enzyme activity and three times more potent than benoxaprofen , while naproxen , ibuprofen , and indomethacin showed IC50 greater than 100 microM . DB00939 sodium and indomethacin were the most potent inhibitors of the formation of DB00917 in bovine seminal vesicles followed by ibuprofen , naproxen , and benoxaprofen in this order . DB00939 sodium , like BW-755C , can be considered a dual inhibitor of P09917 and cyclooxygenase pathways of arachidonic acid cascade . This finding may explain in part the antiinflammatory activity of meclofenamate sodium . 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 . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . The P08908 receptor agonist Bay x 3702 inhibits apoptosis induced by serum deprivation in cultured neurons . We examined whether the highly selective P08908 receptor agonist (-)-(R)-2- [ 4- [ [ (3,4-dihydro-2H-1-benzopyran-2-yl)methyl ] -amino ] butyl ] -11 , 2-benz-isothiazol-3(2H)-one 1,1-dioxide monohydrochloride ( Bay x 3702 ) could inhibit neuronal apoptosis induced by serum deprivation . In primary cultures of chick embryonic neurons and in mixed neuronal/glial cultures from neonatal rat hippocampus , Bay x 3702 ( 1 microM ) rescued serum-deprived neurons from apoptosis . The antiapoptotic effect of Bay x 3702 ( 1 microM ) was blocked in chick neurons by the selective P08908 receptor antagonists 4-iodo-N-[2-[4-(methoxyphenyl)-1-piperazin]ethyl]-N-2-pyridinyl-be nzamide hydrochloride ( p-MPPI , 10 microM ) and 4-[3-benzotriazol-1-propyl]-1-(2-methoxyphenyl)-piperazine ( BPMP , 10 microM ) as well as by anti-nerve growth factor ( anti- P01138 ) antibodies and in rat neurons by N-[2-4-(2-methoxy)-1-piperazinyl]ethyl ] -N-(2-pyridinyl)cyclohexane-carbo xamide trihydrochloride ( WAY 100635 , 10 microM ) . We found only under control conditions ( medium with serum ) , but not in serum-deprived cultures , that P01138 secretion was 6-fold increased by Bay x 3702 ( 1 microM ) compared to untreated cultures . Additionally , Bay x 3702 ( 4 microg/kg i.v. ) , infused within a period of 4 h , significantly increased the P01138 content of the rat hippocampus , but not of the striatum . In summary , our data suggest that Bay x 3702 inhibited growth factor withdrawal-induced apoptosis by the stimulation of P08908 receptors and that the P01138 signalling pathway is involved in the mechanism of action . H. pylori activates NF-kappaB through a signaling pathway involving IkappaB kinases , NF-kappaB-inducing kinase , TRAF2 , and Q9Y4K3 in gastric cancer cells . BACKGROUND & AIMS : H. pylori infection on gastric epithelial cells has been shown to induce NF-kappaB activation , but the mechanism of intracellular signal conduction that leads to NF-kappaB activation is not clear . The aim of this study was to analyze the molecular mechanism responsible for H. pylori-mediated NF-kappaB activation on gastric cancer cells . METHODS : NF-kappaB activation by H. pylori was tested by using luciferase reporter assay . P25963 degradation by H. pylori infection was assessed by immunoblotting . IKKalpha and IKKbeta activation was analyzed by kinase assay . In transfection experiments , effects of dominant negative P25963 , IKKalpha , IKKbeta , NF-kappaB-inducing kinase ( NIK ) , TRAF2 , and Q9Y4K3 mutants were investigated . The effects of an IKKbeta-specific inhibitor , aspirin , on NF-kappaB activation and P10145 secretion were also analyzed . RESULTS : H. pylori promotes degradation of P25963 , a cytoplasmic inhibitor of NF-kappaB . In kinase assay , H. pylori induced IKKalpha and IKKbeta catalytic activity in gastric cancer cells . Transfection of kinase-deficient mutant of either IKK inhibited H. pylori-mediated NF-kappaB activation dose-dependently . DB00945 inhibited both NF-kappaB activation and P10145 secretion induced by H. pylori . NF-kappaB activation was also inhibited by transfection of kinase-deficient NIK or a dominant negative mutant of upstream adapter protein TRAF2 or Q9Y4K3 . CONCLUSIONS : H. pylori induces NF-kappaB activation through an intracellular signaling pathway that involves IKKalpha , IKKbeta , NIK , TRAF2 , and Q9Y4K3 . Attenuated P08908 receptor signaling in brains of suicide victims : involvement of adenylyl cyclase , phosphatidylinositol 3-kinase , Akt and mitogen-activated protein kinase . Positron emission tomography studies in major depression show reduced serotonin (5-HT)1A receptor antagonist-binding potentials in many brain regions including occipital cortex . The functional meaning of this observation in terms of signal transduction is unknown . We used postmortem brain samples from depressed suicide victims to examine the downstream effectors of P08908 receptor activation . The diagnosis was established by means of psychological autopsy using Diagnostic and Statistical Manual of Mental Disorders ( DSM ) III-R criteria . Measurements of [35S]GTPgammaS binding to Galphai/o in the occipital cortex of suicide victims and matched controls revealed a blunted response in suicide subjects and a decrease in the coupling of P08908 receptor to adenylyl cyclase . No significant group differences were detected in the expression levels of Galphai/o , Galphaq/11 or Galphas proteins , or in the activity of DB02527 -dependent protein kinase A . Studies of a parallel transduction pathway downstream from P08908 receptor activation demonstrated a decrease in the activity of phosphatidylinositol 3-kinase and its downstream effector Akt , as well as an increase in P60484 ( phosphatase and tensin homolog deleted on chromosome 10 ) , the phosphatase that hydrolyzes phosphatidylinositol 3,4,5-triphosphate . Finally , the activation of extracellular signal-regulated kinases 1 and 2 was attenuated in suicide victims . These data suggest that the alterations in agonist-stimulated P08908 receptor activation in depressed suicide victims are also manifest downstream from the associated G protein , affecting the activity of second messengers in two P08908 receptor transduction pathways that may have implications for cell survival . The role of dopamine-metabolizing enzymes in the regulation of renal sodium excretion in the rat . The intrarenal natriuretic hormone dopamine ( DA ) is metabolized by catechol-O-methyltransferase ( P21964 ) and monoamine oxidase ( MAO ) . We have previously shown that inhibition of P21964 by entacapone results in a potent D1-like receptor-mediated natriuretic response . The present study was performed using anaesthetized rats to compare the importance of MAO and P21964 in DA-mediated natriuresis by use of the MAO inhibitor phenelzine . Urinary sodium and DA excretion remained unchanged after MAO inhibition , while excretion of the main metabolite dihydroxyphenylacetic acid ( DOPAC ) decreased by 55 % . The response was unaltered if 5-hydroxytryptamine receptors ( P08908 ) were blocked during MAO inhibition . We also investigated the specific renal activities of MAO and P21964 in rat renal cortex during DA-influenced natriuresis . Specific P21964 activity in the renal cortex was reduced by 13 % after isotonic sodium loading ( 5 % of body mass ) whereas renal P21397 and P27338 activities remained unaltered . Furthermore , preliminary data obtained from spontaneously hypertensive rats , whose basal urinary DA excretion is higher than that of normotensive Wistar-Kyoto rats , show a tendency for renal P21964 activity to be lower . It is concluded that MAOinhibition by phenelzine does not alter sodium excretion . Furthermore , specific renal cortical P21964 activity is reduced during partly D1-like receptor-mediated natriuresis , whereas MAO activity remains unchanged . The results suggest that MAO is less important than P21964 in regulating DA-mediated natriuresis in the rat kidney . Effect of increased serotonin levels on [18F]MPPF binding in rat brain : fenfluramine vs the combination of citalopram and ketanserin . [18F]MPPF is a selective serotonin-1A ( P08908 ) receptor antagonist and may be used to measure changes in the functional levels of serotonin ( 5-HT ) . The technique is based on the assumption that the injected radiolabeled ligand competes for the same receptor as the endogenous transmitter . Results from studies using serotonergic ligands are not always consistent . The aim of the present study was to investigate if [18F]MPPF binding is decreased after an increase in 5-HT levels . [18F]MPPF binding was assessed in conscious rats using ex vivo