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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 . Introduction to the use of belatacept : a fusion protein for the prevention of posttransplant kidney rejection . The development of new immunosuppressive drugs for kidney transplantation resulted both in better short-term outcomes and in decreased metabolic , cardiovascular , and nephrotoxicity risk . DB06681 belongs to a new class of immunosuppressive drugs that selectively inhibits T-cell activation by preventing P10747 activation and by binding its ligands P33681 -1 and P33681 -2 . The result is an inactivation of costimulatory pathways . A comparative analysis of the BENEFIT and BENEFIT-EXT datasets showed belatacept regimens resulted in better cardiovascular and metabolic risk profiles than did cyclosporin A ( DB00091 ) regimens : belatacept likewise outperformed DB00091 in terms of lower blood pressure and serum lipids and less new onset diabetes after transplantation . About 20 % of belatacept-treated patients developed adverse effects which included anemia , pyrexia , neutropenia , diarrhea , urinary tract infection , headache , and peripheral edema . At present , belatacept does not seem to predispose patients to a higher rate of infection than DB00091 maintenance immunosuppression . The risk of posttransplant lymphoproliferative diseases was higher in Epstein-Barr virus ( EBV ) -seronegative patients than in EBV-seropositive patients , but the risk may be reduced by use of a less intensive regimen and avoidance of EBV-negative patients and of patients whose pretransplant EBV serology is unknown . DB06681 provides a new option for immunosuppressive therapy in kidney transplantation , but needs further evaluation in terms of the late effects that may derive from prolonged blockage of the costimulatory system and the induction of tolerance status . The cloning and reintroduction into animal cells of a functional CAD gene , a dominant amplifiable genetic marker . Rodent cells resistant to DB03459 , a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional P27708 , overproduce CAD as a result of amplification of the CAD gene . We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector . Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long . We introduced the cloned genes into CAD-deficient Chinese hamster ovary ( CHO ) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids . We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of DB03459 following protoplast fusion . The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes . The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization . In two of these transformants , the genes are located in one X chromosome or in a chromosome resembling the X . In the third case , the genes are located in a small metacentric or rearranged chromosome . 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 . P10275 -mediated antagonism of estrogen-dependent low density lipoprotein receptor transcription in cultured hepatocytes . Postmenopausal women receiving hormone replacement therapy have a lower risk of coronary heart disease than women who do not receive hormone treatment . Multiple mechanisms are likely to underlie estrogen 's cardioprotective action , including lowering of plasma low density lipoprotein ( LDL ) cholesterol . Using an in vitro system exhibiting normal regulation of P01130 ( P01130 ) gene transcription , we show that 17beta-estradiol activates the P01130 promoter in transiently transfected HepG2 cells . P01130 activation by estrogen in HepG2 cells is dependent on the presence of exogenous estrogen receptor , and the estrogen-responsive region of the P01130 promoter colocalizes with the sterol response element previously identified . The estrogen response is concentration dependent , saturable , and sensitive to antagonism by estrogen receptor antagonists . Further , we show that compounds with androgen receptor agonist activity attenuate the estrogen-induced up-regulation of P01130 in our model system . Progestins with androgen receptor agonist activity , such as medroxyprogesterone acetate , also suppress estrogen 's effects on P01130 expression through their androgenic properties . Characterization of the interplay between these hormone receptors on the P01130 in vitro system may allow a better understanding of the actions of sex steroids on P01130 gene expression and their roles in cardiovascular disease . DNA methylation regulates adenosine A(2A) receptor cell surface expression levels . DB00640 A(2A) receptors ( A(2A)Rs ) appear to play important roles in inflammation and in certain diseases of the nervous system . Pharmacological modulation of A(2A)Rs is particularly useful in Parkinson 's disease and has been tested in schizophrenia . However , little is known about the regulation of A(2A)R gene ( P29274 ) . A bioinformatic analysis revealed the presence of three CpG islands in the 5' UTR region of human P29274 . Next , HeLa , SH-SY5Y and U87-MG cells were treated for 48 h with 5 muM 5-azacytidine ( Aza ) . Increased A(2A)R levels were demonstrated in HeLa and SH-SY5Y cells when compared with non-treated cells . No modifications were seen in U87-MG cells . The increased A(2A)R mRNA and protein levels were accompanied by a loss of DNA methylation pattern in HeLa and SH-SY5Y cells , as measured with the SEQUENOM MassArray platform . The Aza treatment also reduced the affinity of a methyl- Q9P0U4 for P29274 by quantitative chromatin immunoprecipitation in HeLa cells . Interestingly , A(2A)R levels were reduced by S-adenosyl-l-methionine treatment in U87-MG and methyl- Q9P0U4 affinity was increased for P29274 by quantitative chromatin immunoprecipitation . Therefore , these results show for the first time that DNA methylation plays a role in P29274 transcription and , subsequently , in constitutive A(2A)R cell surface levels . 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 . Traps to catch unwary oncogenes . The MYC proto-oncogene has long been implicated in the control of normal cell growth and its deregulation is associated with the development of neoplasia . The MYC protein has a well-established role as a component of signal-transduction pathways promoting both proliferation and apoptosis . Because signalling pathways that drive cell death and cell proliferation are so tightly coupled , a synergy between genetic lesions leading to suppression of cell death and those promoting cell proliferation is observed during carcinogenesis . We discuss such synergy with respect to the cooperating oncogenes MYC , DB01367 and P10415 . Androgens induce prostate cancer cell proliferation through mammalian target of rapamycin activation and post-transcriptional increases in cyclin D proteins . P10275 ( AR ) plays a central role in prostate cancer , with most tumors responding to androgen deprivation therapies , but the molecular basis for this androgen dependence has not been determined . Androgen [ 5alpha-dihydrotestosterone ( DB02901 ) ] stimulation of LNCaP prostate cancer cells , which have constitutive phosphatidylinositol 3-kinase ( PI3K ) /Akt pathway activation due to P60484 loss , caused increased expression of cyclin D1 , D2 , and D3 proteins , retinoblastoma protein hyperphosphorylation , and cell cycle progression . However , cyclin D1 and D2 message levels were unchanged , indicating that the increases in cyclin D proteins were mediated by a post-transcriptional mechanism . This mechanism was identified as mammalian target of rapamycin ( P42345 ) activation . DB02901 treatment increased P42345 activity as assessed by phosphorylation of the downstream targets P08133 S6 kinase and Q13541 , and P42345 inhibition with rapamycin blocked the DB02901 -stimulated increase in cyclin D proteins . Significantly , DB02901 stimulation of P42345 was not mediated through activation of the PI3K/Akt or mitogen-activated protein kinase/p90 ribosomal S6 kinase pathways and subsequent tuberous sclerosis complex 2/tuberin inactivation or by suppression of AMP-activated protein kinase . In contrast , P42345 activation by DB02901 was dependent on AR-stimulated mRNA synthesis . Oligonucleotide microarrays showed that DB02901 -stimulated rapid increases in multiple genes that regulate nutrient availability , including transporters for amino acids and other organic ions . These results indicate that a critical function of AR in P60484 -deficient prostate cancer cells is to support the pathologic activation of P42345 , possibly by increasing the expression of proteins that enhance nutrient availability and thereby prevent feedback inhibition of P42345 . 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 . 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 ) . P10275 -induced tumor suppressor , B2CW77 , inhibits breast cancer growth and transcriptionally activates p53/p73-mediated apoptosis in breast carcinomas . P10275 ( AR ) expression by immunohistochemistry correlates with better prognosis and survival among breast cancer patients . We and others have shown that AR inhibits proliferation and induces apoptosis in breast cancer cells . However , the mechanism of AR 's anti-tumor effect in breast cancer is still not fully understood . Our recent study indicates that AR upregulates expression of tumor suppressor gene P60484 by promoter activation in breast cancer . B2CW77 , encoding B2CW77 protein , is a newly identified gene , which shares a bidirectional promoter with P60484 and is transcribed in the opposite direction . So far , the function of B2CW77 has never been studied in tumorigenesis . Here , we define B2CW77 as a tumor suppressor in breast carcinomas , which inhibits tumor growth and invasiveness . After analyzing 188 normal breast and 1247 malignant breast cancer tissues , we observed the loss of B2CW77 in multiple breast cancer subtypes and this decreased B2CW77 expression associates with tumor progression and increasing histological grade in invasive carcinomas . We characterize B2CW77 , for the first time , as a transcription factor , directly promoting the expression of P04637 and O15350 , with consequent elevated apoptosis and cell cycle arrest in breast cancer cells . We demonstrate , in vitro and in murine xenograph models , that both B2CW77 and P60484 are AR-target genes , mediating androgen-induced growth inhibition and apoptosis in breast cancer cells . Our observations suggest that B2CW77 might be used as a potential prognostic marker and novel therapy target for breast carcinomas . 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 . 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 . A 77-base pair LINE-like sequence elicits androgen-dependent mvdp/akr1-b7 expression in mouse vas deferens , but is dispensable for adrenal expression in rats . Mvdp/akr1-b7 ( mouse vas deferens protein/aldo-keto reductase 1- P33681 ) encodes an enzyme responsible for detoxification of a steroidogenesis byproduct . MVDP/AKR1- P33681 is expressed in both rat and mouse adrenal cortex under DB01285 control , whereas strong androgen-dependent accumulation in the vas deferens is mouse specific . Comparison of the regulatory regions of the two orthologs reveals a strong identity , disrupted by acquisition of a 77-bp LINE-derived sequence in the mouse promoter . Although DB01285 responsiveness is observed in both species , the absence of this 77-bp sequence in the rat is associated with changes in transcription initiation sites . Transfection studies demonstrate that the CCAAT/enhancer-binding protein and selective promoter factor 1-binding sites previously shown to be essential for DB02527 / DB01285 induction in the mouse are consequently dispensable in the rat . Our data support the idea that the most striking change generated by this acquisition is the strong , androgen-dependent , vas deferens expression observed in mouse . 1 ) In rat vas deferens , rakr1-b7 expression is barely detectable and is not androgen sensitive . 2 ) P10275 binds efficiently to an androgen response element within the 77-bp mouse-specific element . 3 ) Its insertion confers androgen sensitiveness to rakr1-b7 regulatory regions in an androgen response element-dependent manner in transient transfections . We propose that this acquired androgen-responsive region may be responsible for vas deferens androgen-regulated gene expression in vivo . Implicating P21583 complexes in organogenesis in Caenorhabditis elegans . Development of the Caenorhabditis elegans foregut ( pharynx ) is regulated by a network of proteins that includes the Retinoblastoma protein ( P06400 ) ortholog LIN-35 ; the ubiquitin pathway components P0CG48 -18 and Q9Y4X5 ; and PHA-1 , a cytoplasmic protein . Loss of pha-1 activity impairs pharyngeal development and body morphogenesis , leading to embryonic arrest . We have used a genetic suppressor approach to dissect this complex pathway . The lethality of pha-1 mutants is suppressed by loss-of-function mutations in sup-35/ztf-21 and sup-37/ztf-12 , which encode Zn-finger proteins , and by mutations in sup-36 . Here we show that sup-36 encodes a divergent Skp1 family member that binds to several F-box proteins and the microtubule-associated protein Q02083 -1/τ . Like P60880 -35 , P60880 -36 levels were negatively regulated by P0CG48 -18- Q9Y4X5 . We also found that P60880 -35 and P60880 -37 physically associated and that P60880 -35 could bind microtubules . Thus , P60880 -35 , P60880 -36 , and P60880 -37 may function within a pathway or complex that includes cytoskeletal components . Additionally , P60880 -36 may regulate the subcellular localization of P60880 -35 during embryogenesis . We carried out a genome-wide RNAi screen to identify additional regulators of this network and identified 39 genes , most of which are associated with transcriptional regulation . Twenty-three of these genes acted via the LIN-35 pathway . In addition , several S-phase kinase-associated protein (Skp)1-Cullin-F-Box ( P21583 ) components were identified , further implicating P21583 complexes as part of the greater network controlling pharyngeal development . 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 . 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 . 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 . 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 . The molecular mechanisms underlying the reduction of LDL apoB-100 by ezetimibe plus simvastatin . The combination of ezetimibe , an inhibitor of Niemann-Pick C1-like 1 protein ( Q9UHC9 ) , and an P04035 inhibitor decreases cholesterol absorption and synthesis . In clinical trials , ezetimibe plus simvastatin produces greater LDL-cholesterol reductions than does monotherapy . The molecular mechanism for this enhanced efficacy has not been defined . P04114 ( apoB-100 ) kinetics were determined in miniature pigs treated with ezetimibe ( 0.1 mg/kg/day ) , ezetimibe plus simvastatin ( 10 mg/kg/day ) , or placebo ( n = 7/group ) . DB00973 decreased cholesterol absorption ( -79 % ) and plasma phytosterols ( -91 % ) , which were not affected further by simvastatin . DB00973 increased plasma lathosterol ( +65 % ) , which was prevented by addition of simvastatin . The combination decreased total cholesterol ( -35 % ) and LDL-cholesterol ( -47 % ) . VLDL apoB pool size decreased 26 % , due to a 35 % decrease in VLDL apoB production . LDL apoB pool size decreased 34 % due to an 81 % increase in the fractional catabolic rate , both of which were significantly greater than monotherapy . Combination treatment decreased hepatic microsomal cholesterol ( -29 % ) and cholesteryl ester ( -65 % ) and increased P01130 ( P01130 ) expression by 240 % . The combination increased Q9UHC9 expression in liver and intestine , consistent with increased Q12772 expression . DB00973 plus simvastatin decreases VLDL and LDL apoB-100 concentrations through reduced VLDL production and upregulation of P01130 -mediated LDL clearance . miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells . To explore the mechanism of apoptosis induced by cisplatin , the expression of microRNAs ( miRNAs ) and regulating genes in K562 cells was analyzed using reverse transcription PCR , quantitative real-time PCR and enzyme-linked immunosorbent assays . Our results showed that miR-16 , miR-34a-c , miR-17-5p and miR-125 were up-regulated , and their associated oncogenes ( P10415 , Q01094 and O00716 , respectively ) were down-regulated after cisplatin treatment . We also showed that miR-106 and miR-150 were down-regulated while their target genes ( P06400 and P04637 , respectively ) were up-regulated after cisplatin treatment . Moreover , miR-16 , miR-34a-c and miR-17-5p proved to be upstream factors , regulating the expression of P10415 , Q01094 and O00716 , respectively . The oncogene O00716 was down-regulated when P06400 expression was increased after treatment with antisense oligonucleotides ( ASO ) . Similarly , P10415 and O00716 were down-regulated when P04637 expression was elevated by ASO treatment . The study demonstrated that cisplatin induces K562 cells to apoptosis by reducing miR-106 which up-regulates P06400 or by inhibiting miR-150 which increases P04637 expression .

MH_train_1401

MH_train_1401

MH_train_1401

interacts_with DB01076?

multiple_choice

P37231 and aging : one link through klotho ? P37231 , a transcription factor involved in adipogenesis , glucose homeostasis , bone turnover , and inflammation , has now been shown to increase klotho expression . Q9UEF7 is predominantly expressed by the kidney and functions at the tubule and in the circulation as an anti-aging factor and a hormone participating in mineral metabolism . The finding that klotho is upregulated by P37231 may prompt further exploration of the role and mechanism of action of P37231 in aging and bone diseases . DB04958 for systemic lupus erythematosus . DB04958 ( EMab , UCB , Immunomedics ) is a humanized monoclonal antibody targeting P20273 that is being studied in clinical trials for patients with a variety of rheumatic and hematologic conditions , including systemic lupus erythematosus ( SLE ) . An overview of its mechanism of action is followed by a summary of completed lupus studies , and a preview of studies in progress . The agent clearly has anti-inflammatory activity and is a potentially useful agent in the management of autoimmune disorders . Inhibitory effects of vinpocetine on the progression of atherosclerosis are mediated by Akt/NF-κB dependent mechanisms in apoE-/- mice . BACKGROUND : Recent studies have found additional roles for vinpocetine , a potent phosphodiesterase type I inhibitor , in anti-proliferation and anti-inflammation of vascular smooth muscle cells and cancer cells via different mechanisms . In this study , we attempted to investigate whether vinpocetine protected against atherosclerotic development in apoE(-/-) mice and explore the underlying anti-atherogenic mechanisms in macrophages . METHODOLOGY/PRINCIPAL FINDINGS : Vinpocetine markedly decreased atherosclerotic lesion size in apoE(-/-) mice measured by oil red O . Masson 's trichrome staining and immunohistochemical analyses revealed that vinpocetine significantly increased the thickness of fibrous cap , reduced the size of lipid-rich necrotic core and attenuated inflammation . In vitro experiments exhibited a significant decrease in monocyte adhesion treated with vinpocetine . Further , active P01375 -α , P05231 , monocyte chemoattractant protein-1 and matrix metalloproteinase-9 expression induced by ox-LDL were attenuated by vinpocetine in a dose-dependent manner . Similarly , ox-LDL-induced reactive oxygen species were significantly repressed by vinpocetine . Both western blot and luciferase activity assay showed that vinpocetine inhibited the enhanced Akt , IKKα/β , IκBα phosphorylation and NF-κB activity induced by ox-LDL , and the inhibition of NF-κB activity was partly caused by Akt dephosphorylation . However , knockdown of Q01064 did not affect Akt , IKKα/β and IκBα phosphorylation . CONCLUSIONS : These results suggest that vinpocetine exerts anti-atherogenic effects through inhibition of monocyte adhesion , oxidative stress and inflammatory response , which are mediated by Akt/NF-κB dependent pathway but independent of PDE1 blockade in macrophages . P04035 inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells . OBJECTIVE : Q9UEF7 is thought to play a critical role in the development of age-related disorders including arteriosclerosis . Statins may exert vascular protective effects , independent of the lowering of plasma cholesterol levels . We investigated the impact of statins on mRNA expression of the age-suppressor gene , klotho in mIMCD3 cells . METHODS AND RESULTS : Q9UEF7 mRNA levels were evaluated with real-time RT-PCR . DB01076 and pitavastatin increased the expression of klotho mRNA in a dose-dependent manner . This stimulatory effect was abolished by the addition of mevalonate , GGPP and FPP , essential molecules for isoprenylation of the small GTPase Rho . As was the case with the statin treatment , inhibition of Rho-kinase by Y27632 up-regulated klotho mRNA . In contrast to the statin treatment , stimulation with angiotensin II down-regulated klotho mRNA expression without obvious morphological changes . Furthermore , pretreatment with atorvastatin blunted the angiotensin II-induced response and ameliorated the decrease in klotho mRNA expression towards basal levels . RhoA activity was further evaluated by detection of its translocation . Angiotensin II activated RhoA , whereas statins potently inactivated RhoA and blocked RhoA activation by angiotensin II . CONCLUSION : Statins inactivate the RhoA pathway , resulting in over-expression of klotho mRNA , which may contribute to the novel pleiotropic effects of statins towards vascular protection . Does computational biology help us to understand the molecular phylogenetics and evolution of cluster of differentiation ( CD ) proteins ? Cluster of differentiation ( CD ) is a group of proteins with highly immunological and medical importance , and some are established therapeutics . These membrane proteins are used to investigate of cell surface molecules of blood cells especially WBC . We selected a population of fifteen members with most medical importance , which includes P06729 , P01730 , P06127 , P30203 , P09564 , P21926 , P08571 , CD16 , P15391 , P20273 , P10747 , P20138 , P16671 , P28907 , and P16070 and performed in silico analysis using algorithm analysis and mathematical models . The results suggest that LEU ( L ) is well aligned . CD16 is rooted with P20273 and likewise , P01730 is closely related to P16070 . Notably , highest number of highly conserved amino acids is recorded in P20273 . WebLogo were formed up to 350 amino acid position and DB00134 ( M ) is found to be tallest logo . Our results would be useful for upcoming researchers to obtain fundamental idea about the particular regions CD proteins which is having the structural and functional significance related to the evolutionary biology . HIV entry inhibitors : mechanisms of action and resistance pathways . Entry inhibitors represent a new generation of antivirals for the treatment of HIV infection . Several compounds which block the attachment of HIV gp120 to either the P01730 T cell receptor or the P51681 / P61073 co-receptors are currently in clinical development . Most of these compounds have different molecular structures and specific mechanisms of action . These agents are eagerly awaited by a growing number of patients carrying viruses resistant viruses to many of the current available reverse transcriptase and protease inhibitors . For enfuvirtide , the first and , so far , only entry inhibitor approved for clinical use , the main mechanism of resistance is the selection of changes within a 10 amino acid segment encompassing residues 36-45 within the Q96GN5 region of gp41 . For other entry inhibitors , multiple changes in different gp120 domains ( V1 , V2 , V3 , P06681 and C4 ) have been associated with loss of susceptibility to these agents , although in most cases with limited cross-resistance . Telmisartan , its potential therapeutic implications in cardiometabolic disorders . There is a growing body of evidence that the renin-angiotensin system ( DB01367 ) plays a pivotal role in the pathogenesis of cardiovascular diseases . Indeed , large clinical trials have demonstrated substantial benefit of the blockade of this system for cardiovascular-organ protection . 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 . Telmisartan will be a promising cardiometabolic sartan due to its unique peroxisome proliferator-activated receptor-gamma ( P37231 ) -inducing properties as well . In this review , we focused on telmisartan , and discussed its potential therapeutic implications in cardiometabolic disorders . 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 . 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 P04035 inhibitor , atorvastatin , promotes a Th2 bias and reverses paralysis in central nervous system autoimmune disease . Statins , 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors , which are approved for cholesterol reduction , may also be beneficial in the treatment of inflammatory diseases . DB01076 ( Lipitor ) was tested in chronic and relapsing experimental autoimmune encephalomyelitis , a P01730 (+) Th1-mediated central nervous system ( CNS ) demyelinating disease model of multiple sclerosis . Here we show that oral atorvastatin prevented or reversed chronic and relapsing paralysis . DB01076 induced P42226 phosphorylation and secretion of Th2 cytokines ( interleukin ( IL ) -4 , P05113 and P22301 ) and transforming growth factor ( TGF ) -beta . Conversely , Q14765 phosphorylation was inhibited and secretion of Th1 cytokines ( P60568 , IL-12 , interferon ( IFN ) -gamma and tumour necrosis factor ( P01375 ) -alpha ) was suppressed . DB01076 promoted differentiation of Th0 cells into Th2 cells . In adoptive transfer , these Th2 cells protected recipient mice from EAE induction . DB01076 reduced CNS infiltration and major histocompatibility complex ( MHC ) class II expression . Treatment of microglia inhibited P01579 -inducible transcription at multiple P33076 ( P33076 ) promoters and suppressed class II upregulation . DB01076 suppressed P01579 -inducible expression of P25942 , P33681 and P42081 co-stimulatory molecules . l-Mevalonate , the product of P04035 , reversed atorvastatin 's effects on antigen-presenting cells ( P25054 ) and T cells . DB01076 treatment of either P25054 or T cells suppressed antigen-specific T-cell activation . Thus , atorvastatin has pleiotropic immunomodulatory effects involving both P25054 and T-cell compartments . Statins may be beneficial for multiple sclerosis and other Th1-mediated autoimmune diseases . Molecular basis of the structural stability of a Top7-based scaffold at extreme pH and temperature conditions . The development of stable biomolecular scaffolds that can tolerate environmental extremes has considerable potential for industrial and defense-related applications . However , most natural proteins are not sufficiently stable to withstand non-physiological conditions . We have recently engineered the de novo designed Top7 protein to specifically recognize the glycoprotein P01730 by insertion of an eight-residue loop . The engineered variant exhibited remarkable stability under chemical and thermal denaturation conditions . In the present study , far-UV CD spectroscopy and explicit-solvent MD simulations are used to investigate the structural stability of Top7 and the engineered variant under extreme conditions of temperature and pH . Circular dichroism measurements suggest that the engineered variant Top7( P21554 ) , like Top7 , retains its structure at high temperatures . Changes in CD spectra suggest that there are minor structural rearrangements between neutral and acidic environments for both proteins but that these do not make the proteins less stable at high temperatures . The anti-parallel beta-sheet is well conserved within the timescale simulated whereas there is a decrease of helical content when low pH and high-temperature conditions are combined . Concerted alanine mutations along the alpha-helices of the engineered Top7 variant did not revert this trend when at pH 2 and 400K . The structural resilience of the anti-parallel beta-sheet suggests that the protein scaffold can accommodate varying sequences . The robustness of the Top7 scaffold under extreme conditions of pH and temperature and its amenability to production in inexpensive bacterial expression systems reveal great potential for novel biotechnological applications . 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 . The role of atorvastatin in regulating the immune response leading to vascular damage in a model of Kawasaki disease . Superantigens have been implicated in a number of diseases including Kawasaki disease ( KD ) , a multi-system vasculitis resulting in coronary artery aneurysms . We have characterized a murine disease model in which coronary arteritis is induced by a novel superantigen found in Lactobacillus casei cell wall extract ( LCWE ) . Using this animal model of KD , we have identified three pathogenic steps leading to coronary artery aneurysm formation . These steps include T cell activation and proliferation , production of the proinflammatory cytokine tumour necrosis factor ( P01375 ) -α and up-regulation of matrix metalloproteinase 9 ( P14780 ) , an elastolytic protease . In addition to their cholesterol-lowering effects , 3-hydroxy-3-methylglutaryl ( HMG ) coenzyme A ( DB01992 ) reductase inhibitors ( statins ) have pleotropic immunomodulatory properties . Thus , we examined the effect of atorvastatin in modulating each of these three critical pathogenic processes leading to aneurysm formation in the disease model . DB01076 inhibited lymphocyte proliferation in response to superantigen stimulation in a dose-dependent manner . This inhibition was also observed for production of soluble mediators of inflammation including interleukin ( IL ) -2 and P01375 -α . The inhibitory effect on proliferation was rescued completely by mevalonic acid , confirming that the mechanism responsible for this inhibitory activity on immune activation was inhibition of P04035 . Similarly , P01375 -α-induced P14780 production was reduced in a dose-dependent manner in response to atorvastatin . Inhibition of extracellular-regulated kinase ( P29323 ) phosphorylation appears to be the mechanism responsible for inhibition of P14780 production . In conclusion , atorvastatin is able to inhibit critical steps known to be important in the development of coronary aneurysms , suggesting that statins may have therapeutic benefit in patients with KD . Comparison of rimonabant and sibutramine treatment effects on food compulsion in rats . PURPOSE : To compare the therapeutic effect of rimonabant , a new drug which is a selective antagonist of P21554 receptors , with the sibutramine . METHODS : It is an experimental clinical trial , prospective , placebo controlled . Our test was performed in 38 rats , adults females with a hyper caloric diet . We collected their blood 3 times and weighted them once a week . We divided the rats in 3 groups : DB06155 , DB01105 and Control . Statistic analysis has been made through Q9UNW9 test , Tukey test and t Student test . RESULTS : The DB06155 group demonstrated a significant reduction of the weight increase in rats . The DB01105 group showed a significant reduction on blood glycemia compared to DB06155 group and Control group . CONCLUSIONS : DB06155 showed to be more effective than DB01105 by decreasing weight gain . DB01105 has been more effective than DB06155 and Control groups by decreasing the blood glycemia . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Effect of atorvastatin on endothelial function and inflammation in long-duration type 1 diabetic patients without coronary heart disease and arterial hypertension . AIM : We evaluated the ability of atorvastatin , an P04035 inhibitor , to affect endothelial function and inflammation in long-duration ( > 10 years ) type 1 diabetes mellitus ( T1DM ) patients without coronary heart disease ( Q8NE62 ) and arterial hypertension ( AH ) . METHODS AND RESULTS : We randomized 204 Caucasians with long-duration T1DM into either the atorvastatin 40 mg/day plus hypolipaemic diet group ( n = 154 ) or the placebo plus hypolipaemic diet group ( n = 50 ) for 6 months . Endothelium-dependent flow-mediated ( FMD ) and endothelium-independent flow-mediated vasodilatation , serum levels of plasminogen activator inhibitor-1 ( P05121 ) , P04275 ( P04275 ) and high sensitivity P02741 ( hs-CRP ) were estimated before and after treatment . After 6 months of therapy , FMD was increased by 44 % in the atorvastatin plus diet group compared with the placebo plus diet group . Treatment with atorvastatin led to a significant reduction in levels of P05121 and hs-CRP ; however , the elevation of P04275 level was observed . In the placebo plus diet group , we observed a significant reduction in levels of hs-CRP but not of P04275 and P05121 . CONCLUSIONS : DB01076 improves endothelial function and reduces some proinflammatory and prothrombotic markers of atherosclerosis in T1DM patients without Q8NE62 and AH . The surprising effect of atorvastatin on serum P04275 levels in T1DM requires further study . DB02533 downregulates expression of cytokine-induced Fas and inducible nitric oxide synthase but not cytokine-enhanced surface antigens of rat islet cells . Autoimmune beta-cell destruction occurs directly by cell-mediated cytotoxicity or indirectly by cytokines released from infiltrating lymphocytes . Cytokines ( IL-1beta/ P01579 ) modify or induce expression of MHC antigens and P05362 on beta-cells which can lead to an improved binding of T-lymphocytes to beta-cells and finally to an enhanced cell-mediated cytotoxicity . Cytokines also induce Fas-expression and inducible nitric oxide synthase ( P35228 ) causing generation of nitric oxide ( NO ) which is toxic for beta-cells . The P35228 inhibitor aminoguanidine ( AG ) delays diabetes onset , but does not reduce diabetes incidence . We wanted to know whether AG inhibits cytokine-induced expression of Fas , MHC antigens and P05362 on beta-cells of LEW.1W and BB/OK rat islets after culture with IL-1beta/ P01579 . NO was completely inhibited by 5.0 mmol/L AG while 0.5 mmol/L had no inhibitory effect . AG downregulated Fas-expression on the surface of beta-cells . Cytokine-induced/enhanced expression of MHC class-II and P05362 was not affected by any AG concentration . AG syngergistically increased cytokine-induced enhancement of MHC class-I antigen density . AG possibly blocks the indirect pathway of beta-cell damage in vivo due to inhibition of Fas and P35228 and improves direct cell-mediated cytotoxicity due to drastic increased MHC class-I expression . Inhibition of only one pathway of beta-cell destruction is not sufficient to prevent diabetes . 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 . 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 . Mortality prediction using modern peptide biomarkers in hemodialysis patients -- a comparative analysis . BACKGROUND/AIMS : Determination of peptide biomarkers such as troponins , natriuretic peptides or the recently reported Q9GZV9 can be useful to identify hemodialysis patients with a high risk of mortality . However , it is desirable to focus on few robust parameters to warrant their routine application . METHODS : In a prospective cohort study with 239 prevalent hemodialysis patients we studied the prognostic significance of 10 simultaneously determined modern peptide biomarkers ( high sensitive troponin I and T , NT-pro- DB04899 , DB04899 , MR-pro- P01160 , MR-pro- P35318 , CT-pro-ET1 , copeptin , Q9GZV9 and a- Q9UEF7 ) and compared them with parameters traditionally associated with mortality ( PTH , Ca , Pi , albumin , CRP , cholesterol , AP ) . RESULTS : After a follow-up of 4 years , plasma concentration of troponins , natriuretic peptides , MR-pro- P35318 , Q9GZV9 as well as PTH , CRP , AP were significantly higher in deceased patients ( n=95 ) . Hazard ratios from cox regression on a continuous scale ( doubling of plasma concentration ) or relative in tertiles were highest for high sensitive troponins , followed by natriuretic peptides and MR-pro- P35318 ( 1.6-2.0 and 2.3-5.5 , resp. ) . C-indices were also highest for troponins ( 0.708-0.746 ) , followed by natriuretic peptides ( 0.706-0.731 ) . Traditional parameters had low c-indices ( 0.598-0.655 ) . Stepwise cox regression revealed that among all parameters troponin I , NT-pro- DB04899 , PTH and CRP remained independent predictors of mortality and a composite score had the highest c-index ( 0.799 [ 0.740-0.849 ] ) . CONCLUSIONS : Among peptide biomarkers high sensitive troponins and to a lesser extent natriuretic peptides are strong predictors of mortality in asymptomatic hemodialysis patients , followed by markers of mineral-bone disease and inflammation . Circulating P23582 ( P09543 ) rescues chondrodysplastic P09543 knockout mice from their impaired skeletal growth and early death . P23582 ( P09543 ) is a potent stimulator of endochondral bone growth through a subtype of membranous guanylyl cyclase receptor , P20594 . Although its two cognate natriuretic peptides , P01160 and DB04899 , are cardiac hormones produced from heart , P09543 is thought to act as an autocrine/paracrine regulator . To elucidate whether systemic administration of P09543 would be a novel medical treatment for chondrodysplasias , for which no drug therapy has yet been developed , we investigated the effect of circulating P09543 by using the P09543 transgenic mice with an increased circulating P09543 under the control of human serum amyloid P component promoter ( SAP-Nppc-Tg mice ) . SAP-Nppc-Tg mice developed prominent overgrowth of bones formed through endochondral ossification . In organ culture experiments , the growth of tibial explants of SAP-Nppc-Tg mice was not changed from that of their wild-type littermates , exhibiting that the stimulatory effect on endochondral bone growth observed in SAP-Nppc-Tg mice is humoral . Then we crossed chondrodysplastic P09543 -depleted mice with SAP-Nppc-Tg mice . Impaired endochondral bone growth in P09543 knockout mice were considerably and significantly recovered by increased circulating P09543 , followed by the improvement in not only their longitudinal growth but also their body weight . In addition , the mortality of P09543 knockout mice was greatly decreased by circulating P09543 . Systemic administration of P09543 might have therapeutic potential against not only impaired skeletal growth but also other aspects of impaired growth including impaired body weight gain in patients suffering from chondrodysplasias and might resultantly protect them from their early death .

MH_train_1402

MH_train_1402

MH_train_1402

interacts_with DB00783?

multiple_choice

DB04216 arrests G2/M phase and induces caspase-dependent cell death in U937 cells . DB04216 , a natural product derived from grapes , has been shown to prevent carcinogenesis in murine models . We report here that quercetin induces anti-proliferation and arrests G2/M phase in U937 cells . The G2/M phase accumulation was accompanied by an increase in the level of the cyclin B . In contrast , the level of the cyclin D , cyclin E , Q01094 , and Q14209 was marked decreased in quercetin-treated U937 cells . Removal of quercetin from the culture medium stimulates U937 cells to synchronously re-enter the cell cycle , decrease expression level of cyclin B , and increased the expression level of cyclin D and cyclin E . These data demonstrate that quercetin causes reversible G2/M phase arrest , which was related with dramatic changes in the level of cyclin B , cyclin D , and cyclin E. DB04216 -induced down-regulation of cyclin D and cyclin E was associated with suppression of transcriptional levels but not protein stability . In addition , quercetin-treated U937 cells showed DNA fragmentation , increased sub- P55008 population , and generated a 60kDa cleavage product of P98160 -gamma1 in a dose-dependent manner , which were significantly inhibited by z-VAD-fmk . These data clearly indicate that quercetin-induced apoptosis is associated with caspase activation . In summary , the growth inhibition of the quercetin is highly related to cell cycle arrest at the G2/M phase and induction of caspase-dependent apoptosis in human promonocytic U937 cells . Leptin promotes neointima formation and smooth muscle cell proliferation via NADPH oxidase activation and signalling in caveolin-rich microdomains . AIMS : P02649 ( apoE ) may act as a vasculoprotective factor by promoting plasma lipid clearance and cholesterol efflux . Moreover , apoE accumulates at sites of vascular injury and modulates the effect of growth factors on smooth muscle cells ( SMCs ) . Experimental data suggested that hypothalamic apoE expression is reduced in obesity and associated with leptin resistance . In this study , we examined the role of apoE in mediating the effects of leptin on vascular lesion formation . METHODS AND RESULTS : Leptin was administered to apoE knockout ( apoE-/- ) mice via osmotic pumps to increase its circulating levels . Morphometric analysis revealed that leptin did not alter neointima formation and failed to increase α-actin- or P12004 -immunopositive SMCs after vascular injury . Similar findings were obtained after analysis of atherosclerotic lesions . Comparison of apoE-/- , wild-type , or P01130 -/- mice and functional analyses in aortic SMCs from WT or apoE-/- mice or human arterial SMCs after treatment with small interfering (si)RNA or heparinase revealed that leptin requires the presence of apoE , expressed , secreted and bound to the cell surface , to fully activate leptin receptor signalling and to promote SMC proliferation and neointima formation . Mechanistically , leptin induced the phosphorylation and membrane translocation of caveolin (cav)-1 , and apoE down-regulation or caveolae disruption inhibited the leptin-induced p47phox activation , ROS formation and SMC proliferation . Finally , leptin failed to increase neointima formation in mice lacking cav-1 . CONCLUSION : Our findings suggest that apoE mediates the effects of leptin on vascular lesion formation by stabilizing cav-1-enriched cell membrane microdomains in SMCs , thus allowing NADPH oxidase assembly and ROS-mediated mitogenic signalling . Identification of DNA polymorphisms associated with the V type alpha1-antitrypsin gene . alpha1-Antitrypsin ( alpha1-AT ) is a highly polymorphic protein . The V allele of alpha1-AT has been shown to be associated with focal glomerulosclerosis ( FGS ) in Negroid and mixed race South African patients . To identify mutations and polymorphisms in the gene for the V allele of alpha1-AT in five South African patients with FGS nephrotic syndrome DNA sequence analysis and restriction fragment length polymorphisms of the coding exons were carried out . Four of the patients were heterozygous for the BstEII RFLP in exon III [ M1(Val213)(Ala213) ] and one patient was a M1(Ala213) homozygote . The mutation for the V allele was identified in exon II as DB00145 -148 ( GGG ) --> DB00125 ( AGG ) and in all patients was associated with a silent mutation at position 158 ( AAC --> P01009 ) . The patient who was homozygous for ( Ala213 ) also had a silent mutation at position 256 in exon III ( Q6IB77 --> GAC ) which was not present in any of the other four patients . Although the V allele of alpha1-AT is not associated with severe plasma deficiency , it may be in linkage disequilibrium with other genes on chromosome 14 that predispose to FGS . Furthermore , the associated silent mutation at position 158 and the Ala213 polymorphism are of interest , as these could represent an evolutionary intermediate between the M1(Ala213) and M1(Val213) subtypes . Evaluation of human astrocytoma and glioblastoma cell lines for nerve growth factor release . Nerve growth factor ( P01138 ) prevents degeneration of cholinergic neurons in the central nervous system ( CNS ) , and has potential as a therapeutic treatment for Alzheimer 's disease . The inability of P01138 to cross the blood-brain barrier has prompted pharmacological approaches investigating peripherally administered compounds that stimulate release of endogenous P01138 . This study describes the P01138 -releasing properties of six human astrocytoma and glioblastoma cell lines ( SW 1088 , SW 1783 and CRL 1718 astrocytomas , and U-138 , U-373 , and T98G glioblastomas ) . Using a highly specific two-site ELISA for human P01138 , basal P01138 release could be detected in all cell lines , with the lowest level in the T98G line ( approximately 80 pg P01138 /ml ) . Cell lines tested with a variety of compounds for 24 h in serum-free media demonstrated stimulation of P01138 release by distinct mechanisms . P01138 levels were markedly elevated ( up to 8-fold above vehicle-treated cells ) when stimulated with the cytokine interleukin-1 beta ( P01584 ) . Phorbol ester stimulated P01138 release 4-fold . DB01407 , 4-methyl catechol , and propentofylline had little activity , while 6-(4-hydroxybutyl)-2,3,5,-trimethyl-1,4,benzoquinone ( DB01157 ) , dexamethasone and 1,25-dihydroxyvitamin D3 elevated P01138 levels 3-fold . These data indicate differences in the ability of human astrocytoma and glioblastoma cells to release P01138 when stimulated with mechanistically distinct compounds . Loss of the candidate tumor suppressor Q14201 triggers acute cellular senescence via the P29323 - O15054 -p16(INK4a) signaling axis . The B-cell translocation gene 3 ( Q14201 ) is a member of the antiproliferative BTG gene family and a downstream target of p53 . Q14201 also binds and inhibits Q01094 . Although it connects functionally two major growth-regulatory pathways , the physiological role of Q14201 remains largely uncharacterized . Here , we present evidence that loss of Q14201 in normal cells induced cellular senescence , which was correlated with enhanced P29323 - P05412 signaling and elevated expression of the histone H3K27me3 demethylase O15054 / O15054 , leading to acute induction of p16(INK4a) . Importantly , we also found that Q14201 expression is specifically downregulated in prostate cancer , thus providing a physiological link with human cancers . Our data suggest that Q14201 may have a fail-safe role against tumorigenic progression . 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 . 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 . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . Systems biology of autosomal dominant polycystic kidney disease ( ADPKD ) : computational identification of gene expression pathways and integrated regulatory networks . To elucidate the molecular pathways that modulate renal cyst growth in ADPKD , we performed global gene profiling on cysts of different size ( < 1 ml , n = 5 ; 10-20 ml , n = 5 ; > 50 ml , n = 3 ) and minimally cystic tissue ( Q8IVS2 , n = 5 ) from five PKD1 human polycystic kidneys using Affymetrix HG-U133 Plus 2.0 arrays . We used gene set enrichment analysis to identify overrepresented signaling pathways and key transcription factors ( TFs ) between cysts and Q8IVS2 . We found down-regulation of kidney epithelial restricted genes ( e.g. nephron segment-specific markers and cilia-associated cystic genes such as P35680 , P08F94 , Q13099 and Q717R9 ) in the renal cysts . On the other hand , PKD1 cysts displayed a rich profile of gene sets associated with renal development , mitogen-mediated proliferation , cell cycle progression , epithelial-mesenchymal transition , hypoxia , aging and immune/inflammatory responses . Notably , our data suggest that up-regulation of Wnt/beta-catenin , pleiotropic growth factor/receptor tyrosine kinase ( e.g. IGF/ P08069 , FGF/FGFR , P01133 / P00533 , P15692 /VEGFR ) , G-protein-coupled receptor ( e.g. PTGER2 ) signaling was associated with renal cystic growth . By integrating these pathways with a number of dysregulated networks of TFs ( e.g. P11831 , MYC , Q01094 , P16220 , Q9UJU2 , P36402 , P35680 / P20823 and P41235 ) , our data suggest that epithelial dedifferentiation accompanied by aberrant activation and cross-talk of specific signaling pathways may be required for PKD1 cyst growth and disease progression . Pharmacological modulation of some of these signaling pathways may provide a potential therapeutic strategy for ADPKD . Tumour suppressor p53 and Rb genes in human hepatocellular carcinoma . Aberrations of the p53 and Rb tumour suppressor genes were examined in 12 human hepatocellular carcinoma ( HCC ) -derived cell lines from different geographic areas and 9 local HCCs by restriction fragment length polymorphisms ( RFLP ) , polymerase chain reaction-single-strand conformation polymorphisms ( PCR-SSCP ) and DNA sequencing . The relationships between genetic changes and hepatitis B virus ( HBV ) DNA integration in samples were compared . None of the cell lines and tumours showed structural changes in the Rb gene , while 6 cell lines and 2 tumours had mutation or deletion in exons 5 to 8 of p53 . Mutations include an AGG --> AGT ( DB00125 --> DB00133 ) transversion at codon 249 in P98160 /PRF/5 and Mahlavu , an P01009 --> AAA ( DB00174 --> DB00151 ) transversion at codon 200 in TONG/HCC , an P29372 --> GAG ( Lys --> DB00142 ) transition at codon 139 in HCC-T , a CAT --> Q16880 ( DB00117 --> DB00125 ) transition at codon 214 in SC4 , and a CCC --> CTC ( Pro --> DB00149 ) transition at codon 250 in SC8 . In Huh4 , an 18-bp deletion from codon 264 to 270 resulted in loss of DB00149 - DB00145 - DB00125 - DB00174 - DB00133 - DB00120 from the amino acid sequences 265 to 270 , whereas Hep3B had a 7-kb deletion after exon 7 of p53 . Our data indicate that whereas Rb may not have pleiotropic effects on HCC , p53 aberrations are frequently involved in hepatocarcinogenesis . Further , HBV infection appears to be unrelated to the micro-genetic changes of p53 . The G to T codon-249-mutation is consistent with HCCs arising from areas at high risk for both aflatoxin B1 ( AFB1 ) exposure and HBV infection . 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 . Growth factors and melanomas . Understanding the growth constraints imposed on normal human melanocytes may help to elucidate the processes conferring growth advantage to melanoma cells . Several synergistic growth factors have been identified for normal human melanocytes . They include fibroblast growth factors ( FGF ) , hepatocyte growth factor/scatter factor , mast/stem cell growth factor , and the neuropeptides endothelin-1 , 2 and 3 ( ET-1 , P20800 , P14138 ) . From this group of peptides , only basic FGF ( P09038 / P09038 ) appears , so far , to play a role in autonomous growth of melanoma cells . Aberrant expression of P09038 is due to activation of an otherwise repressed gene by a mechanism that may involve the transcriptional activity of wild-type p53 . The growth factors and activated receptors aberrantly expressed in melanoma cells act in concert with molecules that control cell cycle progression . These proteins bind to , and regulate cyclin-dependent kinase ( CDK ) , such as P11802 , responsible for phosphorylation of retinoblastoma ( RB ) and dissociation of RB- Q01094 inhibitory complexes , thereby allowing progression through the cell cycle . Constitutive P11802 activity in melanomas may be the results of inactivation of the negative regulators known as CDK inhibitor p16INK4 , and/or P38936 ; and/or overexpression of cyclin D , the positive P11802 regulator . This complex set of changes in melanoma cells can lift growth constraints by inducing unregulated expression of genes promoting transition from GI to S phase of the cell cycle . Suppression of P38398 sensitizes cells to proteasome inhibitors . P38398 is a multifunctional protein best known for its role in DNA repair and association with breast and ovarian cancers . To uncover novel biologically significant molecular functions of P38398 , we tested a panel of 198 approved and experimental drugs to inhibit growth of MDA-MB-231 breast cancer cells depleted for P38398 by siRNA . 26S proteasome inhibitors bortezomib and carfilzomib emerged as a new class of selective P38398 -targeting agents . The effect was confirmed in HeLa and U2OS cancer cell lines using two independent siRNAs , and in mouse embryonic stem ( ES ) cells with inducible deletion of Brca1 . DB00188 treatment did not cause any increase in nuclear foci containing phosphorylated histone P16104 , and knockdown of P51587 did not entail sensitivity to bortezomib , suggesting that the DNA repair function of P38398 may not be directly involved . We found that a toxic effect of bortezomib on P38398 -depleted cells is mostly due to deregulated cell cycle checkpoints mediated by P06400 -E2F pathway and Q12888 . Similar to P38398 , depletion of P06400 also conferred sensitivity to bortezomib , whereas suppression of Q01094 or Q12888 together with P38398 reduced induction of apoptosis after bortezomib treatment . A gene expression microarray study identified additional genes activated by bortezomib treatment only in the context of inactivation of P38398 including a critical involvement of the O75460 -mediated unfolded protein response . Our data indicate that P38398 has a novel molecular function affecting cell cycle checkpoints in a manner dependent on the 26S proteasome activity . A curated database of miRNA mediated feed-forward loops involving MYC as master regulator . BACKGROUND : The MYC transcription factors are known to be involved in the biology of many human cancer types . But little is known about the Myc/microRNAs cooperation in the regulation of genes at the transcriptional and post-transcriptional level . METHODOLOGY/PRINCIPAL FINDINGS : Employing independent databases with experimentally validated data , we identified several mixed microRNA/Transcription Factor Feed-Forward Loops regulated by Myc and characterized completely by experimentally supported regulatory interactions , in human . We then studied the statistical and functional properties of these circuits and discussed in more detail a few interesting examples involving Q01094 , P60484 , P06400 and P15692 . CONCLUSIONS/SIGNIFICANCE : We have assembled and characterized a catalogue of human mixed Transcription Factor/microRNA Feed-Forward Loops , having Myc as master regulator and completely defined by experimentally verified regulatory interactions . 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 . 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 . 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 . Use of RNA interference to elucidate the effect of P04198 on cell cycle in neuroblastoma . BACKGROUND : P04198 amplification marks poor prognosis in neuroblastoma ( NB ) tumors . In evaluating the mechanisms by which retinoic acid ( RA ) or nerve growth factor ( P01138 ) decrease cell number in P04198 amplified NB cells , we have identified a number of proteins whose expression either decreases ( E2F , P06493 , Q00534 , cyclin dependent kinase activity ) or increases ( p27 ) in association with a decrease in P04198 expression . However , it was still unclear which were P04198 dependent effects or not . PROCEDURE : This study aimed to determine which changes in cell cycle gene expression are modulated as a consequence of the decrease in P04198 . We silenced P04198 expression using siRNA targeted to the coding region of P04198 . Then , by using siRNA transient transfections , we analyzed the change of cell cycle related genes and cell cycle in P04198 amplified NB cell lines . RESULTS : We demonstrate that expression of P04198 can be suppressed by almost 60 % in P04198 amplified NB cell using siRNAs targeted to P04198 . Functionally , the decrease in P04198 leads to a decrease in cells in the S-phase of the cell cycle . Decreases in P04198 are associated with decreases in Q01094 -2 and Q02363 along with increases in p27 protein levels by post-transcriptional modification . Moreover , we find that a decrease in P04198 is accompanied by a decrease in cdk6 mRNA and protein expression . CONCLUSIONS : These results show that E2F and Q02363 expression is associated with P04198 regulation and that cdk6 is a possible new transcriptional target of P04198 . The BH3 mimetic DB05764 synergizes with the Q02750 /2 inhibitor selumetinib/AZD6244 to promote O43521 -dependent tumour cell death and inhibit acquired resistance . Tumour cells typically exhibit a G(1) cell cycle arrest in response to the Q02750 /2 [ mitogen-activated protein kinase/ P29323 ( extracellular-signal-regulated kinase ) kinase 1/2 ] inhibitor selumetinib , but do not die , and thus they acquire resistance . In the present study we examined the effect of combining selumetinib with the BH3 [ P10415 ( B-cell lymphoma 2 ) homology domain 3 ] -mimetic P10415 inhibitor DB05764 . Although either drug alone caused little tumour cell death , the two agents combined to cause substantial caspase-dependent cell death and inhibit long-term clonogenic survival of colorectal cancer and melanoma cell lines with P15056 (V600E) or DB01367 mutations . This cell death absolutely required Q07812 ( P10415 -associated X protein ) and was inhibited by RNAi ( RNA interference ) -mediated knockdown of O43521 ( P10415 -interacting mediator of cell death ) in the P15056 (V600E)-positive COLO205 cell line . When colorectal cancer cell lines were treated with selumetinib plus DB05764 we observed a striking reduction in the incidence of cells emerging with acquired resistance to selumetinib . Similar results were observed when we combined DB05764 with the P15056 (V600E)-selective inhibitor PLX4720 , but only in cells expressing P15056 (V600E) . Finally , cancer cells in which acquired resistance to selumetinib arises through P15056 (V600E) amplification remained sensitive to DB05764 , whereas selumetinib-resistant HCT116 cells ( P01116 (G13D) amplification ) were cross-resistant to DB05764 . Thus the combination of a P10415 inhibitor and an P27361 /2 pathway inhibitor is synthetic lethal in P27361 /2-addicted tumour cells , delays the onset of acquired resistance and in some cases overcomes acquired resistance to selumetinib . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours .

MH_train_1403

MH_train_1403

MH_train_1403

interacts_with DB00977?

multiple_choice

Dietary soy protein isolate attenuates metabolic syndrome in rats via effects on Q07869 , LXR , and SREBP signaling . To determine the effects of feeding soy or isoflavones on lipid homeostasis in early development , weanling rats were fed AIN-93G diets made with casein , soy protein isolate ( SPI+ ) , isoflavone-reduced SPI+ ( SPI- ) , or casein supplemented with genistein or daidzein for 14 d . PPARalpha-regulated genes and proteins involved in fatty acid degradation were upregulated by SPI+ ( P < 0.05 ) accompanied by increased promoter binding and expression of PPARalpha mRNA ( P < 0.05 ) . Feeding SPI- or pure isoflavones did not alter PPARalpha-regulated pathways . SPI+ feeding had similar effects on PPARgamma signaling . SPI+ , SPI- , and casein plus isoflavones all increased liver Q9UBH6 (LXR)alpha-regulated genes and enzymes involved in cholesterol homeostasis . Feeding SPI+ increased promoter binding of LXRalpha , expression of the transcription factor mRNA , and protein ( P < 0.05 ) . In a second experiment , male Sprague-Dawley rats were fed casein diets from postnatal d ( P01160 ) 24 to PND64 or were fed high-fat Western diets containing 5 g x kg(-1) cholesterol made with either casein or SPI+ . P01308 resistance , steatosis , and hypercholesterolemia in the Western diet-fed rats were partially prevented by SPI+ ( P < 0.05 ) . Nuclear sterol receptor element binding protein ( SREBP ) -1c protein and mRNA and protein expression of enzymes involved in fatty acid synthesis were increased by feeding Western diets containing casein but not SPI+ ( P < 0.05 ) . These data suggest that activation of Q07869 and LXR signaling and inhibition of SREBP-1c signaling may contribute to insulin sensitization and improved lipid homeostasis in SPI+-fed rats after consumption of diets high in fat and cholesterol . Nucleolar exit of O76064 and P38398 in response to DNA damage . The induction of DNA double-strand breaks ( DSBs ) elicits a plethora of responses that redirect many cellular functions to the vital task of repairing the injury , collectively known as the DNA damage response ( DDR ) . We have found that , in the absence of DNA damage , the DSB repair factors O76064 and P38398 are associated with the nucleolus . Shortly after exposure of cells to γ-radiation , O76064 and P38398 translocated from the nucleolus to damage foci , a traffic that was reverted several hours after the damage . O76064 interacted through its FHA domain with the ribosomal protein P08865 , and knockdown of P08865 caused a depletion of nucleolar O76064 and P38398 , suggesting that the interaction of O76064 with P08865 is critical for the nucleolar localization of these DDR factors . Knockdown of P08865 or O76064 impaired bulk protein translation , as did γ-irradiation , the latter being partially countered by overexpression of exogenous O76064 . Our results suggest that O76064 and P38398 are anchored to the nucleolus through reversible interactions with P08865 and that , in addition to its known functions in DDR , O76064 may play a role in protein synthesis , possibly linking the nucleolar exit of this factor to the attenuation of protein synthesis in response to DNA damage . 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 . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . [ Progress in molecularly targeted therapies for acute myeloid leukemia ] . Genetic abnormalities including specific point mutations have recently been confirmed by applying comprehensive genome sequencing analyses . Molecular targeting therapies , which focus on the mutated proteins and over-expressed proteins in acute myeloid leukemia ( AML ) cells , are now being developed in clinical studies and/or based on in vitro analyses . This manuscript summarizes the genetic abnormalities in AML cells and some of the current molecular targeting therapies including P36888 inhibitors ( e.g. DB05213 ; Quizartinib ) , Polo like kinase 1 ( P53350 ) inhibitors ( e.g. BI-6727 ; Volasertib ) , P48735 inhibitors ( e.g. AG-221 ) , and O14980 inhibitors ( e.g. KPT-330 ; Selinexor ) . T cells targeting a neuronal paraneoplastic antigen mediate tumor rejection and trigger CNS autoimmunity with humoral activation . Paraneoplastic neurologic diseases ( P01160 ) involving immune responses directed toward intracellular antigens are poorly understood . Here , we examine immunity to the P01160 antigen Nova2 , which is expressed exclusively in central nervous system ( CNS ) neurons . We hypothesized that ectopic expression of neuronal antigen in the periphery could incite P01160 . In our C57BL/6 mouse model , CNS antigen expression limits antigen-specific P01730 + and CD8+ T-cell expansion . Chimera experiments demonstrate that this tolerance is mediated by antigen expression in nonhematopoietic cells . CNS antigen expression does not limit tumor rejection by adoptively transferred transgenic T cells but does limit the generation of a memory population that can be expanded upon secondary challenge in vivo . Despite mediating cancer rejection , adoptively transferred transgenic T cells do not lead to paraneoplastic neuronal targeting . Preliminary experiments suggest an additional requirement for humoral activation to induce CNS autoimmunity . This work provides evidence that the requirements for cancer immunity and neuronal autoimmunity are uncoupled . Since humoral immunity was not required for tumor rejection , B-cell targeting therapy , such as rituximab , may be a rational treatment option for P01160 that does not hamper tumor immunity . 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 . Founder effect and estimation of the age of the French Gypsy mutation associated with Glanzmann thrombasthenia in Manouche families . The c.1544+1G > A substitution at the 5' splice donor site of intron 15 of the P08514 gene , called the French Gypsy mutation , causes Glanzmann thrombasthenia , an inherited hemorrhagic disorder transmitted as an autosomal recessive trait and characterized by an altered synthesis of the platelet αIIbβ3 integrin . So far , this mutation has only been found in affected individuals originating from French Manouche families , strongly suggesting a founder effect . Our goal was to investigate the origin of the French Gypsy mutation . We estimated the age of the mutation by a likelihood-based method that uses the length of the shared haplotypes among a set of patients . For this , we genotyped 23 individuals of Manouche origin ; consisting of 9 Glanzmann thrombasthenia patients homozygous for the French Gypsy mutation , 6 heterozygous carriers and 8 homozygous wild-type individuals . They were genotyped for four single-nucleotide polymorphisms using high-resolution melting curve analysis , and for two CA repeats in the P38398 and P10827 genes at chromosome 17 , using fragment analysis gels . We found that a haplotype of five polymorphic loci covering a 4-cM region was strongly associated with the French Gypsy mutation , suggesting a founder effect . The estimated age of this founder mutation was 300-400 years ( range 255-552 years ) . Thus , all carriers of the French Gypsy mutation c.1544+1G > A at intron 15 descended from a common ancestor 300-400 years ago . 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 . O96028 deregulation affects cell cycle progression and adhesion regulons in t(4;14) myeloma plasma cells . BACKGROUND : The recurrent immunoglobulin translocation , t(4;14)(p16;q32) occurs in 15 % of multiple myeloma patients and is associated with poor prognosis , through an unknown mechanism . The t(4;14) up-regulates fibroblast growth factor receptor 3 ( P22607 ) and multiple myeloma Q01105 domain ( O96028 ) genes . The involvement of O96028 in the pathogenesis of t(4;14) multiple myeloma and the mechanism or genes deregulated by O96028 upregulation are still unclear . DESIGN AND METHODS : The expression of O96028 was analyzed using a novel antibody . The involvement of O96028 in t(4;14) myelomagenesis was assessed by small interfering RNA mediated knockdown combined with several biological assays . In addition , the differential gene expression of O96028 -induced knockdown was analyzed with expression microarrays . O96028 gene targets in primary patient material was analyzed by expression microarrays . RESULTS : We found that O96028 isoforms are expressed in multiple myeloma cell lines , being exclusively up-regulated in t(4;14)-positive cells . Suppression of O96028 expression affected cell proliferation by both decreasing cell viability and cell cycle progression of cells with the t(4;14) translocation . These findings were associated with reduced expression of genes involved in the regulation of cell cycle progression ( e.g. P30279 , P51959 , P38398 , O14965 and O14757 ) , apoptosis ( P29466 , P49662 and O43524 ) and cell adhesion ( Q13443 and Q14126 ) . Furthermore , we identified genes involved in the latter processes that were differentially expressed in t(4;14) multiple myeloma patient samples . CONCLUSIONS : In conclusion , dysregulation of O96028 affects the expression of several genes involved in the regulation of cell cycle progression , cell adhesion and survival . A cardiac pathway of cyclic GMP-independent signaling of guanylyl cyclase A , the receptor for atrial natriuretic peptide . Cardiac atrial natriuretic peptide ( P01160 ) regulates arterial blood pressure , moderates cardiomyocyte growth , and stimulates angiogenesis and metabolism . P01160 binds to the transmembrane guanylyl cyclase ( GC ) receptor , P16066 , to exert its diverse functions . This process involves a cGMP-dependent signaling pathway preventing pathological [Ca(2+)](i) increases in myocytes . In chronic cardiac hypertrophy , however , P01160 levels are markedly increased and P16066 /cGMP responses to P01160 are blunted due to receptor desensitization . Here we show that , in this situation , P01160 binding to P16066 stimulates a unique cGMP-independent signaling pathway in cardiac myocytes , resulting in pathologically elevated intracellular Ca(2+) levels . This pathway involves the activation of Ca(2+)-permeable transient receptor potential canonical 3/6 ( Q13507 / P13671 ) cation channels by P16066 , which forms a stable complex with Q13507 / P13671 channels . Our results indicate that the resulting cation influx activates voltage-dependent L-type Ca(2+) channels and ultimately increases myocyte Ca(2)(+)(i) levels . These observations reveal a dual role of the P01160 / P16066 -signaling pathway in the regulation of cardiac myocyte Ca(2+)(i) homeostasis . Under physiological conditions , activation of a cGMP-dependent pathway moderates the Ca(2+)(i)-enhancing action of hypertrophic factors such as angiotensin II . By contrast , a cGMP-independent pathway predominates under pathophysiological conditions when P16066 is desensitized by high P01160 levels . The concomitant rise in [Ca(2+)](i) might increase the propensity to cardiac hypertrophy and arrhythmias . The O14980 nuclear export protein in normal development and disease . O14980 ( Chromosomal Maintenance 1 , also known as Exportin 1 ) is the major mammalian export protein that facilitates the transport of large macromolecules including RNA and protein across the nuclear membrane to the cytoplasm . The gene encoding O14980 was originally identified in yeast as required to maintain higher order chromosome structure . In mammalian cells , O14980 was found to bind several nuclear pore proteins hence its role in nuclear-cytosolic transport was discovered . In addition to nuclear-cytosolic transport , O14980 also plays a role in centrosome duplication and spindle assembly , especially in response to DNA damage . The crystal structure of O14980 suggests a complex protein that binds the Ran protein bound to GTP , allowing for a conformational change that facilitates binding to different cargo proteins through a nuclear export signal ( P48681 ) . Included in the cadre of cargo are multiple tumor suppressor and oncoproteins as p53 , P38398 , Survivin , P06748 , and P25054 , which function in the nucleus to regulate transcription or aid in chromosomal assembly and movement . An imbalance in the cytosolic level of these proteins has been observed in cancer cells , resulting in either inactivation ( tumor suppressor ) or an excess of anti-apoptotic activity ( oncoprotein ) . Thus , the concept of inhibiting O14980 has been explored as a potential therapeutic intervention . Indeed , inhibition of O14980 by a variety of small molecules that interfere with cargo- P48681 binding results in cancer cell death . Whether all of these proteins together are responsible for this phenotype or whether specific proteins are required for this effect is unclear at this time . 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 . 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 . Thermal stability of the human immunodeficiency virus type 1 ( HIV-1 ) receptors , P01730 and P61073 , reconstituted in proteoliposomes . BACKGROUND : The entry of human immunodeficiency virus ( HIV-1 ) into host cells involves the interaction of the viral exterior envelope glycoprotein , gp120 , and receptors on the target cell . The HIV-1 receptors are P01730 and one of two chemokine receptors , P51681 or P61073 . METHODOLOGY/PRINCIPAL FINDINGS : We created proteoliposomes that contain P01730 , the primary HIV-1 receptor , and one of the coreceptors , P61073 . Antibodies against P01730 and P61073 specifically bound the proteoliposomes . P48061 , the natural ligand for P61073 , and the small-molecule P61073 antagonist , DB06809 , bound the proteoliposomes with affinities close to those associated with the binding of these molecules to cells expressing P61073 and P01730 . The HIV-1 gp120 exterior envelope glycoprotein bound tightly to proteoliposomes expressing only P01730 and , in the presence of soluble P01730 , bound weakly to proteoliposomes expressing only P61073 . The thermal stability of P01730 and P61073 inserted into liposomes was examined . Thermal denaturation of P61073 followed second-order kinetics , with an activation energy ( E(a) ) of 269 kJ/mol ( 64.3 kcal/mol ) and an inactivation temperature ( T(i) ) of 56°C . Thermal inactivation of P01730 exhibited a reaction order of 1.3 , an E(a) of 278 kJ/mol ( 66.5 kcal/mol ) , and a T(i) of 52.2°C . The second-order denaturation kinetics of P61073 is unusual among G protein-coupled receptors , and may result from dimeric interactions between P61073 molecules . CONCLUSIONS/SIGNIFICANCE : Our studies with proteoliposomes containing the native HIV-1 receptors allowed an examination of the binding of biologically important ligands and revealed the higher-order denaturation kinetics of these receptors . P01730 / P61073 -proteoliposomes may be useful for the study of virus-target cell interactions and for the identification of inhibitors . Arterial reocclusion and persistent distal occlusion after thrombus aspiration . BACKGROUND AND PURPOSE : Early reocclusion of intracranial arteries can lead to poor clinical outcome . We report reocclusion detection after endovascular clot aspiration , followed by administration of P08514 -IIIa antagonist under continuous ultrasound monitoring . CASE DESCRIPTION : A 73-year-old man developed the right middle cerebral artery ( MCA ) occlusion with NIHSS 17 points , 6 days after aortic valve replacement . Recanalization was achieved with Penumbra™ system and reocclusion was detected with transcranial Doppler ( P24386 ) 30 minutes postcompletion of intra-arterial procedure . Proximal recanalization was achieved with the second thrombus aspiration while M2 MCA occlusion persisted beyond the reach of the device . Intravenous abciximab was administered under continuous P24386 monitoring . Recanalization with Thrombolysis in Brain Ischemia ( TIBI ) flow grade 4 was observed at 60 minutes postintervention accompanied with clinical recovery to NIHSS 3 points . DB00054 was given for 12 hours with no hemorrhagic transformation on repeat CT scan . Patient was discharged home with mild left pronator drift and facial droop , and his modified ranking score was 1 at 6-week follow-up visit . CONCLUSIONS : Early arterial reocclusion can occur after successful thrombus aspiration while P08514 -IIIa antagonist administration may lead to subsequent recanalization of persisting distal occlusions not amenable to mechanical removal . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . 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 . 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 . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment .

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MH_train_1404

interacts_with DB00215?

multiple_choice

The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . Alloreactive memory T cells are responsible for the persistence of graft-versus-host disease . Graft-vs-host disease ( GVHD ) is caused by a donor T cell anti-host reaction that evolves over several weeks to months , suggesting a requirement for persistent alloreactive T cells . Using the C3H.SW anti-C57BL/6 ( B6 ) mouse model of human GVHD directed against minor histocompatibility Ags , we found that donor CD8(+) T cells secreting high levels of P01579 in GVHD B6 mice receiving C3H.SW naive CD8(+) T cells peaked by day 14 , declined by day 28 after transplantation , and persisted thereafter , corresponding to the kinetics of a memory T cell response . Donor CD8(+) T cells recovered on day 42 after allogeneic bone marrow transplantation expressed the phenotype of P16070 (high)CD122(high)CD25(low) , were able to homeostatically survive in response to P60568 , P13232 , and P40933 and rapidly proliferated upon restimulation with host dendritic cells . Both allogeneic effector memory ( P16070 (high)CD62L(low) ) and central memory ( P16070 (high)CD62L(high) ) CD8(+) T cells were identified in B6 mice with ongoing GVHD , with effector memory CD8(+) T cells as the dominant ( > 80 % ) population . Administration of these allogeneic memory CD8(+) T cells into secondary B6 recipients caused virulent GVHD . A similar allogeneic memory P01730 (+) T cell population with the ability to mediate persistent GVHD was also identified in BALB/b mice receiving minor histocompatibility Ag-mismatched B6 T cell-replete bone marrow transplantation . These results indicate that allogeneic memory T cells are generated in vivo during GVH reactions and are able to cause GVHD , resulting in persistent host tissue injury . Thus , in vivo blockade of both alloreactive effector and memory T cell-mediated host tissue injury may prove to be valuable for GVHD prevention and treatment . Q13224 subunit selective DB01221 antagonists inhibit neurotoxic effect of alcohol-withdrawal in primary cultures of rat cortical neurones . N-Methyl-D-aspartate ( DB01221 ) receptor-mediated glutamatergic neurotransmission is thought to play a central role in the development of alcohol dependence and this alteration is supposed to be due to a differential up-regulation of the Q13224 type of subunits . In this work , we examined the effect of some known ( CP-101,606 ; CI-1041 and Co-101,244 ) and novel indole-2-carboxamide derivative Q13224 subunit selective DB01221 receptor antagonists ( SSNAs ) ( RG-13579 and RG-1103 ) on the neurotoxic effect of withdrawal in ethanol pre-treated cultures of rat cortical neurones . The extent of neurotoxicity was estimated by measuring the activity of lactate dehydrogenase ( LDH ) that was released into the culture medium during the 24h withdrawal period . Here , we demonstrate that Q13224 SSNAs given in the course of the withdrawal potently reduced the LDH release in ethanol pre-treated cultures . One of our novel compound , RG-1103 , proved to be more potent than the reference Q13224 SSNAs tested in this work having similar potency as the most potent but non-subunit selective DB01221 receptor antagonist dizocilpine ( MK-801 ) . DB00659 , a currently used therapeutic drug for the treatment of alcoholism was also effective although only in high micromolar concentrations . According to these observations , Q13224 SSNAs are potent inhibitors of ethanol-withdrawal-induced neurotoxicity and considering that these agents have acceptable side effect profiles , they could be promising therapeutic candidates in the pharmacotherapy for physical signs of acute alcohol-withdrawal and associated neuronal damage . Regulation of apoptosis by tyrosine-containing domains of IL-4R alpha : Y497 and Y713 , but not the P42226 -docking tyrosines , signal protection from apoptosis . P05112 is a cytokine with important antiapoptotic activity . We have analyzed the role that tyrosine-containing domains within the cytoplasmic tail of IL-4R alpha play in P05112 -mediated protection from apoptosis . 32D cells expressing a wt huIL-4R alpha or one truncated at aa 557 were protected by huIL-4 from apoptosis while cells expressing a receptor truncated at aa 657 were not , suggesting that the carboxyl-terminal domain signals protection from apoptosis . However , changing Y713 within this region to phenylalanine had no effect . To analyze the contribution of tyrosine-containing domains independently , we transplanted regions of the huIL-4R alpha to a truncated form of the huIL-2R beta that could not signal protection from apoptosis . Transplantation of the huIL-4R alpha domains containing Y497 or Y713 partially prevented cell death and together signaled protection from apoptosis in response to P60568 as well as the wt IL-2R beta . Mutation of Y497 and Y713 to phenylalanine inhibited protection . In contrast , transplantation of the domain containing the potential P42226 -docking tyrosines alone had no effect , yet it inhibited the protection mediated by the other domains . Although IL-4R alpha signals Shc and SH2-containing inositol phosphatase ( Q92835 ) phosphorylation , we could not establish an association between their activation and protection from apoptosis . Taken together , this study suggests that the domains of the huIL-4R alpha containing Y497 and Y713 positively regulate protection from apoptosis while the domain containing the P42226 docking sites suppresses this protection , and that additional signaling molecules other than insulin receptor substrate-1 ( P35568 ) , Shc , or Q92835 may be involved in antiapoptotic signaling . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Aerosol vaccination with AERAS-402 elicits robust cellular immune responses in the lungs of rhesus macaques but fails to protect against high-dose Mycobacterium tuberculosis challenge . Development of a vaccine against pulmonary tuberculosis may require immunization strategies that induce a high frequency of Ag-specific P01730 and CD8 T cells in the lung . The nonhuman primate model is essential for testing such approaches because it has predictive value for how vaccines elicit responses in humans . In this study , we used an aerosol vaccination strategy to administer AERAS-402 , a replication-defective recombinant adenovirus ( rAd ) type 35 expressing Mycobacterium tuberculosis Ags Ag85A , Ag85B , and TB10.4 , in bacillus Calmette-Guérin ( BCG ) -primed or unprimed rhesus macaques . Immunization with BCG generated low purified protein derivative-specific P01730 T cell responses in blood and bronchoalveolar lavage . In contrast , aerosolized AERAS-402 alone or following BCG induced potent and stable Ag85A/b-specific P01730 and CD8 effector T cells in bronchoalveolar lavage that largely produced IFN-γ , as well as P01375 and P60568 . Such responses induced by BCG , AERAS-402 , or both failed to confer overall protection following challenge with 275 CFUs M. tuberculosis Erdman , although vaccine-induced responses associated with reduced pathology were observed in some animals . Anamnestic T cell responses to Ag85A/b were not detected in blood of immunized animals after challenge . Overall , our data suggest that a high M. tuberculosis challenge dose may be a critical factor in limiting vaccine efficacy in this model . However , the ability of aerosol rAd immunization to generate potent cellular immunity in the lung suggests that using different or more immunogens , alternative rAd serotypes with enhanced immunogenicity , and a physiological challenge dose may achieve protection against M. tuberculosis . [ Differential gene expression in nasopharyngeal carcinoma cell with reduced and normal expression of 6A8 alpha-mannosidase ] . OBJECTIVE : To detect the differential display of mRNA expression between human nasopharyngeal carcinoma cell CNE-2L2 with reduced malignancy caused by transduction of a DNA antisense to 6A8 alpha-mannosidase cDNA ( AS cell ) and the wild type cell ( W cell ) . METHODS : Differential display of mRNA expression was analyzed using DNA microarray analysis . The datasets were confirmed by Northern blotting and RT-PCR . RESULTS : Out of the 1069 genes analyzed , 34 genes were up-regulated in AS cells relative to W cells . Conversely , 42 genes were down-regulated . The genes , up-regulation of which might have suppressive effect on tumor malignant behaviors , were P130 mRNA for 130K protein , TGF-betaIIR alpha , Q9UBS5 , P36897 , Q13829 , STANIN , E-CADHERIN , P35221 and 2 , P48378 , TMPO , etc . The genes , down-regulation of which might have suppressive effect on tumor malignant behaviors , were P16070 , Q92597 , P01137 , P46782 , LEGUMAIIN , P35520 , P13987 , P09661 , etc . The microarray datasets were confirmed by Northern blot and RT-PCR analysis . CONCLUSIONS : In comparison to the W cell , AS cell has up-regulation of 34 genes and down-regulation of 42 genes . Changes of the gene expression may play a role in the malignancy reduction of AS cell . 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 . Inhibition of hemangiogenesis and lymphangiogenesis after normal-risk corneal transplantation by neutralizing P15692 promotes graft survival . PURPOSE : To evaluate the occurrence and time course of hem- and lymphangiogenesis after normal-risk corneal transplantation in the mouse model and to test whether pharmacologic strategies inhibiting both processes improve long-term graft survival . METHODS : Normal-risk allogeneic ( C57BL/6 to BALB/c ) and syngeneic ( BALB/c to BALB/c ) corneal transplantations were performed and occurrence and time course of hem- and lymphangiogenesis after keratoplasty was observed , by using double immunofluorescence of corneal flatmounts ( with CD31 as a panendothelial and Q9Y5Y7 as a lymphatic vascular endothelium-specific marker ) . A molecular trap designed to eliminate P15692 ( DB08885 (R1R2) ; 12.5 mg/kg ) was tested for its ability to inhibit both processes after keratoplasty and to promote long-term graft survival ( intraperitoneal injections on the day of surgery and 3 , 7 , and 14 days later ) . RESULTS : No blood or lymph vessels were detectable immediately after normal-risk transplantation in either donor or host cornea , but hem- and lymphangiogenesis were clearly visible at day 3 after transplantation . Both vessel types reached donor tissue at 1 week after allografting and similarly after syngeneic grafting . Early postoperative trapping of P15692 significantly reduced both hem- and lymphangiogenesis and significantly improved long-term graft survival ( 78 % vs. 40 % ; P < 0.05 ) . CONCLUSIONS : There is concurrent , P15692 -dependent hem- and lymphangiogenesis after normal-risk keratoplasty within the preoperatively avascular recipient bed . Inhibition of hem- and lymphangiogenesis ( afferent and efferent arm of an immune response ) after normal-risk corneal transplantation improves long-term graft survival , establishing early postoperative hem- and lymphangiogenesis as novel risk factors for graft rejection even in low-risk eyes . PARP and CHK inhibitors interact to cause DNA damage and cell death in mammary carcinoma cells . The present studies examined viability and DNA damage levels in mammary carcinoma cells following P09874 and O14757 inhibitor drug combination exposure . P09874 inhibitors [ AZD2281 ; ABT888 ; DB02690 ; AG014699 ] interacted with O14757 inhibitors [ P55089 -01 ; AZD7762 ; LY2603618 ] to kill mammary carcinoma cells . P09874 and O14757 inhibitors interacted to increase both single strand and double strand DNA breaks that correlated with increased γ P16104 phosphorylation . Treatment of cells with O14757 inhibitors increased the phosphorylation of O14757 and P27361 /2 . Knock down of Q13315 suppressed the drug-induced increases in O14757 and P27361 /2 phosphorylation and enhanced tumor cell killing by P09874 and O14757 inhibitors . Expression of dominant negative Q02750 enhanced drug-induced DNA damage whereas expression of activated Q02750 suppressed both the DNA damage response and tumor cell killing . Collectively our data demonstrate that P09874 and O14757 inhibitors interact to kill mammary carcinoma cells and that increased DNA damage is a surrogate marker for the response of cells to this drug combination . 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 . 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 . 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 . 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 . The kinase inhibitor staurosporine induces P55008 arrest at two points : effect on retinoblastoma protein phosphorylation and cyclin-dependent kinase 2 in normal and transformed cells . DB02010 ( ST ) , a protein kinase inhibitor , at a concentration of 20 nM arrests normal diploid fibroblasts 3 h into P55008 ( H. A. Crissman et al. , Proc. Natl. Acad. Sci. USA , 88 : 7580-7584 , 1991 ; K. Abe et al. , Exp. Cell Res. , 192 : 122-127 , 1991 ) . ST ( 2 nM ) induces a new P55008 arrest point at 6 h into P55008 . Partial phosphorylation of the retinoblastoma protein was observed at the 2 nM ST arrest point , whereas the retinoblastoma protein was unphosphorylated or underphosphorylated at the 20 nM arrest point . This correlated with the activity of the cyclin-dependent kinase 2 ( P24941 ) and the phosphorylation of the Thr160 residue of p33CDK2 . The cyclin E and cyclin D1/2 levels were reduced at the 20 nM ST arrest point . In HeLa cells that do not arrest in P55008 in response to 2 or 20 nM ST , the retinoblastoma protein and P24941 phosphorylations and P24941 activity were not affected by ST . These results suggest that ST inhibits one or more P55008 -regulating protein kinases , which lie upstream of P24941 . 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 . DB06186 in melanoma patients with brain metastasis : a retro-spective multicentre evaluation of thirty-eight patients . Treatment with ipilimumab , a monoclonal antibody that antagonizes cytotoxic T-lymphocyte antigen-4 ( P16410 ) , results in improved survival of patients with stage IIIc-IV melanoma . However , there is a lack of data on the efficacy of ipilimumab in patients with brain metastases . To evaluate the efficacy of ipilimumab for the treatment of brain metastasis in melanoma , a multicentre , retrospective analysis of 38 patients with brain metastases in melanoma , treated with ipilimumab in the context of the French Expanded Access Program , was performed . Three patients had a 3 partial response , 5 stable disease , 15 disease progression and 15 patients died during the induction phase due to disease progression . Median overall survival was 101 days ( range 54-154 ) . The brain metastases control rate was 16 % ( 6/38 ) . DB06186 may be effective in a few patients with central nervous system metastasis . However , patients with brain metastases and a low life expectancy may not benefit sufficiently from treatment with ipilimumab . 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 . 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 . 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 . A cytokine-controlled mechanism for integrated regulation of T-lymphocyte motility , adhesion and activation . The co-ordination of T-cell motility , adhesion and activation remains poorly understood . It is also unclear how these functions are co-ordinated with external stimuli . Here we unveil a series of molecular interactions in cis at the surface of T lymphocytes with potent effects on motility and adhesion in these cells , and communicating with proliferative responses . These interactions were controlled by the signature cytokines of T helper subsets interleukin-2 ( P60568 ) and P05112 . P01130 -related protein 1 ( Q07954 ) was found to play a key role for T-cell motility by promoting development of polarized cell shape and cell movement . Endogenous thrombospondin-1 ( P07996 -1 ) enhanced cell surface expression of Q07954 through Q08722 . Cell surface expressed Q07954 induced motility and processing of P07996 -1 while inhibiting adhesion to intercellular adhesion molecule 1 and fibronectin . P60568 , but not P05112 , stimulated synthesis of P07996 -1 and motility through P07996 -1 and Q07954 . Stimulation of the T-cell receptor ( TCR ) /CD3 complex inhibited P07996 -1 expression . Inhibitor studies indicated that Q07954 regulated P07996 -1 expression and promoted motility through JAK signalling . This Q07954 -mediated motogenic signalling was connected to Q08722 /Gi protein signalling and P60568 -induced signalling through P07996 -1 . The motogenic P07996 -1/ Q07954 mechanism antagonized TCR/CD3-induced T-cell proliferation . These results indicate that Q07954 in collaboration with P07996 -1 directs a counter-adhesive and counter-proliferative motogenic cascade . T cells seem programmed to prioritize movement before adhesion through this cascade . In conclusion , vital decision-making in T lymphocytes regulating motility , adhesive interactions and proliferation , are integrated through a molecular mechanism connecting different cell surface receptors and their signalling pathways .

MH_train_1405

MH_train_1405

MH_train_1405

interacts_with DB08815?

multiple_choice

8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . 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 . Organization and nucleotide sequence of a transcriptional unit of Methanococcus vannielii comprising genes for protein synthesis elongation factors and ribosomal proteins . By a chromosome walking strategy the DNA region from Methanococcus vannielii flanking the genes for protein synthesis elongation factor ( EF ) 1 alpha and P13639 was cloned and sequenced . A gene organization of 5' - beta ' - open reading frame ( ORF ) 1 - ORF2 - P28222 - S7 - P13639 - EF-1 alpha - S10 - ORF3 - ORF4 - 3' was found where beta ' , P28222 , S7 , S10 , P13639 , and EF-1 alpha represent gene products with sequences similar to the beta ' subunit of RNA polymerase , ribosomal proteins P28222 , S7 , and S10 , and EF-G and EF-Tu from Escherichia coli , respectively . ORF1-4 represent gene products with no known eubacterial counterparts . Northern blot analysis of transcripts and nuclease S1 mapping showed that transcription initiates between beta ' and ORF1 and terminates at the 3' side of the S10 gene and that the genes from ORF1 to S10 are cotranscribed . Apart from the presence of two additional ORFs , ORF1 and ORF2 , and of the gene for S10 , this organization is identical to that of the eubacterial " streptomycin operon. " ORF1 displays sequence similarity to rat liver ribosomal protein L30 and may represent one of the " additional " ribosomal proteins of Methanococcus . The sequenced part of the beta ' gene and the P13639 and EF-1 alpha gene products from Methanococcus are more similar to their eukaryotic than to their eubacterial counterparts . It appears , therefore , that the genetic organization of the translational components resembles the situation in eubacteria , whereas their primary structures are more eukaryotic in nature . Biological profile of oestrogen receptor positive primary breast cancers in the elderly and response to primary endocrine therapy . P11511 inhibitors have been shown to be superior to Tamoxifen in several settings . It is unclear whether this superiority extends to their use as primary endocrine therapy in elderly patients with early operable primary breast cancer . Biological characteristics of the tumours may aid in selecting the most suitable agent . Primary endocrine therapy with DB01217 in 64 women > 70 years with oestrogen receptor alpha-positive ( ERalpha+ ) breast cancer was compared to that in 84 treated with Tamoxifen during the same period . Biomarkers were assessed by immunohistochemistry on diagnostic core biopsies . There was no significant difference between the two groups ( DB01217 vs Tamoxifen ) in terms of clinical benefit rates at 6 months ( 97 % vs 100 % ) or median progression free survival ( 16.5 vs 22.5 months ) . There were no withdrawals due to adverse events from DB01217 , compared to four with Tamoxifen . 46 % , 99 % , 8 % and 5 % of all patients were positive for progesterone receptor , ERbeta2 , P04626 and P00533 , respectively , and 64 % of patients had a moderate Ki-67 index . Positive P04626 status ( 18 vs 21 months , p=0.003 ) and moderate Ki-67 index ( 17.5 vs 23 months , p=0.042 ) were associated with significantly shorter progression free survival . Results thus far show that primary endocrine therapy with DB01217 in elderly patients with early operable ERalpha+ breast cancer is similar to Tamoxifen in terms of efficacy , but appears to be associated with less adverse events leading to withdrawals . In this population , ERalpha+ breast cancer also appears to have a less aggressive biological profile favouring better hormone sensitivity . Effects of lurasidone on executive function in common marmosets . Cognitive impairment is one of the major symptoms of schizophrenia , and is considered largely due to dysfunctions in the prefrontal cortex ( P27918 ) . DB08815 , a novel atypical antipsychotic agent with high binding affinity for dopamine D2 , serotonin P34969 , 5- Q13049 and P08908 receptors has been reported to have superior efficacy in rodents ' models of cognitive impairment . However , the beneficial effect of lurasidone on cognitive impairment has not been evaluated in non-human primates . In this study , we investigated the effect of lurasidone on executive function , which is one of the cognitive domains , in common marmosets and compared the results to those of other antipsychotics . The effects of lurasidone , haloperidol , olanzapine , risperidone , quetiapine and clozapine on executive function were evaluated in naïve marmosets using the object retrieval with detours ( ORD ) task . Before drug treatment , marmosets ' success rates in the easy trial of the test were almost 90 % . However , maximum success in the difficult trial of the task reached only 50 % after 8 days of training . DB00502 , olanzapine and risperidone decreased correct performance even in the easy trial of the task . All drugs , except lurasidone , impaired success rate in the difficult trial . On the other hand , lurasidone dose-dependently increased marmosets ' success rates in the difficult trial with significant effect at 10mg/kg . In conclusion , we have shown in this study that lurasidone , unlike conventional antipsychotics , improves cognition associated with executive function in common marmosets . These findings suggest that lurasidone would be more useful for treatment of schizophrenia cognitive impairment than other antipsychotics . 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 . 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 . The diphthamide modification pathway from Saccharomyces cerevisiae -- revisited . DB03223 is a conserved modification in archaeal and eukaryal translation elongation factor 2 ( P13639 ) . Its name refers to the target function for diphtheria toxin , the disease-causing agent that , through ADP ribosylation of diphthamide , causes irreversible inactivation of P13639 and cell death . Although this clearly emphasizes a pathobiological role for diphthamide , its physiological function is unclear , and precisely why cells need P13639 to contain diphthamide is hardly understood . Nonetheless , the conservation of diphthamide biosynthesis together with syndromes ( i.e. ribosomal frame-shifting , embryonic lethality , neurodegeneration and cancer ) typical of mutant cells that can not make it strongly suggests that diphthamide-modified P13639 occupies an important and translation-related role in cell proliferation and development . Whether this is structural and/or regulatory remains to be seen . However , recent progress in dissecting the diphthamide gene network ( Q9BZG8 - Q9BTV6 ) from the budding yeast Saccharomyces cerevisiae has significantly advanced our understanding of the mechanisms required to initiate and complete diphthamide synthesis on P13639 . Here , we review recent developments in the field that not only have provided novel , previously overlooked and unexpected insights into the pathway and the biochemical players required for diphthamide synthesis but also are likely to foster innovative studies into the potential regulation of diphthamide , and importantly , its ill-defined biological role . 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 . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Q9BVG9 : high efficiency for synthesizing phosphatidylserine containing docosahexaenoic acid . DB00144 ( PS ) , the major anionic phospholipid in eukaryotic cell membranes , is synthesized by the integral membrane enzymes PS synthase 1 ( PSS1 ) and 2 ( Q9BVG9 ) . Q9BVG9 is highly expressed in specific tissues , such as brain and testis , where docosahexaenoic acid ( DB01708 , 22:6n-3 ) is also highly enriched . The purpose of this work was to characterize the hydrocarbon-chain preference of Q9BVG9 to gain insight on the specialized role of Q9BVG9 in PS accumulation in the DB01708 -abundant tissues . Flag-tagged Q9BVG9 was expressed in P29320 cells and immunopurified in a functionally active form . Purified Q9BVG9 utilized both PE plasmalogen and diacyl PE as substrates . Nevertheless , the latter was six times better utilized , indicating the importance of an ester linkage at the sn-1 position . Although no sn-1 fatty acyl preference was noted , Q9BVG9 exhibited significant preference toward DB01708 compared with 18:1 or 20:4 at the sn-2 position . Preferential production of DB01708 -containing PS ( DB01708 -PS ) was consistently observed with Q9BVG9 purified from a variety of cell lines as well as with microsomes from mutant cells in which PS synthesis relies primarily on Q9BVG9 . These findings suggest that Q9BVG9 may play a key role in PS accumulation in brain and testis through high activity toward DB01708 -containing substrates that are abundant in these tissues . Interleukin 5 regulation of peritoneal B-cell proliferation and antibody secretion . The influence of recombinant interleukin 5 ( rIL-5 ) on murine peritoneal B-cell proliferation and antibody secretion was examined . Larger , low buoyant density peritoneal B cells proliferated better with rIL-5 than the smaller resting B cells. this was also true for splenic B cells ; however , comparison of the respective populations showed the large peritoneal B-cell responses to be superior . Limiting dilution analyses showed that from 25 % to about 40 % of large peritoneal B cells proliferated in response to rIL-5 when lipopolysaccharide ( LPS ) was present . No detectable difference in the fraction of proliferating splenic B cells was seen in the presence of rIL-5 . These results are consistent with expression of P05113 receptors on about 70 % of low-density peritoneal B cells as determined by fluorescent staining with anti-Il-5 receptor monoclonal antibody ( MoAb ) . P05113 also enhanced spontaneous and mitogen-driven IgM secretion by both peritoneal and splenic B lymphocytes ; the increases exhibited by peritoneal B cells , however , were at least twice those exhibited by splenic B cells . Spontaneous and mitogen-driven secretion of auto-antibodies to bromelain-treated mouse erythrocytes ( BrMRBC ) by peritoneal B cells were also increased by this interleukin . Furthermore , rIL-5 enhanced peritoneal B-cell plaque-forming cell ( P27918 ) responses to TNP-LPS but not to TNP-Ficoll . Both an anti-IL-5R MoAb and an anti- P05113 MoAb blocked the rIL-5-induced enhancement of proliferation and auto-antibody P27918 responses . Hence , P05113 appears to be important for the regulation of proliferation and antibody secretion by many murine peritoneal B cells . DB06612 in eosinophilic disorders . DB06612 ( Bosatria(®) , GlaxoSmithKline ) is a biologic agent developed to treat asthma . It represents a humanized monoclonal antibody of IgG1 κ type , which targets human P05113 and thus prevents its interaction with the α-chain of the P05113 receptor . To date , it has not been approved for use in any eosinophil-related disorder ; however , several studies have suggested some therapeutic benefit across a spectrum of eosinophil-related disorders . This article evaluates the currently available preclinical and clinical studies , and the impact of mepolizumab against a variety of eosinophilic disorders . Prolonged opportunity for neuroprotection in experimental stroke with selective blockade of cyclooxygenase-2 activity . The post-treatment effects of the selective cyclooxygenase ( P36551 ) -2 inhibitor , valdecoxib , were investigated in a rat model of temporary focal ischemia . DB00580 reduced basal brain prostaglandin E(2) concentrations at dosages that did not affect serum thromboxane B(2) , consistent with a selective P35354 effect . Temporary focal cerebral ischemia was produced in rats by middle cerebral artery occlusion for 90 min . There was increased expression of P35354 protein detected by Western blot and immunocytochemistry within neurons in the ischemic cortex at 4 and 24 h after ischemia . Rats were treated with vehicle or valdecoxib 15 min before or 1.5 , 3 and 6 h after cerebral ischemia . Rats were sacrificed and brain infarction volume determined 24 h after ischemia . DB00580 treatment was associated with a decrease in infarction volume when administered 15 min before , and 1.5 or 3 h but not 6 h after cerebral ischemia . There were no differences in physiological parameters during the procedure . DB00580 administered at 1.5 h after ischemia significantly reduced the concentrations of prostaglandin E(2) in ischemic penumbral cortex as compared to the vehicle-treated group and contralateral non-ischemic cortex . These results suggest that P35354 inhibition with valdecoxib is effective when initiated both before and after middle cerebral artery occlusion . 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 . Ligand-dependent interactions of coactivators steroid receptor coactivator-1 and peroxisome proliferator-activated receptor binding protein with nuclear hormone receptors can be imaged in live cells and are required for transcription . Members of the nuclear receptor superfamily are thought to activate transcription by recruitment of one or more recently identified coactivator complexes . Here we demonstrate that both peroxisome proliferator-activated receptor binding protein ( PBP ) and steroid receptor coactivator-1 ( Q15788 ) are required for ligand-dependent transcription of transiently transfected and chromosomally integrated reporter genes by the estrogen receptor ( ER ) and retinoic acid receptor ( RAR ) . To examine ligand-dependent interactions between nuclear receptors and specific coactivators in living cells , these proteins were tagged with cyan ( P27918 ) and yellow ( YFP ) mutants of the green fluorescent protein . Fluorescence resonance energy transfer ( FRET ) from the P27918 to the YFP indicated interaction between the receptor and coactivator . P27918 fusions to RAR or its ligand-binding domain exhibited rapid ligand-dependent FRET to YFP-tagged nuclear receptor interaction domains of the coactivators Q15788 and PBP . The ER-ligand-binding domain , unlike RAR , also exhibited some basal interaction with coactivators in unstimulated cells that was abolished by the receptor antagonists tamoxifen or ICI182,780 . Inhibition of FRET by tamoxifen but not ICI182,780 could be reversed by estradiol , whereas estradiol-enhanced FRET could not be inhibited by either antagonist , indicating that ligand effects can show varying degrees of hysteresis . These findings suggest that ligand-dependent transcriptional activities of the RAR and ER require concurrent or sequential recruitment of Q15788 and PBP-containing coactivator complexes . DB01045 -independent interactions between the pregnane X receptor ligand binding domain and peptide fragments of coactivator and corepressor proteins . The pregnane X receptor ( O75469 ) , a member of the nuclear receptor superfamily , regulates the expression of drug-metabolizing enzymes in a ligand-dependent manner . The conventional view of nuclear receptor action is that ligand binding enhances the receptor 's affinity for coactivator proteins , while decreasing its affinity for corepressors . To date , however , no known rigorous biophysical studies have been conducted to investigate the interaction among O75469 , its coregulators , and ligands . In this work , steady-state total internal reflection fluorescence microscopy ( TIRFM ) and total internal reflection with fluorescence recovery after photobleaching were used to measure the thermodynamics and kinetics of the interaction between the O75469 ligand binding domain and a peptide fragment of the steroid receptor coactivator-1 ( Q15788 ) in the presence and absence of the established O75469 agonist , rifampicin . Equilibrium dissociation and dissociation rate constants of ~5 μM and ~2 s(-1) , respectively , were obtained in the presence and absence of rifampicin , indicating that the ligand does not enhance the affinity of the O75469 and Q15788 fragments . Additionally , TIRFM was used to examine the interaction between O75469 and a peptide fragment of the corepressor protein , the silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) . An equilibrium dissociation constant of ~70 μM was obtained for Q9Y618 in the presence and absence of rifampicin . These results strongly suggest that the mechanism of ligand-dependent activation in O75469 differs significantly from that seen in many other nuclear receptors . Pharmacokinetics of subcutaneously administered etanercept in subjects with psoriasis . AIMS : To present the results of the pharmacokinetic analysis of the concentration-time profiles of etanercept , a soluble receptor tumour necrosis factor ( P01375 ) antagonist , in more than 1300 subjects with psoriasis . METHODS : Pharmacokinetic samples were collected in one phase-2 and two phase-3 placebo-controlled , randomized clinical trials . Study 1 evaluated a 25-mg twice weekly ( BIW ) etanercept dosing regimen administered by subcutaneous ( s.c. ) injection for 24 weeks . Study 2 evaluated 25-mg BIW and 50-mg BIW s.c. doses for 12 weeks . Study 3 evaluated 25 mg once weekly ( QW ) , 25 mg BIW and 50 mg BIW s.c. doses for 24 weeks . RESULTS : The mean +/- SD steady-state predose serum concentrations of etanercept for the 25-mg BIW arm at 12 weeks in study 1 were 1590 +/- 885 ng ml(-1) . In study 2 , mean +/- SD etanercept steady-state concentrations at 12 weeks were 1900 +/- 1110 ng ml(-1) in the 25-mg BIW group and 3830 +/- 1870 ng ml(-1) in the 50-mg BIW group . The mean +/- SD steady-state predose serum concentrations of etanercept at 12 weeks in study 3 were 768 +/- 475 ng ml(-1) for the 25-mg QW regimen , 1990 +/- 1030 ng ml(-1) for the 25-mg BIW regimen and 4020 +/- 2100 ng ml(-1) for the 50-mg BIW regimen . CONCLUSIONS : Pharmacokinetic results were highly consistent across clinical trials . The concentration-time profiles displayed dose proportionality . DB00005 concentrations in subjects with psoriasis are similar to the concentrations in subjects with rheumatoid arthritis . 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 . 27-hydroxycholesterol is an endogenous ligand for liver X receptor in cholesterol-loaded cells . The nuclear receptors liver X receptor alpha ( LXRalpha ) ( Q13133 ) and LXRbeta ( P55055 ) are important regulators of genes involved in lipid metabolism , including O95477 , P45844 , and sterol regulatory element-binding protein-1c ( SREBP-1c ) . Although it has been demonstrated that oxysterols are LXR ligands , little is known about the identity of the physiological activators of these receptors . Here we confirm earlier studies demonstrating a dose-dependent induction of O95477 and P45844 in human monocyte-derived macrophages by cholesterol loading . In addition , we show that formation of 27-hydroxycholesterol and cholestenoic acid , products of Q02318 action on cholesterol , is dependent on the dose of cholesterol used to load the cells . Other proposed LXR ligands , including 20(S)-hydroxycholesterol , 22(R)-hydroxycholesterol , and 24(S),25-epoxycholesterol , could not be detected under these conditions . A role for Q02318 in regulation of cholesterol-induced genes was demonstrated by the following findings . 1 ) Introduction of Q02318 into P29320 -293 cells conferred an induction of P45844 and SREBP-1c ; 2 ) upon cholesterol loading , Q02318 -expressing cells induce these genes to a greater extent than in control cells ; 3 ) in Q02318 -deficient human skin fibroblasts , the induction of O95477 in response to cholesterol loading was ablated ; and 4 ) in a coactivator association assay , 27-hydroxycholesterol functionally activated LXR . We conclude that 27-hydroxylation of cholesterol is an important pathway for LXR activation in response to cholesterol overload .

MH_train_1406

MH_train_1406

MH_train_1406

interacts_with DB06144?

multiple_choice

Molecular targets and regulators of cardiac hypertrophy . Cardiac hypertrophy is one of the main ways in which cardiomyocytes respond to mechanical and neurohormonal stimuli . It enables myocytes to increase their work output , which improves cardiac pump function . Although cardiac hypertrophy may initially represent an adaptive response of the myocardium , ultimately , it often progresses to ventricular dilatation and heart failure which is one of the leading causes of mortality in the western world . A number of signaling modulators that influence gene expression , apoptosis , cytokine release and growth factor signaling , etc. are known to regulate heart . By using genetic and cellular models of cardiac hypertrophy it has been proved that pathological hypertrophy can be prevented or reversed . This finding has promoted an enormous drive to identify novel and specific regulators of hypertrophy . In this review , we have discussed the various molecular signal transduction pathways and the regulators of hypertrophic response which includes calcineurin , cGMP , NFAT , natriuretic peptides , histone deacetylase , P05231 cytokine family , Gq/ P49842 signaling , PI3K , MAPK pathways , Na/H exchanger , DB01367 , polypeptide growth factors , P01160 , NO , P01375 , Q07869 and JAK/ P35610 pathway , microRNA , Cardiac angiogenesis and gene mutations in adult heart . Augmented knowledge of these signaling pathways and their interactions may potentially be translated into pharmacological therapies for the treatment of various cardiac diseases that are adversely affected by hypertrophy . The purpose of this review is to provide the current knowledge about the molecular pathogenesis of cardiac hypertrophy , with special emphasis on novel researches and investigations . Characterization of the human gonadotropin-releasing hormone receptor heterologously produced using the baculovirus/insect cell and the Semliki Forest virus systems . 1. Two eukaryotic viral systems , the baculovirus/insect cell and the Semliki Forest virus systems , were tested for heterologous expression of human gonadotropin-releasing hormone receptor ( GnRHR ) cDNA . 2 . An unmodified as well as a c-myc epitope-tagged human P30968 was produced in two insect cell lines ( Spodoptera frugiperda , Trichoplusia ni ) after infection with the respective recombinant baculoviruses . In both insect cell lines , the receptor was identified by immunoblot analysis as a triplet of bands between 35 and 40 kDa . After deglycosylation of the receptor the molecular mass decreased to 35 kDa . The P30968 was localized in membrane compartments within the infected insect cells . However , only in membranes of infected Trichoplusia ni insect cells could approximately 2000 receptors per cell be detected . 3 . Production of the P30968 in BHK cells using the Semliki Forest virus system resulted in approximately 50,000 receptors per cell . A maximal yield of 0.42 pmol/mg membrane protein was obtained 24 hr after electroporation of BHK cells with in vitro synthesized RNA . Binding of the antagonist [125I] DB00050 was saturable with a KD of 1.3 nM . The receptor produced in the BHK cells was further characterized by ligand displacement studies . The rank order of agonist and antagonist affinities was DB00050 > DB06825 > Antide > DB00644 . Production of an antiserum specific to the ADP-ribosylated form of elongation factor 2 from archaebacteria and eukaryotes . An antiserum to ADP-ribosylated elongation factor 2 ( DB02059 - P13639 ) from S. acidocaldarius was raised in rabbits using stained , homogenized , DB02059 - P13639 -containing slices from SDS-gels as a source of antigen . P13639 ( P13639 ) from S. acidocaldarius was cloned in E. coli and the expressed gene product was used in order to adsorb all anti- P13639 antibodies which do not contain the ADP-ribosyl group within their epitopes , as E. coli is unable to synthesize the ADP-ribosyl acceptor diphthamide . The remaining antibodies were specific to ADP-ribosylated P13639 from Thermoplasma acidophilum , S. acidocaldarius and Desulfurococcus mucosus . ADP-ribosylated P13639 from eukaryotic sources also reacted with the adsorbed antiserum as shown for P13639 isolated from the killi-fish Cynolebias whitei , the mouse species BALB/c and Han/Wistar rats . The adsorbed antiserum did not cross-react with ADP-ribosylated actin or rho protein or with DB03147 -containing D-amino acid oxidase . Physiologically Based Pharmacokinetic Model to Assess the Influence of DB09052 -Mediated Cytokine Elevations on Cytochrome P450 Enzyme Activity . DB09052 is a P15391 /CD3 bispecific T-cell engager ( BiTE® ) antibody construct for treatment of leukemia . Transient elevation of cytokines ( interleukin ( IL ) -6 , P22301 , interferon-gamma ( IFN-γ ) ) has been observed within the first 48 hours of continuous intravenous blinatumomab infusion . In human hepatocytes , blinatumomab showed no effect on cytochrome P450 ( CYP450 ) activities , whereas a cytokine cocktail showed suppression of P08684 , P05177 , and P11712 activities . We developed a physiologically based pharmacokinetic ( PBPK ) model to evaluate the effect of transient elevation of cytokines , particularly P05231 , on CYP450 suppression . The predicted suppression of hepatic CYP450 activities was < 30 % , and P05231 -mediated changes in exposure to sensitive substrates of P08684 , P05177 , and P11712 were < twofold and lasted < 1 week . Model verification indicated that P05231 was the key cytokine suppressing CYP450 activities ; the duration of cytokine elevation was a major determinant of magnitude of suppression . This study shows the utility of PBPK modeling for risk assessment of cytokine-mediated drug interactions . Mitochondrial dysfunction promotes and aggravates the inflammatory response in normal human synoviocytes . OBJECTIVES : In RA , synoviocytes cause increased oxidative stress , leading to mitochondrial alterations that may participate in the pathogenesis of RA . Here we investigated whether mitochondrial dysfunction induces inflammatory responses in cultured normal human synoviocytes , a hallmark of RA . METHODS : Mitochondrial dysfunction was induced with the inhibitor oligomycin . The effects of mitochondrial dysfunction on cyclooxygenase-2 ( P35354 ) , prostaglandin E2 ( DB00917 ) and P10145 expression ; cellular and mitochondrial reactive oxygen species ( ROS ) production ; nuclear factor-κB ( NF-κB ) activation and p65 translocation were studied . ROS scavengers ( DB06151 and mitoTEMPO ) and an NF-κB inhibitor ( BAY-117085 ) were used to investigate the pathways involved . The natural anti-inflammatory antioxidant resveratrol was also tested . RESULTS : Mitochondrial dysfunction per se significantly stimulated mitochondrial ROS production as well as low-grade expressions of P35354 , DB00917 and P10145 . Interestingly , mitochondrial dysfunction induced by pretreatment of synoviocytes with oligomycin synergized with IL-1β to increase the expression of these inflammatory mediators . The inflammatory effects of mitochondrial damage appeared to be dependent on ROS production and NF-κB activation since the inflammatory response was counteracted by both DB06151 and mitoTEMPO and it was also reduced by BAY-117085 . Antimycin A and paraquat ( inhibitors of mitochondrial function ) also induced inflammatory responses . Furthermore , resveratrol significantly reduced the inflammatory response by decreasing ROS production and NF-κB activation . CONCLUSION : These data suggest that mitochondrial dysfunction could induce an inflammatory response in normal human synoviocytes and sensitize these cells , causing a significant amplification of the inflammatory response induced by IL-1β . DB02709 may represent a promising strategy in controlling the synovial inflammatory response . Ontogeny of gene expression in the gonadotroph of the developing female rat . During the infantile period of the female rat ( 8-21 postnatal days [ P01160 ] of age ) , there is a dramatic increase in plasma DB00094 , which is thought to be important in initiating ovarian activity and , perhaps , the onset of puberty . To begin to understand the regulation of this DB00094 surge , we determined the ontogenetic development of LHbeta , FSHbeta , and P30968 ( P30968 ) mRNA levels in the pituitary gland throughout the infantile period of the female rat . Steady-state mRNA levels were determined by an external standard quantitative reverse transcriptase polymerase chain reaction assay . FSHbeta and P30968 mRNA levels increased to peak on P01160 12 ( p < 0.03 ) . LHbeta mRNA levels remained relatively constant until rising on P01160 18 . A DB00644 antagonist ( 10-100 microg/animal ) was administered daily from P01160 8-11 or P01160 11-13 , and animals were killed on P01160 12 or P01160 14 , respectively . FSHbeta , LHbeta , and P30968 mRNAs were not affected by DB00644 antagonist treatment . Plasma DB00094 was selectively reduced in the first group , whereas both plasma LH and DB00094 were suppressed in the second group . These data indicate that gene expression of LHbeta , FSHbeta , and P30968 are differentially regulated in the infantile female rat pituitary . DB00644 is involved in regulating the secretion of DB00094 and LH during the infantile period but not in regulating FSHbeta , LHbeta , or P30968 mRNA gene expression . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . Q96EB6 , a class III histone deacetylase , regulates P01375 -α-induced inflammation in human chondrocytes . OBJECTIVE : The present study was performed to elucidate the possible role of Q96EB6 signaling in joint inflammation in human articular chondrocytes . DESIGN : Real-time quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) and Western blotting were performed to detect gene products and proteins involved in tumor necrosis factor α ( P01375 -α ) -induced inflammation and cartilage degradation in human primary chondrocytes . Matrix metalloproteinase ( MMP ) -2 and P14780 activity was evaluated by gelatin zymography . Overexpression and knockdown of Q96EB6 were also performed to investigate whether Q96EB6 is associated with the anti-inflammatory activity of resveratrol in chondrocytes . RESULTS : DB02709 dose-dependently inhibited P01375 -α-induced cyclooxygenase-2 ( P35354 ) , P03956 , P08254 , P45452 and PGE(2) production in human chondrocytes . Moreover , P08253 and P14780 activity was increased by treatment with P01375 -α ; however , Q96EB6 activation decreased the proinflammatory effects induced by P01375 -α . In addition , treatment of Q96EB6 activator and overexpression of Q96EB6 inhibited the expression and activation of the main proinflammatory regulator NF-κB , which was increased by P01375 -α . When Q96EB6 was overexpressed in chondrocytes , the anti-inflammatory action of Q96EB6 was similar to that exerted by resveratrol . CONCLUSIONS : Q96EB6 activation deacetylates and inactivates NF-κB , and thereby , exerts an anti-inflammatory effect on chondrocytes , suggesting that Q96EB6 activators could be explored as potential treatments for arthritis . 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 . Chemoimmunotherapy in acute lymphoblastic leukemia . ALL blast cells express a variety of specific antigens e.g. P15391 , P11836 , P20273 , P20138 , and P31358 , which serve as targets for Monoclonal Antibodies ( MoAbs ) . So far , the most experience is available for anti- P11836 ( rituximab ) , which has been combined with chemotherapy for treatment of mature B-ALL/Burkitt 's lymphoma . Studies with rituximab have also been completed in B-precursor ALL . Another antigen , P15391 , is of great interest due to a very high rate of expression in ALL . It can be targeted by a bispecific monoclonal antibody , DB09052 , directed against P15391 and CD3 . Smaller studies or case reports are also available for the anti P31358 antibody ( DB00087 ) , for anti P20273 ( DB04958 ) or anti P20138 ( Gemtuzumab ) . Available data demonstrate that MoAb therapy in ALL is a highly promising targeted treatment . However , several details for an optimal treatment approach e.g. the required level of antigen expression , timing , schedule , dosage and stage of disease still need to be defined . Impact of aromatic residues within transmembrane helix 6 of the human gonadotropin-releasing hormone receptor upon agonist and antagonist binding . To investigate the impact of aromatic residues within transmembrane helix 6 ( TMH6 ) of the human gonadotropin-releasing hormone receptor ( P30968 ) on agonist and antagonist binding , residues Y(283) , Y(284) , W(289) , Y(290) , W(291) , and F(292) were exchanged to alanine and analyzed comprehensively in functional reporter gene and ligand binding assays . Whereas receptor mutants Y(283)A , Y(284)A , and W(291)A were capable of neither ligand binding nor signal transduction , mutants W(289)A , Y(290)A , and F(292)A were functional : the F(292)A mutant behaved like wild-type receptor , while mutants W(289)A and Y(290)A differentiated between agonistic and antagonistic ligands . On the basis of the high-resolution X-ray structure of bovine rhodopsin as well as available data on P30968 mutants , models for ligand-receptor interactions are proposed . The model for D- DB00150 (6)- DB00644 ( DB06825 ) binding , representing a superagonistic ligand , is in full accordance to available data . Furthermore , new interactions are proposed : pGlu(1) interacts with N(212) in transmembrane helix 5 , DB00135 (5) with Y(290) , and D- DB00150 (6) with W(289) . The binding behavior of mutants W(289)A and Y(290)A corresponds to the proposed binding model for the antagonist DB00050 . In summary , our data as presented indicate that Y(290) plays a key function in agonist but not antagonist binding . 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 . DB08875 ( DB05153 ) for the treatment of locally advanced or metastatic progressive medullary thyroid cancer . DB08875 ( DB05153 ) is an oral multiple receptor tyrosine kinase inhibitor manufactured by Exelixis Inc. , CA , USA . It mainly inhibits three tyrosine kinase receptors : MET , P35968 and P07949 . In both preclinical and clinical studies it has been shown to inhibit tumor angiogenesis , invasiveness and metastases . The most frequent side effects are fatigue , diarrhea , decreased appetite , nausea , weight loss and palmar-plantar erythrodysesthesia . A Phase III clinical trial ( EXAM study ) of DB05153 versus placebo in advanced and progressive medullary thyroid cancer showed a 28 versus 0 % overall response rate and a progression-free survival of 11.2 versus 4.0 months ( hazard ratio : 0.28 ; 95 % CI : 0.19-0.40 ; p < 0.0001 ) in patients treated with cabozantinib and placebo , respectively . The drug has been approved by the US FDA for the treatment of advanced/progressive metastatic medullary thyroid cancer in the USA . The P15941 is now evaluating its approval in Europe . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . 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 . An improved effect size for single-case research : nonoverlap of all pairs . Nonoverlap of All Pairs ( Q8WYA6 ) , an index of data overlap between phases in single-case research , is demonstrated and field tested with 200 published AB contrasts . Q8WYA6 is a novel application of an established effect size known in various forms as Area Under the Curve ( AUC ) , the Common Language Effect Size ( CL ) , the Probability of Superiority ( PS ) , the Dominance Statistic ( DS ) , Mann-Whitney 's U , and Sommers D , among others . Q8WYA6 was compared with 3 other non-overlap-based indices : P01160 ( percent of nonoverlapping data ) , P15941 ( percent of data points exceeding the median ) , and PAND ( percent of all nonoverlapping data ) , as well as Pearson 's R(2) . Five questions were addressed about Q8WYA6 : ( a ) typical Q8WYA6 values , ( b ) its ability to discriminate among typical single-case research results , ( c ) its power and precision ( confidence interval width ) , ( d ) its correlation with the established effect size index , R(2) , and ( e ) its relationship with visual judgments . Results were positive , the new index equaling or outperforming the other overlap indices on most criteria . Celecoxib blocks cardiac Kv1.5 , Kv4.3 and Kv7.1 ( P51787 ) channels : effects on cardiac action potentials . Celecoxib is a P35354 inhibitor that has been related to an increased cardiovascular risk and that exerts several actions on different targets . The aim of this study was to analyze the effects of this drug on human cardiac voltage-gated potassium channels ( Kv ) involved on cardiac repolarization Kv1.5 ( I(Kur) ) , Kv4.3+ Q9NS61 ( I(to1) ) and Kv7.1+ P15382 ( I(Ks) ) and to compare with another P35354 inhibitor , rofecoxib . Currents were recorded in transfected mammalian cells by whole-cell patch-clamp . Celecoxib blocked all the Kv channels analyzed and rofecoxib was always less potent , except on Kv4.3+ Q9NS61 channels . Kv1.5 block increased in the voltage range of channel activation , decreasing at potentials positive to 0 mV . The drug modified the activation curve of the channels that became biphasic . Block was frequency-dependent , increasing at fastest frequencies . Celecoxib effects were not altered by DB08837 (out) in R487Y mutant Kv1.5 channels but the kinetics of block were slower and the degree of block was smaller with DB08837 (in) , indicating that celecoxib acts from the cytosolic side . We confirmed the blocking properties of celecoxib on native Kv currents from rat vascular cells , where Kv1.5 are the main contributors ( IC(50)≈ 7 μM ) . Finally , we demonstrate that celecoxib prolongs the action potential duration in mouse cardiac myocytes and shortens it in guinea pig cardiac myocytes , suggesting that Kv block induced by celecoxib may be of clinical relevance . 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 . Expression of two variants of the human mu opioid receptor mRNA in SK-N-SH cells and human brain . A partial mu opioid receptor gene was isolated from a human genomic library using a mouse delta opioid receptor cDNA as a probe . Using information from this genomic clone and the published human mu receptor , P35372 , a cDNA was isolated from SK-N-SH mRNA that codes for a variant of the P35372 mRNA , MOR1A . The presence of MOR1A is also shown in human brain using RT-PCR . MOR1A differs from P35372 in that the 3' terminal intron has not been removed . An in-frame termination codon is found four amino acids after the 5' consensus splice site , making MOR1A eight amino acids shorter than P35372 . Both receptors show similar ligand binding and coupling to DB02527 in CHO- P04264 cells . The C-terminal differences between P35372 and MOR1A could have effects on receptor coupling or receptor transport and localization . Irritable bowel syndrome neuropharmacology . A review of approved and investigational compounds . Anticholinergics and prokinetics are mainstays of therapy for Irritable Bowel Syndrome ( IBS ) patients despite their limited efficacy and troublesome side-effect profile . The clinical limitations of these drugs are a result of their relative broad and nonspecific pharmacologic interaction with various receptors . Recent advances in gut physiology have led to the identification of various receptor targets that may play a pivotal role in the pathogenesis of IBS . Medicinal chemists searching for safe and effective IBS therapies are now developing compounds targeting many of these specific receptors . The latest generation of anticholinergics , such as zamifenacin , darifenacin , and YM-905 , provide selective antagonism of the muscarinic type-3 receptor . DB01079 , a selective Q13639 partial agonist , tested in multiple clinical trials , is effective in reducing the symptoms of abdominal pain , bloating , and constipation . Ezlopitant and nepadudant , selective antagonists for neurokinin receptors type 1 and type 2 , respectively , show promise in reducing gut motility and pain . DB00836 , a mu ( mu ) opioid receptor agonist , is safe and effective for IBS patients with diarrhea ( IBS-D ) as the predominant bowel syndrome . Fedotozine , a kappa ( kappa ) opioid receptor agonist , has been tried as a visccral analgesic in various clinical trials with conflicting results . DB00969 , a 5- Q9H205 receptor antagonist , has demonstrated efficacy in IBS-D patients but incidents of ischemic colitis seen in post-marketing follow-up resulted its removal from the market . Compounds that target cholecystokinin . A , N-methyl-D-aspartate , alpha 2-adrenergic , and DB05394 receptors are also examined in this review . 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 . 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 . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 ) -deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5-hydroxy-l-tryptophan ( 5-HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . A common single nucleotide polymorphism can exacerbate long-QT type 2 syndrome leading to sudden infant death . BACKGROUND : Identification of infants at risk for sudden arrhythmic death remains one of the leading challenges of modern medicine . We present a family in which a common polymorphism ( single nucleotide polymorphism ) inherited from the father , combined with a stop codon mutation inherited from the mother ( both asymptomatic ) , led to 2 cases of sudden infant death . METHODS AND RESULTS : P51787 , Q12809 , Q14524 , P15382 , Q9Y6J6 , CACNA1c , CACNB2b , and P63252 genes were amplified and analyzed by direct sequencing . Functional electrophysiological studies were performed with the single nucleotide polymorphism and mutation expressed singly and in combination in Chinese ovary ( CHO- P04264 ) and COS-1 cells . An asymptomatic woman presenting after the death of her 2-day-old infant and spontaneous abortion of a second baby in the first trimester was referred for genetic analysis . The newborn infant had nearly incessant ventricular tachycardia while in utero and a prolonged QTc ( 560 ms ) . The mother was asymptomatic but displayed a prolonged QTc . Genetic screening of the mother revealed a heterozygous nonsense mutation ( P926AfsX14 ) in Q12809 , predicting a stop codon . The father was asymptomatic with a normal QTc but had a heterozygous polymorphism ( K897T ) in Q12809 . The baby who died at 2 days of age and the aborted fetus inherited both K897T and P926AfsX14 . Heterologous coexpression of K897T and P926AfsX14 led to loss of function of Q12809 current much greater than expression of K897T or P926AfsX14 alone . CONCLUSIONS : Our data suggest that a common polymorphism ( K897T ) can markedly accentuate the loss of function of mildly defective Q12809 channels , leading to long-QT syndrome-mediated arrhythmias and sudden infant death . Tp53-associated growth arrest and DNA damage repair gene expression is attenuated in mammary epithelial cells of rats fed whey proteins . Dietary protection from mammary cancer is likely coordinated through multiple signaling pathways , based on the known heterogeneity of the disease and the distinct origins of mammary tumor cells . The present study examined the modulatory effects of dietary intake of whey protein hydrolysate ( WPH ) relative to casein ( CAS ) , on mammary epithelial cell resistance to endogenous DNA damage using Tp53 gene expression and signaling as a read-out , and on systemic proapoptotic and immune surveillance activity , in young adult female Sprague-Dawley rats . Rats were fed AIN-93G diets made with CAS or WPH as the sole protein source beginning at gestation d 4 . At postnatal day ( P01160 ) 50 , mammary glands of rats fed WPH had lower levels of activated Tp53 and p38 mitogen-activated protein kinase proteins , and reduced transcript levels for Tp53-associated DNA damage repair , growth arrest , and proapoptotic genes than those of CAS-fed rats . Serum from WPH-fed rats had greater apoptotic activity in MCF-7 tumor cells than that from rats fed CAS . Serum levels of monocyte chemoattractant protein ( MCP ) -1 were higher in WPH- than in CAS-fed rats . MCF-7 cells treated with CAS serum + recombinant rat P13500 had apoptotic activity and Tp53 and P38936 gene expression levels comparable to those treated with WPH serum or recombinant P13500 . Results indicate that mammary glands of rats fed a WPH diet are more protected from endogenous DNA damage than are those of CAS-fed rats , and identify P13500 as a potential serum biomarker for the positive effects of healthy diets . 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 . 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 . DB01373 -induced folding of a fragment of calmodulin composed of EF-hands 2 and 3 . P62158 ( P62158 ) is an EF-hand protein composed of two calcium ( Ca(2+) ) -binding EF-hand motifs in its N-domain ( EF-1 and P13639 ) and two in its C-domain ( EF-3 and Q8N442 ) . In this study , we examined the structure , dynamics , and Ca(2+)-binding properties of a fragment of P62158 containing only P13639 and EF-3 and the intervening linker sequence ( CaM2/3 ) . Based on NMR spectroscopic analyses , Ca(2+)-free CaM2/3 is predominantly unfolded , but upon binding Ca(2+) , adopts a monomeric structure composed of two EF-hand motifs bridged by a short antiparallel beta-sheet . Despite having an " even-odd " pairing of EF-hands , the tertiary structure of CaM2/3 is similar to both the " odd-even " paired N- and C-domains of Ca(2+)-ligated P62158 , with the conformationally flexible linker sequence adopting the role of an inter-EF-hand loop . However , unlike either P62158 domain , CaM2/3 exhibits stepwise Ca(2+) binding with a K ( d1 ) = 30 +/- 5 microM to EF-3 , and a K ( d2 ) > 1000 microM to P13639 . Binding of the first equivalent of Ca(2+) induces the cooperative folding of CaM2/3 . In the case of native P62158 , stacking interactions between four conserved aromatic residues help to hold the first and fourth helices of each EF-hand domain together , while the loop between EF-hands covalently tethers the second and third helices . In contrast , these aromatic residues lie along the second and third helices of CaM2/3 , and thus are positioned adjacent to the loop between its " even-odd " paired EF-hands . This nonnative hydrophobic core packing may contribute to the weak Ca(2+) affinity exhibited by P13639 in the context of CaM2/3 . Chronic atrial fibrillation alters the functional properties of If in the human atrium . INTRODUCTION : Despite the evidence that the hyperpolarization-activated current ( If ) is highly modulated in human cardiomyopathies , no definite data exist in chronic atrial fibrillation ( cAF ) . We investigated the expression , function , and modulation of If in human cAF . METHODS AND RESULTS : Right atrial samples were obtained from sinus rhythm ( SR , n = 49 ) or cAF ( duration > 1 year , n = 31 ) patients undergoing corrective cardiac surgery . Among f-channel isoforms expressed in the human atrium ( O60741 , 2 and 4 ) , Q9Y3Q4 mRNA levels measured by RT-PCR were significantly reduced . However , protein expression was preserved in cAF compared to SR ( +85 % for Q9Y3Q4 ) ; concurrently , miR-1 expression was significantly reduced . In patch-clamped atrial myocytes , current-specific conductance ( gf ) was significantly increased in cAF at voltages around the threshold for If activation ( -60 to -80 mV ) ; accordingly , a 10-mV rightward shift of the activation curve occurred ( P < 0.01 ) . β-Adrenergic and Q13639 receptor stimulation exerted similar effects on If in cAF and SR cells , while the P01160 -mediated effect was significantly reduced ( P < 0.02 ) , suggesting downregulation of natriuretic peptide signaling . CONCLUSIONS : In human cAF modifications in transcriptional and posttranscriptional mechanisms of HCN channels occur , associated with a slight yet significant gain-of-function of If , which may contribute to enhanced atrial ectopy . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . DB01079 for constipation-predominant irritable bowel syndrome . DB01079 , a selective and partial agonist at the 5-hydroxytryptamine ( 5-HT [ serotonin ] ) receptor subtype 4 ( Q13639 ) , is the only United States Food and Drug Administration-approved drug for the treatment of constipation-predominant irritable bowel syndrome ( IBS ) in women . The drug 's stimulation of Q13639 receptors on intestinal enterocytes increases peristaltic activity and fluid secretion into the gut lumen , facilitating stool passage . In addition , affinity of tegaserod for Q13639 receptors modulates visceral sensitivity , which helps alleviate abdominal pain associated with constipation-predominant IBS . The drug 's pharmacokinetic and pharmacodynamic parameters do not differ significantly with age or sex . DB01079 safely and effectively relieves overall gastrointestinal symptoms and abdominal discomfort and normalizes bowel habits in patients with constipation-predominant IBS . It is associated with few drug interactions . In clinical studies , tegaserod was well tolerated , and its adverse-effect profile was similar to that of placebo . Severe diarrhea , as well as abdominal pain , flatulence , headache , and nausea , were the most commonly reported events . Patients who experience severe diarrhea should discontinue the drug . With the data available , tegaserod remains an option for patients with constipation-predominant IBS . Changes in expression of DB01221 receptor subunit mRNA by perinatal exposure to dioxin . Since dioxin and related compounds are suspected of affecting permanently the brain function of offspring of human and experimental animals , effects of perinatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin ( TCDD ) on the expression of rat DB01221 receptor Q12879 and Q13224 subunit mRNA were examined . The mRNA quantification by competitive RT-PCR clearly revealed that TCDD inhibited Q13224 mRNA expression and enhanced Q12879 mRNA expression in the neocortex and hippocampus on postnatal day ( P01160 ) 49 , whereas these changes in mRNA expression were not found on P01160 5 . The results demonstrate for the first time that the perinatal exposure to TCDD can alter the molecular basis of brain of offspring in adulthood . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Altered gene expression by low-dose arsenic exposure in humans and cultured cardiomyocytes : assessment by real-time PCR arrays . Chronic arsenic exposure results in higher risk of skin , lung , and bladder cancer , as well as cardiovascular disease and diabetes . The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array ( TLDA ) . We found that expression of tumor necrosis factor-α ( P01375 -α ) , which activates both inflammation and NF-κB-dependent survival pathways , was strongly associated with water and urinary arsenic levels . Expression of P22460 , which encodes a potassium ion channel protein , was positively associated with water and toe nail arsenic levels . Expression of 2 and 11 genes were positively associated with nail and urinary arsenic , respectively . Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans , we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro . Expression of the ion-channel genes CACNA1 , Q12809 , P51787 and P15382 were down-regulated by 1-μM arsenic . Alteration of some common pathways , including those involved in oxidative stress , inflammatory signaling , and ion-channel function , may underlay the seemingly disparate array of arsenic-associated diseases , such as cancer , cardiovascular disease , and diabetes . 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 . Atrial natriuretic peptide : a possible mediator involved in dexamethasone 's inhibition of cell proliferation in multiple myeloma . Atrial natriuretic peptide ( P01160 ) has been recognized for several decades for its role of regulating blood pressure . Recently , cumulating evidences show that P01160 plays an anticancer role in various solid tumors via blocking the kinase cascade of Ras- Q02750 /2- P27361 /2 with the result of inhibition of DNA synthesis . P01160 , as well as its receptors ( P16066 and P17342 ) has been identified present in the embryonic stem cell and a wide range of cancer cells . Various lymphoid organs , such as lymph nodes , have been detected the presence of P01160 . Multiple myeloma ( MM ) , though the therapies have evolved significantly , is still an incurable disease as B lymphocyte cell neoplasm . Dexamethasone is the cornerstone in treatment of MM via inactivation of Ras- Q02750 /2- P27361 /2 cascade reaction . Coincidently , dexamethasone can increase the expression of P01160 markedly . Nevertheless , the role of P01160 in MM is unclear . Based on these results above , we raise the hypothesis that P01160 is involved in mediating dexamethasone 's inhibition of proliferation in MM cells , which suggests that P01160 may be a potential agent to treat MM .

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DB01296 sulfate suppresses the expressions of urokinase plasminogen activator and inhibitor and gelatinases during the early stage of osteoarthritis . BACKGROUND : DB01296 sulfate may have an ex vivo inhibitory effect on the plasminogen activator ( PA ) /plasmin system and gelatinases expression during the early development of osteoarthritis ( OA ) . METHODS : We compared the levels of urokinase-type PA ( u-PA ) , PA inhibitor-1 ( P05121 ) and gelatinases ( matrix metalloproteinase-2 and -9 [ P08253 and -9 ] ) in a series of chondral , meniscal , and synovial cultures of early OA after treatment with or without glucosamine sulfate . RESULTS : Gelatin zymography revealed that glucosamine sulfate could suppress P08253 secretion in chondral , meniscal and synovial cultures and also decrease P14780 production in synovial and meniscal cultures . ELISA data also showed the suppressive effects of glucosamine sulfate on u-PA and P05121 production in synovial cultures at 48 h . CONCLUSIONS : Our data suggest that one of the therapeutic effects of glucosamine sulfate is to down-regulate the expressions of u-PA , P05121 , P08253 and P14780 that underlie the destruction of articular cartilage in the early stage of OA , and therefore to delay the joint failure . Q9H244 receptors play a significant role in the development of platelet microaggregation in patients with diabetes . Ninety-eight diabetic patients ( type 2 ) were studied together with 24 healthy normotensive controls . Microaggregates ( particle scale , < 25 microm ) of platelets were detected by a laser scattering system . Microaggregates in the control group showed a time-dependent reversible change ; however , they existed continuously in 82 of 98 diabetic patients . When platelets of diabetics were stimulated by a shear stress alone without ADP , 74 also showed spontaneous and irreversible microaggregates even though they were not observed in all control subjects . In control subjects , microaggregates were inhibited by MRS2279 ( a P47900 antagonist ) , but not AR-C69931MX ( a Q9H244 antagonist ) . However , AR-C69931MX prevented irreversible microaggregates in diabetic patients . When either aspirin or ticlopidine was administered to diabetic patients with irreversible microaggregates , both drugs significantly decreased microaggregates induced by a low dose of ADP . DB00208 additionally reduced the microaggregates induced by shear stress alone . In conclusion , microaggregates of platelets via Q9H244 receptors could play a key role in the hypersensitivity of platelets in diabetic patients , and the measurement of microaggregation could be a useful marker to estimate of thrombogenesis . These findings present a possible new means for patients with diabetes to prevent ischemic events . P23458 / P40763 activation directly inhibits IL-12 production in dendritic cells by preventing P50750 /P-TEFb recruitment to the p35 promoter . Inhibition of Janus-activated kinase-1 ( P23458 ) is a promising clinical concept for post-transplant immunosuppression and autoimmunity . However , it also raises concerns regarding possible immunosuppressive side effects . Our study investigates P23458 signalling in the context of P29965 and bacterially activated human MoDC using siRNA and biological inhibitors . We demonstrate that strong stimuli ( e.g. intact Escherichia coli or LPS in addition to IL-1β ) induce IL-12p70 via a ROS/ Q04206 / P50750 pathway that is inhibited by simultaneous P23458 / P40763 signalling . Transcription is effective if Q04206 recruits the positive transcription elongation factor b ( P-TEFb ) component P50750 to a combined Q04206 / P40763 binding site -50 to -20bp upstream of the start site of the IL-12p35 promoter . P40763 simultaneously attaches to this site and inhibits P50750 binding . In the presence of IFNγ , P23458 /2 inhibitors block P42224 / P10914 / Q02556 -dependent activation and simultaneously enhance P50750 -dependent activation signals . This inverse regulation of IFNγ- vs. E. coli-induced cytokine production by JAK inhibitors including DB08877 was similarly observed for P05231 and P01375 -α production , but not for P22301 production . Thus , P23458 inhibition enhances IL-12p70 production in this context by increased DNA binding of P50750 . In contrast , weak Q04206 -activation signals ( P29965 , LPS ) depended on IFN-γ induced P42224 / P10914 / Q02556 co-signalling , which was completely blocked by JAK inhibitors as reported before . Our results suggest a novel molecular mechanism of how cytokine responses to invading pathogens are separable from IFNγ-dependent autoimmunity by targeting P23458 / P40763 activation . Thirteen type I loci from HSA4q , HSA6p , HSA7q and HSA12q were comparatively Q5TCZ1 -mapped in four river buffalo and sheep chromosomes . Thirteen goat BAC clones containing coding sequences from HSA7 , HSA12q , HSA4 and HSA6p were fluorescence in situ mapped to river buffalo ( Bubalus bubalis , BBU ) and sheep ( Ovis aries , OAR ) R-banded chromosomes . The following type I loci were mapped : P03999 to BBU8q32 and OAR4q32 , P35523 to BBU8q34 and OAR4q34 , P17936 to BBU8q24 and OAR4q27 , KRT to BBU4q21 and OAR 3q21 , P01579 to BBU4q23 and OAR3q23 , IGF1 to BBU4q31 and OAR3q31 , P30968 to BBU7q32 and OAR6q32 , P55157 to BBU7q21 and OAR6q15 , P35913 to BBU7q36 and OAR6q36 , BF to BBU2p22 and OAR20q22 , P05305 to BBU2p24 and OAR20q24 , P08263 to BBU2p22 and OAR20q22 , OLADRB ( MHC ) to BBU2p22 and OAR20q22 . All mapped loci appeared to be located on homologous chromosomes and chromosome bands in both bovids . Comparison between gene orders in bovid ( BBU and OAR ) and human ( HSA ) chromosomes revealed complex rearrangements , especially between BBU7/OAR6 and HSA4 , as well as between BBU2p/OAR20 and HSA6p . Tunicate gonadotropin-releasing hormone ( DB00644 ) peptides selectively activate Ciona intestinalis DB00644 receptors and the green monkey type II P30968 . In vertebrates , DB00644 binds to its receptor and stimulates predominantly G(q/11)-mediated signal transduction in gonadotropes . However , little is known about the P30968 and its signaling pathway in tunicates , a group that arose before the vertebrates . Although tunicates have had duplications of a few genes in the last 600 million years , the early vertebrates had duplications of the full genome . Also unknown is the nature of DB00644 signaling in the tunicate , which lacks both a pituitary gland and sex steroids . However , we know that tunicates have DB00644 peptides because we previously reported six DB00644 peptides encoded within the tunicate genome of Ciona intestinalis . Here we clone and sequence cDNAs for four putative DB00644 receptors from C. intestinalis . These are the only invertebrate DB00644 receptors found to date . Each Ciona P30968 was expressed in COS-7 cells , incubated with each of the six C. intestinalis GnRHs and assayed for a signaling response . DB00644 receptors 1 , 2 , and 3 responded to Ciona DB00644 peptides to stimulate intracellular DB02527 accumulation . In contrast , only P30968 1 activated inositol phosphate turnover in response to one of the Ciona GnRHs . The green monkey type II P30968 cDNA was tested as a comparison and a positive control . In conclusion , the four DB00644 receptors encoded within the C. intestinalis genome were all transcribed into messenger RNA , but only three of the Ciona DB00644 receptors were biologically active in our assays . The Ciona DB00644 receptors almost exclusively activated the DB02527 pathway . 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 . 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 . Effects of metalloproteinase inhibition in a murine model of renal ischemia-reperfusion injury . Ischemia-reperfusion injury ( IRI ) is a leading cause of acute tubular necrosis ( ATN ) and delayed graft function in transplanted organs . Up-regulation of matrix metalloproteinases ( MMPs ) propagates the microinflammatory response that drives IRI . This study sought to determine the specific effects of Marimastat ( Vernalis , DB00786 ) , a broad spectrum MMP and P78536 inhibitor , on IRI-induced ATN . Mice were pretreated with Marimastat or methylcellulose vehicle for 4 d before surgery . Renal pedicles were bilaterally occluded for 30 min and allowed to reperfuse for 24 h . Baseline creatinine levels were consistent between experimental groups ; however , post-IRI creatinine levels were 4-fold higher in control mice ( p < 0.0001 ) . The mean difference between the post-IRI histology grades of Marimastat-treated and control kidneys was 1.57 ( p = 0.003 ) , demonstrating more severe damage to control kidneys . Post-IRI mean ( +/-SEM ) P08253 activity rose from baseline levels in control mice ( 3.62 +/- 0.99 ) ; however , pretreated mice presented only a slight increase in mean P08253 activity ( 1.57 +/- 0.72 ) ( p < 0.001 ) . In conclusion , these data demonstrate that MMP inhibition is associated with a reduction of IRI in a murine model . Interferon-gamma-induced dephosphorylation of P40763 and apoptosis are dependent on the P42345 pathway . Interferon-gamma ( P01579 ) exhibits diverse biological activities , including control of cell growth and tumor suppression . Here , we report that the treatment of M12 cells , a human metastatic prostate cancer cell line , with P01579 , resulted in marked inhibition of cell proliferation and induced apoptosis . These effects were not seen with either IFN-alpha or IFN-beta . M12 cells , like many other human cancer cells , contain constitutively activated signal transducer and activator of transcription 3 ( P40763 ) . The basal levels of both Akt and P27361 /2 phosphorylation are also markedly elevated in M12 cells . Strikingly , P01579 -induced apoptosis and growth inhibition of M12 cells were associated with persistent suppression of the constitutive tyrosine-phosphorylated P40763 ( pY- P40763 ) . The P01579 -induced dephosphorylation of pY- P40763 , however , was inhibited when the P42345 pathway was specifically blocked by rapamycin . Inhibition of PI-3K with low-dose LY294002 , or MAPK with PD98059 also suppressed the P42345 / P08133 S6k pathway , and correlated with the blockage of P01579 -induced dephosphorylation of pY- P40763 . Simultaneously , treatment with LY294002 , PD98059 , or rapamycin abolished P01579 -induced apoptosis in M12 cells . The inhibition of the P42345 pathway , however , did not affect P01579 -induced activation of P42224 pathway , and suppression of P42224 expression by siRNA had no effect on P01579 -induced dephosphorylation of pY- P40763 . Taken together , these results demonstrate that an intact P42345 pathway is critical for P01579 -induced suppression of pY- P40763 and apoptosis . Our study thus provides novel insights into the contributions of signaling pathways other than the classical JAK/ P42224 pathway in the anti-proliferative , proapoptotic actions of P01579 . Transcription inhibition by flavopiridol : mechanism of chronic lymphocytic leukemia cell death . DB03496 is active against chronic lymphocytic leukemia ( CLL ) cells in vitro and in the treatment of advanced stage disease , but the mechanisms of these actions remain unclear . Originally developed as a general cyclin-dependent kinase inhibitor , flavopiridol is a potent transcriptional suppressor through the inhibition of positive transcription elongation factor b ( P-TEFb ; P50750 /cyclin T ) . P-TEFb phosphorylates the C-terminal domain ( CTD ) of RNA polymerase II to promote transcriptional elongation . Because most CLL cells are not actively cycling , and their viability is dependent upon the continuous expression of antiapoptotic proteins , we hypothesized that flavopiridol induces apoptosis in CLL cells through the transcriptional down-regulation of such proteins . This study demonstrated that flavopiridol inhibited the phosphorylation of the CTD of RNA polymerase II in primary CLL cells and reduced RNA synthesis . This was associated with a decline of the transcripts and the levels of short-lived antiapoptotic proteins such as myeloid cell leukemia 1 ( Mcl-1 ) , and resulted in the induction of apoptosis . The B-cell lymphoma 2 ( Bcl-2 ) protein level remained stable , although its mRNA was consistently reduced , suggesting that the outcome of transcriptional inhibition by flavopiridol is governed by the intrinsic stability of the individual transcripts and proteins . The dependence of CLL-cell survival on short-lived oncoproteins may provide the biochemical basis for the therapeutic index in response to flavopiridol . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . 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 . 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 . Stimulation of purinergic receptors modulates chemokine expression in human keratinocytes . DB00171 is abundantly released from stressed or damaged cells in response to mechanical stimulation , bacteria , or noxious agents . In this study , we have investigated the possible involvement of P2 receptors ( receptor for extracellular nucleotides ) in the expression and release of inflammatory mediators by human keratinocytes . Notably , extracellular DB00171 displayed a complex regulation of P01579 -stimulated chemokine expression , with upregulation of CC chemokine ligand 2 ( P13500 ) , P13501 and CXC chemokine ligand 8 ( P10145 ) , and suppression of the receptor CXC chemokine receptor 3 ( P49682 ) , Q07325 , P02778 , and O14625 . The effect of DB00171 was mimicked by ADP and adenosine-5'-O-3-thiotriphosphate , whereas 2',3'-O-(4-benzoylbenzoyl) DB00171 ( BzATP ) downmodulated all chemokines investigated . UTP had no effect on P01579 -stimulated chemokine secretion . The broad-spectrum P2 receptor antagonist suramin and the selective P47900 inhibitor adenosine 3'-phosphate 5'-phosphosulfate counteracted the effect of DB00171 on secretion of all the chemokines examined , whereas pyridoxal phosphate 6-azophenyl 2',4'-disulfonic acid and KN62 ( 1- [ N,O-bis ( 5-isoquinoline sulfonyl ) -N-methyl-L-tyrosyl ] 4 phenylpiperazine ) partially prevented the inhibitory effect of DB00171 on P02778 secretion , but on the other hand potentiated the DB00171 -stimulatory effect on P13501 , P13500 , and P10145 release . In lesional skin of psoriasis and atopic dermatitis patients , intense Q99572 reactivity was confined to the cell membrane of the basal layer , whereas diffuse P47900 immunostaining was found throughout the epidermis . Collectively , our data suggest that the orchestrated activation of distinct P2Y and P2X receptors modulates skin inflammation . 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 . 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 . Interferon-induced Q13546 /RIP3-mediated necrosis requires P19525 and is licensed by Q13158 and caspases . Interferons ( IFNs ) are cytokines with powerful immunomodulatory and antiviral properties , but less is known about how they induce cell death . Here , we show that both type I ( α/β ) and type II ( γ ) IFNs induce precipitous receptor-interacting protein (RIP)1/RIP3 kinase-mediated necrosis when the adaptor protein Fas-associated death domain ( Q13158 ) is lost or disabled by phosphorylation , or when caspases ( e.g. , caspase 8 ) are inactivated . IFN-induced necrosis proceeds via progressive assembly of a Q13546 -RIP3 " necrosome " complex that requires Jak1/ P42224 -dependent transcription , but does not need the kinase activity of Q13546 . Instead , IFNs transcriptionally activate the RNA-responsive protein kinase P19525 , which then interacts with Q13546 to initiate necrosome formation and trigger necrosis . Although IFNs are powerful activators of necrosis when Q13158 is absent , these cytokines are likely not the dominant inducers of RIP kinase-driven embryonic lethality in Q13158 -deficient mice . We also identify phosphorylation on serine 191 as a mechanism that disables Q13158 and collaborates with caspase inactivation to allow IFN-activated necrosis . Collectively , these findings outline a mechanism of IFN-induced RIP kinase-dependent necrotic cell death and identify Q13158 and caspases as negative regulators of this process . DB01296 improves cardiac function following trauma-hemorrhage by increased protein O-GlcNAcylation and attenuation of NF-{kappa}B signaling . We have previously demonstrated that in a rat model of trauma-hemorrhage ( T-H ) , glucosamine administration during resuscitation improved cardiac function , reduced circulating levels of inflammatory cytokines , and increased tissue levels of O-linked N-acetylglucosamine ( O-GlcNAc ) on proteins . The mechanism(s) by which glucosamine mediated its protective effect were not determined ; therefore , the goal of this study was to test the hypothesis that glucosamine treatment attenuated the activation of the nuclear factor-kappaB ( NF-kappaB ) signaling pathway in the heart via an increase in protein O-GlcNAc levels . Fasted male rats were subjected to T-H by bleeding to a mean arterial blood pressure of 40 mmHg for 90 min followed by resuscitation . DB01296 treatment during resuscitation significantly attenuated the T-H-induced increase in cardiac levels of P01375 and P05231 mRNA , IkappaB-alpha phosphorylation , NF-kappaB , NF-kappaB DNA binding activity , P05362 , and P05164 activity . LPS ( 2 microg/ml ) increased the levels of IkappaB-alpha phosphorylation , P01375 , P05362 , and NF-kappaB in primary cultured cardiomyocytes , which was significantly attenuated by glucosamine treatment and overexpression of O-GlcNAc transferase ; both interventions also significantly increased O-GlcNAc levels . In contrast , the transfection of neonatal rat ventricular myocytes with O15294 small-interfering RNA decreased O-GlcNAc transferase and O-GlcNAc levels and enhanced the LPS-induced increase in IkappaB-alpha phosphorylation . DB01296 treatment of macrophage cell line RAW 264.7 also increased O-GlcNAc levels and attenuated the LPS-induced activation of NF-kappaB . These results demonstrate that the modulation of O-GlcNAc levels alters the response of cardiomyocytes to the activation of the NF-kappaB pathway , which may contribute to the glucosamine-mediated improvement in cardiac function following hemorrhagic shock .

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Evaluating the ligand specificity of zebrafish parathyroid hormone ( PTH ) receptors : comparison of PTH , PTH-related protein , and tuberoinfundibular peptide of 39 residues . Homologs of mammalian PTH1 and PTH2 receptors , and a novel PTH3 receptor have been identified in zebrafish ( zPTH1 , zPTH2 , and zPTH3 ) . zPTH1 receptor ligand specificity is similar to that of mammalian PTH1 receptors . The zPTH2 receptor is selective for PTH over PTH-related protein ( P12272 ) ; however , PTH produces only modest DB02527 accumulation . A P49190 -selective peptide , tuberoinfundibular peptide of 39 residues ( Q96A98 ) , has recently been purified from bovine hypothalamus . The effect of Q96A98 has not previously been examined on zebrafish receptors . The zPTH3 receptor was initially described as P12272 selective based on comparison with the effects of human PTH . We have now examined the ligand specificity of the zebrafish PTH-recognizing receptors expressed in COS-7 cells using a wide range of ligands . Q96A98 is a potent agonist for stimulation of DB02527 accumulation at two putative splice variants of the zPTH2 receptor ( EC50 , 2.6 and 5.2 nM ) ; in comparison , PTH is a partial agonist [ maximal effect ( Emax ) of PTH peptides ranges from 28-49 % of the Q96A98 Emax ] . As Q96A98 is much more efficacious than any known PTH-like peptide , a homolog of Q96A98 may be the zPTH2 receptor 's endogenous ligand . At the zPTH3 receptor , rat PTH-(1-34) and rat PTH-(1-84) ( EC50 , 0.22 and 0.45 nM ) are more potent than P12272 ( EC50 , 1.5 nM ) , and DB05829 -(1-34) binds with high affinity ( 3.2 nM ) . PTH has not been isolated from fish . P12272 -like peptides , which have been identified in fish , may be the natural ligands for zPTH1 and zPTH3 receptors . 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 . A role for parathyroid hormone-related protein in the pathogenesis of inflammatory/autoimmune diseases . Our increased understanding of the critical role of cytokines in chronic inflammatory/autoimmune diseases has led to the recent development of effective anti-cytokine treatments . In particular , agents blocking the function of P01375 , a cytokine first identified as an endotoxin-inducible mediator of tumor cell necrosis , are now licensed for the treatment of rheumatoid arthritis ( RA ) and inflammatory bowel disease . However , P01375 is but one member of a cytokine network that is responsible for mediating these inflammatory disorders . Therefore , as our understanding of the pathophysiologic role of other members of this inflammatory network increases , other cytokines may similarly be identified as effective targets for treatment . In this article , we will review evidence which suggests that parathyroid hormone-related protein ( P12272 ) , a peptide which , like P01375 , was first identified because of its effects in the setting of malignancy , may in fact serve an important non-neoplastic , physiologic function by mediating the inflammatory/autoimmune host response . Data identifying P12272 as a member of the cytokine network induced in multi-organ inflammation and rheumatoid arthritis will be summarized , initial evidence comparing the therapeutic efficacy of P12272 - vs. P01375 -blockade in the treatment of endotoxemia will be reviewed , and potential future areas of research , including assessment of the effects of P12272 blockade in the treatment of RA , will be discussed . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . MR properties of excised neural tissue following experimentally induced inflammation . Changes in the MR parameters of inflamed neural tissue were measured in vitro . P01375 -alpha ( P01375 ) was injected into rat sciatic nerves to induce inflammation with negligible axonal loss and demyelination . The MR parameters , such as T1/ P24752 relaxation and magnetization transfer ( MT ) , were measured 2 days after P01375 injection and were found to be substantially different from those of normal nerves . The average T1/ P24752 relaxation times increased , whereas the MT ratio ( Q99707 ) and the quantitative MT parameter M0B ( which describes the semisolid pool of protons ) decreased . The MR parameters correlated very well with the extracellular volume fraction ( EM ) of neural tissue evaluated by quantitative histopathology . The multicomponent P24752 relaxation was shown to provide the best quantitative assessment of changes in neural tissue microstructure , and allowed us to distinguish between the processes of inflammation and demyelination . In comparison , the MT measurements were less successful due to competing contributions of demyelination and pH-sensitive changes in the MT effect . Mechanism of action of transmembrane activator and calcium modulator ligand interactor-Ig in murine systemic lupus erythematosus . B cell-activating factor belonging to the P01375 family ( Q9Y275 ) blockade prevents the onset of disease in systemic lupus erythematosus ( SLE ) -prone NZB/NZW F(1) mice . To determine the mechanism of this effect , we administered a short course of O14836 -Ig with and without six doses of P16410 -Ig to 18- to 20-wk-old NZB/NZW F(1) mice and evaluated the effect on B and T cell subsets and on anti-dsDNA Ab-producing B cells . Even a brief exposure to O14836 -Ig had a beneficial effect on murine SLE ; P16410 -Ig potentiated this effect . The combination of O14836 -Ig and P16410 -Ig resulted in a temporary decrease in serum IgG levels . However , after cessation of treatment , high titers of IgG anti-dsDNA Abs appeared in the serum and IgG Abs deposited in the kidneys . Despite the appearance of pathogenic autoantibodies , the onset of proteinuria was markedly delayed ; this was associated with prolonged depletion of B cells past the T1 stage , a decrease in the size of the spleen and lymph nodes , and a decrease in the absolute number of activated and memory P01730 (+) T cells . O14836 -Ig treatment normalized serum levels of IgM that are markedly elevated in NZB/W F(1) mice ; this appeared to be due to a prolonged effect on the ability of the splenic microenvironment to support short-lived IgM plasma cells . Finally , a short course of combination O14836 -Ig and P16410 -Ig prolonged life and even reversed proteinuria in aged NZB/W F(1) mice , suggesting that Q9Y275 blockade may be an effective therapeutic strategy for active SLE . JNK regulatory molecule Q969Q6 induces IgG autoantibody-producing plasmablasts from peritoneal B1a cells . Peritoneal B1a cells expressing P06127 and CD11b generate autoantibody-producing precursors in autoimmune-prone mice . Previous studies show reduced JNK signaling in peritoneal B1a cells of female New Zealand Black mice and an abnormal increase of protein phosphatase 2A subunit Q969Q6 that regulates P11274 -mediated JNK signaling as a cause of autoimmunity . To investigate the mechanism regulating B1a differentiation into autoantibody-secreting plasmablasts ( PBs ) , we applied an in vitro culture system that supports long-term growth of germinal center ( GC ) B cells ( iGB ) with P05112 , P29965 , and Q9Y275 . Compared with spleen B2 cells , B1a cells differentiated into GC-like B cells , but more markedly into PBs , and underwent class switching toward IgG1 . During iGB culture , B1a cells expressed GC-associated aicda , g5pr , and bcl6 , and markedly PB-associated prdm1 , irf4 , and xbp1 . B1a-derived iGB cells from New Zealand Black × New Zealand White F1 mice highly differentiated into autoantibody-secreting PBs in vitro and localized to the GC area in vivo . In iGB culture , JNK inhibitor SP600125 augmented the differentiation of C57BL/6 B1a cells into PBs . Furthermore , B1a cells from Q969Q6 transgenic mice markedly differentiated into IgM and IgG autoantibody-secreting PBs . In conclusion , JNK regulation is critical to suppress autoantibody-secreting PBs from peritoneal B1a cells . 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 . Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 receptor ( IL13Rα2 ) that differs from the physiological P24394 /IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL-13-PE ) encoding a mutated human P35225 fused to Pseudomonas exotoxin ( mhIL-13-PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 that binds to the physiological P24394 /IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL-13-PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL-13-PE and two protein formulations , hIL-13-PE used in clinical trials ( DB05305 ) and a second-generation mhIL-13-PE . DB05305 doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL-13-PE led to ∼40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL-13-PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 include its short half-life , which demanded frequent or continued administration , and binding to P24394 /IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM . The role of the polyol pathway in acute kidney injury caused by hindlimb ischaemia in mice . The polyol pathway , a collateral glycolytic process , previously considered to be active in high glucose milieu , has recently been proposed to play a crucial role in ischaemia/reperfusion tissue injury . In this study , we explored the role of the polyol pathway in acute kidney injury ( AKI ) , a life-threatening condition , caused by hindlimb ischaemia , and determined if inhibition of the polyol pathway by aldose reductase ( AR ) inhibitor is beneficial for this serious disorder . Mice 8 weeks of age rendered hindlimb ischaemic for 3 h by the clipping of major supporting arteries revealed marked muscle necrosis with accumulation of sorbitol and fructose in ischaemic muscles . Serum concentrations of blood urea nitrogen ( BUN ) , creatinine phosphokinase ( CPK ) , creatinine , tumour necrosis factor ( P01375 ) -alpha as well as interleukin ( IL ) -6 were all elevated in these mice . Treatment with AR inhibitor ( Q9Y4X5 ) effectively suppressed muscle necrosis and accompanying inflammatory reactions and prevented renal failure . Similar to Q9Y4X5 -treated mice , AR-deficient mice were protected from severe ischaemic limb injury and renal failure , showing only modest muscle necrosis and significant suppression of serum markers of renal failure and inflammation . Thus , these findings suggest that the polyol pathway is implicated in AKI caused by ischaemic limb injury and that AR may be a potential therapeutic target for this condition . P02751 supports P01375 -induced osteopontin expression through beta1 integrin and P29323 in HN-22 cells . The extracellular matrix ( Q13201 ) , in collaboration with intracellular signal , plays a critical role in the modulation of cellular behavior and function . Herein , we investigated the influence of fibronectin ( FN ) and tumor necrosis factor-alpha ( P01375 ) on P10451 expression in HN-22 , a human head and neck squamous cell carcinoma ( HNSCC ) cell line . The data showed that P01375 significantly increased P10451 expression only in the FN-coated condition . Application of function-blocking antibody directed against beta1 integrin abolished this P10451 induction . Moreover , P01375 when added together with activating beta1 integrin antibody is sufficient to induced P10451 expression . The combination effect of FN and P01375 was significantly deteriorated by a MEK inhibitor , but not NF-kappaB inhibitor . We further demonstrated that the phosphorylation of P27361 /2 was strongly enhanced by P01375 and FN compared to the application of either one alone . Synergistic effect on P27361 /2 phosphorylation was also detected by P01375 and activating beta1 integrin antibody , whereas inhibitory antibody to beta1 integrin attenuated FN and P01375 -induced phosphorylation of P27361 /2 . Our results indicate that FN coordinates P01375 -mediated P10451 induction via beta1 integrin-dependent signaling mechanism that activates P29323 . The results suggest the critical role of tumor micro-environment signaling networks on the regulation of cytokine expression profiles during tumor progression . Homocysteine Level and Mechanisms of Injury in Parkinson 's Disease as Related to P42898 , Q99707 , and P11586 Genes Polymorphisms and L-Dopa Treatment . An elevated concentration of total homocysteine ( tHcy ) in plasma and cerebrospinal fluid is considered to be a risk factor for Alzheimer 's disease ( AD ) and Parkinson 's disease ( PD ) . Homocysteine ( Hcy ) levels are influenced by folate concentrations and numerous genetic factors through the folate cycle , however , their role in the pathogenesis of PD remains controversial . Hcy exerts a neurotoxic action and may participate in the mechanisms of neurodegeneration , such as excitotoxicity , oxidative stress , calcium accumulation , and apoptosis . Elevated Hcy levels can lead to prooxidative activity , most probably through direct interaction with N-methyl-D-aspartate ( DB01221 ) receptors and sensitization of dopaminergic neurons to age-related dysfunction and death . Several studies have shown that higher concentration of Hcy in PD is related to long-term administration of levodopa ( L-dopa ) . An elevation of plasma tHcy levels can also reflect deficiencies of cofactors in remethylation of Hcy to methionine ( DB00134 ) ( folates and vitamin B12 ) and in its transsulfuration to cysteine ( DB00151 ) ( vitamin B6 ) . It is believed that the increase in the concentration of Hcy in PD can affect genetic polymorphisms of the folate metabolic pathway genes , such as P42898 ( C677T , A1298C and G1793A ) , Q99707 ( A2756G ) , and P11586 ( G1958A ) , whose frequencies tend to increase in PD patients , as well as the reduced concentration of B vitamins . In PD , increased levels of Hcy may lead to dementia , depression and progression of the disease . 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 . 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 . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . 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 . 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 . 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 . 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 . Selective activation of O14836 by syndecan-2 . B-lymphocyte homeostasis and function are regulated by complementary actions of the TNFR family members O14836 , Q02223 , and Q96RJ3 , which are expressed by mature B cells . How these receptors are differentially activated is not entirely understood , because the primary ligand Q9Y275 binds to all three . We searched for alternative ligands for O14836 using recombinant O14836 -Fc fusion protein as a probe and identified syndecan-2 as a new binding partner . O14836 binding appears to require heparan sulfate posttranslational modifications of syndecan-2 , because free heparin or pretreatment with heparitinase blocked the interaction . P34741 bound O14836 but bound neither Q96RJ3 nor Q02223 . Transfected cells expressing syndecan-2 activated signaling through O14836 , as indicated by an NFAT-specific reporter . P18827 and syndecan-4 were also able to induce O14836 signaling in a similar manner . This is the first identification of ligands that selectively activate O14836 without simultaneously triggering Q02223 or Q96RJ3 . This finding may help explain the alternative outcomes of signaling from this family of receptors in B cells .

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Multiplex imaging and cellular target identification of kinase inhibitors via an affinity-based proteome profiling approach . DB05220 is a highly potent and presumably selective inhibitor of O14965 ( AKA ) and has shown promising antitumor activities . Like other kinase inhibitors which target the DB00171 -binding site of kinases , DB05220 might be expected to have potential cellular off-targets . Herein , we report the first photoaffinity-based , small molecule AKA probe capable of both live-cell imaging of AKA activities and in situ proteome profiling of potential off-targets of DB05220 ( including AKA-associating proteins ) . By using two mutually compatible , bioorthogonal reactions ( copper-catalyzed azide-alkyne cycloaddition chemistry and TCO-tetrazine ligation ) , we demostrate small molecule-based multiplex bioimaging for simultaneous in situ monitoring of two important cell-cycle regulating kinases ( AKA and P06493 ) . A broad range of proteins , as potential off-targets of DB05220 and AKA's-interacting partners , is subsequently identified by affinity-based proteome profiling coupled with large-scale LC-MS/MS analysis . From these studies , we discover novel AKA interactions which were further validated by cell-based immunoprecipitation ( IP ) experiments . The effect of alefacept on T-cell subsets and cells expressing NK receptors in lesional psoriatic skin : the effects of monotherapy and combination treatment with calcipotriol . OBJECTIVES : To investigate the effect of weekly alefacept monotherapy 15 mg i.m. on epidermal hyperproliferation , keratinization , T-cell subsets and cells expressing NK receptors during 12 weeks of treatment . Furthermore , the addition of calcipotriol cream to alefacept treatment was studied and compared with alefacept monotherapy . METHODS : Five patients participated in this study , and used weekly alefacept 15 mg i.m. combined with calcipotriol cream up to a maximum of 100 g per week . Biopsies from two lesions ( one treated and another lesion not treated with calcipotriol cream ) were taken at week 0 and week 12 . We investigated the number of T-cell subsets ( P01730 , CD8 , CD45RO , CD45RA , P06729 , CD25 ) , cells expressing NK receptors ( Q13241 and CD161 ) , the proliferation marker Ki-67 and the keratinization marker keratin-10 . RESULTS : DB00092 monotherapy induced a statistically significant reduction of P01730 + , CD45RO+ and P06729 + cells in dermis and epidermis and CD8+ cells in epidermis at week 12 . Furthermore , the keratin-10 positive epidermal surface was significantly increased at week 12 . The combination of alefacept and calcipotriol cream induced a statistically significant reduction of only P01730 + and CD45RO+ cells at week 12 . CONCLUSIONS : The number of memory effector T-cells in the psoriatic lesion is significantly decreased by alefacept , and calcipotriol cream does not seem to have an additional effect on this reduction . Cells expressing NK receptors are virtually not targeted by alefacept monotherapy or the combination . 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 . A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors . P10721 or alpha-platelet-derived growth factor receptor ( alpha- P09619 ) activating mutations are the pathogenic mechanisms that characterize gastrointestinal stromal tumors ( GIST ) . Despite excellent responses to imatinib mesylate ( IM ) , patients are relapsing . We developed an IM-resistant GIST cell line ( GIST-R ) from the IM-sensitive GIST882 cell line ( GIST-S ) by growing these cells in IM . Gene expression profiling ( GEP ) of GIST-S , GIST-R cells and two IM resistant GIST patients demonstrated that P10721 is downregulated implying a major role in IM resistance . Instead , GIST-R cells have acquired IM resistance by overexpressing the oncogenic receptor tyrosine kinase - P30530 - in a ' kinase switch ' . Further , the two IM resistant GIST patients express P30530 and not c-Kit , seen by immunohistochemistry ( IHC ) . Real time reverse transcriptase-polymerase chain reaction and Western blotting of the GIST-S and GIST-R cells confirmed the switch from Kit to P30530 . In GIST-R , P30530 is tyrosine phosphorylated and its ligand growth-arrest-specific gene 6 is overexpressed implying autocrine activation . The kinase switch is associated with a morphological change from spindle to epithelioid . Molecular modeling of the kinase domain of mutant c-Kit ( V654A ) and P30530 showed no binding to IM but efficient binding to DB05216 , a novel c-Kit/ P30530 kinase inhibitor . DB05216 synergizes with docetaxel ( taxotere ) and is cytotoxic to GIST cells . 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 . 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 . DB09073 ( PD 0332991 ) : targeting the cell cycle machinery in breast cancer . INTRODUCTION : The cyclin D-cyclin-dependent kinases 4 and 6 ( P11802 /6 ) -retinoblastoma ( P06400 ) pathway , governing the cell cycle restriction point , is frequently altered in breast cancer and is a potentially relevant target for anticancer therapy . DB09073 ( PD 0332991 ) , a potent and selective inhibitor of P11802 and Q00534 , inhibits proliferation of several P06400 -positive cancer cell lines and xenograft models . AREAS COVERED : The basic features and abnormalities of the cell cycle in breast cancer are described , along with their involvement in estrogen signaling and endocrine resistance . The pharmacological features of palbociclib , its activity in preclinical models of breast cancer and the potential determinants of response are then illustrated , and its clinical development in breast cancer described . A literature search on the topic was conducted through PubMed and the proceedings of the main cancer congresses of recent years . EXPERT OPINION : The combination of palbociclib with endocrine agents is a very promising treatment and Phase III clinical trials are ongoing to confirm its efficacy . Further , potentially useful combinations are those with drugs targeting mitogenic signaling pathways , such as P04626 - and PI3K-inhibitors . Combination with chemotherapy seems more problematic , as antagonism has been reported in preclinical models . The identification of predictive factors , already explored in preclinical studies , must be further refined and validated in clinical trials . Recombinant human prothrombin kringle-2 induces bovine capillary endothelial cell cycle arrest at G0- P55008 phase through inhibition of cyclin D1/ P11802 complex : modulation of reactive oxygen species generation and up-regulation of cyclin-dependent kinase inhibitors . P00734 is a plasma glycoprotein involved in blood coagulation and , as we have previously reported , prothrombin kringles inhibit BCE ( bovine capillary endothelial ) cell proliferation . To reveal the mechanism , we investigated the influence of rk-2 ( recombinant human prothrombin kringle-2 ) on the BCE cell cycle progression and ROS ( reactive oxygen species ) generation using FACS ( fluorescence-activated cell sorter ) analysis . Cell cycle analysis showed a decrease of G(1) phase cells in cells treated with P09038 ( basic fibroblast growth factor ) and an increase in cells treated with rk-2 , as compared with the control cells . But , the portion of the S phase was reversed . In Western blot analysis , P09038 induced cytoplasmic translocation of P38936 (Waf1/Cip1) and p27(Kip1) and phosphorylation of p27(Kip1) but rk-2 treatment inhibited translocation of P38936 (Waf1/Cip1) and p27(Kip1) from nucleus to cytoplasm and phosphorylation of p27(Kip1) . Also , rk-2 induced up-regulation of p53 and nuclear P38936 (Waf1/Cip1) and inhibited the cyclin D1/ P11802 ( cyclin-dependent kinase 4 ) complex . The ROS level of rk-2-treated BCE cells was increased 2-fold when compared with the control , but treatment with Q9C000 ( N-Acetyl-L : -cysteine ) , an anti-oxidant , decreased ROS generation about 55 % as compared with the rk-2 treatment . Q9C000 treatment also restored cell cycle progression inhibited by rk-2 and down-regulated p53 and nuclear P38936 (Waf1/Cip1) expression induced by rk-2.These data suggest that rk-2 induces the BCE cell cycle arrest at G(0)-G(1) phase through inhibition of the cyclin D1/ P11802 complex caused by increase of ROS generation and nuclear cyclin-dependent kinase inhibitors . 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 . DB05051 , a phytochemical extract from the Scutellaria barbata plant , disrupts proliferation of human breast and prostate cancer cells through distinct mechanisms dependent on the cancer cell phenotype . DB05051 is an aqueous extract from the Scutellaria barbata plant shown to have anticancer properties in a variety of human cancers . In order to determine its efficacy on human reproductive cancers , we assessed the responses of two human breast cancer cell lines , estrogen sensitive MCF7 and estrogen insensitive MDA-MB-231 , and of two human prostate cancer cell lines , androgen sensitive LNCaP and androgen insensitive PC3 which are human cell lines that represent early and late stage reproductive cancers . DB05051 inhibited reproductive cancer growth in all cell lines by regulating expression levels of key cell cycle components that differ with respect to the cancer cell phenotypes . In early stage estrogen sensitive MCF7 cells , DB05051 induced a G₁ cell cycle arrest and ablated expression of key G₁ cell cycle regulators P12004 D1 , P24941 and P11802 , as well as growth factor stimulatory pathways and estrogen receptor-α expression . Transfection of luciferase reporter plasmids revealed that the loss of P24941 , P11802 and estrogen receptor-α transcript expression resulted from the BZL-dependent ablation of promoter activities . DB05051 growth arrests early stage androgen sensitive LNCaP cells in the G₂/M phase with corresponding decreases in P12004 B1 , P06493 and androgen receptor expression . In late stage hormone insensitive breast ( MDA-MB-231 ) and prostate ( PC3 ) cancer cells , DB05051 induced an S phase arrest with corresponding ablations in P12004 A2 and P24941 expression . Our results demonstrate that DB05051 exerts phenotype specific anti-proliferative gene expression responses in human breast and prostate cancer cells , which will be valuable in the potential development of BZL-based therapeutic strategies for human reproductive cancers . DB00115 responsive homocystinuria and megaloblastic anemia : heterogeneity in methylcobalamin deficiency . A male infant with methyl-B12 deficiency ( cblE ) presented at age 6 weeks with lethargy , staring spells , and vomiting . He later became hypotonic and unresponsive to stimuli and required intubation and ventilation . He had homocystinuria and hypomethioninemia with megaloblastic anemia but normal serum folate and vitamin B12 concentrations . No methylmalonic aciduria was detected . Fibroblasts , cultured from the patient , were unable to grow in medium in which homocysteine replaced methionine and incorporated abnormally small amounts of [ 14C ] -methyl- DB00116 but normal amounts of [ 14C ] -propionate into protein . Methyl-B12 content of fibroblasts was low , while the adenosyl-B12 content was normal . Q99707 activity was decreased when the assay was performed under both optimal and suboptimal reducing conditions , suggesting heterogeneity in the cblE disease . The patient responded dramatically to hydroxocobalamin treatment . Homocystinuria disappeared after 10 days of therapy , and methionine was normalized after 3 weeks . Psychometric testing at age 15 months showed a developmental age of 9 months . [ Genetic risk factors of neural tube defects ] . Neural tube defects ( NTDs ) are a group of diseases caused by a failure of closure of the neural tube . Its aetiology contains both environmental and genetic factors . NTDs have a polygenic background . Genes , which are linked with NTDs occurrence , are both directly and indirectly connected with controlling the process of closure of the neural tube . Ones of those are genes of metabolism of folic acid as P42898 , Q99707 , Q9UBK8 , P35520 , P11586 , folic acid receptors ( FR ) regulator genes from PAX family , T , P16234 and P38398 genes . DB01174 -applied flagellin is internalized by polarized intestinal epithelial cells and elicits immune responses via the O60602 dependent mechanism . Bacteria release flagellin that elicits innate responses via O60602 ( O60602 ) . Here , we investigated the fate of apically administrated full length flagellin from virulent and avirulent bacteria , along with truncated recombinant flagellin proteins in intestinal epithelial cells and cellular responses . Flagellin was internalized by intestinal epithelial cell ( IEC ) monolayers of IEC-18 . Additionally , apically applied flagellin was internalized by polarized human Caco-2BBe and T-84 cells in a O60602 dependent mechanism . More , flagellin exposure did not affect the integrity of intestinal monolayers . With immunofluorescent staining , internalized flagellin was detected in both early endosomes as well as lysosomes . We found that apical exposure of polarized Caco-2BBe and T-84 to flagellin from purified Salmonella , Escherichia coli O83:H1 ( isolate from Crohn 's lesion ) or avirulent E. coli K12 induced comparable levels of basolateral P10145 secretion . A recombinant protein representing the conserved amino ( N ) and carboxyl ( C ) domains ( D ) of the flagellin protein ( P03886 /2ECHCD2/1 ) induced P10145 secretion from IEC similar to levels elicited by full-length flagellins . However , a recombinant flagellin protein containing only the D3 hypervariable region elicited no P10145 secretion in both cell lines compared to un-stimulated controls . Silencing or blocking O60602 in Caco-2BBe cells resulted in a lack of flagellin internalization and decreased P10145 secretion . Furthermore , apical exposure to flagellin stimulated transepithelial migration of neutrophils and dendritic cells . The novel findings in this study show that luminal-applied flagellin is internalized by normal IEC via O60602 and co-localizes to endosomal and lysosomal compartments where it is likely degraded as flagellin was not detected on the basolateral side of IEC cultures . Blood-brain barrier opened by stimulation of the parasympathetic sphenopalatine ganglion : a new method for macromolecule delivery to the brain . OBJECT : Drug delivery across the blood-brain barrier remains a significant challenge . Based on earlier findings , the authors hypothesized that parasympathetic innervation of the brain vasculature could be used to augment drug delivery to the brain . METHODS : Using a craniotomy-cerebrospinal fluid superfusate paradigm in rats with an intravenous injection of tracer the authors demonstrated that stimulation of the postganglionic parasympathetic fibers of the sphenopalatine ganglion ( Q9HBG6 ) increased the concentration of fluorescein isothiocyanate-dextran ( 4-250 kD ) in the superfusate by two- to sixfold . A histological examination indicated the presence of dextran in the parenchyma . In another experiment the amount of Evans blue dye in the brain following Q9HBG6 activation was similarly significantly elevated . The chemotherapeutic agents anti- P04626 monoclonal antibody and etoposide were also delivered to the brain and reached therapeutic concentrations . Brain homeostasis was not disturbed by this procedure ; a measurement of nicotinamide adenine dinucleotide reduction did not show a decrease in the tissue metabolic state and brain water content did not increase significantly . CONCLUSIONS : Sphenopalatine ganglion activation demonstrates a promising potential for clinical use in the delivery of small and large molecules to the brain . 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 . 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 . The zinc-finger protein Q9P0J7 is overexpressed during pancreatic cancer development and downregulation of Q9P0J7 inhibits pancreatic cancer development in mice . Q9P0J7 1 ( Q9P0J7 ) was found upregulated in a differential screen in the metaplastic epithelium in the pancreas of transforming growth factor ( TGF ) -alpha transgenic mice . Expression analysis indicated broad overexpression in human cancer tissues . Therefore , we investigated the hypothesis that Q9P0J7 promotes metaplastic changes and tumor development . Q9P0J7 represents an evolutionarily highly conserved protein with a 95 % identity between human and zebrafish . Q9P0J7 is expressed during embryonic development and in the majority of adult tissues investigated . Upregulation of nuclear Q9P0J7 expression is evident in preneoplastic lesions and in several epithelial malignancies , such as pancreatic cancer in mice and humans . In cell culture and in the chicken chorioallantoic membrane model , Q9P0J7 enhances proliferation , migration and invasion of P29320 -293 and Panc1 cells . In crossbreeding experiments , Q9P0J7 -knockdown gene trap mice showed a reduced number and size of premalignant lesions and absence of pancreatic cancer formation in TGF-alpha transgenic mice . This effect is related to the decreased expression of P55008 to S cell-cycle regulators such as cyclin D and cyclin-dependent kinase ( CDK ) 4 . Our data support the hypothesis that Q9P0J7 mediates pro-oncogenic functions in vitro and in vivo and downregulation of Q9P0J7 results in the inhibition of pancreatic cancer formation in mice . These effects are mediated through downregulation of cell-cycle control genes such as cyclin D and P11802 . 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 . Rapid mtDNA deletion by oxidants in rat liver mitochondria after hemin exposure . The amounts of superoxide and hydrogen peroxide generated by mitochondria under physiological conditions can be enhanced by cellular stress . This study tested the hypothesis that the response to hemin-induced stress , which includes heme oxygenase-1 ( P09601 ) induction , predisposes to oxidative damage of mitochondrial DNA ( mtDNA ) . Hepatic mitochondria from control , hemin- , and CO-exposed rats were incubated with tert-butyl hydroperoxide ( tert-BH ) or the NO donor 1,2,3,4-oxatriazolium , 5-amino-3- (3,4-dichlorophenyl)-chloride ( GEA 3162 ) . Mitochondrial total and oxidized glutathione ( DB00143 and GSSG ) , total and free iron , and 8-oxo-7 , 8-dihydro-2 ' deoxyguanosine ( 8-OHdG ) were determined with and without oxidants . As expected , oxidation by tert-BH induced significant DB00143 depletion and increased amounts of free iron and 8-OhdG . Oxidant exposure rapidly produced a large mtDNA deletion involving the coding regions for cytochrome c oxidase ( P36551 1 ) and DB00157 dehydrogenase ( P03886 and P03891 ) . DB03404 and CO greatly exacerbated susceptibility to the deletion of mtDNA by tert-BH , and this was attenuated by preincubation with DB00143 methyl ester . Analysis of mitochondria-associated proteins Bax and Bcl-xl in hemin- and CO-exposed rats showed significant responses , revealing interactions with apoptotic pathways . Thus , hemin-induced mitochondrial events sensitize a specific region of the mitochondrial genome to deletion , which is related to depletion of DB00143 and is not explained by effects of CO . This mtDNA damage is associated with altered expression of mitochondrial cell death proteins , thereby suggesting a novel mechanism for systemic or environmental pro-oxidants to influence apoptosis . 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 . Immunoregulatory and anti-tumor effects of polysaccharopeptide and Astragalus polysaccharides on tumor-bearing mice . The aim of this study was to determine whether polysaccharopeptide ( PSP ) and Astragalus polysaccharides ( APS ) can be combined together as a new complex prescription ( PSP + APS ) for aiding adriamycin ( AMD ) chemotherapy . Ehrlich 's ascites carcinoma ( EAC ) was used to establish a solid tumor model in Kunming mice . Immunocytochemical and immunohistochemical analysis were employed to detect the immunoregulatory and anti-tumor effects of EAC bearing mice after 30 days of administration with PSP and APS . PSP and PSP + APS could significantly increase the percentage of CD3(+) and P01730 (+) T-lymphocytes , the ratio of P01730 (+)/CD8(+) , and the expression of P60568 /IL-2R in spleen and Bax in tumor tissue , but led to a diminution of Bcl-2 and P11802 in tumor tissue compared with those of control group . In addition , PSP +APS could restore the immunological effects against AMD-induced immunosuppression , such as the subset of leukomonocyte , the expression of P60568 /IL-2R in the spleen , and the thymus index . These findings suggest that the immunomodulatory and anti-cancer effects of this new formula ( PSP+APS ) were better than those of PSP alone , and also could resist immunosuppression induced by AMD .