autoradiography . We studied the effect of the 5-HT-releasing agent and reuptake inhibitor fenfluramine ( 10 mg/kg i.p. ) and of a combination of the selective serotonin reuptake inhibitor ( SSRI ) citalopram ( 10 micromol/kg , s.c. ) with the P28335 antagonist ketanserin ( 100 nmol/kg , s.c ) . The effect of both treatments on extracellular 5-HT levels was determined using microdialysis . DB00574 treatment resulted in a 30-fold increase in extracellular 5-HT levels in the ventral hippocampus and induced a significant reduction of [18F]MPPF binding in the frontal cortex , hypothalamus , amygdala , and hippocampus . The microdialysis results showed a 10-fold 5-HT increase in the ventral hippocampus after combined administration of ketanserin and citalopram . The combination , however , did not affect [18F]MPPF binding . Our data show that [18F]MPPF binding in conscious rats is only reduced after substantial and therefore nonphysiological increases in 5-HT levels . These results may imply that the majority of P08908 receptors is in the low-affinity state , in vivo . Inhibition of noradrenaline release via presynaptic P28222 receptors of the rat vena cava . In the rat inferior vena cava preincubated with 3H-noradrenaline , the effects of nine serotonin ( 5-HT ) receptor agonists and of eight antagonists ( including two beta-adrenoceptor blocking agents ) on the electrically evoked 3H overflow were determined . 1 . 5-HT , 5-carboxamido-tryptamine , 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole ( RU 24969 ) , 5-methoxytryptamine , N,N-dimethyl-5HT , tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow . The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for P28222 binding sites , but not with their affinities for P08908 , P28335 or 5-HT2 binding sites . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 receptor agonist , and ipsapirone , a partial agonist at these receptors , did not inhibit overflow . 2 . Cyanopindolol facilitated the evoked 3H overflow , an effect which was abolished by propranolol . The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol . 3 . The concentration-response curve for 5-HT was shifted to the right by metitepine , metergoline , quipazine , 6-chloro-2-(1-piperazinyl)pyrazine ( MK 212 ) and propranolol which , given alone , did not affect 3H overflow . The apparent pA2 values of these antagonists tended to be correlated with their affinities for P28222 ( but not P08908 , P28335 or 5-HT2 ) binding sites . Ketanserin , a 5-HT2 receptor antagonist , and spiperone , which blocks 5-HT2 and P08908 but not P28222 or P28335 receptors , failed to antagonize the effect of 5-HT. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . DB00139 dehydrogenase subunit B ( P21912 ) gene deletion associated with a composite paraganglioma/neuroblastoma . 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 . Stress and antidepressant effects on hippocampal and cortical P08908 and 5-HT2 receptors and transport sites for serotonin . The interactions between 14 days of repeated restraint stress and daily administration of imipramine or tianeptine ( 2 h before the beginning of stress ) were investigated in rats to assess responses of 5-HT2 and P08908 receptors and serotonin transporter sites labelled by [3H]paroxetine in the cerebral cortex and hippocampus , two brain regions in which adrenal steroid effects on serotonin receptor-binding have been reported . 5-HT2 sites , labelled by [125I]7-amino-8-iodo ketanserin , were decreased in parietal cerebral cortex layers 3 and 5 by imipramine treatment , but not by tianeptine treatment and not by daily restraint stress . Stress , but not antidepressant , depressed P08908 sites labelled with [3H]8-hydroxy-DPAT in hippocampal fields P07451 , P22748 and dentate gyrus . [3H]paroxetine-binding to serotonin transporter sites was decreased by tianeptine treatment as well as by imipramine in both hippocampus and cerebral cortex , with some overlap of the fields that were significantly affected , whereas there were no effects of stress per se and no evidence of a stress x drug interaction . These results are discussed in relation to similarities and differences in the effects of different antidepressant drugs on the serotonergic system of the rat brain . Whereas the actions of imipramine and tianeptine on 5-HT2 and P08908 receptors are specific to each drug , the surprising finding of a similar effect of both drugs to reduce serotonin transporter sites labelled by [3H]paroxetine suggest the possibility of a common action for these two drugs in spite of their opposite effects on serotonin re-uptake . Inhibition of P08908 receptor-dependent cell survival by DB02527 /protein kinase A : role of protein phosphatase 2A and Bax . Serotonergic 5-HT(1A) receptor signaling leading to nuclear factor-kappaB ( NF-kappaB ) activation appears to be critical for cell survival . Adenylyl cyclase and protein kinase A ( AC/PKA ) are effectors of the 5-HT(1A) receptor that are inhibited by Galpha(i) subunits . Conversely , Gbetagamma(i) subunits downstream from the 5-HT(1A) receptor participate in the activation of extracellular signal-regulated kinases ( P27361 /2 ) , phosphatidylinositol 3-kinase ( PI3K ) , Akt , and NF-kappaB . To model the contribution of pro- and antiapoptotic signaling cascades downstream of activated 5-HT(1A) receptor in cell survival , Chinese hamster ovarian ( CHO ) cells were employed that exogenously overexpress 5-HT(1A) receptors . Stimulation with the 5-HT(1A) receptor agonist 8-OH-DPAT and pharmacological agonists of AC induced PKA and protein phosphatase 2A ( PP2A ) activity , which in turn inhibited : Akt activity , P25963 degradation , nuclear translocation of NF-kappaB , and expression of P98170 ( P98170 / P98170 ) . Pharmacological inhibition of PP2A with calyculin A potentiated Akt activity while attenuating P27361 /2 signaling via increased inhibitory phosphorylation of Raf ( pSer259 ) . In contrast , increased DB02527 levels enhanced Bax translocation to the mitochondria , resulting in the release of cytochrome c , caspase-3 activation , and apoptosis induction . Our data suggest a central role of DB02527 /PKA-dependent PP2A in shifting the homeostasis of intracellular signaling downstream of activated 5-HT(1A) receptor toward cell death in biological systems linked to neuropsychiatric disorders . [ Influence of serotonergic transmission on response to olanzapine ] . INTRODUCTION : This study aimed to investigate associations between the response to olanzapine and genetic variations ( polymorphisms ) in serotonergic transmission related genes in a sample of prospectively studied schizophrenic patients treated with this drug . METHODOLOGY : A total of 51 non-related patients with a DSM-IV diagnosis of schizophrenia were treated with olanzapine ( mean dose : 12 mg/day ; range : 5-25 mg ) and followed-up for at least three months . Response to olanzapine was measured by the difference between baseline and post-treatment scores on the PANSS and GAS scales . The following polymorphisms were studied : serotonin receptor 5- Q13049 ( 102-T/C , His452Tyr ) , serotonin receptor P28335 ( Cys23Ser , -330- P19440 /-244-CT ) , and serotonin transporter ( VNTR , 5-HTTLPR ) . RESULTS : Global clinical improvement , measured with both the GAS and PANSS total scores , was observed . When patients were divided into responders and non-responders , the distribution of genotypic and allelic frequencies was similar to the one observed in previous studies with clozapine . When regression analyses were undertaken , polymorphism 330-GT/-244-CT of the P28335 serotonin receptor and 5-HTTLPR of the serotonin transporter showed a tendency towards the association to olanzapine response . CONCLUSIONS : The present study provides preliminary evidence of the important role of variations in serotonin transmission related genes in determining clinical response to olanzapine . Considering previous studies , it can also be concluded that olanzapine and clozapine may have similar affinities to serotonin receptors . 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 . Novel combination treatments targeting chronic myeloid leukemia stem cells . Chronic myeloid leukemia ( CML ) is currently considered incurable in most patients . Stem cell transplantation , an accepted curative option for which extensive experience has been gained , is limited by high morbidity and mortality rates , particularly in older patients . Tyrosine kinase inhibitors targeting P11274 - P00519 are widely used and induce remission in a high proportion of patients , but resistance and incomplete response to these agents portends eventual relapse and disease progression . Although P11274 - P00519 inhibitors eradicate most CML cells , they are largely ineffective against the reservoir of quiescent leukemic stem cells ( LSCs ) . Thus a strong medical need exists for therapies that effectively eradicate LSCs and is currently a focus of extensive research . To date , evidence obtained from in vitro studies , animal models , and clinical CML specimens suggests that an effective approach may be to partner existing P11274 - P00519 inhibitors with compounds targeting key stem cell molecular effectors , including Wnt/β-catenin , hedgehog pathway components , histone deacetylase ( HDAC ) , transforming growth factor-β ( TGF-β ) , O60674 , promyelocytic leukemia protein , and arachidonate P09917 ( P09917 ) . Novel combinations may sensitize LSCs to P11274 - P00519 inhibitors , thereby overcoming resistance and creating the possibility of improving disease outcome beyond the current standard of care .