MH_train_1410

MH_train_1410

MH_train_1410

interacts_with DB00461?

multiple_choice

Activation of cellular invasion by trefoil peptides and src is mediated by cyclooxygenase- and thromboxane A2 receptor-dependent signaling pathways . We have investigated the possible functional relationships between cellular invasion pathways induced by trefoil factors ( TFFs ) , src , and the cyclooxygenases P23219 and P35354 . Pharmacological inhibitors of the Rho small GTPase ( P01024 exoenzyme ) , phospholipase C ( U-73122 ) , cyclooxygenases ( SC-560 , NS-398 ) , and the thromboxane A2 receptor ( P21731 ) antagonist SQ-295 completely abolished invasion induced by intestinal trefoil factor , pS2 , and src in kidney and colonic epithelial cells MDCKts.src and PCmsrc . In contrast , invasion was induced by the P21731 mimetic U-46619 , constitutively activated forms of the heterotrimeric G-proteins Galphaq ( AGalphaq ) , Galpha12 , Galpha13 ( AGalpha12/13 ) , which are signaling elements downstream of P21731 . Ectopic overexpression of pS2 cDNA and protein in MDCKts.src-pS2 cells and human colorectal cancer cells HCT8/S11-pS2 initiate distinct invasion signals that are Rho independent and P36551 and P21731 dependent . We detected a marked induction of P35354 protein and accumulation of the stable PGH2/TXA2 metabolite TXB2 in the conditioned medium from cells transformed by src . This led to activation of the P21731 -dependent invasion pathway , which is monitored via a Rho- and Galpha12/Galpha13-independent mechanism using the Galphaq/PKC signaling cascade . These findings identify a new intracrine/paracrine loop that can be monitored by TFFs and src in inflammatory diseases and progression of colorectal cancers . 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 . Analgesic and Anti-Inflammatory Activities of Methanol Extract of Cissus repens in Mice . The aim of this study was to investigate possible analgesic and anti-inflammatory mechanisms of the CR(MeOH) . Analgesic effect was evaluated in two models including acetic acid-induced writhing response and formalin-induced paw licking . The anti-inflammatory effect was evaluated by λ-carrageenan-induced mouse paw edema and histopathologic analyses . The results showed that CR(MeOH) ( 500 mg/kg ) decreased writhing response in the acetic acid assay and licking time in the formalin test . CR(MeOH) ( 100 and 500 mg/kg ) significantly decreased edema paw volume at 4th to 5th hours after λ-carrageenan had been injected . Histopathologically , CR(MeOH) abated the level of tissue destruction and swelling of the edema paws . These results were indicated that anti-inflammatory mechanism of CR(MeOH) may be due to declined levels of NO and MDA in the edema paw through increasing the activities of SOD , GPx , and GRd in the liver . Additionally , CR(MeOH) also decreased IL-1β , P05231 , NFκB , P01375 -α , P35354 , and P35228 levels . The contents of two active ingredients , ursolic acid and lupeol , were quantitatively determined . This paper demonstrated possible mechanisms for the analgesic and anti-inflammatory effects of CR(MeOH) and provided evidence for the classical treatment of Cissus repens in inflammatory diseases . Epigenetics , an early event in the modulation of gene expression by inositol hexaphosphate in ethylnitrosourea exposed mouse lungs . Mechanisms of anticancer effects of inositol hexaphosphate are not fully understood . Epigenetic changes are the early changes in tumorigenesis . DNA methyl transferases , methyl CpG binding proteins , methyl CpG DNA binding domain protein , and histone deacetylases are the major molecules involved in epigenetics . We have shown the effects of IP6 at the molecular level in mouse lungs before the tumor is developed . After 3 mo of ENU exposure , there was no tumor formation , but there was hyperplasia and lymphocytic infiltration in the lungs . Inflammation and DNA damage repair enzymes P35354 and P40692 appear to be upregulated , whereas tumor suppressor gene p16 was downregulated by ENU . On the other hand , ENU exposure more or less upregulated the epigenetic events such as the expressions of P26358 , MeCP2 , Q9UIS9 , and Q13547 . This alteration was reduced by IP6 administration . Results were supported by modulation of global DNA methylation and the modulation of promoter CpG methylation of p16 , P40692 , and P35354 genes . Hence , this study indicates the possible role of epigenetics at the early stage of tumor development and in the regulation of gene expression by IP6 before the onset of ENU-induced lung tumors . Biological differences between in vitro produced bovine embryos and parthenotes . Parthenotes may represent an alternate ethical source of stem cells , once biological differences between parthenotes and embryos can be understood . In this study , we analyzed development , trophectoderm ( TE ) differentiation , apoptosis/necrosis , and ploidy in parthenotes and in vitro produced bovine embryos . Subsequently , using real-time PCR , we analyzed the expression of genes expected to underlie the observed differences at the blastocyst stage . In vitro matured oocytes were either fertilized or activated with ionomycin +6-DMAP and cultured in simple medium . Parthenotes showed enhanced blastocyst development and diploidy and reduced TE cell counts . Apoptotic and necrotic indexes did not vary , but parthenotes evidenced a higher relative proportion of apoptotic cells between inner cell mass and TE . The pluripotence-related Q01860 and the methylation Q9Y6K1 genes were downregulated in parthenotes . Among pregnancy recognition genes , TP-1 was upregulated in parthenotes , while O00264 and PLAC8 did not change . Expression of p66(shc) and Q07812 / P10415 ratio were higher , and p53 lower , in parthenotes . Among metabolism genes , P11166 was downregulated , while P15121 , P35354 , O95479 , and P10599 were upregulated in parthenotes , and P22732 did not differ . Among genes involved in compaction/blastulation , P17302 was downregulated in parthenotes , but no differences were detected within P05023 and CDH1 . Within parthenotes , the expression levels of P11166 , TP-1 , and O95479 , and possibly P15121 , resemble patterns described in female embryos . The pro-apoptotic profile is more pronounced in parthenotes than in embryos , which may differ in their way to channel apoptotic stimuli , through p66(shc) and p53 respectively , and in their mechanisms to control pluripotency and de novo methylation . DB05578 ( IMC-1121B ) : Monoclonal antibody inhibition of vascular endothelial growth factor receptor-2 . Angiogenesis , a well-recognized characteristic of malignancy , has been exploited more than any other pathway targeted by biologic anti-neoplastic therapies . Vascular endothelial growth factor receptor-2 ( P35968 ) is the critical receptor involved in malignant angiogenesis with its activation inducing a number of other cellular modifications resulting in tumor growth and metastases . DB05578 ( IMC-1121B ; ImClone Systems Corporation , Branchburg , NJ ) is a fully human monoclonal antibody developed to specifically inhibit P35968 . DB05578 is currently being investigated in multiple clinical trials across a variety of tumor types . Herein , angiogenesis inhibition in cancer is reviewed and up-to-date information on the clinical development of ramucirumab is presented . Design and synthesis of conformationally constrained cyclophilin inhibitors showing a cyclosporin-A phenotype in C. elegans . P62937 ( CypA ) is a member of the immunophilin family of proteins and receptor for the immunosuppressant drug cyclosporin A ( DB00091 ) . Here we describe the design and synthesis of a new class of small-molecule inhibitors for CypA that are based upon a dimedone template . Electrospray mass spectrometry is utilised as an initial screen to quantify the protein affinity of the ligands . Active inhibitors and fluorescently labelled derivatives are then used as chemical probes for investigating the biological role of cyclophilins in the nematode Caenorhabditis elegans . Inhibition of DNA methyltransferase activity prevents tumorigenesis in a mouse model of prostate cancer . Transcriptional silencing of tumor suppressor genes by DNA methylation plays an important role in tumorigenesis . These aberrant epigenetic modifications may be mediated in part by elevated DNA methyltransferase levels . DNA methyltransferase 1 ( P26358 ) , in particular , is overexpressed in many tumor types . Recently , we showed that P26358 is transcriptionally regulated by E2F transcription factors and that retinoblastoma protein ( P06400 ) inactivation induces P26358 . Based on these observations , we investigated regulation of P26358 by polyomavirus oncogenes , which potently inhibit the P06400 pocket protein family . Infection of primary human prostate epithelial cells with BK polyomavirus dramatically induced P26358 transcription following large T antigen ( TAg ) translation and E2F activation . For in vivo study of P26358 regulation , we used the transgenic adenocarcinoma of the mouse prostate ( TRAMP ) model , which expresses the SV40 polyomavirus early region , including TAg , under control of a prostate-specific promoter . Analysis of TRAMP prostate lesions revealed greatly elevated P26358 mRNA and protein levels beginning in prostatic intraepithelial neoplasia and continuing through advanced prostate cancer and metastasis . Interestingly , when TRAMP mice were treated in a chemopreventive manner with the DNA methyltransferase inhibitor DB01262 ( 5-aza ) , 0 of 14 mice developed prostate cancer at 24 weeks of age , whereas 7 of 13 ( 54 % ) control-treated mice developed poorly differentiated prostate cancer . Treatment with 5-aza also prevented the development of lymph node metastases and dramatically extended survival compared with control-treated mice . Taken together , these data suggest that P26358 is rapidly activated by P06400 pathway inactivation , and that DNA methyltransferase activity is required for malignant transformation and tumorigenesis . 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 . 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 . Inhibition of P01375 -induced cyclooxygenase-2 expression by Mycobacterium bovis BCG in human alveolar epithelial A549 cells . P35354 ( P35354 ) is implicated in pathophysiological processes associated with the initiation or maintenance of host inflammatory responses to infection . Our results demonstrates that Mycobacterium bovis BCG ( M. bovis BCG ) downregulates tumour necrosis factor-alpha ( P01375 ) -induced P35354 gene expression in alveolar epithelial cells by inhibiting the phosphorylations of P04049 and p38 kinases . Further , M. bovis BCG-mediated inhibition of P35354 or p38 mitogen-activated protein kinase could be reversed by DB02860 , a selective inhibitor of DB00133 / DB00156 phosphatases . Moreover , M. bovis BCG inhibited the P01375 -triggered NF-kappaB activation following IkappaB degradation . Taken together , these results suggest that the attenuation of P35354 expression by vaccine strain , M. bovis BCG , represents a novel strategy to maintain robust host proinflammatory responses to subsequent challenges with virulent tuberculosis bacilli . Heterogeneous expression of cyclooxygenase-2 and inducible nitric oxide synthase within colorectal tumors : correlation with tumor angiogenesis . BACKGROUND : Recent studies have shown that the cyclooxygenase ( P36551 ) and the inducible nitric oxide synthase ( P35228 ) pathways are involved in the development of tumor angiogenesis in human cancers . AIMS : To investigate whether a different pattern of P35354 and P35228 expression/activity exists within different areas of colorectal tumors and to analyze the relationship between these two enzymes and tumor angiogenesis . METHODS : Microvessel density ( P53602 ) and P35354 , P35228 , vascular endothelial growth factor ( P15692 ) and P15692 receptor-2 ( P35968 ) protein expression were evaluated at both the invasive front ( IF ) and the tumor center ( TC ) in 46 human colorectal cancer specimens . We also investigated the concentration of DB00917 and NO at the same sites . RESULTS : P35354 and P35228 protein expression and activity were significantly higher within the IF than the TC of the tumor specimens . Similarly , P53602 and P15692 / P35968 expression significantly increased from the TC to the IF . Only P35354 expression was significantly correlated with P53602 and P15692 / P35968 expression at both the TC and the IF . CONCLUSION : Our study shows a heterogeneous expression of P35354 and P35228 in colorectal cancer . The up-regulation of P35354 at the IF parallels an increase in vessel density and P15692 / P35968 expression , thus supporting the hypothesis that the tumor periphery is the most aggressive portion of a colorectal tumor . DB08901 -- a step forward in overcoming resistance in chronic myeloid leukemia . With the current therapy , the improvement in survival of patient with early chronic phase chronic myelogenous leukemia ( CML ) is unrivaled by that of any other leukemia . In fact , extrapolation of the survival curves may suggest that life expectancy of patients who achieve and maintain predetermined milestones may not differ from that of the age-matched healthy adults . The main reasons for such success are the presence of a well-defined molecular target , the P11274 - P00519 oncogene , necessary and sufficient for the initiation and propagation of CML , and the powerful and selective agents that inhibit it . Five U.S . Food and Drug Administration ( FDA ) -approved tyrosine kinase inhibitors ( TKI ) , each with unique activities and toxicity profiles , allow for individualized patient care . Despite the remarkable responses of most patients , a small but significant fraction of patients develops clinical resistance to the TKIs , some of which is attributed to the P11274 - P00519 kinase domain mutations affecting TKI binding and activity . The recently approved third-generation TKI ponatinib showed remarkable activity in the patients with multi-TKI-resistant disease . Particularly impressive was its efficacy in patients with T315I mutation that is resistant to all other TKIs . In lieu of the current emphasis on achieving earlier and more profound responses and excellent activity of ponatinib in the refractory setting , its optimal position among the available armamentarium of agents is being established . 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 . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Anti-inflammatory effect of transduced PEP-1-cyclophilin A in Raw264.7 cells and 12-O-tetradecanoylphorbol-13-acetate-induced mice . AIMS : P62937 ( CypA ) is an immunophilin that acts as a receptor for the immunosuppressant drug cyclosporine A ( DB00091 ) . CypA has emerged as a potential drug target for several inflammatory diseases , although the details of its mechanism are unclear . We examined the protective effects of CypA on inflammation in Raw 264.7 cells and animal models . MAIN METHODS : A human CypA gene was fused with a protein transduction domain , PEP-1 peptide , to construct a cell permeable PEP-1-CypA protein . The protein expression level of cyclooxygenase-2 ( P35354 ) and cytokines was detected by Western blot , ELISA and mRNA level of P35354 and cytokines were measured by RT-PCR . The nuclear factor-kappa B ( NF-kB ) and mitogen-activated protein kinase ( MAPK ) activation were analyzed by Western blot and electrophoretic mobility shift assay . Skin inflammation was detected with immunohistochemistry . KEY FINDINGS : Transduced PEP-1-CypA protein markedly inhibited lipopolysaccharide- and 12-O-tetradecanoyl phorbol-13-acetate-induced expression levels of P35354 as well as pro-inflammatory cytokine levels in vitro and in vivo . Furthermore , transduced PEP-1-CypA protein resulted in a significant reduction in the activation of NF-kB and MAPK . SIGNIFICANCE : The results indicate that PEP-1-CypA inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK activation upon stimulation of inflammation in vitro and in vivo . PEP-1-CypA protein may potentially be used as a therapeutic agent against skin diseases-related inflammation . Sublethal concentrations of diverse gold compounds inhibit mammalian cytosolic thioredoxin reductase ( TrxR1 ) . P30044 ( TrxR ) reduces thioredoxin ( P10599 ) , thereby contributing to cellular redox balance , facilitating the synthesis of deoxy-ribose sugars for DNA synthesis , and regulating redox-sensitive gene expression . DB00995 is a gold compound that potently inhibits TrxR . This inhibition is one suspected mechanism of auranofin 's therapeutic benefit in the treatment of rheumatoid arthritis . The use of other gold compounds to treat cancer or inflammatory disease may rely on their ability to inhibit TrxR . In the current study , we tested the hypothesis that a variety of gold compounds may inhibit TrxR . METHODS : We exposed rat-TrxR1 to auranofin , gold sodium thiomalate , sodium aurothiosulfate , triphenyl phosphine gold chloride , or gold acetate , and measured TrxR activity ex vivo . We then compared TrxR1 inhibitory levels of gold compounds to those that inhibited mitochondrial activity of THP1 monocytes and OSC2 epithelial cells , estimated by succinate dehydrogenase activity . RESULTS : All gold compounds inhibited TrxR1 at concentrations ranging from 5 to 4000 nM ( 50 % inhibitory concentration ) . The oxidation state of gold did not correlate with inhibitory potency , but ligand configuration was important . Au(I)-phosphine compounds ( triphenyl phosphine gold chloride and auranofin ) were the most potent inhibitors of TrxR . All TrxR1 inhibitory concentrations were sublethal to mitochondrial activity in both THP1 and OSC2 cells . CONCLUSIONS : Diverse types of gold compounds may be effective inhibitors of TrxR1 at concentrations that do not suppress cellular mitochondrial function . Inhibition may be optimized to some degree by altering the ligand configuration of the compounds . These results support future study of a variety of Au compounds for therapeutic development as inhibitors of TrxR1 . APRIL and Q9Y275 promote increased viability of replicating human B2 cells via mechanism involving cyclooxygenase 2 . Of relevance to both protective and pathogenic responses to Ag is the recent finding that soluble molecules of the innate immune system , i.e. , P05112 , B cell-activation factor of the P01375 family ( Q9Y275 ) , and P01024 , exhibit significant synergy in promoting the clonal expansion of human B2 cells following low-level P11274 ligation . Although P05112 , Q9Y275 , and C3dg each contribute to early cell cycle entry and progression to S phase , only Q9Y275 promotes later sustained viability of progeny needed for continued cycling . The present study sought to further clarify the mechanisms for Q9Y275 's multiple functions . By comparing Q9Y275 and a proliferation-inducing ligand ( APRIL ) efficacy at different stages in the response ( only Q9Y275 binds Q96RJ3 ; both bind transmembrane activator and calcium modulator and cyclophilin ligand interactor ( O14836 ) and B cell maturation Ag , the early role was attributed to Q96RJ3 , while the later role was attributed to O14836 /B cell maturation Ag . Importantly , Q9Y275 - and APRIL-promoted viability of cycling lymphoblasts was associated with sustained expression of cyclooxygenase 2 ( P35354 ) , the rate-limiting enzyme for DB00917 synthesis , within replicating cells . Supernatants of cultures with Q9Y275 and APRIL contained elevated DB00917 . Although P35354 inhibitors diminished daughter cell viability , exogenous DB00917 ( 1-1000 nM ) increased the viability and recovery of lymphoblasts . Increased yield of viable progeny was associated with elevated Mcl-1 , suggesting that a Q9Y275 /APRIL --> O14836 --> P35354 --> DB00917 --> Mcl-1 pathway reduces activation-related , mitochondrial apoptosis in replicating human B2 cell clones .

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interacts_with DB00391?

multiple_choice

Identification and quantification of dopamine receptor 2 in human eutopic and ectopic endometrium : a novel molecular target for endometriosis therapy . Previous studies in an experimental mouse model of endometriosis have shown that the dopamine agonist ( DA ) cabergoline ( Cb2 ) reduces angiogenesis and endometriotic lesions , hypothetically binding to the dopamine receptor type-2 ( P14416 ) . To date , this has not been described in human endometrium and/or endometriotic lesions . Thus , we aimed to investigate the presence of P14416 in said tissues . Endometrium fragments were implanted in nude mice treated with different doses of Cb2 . Polymerase chain reaction assays and immunohistochemistry were performed to analyze the gene and protein expressions ( respectively ) of P14416 , P15692 , and P15692 receptor-2 ( P35968 ) . In addition , lesions and endometrium from women with mild and severe endometriosis and endometrium from healthy women were collected to analyze their gene expression profile . In experimental endometriosis , P14416 was expressed at gene and protein levels in all three groups . P15692 gene and protein expressions were significantly lower in lesions treated with Cb2 than in controls . P35968 protein expression was significantly lower in experimental lesions treated with Cb2 than in controls . In eutopic endometria , there was a significant decrease in P14416 expression and an increase in P15692 in women with mild and severe endometriosis with respect to healthy patients . In endometriosis , P35968 expression was significantly higher in red than in white and black lesions . P15692 expression was significantly lower in black than in red lesions . P14416 is present in the human eutopic and ectopic endometrium and is regulated by DA , which provides the rationale for pilot studies to explore its use in the treatment of endometriosis . Analyses of cross species polymerase chain reaction products to infer the ancestral state of human polymorphisms . In numerous population genetic and disease association studies decisions about the ancestry of polymorphic alleles are often made based on the relative frequency of the alleles in the extant populations with the most frequent allele being deemed as ancestral . However , the frequency of an allele in a population is generally not a perfect indicator of its ancestral status . A more accurate method to assess ancestral/derived status of polymorphic alleles involves identification of shared alleles between species . We used this strategy to examine genomic regions homologous to several human polymorphisms in four species of non-human primates . Cross species polymerase chain reaction ( CS-PCR ) , with primers designed from human sequence , was used to investigate regions of interest . Nineteen polymorphisms at six loci ( P14416 , HOXB@ , PAH , D4S10 , P10745 , and P07949 ) were examined either by restriction fragment length analysis of PCR products ( PCR-RFLP ) or by direct sequencing . At seventeen of the eighteen PCR-RFLPs , non-human primates were monomorphic and identical to each other for either lack of restriction enzyme site or presence of the site . Thus , at these seventeen polymorphic sites the shared alleles are most likely to be the ancestral ones in humans . In several cases we have used sequence data to further demonstrate that the nucleotide at the site of the polymorphism is conserved between species confirming the hypothesis of a single ancestral allele . However , not all human alleles can be simply resolved into ancestral and derived ; sequence data from one PCR-RFLP ( in an intron of the PAH locus ) and a single strand conformational polymorphism ( SSCP ) in the 3' untranslated region ( UTR ) of the P14416 gene illustrate this point . 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 . Neurohypophysial hormone secretion in hyperprolactinaemic women . P01236 ( PRL ) has been reported to promote antidiuresis and increase intestinal water-electrolyte absorption , whereas osmolar changes have been shown to influence PRL secretion . However , the mechanisms of action of PRL on the salt-water balance remain unclarified . The present clinical study targeted the effects of hyperprolactinaemia on the secretion of arginine-8-vasopressin ( AVP ) , oxytocin ( P01178 ) and cortisol . Plasma AVP and P01178 were measured by radioimmunoassay , and cortisol by fluorimetry . In healthy women ( 21-39 y , n=6 ) , an oral water load ( OWL , 20 ml/bw ) significantly suppressed the plasma levels of AVP , P01178 and cortisol , and the PRL level too tended to decrease . In hyperprolactinaemic females ( 22-41 y , n=6 , three with pituitary adenomas ) , water retention was registered following an OWL , together with paradoxical AVP and P01178 level increases , whereas the cortisol response remained normal , and the PRL level did not change at all . DB11320 ( 0.5 mg sc ) stimulated the release of AVP , P01178 and cortisol in the control and hyperprolactinaemic groups alike . These data suggest that alterations in AVP and P01178 hypersecretion may contribute to the water retention in hyperprolactinaemia . 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 . 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 . Dopamine-related genes and their relationships to monoamine metabolites in P04141 . Monoamine metabolite ( MM ) levels in lumbar cerebrospinal fluid ( P04141 ) are extensively used as indirect estimates of monoamine turnover in the brain . In this study we investigated genotypes for DNA polymorphisms in the D2 ( P14416 ) , D3 ( P35462 ) , and D4 ( P21917 ) dopamine receptor and tyrosine hydroxylase ( TH ) genes and their relationships to P04141 MM in healthy volunteers ( n = 66 ) . Concentrations of homovanillic acid ( HVA ) , 3-methoxy-4-hydroxyphenylglycol ( MHPG ) , and 5-hydroxyindoleacetic acid ( 5-HIAA ) were corrected for back length , a confounding variable . Corrected MM levels were not related to age , gender , height , weight heredity , season or atmospheric pressure at sampling . Individuals with specific P14416 and TH allele and genotype configurations significantly differed in HVA and MHPG concentrations . P35462 homo- and heterozygotic genotypes had significantly different P04141 5-HIAA levels . P21917 genotypes were not related to MM concentrations . The results suggest that specific P14416 , P35462 , and TH genotypes participate in the regulation of monoamine turnover in the central nervous system . Accordingly monoamine receptors and synthesizing enzyme genotypes appear to be variance factors influencing MM concentrations in P04141 . The relationships found in this study support MM concentrations as markers for monoamine transmission in the human brain . Paragangliomas/Pheochromocytomas : clinically oriented genetic testing . Paragangliomas are rare neuroendocrine tumors that arise in the sympathetic or parasympathetic nervous system . Sympathetic paragangliomas are mainly found in the adrenal medulla ( designated pheochromocytomas ) but may also have a thoracic , abdominal , or pelvic localization . Parasympathetic paragangliomas are generally located at the head or neck . Knowledge concerning the familial forms of paragangliomas has greatly improved in recent years . Additionally to the genes involved in the classical syndromic forms : P40337 gene ( von Hippel-Lindau ) , P07949 gene ( Multiple Endocrine Neoplasia type 2 ) , and P21359 gene ( Neurofibromatosis type 1 ) , 10 novel genes have so far been implicated in the occurrence of paragangliomas/pheochromocytomas : P31040 , P21912 , Q99643 , O14521 , Q9NX18 , O75204 , P61244 , Q9GZT9 , Q99814 , and O60333 . It is currently accepted that about 35 % of the paragangliomas cases are due to germline mutations in one of these genes . Furthermore , somatic mutations of P07949 , P40337 , P21359 , P61244 , Q99814 , and H- DB01367 can also be detected . The identification of the mutation responsible for the paraganglioma/pheochromocytoma phenotype in a patient may be crucial in determining the treatment and allowing specific follow-up guidelines , ultimately leading to a better prognosis . Herein , we summarize the most relevant aspects regarding the genetics and clinical aspects of the syndromic and nonsyndromic forms of pheochromocytoma/paraganglioma aiming to provide an algorithm for genetic testing . 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 . 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 . 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 . Direct anti-inflammatory mechanisms contribute to attenuation of experimental allograft arteriosclerosis by statins . BACKGROUND : Despite the development of effective immunosuppressive therapy , transplant graft arterial disease ( Q99259 ) remains the major limitation to long-term graft survival . The interplay between host inflammatory cells and donor vascular wall cells results in an intimal hyperplastic lesion , which leads to ischemia and graft failure . P04035 inhibitors ( statins ) reduce Q99259 in human cardiac allografts , although it is unclear whether this is secondary to cholesterol lowering or other mechanisms . This study tested the hypothesis that statins can suppress Q99259 by cholesterol-independent pathways . METHODS AND RESULTS : We performed heterotopic murine cardiac transplants in total allogeneic or major histocompatibility complex class II-mismatched combinations . Transplanted animals received either control chow , chow containing 25 ppm cerivastatin ( low dose ) , or chow containing 125 ppm cerivastatin ( high dose ) . Mean plasma cerivastatin concentrations were 0.0 ( control ) , 10.1 ( low dose ) , and 21.9 ( high dose ) nmol/L , respectively . Plasma cholesterol levels were the same in all groups . Q99259 scores decreased in low-dose ( P < 0.05 ) and high-dose ( P < 0.0001 ) cerivastatin groups compared with controls , with concomitant reduction in graft-infiltrating cells and significantly decreased intragraft RANTES and monocyte chemotactic protein-1 mRNA expression . DB00439 , as well as other statins , also reduced RANTES and monocyte chemotactic protein-1 production in mouse endothelial cells stimulated with interferon-gamma and tumor necrosis factor-alpha in vitro . CONCLUSIONS : Clinically achievable levels of an P04035 inhibitor attenuate Q99259 in murine heart transplants , diminish host inflammatory cell recruitment , and do not alter cholesterol levels . These results indicate that statins can affect arterial biology and inflammation independently of their effects on cholesterol metabolism . Role of estrogen receptors in menstrual cycle-related neoangiogenesis and their influence on endothelial progenitor cell physiology . OBJECTIVE : To study whether estrogen receptors ( ERs ) are expressed in vitro and in vivo by female circulating endothelial progenitor cells ( EPCs ) ; and the role of ERs in the periodic vascular damage and repair that occurs during the menstrual cycle . DESIGN : Quantification of circulating progenitor cells , EPCs , and relative P61073 + fraction by flow cytometry . Quantification of plasma 17beta-E(2) by electrochemiluminescent immunoassay . Expression of ERs by immunofluorescence and immunohistochemistry . P03372 , P61073 , and matrix metalloproteinase 9 gene expression by reverse transcriptase-polymerase chain reaction and real-time polymerase chain reaction . SETTING : University clinic and academic research laboratory . PATIENT(S) : Twelve young fertile women ( aged 22-27 years ) observed for 6 months , 10 postmenopausal women ( aged 52-63 years ) , and 50 male control subjects ( aged 24-61 years ) . INTERVENTION(S) : Blood ( 35 mL ) was collected at each observation point . MAIN OUTCOME MEASURE(S) : Correlation between 17beta-E(2) exposure and neoangiogenesis markers . RESULT(S) : Estrogen receptors are expressed both in cultured EPCs after prolonged estrogen stimulation and in circulating EPCs , such as in P28906 + cells in bone marrow . The number of Q92731 + and P61073 + EPCs increased during the ovulatory phase , and this increase is probably mediated by Q92731 and matrix metalloproteinase 9 . CONCLUSION(S) : DB00286 play a key role in neoangiogenesis processes , such as endometrium recovery , and this mechanism involves both a central action ( on bone marrow ) and a cytokine-mediated peripheral one ( on endothelium ) . Differential expression of the alternatively spliced P35372 isoform μ-opioid receptor-1K in HIV-infected individuals . OBJECTIVE : We previously examined the expression of specific C-terminal μ-opioid receptor ( MOR ) splice variants in human central nervous system cell types and HIV-infected brain tissue from individuals with neurocognitive impairment ± HIV encephalitis ( HIVE ) . In the present study , we examined the N-terminal splice variant MOR-1K , which mediates excitatory cellular signaling . METHODS AND RESULTS : We found segregation of expression ranging from undetectable to seemingly exclusive across nervous system cell types compared to the pool of C-terminal MOR splice variants using the real-time polymerase chain reaction ( RT-PCR ) . Expression of MOR-1K mRNA was also increased in HIV-infected individuals with combined neurocognitive impairment and HIVE compared with the other groups . MOR-1K expression correlated with the level of patient neurocognitive impairment , whereas the pool of C-terminal MOR splice variants did not . HIVE was also associated with increased expression of the inflammatory mediators P13500 , P80075 , and RANTES , but not the host HIV coreceptors P61073 and P51681 or the P01730 receptor using qRT-PCR . Network analysis of microarray data from these same patients revealed filamin A ( P21333 ) as a possible interaction partner with MOR-1K , and P21333 gene expression was also found to be upregulated in HIVE using qRT-PCR . Overexpression of P21333 in HEK293 cells redistributed MOR-1K from intracellular compartments to the cell surface . CONCLUSION : These results suggest that HIVE , and neurocognitive impairment depending on its severity , are associated with enhanced MOR-1K signaling through both increased expression and trafficking to the cell surface , which may alter the contribution of MOR receptor isoforms and exacerbate the effects of MOR activation in neuroAIDS . Dopamine receptors and psychiatric drug treatment . The established antipsychotic drugs act mainly by antagonizing dopamine mediated synaptic transmission in the brain . Increase in the rate of production of dopamine metabolites as well as the firing rate of dopamine-containing neurons can be interpreted as compensatory responses to an interruption of synaptic transmission at dopamine nerve terminals . The demonstration of involvement of limbic and cortical mechanisms in the antipsychotic activity of neuroleptic drugs is far more difficult than the involvement of nigro-striatal and tubero-infundibular mechanisms in the neurological and neuroendocrine effects of these drugs . Application of radioreceptor techniques to dopamine research has supported the findings obtained by other neuropsychopharmacological research techniques , providing more direct evidence of dopamine receptor blockade by neuroleptic drugs . Further research is needed especially in studying the nature of the time-dependent adaptive changes at the receptor sites as well as the differences between the different dopamine projections and neural systems in the brain . The different subtypes of dopamine receptors in the brain , currently called D1 and D2 dopamine receptors , seem to be parallel , although in many respects independently-acting regulatory systems . P14416 -selective antagonists such as sulpiride seem to cause selective D2 receptor up-regulation . P01236 secretion seems to be regulated by D2 dopamine receptors . The exact physiological role of D1 dopamine receptors as well as the clinical consequences of selective D1 antagonism is not known . DB00391 and clozapine are examples of atypical neuroleptic compounds that have quite different profile of action , the former having strong and selective antidopaminergic action , the latter combining a number of non- dopaminergic mechanisms with rather slight effects on dopamine receptors. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB08626 -induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer 's disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 and cortical Abeta40 that were 147 and 142 % of the control group , respectively . Results for Abeta42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD . P01236 influence on cytosol and nuclear androgen receptors in the ventral , dorsal , and lateral lobes of the rat prostate . PRL augments testosterone-mediated growth of the prostate in a permissive manner . To elucidate the mechanism of this hormonal interaction , the present study examined the effect of PRL on cytosol and nuclear androgen receptors in the three prostate lobes . Adult male Sprague-Dawley rats were castrated , given 5-mm Silastic implants of testosterone , and either grafted with two anterior pituitary glands under the kidney capsule or sham operated . Three weeks later , animals were killed , serum was collected for PRL and testosterone RIA , and the ventral , dorsal , and lateral prostate lobes were processed for either cytosol or nuclear androgen receptor quantitation . Pituitary grafts significantly elevated the serum PRL concentration and increased the weight and the content of protein and DNA of the lateral prostate lobe compared to control values . There was no effect on these parameters in the ventral or dorsal lobes . P10275 levels and apparent distributions were different in the ventral , dorsal , and lateral lobes of control animals . Unoccupied and total cytosolic androgen receptors in the three separate prostate lobes were not significantly affected by the presence of the grafts . However , an elevated PRL concentration was associated with an increase ( P less than 0.005 ) in nuclear androgen receptor content in the lateral lobe exclusively . The binding affinity was not altered by pituitary grafts in any of the lobes . These findings suggest that PRL promotes lateral prostatic growth by increasing nuclear androgen receptor levels in that tissue and , thus , optimizes its response to circulating testosterone . Polymorphisms of dopamine receptor/transporter genes and risk of non-small cell lung cancer . BACKGROUND : The dopaminergic pathway may be of interest in assessing risk of non-small cell lung cancer ( NSCLC ) . Dopamine receptors are expressed in alveolar epithelial cells and human lung tumours , and dopamine inhibits both cell proliferation in vitro and growth of lung tumour xenografts in nude mice . Moreover , dopamine selectively inhibits the vascular permeability and angiogenic activity of vascular endothelial growth factor ( P15692 / P15692 ) . The bioavailability of dopamine is regulated by dopamine receptors D2 ( P14416 ) , D4 ( P21917 ) and dopamine transporter 1 ( Q01959 / Q01959 ) genes . METHODS : We have analysed 10 single nucleotide polymorphisms in P14416 , P21917 and Q01959 / Q01959 genes in relation to lung cancer risk in a case-control study of smoking subjects . The study subjects were 413 healthy individuals from general population and 335 NSCLC cases . Both cases and controls were Caucasians of Norwegian origin . RESULTS : We demonstrate that P14416 polymorphisms -141Cdel , 3208G > T , TaqIB ; P21917 -521C > T and Q01959 / Q01959 -1476T > G are associated with a two- to five-fold increased NSCLC risk . The variant alleles of P14416 1412A > G and 960C > G had protective effects . CONCLUSION : The dopamine receptor/transport gene polymorphisms are associated with the risk of NSCLC among smokers . The data show that the polymorphisms resulting in lower dopamine bioavailability were associated with increased risk of NSCLC . Comorbid migraine with aura , anxiety , and depression is associated with dopamine D2 receptor ( P14416 ) NcoI alleles . BACKGROUND : Unrelated individuals ( n = 242 ) were interviewed directly for the presence of migraine , anxiety disorders , and major depression . MATERIALS AND METHODS : The data described in this study are derived from a clinical genetic relational database that was developed initially for the genetic analysis of migraine . Genotyping of the P14416 NcoI C to T polymorphism located in exon 6 ( His313His ) was performed using previously described primers . RESULTS : A significantly increased incidence of migraine with aura ( MWA ) , major depression , generalized anxiety disorder ( Q99259 ) , panic attacks , and phobia was observed in individuals with the P14416 NcoI C/C genotype compared with individuals with an P14416 NcoI T allele . Specifically , 69 % ( 91/131 ) of P14416 NcoI C/C individuals in the present study met criteria for at least one of these neuropsychiatric disorders versus only 22 % ( 4/18 ) of the P14416 NcoI T/T individuals ( Chi-square = 15.29 ; p < 0.00005 ) . The P14416 NcoI C allele frequency is significantly higher ( Chi-square = 17.13 ; p < 0.00002 ) in individuals with MWA , anxiety disorders , and/or major depression ( C allele frequency = 0.80 ) than in individuals who have none of these disorders ( C allele frequency = 0.67 ) . CONCLUSIONS : These data indicate that MWA , anxiety disorders , and major depression can be components of a distinct clinical syndrome associated with allelic variations within the P14416 gene . Clinical recognition of this genetically based syndrome has significant diagnostic and therapeutic implications . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Systemic administration of oxytocin reduces basal and lipopolysaccharide-induced ghrelin levels in healthy men . DB00107 ( P01178 ) and ghrelin have several common properties such as the involvement in the first phase response to stressors , in appetite regulation , and in the modulation of neural functions . Despite a recent study showing that intraventricular administration of ghrelin activates P01178 neurons , little is known on the cross-talk between these two peptides . Here , we investigated the role of the i.v. administration of P01178 on circulating ghrelin concentrations under fasting conditions and during the lipopolysaccharide ( LPS ) -induced endotoxemia . A randomized placebo-controlled cross-over study was performed in ten healthy men . In four study sessions , the participants received once placebo , once P01178 ( 1 pmol/kg per min over 90 min ) , once LPS ( 2 ng/kg ) , and once both P01178 and LPS . Plasma ghrelin , glucose , and free fatty acid ( FFA ) levels were measured at regular intervals during the first 6 h following the LPS bolus . Systemic administration of P01178 decreased within 1 h plasma ghrelin levels ( 611+/-54 vs 697+/-52 pg/ml in placebo days , P=0.013 ) and increased plasma glucose and FFA concentrations ( P=0.002 and P=0.005 respectively ) . P01178 also reduced the LPS-induced surge in ghrelin at time point 2 h ( P=0.021 ) . In summary , i.v. administration of P01178 decreases circulating levels of ghrelin during fasting , as well as following LPS-induced endotoxemia in healthy men . The cross-talk between P01178 and ghrelin might be important in the regulation of energy homeostasis and stress responses .

MH_train_1412

MH_train_1412

MH_train_1412

interacts_with DB01151?

multiple_choice

Regulation of Q68DA7 subdomain interactions and function in neuronal nitric oxide synthase . DB00435 synthases ( NOS ) are modular , calmodulin- ( P62158 - ) dependent , flavoheme enzymes that catalyze oxidation of l-arginine to generate nitric oxide ( NO ) and citrulline . During catalysis , the Q68DA7 subdomain cycles between interaction with an NADPH- DB03147 subdomain to receive electrons and interaction with an oxygenase domain to deliver electrons to the NOS heme . This process can be described by a three-state , two-equilibrium model for the conformation of the Q68DA7 subdomain , in which it exists in two distinct bound states ( Q68DA7 -shielded ) and one common unbound state ( Q68DA7 -deshielded ) . We studied how each partner subdomain , the Q68DA7 redox state , and P62158 binding may regulate the conformational equilibria of the Q68DA7 module in rat neuronal NOS ( P29475 ) . We utilized four P29475 protein constructs of different subdomain composition , including the isolated Q68DA7 subdomain , and determined changes in the conformational state by measuring the degree of Q68DA7 shielding by fluorescence , electron paramagnetic resonance , or stopped-flow spectroscopic techniques . Our results suggest the following : ( i ) The NADPH- DB03147 subdomain has a far greater capacity to interact with the Q68DA7 subdomain than does the oxygenase domain . ( ii ) P62158 binding has no direct effects on the Q68DA7 subdomain . ( iii ) P62158 destabilizes interaction of the Q68DA7 subdomain with the NADPH- DB03147 subdomain but does not measurably increase its interaction with the oxygenase domain . Our results imply that a different set point and P62158 regulation exists for either conformational equilibrium of the Q68DA7 subdomain . This helps to explain the unique electron transfer and catalytic behaviors of P29475 , relative to other dual-flavin enzymes . The aptamer DB05202 is a potent and specific inhibitor of von Willebrand Factor mediated ex vivo platelet function in acute myocardial infarction . DB05202 is an aptamer , which blocks binding of the von Willebrand Factor ( P04275 ) A1 domain to platelet GPIb receptors . P04275 is increased in the elderly an in the setting of acute myocardial infarction ( AMI ) , as reflected by increased shear-dependent platelet function . We hypothesized that DB05202 concentration-dependently inhibits ex vivo platelet function , and that this concentration effect relationship may be shifted in patients with AMI . We studied ex vivo dose response curves for DB05202 on P04275 activity , shear-dependent platelet function , and agonist-induced platelet aggregation . We included patients with AMI on standard treatment ( n = 40 ) , young ( n = 20 ) and elderly controls ( n = 20 ) in this ex vivo dosing study . AMI patients displayed approximately 2-fold increased plasma levels of P04275 activity as compared to controls . DB05202 inhibited P04275 activity ( IC90 : approximately 3-4 microg/mL ) and shear dependent platelet function ( Platelet Function Analyzer ( PFA-100 ) , IC50 : approximately 0.5-0.9 microg/mL and Cone and Plate Analyzer ( P15085 ) , IC50 : approximately 0.1-0.4 microg/mL in citrated blood ) at comparable concentrations in all groups . In contrast to P08514 /IIIa antagonists , DB05202 did not inhibit platelet aggregation induced by ADP , collagen or arachidonic acid at concentrations ( 10 microg/mL ) that fully inhibited P04275 dependent platelet function . DB05202 potently and specifically inhibits P04275 activity and P04275 dependent platelet function , even in the setting of AMI where P04275 activity is increased . 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 . DB06268 ( Q9Y6W8 -Texas Biotechnology ) . Q9Y6W8 -Texas Biotechnology is developing the endothelin A ( P25101 ) receptor antagonist , sitaxsentan , for the potential treatment of pulmonary hypertension , congestive heart failure ( CHF ) , chronic obstructive pulmonary disease and subarachnoid hemorrhage [ 205713 ] , [ 302200 ] . The compound is in phase IIa trials as an iv formulation for CHF and has completed phase I safety trials as an oral formulation [ 272071 ] . Phase II/III trials for pulmonary hypertension are planned for the first quarter of 2001 [ 3945711 ] . In June 2000 , Q9Y6W8 and Texas Biotechnology established a joint venture to develop and commercialize endothelin antagonists [ 370007 ] . US-05591761 was issued to Texas in January 1997 , covering TBC-11251 and several related isomers [ 2309301 . Treatment with DB06603 induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells . Increased levels of misfolded polypeptides in the endoplasmic reticulum ( ER ) triggers the dissociation of glucose-regulated protein 78 ( P11021 ) from the three transmembrane ER-stress mediators , i.e. , protein kinase RNA-like ER kinase ( Q9NZJ5 ) , activating transcription factor-6 ( P18850 ) , and inositol-requiring enzyme 1alpha , which results in the adaptive unfolded protein response ( UPR ) . In the present studies , we determined that histone deacetylase-6 ( Q9UBN7 ) binds and deacetylates P11021 . Following treatment with the pan-histone deacetylase inhibitor DB06603 ( Novartis Pharmaceuticals ) , or knockdown of Q9UBN7 by short hairpin RNA , P11021 is acetylated in 11 lysine residues , which dissociates P11021 from Q9NZJ5 . This is associated with the activation of a lethal UPR in human breast cancer cells . Coimmunoprecipitation studies showed that binding of Q9UBN7 to P11021 requires the second catalytic and COOH-terminal BUZ domains of Q9UBN7 . Treatment with DB06603 increased the levels of phosphorylated-eukaryotic translation initiation factor ( p-eIF2alpha ) , P18848 , and CAAT/enhancer binding protein homologous protein ( P35638 ) . DB06603 treatment also increased the proapoptotic Q13323 , O43521 , Q07812 , and Q16611 levels , as well as increased the activity of caspase-7 . Knockdown of P11021 sensitized MCF-7 cells to bortezomib and DB06603 -induced UPR and cell death . These findings indicate that enforced acetylation and decreased binding of P11021 to Q9NZJ5 is mechanistically linked to DB06603 -induced UPR and cell death of breast cancer cells . Mol Cancer Ther ; 9(4) ; 942-52 . (c)2010 AACR . Stimulation of cloned human serotonin P28221 beta receptor sites in stably transfected P13671 glial cells promotes cell growth . The involvement of serotonin P28221 beta receptor sites was investigated in the growth of rat P13671 glial cells permanently transfected with a gene encoding a human P28221 beta receptor . The 5-HT receptor identity of control and transfected P13671 glial/ P28221 beta cells was determined by reverse transcription-polymerase chain reaction using primers specific for rat P08908 , rat P28222 , rat P28221 alpha , human P28221 beta , and rat 5- Q13049 receptor genes . Constitutive mRNA for 5- Q13049 receptors was present in control and transfected P13671 glial/ P28221 beta cells , whereas mRNA for P28221 beta receptor sites was only present in the transfected P13671 glial/ P28221 beta cell line . 5-HT inhibited forskolin-stimulated cyclic AMP formation and promoted cell growth , in contrast to the absence of any measurable effect in pcDNA3 plasmid-transfected and nontransfected P13671 glial cells . The 5-HT effects could be mimicked by sumatriptan ( EC50 = 44-76 nM ) and were totally and partially blocked by methiothepin ( IC50 = 9 nM ) and GR 127,935 ( 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol-3-yl)-biphenyl-4-carbox yli c acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide ; IC50 = 97 pM ) , respectively . No effect on cell growth was measured with the 5-HT2 receptor agonist DOI [ 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ; 10 microM ] , suggesting that 5- Q13049 receptors are not involved in the 5-HT-stimulated P13671 glial/ P28221 beta cell growth . Dibutyryl-cyclic AMP ( 0.3 mM ) -treated cultures did not show sumatriptan-promoted cell growth , indicating an inhibitory role for cyclic AMP in the cell growth mediated by P28221 beta receptor sites. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . 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 ) . The squalestatins : novel inhibitors of squalene synthase . Enzyme inhibitory activities and in vivo evaluation of C1-modified analogues . Squalestatin analogues modified in the C1 side chain were prepared and evaluated for their ability to inhibit rat liver microsomal and Candida squalene synthase ( P37268 ) in vitro . While maintaining the 4,6-dimethyloctenoate or 4,6-dimethyloctanoate ester groups at P13671 , a number of modifications to the C1 side chain were well tolerated . However , in the absence of the P13671 ester group , similar modifications to the C1 side chain caused substantial loss of activity . Compounds were also evaluated for their ability to inhibit cholesterol biosynthesis in vivo in rats and to reduce serum cholesterol levels in marmosets . These studies revealed that compounds with similar P37268 inhibitory activities can possess different in vivo durations of action and lipid-lowering abilities . 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 . 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 . 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 . Differential involvement of Galpha12 and Galpha13 in receptor-mediated stress fiber formation . The ubiquitously expressed heterotrimeric guanine nucleotide-binding proteins ( G-proteins ) G12 and Q99941 have been shown to activate the small GTPase Rho . Rho stimulation leads to a rapid remodeling of the actin cytoskeleton and subsequent stress fiber formation . We investigated the involvement of G12 or Q99941 in stress fiber formation induced through a variety of Gq/ P49842 -coupled receptors . Using fibroblast cell lines derived from wild-type and Galphaq/Galpha11-deficient mice , we show that agonist-dependent activation of the endogenous receptors for thrombin or lysophosphatidic acid and of the heterologously expressed bradykinin B2 , vasopressin V1A , endothelin P25101 , and serotonin P28335 receptors induced stress fiber formation in either the presence or absence of Galphaq/Galpha11 . Stress fiber assembly induced through the muscarinic M1 and the metabotropic glutamate subtype 1alpha receptors was dependent on Gq/ P49842 proteins . The activation of the Gq/ P49842 -coupled endothelin ETB and angiotensin AT1A receptors failed to induce stress fiber formation . Lysophosphatidic acid , B2 , and P28335 receptor-mediated stress fiber formation was dependent on Galpha13 and involved epidermal growth factor ( P01133 ) receptors , whereas thrombin , P25101 , and V1A receptors induced stress fiber accumulation via Galpha12 in an P01133 receptor-independent manner . Our data demonstrate that many Gq/ P49842 -coupled receptors induce stress fiber assembly in the absence of Galphaq and Galpha11 and that this involves either a Galpha12 or a Galpha13/ P01133 receptor-mediated pathway . 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 . P37268 inhibition : a novel target for the management of dyslipidemia . A new class of compounds , known as squalene synthase inhibitors , has recently reached phase III clinical trials and may provide another therapeutic option for clinicians to improve risk management of low-density lipoprotein cholesterol ( LDL-C ) . The clinical need for another LDL-C-lowering therapy is evident by the inability to achieve an LDL-C target of less than 70 mg/dL in the majority of very high-risk patients on statin monotherapy . Human clinical trial data with DB05317 , a novel and potent inhibitor of squalene synthase , have not yet been published . Cocaine increases endoplasmic reticulum stress protein expression in striatal neurons . Cocaine administration upregulates the levels of extracellular glutamate and dopamine in the striatum . Activation of the receptors alters calcium homeostasis in striatal neurons leading to the expression of the endoplasmic reticulum ( ER ) stress proteins . It was therefore hypothesized that cocaine upregulates the expression of the ER stress proteins , immunoglobulin heavy chain binding protein ( P11021 ) , Ire1alpha and perk via glutamate and dopamine receptor activation . A novel glutamate microbiosensor and Western immunoblot analyses were mainly performed to test the hypothesis in the rat dorsal striatum . The results showed that i.p. injection of repeated cocaine ( 20 mg/kg ) for nine consecutive days significantly increased extracellular glutamate levels while acute cocaine injection did not . However , the immunoreactivities ( IR ) of the ER stress proteins in the dorsal striatum were significantly increased by either acute or repeated cocaine injections as compared with saline controls . Intrastriatal injection ( i.s. ) of the selective group I metabotropic glutamate receptor ( mGluR ) antagonist N-phenyl-7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxamide ( PHCCC ; 25 nmol ) or the P41594 subtype antagonist 2-methyl-6-(phenylethynyl)pyridine hydrochloride ( MPEP ; 2 and 25 nmol ) significantly decreased repeated cocaine-induced increases in the IR of the ER stress proteins in the injected dorsal striatum . Similarly , the selective D1 antagonist ( R ) -(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride ( SCH23390 ; 0.1 mg/kg , i.p. ) or the N-methyl-d-aspartate antagonist dizocilpine/(5S,10R)-(+)-5-methyl-10,11-dihydro-5H-ibenzo[a,d]cyclohepten-5,10-imine maleate ( MK801 ; 2 nmol , i.s. ) decreased acute or repeated cocaine-induced the IR of the ER stress proteins in the dorsal striatum . These data suggest that cocaine upregulates expression of the ER stress proteins in striatal neurons via a mechanism involving activation of glutamate and dopamine receptors . 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 . 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 . 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 ) Expression of metabotropic glutamate receptors in rat meningeal and brain microvasculature and choroid plexus . This study investigated the distribution of metabotropic glutamate receptors ( mGluRs ) in meningeal and parenchymal microvasculature and in choroid plexus by means of Western blot analysis and immunohistochemistry . Western blot analysis demonstrated mGluR expression in both rat and human leptomeningeal tissues . In the rat , mGluR expression was developmentally regulated , with only Q14416 /3 showing expression at the embryonic day 19 developmental stage . In contrast , Q13255 alpha , Q14416 /3 , mGluR4a , and Q14831 were expressed in leptomeninges from adult rats . Immunohistochemical analyses showed intense Q13255 alpha immunoreactivity in the pia mater and blood vessels in the subarachnoid space and in the arachnoid layer of the meninges . Q14416 /3 , mGluR4a , P41594 , and Q14831 were also expressed in meningeal microvasculature . In addition , the parenchymal microvasculature and choroid plexus were strongly immunoreactive for Q13255 alpha , Q14416 /3 , mGluR4a , P41594 , and Q14831 . We used antibodies specific for phenotypic markers of microvascular and glial cells to characterize the cell type(s) immunopositive for mGluRs . Comparison of staining with anti- P04275 antibody and anti-mGluR antibodies revealed that mGluR immunoreactivity was present in cells that surrounded the luminal surface labeled by the endothelial cell marker . In these cells , smooth muscle actin and mGluR immunoreactivity overlapped , suggesting that , in addition to endothelial cells , pericytes within the microvasculature also express mGluRs . Furthermore , expression of Q13255 alpha was also observed in pure pericyte cultures isolated from bovine retina . These data suggest that glutamate by means of activation of mGluRs may have a broad sphere of physiological influence in the brain which in addition to modulating synaptic transmission may also have a role in determining microvascular function and dysfunction .

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interacts_with DB08907?

multiple_choice

Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Regulation of lipogenesis by cyclin-dependent kinase 8-mediated control of P36956 . Altered lipid metabolism underlies several major human diseases , including obesity and type 2 diabetes . However , lipid metabolism pathophysiology remains poorly understood at the molecular level . P01308 is the primary stimulator of hepatic lipogenesis through activation of the SREBP-1c transcription factor . Here we identified cyclin-dependent kinase 8 ( P49336 ) and its regulatory partner cyclin C ( CycC ) as negative regulators of the lipogenic pathway in Drosophila , mammalian hepatocytes , and mouse liver . The inhibitory effect of P49336 and CycC on de novo lipogenesis was mediated through P49336 phosphorylation of nuclear SREBP-1c at a conserved threonine residue . Phosphorylation by P49336 enhanced SREBP-1c ubiquitination and protein degradation . Importantly , consistent with the physiologic regulation of lipid biosynthesis , P49336 and CycC proteins were rapidly downregulated by feeding and insulin , resulting in decreased SREBP-1c phosphorylation . Moreover , overexpression of CycC efficiently suppressed insulin and feeding-induced lipogenic gene expression . Taken together , these results demonstrate that P49336 and CycC function as evolutionarily conserved components of the insulin signaling pathway in regulating lipid homeostasis . Estradiol increases cell growth in human astrocytoma cell lines through ERα activation and its interaction with Q15788 and Q9Y6Q9 coactivators . Estradiol ( E2 ) regulates several cellular functions through the interaction with estrogen receptor subtypes , ERα and ERβ , which present different functional and regulation properties . ER subtypes have been identified in human astrocytomas , the most common and aggressive primary brain tumors . We studied the role of ER subtypes in cell growth of two human astrocytoma cell lines derived from tumors of different evolution grades : U373 and D54 ( grades III and IV , respectively ) . E2 significantly increased the number of cells in both lines and the co-administration with an ER antagonist ( ICI 182 , 780 ) significantly blocked E2 effects . ERα was the predominant subtype in both cell lines . E2 and ICI 182 , 780 down-regulated ERα expression . The number of U373 and D54 cells significantly increased after PPT ( ERα agonist ) treatment but not after DPN ( ERβ agonist ) one . To determine the role of Q15788 and Q9Y6Q9 coactivators in ERα induced cell growth , we silenced them with RNA interference . Coactivator silencing blocked the increase in cell number induced by PPT . The content of proteins involved in proliferation and metastasis was also determined after PPT treatment . Western blot analysis showed that in U373 cells the content of PR isoforms ( PR-A and PR-B ) , P00533 , P15692 and cyclin D1 increased after PPT treatment while in D54 cells only the content of P00533 was increased . Our results demonstrate that E2 induces cell growth of human astrocytoma cell lines through ERα and its interaction with Q15788 and Q9Y6Q9 and also suggest differential roles of ERα on cell growth depending on astrocytoma grade . 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 . 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 . In vivo activity of DB06080 , a multi-target kinase inhibitor , against acute myeloid leukemia with wild-type P36888 receptor . Neoangiogenesis plays an important role in leukemogenesis . We investigated the in vivo anti-leukemic effect of DB06080 against AML with wild-type P36888 using RFP transfected HL60 cells with in vivo imaging technology on both the subcutaneous and systemic leukemia xenograft models . DB06080 showed a five-fold inhibition of tumor growth in comparison with vehicle control . IHC analysis revealed that DB06080 decreased p- P17948 , Ki-67 labeling index , P15692 and remarkably increased apoptotic cells in the xenograft models . DB06080 also reduced the leukemia burden and prolonged survival . Our study supports the rationale for clinically testing an anti-angiogenesis agent in AML with wild-type P36888 . Molecular mechanisms involved in the growth stimulation of breast cancer cells by leptin . Obesity is a risk factor for breast cancer in postmenopausal women . Leptin , an adipocyte-derived cytokine , elicits proliferative effects in some cell types and potentially stimulates the growth of mammary epithelium . Here we show that leptin induced time- and dose-dependent signal transducer and activator of transcription 3 ( P40763 ) phosphorylation and extracellular signal-regulated kinase ( P29323 ) 1/2 kinase activation in breast carcinoma cells . Blocking P40763 phosphorylation with a specific inhibitor , AG490 , abolished leptin-induced proliferation of MCF-7 cells , whereas blocking P27361 /2 activation by a specific P27361 /2 kinase inhibitor , U0126 , did not result in any significant changes in leptin-induced cell proliferation . Our experiments also showed that one member of the P52701 family of steroid receptor coactivators , steroid receptor coactivator ( P12931 ) -1 , but not glucocorticoid receptor interacting protein 1 ( GRIP1 ) or amplified in breast cancer 1 ( Q9Y6Q9 ) , also functioned in gene transactivation in response to leptin treatment . O60760 pull-down experiments showed that Q15788 physically interacted with the activation domain of P40763 and that chromatin immunoprecipitation experiments detected the occupancy of Q15788 , but not GRIP1 or Q9Y6Q9 , on the promoter of P40763 target genes . Our experiments collectively showed that Q15788 is involved in P40763 signaling pathway that is implicated in leptin-stimulated cell growth . Selective interaction of vitamin D receptor with transcriptional coactivators by a vitamin D analog . The nuclear vitamin D receptor ( P11473 ) is a member of a nuclear receptor superfamily and acts as a ligand-dependent transcription factor . A family of cotranscriptional activators ( Q15788 , Q15596 , and Q9Y6Q9 ) interacts with and activates the transactivation function of nuclear receptors in a ligand-dependent way . We examined interaction of P11473 with these coactivators that was induced by several vitamin D analogs , since they exert differential subsets of the biological action of vitamin D through unknown mechanisms . Unlike other vitamin D analogs tested , O75051 ( 22-oxa-1alpha,25-dihydroxyvitamin D3 ) induced interaction of P11473 with Q15596 but not with Q15788 or Q9Y6Q9 . Consistent with these interactions , only Q15596 was able to potentiate the transactivation function of P11473 bound to O75051 . Thus , the present findings suggest that the structure of P11473 is altered in a vitamin D analog-specific way , resulting in selective interactions of P11473 with coactivators . Such selective interaction of coactivators with P11473 may specify the array of biological actions of a vitamin D analog like O75051 , possibly through activating a particular set of target gene promoters . 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 . 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 . HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing P16671 -dependent cholesteryl ester accumulation in macrophages . Protease inhibitors decrease the viral load in HIV patients , however the patients develop hypertriglyceridemia , hypercholesterolemia , and atherosclerosis . It has been assumed that protease inhibitor-dependent increases in atherosclerosis are secondary to the dyslipidemia . Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of P16671 and the accumulation of cholesteryl esters . The use of P16671 -blocking antibodies , a P16671 morpholino , and monocytes isolated from P16671 null mice demonstrated that protease inhibitor-induced increases in cholesteryl esters were dependent on P16671 upregulation . These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating P16671 and cholesteryl ester accumulation independent of dyslipidemia . Studies with P01130 null mice demonstrated that low doses of protease inhibitors induce an increase in the level of P16671 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids . Furthermore , the lack of P16671 protected the animals from protease inhibitor-induced atherosclerosis . Finally , ritonavir increased P37231 and P16671 mRNA levels in a PKC- and P37231 -dependent manner . We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of P16671 and the subsequent accumulation of sterol in macrophages . Permanent activation of P52926 in lipomas mimics its temporal physiological activation linked to the gain of adipose tissue . OBJECTIVE : In this study the activation of P52926 and overexpression by P05230 -driven stimulation of adipose tissue derived stem cells ( ADSCs ) in adipose tissue tumors were analyzed . In addition , the expression of P52926 and P37231 mRNA were quantified in canine subcutaneous abdominal adipose tissue from normal and overweight purebred dogs . DESIGN AND METHODS : ADSCs and adipose tissue explants stimulated with P05230 followed by gene expression analyses of P52926 and p14(Arf) using Western-blot and qRT-PCR . Furthermore , canine subcutaneous white adipose tissue ( WAT ) were analyzed by qRT-PCR for their expression of P52926 and P37231 . RESULTS : ADSCs and adipose tissue explants are able to execute a P52926 response upon P05230 stimulation . P05230 enhances proliferation of ADSCs by a P52926 -dependent mechanism . In lipomas increase of P52926 is accompanied by increased expression of p14(Arf) . Furthermore , a significantly elevated level of P52926 in overweight dogs and a negative correlation between the expression of P52926 and P37231 in subcutaneous cWAT were noted . CONCLUSIONS : These results suggest that WAT contains cells that as essential part of adipogenesis up-regulate P52926 resulting from growth factor stimulation . In subgroups of lipoma , constitutive activation of P52926 due to rearrangements replaces the temporal response triggered by growth factors . 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 . Inhibition of peroxisome proliferator-activated receptor gamma increases estrogen receptor-dependent tumor specification . P37231 ( PPARgamma ) is a nuclear receptor that regulates gene transcription associated with intermediary metabolism , adipocyte differentiation , and tumor suppression and proliferation . To understand the role of PPARgamma in tumorigenesis , transgenic mice were generated with mammary gland-directed expression of the dominant-negative transgene Pax8PPARgamma . Transgenic mice were phenotypically indistinguishable from wild-type ( WT ) mice , but mammary epithelial cells expressed a greater percentage of CD29(hi)/ P25063 (neg) , CK5(+) , and double-positive CK14/CK18 cells . These changes correlated with reduced P60484 and increased Ras and extracellular signal-regulated kinase ( P29323 ) and AKT activation . Although spontaneous tumorigenesis did not occur , transgenic animals were highly susceptible to progestin/7,12-dimethylbenz(a)anthracene-induced mammary carcinogenesis , which in contrast to WT mice resulted in a high tumor multiplicity and , most importantly , in the appearance of predominantly estrogen receptor alpha-positive ( ER(+) ) ductal adenocarcinomas . Tumors expressed a similar P60484 (lo)/pERK(hi)/pAKT(hi) phenotype as mammary epithelium and exhibited high activation of estrogen response element-dependent reporter gene activity . Tumorigenesis in MMTV-Pax8PPARgamma mice was insensitive to the chemopreventive effect of a PPARgamma agonist but was profoundly inhibited by the ER antagonist fulvestrant . These results reveal important new insights into the previously unrecognized role of PPARgamma in the specification of mammary lineage and the development of ER(+) tumors . 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) . DB01235 -treatment in primates disrupts the expression of A(2A) adenosine-CB(1) cannabinoid- P14416 heteromers in the caudate nucleus . The molecular basis of priming for DB01235 -induced dyskinesias in Parkinson 's disease ( PD ) , which depends on the indirect pathway of motor control , is not known . In rodents , the indirect pathway contains striatopallidal GABAergic neurons that express heterotrimers composed of A(2A) adenosine , CB(1) cannabinoid and D(2) dopamine receptors that regulate dopaminergic neurotransmission . The present study was designed to investigate the expression of these heteromers in the striatum of a primate model of Parkinson 's disease and to determine whether their expression and pharmacological properties are altered upon DB01235 treatment . By using the recently developed in situ proximity ligation assay and by identification of a biochemical fingerprint , we discovered a regional distribution of A(2A)/CB(1) /D(2) receptor heteromers that predicts differential D(2)-mediated neurotransmission in the caudate-putamen of Macaca fascicularis . Whereas heteromers were abundant in the caudate nucleus of both naïve and MPTP-treated monkeys , DB01235 treatment blunted the biochemical fingerprint and led to weak heteromer expression . These findings constitute the first evidence of altered receptor heteromer expression in pathological conditions and suggest that drugs targeting A(2A)-CB(1) -D(2) receptor heteromers may be successful to either normalize basal ganglia output or prevent DB01235 -induced side effects . 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 . 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 . 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 . A polymorphism associated with P40763 expression and response of chronic myeloid leukemia to interferon α . DB05258 ( IFN ) induces variable responses in chronic myeloid leukemia ( CML ) , with 8-30 % of early chronic phase cases achieving a complete cytogenetic response . We hypothesized that polymorphic differences in genes encoding IFN signal transduction components might account for different patient responses . We studied 174 IFN-treated patients , of whom 79 achieved less than 35 % Philadelphia-chromosome ( Ph ) positive metaphases ( responders ) and 95 failed to show any cytogenetic response ( more than 95 % Ph-positive metaphases ; non-responders ) . We compared 17 single nucleotide polymorphisms ( SNPs ) at P17181 , P48551 , P23458 , P29597 , P42224 , P40763 and STAT5a/b between the two groups and found a significant difference for rs6503691 , a SNP tightly linked to STAT5a , STAT5b and P40763 ( minor allele frequency 0.16 for non-responders ; 0.06 for responders , P=0.007 ) . Levels of P40763 mRNA correlated with rs6503691 genotype ( P < 0.001 ) as assessed by real time quantitative PCR and therefore we conclude that rs6503691 is associated with the P40763 expression levels and response of CML patients to IFN . A set of consensus mammalian mediator subunits identified by multidimensional protein identification technology . The Mediator is a multiprotein transcriptional coactivator that is expressed ubiquitously in eukaryotes from yeast to mammals and is required for induction of RNA polymerase II ( pol II ) transcription by DNA binding transcription factors . In the work described here , we exploit multidimensional protein identification technology ( MudPIT ) to carry out a proteomic analysis of the subunit composition of the mammalian Mediator complex . By comparing MudPIT data sets obtained from six independent Mediator preparations immunoaffinity purified through their Nut2 ( Q9BTT4 ) , Med25 ( Q9NWA0 ) , Intersex ( Q9NX70 ) , A0JLT2 ( A0JLT2 ) , AK007855 ( Q9H204 ) , or CRSP70 ( O95402 ) subunits , we identify a set of consensus mammalian Mediator subunits . In addition , we identify as Mediator-associated proteins the P49336 -like cyclin-dependent kinase CDK11 and the Q9UHV7 -like Q71F56 protein ( Q71F56 ) , which is mutated in patients with the congenital heart defect transposition of the great arteries ( TGA ) . 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 .