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interacts_with DB00682?

multiple_choice

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 . A reporter gene assay for screening of DB05876 subtype selective inhibitors . Phosphodiesterase ( PDE ) constitutes a superfamily of enzymes that catalyze the hydrolysis of DB02527 and cGMP into their corresponding monophosphates and play an important role in diverse physiological functions . The present study provides a process for identifying DB05876 subtypes selective inhibitors using a reporter gene assay . Stable recombinant P29320 -293 cell lines expressing high levels of PDE4A4B , PDE4B2A , and PDE4D3 subtypes individually were generated . Transient transfection of pCRE-Luc plasmid , harboring luciferase reporter gene under the control of DB02527 response element ( CRE ) -binding sequence , into these stable recombinant cell lines followed by treatment with DB05876 inhibitor , resulted in a dose dependent increase in luciferase activity . This methods provide a novel , simple and sensitive assay for high throughput screening of DB05876 subtype selective inhibitors for treatment of asthma and P48444 . DB06699 . DB06699 is a gonadotropin-releasing hormone ( DB00644 ) receptor antagonist that , in common with P30968 agonists ( e.g. leuprolide , goserelin and triptorelin ) , is indicated for use as an androgen-deprivation therapy in patients with advanced prostate cancer . In 1-year , randomized , open-label , phase II or III trials in patients with all stages of prostate cancer , subcutaneous degarelix was associated with rapid , profound and sustained suppression of serum testosterone and prostate-specific antigen ( PSA ) , without evidence of testosterone surges or microsurges . In the phase III trial , degarelix ( 240 mg initially followed by 80 mg every 28 days ) was considered to be effective and noninferior to intramuscular leuprolide ( 7.5 mg every 28 days ) with regard to inducing and maintaining suppression of serum testosterone to castrate levels ( i.e. < or=0.5 ng/mL ) . DB06699 induced testosterone suppression more rapidly than leuprolide . Median serum testosterone levels of < or=0.5 ng/mL were achieved by day 3 in degarelix recipients , but not until day 28 in leuprolide recipients . PSA suppression was also more rapid with degarelix than with leuprolide , with significant between-group differences in serum PSA levels favouring degarelix at 14 and 28 days . DB06699 treatment for 1 year was generally well tolerated ; the adverse events reported were mostly related to subcutaneous drug administration ( i.e. injection-site reactions ) and hormonal androgen deprivation ( e.g. hot flushes ) . Ameliorative Effect of a Selective Endothelin P25101 Receptor Antagonist in Rat Model of L- DB00134 -induced Vascular Dementia . The present study was designed to investigate the efficacy of selective P25101 receptor antagonist , ambrisentan on hyperhomocysteinemia-induced experimental vascular dementia . L-methionine was administered for 8 weeks to induce hyperhomocysteinemia and associated vascular dementia in male rats . DB06403 was administered to L-methionine-treated effect rats for 4 weeks ( starting from 5(th) to 8(th) week of L-methionine treatment ) . On 52(nd) day onward , the animals were exposed to the Morris water maze ( MWM ) for testing their learning and memory abilities . Vascular endothelial function , serum nitrite/nitrate levels , brain thiobarbituric acid reactive species ( TBARS ) , brain reduced glutathione ( DB00143 ) levels , and brain acetylcholinesterase ( P22303 ) activity were also measured . L-methionine-treated animals showed significant learning and memory impairment , endothelial dysfunction , decrease in/serum nitrite/nitrate and brain DB00143 levels along with an increase in brain TBARS levels and P22303 activity . DB06403 significantly improved hyperhomocysteinemia-induced impairment of learning , memory , endothelial dysfunction , and changes in various biochemical parameters . These effects were comparable to that of donepezil serving as positive control . It is concluded that ambrisentan , a selective P25101 receptor antagonist may be considered as a potential pharmacological agent for the management of hyperhomocysteinemia-induced vascular dementia . 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 ) . 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 . 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 . 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 . DB00126 is dispensable for oxygen sensing in vivo . Prolyl-4-hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl-4-hydroxylase domain ( P20941 ) enzymes requires oxygen and 2-oxoglutarate as cosubstrates with iron(II) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo(-/-) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo(-/-) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum- and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF-1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl(2) . A Cys201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione . Shared and unique signals of high-altitude adaptation in geographically distinct Tibetan populations . Recent studies have used a variety of analytical methods to identify genes targeted by selection in high-altitude populations located throughout the Tibetan Plateau . Despite differences in analytic strategies and sample location , hypoxia-related genes , including Q99814 and Q9GZT9 , were identified in multiple studies . By applying the same analytic methods to genome-wide SNP information used in our previous study of a Tibetan population ( n = 31 ) from the township of Maduo , located in the northeastern corner of the Qinghai-Tibetan Plateau ( 4200 m ) , we have identified common targets of natural selection in a second geographically and linguistically distinct Tibetan population ( n = 46 ) in the Tuo Tuo River township ( 4500 m ) . Our analyses provide evidence for natural selection based on iHS and XP-EHH signals in both populations at the p < 0.02 significance level for Q99814 , Q9GZT9 , P30519 , and P05093 and for P30613 , Q30201 , and P68871 and P69892 , which have also been reported in other studies . We highlight differences ( i.e. , stratification and admixture ) in the two distinct Tibetan groups examined here and report selection candidate genes common to both groups . These findings should be considered in the prioritization of selection candidate genes in future genetic studies in Tibet . 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 . 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 . 