MH_train_1414

MH_train_1414

MH_train_1414

interacts_with DB00741?

multiple_choice

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 . 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 . 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 . Inhibitors of arachidonic acid metabolism reduce DNA and nuclear fragmentation induced by P01375 plus cycloheximide in U937 cells . U937 human myeloid leukemia cells are induced to apoptosis by tumour necrosis factor ( P01375 ) plus cycloheximide ( CHX ) . We have analysed the effect of various inhibitors of the arachidonic acid ( AA ) metabolism on several features of this process . The formation of high molecular weight and oligonucleosomal DNA fragments as well as nuclear fragmentation were reduced by inhibitors of P09917 ( BWA4C and BWB70C ) , P09917 activating protein ( MK-886 ) , and cytosolic P04054 ( AACOCF3 ) . None of these agents blocked the morphological changes detected by microscopy or flow cytometry , phosphatidylserine exposure on the cell surface or Caspase 3-like activation . AA also induced nuclear fragmentation at a concentration of 1-20 microM . However , the mechanisms by which these inhibitors act , remain unexplained since there was no P09917 expression in the U937 cells and no AA release followed their stimulation with P01375 plus CHX . 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 . 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 . 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 . 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 . Influence of labor on neonatal neutrophil apoptosis , and inflammatory activity . Neutrophil apoptosis is impaired in neonates , and this contributes to prolonged inflammation and tissue injury in infants after infection or trauma . In the present studies , we investigated whether labor generates mediators that further suppress apoptosis . We found that neutrophil apoptosis was reduced in neonates exposed to labor , when compared with infants delivered by cesarean section before labor . This was not due to alterations in caspase-3 or inhibitor of apoptosis protein-2 ( IAP-2 ) . In contrast , labor primed neutrophils to express tumor necrosis factor alpha ( P01375 ) , suggesting that proinflammatory mediators contribute to reduced apoptosis after labor . Eicosanoids generated via cyclooxygenase-2 ( Cox-2 ) and lipoxygenase ( Lox ) also regulate neutrophil apoptosis . 15-Lox , which generates proapoptotic lipoxins , but not Cox-2 , was greater in neutrophils before labor , relative to cells exposed to labor . Anti-inflammatory eicosanoids exert their effects in part via peroxisome proliferator-activated receptor gamma ( P37231 ) . Expression of gelatinase-associated lipocalin and catalase , two markers of P37231 activity , were increased in neonatal neutrophils before labor , relative to cells exposed to labor . These findings suggest that the anti-inflammatory environment is maintained before labor , in part , by eicosanoids . Although increased neutrophil longevity after labor is important for host defense in the immediate newborn period , it may contribute to inflammatory or oxidative injury in susceptible infants . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . 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 . Phosphodiesterase 4 inhibitors reduce human dendritic cell inflammatory cytokine production and Th1-polarizing capacity . Inhibitors of DB02527 -specific phosphodiesterase ( PDE ) 4 have been shown to inhibit inflammatory mediator release and T cell proliferation , and are considered candidate therapies for T(h)1-mediated diseases . However , little is known about how DB05876 inhibitors influence dendritic cells ( DC ) , the cells responsible for the priming of naive T(h) cells . Therefore , we investigated the PDE profile of monocyte-derived DC , and whether DB05876 inhibitors modulate DC cytokine production and T cell-polarizing capacity . We mainly found DB02527 -specific DB05876 enzymatic activity in both immature and mature DC . In contrast to monocytes that mainly express Q07343 , we found that P27815 is the predominant DB05876 subtype present in DC . Immature DC showed reduced ability to produce IL-12p70 and tumor necrosis factor ( P01375 ) -alpha upon lipopolysaccharide or P29965 ( P29965 ) stimulation in the presence of DB05876 inhibitors , whereas cytokine production upon P29965 stimulation of fully mature DC in the presence of DB05876 inhibitors was not affected . Exposure to DB05876 inhibitors for 2 days during DC maturation did not influence T cell-stimulatory capacity or acquisition of a mature phenotype , but increased the expression of the chemokine receptor P61073 . Furthermore , DC matured in the presence of DB05876 inhibitors showed reduced capacity to produce IL-12p70 and P01375 upon subsequent P29965 stimulation . Using these DB05876 inhibitor-matured DC to stimulate naive T cells resulted in a reduction of P01579 -producing ( T(h)1 ) cells . These findings indicate that DB05876 inhibitors can affect T cell responses by acting at the DC level and may increase our understanding of the therapeutic implication of DB05876 inhibitors for T(h)1-mediated disorders . Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors . The human HDAC ( histone deacetylase ) family , a well-validated anticancer target , plays a key role in the control of gene expression through regulation of transcription . While HDACs can be subdivided into three main classes , the class I , class II and class III HDACs ( sirtuins ) , it is presently unclear whether inhibiting multiple HDACs using pan-HDAC inhibitors , or targeting specific isoforms that show aberrant levels in tumours , will prove more effective as an anticancer strategy in the clinic . To address the above issues , we have tested a number of clinically relevant HDACis ( HDAC inhibitors ) against a panel of rhHDAC ( recombinant human HDAC ) isoforms . Eight rhHDACs were expressed using a baculoviral system , and a Fluor de Lystrade mark ( Biomol International ) HDAC assay was optimized for each purified isoform . The potency and selectivity of ten HDACs on class I isoforms ( rhHDAC1 , rhHDAC2 , rhHDAC3 and rhHDAC8 ) and class II HDAC isoforms ( rhHDAC4 , rhHDAC6 , rhHDAC7 and rhHDAC9 ) was determined . MS-275 was Q13547 -selective , DB05651 was Q13547 - and Q92769 -selective , apicidin was Q92769 - and O15379 -selective and valproic acid was a specific inhibitor of class I HDACs . The hydroxamic acid-derived compounds ( trichostatin A , DB00238 -LAQ824 , DB06603 , ITF2357 , vorinostat and belinostat ) were potent pan-HDAC inhibitors . The growth-inhibitory effect of the HDACis on HeLa cells showed that both pan-HDAC and class-I-specific inhibitors inhibited cell growth . The results also showed that both pan-HDAC and class-I-specific inhibitor treatment resulted in increased acetylation of histones , but only pan-HDAC inhibitor treatment resulted in increased tubulin acetylation , which is in agreement with their activity towards the Q9UBN7 isoform . Variations of P60568 , P05231 , P01375 alpha plasmatic levels in relapsing remitting multiple sclerosis . We performed a longitudinal analysis of serum P60568 , P05231 and P01375 alpha concentrations in 40 relapsing remitting MS patients and 20 healthy subjects . Disease activity was quantified by Minimal Record of Disease ( M.R.D. ) for MS , every 2 or 3 months . P60568 , P05231 P01375 alpha production was analysed without and with PHA stimulation of whole blood for 2 hours at 37 degrees C . No significant change in P60568 level was found in MS serum . Individual P01375 alpha production was significantly increased ( P < 0.007 ) during relapses . The global spontaneous P05231 production was markedly higher in the relapse group than in the control group ( P < 0.01 ) and than in the remission group ( P < 0.002 ) without significant individual variations of cytokine levels regarding the disease activity . Productions of cytokines were enhanced by PHA stimulation , a condition that however suppressed the differences observed without mitogen stimulation . Our data suggest that P01375 alpha could be a marker for relapses while P05231 might reflect the global activity of the immune system in MS . c-Jun N-terminal kinase and Akt signalling pathways regulating tumour necrosis factor-α-induced interleukin-32 expression in human lung fibroblasts : implications in airway inflammation . Airway inflammatory diseases such as chronic obstructive pulmonary disease ( P48444 ) and asthma are associated with elevated expression of interleukin-32 ( P24001 ) , a recently described cytokine that appears to play a critical role in inflammation . However , so far , the regulation of pulmonary P24001 production has not been fully established . We examined the expression of P24001 by tumour necrosis factor-α ( P01375 -α ) in primary human lung fibroblasts . Human lung fibroblasts were cultured in the presence or absence of P01375 -α and/or other cytokines/Toll-like receptor ( TLR ) ligands or various signalling molecule inhibitors to analyse the expression of P24001 by quantitative RT-PCR and ELISA . Next , activation of Akt and c-Jun N-terminal kinase ( JNK ) signalling pathways was investigated by Western blot . P24001 mRNA of four spliced isoforms ( α , β , γ and δ ) was up-regulated upon P01375 -α stimulation , which was associated with a significant P24001 protein release from P01375 -α-activated human lung fibroblasts . The combination of interferon-γ and P01375 -α induced enhanced P24001 release in human lung fibroblasts , whereas P05112 , Q16552 , Q8TAD2 and TLR ligands did not alter P24001 release in human lung fibroblasts either alone , or in combination with P01375 -α . Furthermore , the activation of Akt and JNK pathways regulated P01375 -α-induced P24001 expression in human lung fibroblasts , and inhibition of the Akt and JNK pathways was able to suppress the increased release of P24001 to nearly the basal level . These data suggest that P01375 -α may be involved in airway inflammation via the induction of P24001 by activating Akt and JNK signalling pathways . Therefore , the P01375 -α/ P24001 axis may be a potential therapeutic target for airway inflammatory diseases . 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 . 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 . Essential oil of niaouli preferentially potentiates antigen-specific cellular immunity and cytokine production by macrophages . In vivo immunomodulatory effect of essential oil of niaouli ( EON ) was investigated using a mouse model , in which mice were immunized with DB05299 ( KLH ) and intraperitoneally given EON ( less than 500 microl kg(-1) body weight ) . In vivo efficacy of EON for immune potentiation was convinced by significantly higher expression of an activation marker , CD25 , on freshly isolated draining lymph node ( LN ) T cells , but not B cells . However , immunofluoresence analysis failed to show any proportional change in T/B and P01730 (+)/CD8(+) T cell ratios . Data of KLH-specific immunoglobulin serum levels showed that EON does not affect humoral immune response . Instead , proliferative response and IFNgamma production of LN T cells ex vivo stimulated with KLH were significantly higher in EON-treated group , but not P60568 and P05112 production . These results clearly show that EON preferentially upregulates T-cell mediated cellular immunity . We further clarified the accessory cells ' contribution to the EON-mediated potentiation of cellular immunity and found considerably higher production of and P01375 and IL-12 by splenic macrophages from EON-treated mice when stimulated with lipopolysaccharide ( LPS ) and IFNgamma . Collectively , in vivo EON treatment potentiates T cell-mediated cellular immunity and macrophage activity , but not humoral immunity . The current study provides a rationale for clinical application of EON to control infectious diseases , in particular , those caused by intracellular pathogens .

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interacts_with DB01356?

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Alfa and beta estrogen receptors and the biliary tree . This manuscript summarizes recent data showing that estrogens and their receptors play an important role in modulating cholangiocyte proliferation . We have recently demonstrated that rat cholangiocytes express both estrogen receptors ( ER ) -alpha and -beta subtypes , while hepatocytes only express P03372 . ER and especially the Q92731 subtype , are overexpressed in cholangiocytes proliferating after bile duct ligation ( BDL ) in the rat , in association with enlarged bile duct mass and with enhanced estradiol serum levels . Cholangiocyte proliferation , during BDL , is impaired by estrogen antagonists ( tamoxifen , DB00947 ) which furthermore , induce the overexpression of Fas antigen and activate apoptosis of proliferating cholangiocytes . 17beta-estradiol stimulates , in vitro cholangiocyte proliferation , and this effect is individually blocked by tamoxifen or DB00947 . Cholangiocyte proliferation during BDL was associated with an enhanced protein expression of phosphorylated extracellular regulated kinases ( P29323 )1/2 which is , in contrast , negatively modulated by tamoxifen in association with its antiproliferative effect . This indicates a major involvement of the P29323 system in the estrogen modulation of cholangiocyte proliferation . 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 . 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 . DB09341 - and interleukin-1beta-induced beta-cell apoptosis requires Ca2+ influx and extracellular signal-regulated kinase ( P29323 ) 1/2 activation and is prevented by a sulfonylurea receptor 1/inwardly rectifying K+ channel 6.2 ( Q09428 /Kir6.2 ) selective potassium channel opener in human islets . Increasing evidence indicates that a progressive decrease in the functional beta-cell mass is the hallmark of both type 1 and type 2 diabetes . The underlying causes , beta-cell apoptosis and impaired secretory function , seem to be partly mediated by macrophage production of interleukin ( IL ) -1beta and/or high-glucose-induced beta-cell production of IL-1beta . Treatment of type 1 and type 2 diabetic patients with the potassium channel opener diazoxide partially restores insulin secretion . Therefore , we studied the effect of diazoxide and of the novel potassium channel opener NN414 , selective for the beta-cell potassium channel Q09428 /Kir6.2 , on glucose- and IL-1beta-induced apoptosis and impaired function in human beta-cells . Exposure of human islets for 4 days to 11.1 and 33.3 mmol/l glucose , 2 ng/ml IL-1beta , or 10 and 100 micromol/l of the sulfonylurea tolbutamide induced beta-cell apoptosis and impaired glucose-stimulated insulin secretion . The deleterious effects of glucose and IL-1beta were blocked by 200 micromol/l diazoxide as well as by 3 and 30 micromol/l NN414 . By Western blotting with phosphospecific antibodies , glucose and IL-1beta were shown to activate the extracellular signal-regulated kinase ( P29323 ) 1/2 , an effect that was abrogated by 3 micromol/l NN414 . Similarly , 1 micromol/l of the mitogen-activated protein kinase/ P29323 kinase 1/2 inhibitor PD098059 or 1 micromol/l of the l-type Ca(2+) channel blocker nimodipine prevented glucose- and IL-1beta-induced P29323 activation , beta-cell apoptosis , and impaired function . Finally , islet release of IL-1beta in response to high glucose could be abrogated by nimodipine , NN414 , or PD098059 . Thus , in human islets , glucose- and IL-1beta-induced beta-cell secretory dysfunction and apoptosis are Ca(2+) influx and P29323 dependent and can be prevented by the beta-cell selective potassium channel opener NN414 . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . The BH3 mimetic DB05764 synergizes with the Q02750 /2 inhibitor selumetinib/AZD6244 to promote O43521 -dependent tumour cell death and inhibit acquired resistance . Tumour cells typically exhibit a G(1) cell cycle arrest in response to the Q02750 /2 [ mitogen-activated protein kinase/ P29323 ( extracellular-signal-regulated kinase ) kinase 1/2 ] inhibitor selumetinib , but do not die , and thus they acquire resistance . In the present study we examined the effect of combining selumetinib with the BH3 [ P10415 ( B-cell lymphoma 2 ) homology domain 3 ] -mimetic P10415 inhibitor DB05764 . Although either drug alone caused little tumour cell death , the two agents combined to cause substantial caspase-dependent cell death and inhibit long-term clonogenic survival of colorectal cancer and melanoma cell lines with P15056 (V600E) or DB01367 mutations . This cell death absolutely required Q07812 ( P10415 -associated X protein ) and was inhibited by RNAi ( RNA interference ) -mediated knockdown of O43521 ( P10415 -interacting mediator of cell death ) in the P15056 (V600E)-positive COLO205 cell line . When colorectal cancer cell lines were treated with selumetinib plus DB05764 we observed a striking reduction in the incidence of cells emerging with acquired resistance to selumetinib . Similar results were observed when we combined DB05764 with the P15056 (V600E)-selective inhibitor PLX4720 , but only in cells expressing P15056 (V600E) . Finally , cancer cells in which acquired resistance to selumetinib arises through P15056 (V600E) amplification remained sensitive to DB05764 , whereas selumetinib-resistant HCT116 cells ( P01116 (G13D) amplification ) were cross-resistant to DB05764 . Thus the combination of a P10415 inhibitor and an P27361 /2 pathway inhibitor is synthetic lethal in P27361 /2-addicted tumour cells , delays the onset of acquired resistance and in some cases overcomes acquired resistance to selumetinib . Heme oxygenase-1 attenuates glucose-mediated cell growth arrest and apoptosis in human microvessel endothelial cells . Heme oxygenase-1 ( P09601 ) is a stress protein that has been suggested to participate in defense mechanisms against agents that may induce oxidative injury , such as heme and inflammatory molecules . Incubation of endothelial cells in a high-glucose ( 33 mmol/L ) medium for 7 days resulted in a decrease of HO activity by 34 % and a decrease in P09601 and P30519 proteins compared with cells exposed to low glucose ( 5 mmol/L ) ( P < 0.05 ) or cells exposed to mannitol ( 33 mmol/L ) . Overexpression of P09601 was coupled with an increase in HO activity and carbon monoxide synthesis , decreased cellular heme , and acceleration in all phases of the cell cycle ( P < 0.001 ) . The rate of cell cycle or cell birth rate was increased by 29 % ( P < 0.05 ) in cells overexpressing P09601 but decreased by 23 % ( P < 0.05 ) in cells underexpressing P09601 compared with control cells . Exposure to high glucose significantly decreased cell-cycle progression in control cells and in cells underexpressing P09601 but did not decrease cell-cycle progression in cells overexpressing P09601 . High glucose induced P38936 and p27 in control cells but not in cells overexpressing P09601 . The addition of DB04912 ( DB04912 ) , an inhibitor of HO activity , reversed the P09601 -mediated decrease of P38936 and p27 in cells overexpressing P09601 . These findings identify a novel effect of P09601 on endothelial cell growth and indicate that heme metabolism and P09601 expression regulate signaling systems in cells exposed to high glucose , which controls cell-cycle progression . DB03203 1-phosphate-induced cell proliferation , survival , and related signaling events mediated by G protein-coupled receptors Edg3 and Edg5 . DB03203 1-phosphate ( Q14703 ) regulates cell proliferation , apoptosis , motility , and neurite retraction . Contradictory reports propose that Q14703 acts as either an intracellular second messenger or an extracellular ligand for cell-surface receptors . Hence , the precise signaling mechanisms mediating the diverse cellular effects of Q14703 remain to be determined . Here , we investigate whether Q14703 stimulation of cell proliferation , survival , and related signaling events can be mediated by the recently cloned Edg family members of G protein-coupled receptors . We observed that Q14703 treatment significantly increased proliferation of HTC4 hepatoma cells stably transfected with human Q14703 receptor Edg3 or Edg5 , which was attributable to stimulation of cell growth and inhibition of apoptosis caused by serum starvation . Edg3 and Edg5 transduced Q14703 -evoked signaling events relevant to cell proliferation and survival , including activation of the P29323 / Q96HU1 kinases , and immediate-early induction of c-Jun and c-Fos . Trancriptional activation of reporter genes for the c-fos promoter and the serum response element by Edg3 and Edg5 transfected in Jurkat cells was inhibited by pertussis toxin and P01024 exoenzyme , implicating G(i/o)- and Rho-dependent pathways . Our data also indicated that Edg3 and Edg5 mediated the serum response element activation through transcriptional factors Elk-1 and serum response factor . Thus , specific G protein-coupled receptors Edg3 and Edg5 account for , at least in part , Q14703 -induced cell proliferation , survival , and related signaling events . Growth hormone releasing hormone induces the expression of nitric oxide synthase . Growth hormone releasing hormone ( P01286 ) and its receptors are expressed in a wide variety of human tumours and established cancer cell lines and are involved in carcinogenesis . In addition , P01286 antagonists exert an antitumour activity in experimental cancer models . Recent studies indicate that the mechanisms involved in the mediation of the effects of P01286 include the regulation of the metabolism of the reactive oxygen species . This work demonstrates the expression of P01286 receptors and P01286 in the A549 human lung cancer cell line and shows that the mitogenic effect of P01286 in these cells is dependent on the activation of the extracellular receptor kinase ( P29323 )1/2 pathway . The action of P01286 can be suppressed by P01286 antagonist MZ-5-156 and mitogen activated protein kinase ( MAPK ) inhibitor PD 098059 . These results are reflected in the effect in the proliferating cell nuclear antigen . In addition , our study shows that P01286 increases the expression of the inducible nitric oxide synthase , an enzyme which is strongly involved in various human diseases , including cancer and augments key intracellular regulators of its expression , such as pNF ( nuclear factor ) κBp50 and cyclooxygenase 2 . P01286 antagonist MZ-5-156 counteracts the effects of P01286 in these studies , indicating that this class of peptide antagonists may be useful for the treatment of diseases related to increased oxidative and nitrosative stress . [ P08709 -- new physiopathological and therapeutic aspects ] . Factor VII ( FVII ) is a vitamin K dependent coagulation factor that is synthesized in the liver , where clearance of the unactivated protein also takes place . It is a glycoprotein that following activation plays an important role in initiating coagulation after complex formation with tissue factor . A revised hypothesis of blood coagulation implicating the requirement of intact extrinsic and intrinsic pathways is presented . Increased activity of factor VII is associated with atherogenesis , and FVII deficiency may cause bleeding disorders . Recombinant FVIIa ( DB00036 ) may be used in the treatment of haemophilic patients with antibodies against factor VIII or factor IX , utilizing its direct action in activation of factor X . Ongoing studies are investigating whether DB00036 can shorten the bleeding time in patients with thrombocytopenia . The effects of mitiglinide ( KAD-1229 ) , a new anti-diabetic drug , on DB00171 -sensitive K+ channels and insulin secretion : comparison with the sulfonylureas and nateglinide . DB01252 ( KAD-1229 ) , a new anti-diabetic drug , is thought to stimulate insulin secretion by closing the DB00171 -sensitive K+ ( K( DB00171 ) ) channels in pancreatic beta-cells . However , its selectivity for the various K( DB00171 ) channels is not known . In this study , we examined the effects of mitiglinide on various cloned K( DB00171 ) channels ( Kir6.2/ Q09428 , Kir6.2/SUR2A , and Kir6.2/SUR2B ) reconstituted in COS-1 cells , and compared them to another meglitinide-related compound , nateglinide . Patch-clamp analysis using inside-out recording configuration showed that mitiglinide inhibits the Kir6.2/ Q09428 channel currents in a dose-dependent manner ( IC50 value , 100 nM ) but does not significantly inhibit either Kir6.2/SUR2A or Kir6.2/SUR2B channel currents even at high doses ( more than 10 microM ) . DB00731 inhibits Kir6.2/ Q09428 and Kir6.2/SUR2B channels at 100 nM , and inhibits Kir6.2/SUR2A channels at high concentrations ( 1 microM ) . Binding experiments on mitiglinide , nateglinide , and repaglinide to Q09428 expressed in COS-1 cells revealed that they inhibit the binding of [3H]glibenclamide to Q09428 ( IC50 values : mitiglinide , 280 nM ; nateglinide , 8 microM ; repaglinide , 1.6 microM ) , suggesting that they all share a glibenclamide binding site . The insulin responses to glucose , mitiglinide , tolbutamide , and glibenclamide in MIN6 cells after chronic mitiglinide , nateglinide , or repaglinide treatment were comparable to those after chronic tolbutamide and glibenclamide treatment . These results indicate that , similar to the sulfonylureas , mitiglinide is highly specific to the Kir6.2/ Q09428 complex , i.e. , the pancreatic beta-cell K( DB00171 ) channel , and suggest that mitiglinide may be a clinically useful anti-diabetic drug . 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 . DB03203 1-phosphate inhibits nitric oxide production induced by interleukin-1beta in rat vascular smooth muscle cells . DB03203 1-phosphate ( Q14703 ) is a lipid mediator that exerts potent and diverse biological effects on several cardiovascular cells . We investigated the effect of Q14703 on interleukin ( IL ) -1beta-induced nitric oxide ( NO ) production and inducible NO synthase ( P35228 ) expression in rat vascular smooth muscle cells ( VSMCs ) . Q14703 inhibited NO production at concentrations higher than 0.1 muM ; this was associated with the inhibition of P35228 protein and mRNA expression . Q14703 also inhibited IL-1beta-induced P30793 ( GTPCH ) mRNA expression . Pertussis toxin ( PTX ) partially attenuated the inhibitory effects of Q14703 on NO production and P35228 protein induction , whereas it completely blocked the inhibitory effects on P35228 and GTPCH mRNA expression . Q14703 inhibited P35228 expression in Ca(2+)-depleted conditions ; PTX did not modify this effect . The Rho kinase inhibitor Y 27632 partially but significantly attenuated the inhibitory effect of Q14703 on P35228 expression in Ca(2+)-depleted condition but did not affect it in the presence of Ca(2+) . Q14703 significantly inhibited IL-1beta-induced persistent activation of extracellular signal-regulated kinase ( P29323 ) but had no effect in Ca(2+)-depleted conditions . Thus , Q14703 inhibits IL-1beta induction of NO production and P35228 expression in rat VSMCs through multiple mechanisms involving both PTX-sensitive and -insensitive G proteins coupled to Q14703 receptors . Furthermore , Ca(2+)-dependent P29323 inhibition and Ca(2+)-independent Rho kinase activation might be involved in the inhibitory mechanism of P35228 expression . Through its action on NO production by VSMCs , Q14703 may play an important role in the progression of local vascular injury associated with thrombosis , atherosclerosis , and hypertension . Repression of human heat shock factor 1 activity at control temperature by phosphorylation . Human heat shock transcription factor 1 ( Q00613 ) is responsible for stress-induced transcription of heat shock protein genes . The activity of the Q00613 transcriptional activation domains is modulated by a separate regulatory domain , which confers repression at control temperature and heat inducibility . We show here that two specific proline-directed serine motifs are important for function of the regulatory domain : Mutation of these serines to alanine derepresses Q00613 activity at control temperature , and mutation to glutamic acid , mimicking a phosphorylated serine , results in normal repression at control temperature and normal heat shock inducibility . Tryptic mapping shows that these serines are the major phosphorylation sites of Q00613 at control temperature in vivo . Stimulation of the Raf/ P29323 pathway in vivo results in an increased level of phosphorylation of these major sites and the regulatory domain is an excellent substrate in vitro for the mitogen-activated MAPK/ P29323 . We conclude that phosphorylation of the regulatory domain of Q00613 decreases the activity of Q00613 at control temperature , and propose a mechanism for modification of Q00613 activity by growth control signals . Oligodendroglia are protected from antibody-mediated complement injury by normal immunoglobulins ( " DB00028 " ) . High-dose intravenous immunoglobulin ( DB00028 ) treatment has become a promising immune therapy that can modulate the immune system at several levels , including the complement cascade . In relation to inflammatory demyelinating disease , there is some clinical evidence for the suppression of disease activity by DB00028 , while a role in promoting remyelination after experimental myelin damage has been described . Antibody and complement deposition have been implicated in the immune attack in some cases of multiple sclerosis ( MS ) , and to investigate the mechanisms of action of DB00028 , we studied the effect of DB00028 using the model of complement-mediated cell injury on oligodendroglia in vitro . There was no effect on direct complement lysis of the oligodendroglial cell line CG4 , but antibody-dependent complement damage was inhibited in a dose-dependent manner by DB00028 . These results were confirmed with primary cultures of oligodendrocyte precursor cells ( OPC ) and oligodendrocytes . The addition of excess C1 , P01024 , and C4 did not influence the inhibitory effect of DB00028 , implying that binding of these complement components does not play a role , in contrast to other experimental models of complement damage . F(ab')2 immunoglobulin fragments were at least partially responsible for the effect . We conclude that DB00028 may be protective in antibody-mediated complement injury of oligodendrocytes and their progenitors , and that this effect is likely to be mediated via antibody binding , rather than interference with complement activation . Inhibition of inflammatory mechanisms , as opposed to a direct effect on remyelinating cells , may underlie the role of DB00028 in promoting myelin repair in experimental models . Overexpression of nuclear distribution protein ( hNUDC ) causes pro-apoptosis and differentiation in Dami megakaryocytes . OBJECTIVES : Overexpression of hNUDC , a member of the nuclear distribution protein family , reduces cell population growth in prostate cancer cell lines , concurrent with induced morphological change and enhanced polyploidization . These phenomena are also closely associated with terminal phases of megakaryocyte maturation . MATERIALS AND METHODS : In Dami cells , MTT and trypan blue assays were used to investigate cell viability and proliferation effects of hNUDC , and flow cytometry was used to analyse cell cycle and DNA content . Real-time RT-PCR was employed to detect mRNA expression . Activations of caspase-3 , P29323 , Akt and Stat-5 were determined by immunoblotting . May-Grünwald-Giemsa staining was performed to reveal cell morphology . RESULTS AND CONCLUSION : Functional studies using adenovirus-mediated hNUDC overexpression led to inhibition of megakaryocyte proliferation via cell cycle arrest in G2/M transition phase . This process could have been be mediated by upregulation of P38936 and downregulation of its downstream targets , including cyclin B1 , cyclin B2 and c-myc . Enhanced apoptosis in turn ensued , characterized by increased caspase-3 activation , upregulation of pro-apoptotic Bax and downregulation of anti-apoptotic Bcl-2 . Furthermore , hNUDC overexpression elevated the level of megakaryocyte maturation , associated with increased polyploidy , cell morphological changes and increased expression of cell surface differentiation markers , including CD10 , P16070 , CD41 and CD61 . Our results further suggest that the P29323 signalling pathway was involved in hNUDC overexpression-induced apoptosis . Taken together , this study provides experimental evidence for overexpression of hNUDC in Dami cells and suggests that activation of apoptotic machinery may be involved in megakaryocytic differentiation . DB02424 attenuates 3‑nitropropionic acid‑induced apoptosis and JNK activation through the expression of HSP 70 in striatal cells . Although selective striatal cell death is a characteristic hallmark in the pathogenesis of Huntington 's disease ( HD ) , the underlying mechanism of striatal susceptibility remains to be clarified . Heat shock proteins ( HSPs ) have been reported to suppress the aggregate formation of mutant huntingtin and concurrent striatal cell death . In a previous study , we observed that heat shock transcription factor 1 ( Q00613 ) , a major transcription factor of HSPs , significantly attenuated 3‑nitropropionic acid (3NP)‑induced reactive oxygen species ( ROS ) production and apoptosis through the expression of HSP 70 in striatal cells . To investigate the differential roles of HSPs in 3NP‑induced striatal cell death , the effect of geldanamycin ( GA ) , an P08238 inhibitor , was examined in 3NP‑stimulated striatal cells . GA significantly attenuated 3NP‑induced striatal apoptosis and ROS production with an increased expression of HSP 70 . Triptolide ( TL ) , an HSP 70 inhibitor , abolished GA‑mediated protective effects in 3NP‑stimulated striatal cells . To understand the underlying mechanism by which GA‑mediated HSP 70 protects striatal cells against 3NP stimulation , the involvement of various signaling pathways was examined . GA significantly attenuated 3NP‑induced c‑Jun N‑terminal kinase ( JNK ) phosphorylation and subsequent c‑Jun phosphorylation in striatal cells . Taken together , the present study demonstrated that GA exhibits protective properties against 3NP‑induced apoptosis and JNK activation via the induction of HSP 70 in striatal cells , suggesting that expression of HSP 70 may be a valuable therapeutic target in the treatment of HD .

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interacts_with DB00834?

multiple_choice

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 . Downregulation of class II transactivator ( P33076 ) expression by synthetic cannabinoid CP55,940 . Cannabinoid receptors are known to be expressed in microglia ; however , their involvement in specific aspects of microglial immune function has not been demonstrated . Many effects of cannabinoids are mediated by two G-protein coupled receptors , designated P21554 and CB2 . We have shown that the P21554 receptor is expressed in microglia that also express MHC class II antigen ( J. Neuroimmunol. 82 ( 1998 ) 13-21 ) . In our present study , we have analyzed the effect of cannabinoid agonist CP55,940 on MHC class II expression on the surface of P01579 induced microglial cells by flow cytometry . CP55,940 blocked the class II MHC expression induced by P01579 . It has been shown that the regulation of class II MHC genes occurs primarily at the transcriptional level , and a non-DNA binding protein , class II transactivator ( P33076 ) , has been shown to be the master activator for class II transcription . We find that mRNA levels of P33076 are increased in P01579 induced EOC 20 microglial cells and that this increase is almost entirely eliminated by the cannabinoid agonist CP55,940 . These data suggests that cannabinoids affect MHC class II expression through actions on P33076 at the transcriptional level . 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 . 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 . The bile acid receptor Q96RI1 is a modulator of intestinal innate immunity . The farnesoid X receptor ( Q96RI1 ) is a bile acid-regulated nuclear receptor expressed in enterohepatic tissues . In this study we investigated whether Q96RI1 is expressed by cells of innate immunity and regulates inflammation in animal models of colitis . Acute ( 7 days ) and chronic ( 8 wk ) colitis were induced in wild-type and Q96RI1 (-/-) mice by intrarectal administration of trinitrobenzensulfonic acid or by 7-day administration of 5 % dextran sulfate in drinking water . The results of this experiment demonstrate that Q96RI1 is expressed by and exerts counterregulatory effects on cells of innate immunity . Exposure of LPS-activated macrophages to 6-ethyl chenodeoxycholic acid ( 6E- DB06777 ; INT-747 ) a synthetic Q96RI1 ligand , results in a reciprocal regulation of NF-kappaB dependent-genes ( P01375 , IL-1beta , P05231 , P23219 , P35354 , and P35228 ) and induction of Q15466 , a Q96RI1 -regulated gene . Q96RI1 activation stabilizes the nuclear corepressor NCoR on the NF-kappaB responsive element on the IL-1beta promoter . Colon inflammation in Crohn 's disease patients and in rodent models of colitis is associated with a reduced expression of Q96RI1 mRNA . Using two rodent models of colon inflammation , we show that progression of these immune-mediated disorders is exacerbated in Q96RI1 (-/-) mice ( p < 0.01 ) . In vivo treatment with INT-747 attenuates organ injury and immune cell activation . Q96RI1 activation increased the colon expression of P51161 , Q96RI1 , and Q15466 while reducing IL-1beta , P60568 , P05231 , P01375 , and P01579 mRNA expression and attenuating disease severity . In aggregate , these findings provide evidence that Q96RI1 is an essential component of a network of nuclear receptors that regulate intestinal innate immunity and homeostasis . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . 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 . Role of DB01277 receptor in radiation response . P08069 ( IGF-1R ) is a transmembrane receptor tyrosine kinase involved in the development and progression of cancer whose activation strongly promotes cell growth and survival . IGF-1R exerts its main actions through the activation of the mitogen-activated protein kinase and phosphoinositide 3-kinase pathways . In addition to their traditional roles , IGF-1R activation has been associated with increased radioresistance both in vitro and in vivo , although the molecular mechanisms behind this process are still unclear . Recently , IGF-1R has been associated to new partners as major vault proteins , BCL-2 , Q07812 , or P12956 /80 , related to radiochemotherapy resistance , regulation of apoptosis , and nonhomologous end-joining DNA repair . Here , we review these novel associations of IGF-1R trying to explain the resistance to radiotherapy mediated by IGF-1R . Finally , we revised the role of new therapies leading to block the receptor to enhance the efficacy of radiation . 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 . A new rabbit model for the study on cervical compressive myelopathy . Development process and pathology of myelopathy due to chronic spinal cord compression have not been fully elucidated . This study was conducted in order to establish an experimental model which can efficiently produce myelopathy and be useful in the studies on myelopathy due to chronic spinal cord compression . Under electrophysiological monitoring of the spinal cord , anterior compression was produced on P01031 using a plastic screw . Two weeks later , a plastic plate was inserted under the P01031 arch . For the subsequent 10 months on average , walking pattern and MR images were periodically monitored . Before the sacrifice , electrophysiological test was performed and then histopathological examination was done . Palsy appeared at 5 months on average after the addition of posterior compression . Mean compression ratio of the spinal cord calculated on MR images was 34 % . All animals with compression showed a high intramedullary signal intensity , and the mean contrast-to-noise ratio ( P21554 ) in the compressed area was 49 % . Electrophysiological test showed a significant decrease in the amplitude of spinal cord evoked potentials ( SCEPs ) at the given compression level . Histology showed flattening of the anterior horn , disappearance and necrosis of anterior horn cells in the gray matter ; and demyelination and axonal degeneration in the white matter . The antero-posterior compression produces the condition of spinal canal stenosis . Repeated antero-posterior compression to the spinal cord is important in establishing myelopathy . The present animal model was evaluated to be useful in the studies on myelopathy . A new clinical evidence-based gene-environment interaction model of depression . In our current understanding of mood disorders , the role of genes is diverse including the mediation of the effects of provoking and protective factors . Different or partially overlapping gene sets play a major role in the development of personality traits including also affective temperaments , in the mediation of the effects of environmental factors , and in the interaction of these elements in the development of depression . Certain genes are associated with personality traits and temperaments including e.g. , neuroticism , impulsivity , openness , rumination and extroversion . Environmental factors consist of external ( early and provoking life events , seasonal changes , social support etc. ) and internal factors ( hormones , biological rhythm generators , comorbid disorders etc ) . Some of these environmental factors , such as early life events and some prenatal events directly influence the development of personality traits and temperaments . In the NEWMOOD cohort polymorphisms of the genes of the serotonin transporter , P08908 , P28222 and 5- Q13049 and endocannabinoid P21554 receptors , tryptophan hydroxylase , P16220 , P23560 and GIRK provide evidence for the involvement of these genes in the development of depression . Based on their role in this process they could be assigned to different gene sets . The role of certain genes , such as promoter polymorphisms of the serotonin transporter ( 5-HTTLPR ) and P21554 receptor has been shown in more than one of the above factors . Furthermore , gene-gene interactions of these promoters associated with anxiety suggest the application of these polymorphisms in personalized medicine . In this review we introduce a new model including environmental factors , genes , trait and temperament markers based on human genetic studies . 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 . 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 . 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 . 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 . 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 . Autocrine endocannabinoid signaling through P21554 receptors potentiates OX1 orexin receptor signaling . It has been proposed that OX(1) orexin receptors and CB(1) cannabinoid receptors can form heteromeric complexes , which affect the trafficking of OX(1) receptors and potentiate OX(1) receptor signaling to extracellular signal-regulated kinase ( P29323 ) . We have recently shown that OX(1) receptor activity releases high levels of the endocannabinoid 2-arachidonoyl glycerol ( 2-AG ) , suggesting an alternative route for OX(1)-CB(1) receptor interaction in signaling , for instance , in retrograde synaptic transmission . In the current study , we set out to investigate this possibility utilizing recombinant Chinese hamster ovary P04264 cells . 2-AG released from OX(1) receptor-expressing cells acted as a potent paracrine messenger stimulating P29323 activity in neighboring CB(1) receptor-expressing cells . When OX(1) and CB(1) receptors were expressed in the same cells , OX(1) stimulation-induced P29323 phosphorylation and activity were strongly potentiated . The potentiation but not the OX(1) response as such was fully abolished by specific inhibition of CB(1) receptors or the enzyme responsible for 2-AG generation , diacylglycerol lipase ( DAGL ) . Although the results do not exclude the previously proposed OX(1)-CB(1) heteromerization , they nevertheless unequivocally identify DAGL-dependent 2-AG generation as the pivotal determinant of the OX(1)-CB(1) synergism and thus suggest a functional rather than a molecular interaction of OX(1) and CB(1) receptors . DB01257 in paroxysmal nocturnal haemoglobinuria . DB01257 is a monoclonal antibody that binds with high affinity to the complement protein P01031 , preventing terminal complement-mediated intravascular haemolysis in patients with paroxysmal nocturnal haemoglobinuria ( PNH ) . In three well designed clinical trials in patients with PNH , eculizumab blocked serum haemolytic activity and decreased transfusion rates . Pooled data from the three clinical trials demonstrated that eculizumab treatment decreased the overall thromboembolism rate in patients with PNH . DB01257 carries a black box warning for the potential increased risk of meningococcal infections and requires patients to receive the meningococcal vaccine at least 2 weeks before starting treatment . DB01257 is the first drug to be approved by the US FDA for the treatment of PNH and is a novel treatment that offers a new option for patients with PNH . 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 .

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interacts_with DB00712?

multiple_choice

Oral aspirin challenges in patients with a history of intolerance to single non-steroidal anti-inflammatory drugs . Summary Background In the clinical practice patients with a history of acute urticaria induced by a single non-steroidal anti-inflammatory drug ( NSAID ) and seeking for safe alternative drugs generally undergo tolerance tests with alternative NSAIDs that have little or no cyclooxygenase-1 ( P23219 ) enzyme inhibitory activity . This practice does not allow for the detection of single NSAID reactors and may lead to unnecessary avoidance of many potentially useful NSAIDs . OBJECTIVE : Evaluate aspirin challenge as a means to distinguish single from multiple NSAID intolerance in patients with a clinical history of acute urticaria induced by a single NSAID . Methods One hundred and seventeen otherwise normal subjects with a history of acute urticaria following the ingestion of a single NSAID ( pyrazolones ( n=58 ) , nimesulide ( n=17 ) , propionic acid derivatives ( n=13 ) , aryl acetic acid derivatives ( n=14 ) , acetaminophen ( n=9 ) , piroxicam ( n=5 ) , and indometacin ( n=1 ) ) underwent single-blind placebo-controlled oral challenges with aspirin . DB00945 -intolerant subjects underwent further tolerance tests drugs exerting little or no inhibitory activity on P23219 enzyme ( including paracetamol , nimesulide , rofecoxib , tramadol , and floctafenine ) . Results DB00945 induced urticaria in 28/117 ( 24 % ) patients . Five out of 28 ( 18 % ) aspirin reactors did not tolerate alternative NSAID on subsequent oral challenges . Conclusion In subjects with a history of urticaria induced by a single NSAID ( other than aspirin ) the diagnostic workup should start with an aspirin challenge in order to detect single/multiple NSAID reactors . 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 . 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 . Involvement of nuclear factor-kappaB ( NF-kappaB ) signaling in the expression of inducible nitric oxide synthase ( P35228 ) gene in rat P13671 glioma cells . It has been demonstrated from studies using NF-kappaB inhibitors that NF-kappaB may be involved in the P35228 induction stimulated by cytokines and/or lipopolysaccharide ( LPS ) in various cell types and tissues . However , the actions of the inhibitors are less selective and highly cytotoxic . We constructed stable clones of P13671 cells transfected with two types of P25963 mutant genes ( P25963 ( SS --> AA ) ; DB00133 -32/36 to Ala-32/36 , P25963 ( KK --> RR ) ; Lys-21/22 to DB00125 -21/22 ) . P25963 ( SS --> AA ) strongly inhibited ( 1 ) LPS- , IL-1beta- , and P01375 -induced nuclear translocation and DNA binding of NF-kappaB to the kappaB site ; and ( 2 ) P35228 induction stimulated by LPS or IL-1beta plus P01579 . These results indicate that NF-kappaB plays a critical role in cytokines and/or LPS-induced P35228 induction . Surprisingly , similar to the endogenous P25963 , P25963 ( KK --> RR ) was degraded by various stimuli , and proteasome inhibitors blocked this event . These results suggest that another Lys residue(s) , other than Lys-21/22 , may be required for the ligand-induced P25963 degradation by the ubiquitin-proteasome pathway . Anti- P15692 in treatment of central retinal vein occlusion . Macular edema along with macular ischemia is responsible for decreased visual acuity in central retinal vein occlusion . DB00112 ( DB00112 , Genentech ) blocks vascular endothelial growth factor ( P15692 ) induced hyperpermeability of blood vessels . In this prospective case series we investigated the efficacy of anti- P15692 treatment in reduction of central retinal thickness ( CRT ) and improvement in visual acuity ( VA ) . 25 patients were followed up for 12 months and treated monthly with intravitreal bevacizumab . VA and CRT were measured at each visit . Treatment was discontinued as the peak improvement of either parameter was reached and reinstituted in case of deterioration/recurrence of edema . Study endpoints included : VA using ETDRS charts , CRT and number of injections at 12 months . Mean VA from all 25 patients increased by 3.1 logMAR lines ( p < 0.05 compared to baseline ) . The improvement of VA after bevacizumab injection was in correlation with a decrease in CRT In subgroup analyses , patients receiving bevacizumab injection within the first 3 months after CRVO showed an average VA gain of 4.2 logMAR lines . Mean of 4.5 injections was needed to control the disease during the follow-up period . DB00112 treatment was effective in VA and reducing CRT . It appears from subgroup analysis that initiation of treatment early in the course of disease produced better functional outcome . Several injections were needed to control the disease . Regular O75051 examinations and retreatment are advised in order to maintain initially reached VA . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . Immunoregulatory mechanisms underlying prevention of colitis-associated colorectal cancer by probiotic bacteria . BACKGROUND : Inflammatory bowel disease ( Q9UKU7 ) increases the risk of colorectal cancer . Probiotic bacteria produce immunoregulatory metabolites in vitro such as conjugated linoleic acid ( DB01211 ) , a polyunsaturated fatty acid with potent anti-carcinogenic effects . This study aimed to investigate the cellular and molecular mechanisms underlying the efficacy of probiotic bacteria in mouse models of cancer . METHODOLOGY/PRINCIPAL FINDINGS : The immune modulatory mechanisms of VSL#3 probiotic bacteria and DB01211 were investigated in mouse models of inflammation-driven colorectal cancer . Colonic specimens were collected for histopathology , gene expression and flow cytometry analyses . Immune cell subsets in the mesenteric lymph nodes ( MLN ) , spleen and colonic lamina propria lymphocytes ( P06858 ) were phenotypically and functionally characterized . Mice treated with DB01211 or VSL#3 recovered faster from the acute inflammatory phase of disease and had lower disease severity in the chronic , tumor-bearing phase of disease . Adenoma and adenocarcinoma formation was also diminished by both treatments . VSL#3 increased the mRNA expression of P01375 -α , angiostatin and Q07869 γ whereas DB01211 decreased P35354 levels . Moreover , VSL#3-treated mice had increased Q16552 expression in MLN P01730 + T cells and accumulation of Treg P06858 and memory P01730 + T cells . CONCLUSIONS/SIGNIFICANCE : Both DB01211 and VSL#3 suppressed colon carcinogenesis , although VSL#3 showed greater anti-carcinogenic and anti-inflammatory activities than DB01211 . Mechanistically , DB01211 modulated expression of P35354 levels in the colonic mucosa , whereas VSL#3 targeted regulatory mucosal P01730 + T cell responses . Regulation of cyclooxygenase-2 expression by heat : a novel aspect of heat shock factor 1 function in human cells . The heat-shock response , a fundamental defense mechanism against proteotoxic stress , is regulated by a family of heat-shock transcription factors ( HSF ) . In humans Q00613 is considered the central regulator of heat-induced transcriptional responses . The main targets for Q00613 are specific promoter elements ( HSE ) located upstream of heat-shock genes encoding cytoprotective heat-shock proteins ( HSP ) with chaperone function . In addition to its cytoprotective function , Q00613 was recently hypothesized to play a more complex role , regulating the expression of non-HSP genes ; however , the non-canonical role of Q00613 is still poorly understood . Herein we report that heat-stress promotes the expression of cyclooxygenase-2 ( P35354 ) , a key regulator of inflammation controlling prostanoid and thromboxane synthesis , resulting in the production of high levels of prostaglandin-E(2) in human cells . We show that heat-induced P35354 expression is regulated at the transcriptional level via Q00613 -mediated signaling and identify , by in-vitro reporter gene activity assay and deletion-mutant constructs analysis , the P35354 heat-responsive promoter region and a new distal cis-acting HSE located at position -2495 from the transcription start site . As shown by ChIP analysis , Q00613 is recruited to the P35354 promoter rapidly after heat treatment ; by using shRNA-mediated Q00613 suppression and HSE-deletion from the P35354 promoter , we demonstrate that Q00613 plays a central role in the transcriptional control of P35354 by heat . Finally , P35354 transcription is also induced at febrile temperatures in endothelial cells , suggesting that Q00613 -dependent P35354 expression could contribute to increasing blood prostaglandin levels during fever . The results identify P35354 as a human non-classical heat-responsive gene , unveiling a new aspect of Q00613 function . Leukotriene C4 production during hypoxic pulmonary vasoconstriction in isolated rat lungs . Leukotriene inhibitors preferentially inhibit hypoxic pulmonary vasoconstriction in isolated rat lungs . If lipoxygenase products are involved in the hypoxic pressor response they might be produced during acute alveolar hypoxia and a leukotriene inhibitor should block both the vasoconstriction and leukotriene production that occurs in response to hypoxia . We investigated in isolated blood perfused rat lungs whether leukotriene C4 ( LTC4 ) could be recovered from whole lung lavage fluid during ongoing hypoxic vasoconstriction . Lung lavage from individual rats had slow reacting substance ( SRS ) -like myotropic activity by guinea pig ileum bioassay . Pooled lavage ( 10 lungs ) as analyzed by reverse phase high performance liquid chromatography had an ultraviolet absorbing component at the retention time for LTC4 . At radioimmunoassay , and SRS myotropic activity by bioassay . LTC4 was not found during normoxic ventilation , during normoxic ventilation after a hypoxic pressor response , or during vasoconstriction elicited by DB00761 . DB00711 citrate , a leukotriene synthesis blocker , concomitantly inhibited the hypoxic vasoconstriction and LTC4 production . Thus P09917 products may play a role in the sequence of events leading to hypoxic pulmonary vasoconstriction . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . 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 . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . P11362 -induced epithelial to mesenchymal transition through MAPK/PLCγ/ P35354 -mediated mechanisms . Tumour invasion and metastasis is the most common cause of death from cancer . For epithelial cells to invade surrounding tissues and metastasise , an epithelial-mesenchymal transition ( EMT ) is required . We have demonstrated that P11362 expression is increased in bladder cancer and that activation of P11362 induces an EMT in urothelial carcinoma ( UC ) cell lines . Here , we created an in vitro P11362 -inducible model of EMT , and used this model to identify regulators of urothelial EMT . P11362 activation promoted EMT over a period of 72 hours . Initially a rapid increase in actin stress fibres occurred , followed by an increase in cell size , altered morphology and increased migration and invasion . By using site-directed mutagenesis and small molecule inhibitors we demonstrated that combined activation of the mitogen activated protein kinase ( MAPK ) and phospholipase C gamma ( PLCγ ) pathways regulated this EMT . Actin stress fibre formation was regulated by PLCγ activation , and was also important for the increase in cell size , migration and altered morphology . MAPK activation regulated migration and P12830 expression , indicating that combined activation of PLCγ and MAPK is required for a full EMT . We used expression microarrays to assess changes in gene expression downstream of these signalling cascades . P35354 was transcriptionally upregulated by P11362 and caused increased intracellular prostaglandin E(2) levels , which promoted migration . In conclusion , we have demonstrated that P11362 activation in UC cells lines promotes EMT via coordinated activation of multiple signalling pathways and by promoting activation of prostaglandin synthesis . Equitoxic doses of 5-azacytidine and 5-aza-2'deoxycytidine induce diverse immediate and overlapping heritable changes in the transcriptome . BACKGROUND : The hypomethylating agent DB00928 ( 5-Aza-CR ) is the first drug to prolong overall survival in patients with myelodysplastic syndrome ( P43034 ) . Surprisingly , the deoxyribonucleoside analog 5-Aza-2'deoxycytidine ( 5-Aza-CdR ) did not have a similar effect on survival in a large clinical trial . Both drugs are thought to exert their effects after incorporation into DNA by covalent binding of DNA methyltransferase ( P26358 ) . While 5-Aza-CdR is incorporated into only DNA , 5-Aza-CR is also incorporated into RNA . Here , we have analyzed whether this difference in nucleic acid incorporation may influence the capacities of these drugs to regulate the expression of mRNA and microRNAs ( miRNA ) , which may potentially affect the activities of the drugs in patients . METHODOLOGY/PRINCIPAL FINDINGS : A hematopoietic ( HL-60 ; acute myeloid leukemia ) and a solid ( T24 ; transitional cell carcinoma ) cancer cell line were treated with equitoxic doses of 5-Aza-CR and 5-Aza-CdR for 24 hrs , and the immediate ( day 2 ) and lasting ( day 8 ) effects on RNA expression examined . There was considerable overlap between the RNAs heritably upregulated by both drugs on day 8 but more RNAs were stably induced by the deoxy analog . Both drugs strongly induced expression of cancer testis antigens . On day 2 more RNAs were downregulated by 5-Aza-CR , particularly at higher doses . A remarkable downregulation of miRNAs and a significant upregulation of tRNA synthetases and other genes involved in amino acid metabolism was observed in T24 cells . CONCLUSIONS/SIGNIFICANCE : Overall , this suggests that significant differences exist in the immediate action of the two drugs , however the dominant pattern of the lasting , and possible heritable changes , is overlapping . 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 . 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 . 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 . Epigenetics , an early event in the modulation of gene expression by inositol hexaphosphate in ethylnitrosourea exposed mouse lungs . Mechanisms of anticancer effects of inositol hexaphosphate are not fully understood . Epigenetic changes are the early changes in tumorigenesis . DNA methyl transferases , methyl CpG binding proteins , methyl CpG DNA binding domain protein , and histone deacetylases are the major molecules involved in epigenetics . We have shown the effects of IP6 at the molecular level in mouse lungs before the tumor is developed . After 3 mo of ENU exposure , there was no tumor formation , but there was hyperplasia and lymphocytic infiltration in the lungs . Inflammation and DNA damage repair enzymes P35354 and P40692 appear to be upregulated , whereas tumor suppressor gene p16 was downregulated by ENU . On the other hand , ENU exposure more or less upregulated the epigenetic events such as the expressions of P26358 , MeCP2 , Q9UIS9 , and Q13547 . This alteration was reduced by IP6 administration . Results were supported by modulation of global DNA methylation and the modulation of promoter CpG methylation of p16 , P40692 , and P35354 genes . Hence , this study indicates the possible role of epigenetics at the early stage of tumor development and in the regulation of gene expression by IP6 before the onset of ENU-induced lung tumors . DB01373 -dependent proteinase activity in root cultures of Arabidopsis . The presence of calcium-dependent proteinase in plants was investigated using soluble proteins from Arabidopsis root cultures . DB01373 increased the proteolytic activity by two to three fold . DB01373 -stimulated proteolytic activity was inhibited by different protease inhibitors . The highest inhibition ( 79 % ) of calcium-dependent proteolytic activity was observed in the presence of leupeptin . Thirty-eight percent of calcium-dependent proteolytic activity was inhibited by E-64 , an inhibitor of cysteine proteinases . P62158 did not have any effect on calcium-dependent as well as calcium-independent proteolytic activity in the cellular extract . These results provide the first evidence for the presence of calcium activated proteinase in plants and open new avenues to investigate the mode of calcium action in plants .

MH_train_1418

MH_train_1418

MH_train_1418

interacts_with DB00333?

multiple_choice

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 . 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 . 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 . Expression of two variants of the human mu opioid receptor mRNA in SK-N-SH cells and human brain . A partial mu opioid receptor gene was isolated from a human genomic library using a mouse delta opioid receptor cDNA as a probe . Using information from this genomic clone and the published human mu receptor , P35372 , a cDNA was isolated from SK-N-SH mRNA that codes for a variant of the P35372 mRNA , MOR1A . The presence of MOR1A is also shown in human brain using RT-PCR . MOR1A differs from P35372 in that the 3' terminal intron has not been removed . An in-frame termination codon is found four amino acids after the 5' consensus splice site , making MOR1A eight amino acids shorter than P35372 . Both receptors show similar ligand binding and coupling to DB02527 in CHO- P04264 cells . The C-terminal differences between P35372 and MOR1A could have effects on receptor coupling or receptor transport and localization . 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 . The anti-cancer drug camptothecin inhibits elongation but stimulates initiation of RNA polymerase II transcription . DB04690 is a widely used anti-tumor drug that specifically inhibits P11387 . It is believed that topoisomerase I participates in the process of transcription by relaxing torsional stress induced in the duplex DNA by the elongating RNA polymerase . We have assessed the effects of camptothecin on RNA polymerase II transcription from the dihydrofolate reductase ( P00374 ) gene in Chinese hamster ovary ( CHO ) cells . Using in vivo [3H]uridine pulse labeling and in vitro nuclear run-on techniques to estimate relative rates of transcription , it was found that camptothecin stimulated RNA synthesis from promoter-proximal sequences of the P00374 gene , while transcription from promoter-distal sequences was reduced . Furthermore , camptothecin caused a significant accumulation of RNA polymerases in the 5'-end of the P00374 gene . The effect of camptothecin on transcription was reversible , resulting in a wave of RNA synthesis recovery in a 5' to 3' direction through the P00374 gene following a chase with camptothecin-free medium . We conclude that camptothecin stimulates initiation but inhibits elongation of the RNA polymerase II transcribed P00374 gene . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . Genome-wide gene expression analysis reveals a dynamic interplay between luteotropic and luteolytic factors in the regulation of corpus luteum function in the bonnet monkey ( Macaca radiata ) . Although LH is essential for survival and function of the corpus luteum ( CL ) in higher primates , luteolysis occurs during nonfertile cycles without a discernible decrease in circulating LH levels . Using genome-wide expression analysis , several experiments were performed to examine the processes of luteolysis and rescue of luteal function in monkeys . Induced luteolysis with P30968 antagonist ( DB00050 ) resulted in differential regulation of 3949 genes , whereas replacement with exogenous LH ( DB00050 plus LH ) led to regulation of 4434 genes ( 1563 down-regulation and 2871 up-regulation ) . A model system for prostaglandin ( PG ) F(2alpha)-induced luteolysis in the monkey was standardized and demonstrated that P49763 (2alpha) regulated expression of 2290 genes in the CL . Analysis of the LH-regulated luteal transcriptome revealed that 120 genes were regulated in an antagonistic fashion by P49763 (2alpha) . Based on the microarray data , 25 genes were selected for validation by real-time RT-PCR analysis , and expression of these genes was also examined in the CL throughout the luteal phase and from monkeys treated with human chorionic gonadotropin ( hCG ) to mimic early pregnancy . The results indicated changes in expression of genes favorable to P49763 (2alpha) action during the late to very late luteal phase , and expressions of many of these genes were regulated in an opposite manner by exogenous hCG treatment . Collectively , the findings suggest that curtailment of expression of downstream LH-target genes possibly through P49763 (2alpha) action on the CL is among the mechanisms underlying cross talk between the luteotropic and luteolytic signaling pathways that result in the cessation of luteal function , but hCG is likely to abrogate the P49763 (2alpha)-responsive gene expression changes resulting in luteal rescue crucial for the maintenance of early pregnancy . Linkage assignment of eleven genes to the porcine genome . We report comparative linkage mapping of eleven genes in the swine genome by RFLP analysis . These genes include : Acid phosphatase type 5 ( P13686 ) , Cholecystokinin Type B Receptor ( P32239 ) , Antibiotic Peptide ( P49913 ) , P01308 -like Growth Factor 1 Receptor ( P08069 ) , Integrin Alpha M ( P11215 ) , Integrin Beta 2 ( ITGbeta2 ) , Opioid Receptor Mu-1 ( P35372 ) , Pro-hormone Converter ( PC1/3 ) , DB00162 Binding Q12988 ( P10745 ) , Ribosomal DNA ( RNR1 ) , and Zona Pellucida Glycoprotein 1 ( P60852 ) . The P32239 and ITGbeta2 loci define the ends of the linkage groups on Chromosomes ( Chro ) ( SSC ) 9p and 13qter , respectively . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . Insights into antifolate resistance from malarial P00374 -TS structures . Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) is an important target of antimalarial drugs . The efficacy of this class of P00374 -inhibitor drugs is now compromised because of mutations that prevent drug binding yet retain enzyme activity . The crystal structures of PfDHFR-TS from the wild type ( TM4/8.2 ) and the quadruple drug-resistant mutant ( V1/S ) strains , in complex with a potent inhibitor WR99210 , as well as the resistant double mutant ( P04264 P21554 ) with the antimalarial pyrimethamine , reveal features for overcoming resistance . In contrast to pyrimethamine , the flexible side chain of WR99210 can adopt a conformation that fits well in the active site , thereby contributing to binding . The single-chain bifunctional PfDHFR-TS has a helical insert between the P00374 and TS domains that is involved in dimerization and domain organization . Moreover , positively charged grooves on the surface of the dimer suggest a function in channeling of substrate from TS to P00374 active sites . These features provide possible approaches for the design of new drugs to overcome antifolate resistance . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . delta(9)- DB00470 increases nerve growth factor production by prostate PC-3 cells . Involvement of P21554 cannabinoid receptor and P04049 . Cannabinoids , the active components of marihuana , exert a variety of effects in humans . Many of these effects are mediated by binding to two types of cannabinoid receptor , P21554 and CB2 . Although P21554 is located mainly in the central nervous system , it may also be found in peripheral tissues . Here , we study the effect of cannabinoids in the production of nerve growth factor by the prostate tumor cell line PC-3 . We show that addition of Delta(9)-tetrahydrocannabinol to PC-3 cells stimulated nerve growth factor production in a dose-dependent and time-dependent manner . Maximal effect was observed at 0.1 microM Delta(9)-tetrahydrocannabinol and 72 h of treatment . Stimulation was reversed by the P21554 antagonists AM 251 and SR 1411716A . Pre-treatment of cells with pertussis toxin also prevented the effect promoted by Delta(9)-tetrahydrocannabinol . These results indicate that Delta(9)-tetrahydrocannabinol stimulation of nerve growth factor production in these cells was mediated by the cannabinoid P21554 receptor . The implication of P04049 activation in the mode of action of Delta(9)-tetrahydrocannabinol is also suggested . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Selective inhibitors of Q02750 /ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells . The nonsteroidal anti-inflammatory drug ( NSAID ) sulindac prevents experimental colon cancer and can regress precancerous polyps in humans . Sulindac sulfide inhibits cyclooxygenase ( P36551 ) -mediated prostaglandin synthesis and retards the growth of cultured colon cell lines primarily by inducing apoptosis . Given the known role of mitogen-activated protein kinase ( MAPK ) in signal transduction and the regulation of cell survival and death , we determined the effect of sulindac sulfide on MAPK activation , P35354 expression , and apoptosis induction in HCA-7 human colon cancer cells . Sulindac sulfide treatment was associated with activation of ERKp44/42 and p38 MAPK in a dosage- and time-dependent manner , and also activated upstream MEK . Similar results were seen in HCT-15 cells and also with the selective P35354 inhibitor NS398 . ERKp44/42 and p38 activation were accompanied by an induction of P35354 protein expression . Selective inhibitors of sulindac sulfide-induced ERKp44/42 ( PD98059 ) and p38 MAPK ( SB203580 ) activation also suppressed the induction of P35354 by this NSAID . Furthermore , both MAPK inhibitors significantly augmented sulindac sulfide-induced apoptosis , as did suppression of constitutive P35354 using antisense oligonucleotides . In conclusion , MEK/ P29323 and p38 MAPK activation mediate P35354 induction by sulindac sulfide . Selective inhibitors of these MAPKs potentiate apoptosis induction by this NSAID , suggesting a novel strategy for the prevention or treatment of colorectal cancer . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Dimethylsulfoxide and conjugated linoleic acids affect bovine embryo development in vitro . Conjugated linoleic acids ( DB01211 ) are employed to overcome the bovine periparturitional negative energy balance . Especially of interest are trans10,cis12 -linoleic acid ( t10c12- DB01211 ) and cis9,trans11-linoleic acid ( c9t11- DB01211 ) . Their impact on embryonic development , though , is not clear . Here , effects of both above-mentioned DB01211 on bovine in vitro-produced embryos were assessed . Zygotes ( n=2098 ) were allocated to one of seven groups : cultured with 50 or 100µM of either c9t11- DB01211 or t10c12- DB01211 , with 14 or 28mM DB01093 or without supplement ( control ) . Messenger RNA analysis of target gene transcripts ( P08069 , P18065 , P22692 , P23786 , P09110 , P42765 , P49327 , O00767 ) via RT-qPCR was performed in single blastocysts . Cleavage rates did not differ , whereas development rates were decreased in both t10c12-supplemented groups in comparison to the unsupplemented group ( 31.7 % ±2.2 control vs 20.2 % ±2.0 50µM t10c12 vs 21.0 % ±2.8 100µM t10c12 ) . Compared with the unsupplemented group , O00767 was expressed at a lower level in embryos cultured with 50µM c9t11- DB01211 . The relative amount of several transcripts was increased in embryos cultured with 14mM DB01093 in comparison to those that developed in the presence of 50µM t10c12- DB01211 ( P18065 , P09110 , P23786 , P49327 , O00767 ) or 50µM c9t11- DB01211 ( P08069 , P18065 , P09110 , P23786 , P49327 , O00767 ) . The molecular analyses show that DB01211 influence embryonic fat metabolism . 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 . Cytokinins control endocycle onset by promoting the expression of an P25054 /C activator in Arabidopsis roots . Plant roots respond to various internal and external signals and adjust themselves to changes of environmental conditions . In the root meristem , stem cells produce daughter cells that continue to divide several times . When these latter cells reach the transition zone , they stop dividing and enter the endocycle , a modified cell cycle in which DNA replication is repeated without mitosis or cytokinesis . The resultant DNA polyploidization , named endoreduplication , is usually associated with an increase of nuclear and cell volume and with cell differentiation . At the transition zone , cytokinin signaling activates two transcription factors , type-B ARABIDOPSIS RESPONSE REGULATOR 1 ( P49407 ) and ARR12 , and induces SHY2/IAA3 , a member of the Aux/IAA family of auxin signaling repressors . This inhibits auxin signaling and reduces the expression of auxin efflux carriers , resulting in cell division arrest . Such counteracting actions of two hormones are assumed to determine meristem size . However , it remains unknown whether cytokinins additionally control meristem size through an auxin-independent pathway . Here we show that , in Arabidopsis , the cytokinin-activated P32121 directly upregulates the expression of CCS52A1 , which encodes an activator of an E3 ubiquitin ligase , anaphase-promoting complex/cyclosome ( P25054 /C ) , thereby promoting the onset of the endocycle and restricting meristem size . Our genetic data revealed that CCS52A1 function is independent of SHY2-mediated control of auxin signaling , indicating that downregulation of auxin signaling and P25054 /C-mediated degradation of cell-cycle regulators cooperatively promote endocycle onset , and thus fine tune root growth . DB02745 adenosine tetraphosphate ( Up4A ) is a strong inductor of smooth muscle cell migration via activation of the P41231 receptor and cross-communication to the PDGF receptor . The recently discovered dinucleotide uridine adenosine tetraphosphate ( Up(4)A ) was found in human plasma and characterized as endothelium-derived vasoconstrictive factor ( EDCF ) . A further study revealed a positive correlation between Up(4)A and vascular smooth muscle cell ( VSMC ) proliferation . Due to the dominant role of migration in the formation of atherosclerotic lesions our aim was to investigate the migration stimulating potential of Up(4)A . Indeed , we found a strong chemoattractant effect of Up(4)A on VSMC by using a modified Boyden chamber . This migration dramatically depends on osteopontin secretion ( P10451 ) revealed by the reduction of the migration signal down to 23 % during simultaneous incubation with an P10451 -blocking antibody . Due to inhibitory patterns using specific and unspecific purinoreceptor inhibitors , Up(4)A mediates it 's migratory signal mainly via the P2Y(2) . The signaling behind the receptor was investigated with luminex technique and revealed an activation of the extracellular signal-regulated kinases 1 and 2 ( P27361 /2 ) pathway . By use of the specific PDGF receptor ( P09619 ) inhibitor AG1296 and siRNA technique against P09619 -β we found a strongly reduced migration signal after Up(4)A stimulation in the P09619 -β knockdown cells compared to control cells . In this study , we present substantiate data that Up(4)A exhibits migration stimulating potential probably involving the signaling cascade of Q02750 and P27361 /2 as well as the matrix protein P10451 . We further suggest that the initiation of the migration process occurs predominant through direct activation of the P2Y(2) by Up(4)A and via transactivation of the P09619 . Polyamines and membrane transporters . In recent years , our understanding of the importance of membrane transporters ( MTs ) in the disposition of and response to drugs has increased significantly . MTs are proteins that regulate the transport of endogenous molecules and xenobiotics across the cell membrane . In mammals , two super-families have been identified : DB00171 -binding cassette ( DB01048 ) and solute carrier ( O00585 ) transporters . There is evidence that MTs might mediate polyamines ( PA ) transport . PA are ubiquitous polycations which are found in all living cells . In mammalian cells , three major PA are synthesised : putrescine , spermidine and spermine ; whilst the decarboxylated arginine ( agmatine ) is not produced by mammals but is synthesised by plants and bacteria . In addition , research in the PA field suggests that PA are transported into cells via a specific transporter , the polyamine transport system(s) ( Q03393 ) . Although the Q03393 has not been fully defined , there is evidence that some of the known MTs might be involved in PA transport . In this mini review , eight O00585 transporters will be reviewed and their potential to mediate PA transport in human cells discussed . These transporters are O15245 , O15244 , O75751 , Q96FL8 , P30825 , P08195 , SLC12A8A , and Q86VW1 . Preliminary data from our laboratory have revealed that O15245 might be involved in the PA uptake ; in addition to one member of ABC superfamily ( P08183 protein ) might also mediate the efflux of polyamine like molecules . Polymorphism in folate- and methionine-metabolizing enzyme and aberrant CpG island hypermethylation in uterine cervical cancer . OBJECTIVE : This study was conducted to explore the association between the CpG island hypermethylation of tumor-associated genes and the polymorphisms of methyl group metabolizing enzymes in uterine cervical cancer . METHODS : We analyzed CpG island hypermethylation in 15 genes ( P25054 , CDH1 , P35354 , P53355 , P49789 , P09211 , HLTF1 , hMLH1 , P16455 , p14 , p16 , RASSF1A , Q13761 , P07996 , and P35625 ) and its association with the methylene- DB00116 reductase ( P42898 ) C677T and A1298C and the methionine synthase ( MS ) A2756G polymorphisms in 82 Korean women with uterine cervical cancer . RESULTS : All uterine cervical cancer samples had at least one gene methylated . The average number of methylated genes was lower in patients with the heterozygous genotype of P42898 and MS than in those with the common homozygous genotype , although this difference was not significant . The P42898 677 CT genotype was significantly associated with the decreased promoter hypermethylation of O(6)-methylguanine DNA methyltransferase ( P16455 ) ( OR = 0.22 , 95 % confidence interval ( CI ) 0.07-0.70 , P = 0.011 ) . However , the P42898 C677T and A1298C and the MS A2756G polymorphisms were not associated with an increased risk of uterine cervical cancer . CONCLUSION : These findings suggest that there is a possible interaction between epigenetic and genetic factors in uterine cervical cancer .