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 . Increases in stimulated secretion of proinflammatory cytokines by blood monocytes following arousal of negative affect : the role of insulin resistance as moderator . We examined the effect of negative affect on changes in stimulated secretion of cytokines by blood monocytes and determined whether insulin resistance ( IR ) , as indexed by the Homeostasis Model Assessment ( HOMA ) , moderated these associations in 58 healthy men ( aged 18-65 years ) . Blood samples and ratings of negative affect were collected at rest and 15min following subjects ' participation in the Anger Recall Interview ( Q9Y4X5 ) . Assessment of lipopolysaccharide ( LPS ) -stimulated secretion of IL-1beta , P05231 , and P01375 was accomplished by ELISA of supernatant . Regression models controlling for age , body mass index , and race/ethnicity revealed that higher HOMA-IR values were associated with larger stress-induced increases in IL-1beta and P01375 ( p < .05 ) . Furthermore , arousal of negative affect during the Q9Y4X5 was differentially associated with stress-induced changes in stimulated secretion of P01375 and P05231 as a function of HOMA-IR ( p < .05 ) . Increases in stimulated cytokine secretion were associated with arousal of negative affect , but only among men with higher HOMA-IR values . Among men with lower HOMA-IR values , arousal of negative affect was associated with diminished cytokine secretion . Collectively , these data suggest that the HOMA-IR moderates the impact that arousal of negative affect has on the ability of blood monocytes to secrete inflammatory cytokines in response to LPS . Stress-induced increases in cytokine secretion among high HOMA-IR men are consistent with the role of inflammation in cardiovascular disease , hypertension , type 2 diabetes as well as the metabolic syndrome and underscore the relevance of negative affect in the etiology of these inflammatory conditions . 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 . Cytochromes P450 from family 4 are the main omega hydroxylating enzymes in humans : CYP4F3B is the prominent player in PUFA metabolism . Human CYP450 omega-hydroxylases of the CYP4 family are known to convert arachidonic acid ( AA ) to its metabolite 20-hydroxyeicosatetraenoic acid ( 20-HETE ) . This study deals with hydroxylations of four PUFAs , eicosatrienoic acid ( P25101 ) , AA , eicosapentaenoic acid ( EPA ) , and docosahexaenoic acid ( DB01708 ) by either human recombinant CYP4s enzymes or human liver microsomal preparations . CYP4F3A and CYP4F3B were the most efficient omega-hydroxylases of these PUFAs . Moreover , the differences in the number of unsaturations of P25101 , AA , and EPA allowed us to demonstrate a rise in the metabolic rate of hydroxylation when the double bond in 14-15 or 17-18 was missing . With the CYP4F enzymes , the main pathway was always the omega-hydroxylation of PUFAs , whereas it was the (omega-1)-hydroxylation with P04798 , P33261 , and P05181 . Finally , we demonstrated that the omega9 and omega3 PUFAs ( P25101 , EPA , and DB01708 ) could all be used as alternative substrates in AA metabolism by human P78329 and -4F3B . Thus , they decreased the ability of these enzymes to convert AA to 20-HETE . However , although P25101 was the most hydroxylated substrate , EPA and DB01708 were the most potent inhibitors of the conversion of AA to 20-HETE . These findings suggest that some physiological effects of omega3 FAs could partly result from a shift in the generation of active hydroxylated metabolites of AA through a CYP-mediated catalysis . Role of acute-phase proteins in interleukin-1-induced nonspecific resistance to bacterial infections in mice . Treatment with a single low dose ( 80 to 800 ng ) of interleukin-1 ( IL-1 ) 24 h before a lethal bacterial challenge of granulocytopenic and normal mice enhances nonspecific resistance . Since IL-1 induces secretion of acute-phase proteins , liver proteins which possess several detoxifying effects , we investigated the role of these proteins in the IL-1-induced protection . Inhibition of liver protein synthesis with D-galactosamine ( P22466 ) completely inhibited the IL-1-induced synthesis of acute-phase proteins . P22466 pretreatment abolished the protective effect of IL-1 on survival completely ( neutropenic mice infected with Pseudomonas aeruginosa ) or partially ( nonneutropenic mice infected with Klebsiella pneumoniae ) . Pretreatment with P05231 , a cytokine induced by IL-1 , did not reproduce the protection offered after IL-1 pretreatment , nor did it enhance or deteriorate the IL-1-enhanced resistance to infection . A protective effect of IL-1 via effects on glucose homeostasis during the acute-phase response was investigated by comparing plasma glucose levels in IL-1-treated mice and control mice before and during infection . Although glucose levels in IL-1-pretreated mice were somewhat higher in the later stages of infection , no significant differences from levels in control mice were present , and the glucose levels in control-treated animals never fell to hypoglycemic values . We conclude that the IL-1-induced nonspecific resistance is mediated neither by the induction of P05231 nor by the effects of IL-1 on glucose homeostasis . Acute-phase proteins generated after IL-1 pretreatment , however , seem to play a critical role in the IL-1-induced protection to infection . 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 . 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 . P03372 -alpha 36 mediates mitogenic antiestrogen signaling in ER-negative breast cancer cells . It is prevailingly thought that the antiestrogens tamoxifen and ICI 182 , 780 are competitive antagonists of the estrogen-binding site of the estrogen receptor-alpha ( ER-α ) . However , a plethora of evidence demonstrated both antiestrogens exhibit agonist activities in different systems such as activation of the membrane-initiated signaling pathways . The mechanisms by which antiestrogens mediate estrogen-like activities have not been fully established . Previously , a variant of ER-α , EP-α36 , has been cloned and showed to mediate membrane-initiated estrogen and antiestrogen signaling in cells only expressing ER-α36 . Here , we investigated the molecular mechanisms underlying the antiestrogen signaling in ER-negative breast cancer MDA-MB-231 and MDA-MB-436 cells that express high levels of endogenous ER-α36 . We found that the effects of both 4-hydoxytamoxifen ( DB04468 ) and ICI 182 , 780 ( ICI ) exhibited a non-monotonic , or biphasic dose response curve ; antiestrogens at low concentrations , elicited a mitogenic signaling pathway to stimulate cell proliferation while at high concentrations , antiestrogens inhibited cell growth . Antiestrogens at l nM induced the phosphorylation of the Src-Y416 residue , an event to activate Src , while at 5 µM induced Src-Y527 phosphorylation that inactivates Src . Antiestrogens at 1 nM also induced phosphorylation of the MAPK/ P29323 and activated the P12004 D1 promoter activity through the Src/ P00533 / P42229 pathways but not at 5 µM . Knock-down of ER-α36 abrogated the biphasic antiestrogen signaling in these cells . Our results thus indicated that ER-α36 mediates biphasic antiestrogen signaling in the ER-negative breast cancer cells and Src functions as a switch of antiestrogen signaling dependent on concentrations of antiestrogens through the P00533 / P42229 pathway . 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 . 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 . DB04690 sensitivity is mediated by the pleiotropic drug resistance network in yeast . The antineoplastic alkaloid camptothecin interferes with the catalytic cycle of P11387 rendering it a cellular poison . DB04690 stabilizes a covalent enzyme-DNA intermediate that is converted into lethal double strand DNA lesions during S phase of the cell cycle . Yeast SCT1 mutants were isolated in a screen for mutations in genes other than P11387 that result in camptothecin resistance . Here we report SCT1 is allelic to PDR1 and that a DB00156 -879 to DB00134 substitution in the PDR1-101 transcription factor confers multiple drug resistance . PDR1 regulates the expression of several gene products including the DB00171 -binding cassette transmembrane transport proteins PDR5 , YOR1 , and SNQ2 . The PDR1 T879M mutant increased PDR5 transcription compared with wild-type PDR1 strains . Deletion of PDR1 or the downstream effector SNQ2 increased cell sensitivity to camptothecin , whereas deletion of YOR1 or PDR5 had little effect on camptothecin sensitivity . However , the camptothecin resistance accompanying P22466 -promoted overexpression of PDR5 suggests some substrate promiscuity among the DB00171 -binding cassette transporters . These data underscore the role of the pleiotropic drug resistance network in regulating camptothecin toxicity and are consistent with a model of decreased intracellular concentrations of camptothecin resulting from the increased expression of the SNQ2 transporter .

MH_train_1599

MH_train_1599

MH_train_1599

interacts_with DB01211?

multiple_choice

Not all monoclonals are created equal - lessons from failed drug trials in Crohn 's disease . The recent success of the anti-integrin antibody DB09033 can barely conceal the fact that the biologics armamentarium in Crohn 's disease has barely evolved beyond P01375 blockers so far . This contrasts with other immune-related diseases considered mechanistically and genetically closely related , such as psoriasis and rheumatoid arthritis , where approved biologics target a variety of independent biological mechanisms . Several pharmacological assets that entered clinical development have proven ineffective , or less effective than originally anticipated . While blockade of Q16552 and its receptor via DB09029 and Brodalumab , respectively , worsened Crohn 's disease , the beneficial effect of IL-12/23 p40 blockade via Ustekinumab appeared confined to a subpopulation of Crohn 's disease patients who have previously failed on P01375 blockers . Clinical development of the IFNγ blocker DB05111 was stopped despite demonstrating some clinical benefit , while the T cell co-stimulation blocker DB01281 did not exhibit any hint towards efficacy in Crohn 's disease . Here I review results from these individual development programmes , and also reflect on the lack of efficacy of the P01375 blocker DB00005 . I will discuss aspects of individual trials that might have confounded their interpretation and highlight the evolution in primary and secondary endpoints that have contributed to increasing robustness of results obtained in recent years . Finally , I suggest that mechanistic studies in murine genetic models combined with exploratory immunological studies incorporated in early drug development may represent the key for identifying the next generation of successful pharmacological targets in Crohn 's disease . Transcriptional regulation of artemin is related to neurite outgrowth and actin polymerization in mature Q86YR7 neurons . Q5T4W7 is a member of the glial cell line-derived neurotrophic factor ( P39905 ) family of ligands that helps to ensure the survival of sensory neurons . We used an in vitro isolated dorsal root ganglia model to study the effects of artemin on the adult rat neuronal system and investigate differentially regulated genes . We found that 285 genes were differentially transcribed by artemin after 3 h of treatment , including genes related to cell adhesion and actin polymerization . A series of genes involved in the regulation of actin dynamics , including coronin , Myr 5 , P42768 interacting protein , cofilin , drebrin and dynamin were down-regulated by artemin , suggesting that it plays a previously undefined role in the regulation of actin polymerization and synaptic vesicle movement . Q5T4W7 also down-regulated the expression of genes related to cell adhesion and matrix assembly , including biglycan , plectin , nestin , neuronatin and the neuron-glia- P62158 -related cell adhesion molecule , which is functionally relevant to neurite elongation in Q86YR7 neurons . Q5T4W7 resulted in increases in total neurite length and branching of the Q86YR7 neurons . Also artemin caused an increase of synaptic vesicle clustering . Our results showed that the inhibition of DNA methylation suppressed the artemin-dependent neurite growth , suggesting that the genetic regulation could be relevant to neurite elongation in mature Q86YR7 . Cancer cell-associated fatty acid synthase activates endothelial cells and promotes angiogenesis in colorectal cancer . Upregulation of fatty acid synthase ( P49327 ) , a key enzyme of de novo lipogenesis , is associated with metastasis in colorectal cancer ( CRC ) . However , the mechanisms of regulation are unknown . Since angiogenesis is crucial for metastasis , we investigated the role of P49327 in the neovascularization of CRC . The effect of P49327 on tumor vasculature was studied in orthotopic CRCs , the chick embryo chorioallantoic membrane ( P62158 ) and Matrigel plug models using immunohistochemistry , immunofluorescent staining and confocal microscopy . Cell secretion was evaluated by ELISA and antibody arrays . Proliferation , migration and tubulogenesis of endothelial cells ( ECs ) were assessed in CRC-EC coculture models . In this study , we found that stable knockdown of P49327 decreased microvessel density in HT29 and HCT116 orthotopic CRCs and resulted in ' normalization ' of tumor vasculature in both orthotopic and P62158 models . Furthermore , P49327 regulated secretion of pro- and antiangiogenic factors , including vascular endothelial growth factor-A ( P15692 ) . Mechanisms associated with the antiangiogenic activity noted with knockdown of P49327 included : downregulation of P15692 (189) , upregulation of antiangiogenic isoform P15692 (165b) and a decrease in expression and activity of matrix metalloproteinase-9 . Furthermore , conditioned medium from P49327 knockdown CRC cells inhibited activation of vascular endothelial growth factor receptor-2 and its downstream signaling and decreased proliferation , migration and tubulogenesis of ECs as compared with control medium . Together , these results suggest that cancer cell-associated P49327 regulates tumor vasculature through alteration of the profile of secreted angiogenic factors and regulation of their bioavailability . Inhibition of P49327 upstream of P15692 and other angiogenic pathways can be a novel therapeutic strategy to prevent or inhibit metastasis in CRC . 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 . [ 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 . Anti-angiogenic agent ramucirumab : meaningful or marginal ? DB05578 ( IMC-1121B ) targets P35968 . DB05578 is being investigated in many malignancies including gastric cancer . The Phase III trial in patients with advanced breast cancer failed to improve the primary end point The REGARD trial , a Phase III study , in patients with advanced gastric cancer in the second line setting , had a marginal improvement in overall survival but did not achieve the expected hazard ratio target ( of 0.69 ) and the median duration of therapy with ramucirumab was meager 8 weeks ( only 2 weeks longer than the placebo 's ) . Other notable agents in the second line setting are docetaxel and irinotecan . Preliminary results of the RAINBOW trial suggest that ramucirumab may be providing more than marginal advantage . In this review , we briefly summarize the process of angiogenesis and address the emerging cost-benefit issues that surround all newly developed agents including ramucirumab . Peptides from purified soybean beta-conglycinin inhibit fatty acid synthase by interaction with the thioesterase catalytic domain . P49327 ( FAS ) is uniquely expressed at high levels in cancer cells and adipose tissue . The objectives of this study were to identify , purify and validate soy FAS inhibitory peptides and to predict their binding modes . Soy peptides were isolated from hydrolysates of purified beta-conglycinin by co-immunoprecipitation and identified using LC-MS/MS . Three peptides , KNPQLR , EITPEKNPQLR and RKQEEDEDEEQQRE , inhibited FAS . The biological activity of these peptides was confirmed by their inhibitory activity against purified chicken FAS ( IC(50) = 79 , 27 and 16 mum , respectively ) and a high correlation ( r = -0.7 ) with lipid accumulation in 3T3- Q9NUQ9 adipocytes . The FAS inhibitory potency of soy peptides also correlated with their molecular mass , pI value and the number of negatively charged and hydrophilic residues . Molecular modeling predicted that the large FAS inhibitory peptides ( EITPEKNPQLR and RKQEEDEDEEQQRE ) bond to the thioesterase domain of human FAS with lower interaction energies ( -442 and -353 kcal.mol(-1) , respectively ) than classical thioesterase inhibitors ( DB01083 , -91 kcal.mol(-1) and C75 , -51 kcal.mol(-1) ) . Docking studies suggested that soy peptides blocked the active site through interactions within the catalytic triad , the interface cavity and the hydrophobic groove in the human FAS thioesterase domain . FAS thioesterase inhibitory activities displayed by the synthetic soy peptides EITPEKNPQLR and RKQEEDEDEEQQRE ( IC(50) = 10.1 +/- 1.6 and 10.7 +/- 4.4 mum , respectively ) were higher than C75 ( 58.7 mum ) but lower than DB01083 ( 0.9 mum ) . This is the first study to identify FAS inhibitory peptides from purified beta-conglycinin hydrolysates and predict their binding modes at the molecular level , leading to their possible use as nutraceuticals . 