MH_train_1419

MH_train_1419

MH_train_1419

interacts_with DB08899?

multiple_choice

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 . 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 . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . DB08439 inhibits apoptosis in acute myocardial infarction due to permanent coronary ligation but not due to ischemia-reperfusion . PURPOSE : Myocardial ischemia induces cyclooxygenase 2 ( P35354 ) expression . We evaluated the effects of parecoxib , a P35354 inhibitor , in 2 different mouse models of myocardial ischemia : permanent left coronary artery ligation ( PI ) and transient ligation ( 30 minutes ischemia ) followed by reperfusion ( I/R ) . METHODS : Forty adult male Institute of Cancer Research mice underwent PI ( n = 24 ) or I/R ( n = 16 ) , followed by randomization to parecoxib ( 0.75 mg/kg intraperitoneal daily ) or normal saline for 7 days . RESULTS : DB08439 significantly reduced apoptosis [ 0.8 % vs. 3.4 % ( saline ) , P < 0.001 ] and 7-day mortality [ 0 % vs. 57 % ( saline ) , P = 0.040 ] in the PI group but showed no benefit in the I/R group . DB08439 -treated mice also exhibited greater fractional shortening in the PI group [ 22 % vs. 14 % ( saline ) , P = 0.045 ) but not in the I/R group . DB08439 did not affect infarct size in either group . CONCLUSIONS : P35354 may play a pivotal role in mediating apoptosis in the ischemic peri-infarct myocardium that is not reperfused after infarct . 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 . Induction of cytokine gene expression in human thyroid epithelial cells irradiated with HZE particles ( iron ions ) . Gene expression profiles were examined using cDNA microarray technology in human thyroid epithelial ( Htori-3 ) cells exposed to a low , non-toxic dose ( 10 cGy ) of radiation from HZE particles in the form of iron ions in the absence or presence of selenomethionine ( SeM ) . A total of 215 genes were differentially regulated 2 h after exposure to a 10-cGy dose of iron-ion radiation . In the microarray analysis , SeM had profound effects on the radiation-induced expression of several specific genes , which includes P00749 , P17936 , P15328 , P15291 and P02452 . Of particular interest to us was a gene cluster , " secreted proteins " , that was up-regulated after radiation exposure . Seven up-regulated genes of this gene cluster fall within the chemokine/cytokine gene cluster , namely , P09341 , P19875 , P05231 , P20809 , P10145 , Q13007 and TGFbeta2 . In microarray studies , the radiation-induced up-regulated expression of some these genes encoding cytokine/chemokine proteins was significantly decreased by SeM treatment . For P10145 , TGFbeta2 , P09341 and P19875 , these observations were validated by qPCR techniques . It is concluded that SeM can regulate ionizing radiation-induced gene expression and may serve as an effective countermeasure for some of the acute inflammatory/immune responses induced by low-dose HZE-particle radiation . 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 . The anti-interleukin-6 antibody siltuximab down-regulates genes implicated in tumorigenesis in prostate cancer patients from a phase I study . BACKGROUND : P05231 ( P05231 ) is associated with prostate cancer morbidity . In several experimental models , P05231 has been reported to have anti-apoptotic and pro-angiogenic effects . DB09036 ( CNTO 328 ) is a monoclonal anti- P05231 antibody which has been successfully applied in several models representing prostate cancer . This study was designed to assess preliminary safety of siltuximab in patients with early prostate cancer . PATIENTS AND METHODS : Twenty patients scheduled to undergo radical prostatectomy received either no drug or siltuximab ( 6 mg/kg , five patients per group with administration once , two times , and three times prior to surgery ) . Blood samples were collected for pharmacokinetic and pharmacodynamic analyses . Expression of elements of P05231 signaling pathways was analyzed in tumor tissue by immunohistochemistry . Gene analysis in tumor specimens was performed with the DASL array . RESULTS : No adverse events related to siltuximab were observed . Patients treated with siltuximab presented with higher levels of proliferation and apoptosis markers . Following a single dose , serum concentrations of siltuximab declined in a biexponential manner . This study revealed a decrease in phosphorylation of Stat3 and Q8TCB0 / Q8NFH3 mitogen-activated protein kinases . In addition , gene expression analyses indicate down-regulation of genes immediately downstream of the P05231 signaling pathway and key enzymes of the androgen signaling pathway . CONCLUSIONS : Preliminary safety of siltuximab is favorable . Future studies in which siltuximab could be combined with androgen-deprivation therapy and experimental therapies in advanced prostate cancer are justified . 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 . Vitamin D mediates its action in human colon carcinoma cells in a calcium-sensing receptor-dependent manner : downregulates malignant cell behavior and the expression of thymidylate synthase and survivin and promotes cellular sensitivity to DB00544 . Vitamin D ( VD ) protects against colon carcinogenesis by mechanisms not fully understood . We had earlier reported on the similarity in the biologic action of VD and that of the calcium-sensing receptor ( P41180 ) in human colon carcinoma cells . At the molecular level , the P41180 gene contains 2 VD response elements and VD stimulates the expression of P41180 . In this study , we investigated on the relationship between VD action and P41180 function . We determined and compared the action of VD in human colon carcinoma cells ( P35520 , Moser , Caco-2 and HCT116 ) and their P41180 knocked-down counterparts . VD inhibited cellular proliferation , cellular invasion , and anchorage-independent growth and stimulated the expression of P38936 /Waf1 but not in P41180 knocked-down cells . These results demonstrate , for the first time , that the known tumor-suppressive function of VD requires functional P41180 and knocking down P41180 expression abrogated this function of VD . We recently reported that activation of P41180 in human colon carcinoma cells downregulated the expression of thymidylate synthase ( TS ) and survivin and promoted a significant increase in sensitivity to cytotoxic drugs . We now demonstrate , for the first time , that VD suppressed the expression of TS and survivin , TS and survivin gene transcriptional activities and promoted a cytotoxic response to DB00544 in a P41180 -dependent manner . Ectopic expression of wild-type P41180 in colon carcinoma cells also inhibited the expression of TS and survivin and enhanced cellular sensitivity to DB00544 . VD , however , could no longer enhance cellular sensitivity to DB00544 in cells overexpressing P41180 . Regional expression of prostaglandin E2 and F2alpha receptors in human myometrium , amnion , and choriodecidua with advancing gestation and labor . The change from uterine quiescence to enhanced contractile activity may be due to the differential expression of prostaglandin receptors within the myometrium and fetal membranes , in a temporal and topographically distinct manner . To address this question , we determined the localization and expression of the DB00917 receptor subtypes ( P34995 -4 ) and the PGF2alpha receptor ( P43088 ) in paired upper and lower segment myometrium , amnion , and choriodecidual samples throughout human pregnancy , with and without labor . All receptor subtypes were found throughout the muscle layers in both the upper and lower uterine segments , colocalizing with alpha smooth muscle actin . A change in intracellular localization was observed at term labor , where P34995 and P35408 were predominately associated with the nucleus . Minimal changes in the expression of the DB00917 and PGF2alpha receptor subtypes were observed with gestational age , labor , or between the upper and lower myometrial segments . Receptor expression in maternal and fetal tissues differed between the receptor subtypes ; P34995 and P35408 were predominately expressed in the fetal membranes , PTGER2 was greatest in the myometrium , whereas P43115 and P43088 were similarly expressed in the myometrium and fetal membranes . Myometrial activation through the prostaglandin receptors is perhaps more subtle and may be mediated by a balance between one or several of the prostaglandin receptor subtypes together with other known contraction associated proteins . Lack of coordination in receptor expression between the myometrium and fetal membranes may indicate different regulatory mechanisms between these tissues , or it may suggest a function for these receptors in the amnion and choriodecidua that is independent of that seen in the myometrium . The anti-inflamm-aging and hepatoprotective effects of huperzine A in D-galactose-treated rats . Oxidative stress contributes to a chronic inflammatory process referred to as " inflamm-aging " . P22303 inhibitors ( AChEI ) can enhance cholinergic transmission and act as anti-inflammatory agents via immunocompetent cells expressing α-7 acetylcholine receptors ( AChR ) . The present study explores the possible role of huperzine A , a reversible and selective AChEI , against D-gal-induced oxidative damage , cell toxicity and inflamm-aging in rat livers . In two-month-old rats with normal liver function , an 8-week administration of D-gal ( 300 mg/kg subcutaneously ( s.c. ) injected ) , significantly increased hepatic impairment , ROS generation and oxidative damage , hepatic senescence , nuclear factor-kappa B ( NF-κB ) activation and inflammatory responses . An 8-week co-administration of both D-gal ( 300 mg/kg s.c. ) and huperzine A ( 0.1 mg/kg s.c. ) not only significantly decreased hepatic function impairment , ROS generation , oxidative damage , but also suppressed inflamm-aging by inhibiting hepatic replicative senescence , P22303 activity , IκBα degradation , NF-κB p65 nuclear translocation and inflammatory responses . The expression levels of pro-inflammatory cytokine mRNA and proteins , such as TNFα , IL-1β and P05231 decrease significantly , and the protein levels of the anti-inflammatory cytokine P22301 display an obvious increase . These findings indicated that D-gal-induced hepatic injury and inflamm-aging in the rat liver was associated with the development of a pro-inflammatory phenotype in this organ . D-gal induced damage-associated molecular patterns ( DAMPs ) because oxidative damages might play an important role in D-gal-induced hepatic sterile inflammation . DB04864 exhibited protective effects against D-gal-induced hepatotoxicity and inflamm-aging by inhibiting P22303 activity and via the activation of the cholinergic anti-inflammatory pathway . The huperzine A mechanism might be involved in the inhibition of DAMPs-mediated NF-κB nuclear localization and activation . P10275 and nutrient signaling pathways coordinate the demand for increased amino acid transport during prostate cancer progression . L-Type amino acid transporters such as Q01650 and O75387 mediate the uptake of essential amino acids . Here , we report that prostate cancer cells coordinate the expression of Q01650 and O75387 to maintain sufficient levels of leucine needed for mTORC1 signaling and cell growth . Inhibiting O43561 function was sufficient to decrease cell growth and mTORC1 signaling in prostate cancer cells . These cells maintained levels of amino acid influx through androgen receptor-mediated regulation of O75387 expression and P18848 regulation of Q01650 expression after amino acid deprivation . These responses remained intact in primary prostate cancer , as indicated by high levels of O75387 in primary disease , and by increased levels of Q01650 after hormone ablation and in metastatic lesions . Taken together , our results show how prostate cancer cells respond to demands for increased essential amino acids by coordinately activating amino acid transporter pathways vital for tumor outgrowth . 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 . The effect of progesterone on genes involved in preterm labor . The decidua is known to be a major source of intrauterine PGF2α during late gestation and labor , and inflammatory cytokines , including IL-1β , P05231 , and P10145 , are elevated in spontaneous preterm deliveries . In the present study , to elucidate how progesterone blocks the pathways associated with preterm birth , we determined the effects of P4 on the expression of PTGS-2 and P43088 mRNA in human decidua fibroblast cells , as well as the genes , using microarray analysis . Senescence was induced in primary cultured human decidual cells treated with IL-1β . The IL-1β treatment implicated by microarray analysis increased gene expression levels of PTGS-2 , P43088 , NFκ-B p65 , Q16552 , and P10145 . In contrast , P4+IL-1β decreased the expression levels of all of these genes in comparison to treatment with IL-1β alone ( p < 0.05 ) . IL-1β also increased the proportion of SA-β-gal-positive cells . Treatment with IL-1β also increased the P38936 protein level in comparison to cells treated either with the vehicle or P4 . Neither the P38936 protein level nor the number of SA-β-gal-positive cells was increased in normal endometrial glandular cells by IL-1β ( p < 0.05 ) . Our studies demonstrated that P4 changes the level of gene expression in a manner that favors an anti-inflammatory milieu . Because P10145 appears to be the cytokine whose expression is most significantly modulated by P4 , further studies evaluating P10145 as a therapeutic target are needed . An intact NF-kappaB pathway is required for histone deacetylase inhibitor-induced P55008 arrest and maturation in U937 human myeloid leukemia cells . The role of NFkappaB in regulating P55008 arrest and maturation induced by the histone deacetylase inhibitor sodium butyrate ( NaB ) was examined in human myelomonocytic leukemia cells ( U937 ) . Cells stably transfected with an P25963 " super-repressor " lacking phosphorylation sites necessary for proteasomal degradation exhibited diminished IkBa phosphorylation and NF-kappaB DNA binding upon exposure to TNFalpha When exposed to NaB ( 1 mM ; 48 hr ) or PMA ( 5 nM ; 24 hr ) , IkappaBalphaM cells displayed a marked reduction in P55008 arrest compared to Neo controls . In each case , this was accompanied by a significant reduction in the percentage of cells expressing the differentiation markers CD11a , CD11b , and P05107 . The impairment in NaB-induced maturation in mutant cells was associated with a reciprocal increase in apoptosis . In contrast to impairment in NaB- or PMA-induced NF-kappaB DNA binding , stable expression of the IkappaBalphaM did not modify DNA binding of SP1 or AP2 transcription factors . IkappaBalphaM cells also displayed impairment in NaB- and PMA-mediated induction of p21CIP1 and phosphorylation ( inactivation ) of p34cdc2 , as well as diminished levels of P06400 -bound Q01094 . Finally , the NF-kappaB inhibitor O95347 antagonized NaB- and PMA-related NF-kappaB DNA binding as well as induction of p21CIP1 . Together , these findings suggest that NF-kappaB plays an important functional role in mediating NaB-induced p21CIP1 induction , P55008 arrest , and maturation in human myelomonocytic leukemia cells , and that disruption of the NF-kappaB pathway causes cells to engage an alternative , apoptotic program . 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 . 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 . 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 . DB04725 attenuates quinolinic acid induced Huntington like symptoms : possible behavioral , biochemical and cellular alterations . Cyclo-oxygenase and lipoxygenase enzymes are involved in arachidonic acid metabolism . Emerging evidence indicates that cyclo-oxygenase and lipoxygenase inhibitors prevent neurodegenerative processes and related complications . Therefore , the present study has been designed to explore the neuroprotective potential of licofelone ( dual P35354 /5- P28300 inhibitor ) against quinolinic acid induced Huntington like symptom in rats . Intrastriatal administration of quinolinic acid significantly caused reduction in body weight and motor function ( locomotor activity , rotarod performance and beam walk test ) , oxidative defense ( as evidenced by increased lipid peroxidation , nitrite concentration and decreased endogenous antioxidant enzymes ) , alteration in mitochondrial enzyme complex ( I , II and IV ) activities , raised P01375 -α level and striatal lesion volume as compared to sham treated animals . DB04725 ( 2.5 , 5 and 10 mg/kg ) treatment significantly improved body weight , locomotor activity , rotarod performance , balance beam walk performance , oxidative defense , mitochondrial enzyme complex activities and attenuated P01375 -α level and striatal lesion as compared to control ( quinolinic acid ) . The present study highlights that licofelone attenuates behavioral , biochemical and cellular alterations against quinolinic acid induced neurotoxicity and this could be an important therapeutic avenue to ameliorate the Huntington like symptoms .

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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 . DB00158 uptake by the human syncytiotrophoblast is affected by gestational diabetes , hyperleptinemia , and P01375 -α . BACKGROUND : The mechanisms whereby gestational diabetes mellitus ( GDM ) increases the risk of fetal overgrowth and development of metabolic diseases later in life are likely to involve changes in nutrient supply to the fetus . Hence , in this work , we hypothesize that GDM may affect folic acid ( FA ) supply to the placenta and fetus . METHODS : We compared (3)H-FA uptake by human cytotrophoblasts isolated from normal pregnancies ( normal trophoblasts ; NTB cells ) and GDM pregnancies ( diabetic trophoblasts ; DTB cells ) and investigated the effect of GDM hallmarks on (3)H-FA uptake by BeWo cells . RESULTS : (3)H-FA uptake by NTB and DTB cells was time dependent and acidic pH stimulated . When compared with NTB , (3)H-FA uptake by DTB cells was more sensitive to acidic pH changes and to 5-methyltetrahydrofolate and pemetrexed ( PTX ) inhibition , indicating a proportionally greater involvement of the proton-coupled folate transporter ( Q96NT5 ) . A 4-h exposure of BeWo cells to lipopolysaccharide ( LPS , 1-10 μg/ml ) or to high levels of tumor necrosis factor-α ( P01375 -α , 300 ng/l ) significantly reduced (3)H-FA uptake . Moreover , hyperleptinemic conditions ( 100 ng/ml leptin ) decreased (3)H-FA uptake by BeWo cells in a time-dependent manner when compared with normoleptinemic conditions ( 1 ng/ml leptin ) . CONCLUSION : GDM modulates (3)H-FA uptake by the syncytiotrophoblast , and leptin as well as P01375 -α downregulate it . 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 . 5-Hydroxytryptamine induces cyclooxygenase-2 in rat vascular smooth muscle cells : Mechanisms involving Src , PKC and MAPK activation [ corrected ] . Considering the importance of 5-hydroxytryptamine ( 5-HT ) and cyclooxygenase ( P36551 ) products in vascular pathology , we investigated the effects of 5-HT on P36551 expression in rat vascular smooth muscle cells ( VSMCs ) , and to provide mechanistic insights into these effects . VSMCs were enzymatically isolated from aortic media of Wistar rats . Incubation of VSMCs with 5-HT for 24h stimulated prostaglandin I(2) production , but this stimulation was completely suppressed by NS-398 , a selective P35354 inhibitor . 5-HT induced transient P35354 , but not P23219 , protein and mRNA expression in concentration- and time-dependent manners . This effect of 5-HT was completely inhibited by sarpogrelate , a 5-HT(2A) receptor antagonist . 5-HT-induced P35354 expression was markedly blunted by Ca(2+) depletion ; GF 109203X , a protein kinase C ( PKC ) inhibitor ; Q99463 , an inhibitor of Src-family tyrosine kinase ( Src ) ; PD 98059 , an inhibitor of extracellular signal-regulated kinase ( P29323 ) activation ; SB 203580 , an inhibitor of p38 mitogen-activated protein kinase ( MAPK ) ; and SP 600125 , an inhibitor of c-Jun N-terminal kinase ( JNK ) . 5-HT activated P29323 and p38 MAPK , followed by JNK activation . Q99463 inhibited these activations , while GF 109203X inhibited only JNK activation . Furthermore , PD 98059 inhibited JNK activation . These results suggest that 5-HT induces P35354 expression in rat VSMCs , and that PKC , Src , and MAPK activation are each essential for the full expression of P35354 pathways . Effects of selective P35354 and 5- P28300 inhibition on prostaglandin and leukotriene synthesis in ductal pancreatic cancer in Syrian hamster . Selective inhibition of eicosanoid synthesis seems to decrease carcinogenesis , however , the effect on liver metastasis in pancreatic cancer is still unknown . Ductal pancreatic adenocarcinoma was chemically induced by weekly injection of N-nitrosobis-2-oxopropylamine ( BOP ) in Syrian hamster . Animals received selective inhibition of cyclooxygenase-2 ( DB00482 ) and P09917 ( DB00744 ) . In week 33 , hamsters were sacrificed and incidence of pancreatic carcinomas as well as liver metastases were examined . Furthermore , size and number of liver metastases per animal were determined and concentration of PGF1alpha , DB00917 and leukotrienes was measured in hepatic and pancreatic tissue . Combined therapy ( DB00482 + DB00744 ) significantly decreased incidence , number and size of liver metastases . Furthermore extra- and intrametastatic concentration of DB00917 was reduced by this treatment in hepatic tissue . Single Cox-2-inhibition ( DB00482 ) decreased intrametastatic hepatic PGF1alpha and DB00917 concentration while PGF1alpha concentration was reduced in non-metastatic liver ( nml ) . Moreover 5- P28300 -inhibition ( DB00744 ) decreased intrametastatic DB00917 concentration as well as PGF1alpha and DB00917 in nml . In pancreatic carcinomas highest LT-concentration was found after combined treatment and this therapy group was the only one revealing a significantly higher amount of LTs in carcinomas compared to tumour-free tissue . Hepatic LT-concentration was significantly lower in the control groups than in nml of the tumour groups . Combination of Cox-2-inhibition and 5-Lox-inhibition might be a suitable adjuvant therapy to prevent liver metastasis in human ductal pancreatic adenocarcinoma . 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 . [ Association and interaction of AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 and P35354 genes on the risk of hypertension in Antioquian population ] . INTRODUCTION : Hypertension is a multifactorial disease influenced by genetic and environmental components , with its prevalence varying across ethnic groups . Manifold studies on blood pressure regulatory system genes have been carried out -such as the renin-angiotensin-aldosterone system , the sympathetic nervous system , endothelial factor , and sodium balance- , but the results yielded were inconsistent among populations . OBJECTIVES : To evaluate the effect of both variants in genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 P35354 , and the result of the individual ancestry component on hypertension and blood pressure levels among population in Antioquia . METHODS AND MATERIALS : 107 cases and 253 controls were genotyped for 12 variants on genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 y P35354 , and for 20 ancestry informative markers . The association of polymorphisms and their interactions , and the association of ancestral genetic composition with hypertension and blood pressure levels were examined . RESULTS : Genes P35612 , rs4852706 ( OR=3.0 ; p=0.023 ) ; P21728 , rs686 ( OR=0.38 ; p=0.012 ) and P07550 , rs1042718 ( OR=10.0 ; p=0.008 ) ; as well as genotypic combinations of P21728 and P30556 ; AGT and P35611 ; and P35611 to P20020 and P35354 were associated to hypertension . The Amerindian ancestry component was associated to some decrease in diastolic blood pressure . CONCLUSION : Variants on genes P35612 , P21728 , P07550 , P30556 , AGT , P35611 , P20020 and P35354 individually or interacting , are associated to hypertension . The Amerindian ancestry component has an effect on blood pressure . 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 . DB00158 and thiamine transporters mediated by facilitative carriers ( P41440 -3 and Q96NT5 ) and folate receptors . The reduced folate carrier ( P41440 , P41440 ) , thiamine transporter-1 ( O60779 , O60779 ) and thiamine transporter-2 ( Q9BZV2 , Q9BZV2 ) evolved from the same family of solute carriers . P41440 transports folates but not thiamine . O60779 and Q9BZV2 transport thiamine but not folates . P41440 and O60779 deliver their substrates to systemic tissues ; Q9BZV2 mediates intestinal thiamine absorption . The proton-coupled folate transporter ( Q96NT5 , Q96NT5 ) is the mechanism by which folates are absorbed across the apical-brush-border membrane of the proximal small intestine . Two folate receptors ( P15328 and P14207 ) mediate folate transport across epithelia by an endocytic process . DB00158 transporters are routes of delivery of drugs for the treatment of cancer and inflammatory diseases . There are autosomal recessive disorders associated with mutations in genes encoded for Q96NT5 ( hereditary folate malabsorption ) , P15328 ( cerebral folate deficiency ) , O60779 ( thiamine-responsive megaloblastic anemia ) , and Q9BZV2 ( biotin-responsive basal ganglia disease ) . Effects of sulfasalazine and a sulfasalazine analogue on the formation of lipoxygenase and cyclooxygenase products . A sulfasalazine analogue , 5'-(2,4-dichlorobenzoyl)2'-hydroxyphenylacetic acid ( CL 42A ) , potently inhibited the formation of P09917 products ( leukotrienes B4 and C4 and 5-hydroxyeicosatetraenoic acid ) by human leukocytes . Half-maximal inhibition of leukotriene production was obtained with 5 and 10 microM CL 42A after stimulation with serum-treated zymosan or ionophore A23187 , respectively . CL 42A was equipotent to nordihydroguaiaretic acid and about 50 times more potent than sulfasalazine and benoxaprofen in studies on the inhibition of LTB4 formation in leukocyte suspensions stimulated with serum-treated zymosan . Furthermore , CL 42A had no inhibitory effect on the production of 15-hydroxyeicosatetraenoic acid after incubation of human leukocytes with ionophore A23187 in the presence of exogenous arachidonic acid . Sulfasalazine inhibited the synthesis of P09917 products ( 5-hydroxyeicosatetraenoic acid and leukotriene B4 : IC50 250 microM , leukotriene C4 : IC50 100 microM ) in a concentration-dependent manner but had no effect on 15-hydroxyeicosatetraenoic acid formation . The metabolites of sulfasalazine , sulfapyridine and DB00244 , and the isomer , DB00233 , were all less potent than sulfasalazine as inhibitors of leukotriene formation . Both CL 42A ( IC50 20 microM ) and sulfasalazine ( IC50 500 microM ) inhibited the synthesis of thromboxane B2 and hydroxyheptadecatrienoic acid in human platelet suspensions after arachidonic acid stimulation . However , while CL 42A inhibited cyclooxygenase , the inhibitory effect of sulfasalazine was exerted mainly on thromboxane synthase . The platelet formation of 12-hydroxyeicosatetraenoic acid was not inhibited by CL 42A whereas sulfasalazine had a weak inhibitory effect . 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 . Dual effects of histone deacetylase inhibition by trichostatin A on endothelial nitric oxide synthase expression in endothelial cells . Inhibition of histone deacetylases by trichostatin A ( P32119 ) has pleiotropic effects on gene expression . We demonstrated that at low dose ( 0.1 microg ) P32119 increased the P29474 mRNA levels , which was followed by a time- and dose-dependent down-regulation . Cycloheximide , a protein synthesis inhibitor , completely abolished P32119 -induced decrease in P29474 expression , indicating that new protein synthesis is required for the inhibiting effect . DB06693 -- an inhibitor P04035 and geranylgeranylation reaction dose-dependently antagonized P32119 -induced reduction . This mevastatin-mediated antagonism was completely abolished by geranylgeranylpyrophosphate , suggesting that geranylgeranyl modification is needed to activate the P29474 mRNA destabilizing factor -- a mechanism responsible for statin-mediated P29474 upregulation . 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 . DB04630 stimulates vascular smooth muscle cell proliferation via big mitogen-activated protein kinase 1 activation . The nongenomic effects of aldosterone have been implicated in the pathogenesis of various cardiovascular diseases . DB04630 -induced nongenomic effects are attributable in part to the activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) , a classical mitogen-activated protein ( Q96HU1 ) kinase . Q13164 ( Q13164 ) , a newly identified Q96HU1 kinase , has been shown to be involved in cell proliferation , differentiation , and survival . We examined whether aldosterone stimulates Q13164 -mediated proliferation of cultured rat aortic smooth muscle cells ( RASMCs ) . P08235 ( MR ) expression and localization were evaluated by Western blotting analysis and fluorolabeling methods . P27361 /2 and Q13164 activities were measured by Western blotting analysis with the respective phosphospecific antibodies . Cell proliferation was determined by Alamar Blue colorimetric assay . DB04630 ( 0.1 to 100 nmol/L ) dose-dependently activated Q13164 in RASMCs , with a peak at 30 minutes . To clarify whether aldosterone-induced Q13164 activation is an MR-mediated phenomenon , we examined the effect of eplerenone , a selective MR antagonist , on aldosterone-induced Q13164 activation . DB00700 ( 0.1 to 10 micromol/L ) dose-dependently inhibited aldosterone-induced Q13164 activation in RASMCs . DB04630 also stimulated RASMC proliferation , which was inhibited by eplerenone . DB04630 -mediated phenomena were concluded to be attributable to a nongenomic effect because cycloheximide failed to inhibit aldosterone-induced Q13164 activation . Transfection of dominant-negative Q96HU1 kinase/ P29323 kinase 5 ( Q13163 ) , which is an upstream regulator of Q13164 , partially inhibited aldosterone-induced RASMC proliferation , which was almost completely inhibited by MEK inhibitor PD98059 . In addition to the classical steroid activity , rapid nongenomic effects induced by aldosterone may represent an alternative etiology for vascular diseases such as hypertension . 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 . Differentiation in vivo of classical non-steroidal antiinflammatory drugs from cytokine suppressive antiinflammatory drugs and other pharmacological classes using mouse tumour necrosis factor alpha production . The stimulation of tumour necrosis factor alpha ( P01375 alpha ) production by lipopolysaccharide ( LPS ) has been widely used , both in vitro and in vivo , to examine the biochemistry and pharmacology of inflammatory cytokine production . It appears that classical nonsteroidal antiinflammatory drugs ( NSAIDs ) ( prostaglandin H synthase 1 ( P23219 ) inhibitors ) do not inhibit but instead stimulate cytokine production . In the current study , the authors utilized LPS-induced P01375 alpha production in the Balb/c mouse to evaluate the activity of a classical NSAID , a mixed inhibitor , and SmithKline Beecham cytokine suppressive antiinflammatory drugs ( CSAID ) . The results corroborated the stimulation of P01375 alpha production by NSAIDs ( indomethacin , naproxen , ibuprofen ) and indicated that the stimulation rank-ordered with the potency of inhibition of P23219 . Neither acetaminophen nor nabumetone was found to stimulate P01375 alpha production significantly . Tenidap , a compound reported to inhibit P09917 , cyclooxygenase and cytokine production , also stimulated P01375 alpha production while the P09917 inhibitor , phenidone , was inactive . The CSAID ( exemplified by SK & F 86002 , SK & F 105809 and SK & F 104351 ) , strongly inhibited P01375 alpha production in this model system ( ED50s of 32 , 48 , and 34 mg/kg p.o. , respectively ) . These results clearly differentiate CSAID from the other compounds tested and suggest that CSAID are relatively weak inhibitors of PGHS 1 while being potent inhibitors of inflammatory cytokine production . Enhancement of 5-fluorouracil efficacy on high P35354 expressing HCA-7 cells by low dose indomethacin and NS-398 but not on low P35354 expressing HT-29 cells . The antiproliferative effect of 5-fluorouracil ( DB00544 ) in the presence of low dose non-steroidal anti-inflammatory drugs ( NSAIDs ) on high cyclooxygenase-2 ( P35354 ) -expressing HCA-7 and low P35354 -expressing HT-29 colon carcinoma cell lines was investigated . Pharmacogenetic parameters were studied to characterize the DB00544 sensitivity of the two cell lines . P04818 ( TS ) and methylenetetrahydrofolate reductase ( P42898 ) polymorphisms were determined by PCR analysis . Cell proliferation was measured by P50991 assay , cell cycle distribution and apoptosis by FACS analysis . Cyclooxygenase expression was detected by Western blot and also by fluorescence microscopy . Prostaglandin E(2) ( PGE(2) ) levels were investigated with ELISA kit . The HT-29 cell line was found to be homozygous for TS 2R and 1494ins6 and T homozygous for P42898 677 polymorphisms predicting high DB00544 sensitivity ( IC(50) : 10 microM ) . TS 3R homozygosity , TS 1496del6 and P42898 677CT heterozygosity may explain the modest DB00544 sensitivity ( IC(50) : 1.1 mM ) of the HCA-7 cell line . Indomethacin and NS-398 ( 10 microM and 1.77 microM , respectively ) reduced the PGE(2) level in HCA-7 cells ( > 90 % ) . Low concentrations of NSAIDs without antiproliferative potency increased the S-phase arrest and enhanced the cytotoxic action of DB00544 only in HCA-7 cells after 48-hours treatment . The presented data suggested that the enhancement of DB00544 cytotoxicity by indomethacin or NS-398 applied in low dose is related to the potency of NSAIDs to modulate the cell-cycle distribution and the apoptosis ; however , it seems that this effect might be dependent on cell phenotype , namely on the P35354 expression . 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 . DB00998 Vernalis . Vanguard ( now Vernalis ) has developed frovatriptan , a selective P28222 /1D partial agonist licensed from GlaxoSmithKline as a potential treatment for migraine [ 188478 ] , [ 194382 ] , [ 377863 ] . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape .

MH_train_1421

MH_train_1421

MH_train_1421

interacts_with DB04868?

multiple_choice

Transcriptional regulation of the cannabinoid receptor type 1 gene in T cells by cannabinoids . Effects of cannabinoids ( CBs ) are mediated by two types of receptors , P21554 and CB2 . In this report , we investigated whether CBs regulate gene expression of their cognate receptors in T cells and studied underlying mechanisms in P01730 + Jurkat T cells . Transcription of the P21554 gene was strongly induced in response to Delta9-tetrahydrocannabinol ( THC ) , whereas the CB2 gene was not regulated . The induction of P21554 gene expression is mediated by CB2 receptors only , as demonstrated by using the P21554 and CB2 agonists R(+)-methanandamide and JWH 015 , respectively , and combinations of THC plus P21554 - and CB2-specific antagonists . After activation of CB2 receptors , the transcription factor P42229 is phosphorylated . P42229 then transactivates P05112 . Induction of P05112 mRNA as well as P05112 protein release from the cells are necessary for the following induction of the P21554 gene . This was demonstrated by using decoy oligonucleotides against P42229 , which blocked P05112 and P21554 mRNA induction , and by using the P05112 receptor antagonist P05112 [ R121D,Y124D ] , which blocked the up-regulation of P21554 gene transcription . Transactivation of the P21554 gene in response to P05112 is then mediated by the transcription factor P42226 , as shown by using decoy oligonucleotides against P42226 . An increase in P21554 -mediated phosphorylation of MAPK in cells prestimulated with CB2-specific agonists suggests up-regulation of functional P21554 receptor proteins . In summary , up-regulation of P21554 in T lymphocytes in response to CBs themselves may facilitate or enhance the various immunomodulatory effects related to CBs . 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 . Serious obstetric complications interact with hypoxia-regulated/vascular-expression genes to influence schizophrenia risk . The etiology of schizophrenia is thought to include both epistasis and gene-environment interactions . We sought to test whether a set of schizophrenia candidate genes regulated by hypoxia or involved in vascular function in the brain ( P31749 , P23560 , O75052 , P36544 , P21964 , Q96EV8 , Q99259 , Q14832 , Q99466 , Q02297 , O43272 , P49798 , P01375 ) interacted with serious obstetric complications to influence risk for schizophrenia . A family-based study of transmission disequilibrium was conducted in 116 trios . Twenty-nine probands had at least one serious obstetric complication ( OC ) using the McNeil-Sjostrom Scale , and many of the OCs reported were associated with the potential for fetal hypoxia . Analyses were conducted using conditional logistic regression and a likelihood ratio test ( LRT ) between nested models was performed to assess significance . Of the 13 genes examined , four ( P31749 ( three SNPs ) , P23560 ( two SNPs ) , Q96EV8 ( one SNP ) and Q14832 ( one SNP ) ) showed significant evidence for gene-by-environment interaction ( LRT P-values ranged from 0.011 to 0.037 ) . Although our sample size was modest and the power to detect interactions was limited , we report significant evidence for genes involved in neurovascular function or regulated by hypoxia interacting with the presence of serious obstetric complications to increase risk for schizophrenia . 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 . [ 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 . Presynaptic serotonergic inhibition of GABAergic synaptic transmission in mechanically dissociated rat basolateral amygdala neurons . 1. The basolateral amygdala ( P00519 ) nuclei contribute to the process of anxiety . GABAergic transmission is critical in these nuclei and serotonergic inputs from dorsal raphe nuclei also significantly regulate GABA release . In mechanically dissociated rat P00519 neurons , spontaneous miniature inhibitory postsynaptic currents ( mIPSCs ) arising from attached GABAergic presynaptic nerve terminals were recorded with the nystatin-perforated patch method and pharmacological isolation . 2 . 5-HT reversibly reduced the GABAergic mIPSC frequency without affecting the mean amplitude . The serotonergic effect was mimicked by the P08908 specific agonist 8-OH DPAT ( 8-hydroxy-2-(di-n-propylamino)tetralin ) and blocked by the P08908 antagonist spiperone . 3 . The GTP-binding protein inhibitor N-ethylmaleimide removed the serotonergic inhibition of mIPSC frequency . In either K+-free or Ca2+-free external solution , 5-HT could inhibit mIPSC frequency . 4 . High K+ stimulation increased mIPSC frequency and 8-OH DPAT inhibited this increase even in the presence of Cd2+ . 5 . DB02587 , an activator of adenylyl cyclase ( AC ) , significantly increased synaptic GABA release frequency . Pretreatment with forskolin prevented the serotonergic inhibition of mIPSC frequency in both the standard and high K+ external solution . 6 . Ruthenium Red ( RR ) , an agent facilitating the secretory process in a Ca2+-independent manner , increased synaptic GABA release . 5-HT also suppressed RR-facilitated mIPSC frequency . 7 . We conclude that 5-HT inhibits GABAergic mIPSCs by inactivating the AC- DB02527 signal transduction pathway via a G-protein-coupled P08908 receptor and this intracellular pathway directly acts on the GABA-releasing process independent of K+ and Ca2+ channels in the presynaptic nerve terminals . Determination of ancestral allele for possible human cancer-associated polymorphisms . To determine ancestral allele in possible cancer-associated polymorphisms , DNA samples from 10 chimpanzees ( Pan troglodytes ) were sequenced for alleles corresponding to 17 polymorphisms : 8 short tandem repeats [ P18510 ( alias IL-1RA ) variable number tandem repeat ( VNTR ) ; P04818 ( previously TS ) VNTR ; AR CAG repeat ; dinucleotide repeats of P22309 , IGF1 , P01579 ( alias P01579 ) , P03372 ( alias P03372 ) , and P00533 ] and 9 single nucleotide polymorphisms ( P03956 -1607 1G/2G , P08254 -1171 5A/6A , O15527 Ser326Cys , P05091 Gly487Lys , P04637 Arg72Pro , Q9UNQ0 Gln141Lys , P16455 Leu84Phe , P04179 Ala-9Val , and P42898 Ala222Val ) . No chimpanzee polymorphism corresponded to human P18510 VNTR ; the ancestral allele was a repeat lost in humans . Dinucleotide repeat polymorphisms of IGF1 , P01579 , P03372 , and P00533 were shared by chimpanzees , but the length of repeats tended to be longer in humans than in chimpanzees . This tendency was particularly evident for IGF1 . All of the SNPs tested are human-specific nucleotide changes . The ancestral allele 7A was shown to be lost in P08254 -1171 5A/6A . Thus , all of the possible cancer-associated polymorphisms tested have human-specific alleles , and the ancestral allele is lost in three polymorphisms ( P18510 VNTR , P22309 CA repeat , and P08254 -1171 5A/6A ) , suggesting a possible involvement of human-specific alleles in cancer susceptibility . Inhibition of rat liver UDP-glucuronosyltransferase by silymarin and the metabolite silibinin-glucuronide . The inhibitory effects of silymarin , its main constituent silibinin and the metabolite silibinin-glucuronide on UDP-glucuronosiltransferase ( P78381 ) were evaluated in rat hepatic microsomes . Three substrates were chosen to cover both P22309 and UGT2B family isozymes : bilirubin ( substrate of P22309 ) , p-nitrophenol ( P19224 ) and ethinylestradiol ( UGT2B1 and 2B3 for position C17 and P22309 for position P01024 ) . The study of p-nitrophenol and bilirubin glucuronidation indicated that silymarin ( SM ) and silibinin glucuronide ( SB-G ) were enzyme inhibitors . The kinetic analysis showed that the type of inhibition was competitive in all cases and the Ki obtained were : for p-nitrophenol glucuronidation , KiSB-Gapp : 14+/-1 microg/ml and KiSMapp : 51+/-10 microg/ml and for bilirubin glucuronidation , KiSB-Gapp : 16+/-3 microg/ml . In turn , ethinylestradiol glucuronidation was not affected by any of the compounds studied suggesting that the inhibitory effect was restricted to P22309 isozymes . Similar studies performed using human hepatic microsomes showed that SM and SB-G were also inhibitors of human P22309 isozymes . In conclusion , administration of silymarin or its main constituent silibinin could lead to the decrease in the glucuronidation of substrates whose conjugation depends on P22309 isozymes in a process mediated by silibinin-glucuronide , though their effect in humans needs further investigation . Causation , prevention and reversal of vascular endothelial dysfunction . P01308 Resistance along with endothelial dysfunction give rise to a constellation of syndromes designated as P41252 / O14974 metabolic syndrome . Endothelial dysfunction starts early in life much before the development of structural atherosclerosis . Recent insights into vascular biology enable us to understand the molecular mechanisms underlying endothelial dysfunction , and the scope and need for prevention of " pre-clinical " coronary atherosclerosis through lifestyle modification ; diet , exercise and stress management . Diminished production of nitric oxide ( NO ) and/or increased inactivation of NO through oxidative stress ( reactive oxygen species ROS and reactive nitrogen species ( RNS ) are the basis of endothelial dysfunction hence increasing the bioavailability of NO and decreasing its inactivation is the aim of prevention and reversal of endothelial dysfunction . P01308 regulates constitutive NOS gene expression in endothelial cells in vivo ; vasodilation is an important component of P01308 -stimulated whole body glucose uptake . Successful strategies are : Q07869 alpha and gamma agonists which increase NO production in endothelium ; anti-oxidants such as vit . E and C ; supplementation with L-arginine , tetrahydrobiopterin-BH4 or sepiapterin ( precursor of BH4 ) , SOD mimetic tempol , statins which apart from lowering cholesterol improve NO production , selective beta1 adrenoreceptor antagonists such as nebivolol ; suppression of angiotensin-mediated endothelin production by P12821 inhibitors and ATR blockers ; P21554 receptor blockers , PKCb inhibitors , nitric oxide donors ( glyceryl trinitrate and isosorbide dinitrate ) , dietary supplements of EPA/ DB01708 and regular physical exercise and control of mental stress . Novel real-time polymerase chain reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia . The recent World Health Organization classification recognizes different subtypes of acute myeloid leukemia ( AML ) according to the presence of several recurrent genetic abnormalities . Detection of these abnormalities and other molecular changes is of increasing interest because it contributes to a refined diagnosis and prognostic assessment in AML and enables monitoring of minimal residual disease . These genetic abnormalities can be detected using single RT-PCR , although the screening is still labor intensive and costly . We have developed a novel real-time RT-PCR assay to simultaneously detect 15 AML-associated rearrangements that is a simple and easily applicable method for use in clinical diagnostic laboratories . This method showed 100 % specificity and sensitivity ( 95 % confidence interval , 91 % to 100 % and 92 % to 100 % , respectively ) . The procedure was validated in a series of 105 patients with AML . The method confirmed all translocations detected using standard cytogenetics and fluorescence in situ hybridization and some additional undetected rearrangements . Two patients demonstrated two molecular rearrangements simultaneously , with P11274 - P00519 implicated in both , in addition to Q01196 -MECOM in one patient and P29590 - P10276 in another . In conclusion , this novel real-time RT-PCR assay for simultaneous detection of multiple AML-associated fusion genes is a versatile and sensitive method for reliable screening of recurrent rearrangements in AML . Regulation of pregnane X receptor ( O75469 ) function and P22309 gene expression by posttranslational modification of O75469 protein . Human UDP-glucuronosyltransferase ( P78381 ) 1A1 is a critical enzyme responsible for detoxification and metabolism of endogenous and exogenous lipophilic compounds such as bilirubin . The present study shows how cyclin-dependent kinase ( CDK ) inhibitor roscovitine stimulated the expression of P22309 in HepG2 cells . O75469 ( O75469 ) -mediated transactivation of P22309 reporter gene was more prominently enhanced by roscovitine , compared with the basal- , constitutive androstane receptor ( CAR ) - , and aryl hydrocarbon receptor-mediated activities . We determined the regulatory mechanism of P22309 expression through O75469 's stimulation by roscovitine . Although phosphomimetic mutations at Thr290 and Thr408 retained the O75469 protein in cytoplasm and attenuated the induction of P22309 expression by both roscovitine and rifampicin , a mutation at Ser350 specifically reduced the activity of O75469 induced by roscovitine . Immunoprecipitation analysis revealed that the T290D but not T408D mutant protein remained in cytoplasm by forming a complex with heat shock protein 90 and cytoplasmic CAR retention protein , whereas treatment with proteasome inhibitor MG-132 accumulated the T408D mutant protein in cytoplasm . Transfection with anti- P24941 small interfering RNA ( siRNA ) but not anti- P06493 or Q00535 siRNA led to enhanced expression of P22309 . S350D yellow fluorescent protein- O75469 fusion protein could translocate from cytoplasm to nucleus similar to the wild-type protein but was detected as an acetylated protein , whose binding with retinoid X receptor ( RXR ) and histone deacetylase was impaired . Cotransfection with coactivator steroid receptor coactivator ( P12931 ) 2 but not Q15788 partly recovered its O75469 activity . These results indicate that roscovitine stimulated the expression of P22309 by inhibiting P24941 , which phosphorylated O75469 at Ser350 to suppress binding with RXR and coactivator and maintain the acetylation of O75469 protein . 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 . [ Concentration of acute phase proteins in serum of children with rheumatoid arthritis ] . An attempts was made at evaluation of the changes of acute phase proteins-seromucoid ( BRS ) , alpha 1-acid glycoprotein ( alpha 1-AGP ) , alpha 1-antitrypsin ( alpha 1-AT ) , haptoglobin ( Hp ) , P01024 protein of complement system ( P01024 ) , P05155 ( DB05341 ) , and transferrin ( Tf ) in the serum of children with rheumatoid arthritis . The patients were divided into clinical and age groups . The studies carried out have shown in ill children increase of BRS , alpha 1-AGP , alpha 1-AT , Hp , and DB05341 levels , During remission increased levels of alpha 1-AGP and Hp persisted . In the studied clinical groups a positive correlation was found between the intensity of changes of the studied indices and the degree of disease activity . In the age groups greater increase of the levels of the studied acute phase proteins was found in the group of preschool children and the group of puberty spurt . The obtained results suggest that the determination of acute phase proteins may be useful in laboratory investigation of children with rheumatoid arthritis . 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 . CB2 receptors in the brain : role in central immune function . Recently , it has been recognized that the cannabinoid receptor CB2 may play a functionally relevant role in the central nervous system ( CNS ) . This role is mediated primarily through microglia , a resident population of cells in the CNS that is morphologically , phenotypically , and functionally related to macrophages . These cells also express the cannabinoid receptor P21554 . The P21554 receptor ( CB1R ) is constitutively expressed at low levels while the CB2 receptor ( CB2R ) is expressed at higher levels and is modulated in relation to cell activation state . The relatively high levels of the CB2R correspond with microglia being in ' responsive ' and ' primed ' states , suggesting the existence of a ' window ' of functional relevance during which activation of the CB2R modulates microglial activities . Signature activities of ' responsive ' and ' primed ' microglia are chemotaxis and antigen processing , respectively . The endocannabinoid 2-arachidonylglycerol has been reported to stimulate a chemotactic response from these cells through the CB2R . In contrast , we have shown in vivo and in vitro that the exogenous cannabinoids DB00470 and CP55940 inhibit the chemotactic response of microglia to Acanthamoeba culbertsoni , an opportunistic pathogen that is the causative agent of Granulomatous Amoebic Encephalitis , through activation of the CB2R . It is postulated that these exogenous cannabinoids superimpose an inhibitory effect on pro-chemotactic endocannabinoids that are elicited in response to Acanthamoeba . Furthermore , the collective results suggest that the CB2R plays a critical immune functional role in the CNS . 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 . 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 . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS ) P01116 mutations : analytical considerations . Colorectal cancer ( CRC ) is the third most common cancer and the second most common cause of cancer death globally . Significant improvements in survival have been made in patients with metastasis by new therapies . For example , Cetuximab and DB01269 are monoclonal antibodies that inhibit the epidermal growth receptor ( P00533 ) . P01116 mutations in codon 12 and 13 are the recognized biomarkers that are analyzed in clinics before the administration of anti- P00533 therapy . Genetic analyses have revealed that mutations in P01116 predict a lack of response to DB01269 and Cetuximab in patients with metastatic CRC ( mCRC ) . Notably , it is estimated that 35-45 % of CRC patients harbor P01116 mutations . Therefore , P01116 mutation testing should be performed in all individuals with the advanced CRC in order to identify the patients who will not respond to the monoclonal P00533 antibody inhibitors . New techniques for P01116 testing have arisen rapidly , and each technique has advantages and disadvantages . Herein , we review the latest published literature specific to P01116 mutation testing techniques . Since reliability and feasibility are important issues in clinical analyses . Therefore , this review also summarizes the effectiveness and limitations of numerous P01116 mutation testing techniques . The O60656 enzyme is a peroxisome proliferator-activated receptor alpha and gamma target gene . Peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma are ligand-activated transcription factors belonging to the nuclear receptor family . Q07869 alpha mediates the hypolipidemic action of the fibrates , whereas Q07869 gamma is a receptor for the antidiabetic glitazones . In the present study , the UDP-glucuronosyltransferase ( P78381 ) 1A9 enzyme is identified as a Q07869 alpha and Q07869 gamma target gene . UGTs catalyze the glucuronidation reaction , which is a major pathway in the catabolism and elimination of numerous endo- and xenobiotics . Among the P22309 family enzymes , O60656 metabolizes endogenous compounds , including catecholestrogens , and xenobiotics , such as fibrates and to a lesser extent troglitazone . Treatment of human hepatocytes and macrophages and murine adipocytes with activators of Q07869 alpha or Q07869 gamma resulted in an enhanced O60656 expression and activity . In addition , disruption of the Q07869 alpha gene in mice completely abolished the Q07869 alpha agonist-induced O60656 mRNA and activity levels . A Q07869 response element was identified in the promoter of O60656 at positions -719 to -706 bp by transient transfection and electromobility shift assays . Considering the role of O60656 in catecholestrogen metabolism , Q07869 alpha and Q07869 gamma activation may contribute to the protection against genotoxic catecholestrogens by stimulating their inactivation in glucuronide derivatives . Furthermore , since O60656 is involved in the catabolism of fibrates , these results suggest that Q07869 alpha and Q07869 gamma may control the intracellular level of active fibrates . Activation of P28335 /2 receptors depresses polysynaptic reflexes and excitatory amino acid-induced motoneuron responses in frog spinal cord . DB02772 gap recordings from the ventral roots of isolated , hemisected frog spinal cords were used to evaluate the effects of high concentrations of serotonin ( 5-HT ) and alpha-methyl-5-HT ( alpha-Me-5-HT ) on the changes in motoneuron potential produced by dorsal root stimulation and by excitatory amino acids and agonists . Bath application of 5-HT in concentrations of 10 microM or greater produced a concentration-dependent motoneuron depolarization . Polysynaptic ventral root potentials evoked by dorsal root stimuli were reduced in both amplitude and area by 5-HT or alpha-Me-5-HT ( both 100 microM ) . This may result from a reduction of the postsynaptic sensitivity of motoneurons to excitatory amino acid transmitters because 5-HT significantly depressed motoneuron depolarizations produced by addition of L-glutamate and L-aspartate to the superfusate . Similarly , 5-HT reduced depolarizations produced by the excitatory amino acid agonists N-methyl-D-aspartate ( DB01221 ) , quisqualate , alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid ( AMPA ) , and kainate . alpha-Me-5-HT reduced DB01221 depolarizations . DB05232 ( TTX ) did not affect the ability of 5-HT to attenuate DB01221 or kainate depolarizations , but did eliminate the 5-HT-induced attenuation of quisqualate and AMPA depolarizations . The glycine receptor site associated with the DB01221 receptor did not appear to be affected by 5-HT because saturation of the site by excess glycine did not alter the 5-HT-induced depression of DB01221 responses . The P28335 /2 antagonist ketanserin and the P08908 /2 antagonist spiperone significantly attenuated the 5-HT-induced depression of DB01221 -depolarizations. ( ABSTRACT TRUNCATED AT 250 WORDS )

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P01236 influence on cytosol and nuclear androgen receptors in the ventral , dorsal , and lateral lobes of the rat prostate . PRL augments testosterone-mediated growth of the prostate in a permissive manner . To elucidate the mechanism of this hormonal interaction , the present study examined the effect of PRL on cytosol and nuclear androgen receptors in the three prostate lobes . Adult male Sprague-Dawley rats were castrated , given 5-mm Silastic implants of testosterone , and either grafted with two anterior pituitary glands under the kidney capsule or sham operated . Three weeks later , animals were killed , serum was collected for PRL and testosterone RIA , and the ventral , dorsal , and lateral prostate lobes were processed for either cytosol or nuclear androgen receptor quantitation . Pituitary grafts significantly elevated the serum PRL concentration and increased the weight and the content of protein and DNA of the lateral prostate lobe compared to control values . There was no effect on these parameters in the ventral or dorsal lobes . P10275 levels and apparent distributions were different in the ventral , dorsal , and lateral lobes of control animals . Unoccupied and total cytosolic androgen receptors in the three separate prostate lobes were not significantly affected by the presence of the grafts . However , an elevated PRL concentration was associated with an increase ( P less than 0.005 ) in nuclear androgen receptor content in the lateral lobe exclusively . The binding affinity was not altered by pituitary grafts in any of the lobes . These findings suggest that PRL promotes lateral prostatic growth by increasing nuclear androgen receptor levels in that tissue and , thus , optimizes its response to circulating testosterone . 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 . Molecular basis of differential resistance to cycloguanil and pyrimethamine in Plasmodium falciparum malaria . DB01131 and pyrimethamine are antifolate drugs with distinct chemical structures that are used commonly in the prophylaxis and treatment of Plasmodium falciparum malaria . Clinical reports and field studies have suggested that some parasites refractory to proguanil can be treated with pyrimethamine , and vice versa . Analysis of the P. falciparum dihydrofolate reductase ( P00374 ) from different parasites reveals the structural basis for differential susceptibility to these antifolate drugs . Parasites harboring a pair of point mutations from Ala-16 to DB00161 -16 and from DB00133 -108 to DB00156 -108 are resistant to cycloguanil ( the active metabolite of proguanil ) but not to pyrimethamine . A single DB00174 -108 mutation , on the other hand , confers resistance to pyrimethamine with only a moderate decrease in susceptibility to cycloguanil . Significant cross-resistance to both drugs occurs in parasites having mutations that include DB00133 -108 ---- DB00174 -108 and DB00167 -164 ---- DB00149 -164 . These results reflect the distinct structures of pyrimethamine and cycloguanil and suggest fine differences in binding within the active site cavity of P00374 . Alternative inhibitors , used alone or in combination , may be effective against some strains of cycloguanil- or pyrimethamine-resistant malaria . 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 . Genetics of Cushing 's syndrome . Cushing 's syndrome ( CS ) is characterized by pathologically elevated free glucocorticoid levels . Endogenous hypercortisolism is usually due to DB01285 -secreting pituitary corticotropic adenomas and less often due to ectopic DB01285 -secreting neuroendocrine neoplasms or DB01285 -independent adrenal cortisol hypersecretion . CS is a serious chronic disease leading to a several-fold increase in cardiovascular morbidity and mortality . Multiple genetic alterations have been described in the setting of sporadic corticotropinoma formation . Changes in the expression profiles have been demonstrated in growth factors and their receptors , cell-cycle regulators and in various genes related to hormonal gene transcription , synthesis and secretion . Sporadic adrenal adenomas and carcinomas may demonstrate dysfunction in genes such as P04637 among others . Cushing 's disease can be an inherited condition also . Multiple endocrine neoplasia type 1 ( O00255 ) and familial isolated pituitary adenomas ( FIPA ) together account for 5 % of pituitary adenomas . Cushing 's disease occurs infrequently in an inherited setting in both of these conditions . To date only 2 cases of Cushing 's disease have been described in association with mutations in AIP . One case of Cushing 's disease has been reported as part of MEN4 , a rare O00255 -like syndrome due to mutation in the P46527 gene . Carney complex ( CNC ) due to P10644 mutations in most cases is associated with CS , mainly as a cause of bilateral adrenal hyperplasia . The DB02527 signaling pathway is affected in this setting . In recent times the involvement of genes such as Q9HCR9 , O95263 and others have expanded the spectrum of the genetic pathophysiology of CS . 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 . Effects of the dopamine D3 receptor ( P35462 ) gene polymorphisms on risperidone response : a pharmacogenetic study . Previous observations of the anatomical distribution and pharmacological profile of the dopamine D(3) receptor ( P35462 ) have indicated its potential role in antipsychotic drug action . DB00734 , an effective first-line atypical antipsychotic agent , exhibits a relatively high affinity for this receptor . Recent studies have reported an association of the Ser9Gly polymorphism in the P35462 gene with therapeutic response to risperidone , but the results were inconsistent . We therefore postulated that the Ser9Gly polymorphism might be in linkage disequilibrium with an undetected variant that exerts a direct influence on risperidone efficacy . The present study genotyped eight single nucleotide polymorphisms ( SNPs ) distributed throughout the P35462 gene and examined five of these for association with treatment outcome , following an 8-week period of risperidone monotherapy in 130 schizophrenic patients from mainland China . Clinical symptoms were assessed before and after the treatment period , using the Brief Psychiatry Rating Scale ( BPRS ) . The confounding effects of non-genetic factors were estimated and the baseline symptom score was included as a covariate for adjustment . Neither was any association observed between the five polymorphisms and improvement in total BPRS scores nor was any combined effect of these variants detected in the haplotype analysis . The current results indicate that genetic variations within the P35462 gene may not contribute significantly to interindividual differences in the therapeutic efficacy of risperidone . Genetic pathway-based hierarchical clustering analysis of older adults with cognitive complaints and amnestic mild cognitive impairment using clinical and neuroimaging phenotypes . Hierarchical clustering is frequently used for grouping results in expression or haplotype analyses . These methods can elucidate patterns between measures that can then be applied to discerning their validity in discriminating between experimental conditions . Here a hierarchical clustering method is used to analyze the results of an imaging genetics study using multiple brain morphology and cognitive testing endpoints for older adults with amnestic mild cognitive impairment ( D6RGH6 ) or cognitive complaints ( CC ) compared to healthy controls ( HC ) . The single nucleotide polymorphisms ( SNPs ) are a subset of those included on a larger array that are found in a reported Alzheimer 's disease ( AD ) and neurodegeneration pathway . The results indicate that genetic models within the endpoints cluster together , while there are 4 distinct sets of SNPs that differentiate between the endpoints , with most significant results associated with morphology endpoints rather than cognitive testing of patients ' reported symptoms . The genes found in at least one cluster are P08183 , Q02410 , P56817 , Q9Y5Z0 , P10415 , Q07817 , P55210 , P28329 , P01034 , P35462 , P21918 , P05231 , Q07954 , NAT1 , and P49810 . The greater associations with morphology endpoints suggests that changes in brain structure can be influenced by an individual 's genetic background in the absence of dementia and in some cases ( Cognitive Complaints group ) even without those effects necessarily being detectable on commonly used clinical tests of cognition . The results are consistent with polygenic influences on early neurodegenerative changes and demonstrate the effectiveness of hierarchical clustering in identifying genetic associations among multiple related phenotypic endpoints . 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 . 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 . 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 . 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 . 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 . Roles of E3 ubiquitin ligases in cell adhesion and migration . Recent studies have demonstrated that a number of E3 ubiquitin ligases , including Cbl , Smurf1 , Smurf2 , HDM2 , BCA2 , P21583 (beta-TRCP) and XRNF185 , play important roles in cell adhesion and migration . Cbl negatively regulates cell adhesion via alpha integrin and Rap1 and inhibits actin polymerization by ubiquitinating mDab1 and Q9Y6W5 . Smurf1 regulates cell migration through ubiquitination of RhoA , talin head domain and hPEM2 , while Smurf2 ubiquitinates Smurf1 , TGFbeta type I receptor and RaplB to modulate cell migration and adhesion . HDM2 negatively regulates cell migration by targeting NFAT ( a transcription factor ) for ubiquitination and degradation , while P21583 (beta-TRCP) ubiquitinates Snail ( a transcriptional repressor of P12830 ) to inhibit cell migration . Q13049 promotes cell migration through ubiquitination of Q9NYB9 ( Abi2 ) , a tumor suppressor . Q99942 and XRNF185 modulate cell migration by ubiquitinating paxillin . Thus , these E3 ubiquitin ligases regulate cell adhesion and ( or ) migration through ubiquitination of their specific substrates . 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 . 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 . [ Association analysis of the essential hypertension susceptibility genes in adolescents : Kangwha study ] . OBJECTIVES : In this study we examined the association between the genetic markers P12821 ( A-240T , C-93T , I/D , A2350G ) , AGT ( M235T ) , AT1R ( A1166C ) , P19099 ( T-344C , V386A ) , REN ( G2646A ) , P07550 ( G46A , C79G , T-47C , T164I ) , P16520 ( C825T ) and P35611 ( G460W ) and the presence of essential hypertension in adolescents . METHODS : The Kangwha Study is an 18-year prospective study that is aimed at elucidating the determinants of the blood pressure level from childhood to early adulthood . For this study , we constructed a case-control dataset of size of 277 and 40 family trios data from the Kangwha Study . For this purpose , we perform a single locus-based case-control association study and a single locus-based P04053 ( transmission/disequilibrium test ) study . RESULTS : In the case-control study , the single locus-based association study indicated that the P35611 ( G460W ) ( p = 0.0403 ) , AGT ( M235T ) ( p = 0.0002 ) , and REN ( G2646A ) ( p = 0.0101 ) markers were significantly associated with the risk of hypertension . These results were not confirmed on the P04053 study . This study showed that genetic polymorphisms of the P35611 , AGT and REN genes might be related to the hypertension in Korean adolescents . CONCLUSIONS : This study provided useful information on genetics markers related to blood pressure . Further study will be needed to confirm the effect of the alpha adducin gene , the angiotensinogen gene and the renin gene on essential hypertension . 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 . 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 . E3 ubiquitin ligase Q13049 negatively regulates tumor suppressor p53 to promote tumorigenesis . P04637 has a key role in maintaining genomic stability and preventing tumorigenesis through its regulation of cellular stress responses , including apoptosis , cell cycle arrest and senescence . To ensure its proper levels and functions in cells , p53 is tightly regulated mainly through post-translational modifications , such as ubiquitination . Here , we identified E3 ubiquitin ligase Q13049 as a novel p53 target gene and negative regulator to regulate p53-mediated stress responses . In response to stress , such as DNA damage , p53 binds to the p53 responsive element in the promoter of the Q13049 gene and transcriptionally induces the expression of Q13049 in cells . In turn , Q13049 interacts with p53 and promotes p53 degradation through ubiquitination . Thus , Q13049 negatively regulates p53-mediated apoptosis , cell cycle arrest and senescence in response to stress . Q13049 is frequently overexpressed in different types of human tumors . Q13049 overexpression promotes cell oncogenic transformation and tumorigenesis in mice in a largely p53-dependent manner . Taken together , our results demonstrated that as a novel p53 target and a novel negative regulator for p53 , Q13049 has an important role in regulation of p53 and p53-mediated cellular stress responses . Furthermore , our results also revealed that impairing p53 function is a novel mechanism for Q13049 in tumorigenesis . 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 . Antagonism of Q9GZP0 by human antibody DB05139 prevents renal scarring in experimental glomerulonephritis . Glomerular mesangial cell proliferation and/or matrix accumulation characterizes many progressive renal diseases . Q9GZP0 was identified recently as a novel mediator of mesangial cell proliferation in vitro and in vivo . This study investigated the long-term consequences of Q9GZP0 inhibition in vivo . Rats with progressive mesangioproliferative glomerulonephritis ( uninephrectomy plus anti-Thy-1.1 antibody ) received the Q9GZP0 -neutralizing , fully human mAb DB05139 on days 3 , 10 , and 17 after disease induction . Glomerular mesangioproliferative changes on day 10 were significantly reduced by anti- Q9GZP0 treatment as compared with control antibody . Eight weeks after disease induction , anti- Q9GZP0 therapy significantly ameliorated focal segmental glomerulosclerosis , podocyte damage ( de novo desmin expression ) , tubulointerstitial damage , and fibrosis as well as the accumulation of renal interstitial matrix including type III collagen and fibronectin . Treatment with anti- Q9GZP0 also reduced the cortical infiltration of monocytes/macrophages on day 56 , possibly related to lower renal cortical complement activation ( C5b-9 deposition ) and/or reduced epithelial-to-mesenchymal transition ( preserved cortical expression of P12830 and reduced expression of vimentin and alpha-smooth muscle actin ) . In conclusion , these data provide evidence for a causal role of Q9GZP0 in the pathogenesis of renal scarring and point to a new therapeutic approach to progressive mesangioproliferative renal disease .