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 . 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 . Modulation of fatty acid synthase enzyme activity and expression during hepatitis C virus replication . The hepatitis C virus ( HCV ) induces alterations of host cells to facilitate its life cycle . P49327 ( P49327 ) is a multidomain enzyme that plays a key role in the biosynthesis of fatty acids and is upregulated during HCV infection . Herein , we applied activity-based protein profiling ( P05067 ) that allows for the identification of differentially active enzymes in complex proteomic samples , to study the changes in activity of P49327 during HCV replication . For this purpose , we used an activity-based probe based on the P49327 inhibitor DB01083 , and observed an increase in the activity of P49327 in the presence of a subgenomic and a genomic HCV replicon as well as in chimeric SCID/Alb-uPA mice infected with HCV genotype 1a . To study the molecular basis for this increase in P49327 activity , we overexpressed individual HCV proteins in Huh7 cells and observed increased expression and activity of P49327 in the presence of core and NS4B , as measured by western blots and P05067 , respectively . Triglyceride levels were also elevated in accordance with P49327 expression and activity . Lastly , immunofluorescence and P05067 imaging analyses demonstrated that while the abundance and activity of P49327 increases significantly in the presence of HCV , its localization does not change . Together these data suggest that the HCV-induced production of fatty acids and neutral lipids is provided by an increase in P49327 abundance and activity that is sufficient to allow HCV propagation without transporting P49327 to the replication complexes . Meloxicam inhibits prostaglandin E(2) generation via cyclooxygenase 2 in the inflammatory site but not that via cyclooxygenase 1 in the stomach . We studied the effects of meloxicam on prostanoid levels , both in the inflammatory site in rat carrageenin-induced pleurisy and in the rat stomach injected with 1 mol/l NaCl solution , to clarify the relationship between its low gastric toxicity and its relative cyclooxygenase ( P36551 ) 2 selectivity . NS-398 ( 3 mg/kg ) , a highly selective P35354 inhibitor , and meloxicam ( 3 mg/kg ) exhibited anti-inflammatory effects in the pleurisy model . Prostaglandin ( PG ) E(2) thromboxane ( TX ) B(2) and 6-keto- P49763 (1alpha) were detectable in the inflammatory site . Anti-inflammatory doses of NS-398 and meloxicam each suppressed the intrapleural PGE(2) level at 5 h as potently as piroxicam ( 3 mg/kg ) as aspirin ( 100 mg/kg ) , both of which are nonselective P36551 inhibitors . NS-398 was much less potent than the other three in suppressing the levels of TXB(2) and 6-keto- P49763 (1alpha) . These results suggest that PGE(2) may be produced mainly via P35354 in this model and that meloxicam may inhibit P35354 in the inflammatory site . Piroxicam completely inhibited the increase in gastric PGE(2) induced by administering 1 mol/l NaCl solution into the rat stomach . DB04743 ( 3 mg/kg ) , another selective P35354 inhibitor , however , never affected this increase , suggesting that the gastric PGE(2) may be produced via P23219 . The anti-inflammatory dose of meloxicam caused statistically nonsignificant suppression of the PGE(2) level , by approximately 50 % . These results suggest that the potent anti-inflammatory effect of meloxicam , accompanied with low gastric toxicity , may be related to its relative selectivity for P35354 over P23219 . Identification of an evolutionarily conserved heterotrimeric protein complex involved in protein targeting . In Caenorhabditis elegans , lin-2 , lin-7 , and lin-10 genetically interact to control the trafficking of the Let-23 growth factor receptor to the basolateral surface of body epithelia . The human homologue of the lin-10 gene has recently been identified as a member of the Q02410 gene family . The Q02410 proteins contain one phosphotyrosine binding ( PTB ) and two P78352 .Dlg.ZO-1 ( PDZ ) domains as well as an extended amino terminus . We have previously shown that the PTB domain of X11alpha ( also known as Mint1 ) can bind to the amyloid precursor protein ( P05067 ) in a phosphotyrosine-independent fashion and can markedly inhibit the processing of P05067 to the amyloid beta ( Abeta ) peptide . Here , we report that X11alpha directly binds to the mammalian homologue of Lin-2 ( mLin-2 ) , also known as CASK . This binding is mediated by direct interaction between the P62158 Kinase II ( CKII ) -like domain of mLin-2 and the amino terminus of X11alpha . Furthermore , we can detect direct interactions between mLin-2 and mammalian Lin-7 ( mLin-7 ) . In mouse brain , we have identified a heterotrimeric complex that contains mLin-2 , mLin-7 , and X11alpha and that is likely important for the localization of proteins in polarized cells . This complex may play an important role in the trafficking and processing of P05067 in neurons . Inhibition of endocannabinoid neuronal uptake and hydrolysis as strategies for developing anxiolytic drugs . The endocannabinoid system comprises the P21554 and CB2 receptors ( the targets of the Cannabis sativa compound DB00470 ) , the endogenous ligands ( endocannabinoids ) arachidonoyl ethanolamide ( anandamide ) and 2-arachidonoyl glycerol , their synthesizing machinery and membrane transport system , and the hydrolyzing enzymes fatty acid amide hydrolase ( FAAH ) and monoacylglycerol lipase ( Q99685 ) , respectively . The endocannabinoids may act on demand to confer protection against aversive stimuli , which suggests that increasing their brain levels may represent an approach for treatment of anxiety-related disorders . Thus , this article reviews the profile of endocannabinoid reuptake and hydrolysis inhibitors in experimental tests predictive of anxiolytic activity . The FAAH inhibitors and the blockers of anandamide transport , in contrast to direct P21554 receptor agonists , induce anxiolytic effects at doses that do not interfere with motor activity . Q99685 inhibitors also reduce anxiety-like behavior , although they are more likely to impair motor activity . Regarding their mechanisms , increasing anandamide levels induce responses mediated by the P21554 receptor and occluded by the transient receptor potential vanilloid type-1 channels , whereas the effects of increasing 2-arachidonoyl glycerol depend on both P21554 and CB2 receptors . Their neuroanatomical targets include various structures related to anxiety and fear responses . Understanding the pharmacological properties of FAAH