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interacts_with DB00015?

multiple_choice

[ Molecular mechanism of cardiovascular damage induced by aldosterone ] . Although the pro-inflammatory and pro-fibrotic actions of aldosterone on the vasculature have been reported , the effects and molecular mechanisms of aldosterone on endothelial function are yet to be determined . We investigated how aldosterone regulates endothelial nitric oxide synthase ( P29474 ) function in human umbilical vein endothelial cells ( HUVECs ) . HUVECs were incubated for 16 hrs with 10(-7) mol/l of aldosterone . The concentration of reactive oxygen species ( ROS ) was estimated by measuring DCF chemiluminescence . Signal transduction was estimated by Western immunoblots . Realtime RT-PCR was performed to measure expression of transcripts of endogenous GTP cyclohydrolase-1 ( P30793 ) and components of NAD(P)H oxidase . In order to eliminate the possible effect of the glucocorticoid receptor ( GR ) , and to emphasize the role of mineralocorticoid receptor ( MR ) , we used GR siRNA and knocked down GR expression in several experiments . NO output was estimated by intracellular cGMP concentration . ROS production increased significantly in aldosterone-treated HUVEC , but was abolished by pre-treatment with eplerenone . Transcripts of p47(phox) were increased by aldosterone treatment . Vascular endothelial growth factor ( P15692 ) -induced P29474 DB00133 1177 but not Akt DB00133 473 phosphorylation levels were reduced significantly by pretreatment with aldosterone . Pretreatment with either eplerenone or okadaic acid restored phosphorylation levels of P29474 DB00133 1177 in aldosterone-treated cells , suggesting that protein phosphatase ( PP ) 2A was upregulated by aldosterone via MR . The decrease in NO output caused by aldosterone pretreatment was reversed significantly by either DB00360 ( BH(4) ) , P30793 overexpression , or p47(phox) knockdown . These results suggest that aldosterone inhibits P29474 function through bimodal mechanisms of BH(4) deficiency and PP2A activation . A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 and P35354 ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg . Pancreatic gene expression during the initiation of acute pancreatitis : identification of P18146 as a key regulator . We hypothesized that genes expressed in pancreatic acinar cells during the initiation of acute pancreatitis determine the severity of the disease . Therefore , we utilized microarrays to identify those genes commonly induced in rat pancreatic acinar cells within 1-4 h in two in vivo models , caerulein and taurocholate administration . This strategy yielded 51 known genes representing a complex array of molecules , including those that are likely to either reduce or increase the severity of the disease . Novel genes identified in the current study included P18847 , BRF1 , C/EBPbeta , P80511 , P18146 , ephrinA1 , villin2 , ferredoxin , latexin , lipocalin , P28562 , NGFI-B , RhoA , tissue factor ( TF ) , and syndecan . To validate these microarray results , the role of P18146 was further investigated using quantitative RT-PCR , Western blotting , and immunocytochemistry . P18146 expression occurred within acinar cells and correlated with the development of caerulein-induced acute pancreatitis in rats . Furthermore , the levels of the inflammation-related genes P13500 , P05121 , TF , P05231 , and P05362 and the extent of lung inflammation were reduced during the initiation of caerulein-induced acute pancreatitis in P18146 -deficient mice . Thus this study identified P18146 and several other novel genes likely to be important in the development and severity of acute pancreatitis . [ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity . [ 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 . Endothelial histamine H1 receptor signaling reduces blood-brain barrier permeability and susceptibility to autoimmune encephalomyelitis . Disruption of the blood-brain barrier ( BBB ) underlies the development of experimental autoimmune encephalomyelitis ( EAE ) and multiple sclerosis . Environmental factors , such as Bordetella pertussis , are thought to sensitize central endothelium to biogenic amines like histamine , thereby leading to increased BBB permeability . B. pertussis-induced histamine sensitization ( Bphs ) is a monogenic intermediate phenotype of EAE controlled by histamine H(1) receptor ( Hrh1/H(1)R ) . Here , we transgenically overexpressed H(1)R in endothelial cells of Hrh1-KO ( H(1)RKO ) mice to test the role of endothelial H(1)R directly in Bphs and EAE . Unexpectedly , transgenic H(1)RKO mice expressing endothelial H(1)R under control of the P04275 promoter ( H(1)RKO- P04275 ( P35367 ) Tg ) were Bphs-resistant . Moreover , H(1)RKO- P04275 ( P35367 ) Tg mice exhibited decreased BBB permeability and enhanced protection from EAE compared with H(1)RKO mice . Thus , contrary to prevailing assumptions , our results show that endothelial H(1)R expression reduces BBB permeability , suggesting that endothelial H(1)R signaling may be important in the maintenance of cerebrovascular integrity . 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 . 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 . Tandutinib ( MLN518/CT53518 ) targeted to stem-like cells by inhibiting the function of DB00171 -binding cassette subfamily G member 2 . Tandutinib is a novel inhibitor of tyrosine kinases P36888 , P09619 and P10721 . Our study was to explore the capability of DB05465 to reverse ABC transporter-mediated multidrug resistance . Tandutinib reversed Q9UNQ0 -mediated drug resistance in Q9UNQ0 -482-R2 , Q9UNQ0 -482-G2 , Q9UNQ0 -482-T7 and S1-M1-80 cells and increased the accumulation of doxorubicin , rhodamine 123 and [ H(3) ] mitoxantrone in Q9UNQ0 -overexpressing cells . Importantly , DB05465 selectively sensitized side population cells to mitoxantrone . Taken together , our results advocate the potency of DB05465 as an Q9UNQ0 modulator and stem-like cells targeted agent to increase efficiency of anticancer drugs . P23560 and basic fibroblast growth factor downregulate DB01221 receptor function in cerebellar granule cells . Evidence has accumulated to suggest that the DB01221 glutamate receptor subtype plays an important role in neuronal degeneration evoked by hypoxia , ischemia , or trauma . Cerebellar granule cells in culture are vulnerable to DB01221 -induced neuronal excitotoxicity . In these cells , brain-derived neurotrophic factor ( P23560 ) and basic fibroblast growth factor ( P09038 ) prevent the excitotoxic effect of DB01221 . However , little is known about the molecular mechanisms underlying the protective properties of these trophic factors . Using cultured rat cerebellar granule cells , we investigated whether P23560 and P09038 prevent DB01221 toxicity by downregulating DB01221 receptor function . Western blot and RNase protection analyses were used to determine the expression of the various DB01221 receptor subunits ( Q9UHB4 , Q12879 , Q13224 , and Q14957 ) after P23560 or P09038 treatment . P09038 and P23560 elicited a time-dependent decrease in the expression of Q12879 and Q14957 subunits . Because DB01221 receptor activation leads to increased intracellular Ca2+ concentration ( [Ca2+]i ) , we studied the effect of the P23560 - and P09038 -induced reduction in Q12879 and Q14957 synthesis on the DB01221 -evoked Ca2+ responses by single-cell fura-2 fluorescence ratio imaging . P23560 and P09038 reduced the DB01221 -mediated [Ca2+]i increase with a time dependency that correlates with their ability to decrease Q12879 and Q14957 subunit expression , suggesting that these trophic factors also induce a functional downregulation of the DB01221 receptor . Because sustained [Ca2+]i is believed to be causally related to neuronal injury , we suggest that P23560 and P09038 may protect cerebellar granule cells against excitotoxicity by altering the DB01221 receptor-Ca2+ signaling via a downregulation of DB01221 receptor subunit expression . Functional and molecular characterization of ex vivo cultured epiretinal membrane cells from human proliferative diabetic retinopathy . Characterization of the cell surface marker phenotype of ex vivo cultured cells growing out of human fibrovascular epiretinal membranes ( fvERMs ) from proliferative diabetic retinopathy ( PDR ) can give insight into their function in immunity , angiogenesis , and retinal detachment . FvERMs from uneventful vitrectomies due to PDR were cultured adherently ex vivo . Surface marker analysis , release of immunity- and angiogenesis-pathway-related factors upon P01375 α activation and measurement of the intracellular calcium dynamics upon mechano-stimulation using fluorescent dye Fura-2 were all performed . FvERMs formed proliferating cell monolayers when cultured ex vivo , which were negative for endothelial cell markers ( CD31 , P35968 ) , partially positive for hematopoietic- ( P28906 , Q08722 ) and mesenchymal stem cell markers ( CD73 , CD90/Thy-1 , and P09619 β ) , and negative for CD105 . CD146/ P43121 and CD166/ Q13740 , previously unreported in cells from fvERMs , were also expressed . Secretion of 11 angiogenesis-related factors ( DPPIV/ P27487 , P58294 / P30613 , ET-1 , P18065 and 3 , P10145 / P10145 , P13500 / P13500 , P14780 , PTX3/ P26022 , P05121 / P05121 , P36955 / P36955 , P01033 , and P07996 -1 ) were detected upon P01375 α activation of fvERM cells . Mechano-stimulation of these cells induced intracellular calcium propagation representing functional viability and role of these cells in tractional retinal detachment , thus serving as a model for studying tractional forces present in fvERMs in PDR ex vivo . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . 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 . 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 . 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 . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . 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 . DB06699 : a novel gonadotropin-releasing hormone blocker for the treatment of prostate cancer . Androgen deprivation therapy with gonadotropin releasing-hormone ( DB00644 ) receptor agonists provides the mainstay of endocrine treatment for advanced prostate cancer . Although effective , DB00644 agonists induce an initial testosterone surge , which can cause painful and potentially dangerous clinical flare . DB06699 is a novel P30968 blocker that provides immediate , profound and sustained testosterone reduction , without an initial surge . In a Phase III trial , degarelix and leuprolide showed similar long-term efficacy in maintaining testosterone levels of 0.5 ng/ml or less over 1 year , and induced significantly faster testosterone and prostate-specific antigen suppression . DB06699 was well tolerated ; the most common side effects were mild/moderate injection-site reactions and hot flashes . Findings to date suggest that degarelix may make an important contribution to the treatment of prostate cancer .

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interacts_with DB04908?

multiple_choice

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 . 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 . The role of heat shock proteins in Amyotrophic Lateral Sclerosis : The therapeutic potential of DB05025 . DB05025 is a hydroxylamine derivative , a group of compounds which have unique properties as co-inducers of heat shock protein expression , but only under conditions of cellular stress . DB05025 has been found to be neuroprotective in a number of neurodegenerative disease models , including Amyotrophic Lateral Sclerosis ( P35858 ) , and in mutant Superoxide Dismutase 1 ( P00441 ) mice that model P35858 , DB05025 rescues motor neurons , improves neuromuscular function and extends lifespan . The therapeutic potential of DB05025 is currently under investigation in a Phase II clinical trial for P35858 patients with P00441 mutations . In this review we summarize the evidence for the neuroprotective effects of enhanced heat shock protein expression by DB05025 and other inducers of the Heat Shock Response . P35858 is a complex , multifactorial disease affecting a number of cell types and intracellular pathways . Cells and pathways affected by P35858 pathology and which may be targeted by a heat shock protein-based therapy are also discussed in this review . For example , protein aggregation is a characteristic pathological feature of neurodegenerative diseases including P35858 . Enhanced heat shock protein expression not only affects protein aggregation directly , but can also lead to more effective clearance of protein aggregates via the unfolded protein response , the proteasome-ubiquitin system or by autophagy . However , compounds such as DB05025 have effects beyond targeting protein mis-handling and can also affect additional pathological mechanisms such as oxidative stress . Therefore , by targeting multiple pathological mechanisms , compounds such as DB05025 may be particularly effective in the development of a disease-modifying therapy for P35858 and other neurodegenerative disorders . 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 . 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 . DB00886 , a dual angiotensin-converting enzyme and neutral endopeptidase inhibitor , prevents fatty streak deposit in apolipoprotein E-deficient mice . P12821 ( P12821 ) is mainly responsible for converting angiotensin I ( AI ) to angiotensin II ( AII ) , and P12821 inhibitors prevent atherosclerosis in animal models . P08473 ( NEP ) degrades DB05875 , kinins and atrial natriuretic peptide ( P01160 ) , and aortic wall NEP activity was found to be increased in atherosclerosis . In the present study , we have evaluated the effect of candoxatril , a NEP inhibitor , and of omapatrilat , a dual P12821 and NEP inhibitor , on the development of fatty streak in apolipoprotein E ( apoE ) -deficient mice . Groups of ten male apoE-deficient mice were given either placebo , candoxatril 50 mg/kg per day , or omapatrilat 10 , or 100 mg/kg per day for 4 months . None of the treatments influenced body weight , serum total or HDL-cholesterol . Compared with the placebo , candoxatril did not protect the mice from fatty streak deposit . In contrast , omapatrilat dose dependently inhibited the constitution of fatty streak in apoE-deficient mice . The precise advantages of the dual P12821 and NEP inhibition versus the inhibition of only P12821 should now be considered in the prevention of atherosclerosis as well as in the occurrence of its complications . 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 . 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 . Vascular endothelial growth factor ( P15692 ) inhibition by small molecules . Angiogenesis is essential for primary tumours to grow and metastasise , and is driven by the production of positive angiogenic factors . The Vascular Endothelial Growth Factor ( P15692 ) family is central to the process of angiogenesis and comprises 5 molecules designated A , B , C , D and E. P15692 is overexpressed in several solid malignancies . The actions of P15692 are mediated through receptors possessing tyrosine kinase activity : P17948 ( Flt-1 ) , P35968 ( Kdr/Flk-1 ) and P35916 ( Flt-4 ) . Anti- P15692 strategies include the use of antibodies to P15692 or its receptors , the use of ribozymes to decrease receptor expression , and the use of inhibitors of tyrosine kinase to reduce receptor activation and downstream signalling . The focus of this review is small molecule inhibitors of P15692 receptors which target their intrinsic tyrosine kinase activity . The clinical development of the following agents is discussed : SU5416 , SU11248 , SU6668 , DB04879 , DB05294 . Stimulation of cloned human serotonin P28221 beta receptor sites in stably transfected P13671 glial cells promotes cell growth . The involvement of serotonin P28221 beta receptor sites was investigated in the growth of rat P13671 glial cells permanently transfected with a gene encoding a human P28221 beta receptor . The 5-HT receptor identity of control and transfected P13671 glial/ P28221 beta cells was determined by reverse transcription-polymerase chain reaction using primers specific for rat P08908 , rat P28222 , rat P28221 alpha , human P28221 beta , and rat 5- Q13049 receptor genes . Constitutive mRNA for 5- Q13049 receptors was present in control and transfected P13671 glial/ P28221 beta cells , whereas mRNA for P28221 beta receptor sites was only present in the transfected P13671 glial/ P28221 beta cell line . 5-HT inhibited forskolin-stimulated cyclic AMP formation and promoted cell growth , in contrast to the absence of any measurable effect in pcDNA3 plasmid-transfected and nontransfected P13671 glial cells . The 5-HT effects could be mimicked by sumatriptan ( EC50 = 44-76 nM ) and were totally and partially blocked by methiothepin ( IC50 = 9 nM ) and GR 127,935 ( 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol-3-yl)-biphenyl-4-carbox yli c acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide ; IC50 = 97 pM ) , respectively . No effect on cell growth was measured with the 5-HT2 receptor agonist DOI [ 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ; 10 microM ] , suggesting that 5- Q13049 receptors are not involved in the 5-HT-stimulated P13671 glial/ P28221 beta cell growth . Dibutyryl-cyclic AMP ( 0.3 mM ) -treated cultures did not show sumatriptan-promoted cell growth , indicating an inhibitory role for cyclic AMP in the cell growth mediated by P28221 beta receptor sites. ( ABSTRACT TRUNCATED AT 250 WORDS ) Serotonin and fluoxetine modulate bone cell function in vitro . Recent studies have proposed a role for serotonin and its transporter in regulation of bone cell function . In the present study , we examined the in vitro effects of serotonin and the serotonin transporter inhibitor fluoxetine " DB00472 " on osteoblasts and osteoclasts . Human mononuclear cells were differentiated into osteoclasts in the presence of serotonin or fluoxetine . Both compounds affected the total number of differentiated osteoclasts as well as bone resorption in a bell-shaped manner . RT-PCR on the human osteoclasts demonstrated several serotonin receptors , the serotonin transporter , and the rate-limiting enzyme in serotonin synthesis , tryptophan hydroxylase 1 ( Tph1 ) . Tph1 expression was also found in murine osteoblasts and osteoclasts , indicating an ability to produce serotonin . In murine pre-osteoclasts ( RAW264.7 ) , serotonin as well as fluoxetine affected proliferation and NFkappaB activity in a biphasic manner . Proliferation of human DB05914 ( O60682 ) and primary osteoblasts ( NHO ) , and 5- Q13049 receptor expression was enhanced by serotonin . DB00472 stimulated proliferation of O60682 and murine preosteoblasts ( MC3T3-E1 ) in nM concentrations , microM concentrations were inhibitory . The effect of fluoxetine seemed direct , probably through 5-HT2 receptors . Serotonin-induced proliferation of MC3T3-E1 cells was inhibited by the PKC inhibitor ( GF109203 ) and was also markedly reduced when antagonists of the serotonin receptors P41595 /C or 5- Q13049 /C were added . Serotonin increased osteoprotegerin ( O00300 ) and decreased receptor activator of NF-kappaB ligand ( O14788 ) secretion from osteoblasts , suggesting a role in osteoblast-induced inhibition of osteoclast differentiation , whereas fluoxetine had the opposite effect . This study further describes possible mechanisms by which serotonin and the serotonin transporter can affect bone cell function . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . 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 . Alterations in receptors for thyrotropin-releasing hormone , serotonin , and acetylcholine in amyotrophic lateral sclerosis . We utilized quantitative autoradiography to examine thyrotropin-releasing hormone ( TRH ) receptors , serotonin type 1A ( P08908 ) receptors , muscarinic cholinergic receptors , choline uptake sites , beta-adrenergic receptors , and norepinephrine uptake sites in discrete laminae of spinal cord from patients with amyotrophic lateral sclerosis ( P35858 ) and non-neurologic controls . We found decreases of over 50 % in the concentration of TRH receptors in lamina IX of cervical , thoracic , and lumbar spinal cord from P35858 patients . Similar reductions were noted in concentrations of muscarinic cholinergic receptors in lamina IX of spinal cords from P35858 patients . Significant increases of up to 140 % in P08908 receptor densities were noted in lamina IX of spinal cords from P35858 patients . No differences were noted between the concentrations of beta-adrenergic receptors or norepinephrine uptake sites in patients with P35858 and controls . These findings suggest that TRH and 5-HT may be involved in the pathophysiology of P35858 , and act in a comodulatory role in the normal spinal cord . 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 . A cyclophilin A inducible expressed in gonad of zhikong scallop Chlamys farreri . P62937 ( CypA ) , a receptor for the immunosuppressive agent cyclosporin A ( DB00091 ) , is a cis-trans peptidyl-prolyl isomerase ( PPIase ) which accelerates the cis-trans isomerization of prolyl-peptide bonds , interacts with a variety of proteins and therefore regulates their activities . One CypA ( designated CfCypA ) cDNA was cloned from Chlamys farreri by expressed sequence tag ( EST ) and rapid amplification of cDNA ends ( RACE ) techniques . The full-length cDNA of CfCypA consisted of 1,248 nucleotides with a canonical polyadenylation signal sequence AATAAA , a poly ( A ) tail , and an open reading frame ( ORF ) of 495 nucleotides encoding a polypeptide of 164 amino acids . The deduced amino acid sequence shared high similarity with CypA from the other species , indicating that CfCypA should be a new member of the CypA family . Quantitative real-time ( RT ) PCR was employed to assess the mRNA expression of CfCypA in various tissues and its temporal expression in haemocytes and gonad of scallops challenged with Vibrio anguillarum . The mRNA transcripts of CfCypA could be detected in all the examined tissues with highest expression level in gonad . After bacterial challenge , the expression level of CfCypA was almost unchanged in haemocytes , but up-regulated in gonad and increased to the peak ( 22.59-fold ; P < 0.05 ) at 4 h post-injection , and then dropped to the original level at 8 h post-injection . These results indicated that CfCypA was constitutive expressed in haemocytes , but could be induced in gonad , and perhaps played a critical role in response to the bacterial challenge in gonad . Direct and irreversible inhibition of cyclooxygenase-1 by nitroaspirin ( DB05822 ) . Benzoic acid , 2-(acetyl-oxy)-3-[(nitrooxy)methyl]phenyl ester ( DB05822 ) , a new drug made by an aspirin molecule linked , through a spacer , to a nitric oxide ( NO ) -donating moiety , is now under clinical testing for the treatment of atherothrombotic conditions . DB00945 exerts its antithrombotic activity by irreversibly inactivating platelet cyclooxygenase ( P36551 ) -1 . DB05822 in vivo undergoes metabolism into deacetylated and/or denitrated metabolites , and it is not known whether DB05822 needs to liberate aspirin to inhibit P23219 , or whether it can block it as a whole molecule . The aim of our study was to evaluate the effects of DB05822 and its analog or metabolites on platelet P23219 and whole blood P35354 and on purified ovine P36551 ( oCOX ) -1 and oCOX-2 . In particular , we have compared the mechanism by which DB05822 inhibits purified oCOX enzymes with that of aspirin using a spectrophotometric assay . All the DB05822 derivatives containing acetylsalicylic acid inhibited the activity of oCOX-1 and oCOX-2 , whereas the deacetylated metabolites and the nitric oxide-donating moiety were inactive . Dialysis experiments showed that oCOX-1 inhibition by DB05822 , similar to aspirin , is irreversible . Reversible P36551 inhibitors ( indomethacin ) or salicylic acid incubated with the enzyme before DB05822 prevent the irreversible inhibition of oCOX-1 by DB05822 as well as by aspirin . In conclusion , our data show that DB05822 acts as a direct and irreversible inhibitor of P23219 and that the presence of a spacer and NO-donating moiety in the molecule slows the kinetics of P23219 inhibition by DB05822 , compared with aspirin . 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 . Effect of glial cell line-derived neurotrophic factor on behavior and key members of the brain serotonin system in mouse strains genetically predisposed to behavioral disorders . The effect of glial cell line-derived neurotrophic factor ( P39905 ) on behavior and on the serotonin ( 5-HT ) system of a mouse strain predisposed to depressive-like behavior , ASC/Icg ( Antidepressant Sensitive Cataleptics ) , in comparison with the parental " nondepressive " CBA/Lac mice was studied . Within 7 days after acute administration , P39905 ( 800 ng , i.c.v. ) decreased cataleptic immobility but increased depressive-like behavioral traits in both investigated mouse strains and produced anxiolytic effects in ASC mice . The expression of the gene encoding the key enzyme for 5-HT biosynthesis in the brain , tryptophan hydroxylase-2 ( Tph-2 ) , and P08908 receptor gene in the midbrain as well as 5- Q13049 receptor gene in the frontal cortex were increased in P39905 -treated ASC mice . At the same time , P39905 decreased P08908 and 5- Q13049 receptor gene expression in the hippocampus of ASC mice . P39905 failed to change Tph2 , P08908 , or 5- Q13049 receptor mRNA levels in CBA mice as well as 5-HT transporter gene expression and P08908 and 5- Q13049 receptor functional activity in both investigated mouse strains . The results show 1 ) a P39905 -induced increase in the expression of key genes of the brain 5-HT system , Tph2 , P08908 , and 5- Q13049 receptors , and 2 ) significant genotype-dependent differences in the 5-HT system response to P39905 treatment . The data suggest that genetically defined cross-talk between neurotrophic factors and the brain 5-HT system underlies the variability in behavioral response to P39905 . DB01599 alleviates atherosclerosis and improves high density lipoprotein function . BACKGROUND : DB01599 is a unique hypolipidemic agent that decreases high density lipoprotein cholesterol ( HDL-C ) . However , it is not definite that whether probucol hinders the progression of atherosclerosis by improving HDL function . METHODS : Eighteen New Zealand White rabbits were randomly divided into the control , atherosclerosis and probucol groups . Control group were fed a regular diet ; the atherosclerosis group received a high fat diet , and the probucol group received the high fat diet plus probucol . Hepatocytes and peritoneal macrophages were isolated for [ (3)H ] labeled cholesterol efflux rates and expression of O95477 and SR-B1 at gene and protein levels ; venous blood was collected for serum paraoxonase 1 , myeloperoxidase activity and lipid analysis . Aorta were prepared for morphologic and immunohistochemical analysis after 12 weeks . RESULTS : Compared to the atherosclerosis group , the paraoxonase 1 activity , cholesterol efflux rates , expression of O95477 and Q8WTV0 in hepatocytes and peritoneal macrophages , and the level of O95477 and Q8WTV0 in aortic lesions were remarkably improved in the probucol group , But the serum HDL cholesterol concentration , myeloperoxidase activity , the IMT and the percentage plaque area of aorta were significantly decreased . CONCLUSION : DB01599 alleviated atherosclerosis by improving HDL function . The mechanisms include accelerating the process of reverse cholesterol transport , improving the anti-inflammatory and anti-oxidant functions .

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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 . Cytoprotective-selective activated protein C attenuates Pseudomonas aeruginosa-induced lung injury in mice . Inhibition of the small GTPase RhoA attenuates the development of pulmonary edema and restores positive alveolar fluid clearance in a murine model of Pseudomonas aeruginosa pneumonia . DB00055 ( aPC ) blocks the development of an unfavorably low ratio of small GTPase Rac1/RhoA activity in lung endothelium through endothelial protein C receptor ( Q9UNN8 ) /protease-activated receptor-1 ( P25116 ) -dependent signaling mechanisms that include transactivating the sphingosine-1-phosphate ( Q14703 ) pathway . However , whether aPC 's cytoprotective effects can attenuate the development of pulmonary edema and death associated with P. aeruginosa pneumonia in mice remains unknown . Thus , we determined whether the normalization of a depressed ratio of activated Rac1/RhoA by aPC would attenuate the P. aeruginosa-mediated increase in protein permeability across lung endothelial and alveolar epithelial barriers . Pretreatment with aPC significantly reduced P. aeruginosa-induced increases in paracellular permeability across pulmonary endothelial cell and alveolar epithelial monolayers via an inhibition of RhoA activation and a promotion of Rac1 activation that required the Q9UNN8 - P25116 and Q14703 pathways . Furthermore , pretreatment with aPC attenuated the development of pulmonary edema in a murine model of P. aeruginosa pneumonia . Finally , a cytoprotective-selective aPC mutant , aPC-5A , which lacks most of aPC 's anticoagulant activity , reproduced the protective effect of wild-type aPC by attenuating the development of pulmonary edema and decreasing mortality in a murine model of P. aeruginosa pneumonia . Taken together , these results demonstrate a critical role for the cytoprotective activities of aPC in attenuating P. aeruginosa-induced lung vascular permeability and mortality , suggesting that cytoprotective-selective aPC-5A with diminished bleeding risks could attenuate the lung damage caused by P. aeruginosa in critically ill patients . 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 . P01308 -like growth factor-1 potentiates platelet activation via the P41252 / P42336 pathway . As insulin-like growth factor-1 ( DB01277 ) is present in the alpha granules of platelets and its receptor is expressed on the platelet surface , it may contribute to the amplification of platelet responses and pathogenesis of cardiovascular disease . The functional and signaling pathways that are involved in DB01277 modulation of platelet function , however , are presently unknown . Here , I report that DB01277 stimulation of platelets results in dose-dependent phosphorylation of the IGF receptor in the range of 1 to 100 nM . Phosphorylation of the IGF receptor is rapid and sustained , with maximal phosphorylation reached within 1 minute . Furthermore , DB01277 stimulates tyrosine phosphorylation of insulin receptor substrate-1 ( P35568 ) and Q9Y4H2 and their association with the p85 subunit of phosphoinositide-3 kinase ( PI3K ) . DB01277 -stimulated tyrosine phosphorylation of P35568 and Q9Y4H2 and subsequent p85 binding is transient and precedes phosphorylation of protein kinase B ( P31749 ) on Ser473 . P25116 -mediated platelet aggregation is potentiated by DB01277 and this potentiation , together with P31749 phosphorylation , is abolished by the P42336 inhibitors PI-103 and PIK-75 . Importantly , the IGF receptor inhibitor DB00238 -AEW541 and the neutralization antibody alphaIR3 inhibit SFLLRN-stimulated aggregation , implicating DB01277 in autocrine regulation of platelet function . These results demonstrate that DB01277 activates the IGF receptor/ P41252 /PI3K/ P31749 pathway , and that P42336 is essential for the potentiatory effect of DB01277 on platelet responses . Retinoid-binding proteins in craniofacial development . Cephalic neural crest cells are known to form the frontonasal mesenchyme and contribute to the mesenchyme of the visceral arches . Retinoids affect neural crest cells and their derivatives during development , and thus cause craniofacial , thymus , and conotruncal heart malformations . In addition , retinoids induce malformations of the central nervous system ( CNS ) . Retinoic acid ( RA ) and its congeners accumulate in a saturable manner in neural crest and neural crest-derived cells , in the hindbrain , and the spinal cord of mouse embryos . Cellular retinoic acid-binding protein ( CRABP ) was localized by immunohistochemistry in the same areas as were the labelled RA congeners . Thus , CRABP and RA congeners were found in the transitional zone between surface ectoderm and neuropeithelium , from where neural crest cells are known to emanate ( day 8 1/2 ) . Later , specific labelling was found in the frontonasal mesenchyme and in the visceral arches . Also in the trunk , neural crest cells were labelled . In CNS , strong staining was seen in the rhombomeres ( especially numbers 4-6 ) of the hindbrain and in the spinal cord . DB00162 and cellular retinol-binding protein ( P09455 ) were more evenly distributed , with exception of surface ectoderm , epithelium of gut , and myocardium , where P09455 was specifically expressed . These findings are discussed in relation to the differential expression of nuclear RA receptors and homeobox genes in the craniofacial region and in the hindbrain . It is possible that RA is important for the normal pattern formation in these regions and acts as a morphogen as previously proposed in limb development . 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 . Mechanism of the induction of the differentiation of HL-60 leukemia cells by antifolates . The classic inhibitor of dihydrofolate reductase ( P00374 ) , methotrexate ( MTX ) , has been shown to be an effective inducer of the differentiation of HL-60 promyelocytic leukemia cells ( Bodner A.J. et al. ; J. Natl. Cancer Inst. 67:1025-1030 ; 1981 ) . We have obtained evidence that induction of the differentiation of these cells by MTX , as well as by other folic acid antagonists , is the result of the effects of these agents on purine and thymine nucleotide biosynthesis . DB04485 ( 10 microM ) completely blocked both the cytotoxicity and induction of differentiation produced by the specific inhibitor of thymidylate synthase ( TS ) , N10-propargyl-5,8-dideazafolic acid ( CB-3717 ) . DB04485 also blocked the acute cytotoxicity caused by MTX and trimetrexate ( DB01157 ) ; the induction of differentiation and the loss of proliferative capacity , however , were only partially prevented by thymidine . DB04076 ( 100 microM ) , which completely restored antifolate-depleted purine nucleotide levels , had no effect on either the cytotoxicity or the induction of maturation produced by these agents . The growth inhibitory effects and the induction of differentiation caused by dideazatetrahydrofolic acid ( DDATHF ) , which acts on de novo purine nucleotide biosynthesis rather than on P00374 or TS , was completely prevented by hypoxanthine . DB04076 also completely prevented the inhibition of cellular replication and induction of differentiation by MTX and DB01157 when combined with thymidine . The findings suggest that the depletion of intracellular thymine nucleotide levels by the antifolates , MTX , DB01157 , and CB-3717 is the primary event involved in the maturation of HL-60 leukemia cells produced by these agents and that maturation occurs concomitantly with a high level of cytotoxicity. ( ABSTRACT TRUNCATED AT 250 WORDS ) A structural and in vitro characterization of asoprisnil : a selective progesterone receptor modulator . Selective progesterone receptor modulators ( SPRMs ) have been suggested as therapeutic agents for treatment of gynecological disorders . One such SPRM , asoprisnil , was recently in clinical trials for treatment of uterine fibroids and endometriosis . We present the crystal structures of progesterone receptor ( PR ) ligand binding domain complexed with asoprisnil and the corepressors nuclear receptor corepressor ( NCoR ) and Q9Y618 . This is the first report of steroid nuclear receptor crystal structures with ligand and corepressors . These structures show PR in a different conformation than PR complexed with progesterone ( P4 ) . We profiled asoprisnil in PR-dependent assays to understand further the PR-mediated mechanism of action . We confirmed previous findings that asoprisnil demonstrated antagonism , but not agonism , in a PR-B transfection assay and the T47D breast cancer cell alkaline phosphatase activity assay . Asoprisnil , but not DB00834 , weakly recruited the coactivators Q15788 and Q9Y6Q9 . However , asoprisnil strongly recruited the corepressor NCoR in a manner similar to DB00834 . Unlike DB00834 , NCoR binding to asoprisnil-bound PR could be displaced with equal affinity by NCoR or Q15596 peptides . We further showed that it weakly activated T47D cell gene expression of Sgk-1 and O60437 and antagonized P4-induced expression of both genes . In rat leiomyoma ELT3 cells , asoprisnil demonstrated partial P4-like inhibition of cyclooxygenase ( P36551 ) enzymatic activity and P35354 gene expression . In the rat uterotrophic assay , asoprisnil demonstrated no P4-like ability to oppose estrogen . Our data suggest that asoprisnil differentially recruits coactivators and corepressors compared to DB00834 or P4 , and this specific cofactor interaction profile is apparently insufficient to oppose estrogenic activity in rat uterus . P12004 D-cdk4 activity modulates the subnuclear localization and interaction of Q02078 with P12931 -family coactivators during skeletal muscle differentiation . Prior work has indicated that D-type cyclin-cdk4 complexes , which are only active in proliferating cells , can suppress the skeletal muscle differentiation program in proliferating myoblasts . In this study , we show that cyclin D-cdk activity can block the activity of the Q02078 family of transcriptional regulators , which are crucial regulators of skeletal muscle gene expression . We have found that cyclin D-cdk activity blocks the association of Q06413 with the coactivator protein Q9Y3R0 and thereby inhibits the activity of Q02078 . During skeletal muscle differentiation , Q9Y3R0 is localized to punctate nuclear structures and can apparently tether Q02078 to such structures . Cotransfection of Q9Y3R0 can both potentiate the transcriptional activity of a Gal4- Q06413 construct and induce Q06413 localization to punctate nuclear structures . Consistent with the absence of punctate nuclear Q9Y3R0 in proliferating myoblasts , we have found that ectopic cyclin D-cdk4 expression disrupts the localization of both Q9Y3R0 and Q06413 to these punctate subnuclear structures . Our findings indicate that cyclin D-cdk4 activity represses skeletal muscle differentiation in proliferating cells by blocking the association of Q02078 with the coactivator Q9Y3R0 and concomitantly disrupts the association of these factors with punctate nuclear subdomains within the cell . 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 . 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 . 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) . Functional gene expression profile underlying methotrexate-induced senescence in human colon cancer cells . Cellular functions accompanying establishment of premature senescence in methotrexate-treated human colon cancer C85 cells are deciphered in the present study from validated competitive expression microarray data , analyzed with the use of Ingenuity Pathways Analysis ( IPA ) software . The nitrosative/oxidative stress , inferred from upregulated expression of inducible nitric oxide synthase ( P35228 ) and mitochondrial dysfunction-associated genes , including monoamine oxidases P21397 and P27338 , β-amyloid precursor protein ( P05067 ) and presenilin 1 ( P49768 ) , is identified as the main determinant of signaling pathways operating during senescence establishment . Activation of p53-signaling pathway is found associated with both apoptotic and autophagic components contributing to this process . Activation of nuclear factor κB ( NF-κB ) , resulting from interferon γ ( IFNγ ) , integrin , interleukin 1β ( IL-1β ) , P05112 , P35225 , Q9GZX6 , Toll-like receptors ( TLRs ) 1 , 2 and 3 , growth factors and tumor necrosis factor ( P01375 ) superfamily members signaling , is found to underpin inflammatory properties of senescent C85 cells . Upregulation of P38936 -activated kinases ( Q13177 and Q9NQU5 ) , several Rho molecules and myosin regulatory light chains P19105 and O14950 , indicates acquisition of motility by those cells . Mitogen-activated protein kinase p38 MAPK β , extracellular signal-regulated kinases P28482 and Q13164 , protein kinase B P31749 , as well as calcium , are identified as factors coordinating signaling pathways in senescent C85 cells . DB00563 induces apoptosis through p53/ P38936 -dependent pathway and increases P12830 expression through downregulation of HDAC/ Q15910 . DB00563 ( MTX ) is a dihydrofolate reductase ( P00374 ) inhibitor widely used as an anticancer drug in different kinds of human cancers . Here we investigated the anti-tumor mechanism of MTX against non-small cell lung cancer ( NSCLC ) A549 cells . MTX not only inhibited in vitro cell growth via induction of apoptosis , but also inhibited tumor formation in animal xenograft model . RNase protection assay ( RPA ) and RT-PCR demonstrated its induction of p53 target genes including DR5 , P38936 , Puma and Noxa . Moreover , MTX promoted p53 phosphorylation at Ser15 and acetylaion at Lys373/382 , which increase its stability and expression . The apoptosis and inhibition of cell viability induced by MTX were dependent on p53 and , partially , on P38936 . In addition , MTX also increased P12830 expression through inhibition of histone deacetylase ( HDAC ) activity and downregulation of polycomb group protein enhancer of zeste homologue 2 ( Q15910 ) . Therefore , the anticancer mechanism of MTX acts through initiation of p53-dependent apoptosis and restoration of P12830 expression by downregulation of HDAC/ Q15910 . 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 . Exendin-4 stimulates islet cell replication via the IGF1 receptor activation of mTORC1/ P23443 . P43220 ( P43220 ) agonists , such as exendin-4 , potentiate glucose-stimulated insulin secretion and are currently used in the management of type 2 diabetes . Interestingly , P43220 agonists also have the ability to augment β-cell mass . In this report , we provide evidence that in the presence of glucose , exendin-4 stimulates rodent islet cell DNA replication via the activation of ribosomal protein S6 kinase 1 ( P23443 ) and that this is mediated by the protein kinase B ( P31749 ) -dependent activation of P42345 complex 1 ( mTORC1 ) . We show that activation of this pathway is caused by the autocrine or paracrine activation of the IGF1 receptor ( P08069 ) , as siRNA-mediated knockdown of the P08069 effectively blocked exendin-4-stimulated P31749 and mTORC1 activation . In contrast , pharmacological inactivation of the epidermal growth factor receptor has no discernible effect on exendin-4-stimulated P31749 or mTORC1 activation . Therefore , we conclude that P43220 agonists stimulate β-cell proliferation via the P31749 -dependent stimulation of mTORC1/ P23443 whose activation is mediated through the autocrine/paracrine activation of the P08069 . This work provides a better understanding of the molecular basis of GLP1 agonist-induced β-cell proliferation which could potentially be exploited in the identification of novel drug targets that increase β-cell mass . The transcriptional coactivator DRIP/mediator complex is involved in vitamin D receptor function and regulates keratinocyte proliferation and differentiation . Mediator is a multisubunit coactivator complex that facilitates transcription of nuclear receptors . We investigated the role of the mediator complex as a coactivator for vitamin D receptor ( P11473 ) in keratinocytes . Using P11473 affinity beads , the vitamin D receptor interacting protein ( DRIP ) /mediator complex was purified from primary keratinocytes , and its subunit composition was determined by mass spectrometry . The complex included core subunits , such as Q15648 /MED1 ( MED1 ) , that directly binds to P11473 . Additional subunits were identified that are components of the RNA polymerase II complex . The functions of different mediator components were investigated by silencing its subunits . The core subunit MED1 facilitates P11473 activity and regulating keratinocyte proliferation and differentiation . A newly described subunit Q13503 also has a role in promoting keratinocyte proliferation and differentiation , whereas Q9BTT4 has an inhibitory role . Blocking MED1/ Q13503 expression caused hyperproliferation of keratinocytes , accompanied by increases in mRNA expression of the cell cycle regulator cyclin D1 and/or glioma-associated oncogene homolog . Blocking MED1 or Q13503 expression also resulted in defects in calcium-induced keratinocyte differentiation , as indicated by decreased expression of differentiation markers and decreased translocation of P12830 to the membrane . These results show that keratinocytes use the transcriptional coactivator mediator to regulate P11473 functions and control keratinocyte proliferation and differentiation . P09455 -1 expression in ovarian cancer : a potential therapeutic target . BACKGROUND/AIM : Cellular retinol binding protein-1 regulates retinol bioavailability and contributes to cell differentiation maintenance , but its role in ovarian carcinogenesis remains uncertain . We investigated P09455 -1 expression in ovarian tumors and P09455 -1 signaling-regulated pathways . MATERIALS AND METHODS : We performed immunohistochemistry , methylation-specific PCR , gene copy number analysis in ovarian tumors and proliferation/apoptosis evaluation , gene array , blot and real-time PCR in P09455 -1-transfected A2780 ovarian cancer cells . RESULTS : P09455 -1 expression was reduced or absent in G2 and P46379 ovarian carcinomas . P09455 -1 silencing in 60 % of G2 and 66.7 % of P46379 carcinomas was due to P09455 -1 promoter methylation . A2780 P09455 -1-transfected cells showed increased retinol-induced apoptosis , retinoid-induced reduced clonogenicity and down-regulation of proliferation and transcription genes , including P31749 , Q9Y243 , P00533 , P01100 , P05412 , P42224 and P42229 . CONCLUSION : P09455 -1 loss in G2/ P46379 ovarian carcinomas and increased apoptotic susceptibility to retinoids in P09455 -1-transfected-A2780 cells suggest P09455 -1 screening as a target to ensure efficacy of an adjuvant retinoid therapy . 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 . 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 . 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 . miR-21 overexpression enhances TGF-β1-induced epithelial-to-mesenchymal transition by target smad7 and aggravates renal damage in diabetic nephropathy . Epithelial-to-mesenchymal transition ( EMT ) plays an important role in renal interstitial fibrosis ( Q9HBH0 ) with diabetic nephropathy ( DN ) . O15105 ( a inhibitory smad ) , a downstream signaling molecules of TGF-β1 , represses the EMT . The physiological function of miR-21 is closely linked to EMT and Q9HBH0 . However , it remained unclear whether miR-21 over-expression affected TGF-β1-induced EMT by regulating smad7 in DN . In this study , real-time RT-PCR , cell transfection , luciferase reporter gene assays , western blot and confocal microscope were used , respectively . Here , we found that miR-21 expression was upregulated by TGF-β1 in time- and concentration -dependent manner . Moreover , miR-21 over-expression enhanced TGF-β1-induced EMT ( upregulation of a-SMA and downregulation of P12830 ) by directly down-regulating smad7/p-smad7 and indirectly up-regulating smad3/p-smad3 , accompanied by the decrease of Ccr and the increase of col-IV , FN , the content of collagen fibers , RTBM , RTIAW and P10323 . Meantime , the siRNA experiment showed that smad7 can directly regulate a-SMA and P12830 expression . More importantly , miR-21 inhibitor can not only inhibit EMT and fibrosis but also ameliorate renal structure and function . In conclusion , our results demonstrated that miR-21 overexpression can contribute to TGF-β1-induced EMT by inhibiting target smad7 , and that targeting miR-21 may be a better alternative to directly suppress TGF-β1-mediated fibrosis in DN .

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interacts_with DB00864?

multiple_choice

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 . 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 . Attenuation of the progression of adjuvant-induced arthritis by 3-aminobenzamide treatment . Rheumatoid arthritis ( RA ) is a disease that is still insufficiently controlled by current treatments . Poly(ADP-ribose) polymerase ( PARP ) inhibitors ameliorate immune-mediated diseases in several experimental models , including RA , colitis , experimental autoimmune encephalomyelitis and allergy . Together these findings showed that ADP-ribosylating enzymes , in particular P09874 , play a pivotal role in the regulation of immune responses and may represent a noble target for new therapeutic approaches in immune-mediated diseases . The effect of 3-aminobenzamide ( 3-AB ) , an inhibitor of poly(ADP-ribose) synthetase activity , was evaluated in a mouse model of adjuvant-induced arthritis ( AIA ) on pro-inflammatory cytokines , adhesion molecules , inflammatory mediators and chemokine production/expression in serum and knee joint . Histopathological examination was also done on joint section . Our data demonstrates that 3-AB , 10mg/kg , intraperitoneally ( i.p. ) significantly reduces pro-inflammatory cytokine ( Q16552 , P01375 -α and P60568 ) and chemokine ( P13500 and MIP-2 ) production/expression , accompanied by amelioration of the disease as indicated by reduced paw swelling and arthritic scores and was associated with a significant reduction of P19320 and P05362 expression in the knee joint . Moreover , the expression of inflammatory mediators ( P35228 , P35354 , P08253 , P14780 ) and joint histological inflammatory damage was also markedly decreased . The results of this study suggest that P09874 inhibitor may play a role in the inflammatory arthritic process after administration of 3-AB may be a beneficial therapeutic approach . P00747 activation initiated by single-chain urokinase-type plasminogen activator . Potentiation by U937 monocytes . The binding of urokinase-type plasminogen activators ( u-PA ) to receptors on various cell types has been proposed to be an important feature of many cellular processes requiring extracellular proteolysis . We have investigated the effect of single-chain u-PA binding to the monocyte-like cell line U937 on plasminogen activation . A 16-fold acceleration of the activation of plasminogen was observed at optimal concentrations of single-chain u-PA . This potentiation was abolished by the addition of either DB00513 or the amino-terminal fragment of u-PA , thus demonstrating the requirement for specific binding of both single-chain u-PA and plasminogen to the cells . The mechanism of the enhancement of plasmin generation appears to be due primarily to an increase in the rate of feedback activation of single-chain u-PA to the more active two-chain u-PA by cell-bound plasmin , initially generated by single-chain u-PA . This increased activity of the plasminogen activation system in the presence of U937 cells provides a mechanism whereby u-PAs may exert their influence in a variety of cell-associated proteolytic events . Cannabinoid inhibition of adenylate cyclase-mediated signal transduction and interleukin 2 ( P60568 ) expression in the murine T-cell line , EL4. P60568 . Cannabinoid receptors negatively regulate adenylate cyclase through a pertussis toxin-sensitive GTP-binding protein . In the present studies , signaling via the adenylate cyclase/ DB02527 pathway was investigated in the murine thymoma-derived T-cell line , EL4. P60568 . Northern analysis of EL4. P60568 cells identified the presence of 4-kilobase CB2 but not P21554 receptor-subtype mRNA transcripts . Southern analysis of genomic DNA digests for the CB2 receptor demonstrated identical banding patterns for EL4. P60568 cells and mouse-derived DNA , both of which were dissimilar to DNA isolated from rat . Treatment of EL4. P60568 cells with either cannabinol or Delta9-THC disrupted the adenylate cyclase signaling cascade by inhibiting forskolin-stimulated DB02527 accumulation which consequently led to a decrease in protein kinase A activity and the binding of transcription factors to a CRE consensus sequence . Likewise , an inhibition of phorbol 12-myristate 13-acetate ( PMA ) /ionomycin-induced interleukin 2 ( P60568 ) protein secretion , which correlated to decreased P60568 gene transcription , was induced by both cannabinol and Delta9-THC . Further , cannabinoid treatment also decreased PMA/ionomycin-induced nuclear factor binding to the AP-1 proximal site of the P60568 promoter . Conversely , forskolin enhanced PMA/ionomycin-induced AP-1 binding . These findings suggest that inhibition of signal transduction via the adenylate cyclase/ DB02527 pathway induces T-cell dysfunction which leads to a diminution in P60568 gene transcription . Regulation of hyaluronan binding by F-actin and colocalization of P16070 and phosphorylated ezrin/radixin/moesin ( P41161 ) proteins in myeloid cells . Proinflammatory cytokines such as P01375 up-regulate the expression of the cell adhesion molecule , P16070 , and induce hyaluronan ( HA ) binding in peripheral blood monocytes ( PBM ) . Here we show that in PBM , P01375 induced cytoskeletal rearrangement , increased threonine phosphorylation of P41161 proteins , and induced the redistribution and colocalization of phospho- P41161 proteins ( P- P41161 ) with P16070 . In the myeloid progenitor cell line , KG1a , hyaluronan binding occurred in the pseudopod where P16070 , P- P41161 , and F-actin were highly localized . DB08818 binding correlated with high expression of both P16070 and P- P41161 clustered in a single pseudopod . Disruption of polymerized actin reduced hyaluronan binding in both PBM and KG1a cells and abolished P16070 clustering and the pseudopod in KG1a cells . The pseudopod was not required for the clustering of P16070 , the colocalization with P- P41161 , or hyaluronan binding . However , treatment with a kinase inhibitor abolished P41161 phosphorylation and reduced hyaluronan binding . Furthermore , expression of P16070 lacking the putative P41161 binding site resulted in reduced hyaluronan binding . Taken together , these data suggest that P16070 -mediated hyaluronan binding in human myeloid cells is regulated by P- P41161 and the actin cytoskeleton . Role of cardiovascular aldosterone in hypertension . DB04630 plays an important role in the pathogenesis of cardiovascular disease . We have reported that aldosterone is synthesized in cardiovascular tissues and local aldosterone synthesis plays important roles for hypertension and cardiac hypertrophy . High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP . Plasma aldosterone concentrations and P06703 were decreased by high salt intake in SHRSP . DB04630 production , the expression of P19099 mRNA and angiotensin II receptor AT1R mRNA in blood vessels were significantly increased by high salt intake . These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the vascular tissue in SHRSP , which may contribute to the development of malignant hypertension in salt-loaded SHRSP . However , there are several reports of conflicting data . P08235 ( MR ) binding is tightly regulated by the enzyme 11beta-hydroxysteroid dehydrogenase type 2 ( 11beta-HSD2 ) which selectively metabolizes glucocorticoids to inactive metabolites , thus allowing for MR activation by aldosterone . We have reported that decreased 11beta-HSD2 in blood vessels in Dahl salt-sensitive ( DS ) rats , a model for salt-sensitive hypertension . Local aldosterone excess may play a significant role in the salt sensitivity and development of hypertension . High sodium intake decreased circulating rennin-angiotensin-aldosterone system and increased blood pressure and cardiac hypertrophy in DS rats , which were prevented by the treatment with a selective MR antagonist , eplerenone . DB00700 also improved endothelial nitric oxide synthase ( P29474 ) activity and P29474 mRNA expression in blood vessels in DS rats . These results further suggest that not only circulating aldosterone but also local aldosterone is of critical importance in the pathophysiology of cardiovascular disorders . 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 . 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 . FK506-binding protein 12 modulates μ-opioid receptor phosphorylation and protein kinase C(ε)-dependent signaling by its direct interaction with the receptor . Protein kinase C ( PKC ) activation plays an important role in morphine-induced μ-opioid receptor ( P35372 ) desensitization and tolerance development . It was recently shown that receptor phosphorylation by G protein-coupled receptor kinase regulates agonist-dependent selective signaling and that inefficient phosphorylation of P35372 leads to PKCε activation and subsequent responses . Here , we demonstrate that such receptor phosphorylation and PKCε activation can be modulated by FK506-binding protein 12 ( P62942 ) . Using a yeast two-hybrid screen , P62942 was identified as specifically interacting with P35372 at the Pro(353) residue . In human embryonic kidney 293 cells expressing P35372 , the association of P62942 with P35372 decreased the agonist-induced receptor phosphorylation at DB00133 (375) . The morphine-induced PKCε activation and the recruitment of PKCε to the P35372 signaling complex were attenuated both by P62942 short interfering RNA ( siRNA ) treatment and in cells expressing P35372 with a P353A mutation ( OPRM1P353A ) , which leads to diminished activation of PKC-dependent extracellular signal-regulated kinases . Meanwhile , the overexpression of P62942 enabled etorphine to activate PKCε . Further analysis of the receptor complex demonstrated that morphine treatment enhanced the association of P62942 and calcineurin with the receptor . The blockade of the P62942 association with the receptor by the siRNA-mediated knockdown of endogenous P62942 or the mutation of Pro(353) to Ala resulted in a reduction in PKCε and calcineurin recruitment to the receptor signaling complex . The receptor-associated calcineurin modulates P35372 phosphorylation , as demonstrated by the ability of the calcineurin autoinhibitory peptide to increase the receptor phosphorylation . Thus , the association of P62942 with P35372 attenuates the phosphorylation of the receptor and triggers the recruitment and activation of PKCε . 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 . NOS Inhibition Modulates Immune Polarization and Improves Radiation-Induced Tumor Growth Delay . DB00435 synthases ( NOS ) are important mediators of progrowth signaling in tumor cells , as they regulate angiogenesis , immune response , and immune-mediated wound healing . Ionizing radiation ( IR ) is also an immune modulator and inducer of wound response . We hypothesized that radiation therapeutic efficacy could be improved by targeting NOS following tumor irradiation . Herein , we show enhanced radiation-induced ( 10 Gy ) tumor growth delay in a syngeneic model ( C3H ) but not immunosuppressed ( Nu/Nu ) squamous cell carcinoma tumor-bearing mice treated post-IR with the constitutive NOS inhibitor N(G)-nitro-l-arginine methyl ester ( L-NAME ) . These results suggest a requirement of T cells for improved radiation tumor response . In support of this observation , tumor irradiation induced a rapid increase in the immunosuppressive Th2 cytokine P22301 , which was abated by post-IR administration of L-NAME . In vivo suppression of P22301 using an antisense P22301 morpholino also extended the tumor growth delay induced by radiation in a manner similar to L-NAME . Further examination of this mechanism in cultured Jurkat T cells revealed L-NAME suppression of IR-induced P22301 expression , which reaccumulated in the presence of exogenous NO donor . In addition to L-NAME , the guanylyl cyclase inhibitors ODQ and thrombospondin-1 also abated IR-induced P22301 expression in Jurkat T cells and Q14201 -1 macrophages , which further suggests that the immunosuppressive effects involve P29474 . Moreover , cytotoxic Th1 cytokines , including P60568 , IL12p40 , and IFNγ , as well as activated CD8(+) T cells were elevated in tumors receiving post-IR L-NAME . Together , these results suggest that post-IR NOS inhibition improves radiation tumor response via Th1 immune polarization within the tumor microenvironment . Immunization strategies to augment oral vaccination with DNA and viral vectors expressing HIV envelope glycoprotein . Induction of mucosal immunity to the human immunodeficiency virus ( HIV ) envelope ( env ; gp160 ) glycoprotein has been demonstrated with orally administered recombinant vaccinia virus ( rVV ) vectors and poly(DL-lactide-co-glycolide) ( P00747 ) -encapsulated plasmid DNA expressing gp160 . In this study , we investigated the effect of an oral DNA-prime/rVV-boost vaccine regimen in conjunction with adjuvants on the level of gp160-specific cellular and humoral responses in BALB/c mice . We demonstrated that DNA priming followed by a booster with rVV expressing gp160 ( vPE16 ) significantly augmented env-specific immunity in systemic and mucosal tissues of the immunized mice . Association of vPE16 with liposomes and coadministration of liposome-associated beta-glucan lentinan or P60568 /Ig-encoded plasmid DNA-encapsulated in P00747 microparticles triggered the optimal cell-mediated immune ( CMI ) responses . Lentinan was found to increase env-specific type 1 cytokine production and cytotoxic T-lymphocyte ( CTL ) activities but had no effect on humoral responses . On the other hand , P60568 /Ig-mediated increases in both type 1 and 2 activities were associated with higher levels of env-specific CTL and antibody responses . Results of these studies demonstrated the effectiveness of oral vaccines with DNA and rVV vectors in conjunction with immunomodulators in inducing specific immune responses in systemic and mucosal tissues . 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 peptidyl-prolyl isomerase Pin1 regulates granulocyte-macrophage colony-stimulating factor mRNA stability in T lymphocytes . Cytokine production is associated with both the normal and pathologic inflammatory response to injury . Previous studies have shown that the immunosuppressants cyclosporin A or FK506 , which interact with the peptidyl-propyl isomerases cyclophilin A and FK506-binding protein ( P62942 ) , respectively , block cytokine expression . A third member of the peptidyl-propyl isomerase family , Pin1 is expressed by immune and other cells . Pin1 has been implicated in cell cycle progression , is overexpressed in human tumors , and may rescue neurons from tau-associated degeneration . However , the role of Pin1 in the immune system remains largely unknown . In this study , we analyze the role of Pin1 in GM- P04141 expression by human PBMC and P01730 + lymphocytes . We show that Pin1 isomerase activity is necessary for activation-dependent , GM- P04141 mRNA stabilization , accumulation , and protein secretion , but not non-AU-rich elements containing cytokine mRNAs , including TGF-beta and P05112 . Mechanistically , Pin1 mediated the association of the AU-rich element-binding protein , Q14103 , with GM- P04141 mRNA , which determined the rate of decay by the exosome . 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 ) . Role of nicotinamide in DNA damage , mutagenesis , and DNA repair . DB02701 is a water-soluble amide form of niacin ( nicotinic acid or vitamin B3 ) . Both niacin and nicotinamide are widely available in plant and animal foods , and niacin can also be endogenously synthesized in the liver from dietary tryptophan . DB02701 is also commercially available in vitamin supplements and in a range of cosmetic , hair , and skin preparations . DB02701 is the primary precursor of nicotinamide adenine dinucleotide ( NAD(+) ) , an essential coenzyme in DB00171 production and the sole substrate of the nuclear enzyme poly-ADP-ribose polymerase-1 ( P09874 ) . Numerous in vitro and in vivo studies have clearly shown that P09874 and NAD(+) status influence cellular responses to genotoxicity which can lead to mutagenesis and cancer formation . This paper will examine the role of nicotinamide in the protection from carcinogenesis , DNA repair , and maintenance of genomic stability . 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 . 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 .