and Q99685 inhibitors may contribute toward the development of new anxiolytic interventions based on the endocannabinoid system . Structural basis for selective inhibition of P35354 by nimesulide . DB04743 1 is a novel nonsteroidal antiinflammatory drug which inhibits the enzyme cyclooxygenase 2 ( P35354 ) more selectively than cyclooxygenase 1 ( P23219 ) . Molecular modelling studies have been carried out on complexes of 1 with P23219 and with mutants of P23219 simulating P35354 . These indicate that the mutations I523V and S516A largely contribute to the selectivity . A comparative study with SC-558 2 has also been performed . DB08890 - a secretagogue and antihyperalgesic agent - what next ? Ongoing clinical trials suggest that linaclotide , a first-in-class , 14-amino acid peptide guanylate cyclase-C ( P25092 ) receptor agonist and intestinal secretagogue is an effective treatment for chronic constipation . A study in this issue of the Journal suggests that linaclotide also has antihyperalgesic effects in three common rat models of inflammation- and stress-induced hypersensitivity ( i.e. , acute trinitrobenzene sulfonic acid colitis , water avoidance stress [ P42768 ] , and restraint-induced stress ) but not in naïve animals . In mice , linaclotide at least partly reduces hyperalgesia via P25092 receptors . Dose-effect relationships of linaclotide were complicated and non-linear . This viewpoint discusses human clinical trials with linaclotide and the results of this study . Potential mechanisms and clinical significance of these findings are explored . Collectively , these data suggest that P25092 receptors exert other , as yet poorly understood , effects on gastrointestinal sensitivity in conditions associated with inflammation and/or stress-induced increased intestinal permeability . However , the data need to be confirmed in humans and in long-term animal models . Further studies are also necessary to elucidate the mechanisms as these effects can not be explained by linaclotide 's known effects on epithelial P25092 receptors . 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 . Inhibition of drug-resistant mutants of P00519 , P10721 , and P01133 receptor kinases . To realize the full potential of targeted protein kinase inhibitors for the treatment of cancer , it is important to address the emergence of drug resistance in treated patients . Mutant forms of P11274 - P00519 , P10721 , and the P01133 receptor ( P00533 ) have been found that confer resistance to the drugs imatinib , gefitinib , and erlotinib . The mutations weaken or prevent drug binding , and interestingly , one of the most common sites of mutation in all three kinases is a highly conserved " gatekeeper " threonine residue near the kinase active site . We have identified existing clinical compounds that bind and inhibit drug-resistant mutant variants of P00519 , P10721 , and P00533 . We found that the Aurora kinase inhibitor VX-680 and the p38 inhibitor BIRB-796 inhibit the imatinib- and BMS-354825-resistant P00519 (T315I) kinase . The P10721 / P36888 inhibitor SU-11248 potently inhibits the imatinib-resistant P10721 (V559D/T670I) kinase , consistent with the clinical efficacy of SU-11248 against imatinib-resistant gastrointestinal tumors , and the P00533 inhibitors Q9Y259 -569 and DB05424 , but not GW-572016 and ZD-6474 , potently inhibit the gefitinib- and erlotinib-resistant P00533 (L858R/T790M) kinase . Q9Y259 -569 and DB05424 are already in clinical trials , and our results suggest that they should be considered for testing in the treatment of gefitinib/erlotinib-resistant non-small cell lung cancer . The results highlight the strategy of screening existing clinical compounds against newly identified drug-resistant mutant variants to find compounds that may serve as starting points for the development of next-generation drugs , or that could be used directly to treat patients that have acquired resistance to first-generation targeted therapy . 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 . 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 . 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 . Breast cancer cells can switch between estrogen receptor alpha and ErbB signaling and combined treatment against both signaling pathways postpones development of resistance . The majority of breast cancers are estrogen responsive , but upon progression of disease other growth promoting pathways are activated , e.g. , the ErbB receptor system . The present study focuses on resistance to the pure estrogen antagonist fulvestrant and strategies to treat resistant cells or even circumvent development of resistance . Limited effects were observed when targeting P00533 and ErbB2 with the monoclonal antibodies cetuximab , trastuzumab , and pertuzumab , whereas the pan-ErbB inhibitor DB05424 selectively inhibited growth of fulvestrant resistant cell lines . DB05424 inhibited Erk but not Akt signaling , which as well as Erk is important for antiestrogen resistant cell growth . Accordingly , combination therapy with DB05424 and the Akt inhibitor SH-6 or the Protein Kinase C inhibitor RO-32-0432 was applied and found superior to single agent treatment . Further , the resistant cell lines were more sensitive to DB05424 treatment when grown in the presence of fulvestrant , as withdrawal of fulvestrant restored signaling through the estrogen receptor alpha ( ERalpha ) , partly overcoming the growth inhibitory effects of DB05424 . Thus , the resistant cells could switch between ERalpha and ErbB signaling for growth promotion . Although parental MCF-7 cell growth primarily depends on ERalpha signaling , a heregulin-1beta induced switch to ErbB signaling rescued MCF-7 cells from the growth inhibition exerted by fulvestrant-mediated blockade of ERalpha signaling . This interplay between ERalpha and ErbB signaling could be abrogated by combined therapy targeting both receptor systems . Thus , the present study indicates that upon development of antiestrogen resistance , antiestrogen treatment should be continued in combination with signal transduction inhibitors . Further , upfront combination of endocrine therapy with pan-ErbB inhibition may postpone or even prevent development of treatment resistance . 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 . 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 . DB02342 in the human corpus luteum throughout the luteal phase and its influence on lutein cell steroidogenesis and angiogenic activity . OBJECTIVE : To quantitate 2-methoxyestradiol ( 2-ME ) in human corpus luteum ( CL ) of different ages and to determine the expression of cytochrome-P450-1A1 ( P04798 ) and catechol-O-methyl transferase ( P21964 ) in CL and the action of 2-ME on P , vascular endothelial growth factor ( P15692 ) secretion , and luteal angiogenesis . DESIGN : Experimental study . SETTING : University division of reproductive endocrinology . PATIENT(S) : Twenty-four women of reproductive age . INTERVENTION(S) : CL was collected from 15 women during the minilaparotomy for tubal sterilization . Granulosa lutein cells were harvested 36 hours after hCG administration in patients undergoing IVF . MAIN OUTCOMES MEASURE(S) : Levels of 2-ME were determined by high-performance liquid chromatography in CL . P04798 and P21964 were assessed by immunohistochemistry and Western blot . P and P15692 were measured by radioimmunoassay and ELISA . The angiogenic potential was analyzed using EA.hy926 cells . RESULT(S) : Plasma levels of E₂ decreased in the late luteal phase in association with an increase in luteal tissue of 2-ME concentrations . Concomitantly , there was a significant reduction of angiogenic activity in late CL . There was no significant variation in P04798 and P21964 expression in all CL . In physiological doses , 2-ME inhibited basal P15692 by granulosa lutein cells and diminished the angiogenic activity in conditioned media but did not prevent P and P15692 production stimulated by hCG . CONCLUSION(S) : These data suggest the participation of 2-ME in physiological luteolysis by reducing angiogenesis . However , 2-ME did not prevent in vitro hCG stimulation of P biosynthesis , providing a mechanism for CL rescue in the cycle of conception . Activation of large-conductance , Ca2+-activated K+ channels by cannabinoids . We have examined the effects of the cannabinoid anandamide ( AEA ) and its stable analog , methanandamide ( methAEA ) , on large-conductance , Ca2+-activated K+ ( BK ) channels using human embryonic kidney ( P29320 ) -293 cells , in which the alpha-subunit of the Q12791 ( BK-alpha ) , both alpha- and beta1-subunits ( BK-alphabeta1 ) , or both alpha- and beta4-subunits ( BK-alphabeta4 ) were heterologously expressed . In a whole cell voltage-clamp configuration , each cannabinoid activated BK-alphabeta1 within a similar concentration range . Because methAEA could potentiate BK-alpha , BK-alphabeta1 , and BK-alphabeta4 with similar efficacy , the beta-subunits may not be involved at the site of action for cannabinoids . Under cell-attached patch-clamp conditions , application of methAEA to the bathing solution increased Q12791 activity ; however , methAEA did not alter channel activity in the excised inside-out patch mode even when DB00171 was present on the cytoplasmic side of the membrane . Application of methAEA to P29320 -BK-alpha and P29320 -BK-alphabeta1 did not change intracellular Ca2+ concentration . Moreover , methAEA-induced potentiation of Q12791 currents was not affected by pretreatment with a P21554 antagonist ( AM251 ) , modulators of G proteins ( cholera and pertussis toxins ) or by application of a selective CB2 agonist ( JWH133 ) . Inhibitors of P62158 , PKG , and MAPKs ( W7 , KT5823 , and PD-98059 ) did not affect the potentiation . Application of methAEA to mouse aortic myocytes significantly increased Q12791 currents . This study provides the first direct evidence that unknown factors in the cytoplasm mediate the ability of endogenous cannabinoids to activate Q12791 currents . Cannabinoids may be hyperpolarizing factors in cells , such as arterial myocytes , in which BK channels are highly expressed . DB05578 : preclinical research and clinical development . DB05578 ( IMC-1121B , LY3009806 ) , a fully humanized monoclonal antibody directed against the extracellular domain of vascular endothelial growth factor receptor 2 ( P35968 ) , is a new therapeutic option that selectively inhibits the human P35968 with a much greater affinity than its natural ligands . Based on the promising results of both preclinical and early clinical studies , ramucirumab has been tested in different tumor types either alone or in combination with chemotherapy . While it has recently been granted its first US Food and Drug Administration approval for use as a single agent in patients with advanced or metastatic gastric cancer or gastroesophageal junction carcinoma , its role for metastatic breast cancer or advanced non-small-cell lung cancer is still debated . The aims of this review are to recall and discuss the most significant preclinical and clinical studies that led to the development of ramucirumab and to present the results of the randomized clinical trials that have tested its efficacy in different malignancies , including gastric and lung cancer . DB00171 -sensitive potassium channel activation induces angiogenesis in vitro and in vivo . Intense research is conducted to identify new molecular mechanisms of angiogenesis . Previous studies have shown that the angiogenic effects of hydrogen sulfide ( H2S ) depend on the activation of DB00171 -sensitive potassium channels ( KATP ) and that P23582 ( P09543 ) , which can act through KATP , promotes endothelial cell growth . We therefore investigated whether direct KATP activation induces angiogenic responses and whether it is required for the endothelial responses to P09543 or vascular endothelial growth factor ( P15692 ) . Chick chorioallantoic membrane ( P62158 ) angiogenesis was similarly enhanced by the direct KATP channel activator 2-nicotinamidoethyl acetate ( SG-209 ) and by P09543 or P15692 . The KATP inhibitors glibenclamide and 5-hydroxydecanoate ( 5-HD ) reduced basal and abolished P09543 -induced P62158 angiogenesis . In vitro , the direct KATP openers nicorandil and SG-209 and the polypeptides P15692 and P09543 increased proliferation and migration in bEnd.3 mouse endothelial cells . In addition , P15692 and P09543 induced cord-like formation on Matrigel by human umbilical vein endothelial cells ( HUVECs ) . All these in vitro endothelial responses were effectively abrogated by glibenclamide or 5-HD . In HUVECs , a small-interfering RNA-mediated decrease in the expression of the inwardly rectifying potassium channel ( Kir ) 6.1 subunit impaired cell migration and network morphogenesis in response to either SG-209 or P09543 . We conclude that 1 ) direct pharmacologic activation of KATP induces angiogenic effects in vitro and in vivo , 2 ) angiogenic responses to P09543 and P15692 depend on KATP activation and require the expression of the Kir6.1 KATP subunit , and 3 ) KATP activation may underpin angiogenesis to a variety of vasoactive stimuli , including H2S , P15692 , and P09543 . 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 . Inhibition of Q16552 as a pharmacological approach for Q9UKU7 . Several experimental approaches have been utilized , in order to critically examine the roles of Q16552 family members in intestinal inflammation . These approaches have included : ( 1 ) the use of Q16552 and Q96PD4 -deficient mice , ( 2 ) specific antibodies directed against Q16552 , ( 3 ) an Q16552 vaccine , ( 4 ) methods to block the Q16552 receptor and ( 5 ) small-molecule inhibitors of Q16552 . Previous studies found somewhat conflicting results in preclinical models of Inflammatory Bowel Disease ( Q9UKU7 ) , using specific strains of Q16552 -deficient mice . This paper will review the preclinical results using various pharmacological approaches [ specific Q16552 antibodies , an Q16552 receptor fusion protein , IL-12/IL-23 p40 subunit and Q16552 vaccine approaches , as well as a small molecule inhibitor ( Vidofludimus ) ] to inhibit Q16552 in animal models of Q9UKU7 . Recent clinical results in patients with Q9UKU7 will also be discussed for DB09029 ( an Q16552 antibody ) , Brodalumab ( an Q16552 receptor antibody ) and two small-molecule drugs ( Vidofludimus and DB08895 ) , which inhibit Q16552 as part of their overall pharmacological profiles . This review paper will also discuss some pharmacological lessons learned from the preclinical and clinical studies with anti- Q16552 drugs , as related to drug pharmacodynamics , Q16552 receptor subtypes and other pertinent factors . Finally , future pharmacological approaches of interest will be discussed , such as : ( 1 ) Retinoic acid receptor-related orphan nuclear receptor gamma t ( Rorγt ) antagonists , ( 2 ) P10276 ( RARα ) antagonists , ( 3 ) Pim-1 kinase inhibitors and ( 4 ) Dual small-molecule inhibitors of NF-κB and P40763 , like synthetic triterpenoids . 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 . 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 .