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interacts_with DB05039?

multiple_choice

Neu1 sialidase and matrix metalloproteinase-9 cross-talk regulates nucleic acid-induced endosomal TOLL-like receptor-7 and -9 activation , cellular signaling and pro-inflammatory responses . The precise mechanism(s) by which intracellular TOLL-like receptors ( TLRs ) become activated by their ligands remains unclear . Here , we report a molecular organizational G-protein coupled receptor ( GPCR ) signaling platform to potentiate a novel mammalian neuraminidase-1 ( Neu1 ) and matrix metalloproteinase-9 ( P14780 ) cross-talk in alliance with neuromedin B GPCR , all of which form a tripartite complex with TLR-7 and -9 . siRNA silencing Neu1 , P14780 and neuromedin-B GPCR in RAW-blue macrophage cells significantly reduced Q9NYK1 imiquimod- and Q9NR96 ODN1826-induced NF-κB ( NF-κB-pSer(536) ) activity . Tamiflu , specific P14780 inhibitor , neuromedin B receptor specific antagonist BIM23127 , and the selective inhibitor of whole heterotrimeric G-protein complex O43521 -46174 significantly block nucleic acid-induced TLR-7 and -9 MyD88 recruitment , NF-κB activation and proinflammatory TNFα and P13500 cytokine responses . For the first time , Neu1 clearly plays a central role in mediating nucleic acid-induced intracellular TLR activation , and the interactions involving P28336 - P14780 -Neu1 cross-talk constitute a novel intracellular TLR signaling platform that is essential for NF-κB activation and pro-inflammatory responses . 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 . Autocrine P13501 signaling promotes invasion and migration of CD133+ ovarian cancer stem-like cells via NF-κB-mediated P14780 upregulation . The concept of cancer stem cells ( CSCs ) proposes that solely CSCs are capable of generating tumor metastases . However , how CSCs maintain their invasion and migration abilities , the most important properties of metastatic cells , still remains elusive . Here we used CD133 to mark a specific population from human ovarian cancer cell line and ovarian cancer tissue and determined its hyperactivity in migration and invasion . Therefore , we labeled this population as cancer stem-like cells ( CSLCs ) . In comparison to CD133- non-CSLCs , chemokine P13501 and its receptors , P32246 , P51677 , and P51681 , were consistently upregulated in CSLCs , and most importantly , blocking of P13501 , P32246 , or P51677 effectively inhibits the invasive capacity of CSLCs . Mechanistically , we identified that this enhanced invasiveness is mediated through nuclear factor κB ( NF-κB ) activation and the consequently elevated P14780 secretion . Our results suggested that the autocrine activation of P32246 and P51677 by P13501 represents one of major mechanisms underlying the metastatic property of ovarian CSLCs . 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 . 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 . 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 . Endo- P07237 is required for TNFα-induced angiogenesis . Protein disulfide isomerase ( P07237 ) and its homologs are oxidoreductases facilitating protein folding in the ER . Endo- P07237 ( also termed Q8NBS9 ) is highly expressed in endothelial cells . It belongs to the P07237 family but its physiological function is largely unknown . We studied the role of Endo- P07237 in endothelial angiogenic responses . Stimulation of human umbilical vein endothelial cells ( with TNFα ( 10ng/ml ) increased P27361 /2 phosphorylation . This effect was largely attenuated by Endo- P07237 siRNA , whereas JNK and p38 Q96HU1 kinase phosphorylation was Endo- P07237 independent . Similarly , TNFα-stimulated NF-κB signaling determined by IκBα degradation as well as TNFα-induced ICAM expression was unaffected by Endo- P07237 siRNA . The action of Endo- P07237 was not mediated by extracellular thiol exchange or cell surface P07237 as demonstrated by nonpermeative inhibitors and P07237 -neutralizing antibody . Moreover , exogenously added P07237 failed to restore P27361 /2 activation after Endo- P07237 knockdown . This suggests that Endo- P07237 acts intracellularly potentially by maintaining the Ras/Raf/MEK/ P29323 pathway . Indeed , knockdown of Endo- P07237 attenuated Ras activation measured by G-LISA and Raf phosphorylation . P29323 activation influences gene expression by the transcriptional factor AP-1 , which controls P14780 and cathepsin B , two proteases required for angiogenesis . TNFα-stimulated P14780 and cathepsin B induction was reduced by silencing of Endo- P07237 . Accordingly , inhibition of cathepsin B or Endo- P07237 siRNA blocked the TNFα-stimulated angiogenic response in the spheroid outgrowth assays . Moreover ex vivo tube formation and in vivo Matrigel angiogenesis in response to TNFα were attenuated by Endo- P07237 siRNA . In conclusion , our study establishes Endo- P07237 as a novel , important mediator of AP-1-driven gene expression and endothelial angiogenic function . DB03615 inhibits the chaperone activity of protein disulfide isomerase . In the process of screening of proteins binding to ribostamycin in bovine liver using the affinity column chromatography , we found that ribostamycin inhibited the chaperone activity of protein disulfide isomerase ( P07237 ) , but it did not inhibit the isomerase activity . P07237 was identified by SDS-PAGE , Western blotting , and N-terminal amino acid sequence analysis . A 100:1 molar ratio of ribostamycin to P07237 was almost sufficient to completely inhibit the chaperone activity of P07237 . The binding affinity of ribostamycin to purified bovine P07237 was determined by the Biacore system , which gave a K(D) value of 3.19 x 10(-4) M . This suggests that ribostamycin binds to region distinct from the CGHC motif of P07237 . This is the first report to describe the inhibitor of the chaperone activity of P07237 . 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 . The cannabinoid DB00470 mediates inhibition of macrophage chemotaxis to RANTES/ P13501 : linkage to the CB2 receptor . The chemotactic response of murine peritoneal macrophages to RANTES/ P13501 was inhibited significantly following pretreatment with DB00470 ( THC ) , the major psychoactive component in marijuana . Significant inhibition of this chemokine directed migratory response was obtained also when the full cannabinoid agonist CP55940 was used . The CB2 receptor-selective ligand O-2137 exerted a robust inhibition of chemotaxis while the P21554 receptor-selective ligand ACEA had a minimal effect . The THC-mediated inhibition was reversed by the CB2 receptor-specific antagonist SR144528 but not by the P21554 receptor-specific antagonist SR141716A . In addition , THC treatment had a minimal effect on the chemotactic response of peritoneal macrophages from CB2 knockout mice . Collectively , these results suggest that cannabinoids act through the CB2 receptor to transdeactivate migratory responsiveness to RANTES/ P13501 . Furthermore , the results suggest that the CB2 receptor may be a constituent element of a network of G protein-coupled receptor signal transductional systems , inclusive of chemokine receptors , that act coordinately to modulate macrophage migration . 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 ) DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . 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 . DB00087 in the treatment of chronic lymphocytic leukemia . DB00087 is a humanized therapeutic monoclonal antibody ( MAb ) that recognizes the P31358 antigen , expressed on normal and neoplastic lymphocytes , monocytes , and natural killer cells . In 2001 , alemtuzumab was approved in the US and Europe to treat B-cell chronic lymphocytic leukemia ( CLL ) that had been treated previously with alkylating agents and was refractory to fludarabine . In heavily pretreated patients this MAb is able to produce response rates of about 40 % , and in symptomatic , previously untreated patients response rates of more than 80 % can be achieved . DB00087 can also be used in patients with CLL as a preparative regimen for stem cell transplantation ( P09683 ) and to prevent graft versus host disease . Moreover its in vivo use before or after P09683 may also potentially result in depletion of residual leukemia cells , especially in the autologous setting . Adverse events associated with alemtuzumab include acute first-dose reaction , hematologic toxicity , and infectious complications . Usually they are predictable , manageable , and acceptable in the context of CLL . However , in a significant percentage of patients , cytomegalovirus reactivation occurs during alemtuzumab therapy , and routine weekly monitoring with the polymerase chain reaction methodology is indicated . Moreover , antiviral and antibacterial prophylaxis is mandatory . [ The role of matrix metalloproteinases and their inhibitors in pathogenesis of pancreatic pseudocysts ] . The investigation was conducted in 47 patients , operated on for pancreatic pseudocysts ( PP ) . Activity of matrix metalloproteinases ( P14780 ) and content of their tissue inhibitor ( P16035 ) were determined in the blood serum for estimation of inflammatory factors , hypoxia severity and state of the pancreatic tissue reconstruction . High activity of P14780 and P16035 in presence of PP types I and II was noted in patients , what , probably , is caused by compensation reaction , directed towards inhibition of the collagen system destruction ( predominantly of collagen type IV ) and prevention of further reconstruction of pancreatic connective tissue . While progressing of pancreatic fibrosis the P14780 activity and the P16035 level have lowered in comparison with these indices while its absence . In PP type III the P14780 activity was by 83.6 % higher , than in a control group , but , by 51.4 and 35.1 % lower , than in PP types I and IV . In all the patients endothelial dysfunction with endothelial injury was observed , witnessed by significant rising of the P15692 content in the blood serum . It have created favorable conditions for pancreatic tissue remodeling while parenchymal defect have been constituted by tissue , owing lower level of organization , including a cicatricial one . In cases of cellular repeated affection more activation of pancreatic stellate cells and enhancement of production of extracellular matrix component were noted . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . An absolute role of the PKC-dependent NF-kappaB activation for induction of P14780 in hepatocellular carcinoma cells . Matrix metalloproteinases ( MMPs ) play an important role in inflammation , tumor cell invasion , and metastasis . We found that phorbol-12-myristate-13-acetate ( PMA ) -stimulated invasion of the hepatocellular carcinoma ( HCC ) SNU-387 and SNU-398 cells and that PMA induced the secretion of P14780 in the cells , but did not induce the secretion of P08253 . The PMA-induced P14780 secretion was abolished by treatment of a pan-protein kinase C ( PKC ) inhibitor , GF109203X , and an inhibitor of NF-kappaB activation , sulfasalazine , and partly inhibited by treatment of inhibitors of P29323 pathway , PD98059 and U0126 . In addition , the PMA-stimulated activation of the P14780 promoter was completely inhibited by a mutation of the NF-kappaB site within the P14780 promoter , but not completely by mutations of two AP-1 sites . Moreover , the P14780 induction by P14210 and P01375 was also completely inhibited by GF109203X and sulfasalazine , but not by PD98059 and U0126 . These data demonstrate that the PKC-dependent NF-kappaB activation is absolute for P14780 induction and that the PKC-dependent P29323 activation devotes to increase the expression level of P14780 , in HCC cells . [ Spiral computed tomography with functional tests in diagnosis of chronic obstructive diseases of the lungs ] . The potential of spiral computed tomography ( P09683 ) and high resolution computed tomography ( HRST ) in diagnosis of chronic obstructive pulmonary diseases ( P48444 ) is described . Semiotic sings of bronchial asthma ( BA ) and chronic obstructive bronchitis ( P00156 ) are specified . Informative value of P09683 in differential diagnosis of BA and P00156 including early stage of the diseases is analysed . An updated technique of P09683 and HRCT of the lungs in P48444 patients is presented . 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 . Sinusoidal endothelial cells as an early target for hepatic toxicants . Recent studies demonstrate that the hepatic sinusoidal endothelial cells ( SEC ) are a sensitive direct target for early toxicity to acetaminophen ( paracetamol , DB00316 ) and this toxicity is exacerbated following a single and multiple week-end type alcoholic binge(s) . SEC become swollen and begin to lose the ability to endocytose FITC- Q2LD37 , a ligand for the scavenger receptor , as early as 30 minutes after the administration of DB00316 . Gaps through the SEC appear to be formed by the destruction and/or coalescence of fenestrae and are seen as early as 2 hrs after the administration of DB00316 which is prior to any evidence of injury to parenchymal cells . The gaps permit red blood cells to penetrate into the Space of Disse . Subsequently , the sinusoid may collapse or disintegrate reducing blood flow . The gaps are larger and more frequent in ethanol binged animals subsequently treated with DB00316 . Similar gaps are seen in the early stages of hepatic venoocclusive disease . Administration of a NO donor or a P08253 and P14780 inhibitor minimizes endothelial injury and red blood cell penetration into the Space of Disse . The injury is exacerbated when an inhibitor of P29474 is administered and minimized when P35228 is inhibited suggesting a protective role for constitutive NO derived from SEC . Both NO and MMPs are known to affect the cytoskeleton of SEC which in turn affects the formation and maintenance of the fenestrae .

MH_train_1428

MH_train_1428

MH_train_1428

interacts_with DB04839?

multiple_choice

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 . 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 . Randomised clinical trial : herbal extract DB05767 in active ulcerative colitis - a double-blind comparison with sustained release mesalazine . BACKGROUND : Andrographis paniculata is an herbal mixture used to treat inflammatory diseases . An extract of the herb , DB05767 , inhibits P01375 -α and IL-1β , and prevents colitis in animal models . AIM : To determine the efficacy and safety of DB05767 in patients with mild-to-moderate ulcerative colitis . METHODS : A randomised , double-blind , multicentre , 8-week parallel group study was conducted using DB05767 1200 mg/day compared with 4500 mg/day of slow release mesalazine ( mesalamine ) granules in patients with mild-to-moderately active ulcerative colitis . Disease activity was assessed at baseline and every 2 weeks for clinical response , and at baseline and 8 weeks by colonoscopy . RESULTS : One hundred and twenty patients at five centres in China were randomised and dosed . Clinical remission and response were seen in 21 % and 76 % of DB05767 -treated patients , and 16 % and 82 % of mesalazine-treated patients . By colonoscopy , remission and response were seen in 28 % and 74 % of DB05767 -treated patients and 24 % and 71 % of mesalazine-treated patients , respectively . There was no significant difference between the two treatment groups . CONCLUSION : DB05767 may be an efficacious alternative to mesalazine in ulcerative colitis . 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 . 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 . 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 . 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 . Receptor kinase inhibitors target NSCLC : two antibodies and a small-molecule MET inhibitor . Joining cetuximab , sorafenib , afatinib , intedanib , and crizotinib in phase III development for non-small cell lung cancer ( NSCLC ) are ramucirumab ( developed by ImClone , a subsidiary of Lilly ) , necitumumab ( developed by ImClone and Bristol-Myers Squibb ) , and tivantinib ( ARQ 197 , developed by ArQule and Daiichi Sankyo ) . DB09559 is a second-generation anti- P00533 monoclonal antibody ( mAb ) similar to cetuximab . Enrollment has been stopped in one of two necitumumab phase III trials because of safety concerns . DB05578 is an anti- P35968 mAb targeting the same pathway as bevacizumab . Although the phase II safety data for ramucirumab appear better than the data for necitumumab , fewer phase III data are available . Tivantinib is a highly selective , orally available MET tyrosine kinase inhibitor . MET is overexpressed in 61 % of NSCLC cases . Although tivantinib is the last of the three agents discussed here to enter phase III , its phase II results are the most robust . P10275 -induced tumor suppressor , B2CW77 , inhibits breast cancer growth and transcriptionally activates p53/p73-mediated apoptosis in breast carcinomas . P10275 ( AR ) expression by immunohistochemistry correlates with better prognosis and survival among breast cancer patients . We and others have shown that AR inhibits proliferation and induces apoptosis in breast cancer cells . However , the mechanism of AR 's anti-tumor effect in breast cancer is still not fully understood . Our recent study indicates that AR upregulates expression of tumor suppressor gene P60484 by promoter activation in breast cancer . B2CW77 , encoding B2CW77 protein , is a newly identified gene , which shares a bidirectional promoter with P60484 and is transcribed in the opposite direction . So far , the function of B2CW77 has never been studied in tumorigenesis . Here , we define B2CW77 as a tumor suppressor in breast carcinomas , which inhibits tumor growth and invasiveness . After analyzing 188 normal breast and 1247 malignant breast cancer tissues , we observed the loss of B2CW77 in multiple breast cancer subtypes and this decreased B2CW77 expression associates with tumor progression and increasing histological grade in invasive carcinomas . We characterize B2CW77 , for the first time , as a transcription factor , directly promoting the expression of P04637 and O15350 , with consequent elevated apoptosis and cell cycle arrest in breast cancer cells . We demonstrate , in vitro and in murine xenograph models , that both B2CW77 and P60484 are AR-target genes , mediating androgen-induced growth inhibition and apoptosis in breast cancer cells . Our observations suggest that B2CW77 might be used as a potential prognostic marker and novel therapy target for breast carcinomas . 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 . P10275 expression induces P09038 , FGF-binding protein production , and P09038 release in prostate carcinoma cells : role of P09038 in growth , survival , and androgen receptor down-modulation . BACKGROUND : Alterations in fibroblast growth factors ( FGFs ) production and/or FGF receptors expression have been described to play key roles in prostate tumor progression , particularly in androgen-independent tumors . However , the role of androgen receptor ( AR ) in altering FGF-mediated growth and survival of prostatic neoplastic cells has not been completely defined . In this study , we investigated the alterations in P09038 production and utilization by the PC3 cell line , after transfection with a full-length AR . METHODS : P05230 ,2,7 , FGF-binding protein ( Q14512 ) production and FGF receptor ( FGFR ) 1-4 expression were investigated by polymerase chain reaction ( PCR ) and Western blot analysis . RESULTS : De novo AR expression by PC3 cells restores P21802 IIIb isoform expression and sensitivity to P21781 and P09038 . Androgen stimulation induces AR+ PC3 clones to secrete Q14512 , likely responsible for activation and mobilization from the extracellular matrix of the high amounts of P09038 produced by the same cells . In addition to the effects on cell proliferation , P09038 maintains the survival of AR+ PC3 clones through a positive modulation of the Bcl-2 protein and down-modulates AR protein expression , allowing the escape of selected clones from androgen regulation . CONCLUSION : In the presence of an active AR , the combined production of P09038 and Q14512 may play an important role in the progression of prostate cancer through the selection of AR- clones expressing high levels of Bcl-2 . Circulating hormones and hypothalamic energy balance : regulatory gene expression in the Lou/C and Wistar rats . To ascertain the mechanisms underlying low caloric intake and low body weight in the Lou/C rat , the circulating hormone levels and gene expression of hypothalamic peptides and receptors important in energy balance and the induction of suppressor of cytokine signalling 3 ( O14543 ) gene expression in response to leptin challenge were compared with Wistar rats . Plasma leptin levels were lower in the Lou/C rat , as were levels of rat corticosterone , DB00024 and DB00451 but not DB00279 . Ghrelin levels were higher in the Lou/C rat . Total leptin receptor ( Ob-R ) and the long form of the leptin receptor ( Ob- P06400 ) gene expression were lower in the arcuate ( ARC ) and ventromedial nuclei ( VMN ) in Lou/C rat . Q92847 expression in the ARC and VMN was lower in Lou/C than in Wistar rats . However , agouti gene-related peptide ( AgRP ) and neuropeptide Y ( P01303 ) gene expression were higher in the Lou/C rat . There was no difference in the level of cocaine- and amphetamine-regulated transcript gene expression in the ARC , but both were higher in the paraventricular nuclei of the Lou/C breed . There was no difference in Ob-R gene expression in , or [(125)I]leptin binding to , the choroid plexus . O14543 mRNA induction in response to leptin was lower in the Lou/C rat . This study reveals that the comparatively low plasma leptin , DB00024 and DB00451 levels , and high ghrelin levels together with high levels of AgRP and P01303 gene expression seen in the Lou/C rat are indicative of a strong drive to eat and decreased energy expenditure , which are in direct opposition to the comparatively low body weight and adiposity of this rat strain . Signaling and antiproliferative effects of type I and II gonadotropin-releasing hormone receptors in breast cancer cells . DB00644 receptors ( DB00644 -Rs ) mediate direct antiproliferative effects on hormone-dependent cancer cells . DB00644 -Rs can be grouped according to ligand specificity ( for DB00644 and -II ) , and there is evidence that type II DB00644 ligands and/or receptors can inhibit proliferation . Type I DB00644 -Rs ( e.g. human and sheep ) lack the C-terminal tails found in other G protein-coupled receptors including type II DB00644 -Rs ( e.g. Xenopus ; XGnRH-R ) . This underlies the remarkable resistance of type I DB00644 -Rs to desensitization and may be important for chronic effects on proliferation . To test this , we have compared the antiproliferative effects of DB00644 -Rs expressed in MCF7 breast cancer cells using recombinant adenovirus ( Ad ) . Endogenous DB00644 -Rs were not detected , but infection with Ad-expressing sheep DB00644 -Rs ( sGnRH-R ) facilitated proliferation inhibition by DB06719 , and maximum inhibition required only 10,000-20,000 sGnRH-Rs . XGnRH-Rs were much less efficient at inhibiting proliferation and were internalized faster than sGnRH-Rs . Thus , the type II P30968 is less efficient at inhibiting proliferation , presumably because it is rapidly desensitized and/or internalized . Moreover , comparisons of human P30968 , sGnRH-R , and XGnRH-R , as well as chimeric receptors ( type I DB00644 -Rs with C-terminal tails from XGnRH-Rs ) , revealed that C-terminal tail addition increases receptor expression and thereby increases the efficiency with which the vector facilitates the antiproliferative effect . 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 . Identification of novel genetic alterations in samples of malignant glioma patients . Glioblastoma is the most frequent and malignant human brain tumor . High level of genomic instability detected in glioma cells implies that numerous genetic alterations accumulate during glioma pathogenesis . We investigated alterations in AP-PCR DNA profiles of 30 glioma patients , and detected specific changes in 11 genes not previously associated with this disease : Q86UP9 , Q13326 , Q13639 , P05556 , P31327 , P07225 , P55259 , Q9UJ96 , Q08499 , Q8N743 , and Q14642 . Further correlations revealed that 8 genes might play important role in pathogenesis of glial tumors , while changes in P55259 , Q9UJ96 and Q8N743 should be considered as passenger mutations , consequence of high level of genomic instability . Identified genes have a significant role in signal transduction or cell adhesion , which are important processes for cancer development and progression . According to our results , Q86UP9 might be characteristic of primary glioblastoma , Q13326 , Q13639 , P05556 , P31327 , P07225 and Q14642 were detected predominantly in anaplastic astrocytoma , suggesting their role in progression of secondary glioblastoma , while alterations of Q08499 seem to have important role in development of both glioblastoma subtypes . Some of the identified genes showed significant association with p53 , p16 , and P00533 , but there was no significant correlation between loss of P60484 and any of identified genes . In conclusion our study revealed genetic alterations that were not previously associated with glioma pathogenesis and could be potentially used as molecular markers of different glioblastoma subtypes . 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 . P10275 expression in breast cancer patients tested for P38398 and P51587 mutations . AIM : To assess the expression of receptors for androgen ( AR ) , oestrogen ( ER ) and progesterone ( PR ) as well as human epidermal growth factor receptor type 2 ( Her-2/neu ) status of breast carcinomas in breast cancer susceptibility gene ( BRCA ) P38398 /2 mutation carriers and P38398 /2 negative tested women . METHODS : One hundred and thirty-five breast cancers in women tested for P38398 /2 mutations . Screening for P38398 and P51587 mutations was performed by direct sequencing of all P38398 and P51587 exons as well as the surrounding intronic sequences . Additionally , BRCA genes were analysed with multiplex ligation-dependent probe amplification . Consecutive paraffin sections were examined immunohistochemically for AR , ER , PR and Her-2/neu . RESULTS : Of the 135 tumours , 43 ( 32 % ) were P38398 -related , 18 ( 13 % ) were P51587 -related and 74 ( 55 % ) were P38398 /2-negative . Seventy-two per cent of the P38398 -related , 22 % of the P51587 -related and 12 % of the P38398 /2-negative tumours were triple ( ER , PR , Her2neu ) -negative . Eighty-four per cent of P38398 mutated cancers were high-grade ( P46379 ) tumours . ARs were expressed in 30 % ( 13 of 43 ) of P38398 -related , in 78 % ( 14 of 18 ) in P51587 -related tumours and in 76 % ( 56 of 74 ) in P38398 /2 negative tumours . Twenty-one per cent of ER-negative P38398 -related tumours expressed androgen receptors . CONCLUSION : Approximately one in five P38398 mutated breast cancers negative for ER and PR express androgen receptors . Modulation of AR might open a new avenue for treating these high-risk cancers . 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 . 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 . 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 . Inhibition of angiogenesis and tumor growth by murine DB00054 , the parent antibody of c7E3 Fab ( abciximab ; ReoPro ) . Angiogenesis plays an essential role in the growth and dissemination of solid tumor cancers . The expression of endothelial cell integrin alpha(v)beta3 has been shown to increase during vascular proliferation associated with human tumors . Selective antagonists of alpha(v)beta3 can block angiogenesis and tumor growth by inducing programmed cell death in proliferating endothelial cells . Monoclonal antibody DB00054 , an antagonist of the human , but not murine , integrins alpha(v)beta3 and alphaIIbbeta3 ( P08514 /IIIa ) , inhibits platelet aggregation . It is the parent antibody of a mouse/human chimeric antibody fragment approved for adjunctive therapy of patients undergoing percutaneous coronary interventions to prevent ischemic complications ( c7E3Fab ; abciximab ; ReoPro ) . To evaluate the potential of 7E3 to inhibit human angiogenesis and tumor growth independent of its antiplatelet effects , we established integrin alpha(v)beta3-negative human melanoma tumors in full-thickness human skin grafted onto SCID mice . The resulting tumors induce a human angiogenic response as assessed by the immunoreactivity of vascular cells with monoclonal antibodies specific for human CD31 . Administration of 7E3 prevented or significantly inhibited the growth of tumors , and this effect correlated with a significant reduction in the number of blood vessels supplying the tumors . These results support the previous findings that blockade of integrin alpha(v)beta3 inhibits angiogenesis and tumor growth and indicates that dual inhibitors of alpha(v)beta3 and alphaIIbbeta3 are effective in blocking tumor growth and angiogenesis . Radiosensitization by inhibition of IkappaB-alpha phosphorylation in human glioma cells . To assess the role of nuclear factor kappaB ( NFKB ) in cellular radiosensitivity , three different IkappaB-alpha ( also known as P25963 ) expression plasmids , i.e. , S-IkappaB ( mutations at ( 32 , 36 ) DB00133 ) , Y-IkappaB ( a mutation at (42) DB00135 ) , and SY-IkappaB , were constructed and introduced into human brain tumor M054 cells . The clones were named as M054-S8 , M054- P28062 and M054-SY4 , respectively . Compared to the parental cell line , M054-S8 and M054- P28062 cells were more sensitive to X rays while M054-SY4 cells exhibited the greatest sensitivity . After treatment with N-acetyl- DB00149 - DB00149 -norleucinal , a proteasome inhibitor , the X-ray sensitivity of M054-S8 and M054-SY4 cells did not change , while that of M054- P28062 cells and the parental cells was enhanced . An increase in X-ray sensitivity accompanied by a decrease in translocation of NFKB to the nucleus in parental cells was observed after treatment with pervanadate , an inhibitor of tyrosine phosphatase , as well as in M054-S8 and M054-SY4 cells . Repair of potentially lethal damage ( PLD ) was observed in the parental cells but not in the clones . Four hours after irradiation ( 8 Gy ) , the expression of P04637 and phospho-p53 ( (15) DB00133 ) was induced in the parental cells but not in M054-S8 , M054- P28062 or M054-SY4 cells . Our data suggest that inhibition of IkappaB-alpha phosphorylation at serine or tyrosine acts independently in sensitizing cells to X rays . NFKB may play a role in determining radiosensitivity and PLD repair in malignant glioma cells ; P04637 may also be involved . Forced expression of P38936 in P08514 - P38936 transgenic mice induces abnormalities in the proliferation of erythroid and megakaryocyte progenitors and primitive hematopoietic cells . OBJECTIVE : P38936 ( P38936 /Cip/kip) and p27(Kip1) are cyclin-dependant kinase inhibitors controlling cell-cycle exit and differentiation of numerous cell types . Among hematopoietic cells , megakaryocytes express high levels of P38936 , while in erythroid cells , p27(Kip1) is predominant . As P38936 and p27 could display overlapping functions and as megakaryocytes and erythroid cells derive from a bipotent progenitor , we developed an in vivo model to determine the specific role of P38936 in controlling the proliferation/differentiation balance of erythroid and megakaryocytic progenitors . METHODS : Transgenic mice that overexpressed P38936 under the control of the human P08514 promoter in early progenitors and along megakaryocytic differentiation were generated . Different subsets of hematopoietic progenitors ( BFU and CFU ) and primitive cells ( CAFC , LTC-IC ) were analyzed by methylcellulose assay . Phenotypic evolution and clonogenic properties of the lin(-) population were analyzed along erythroid and megakaryocytic differentiation . RESULTS : We observed P38936 ectopic expression in early hematopoietic progenitors ( lin(-)Sca(+) ) , megakaryocytes , and , to a lesser extent , erythroid cells . This expression induced an important decrease in the number of CFU-MK , BFU-E , CFU-E , primitive progenitors ( CAFC day 35 ) , and LTC-IC , but did not affect the maturation process of these cells and the blood cell count . CONCLUSIONS : We show that variation of P38936 expression level changes the fate of hematopoietic cells by favoring either proliferation or differentiation pathways . This effect of P38936 is exerted not only at the level of primitive progenitors but also in more mature progenitors . However , in vivo , a systemic compensation mechanism is most likely activated in response to variations of the flow of progenitor production .

MH_train_1429

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interacts_with DB04868?

multiple_choice

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 . [ Detection of a high-affinity prostaglandin I2 binding site in the human thyroid ] . This study is concerned with the identification and the pharmacological properties of P43119 binding sites on human thyroid membrane fractions . Scatchard analysis is not linear , revealing a high- and a low-affinity receptor binding site . ( 3 H ) DB01088 binding experiments were performed under various clinical conditions : in thyroid cancer the low-affinity binding sites disappear totally and the specific high-affinity binding sites are diminished according to the grade of differentiation of the cancer . An alteration in Bmax and Kd is also observed in cold nodules , in Hashimoto 's and Riedl 's thyroiditis and in hyperthyroidism , whereas hot nodules exhibit an increase in both the receptor subpopulations . The data provide evidence for specific DB01240 binding sites and support the suggestion of a direct regulatory key-role of DB01240 in thyroid intermediary metabolism . P43119 induces P42224 and P40763 phosphorylations in human erythroleukemia cells : a mechanism requiring PTX-insensitive G proteins , P29323 and JNK . The ability of the human prostacyclin receptor ( hIP ) to regulate the activities of signal transducers and activators of transcription ( STATs ) has not yet been documented . In the present study , we have delineated the mechanism by which hIP induces P40763 phosphorylations in human erythroleukemia ( HEL ) cells . Stimulation of endogenous hIP by its specific agonist , cicaprost , resulted in P40763 Tyr705 and Ser727 phosphorylations in a time- and concentration-dependent manner . Cicaprost-induced P40763 Tyr705 and Ser727 phosphorylations were resistant to pertussis toxin ( PTX ) treatment , suggesting that these responses were mediated through PTX-insensitive G proteins . In addition , extracellular signal-regulated kinase ( P29323 ) and c-Jun N-terminal kinase ( JNK ) , but not p38 MAPK , were shown to be phosphorylated by cicaprost in a time- and concentration-dependent manner via PTX-insensitive G proteins . The levels of the interaction between P40763 , P29323 and JNK were enhanced by cicaprost treatment . The involvement of P04049 , Q02750 /2 and JNK in cicaprost-induced phosphorylations of P40763 was illustrated by the use of their selective inhibitors . In contrast , p38 MAPK did not appear to be required . Similar observations were obtained with P42224 upon stimulation by cicaprost . Taken together , these results demonstrate for the first time that hIP activation by cicaprost can lead to P42224 and P40763 phosphorylations via signaling pathways involving PTX-insensitive G proteins , P29323 and JNK . Regulation of phosphodiesterase-4 ( DB05876 ) expression in mouse brain by repeated antidepressant treatment : comparison with rolipram . Cyclic nucleotide phosphodiesterase-4 ( DB05876 ) is a component of signaling pathways involved in the mediation of antidepressant activity . Of the four DB05876 subtypes , Q08499 appears to be of particular importance , given the finding that Q08499 -deficient mice exhibit an antidepressant-like behavioral phenotype . In mouse hippocampus and cerebral cortex , the effects of repeated treatment with the antidepressants desipramine and fluoxetine or the DB05876 inhibitor rolipram on the expression of Q08499 was compared to that of P27815 and Q07343 , the other two subtypes expressed in the brain . Expression of Q08499 was increased by all drugs tested , with the exception of desipramine in hippocampus . By contrast , these treatments affected P27815 and Q07343 expression differentially . In hippocampus , antidepressants increased P27815 and decreased Q07343 , whereas ROL decreased P27815 and did not change Q07343 . In cerebral cortex , antidepressants increased P27815 and did not change Q07343 , whereas ROL did not change P27815 and increased Q07343 . 3H- DB01954 binding was increased in cytosolic , but not in membrane , fractions of cerebral cortex by all drugs tested ; there were no changes observed in hippocampus . Overall , the present results suggest some species-dependence of the regulation of DB05876 subtypes , based on data obtained previously using rats . They also suggest that the Q08499 subtype may be of particular importance as an antidepressant target in that it is regulated by repeated treatment with both norepinephrine and serotonin reuptake inhibitors as well as by the DB05876 inhibitor rolipram , drugs that produce antidepressant effects via different neuropharmacological mechanisms . The P11274 / P00519 -inhibitors imatinib , nilotinib and dasatinib differentially affect NK cell reactivity . In chronic myeloid leukemia ( CML ) , P11274 / P00519 -mediated oncogenic signaling can be targeted with the P11274 / P00519 -inhibitors Imatinib , DB04868 and Dasatinib . However , these agents may also affect anti-tumor immunity . Here , we analyzed the effects of the 3 P11274 / P00519 -inhibitors on natural killer ( NK ) cell reactivity . Exposure of CML cells ( K562 , Meg-01 ) to pharmacological concentrations of Imatinib , DB04868 and Dasatinib diminished expression of ligands for the activating immunoreceptor P26718 to a similar extent . This resulted in comparably reduced NK cell cytotoxicity and P01579 production . When direct effects on NK cell responses to K562 and primary CML cells as well as activating cytokines were studied , Dasatinib was found to abrogate NK cytotoxicity and cytokine production . DB04868 did not alter cytotoxicity but , at high levels , impaired NK cytokine production , while Imatinib had no direct influence on NK cell reactivity . Of note , DB04868 , but not the other P11274 / P00519 -inhibitors increased cell death within the preferentially cytokine-secreting CD56(bright)CD16(-) NK cell subset , which may , at least in part , serve to explain the effect of DB04868 on NK cytokine production . Analysis of NK cell signaling revealed that Dasatinib inhibited proximal signaling events leading to decreased phosphorylation of PI3K and P29323 that are crucial for NK cell reactivity . Imatinib and DB04868 , in contrast , showed no relevant effect on NK cell PI3K or P29323 activity . In light of the potential role of NK cells in the immunesurveillance of residual leukemia and for future combinatory immunotherapeutic approaches , our data indicate that choice and dosing of the most suitable P11274 / P00519 -inhibitor for a given patient require careful consideration . Saengmaeksan inhibits inflammatory mediators by suppressing O43353 /caspase-1 activation . BACKGROUND AND OBJECTIVES : Saengmaeksan ( P52788 ) is a Korean herbal prescription consisting of three different herbal drugs : Liriopis Tuber ( tuber of Liriope platyphylla , Liliaceae ) , DB01404 Radix ( root of Panax ginseng ) and Schisandrae Fructus ( fruit of Schisandra chinensis ) . P52788 is commonly used in Korea to treat various diseases that involve the respiratory and cardiovascular systems . However , to date , the mechanism underlying the anti-inflammatory effects of P52788 is not clearly understood . In this study , we attempt to determine the effects of P52788 on lipopolysaccharide ( LPS ) -induced inflammatory responses in mouse peritoneal macrophages . METHODS : Cell viability was measured by using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay , and nitric oxide ( NO ) levels were measured by using Griess reagent . The tumor necrosis factor ( P01375 ) -α and interleukin ( IL ) -6 levels secreted by the cells were measured using a modified enzyme-linked immunosorbent assay . Expression of cyclooxygenase ( P36551 ) -2 and nuclear factor-kappa B ( NF-κB ) , respectively was investigated using a western blot analysis . A caspase colorimetric assay kit was used to assay enzymatic caspase-1 activity . RESULTS : The findings of this study showed that P52788 reduced P01375 -α and P05231 production induced by LPS . During the inflammatory process , P35354 and NO levels were increased in mouse peritoneal macrophages , but P52788 decreased the enhanced levels of P35354 and the production of NO . In addition , P52788 suppressed the activation of NF-κB and receptor interacting protein-2/caspase-1 . DISCUSSION AND CONCLUSION : Our results provide novel insights into the pharmacological actions of P52788 , a molecule that can potentially be exploited in the treatment of inflammatory diseases . 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 . 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 . 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 . Genome wide mapping reveals Q07343 as an P60568 induced P42229 target gene in activated human PBMCs and lymphoid cancer cells . P60568 is the primary growth factor for promoting survival and proliferation of activated T cells that occurs following engagement of the Janus Kinase (JAK)1-3/and Signal Transducer and Activator of Transcription ( P35610 ) 5 signaling pathway . P42229 has two isoforms : P42229 and P51692 ( commonly referred to as P42229 ) which , in T cells , play redundant roles transcribing cell cycle and survival genes . As such , inhibition of P42229 by a variety of mechanisms can rapidly induce apoptosis in certain lymphoid tumor cells , suggesting that it and its target genes represent therapeutic targets to control certain lymphoid diseases . To search for these molecules we aligned P60568 regulated genes detected by Affymetrix gene expression microarrays with the P42229 cistrome identified by chip-on-ChIP analysis in an P60568 -dependent human leukemia cell line , Kit225 . Select overlapping genes were then validated using qRT(2)PCR medium-throughput arrays in human PHA-activated PBMCs . Of 19 putative genes , one key regulator of T cell receptor signaling , Q07343 , was identified as a novel target , which was readily up-regulated at the protein level ( 3 h ) in P60568 stimulated , activated human PBMCs . Surprisingly , only purified CD8+ primary T-cells expressed Q07343 , but not P01730 + cells . Moreover , Q07343 was found to be highly expressed in P01730 + lymphoid cancer cells , which suggests that it may represent a physiological role unique to the CD8+ and lymphoid cancer cells and thus might represent a target for pharmaceutical intervention for certain lymphoid diseases . 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 . 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 . DB08860 prevents DB01221 -induced retinal ganglion cell death by suppressing leukocyte recruitment . Excitotoxicity is a major cause of retinal ganglion cell ( RGC ) death during ischemic diseases such as vessel occlusion and diabetic retinopathy . However , the underlying mechanisms are not well understood . Statins , inhibitors of the P04035 , have neuroprotective effects in addition to their original role in lowering cholesterol . We hypothesize that pitavastatin , a recently introduced potent statin , is protective against N-methyl-d-aspartic acid ( DB01221 ) -induced RGC death . DB08860 , administered by gavage , abolished DB01221 -induced loss of RGCs . To elucidate the mechanisms underlying the neuroprotective effect of pitavastatin , we investigated its impact on inflammation . DB01221 increased the expression of interleukin-1beta and P01375 , and endothelial adhesion molecules , including P05362 , and induced leukocyte accumulation in the retinal vessels . DB08860 significantly reduced DB01221 -induced leukocyte accumulation and up-regulation of endothelial adhesion molecules , whereas cytokine expression was unaffected . Systemic blockade of P05362 in wild-type mice or absence of P05107 in gene-deficient ( P05107 (-/-) ) mice significantly suppressed DB01221 -induced leukocyte accumulation and RGC death . These findings suggest a novel and causative role for inflammatory leukocyte recruitment in DB01221 -induced excitotoxicity . Furthermore , we show the novel neuroprotective effect of statins against excitotoxicity-induced RGC death . Statins or other anti-inflammatory agents may thus have therapeutic benefits in excitotoxicity-associated neuronal diseases through blockade of leukocyte recruitment . 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 . 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 . SV40 infection associated with rituximab treatment after kidney transplantation in nonhuman primates . BACKGROUND : A regimen consisting of polyclonal anti-T-cell antibody , sirolimus ( Q86TD4 ) , and donor bone marrow ( DBM ) infusion induces robust transplantation tolerance to skin allografts in mice . We investigated the effect of a similar regimen in a nonhuman primate ( NHP ) model . METHODS : Cynomolgus macaques ( Macaca fascicularis ) were transplanted with mismatched kidney allografts . Recipients were treated with 7 doses of antithymocyte globulin ( DB00098 , day 1 to 9 ) , sirolimus , and DBM infusion ( day 14 ) . Anti- P11836 antibody , rituximab , was given on days 0 and 5 . RESULTS : A regimen of DB00098 , 30 days of Q86TD4 , and DBM infusion induced significantly greater prolongation of graft survival with a mean survival time of 88 days compared with the control regimen ( no DBM ) with an mean survival time of 53 days ( P=0.022 ) . Unlike the murine skin allograft model , all grafts were rejected within 111 days . A combination of DB00098 , continuous Q86TD4 , and rituximab caused graft and systemic SV40 infection and failed to achieve further extension of graft survival . C4d deposition was observed in 50 % of recipients as early as 18 days , suggesting antidonor antibody production . A transient , low-to-moderate degrees of multilineage chimerism was observed after DBM infusion . Treatment with DB00098 resulted in profound depletion of P01730 + and CD8+ T cells , whereas addition of rituximab achieved prolonged ( up to 3 months ) depletion of P11836 + B cells . CONCLUSION : The DB00098 , Q86TD4 , and DBM protocol is simple and produces long-term kidney allograft survival in NHP although additional treatment modalities may be necessary for induction of long-term tolerance . Distinct interaction of nilotinib and imatinib with P-Glycoprotein in intracellular accumulation and cytotoxicity in CML Cell Line K562 cells . DB04868 , a second-generation tyrosine kinase inhibitor ( TKI ) , has been approved for first-line chronic myeloid leukemia ( CML ) treatment . The improved clinical response of nilotinib over that of the first generation TKI , imatinib , has been thought to be a result of its high potency of inhibition of P11274 - P00519 kinase . This study aimed to characterize differences between nilotinib and imatinib in the intracellular accumulation and cytotoxic effect on the CML cell line K562 . Accumulation of nilotinib in K562 cells was from 4.7- to 9.0-fold higher than that of imatinib . The cytotoxic effect of nilotinib on K562 cells was 14.2-fold higher than that of imatinib . Inhibition experiments in K562 cells , and examination of the cellular uptake using influx transporter-transfected human embryonic kidney ( P29320 ) 293 cells , suggested that the influx transporters OCT1 and P46721 , which have been reported to mediate accumulation of imatinib in CML cells , contributed little to the uptake of nilotinib . DB04868 was found to accumulate in imatinib-resistant K562 ( K562/IM ) cells overexpressing the efflux transporter P-glycoprotein ( P-gp ) , although cytotoxic assays showed that K562/IM cells displayed 20000-fold greater resistance to nilotinib over the parent K562 cells . In conclusion , the present findings suggest that intracellular accumulation of nilotinib in CML cells contributes to its clinical response and efficacy in CML patients . Although nilotinib has been reported to be effective against imatinib-resistant P00519 kinase mutants , the drug could not overcome imatinib resistance acquired by P-gp-overexpression . These results imply that classification of mechanisms of drug resistance is important for suitable strategies to treat imatinib-resistant CML patients . The stop transfer sequence of the human UDP-glucuronosyltransferase 1A determines localization to the endoplasmic reticulum by both static retention and retrieval mechanisms . Human UDP-glucuronosyltransferase 1A ( P22309 ) isoforms are endoplasmic reticulum ( ER ) -resident type I membrane proteins responsible for the detoxification of a broad range of toxic phenolic compounds . These proteins contain a C-terminal stop transfer sequence with a transmembrane domain ( TMD ) , which anchors the protein into the membrane , followed by a short cytosolic tail ( CT ) . Here , we investigated the mechanism of ER residency of P22309 mediated by the stop transfer sequence by analysing the subcellular localization and sensitivity to endoglycosidases of chimeric proteins formed by fusion of P22309 stop transfer sequence ( TMD/CT ) with the ectodomain of the plasma membrane P01730 reporter protein . We showed that the stop transfer sequence , when attached to C-terminus of the P01730 ectodomain was able to prevent it from being transported to the cell surface . The protein was retained in the ER indicating that this sequence functions as an ER localization signal . Furthermore , we demonstrated that ER localization conferred by the stop transfer sequence was mediated in part by the KSKTH retrieval signal located on the CT . Interestingly , our data indicated that P22309 TMD alone was sufficient to retain the protein in ER without recycling from Golgi compartment , and brought evidence that organelle localization conferred by P22309 TMD was determined by the length of its hydrophobic core . We conclude that both retrieval mechanism and static retention mediated by the stop transfer sequence contribute to ER residency of P22309 proteins . Induction of P22309 and P20813 by an antimitogenic factor in HepG2 cells is mediated through suppression of cyclin-dependent kinase 2 activity : cell cycle-dependent expression . P14210 ( P14210 ) , an antimitogenic factor for HepG2 cells , increased mRNA and protein levels of P22309 and P20813 , as well as the endogenous cyclin-dependent kinase ( CDK ) inhibitors p16 , P38936 , and p27 in HepG2 cells but not in HuH6 , Caco2 , or MCF7 cells . Treatment with 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene ( U0126 ) ( an extracellular signal-regulated kinase inhibitor ) suppressed the P14210 -induced expression of P22309 and P20813 , as well as p16 , P38936 , and p27 in HepG2 cells . The CDK inhibitor roscovitine also enhanced the expression of P22309 , P20813 , and P08684 . Transfection of anti- P24941 siRNA led to elevated levels of P22309 , P20813 , and P08684 in HepG2 and SW480 cells , whereas anti- P11802 small interfering RNA ( siRNA ) did not significantly enhance the expression of these enzymes . In fact , P24941 activity was decreased in P14210 -treated HepG2 cells . In cells arrested in S phase by a thymidine block and then released into a synchronous cell cycle , there was a clear dissociation among the activation of P24941 and the expression of P22309 , P20813 , and P08684 . Furthermore , the induction of P08684 but not P22309 or P20813 mRNA expression by roscovitine was repressed in pregnane X receptor ( O75469 ) siRNA-transfected HepG2 cells . Transfection with constitutive androstane receptor siRNA or O75469 siRNA in HepG2 cells did not repress the P14210 -stimulated expression of P22309 mRNA . Taken together , our results show that the expression of P22309 and P20813 is negatively regulated through a P24941 signaling pathway linked to cell cycle progression in HepG2 and SW480 cells , the mechanism of which may differ from that of P08684 expression through O75469 phosphorylated by P24941 .

MH_train_1430

MH_train_1430

MH_train_1430

interacts_with DB08815?

multiple_choice

A double-blind , placebo-controlled outpatient trial of pergolide for cocaine dependence . Results of preclinical studies suggest that pergolide , a mixed D(1)/ P14416 agonist , may be useful in treating cocaine dependence . To empirically investigate this possibility , we conducted a 5-year , double-blind , placebo-controlled clinical trial of two doses of pergolide ( 0.05 and 0.25 mg bid ) in subjects with cocaine dependence and combined cocaine/alcohol dependence . Data analysis was performed on an intent to treat population ( N=358 ) and a per protocol population ( N=108 ) with urine drug screens ( UDS ) used as the main outcome measure . There were no significant effects on UDS at either pergolide dose . DB01186 had no significant effect on alcohol use in the comorbid alcohol/cocaine dependence group . DB01186 does not appear to have clinical value in the treatment of cocaine dependence or in decreasing alcohol use in cocaine-dependent individuals at the presently studied doses . P60568 -dependent augmentation of the anti-TNP antibody production by sodium butyrate in cultured murine splenic B cells . Previously we found that sodium butyrate ( NaBu ) markedly enhanced production of the antibody specific for a T-cell-dependent antigen , sheep red blood cells ( SRBC ) in murine splenocytes ( Kishiro , Y. , Ueda , K. , Fujiwara , M. and Yamamoto , I. , Jpn J. Phamacol. , 1994 66 , 369-376 . To gain a better understanding of the target cells for NaBu 's action on antibody responses , we have utilized the T-cell-independent antigen , trinitrophenyl-lypopolysaccharide ( TNP-LPS ) as a stimulant and have examined an effect of NaBu on the anti-TNP antibody production in vitro . NaBu markedly increased the anti-TNP plaque-forming cell ( P27918 ) responses in murine whole splenocytes , but not in murine splenic B cells . Addition of T-cells or the concanavalin A supernatant ( CAS ) from murine splenocytes to the B cell cultures completely restored the enhancing effect of NaBu . This effect of CAS was totally blocked by an anti-interleukin ( IL ) -2 antibody and partially by an anti- P01584 or anti- P05112 antibody . The full enhancing effect of NaBu was also detected when P60568 was added to the B cell cultures , while P60568 alone had no stimulatory effect on the control P27918 response . P01584 alone significantly stimulated the antibody production and adding NaBu to this P01584 -supplemented culture caused a further increase . Neither P05112 alone nor NaBu plus P05112 had any effect on the P27918 response . NaBu did not affect the expression of the P60568 receptor alpha- and beta-chains in B cells stimulated with TNP-LPS . These results suggest that NaBu is an agent that promotes B cell differentiation in vitro in an P60568 -dependent manner . 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 . 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 . The dynamic mobility of histone H1 is regulated by cyclin/CDK phosphorylation . The linker histone H1 is involved in maintaining higher-order chromatin structures and displays dynamic nuclear mobility , which may be regulated by posttranslational modifications . To analyze the effect of H1 tail phosphorylation on the modulation of the histone 's nuclear dynamics , we generated a mutant histone H1 , referred to as M1-5 , in which the five cyclin-dependent kinase phosphorylation consensus sites were mutated from serine or threonine residues into alanines . P12004 E/ P24941 or cyclin A/ P24941 can not phosphorylate the mutant in vitro . Using the technique of fluorescence recovery after photobleaching , we observed that the mobility of a green fluorescent protein ( GFP ) -M1-5 fusion protein is decreased compared to that of a GFP-wild-type H1 fusion protein . In addition , recovery of H1 correlated with P24941 activity , as GFP-H1 mobility was decreased in cells with low P24941 activity . Blocking the activity of P24941 by P38936 expression decreased the mobility of GFP-H1 but not that of GFP-M1-5 . Finally , the level and rate of recovery of cyan fluorescent protein ( P27918 ) -M1-5 were lower than those of P27918 -H1 specifically in heterochromatic regions . These data suggest that P24941 phosphorylates histone H1 in vivo , resulting in a more open chromatin structure by destabilizing H1-chromatin interactions . [ Treatment of neovascular age-related macular degeneration with antiangiogenic drugs ] . Age-related macular degeneration ( ARMD ) remains a leading cause of blindness in the western world . Several clinical forms of the disease are recognized , whereas choroidal neovascularization ( CNV ) represents an important manifestation suitable for treatment . The treatment of CNV has been a major focus of research in the past decades , and the first evidence-based established therapy was laser photocoagulation , which reduces the risk of visual loss in extrafoveal lesions . In the late 90 's photodynamic therapy has been established as an efficient method for the treatment of predominantly classic and occult CNV . Additional therapies such as macular translocation , submacular surgery , and indocyanine-mediated prothrombosis are currently under investigation in large-scale clinical trials . Molecular biology has recently provided a better comprehension of the pathogenesis of ARMD , and vascular endothelial growth factor ( P15692 ) was recognized as key mediator in the angiogenesis of CNV-formation . Therefore , the pharmacological approach rose as a key research area to treat CNV . The first FDA-approved agent for CNV-therapy is aptamer pegaptanib sodium ( Macugen ) , which inactivates the key angiogenic isoform VEGF165 . Additional P15692 -blockers such as ranibizumab RhuFab V2 ( Lucentis ) and bevacizumab ( DB00112 ) are under evaluation in major clinical studies . Impressive results of intravitreal bevacizumab were released recently . Moreover , the steroid-derived DB05288 as well as the corticosteroid triamcinolone acetate have been proposed as methods for treatment of wet-ARMD . This paper presents the rationale and principles of the pharmacologic antiangiogenic therapy for CNV in ARMD . 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 . 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 . [ Effects of various growth factors on the growth of trophoblast cells in long-term culture ] . Trophoblasts taken from placental tissue of the 1st trimester and molar tissue , and P30793 -1 ( gestational choriocarcinoma cell line ) cells were cultured in collagen coated dishes . The medium used was a mixture of DME and Ham 's F-12 ( 1:1 ) , containing P01133 , PDGF , insulin , GM- P04141 , IL-1 , -2 , -3 , PGE1 and DB00917 in various concentrations . 3H-TdR uptake of the cultured cells was measured as a marker of cell growth . The growth of normal trophoblasts was enhanced by PDGF or insulin , and remarkably by P01133 + PDGF + insulin + GM- P04141 . The growth of molar trophoblasts was accelerated by P01133 or insulin . Under these culture conditions , normal trophoblasts have been successfully cultured and maintained for 12 months and molar trophoblasts for 9 months to date . The cultured cells were identified with trophoblasts by immunohistochemical staining with P62158 5.2 monoclonal antibody . No effect of growth factors was observed in P30793 -1 cells . 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 . 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 . 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 . Reinvestigation of carrier transport properties in liquid crystalline 2-phenylbenzothiazole derivatives . We have reinvestigated the charge carrier transport properties in a liquid crystal of 2-(4'-heptyloxyphenyl)-6-dodecylthiobenzothiazole ( 7O- P10721 - P28222 ) , for which the electronic conduction was first established in rodlike liquid crystals and for which the highest hole mobility in the smectic A ( SmA ) phase ever achieved was reported . We found that 7O- P10721 - P28222 exhibited three crystal phases , one of which appeared in a limited temperature range of 10 degrees just below the phase transition temperature from the SmA phase . In this crystal phase , nondispersive transient photohole currents were observed in time-of-flight experiments , and its hole mobility was determined to be 8 x 10(-3) cm(2)/Vs , slightly higher than that reported previously in the SmA phase . For the SmA phase , however , the hole mobility was 1 x 10(-4) cm(2)/Vs . Furthermore , we established the electron transport in the SmA phase of purified 7O- P10721 - P28222 , whose mobility was the same as the hole mobility in that phase . In order to confirm generality of the new findings in 7O- P10721 - P28222 , we investigated the carrier transport properties of its derivative having a short hydrocarbon chain , 2-(4'-heptyloxyphenyl)-6-butylthiobenzothiazole ( 7O- P10721 -S4 ) , and obtained comparable results . The present results correct a mistake in the previous report and give an idea of what a typical mobility in the SmA phase is . On the basis of these results , we discuss what determines the charge carrier mobility in smectic mesophases . Preclinical evaluation of nilotinib efficacy in an imatinib-resistant P10721 -driven tumor model . The novel P10721 inhibitor nilotinib is currently being evaluated for its clinical utility in the treatment of gastrointestinal stromal tumor . However , the effects of nilotinib in cells expressing commonly occurring P10721 mutations remain to be fully defined . The aim of this study was therefore to investigate the efficacy of nilotinib against cells expressing imatinib-sensitive or imatinib-resistant P10721 mutations and to evaluate [ (18)F ] DB09150 -positron emission tomography ( DB09150 -PET ) imaging as a biomarker of nilotinib response in vivo . DB04868 inhibited the proliferation of imatinib-responsive V560G- P10721 P18627 -P1 and imatinib-resistant D816V- P10721 P18627 -P1 cells with a GI(50) of 4.9 and 630 nmol/L , respectively , whereas apoptosis studies revealed that nilotinib and imatinib were equipotent against the V560G cell line . In contrast , although 10 micromol/L nilotinib induced > 50 % apoptosis in the D816V cells at 16 hours , 10 micromol/L imatinib had no effect on cell survival at 24 hours . Syngeneic DBA2/J mice bearing P18627 -P1- P10721 tumors were evaluated for response to nilotinib by DB09150 -PET . V560G- P10721 P18627 -P1 tumor DB09150 uptake was significantly reduced compared with baseline levels following 2 days of nilotinib treatment . In contrast , no effect of nilotinib was observed on tumor growth or DB09150 -PET uptake into D816V tumors despite intratumoral drug levels reaching in excess of 10 micromol/L at 4 hours after dosing . Biomarker analysis revealed the inhibition of P10721 phosphorylation in V560G but not D816V tumors . These findings show the in vivo activity of nilotinib in the treatment of tumors bearing V560G- P10721 but not D816V- P10721 and the utility of DB09150 -PET imaging to assess tumor response to this agent . 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 . 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 . Stimulation of cloned human serotonin P28221 beta receptor sites in stably transfected P13671 glial cells promotes cell growth . The involvement of serotonin P28221 beta receptor sites was investigated in the growth of rat P13671 glial cells permanently transfected with a gene encoding a human P28221 beta receptor . The 5-HT receptor identity of control and transfected P13671 glial/ P28221 beta cells was determined by reverse transcription-polymerase chain reaction using primers specific for rat P08908 , rat P28222 , rat P28221 alpha , human P28221 beta , and rat 5- Q13049 receptor genes . Constitutive mRNA for 5- Q13049 receptors was present in control and transfected P13671 glial/ P28221 beta cells , whereas mRNA for P28221 beta receptor sites was only present in the transfected P13671 glial/ P28221 beta cell line . 5-HT inhibited forskolin-stimulated cyclic AMP formation and promoted cell growth , in contrast to the absence of any measurable effect in pcDNA3 plasmid-transfected and nontransfected P13671 glial cells . The 5-HT effects could be mimicked by sumatriptan ( EC50 = 44-76 nM ) and were totally and partially blocked by methiothepin ( IC50 = 9 nM ) and GR 127,935 ( 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol-3-yl)-biphenyl-4-carbox yli c acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide ; IC50 = 97 pM ) , respectively . No effect on cell growth was measured with the 5-HT2 receptor agonist DOI [ 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ; 10 microM ] , suggesting that 5- Q13049 receptors are not involved in the 5-HT-stimulated P13671 glial/ P28221 beta cell growth . Dibutyryl-cyclic AMP ( 0.3 mM ) -treated cultures did not show sumatriptan-promoted cell growth , indicating an inhibitory role for cyclic AMP in the cell growth mediated by P28221 beta receptor sites. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . 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 ) Effects of lurasidone on executive function in common marmosets . Cognitive impairment is one of the major symptoms of schizophrenia , and is considered largely due to dysfunctions in the prefrontal cortex ( P27918 ) . DB08815 , a novel atypical antipsychotic agent with high binding affinity for dopamine D2 , serotonin P34969 , 5- Q13049 and P08908 receptors has been reported to have superior efficacy in rodents ' models of cognitive impairment . However , the beneficial effect of lurasidone on cognitive impairment has not been evaluated in non-human primates . In this study , we investigated the effect of lurasidone on executive function , which is one of the cognitive domains , in common marmosets and compared the results to those of other antipsychotics . The effects of lurasidone , haloperidol , olanzapine , risperidone , quetiapine and clozapine on executive function were evaluated in naïve marmosets using the object retrieval with detours ( ORD ) task . Before drug treatment , marmosets ' success rates in the easy trial of the test were almost 90 % . However , maximum success in the difficult trial of the task reached only 50 % after 8 days of training . DB00502 , olanzapine and risperidone decreased correct performance even in the easy trial of the task . All drugs , except lurasidone , impaired success rate in the difficult trial . On the other hand , lurasidone dose-dependently increased marmosets ' success rates in the difficult trial with significant effect at 10mg/kg . In conclusion , we have shown in this study that lurasidone , unlike conventional antipsychotics , improves cognition associated with executive function in common marmosets . These findings suggest that lurasidone would be more useful for treatment of schizophrenia cognitive impairment than other antipsychotics . 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 . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents .

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DB05595 in lung cancer . DB00158 is essential for proliferating cells and folate transport pathways and folate-dependent metabolic processes show promise as targets for anti-neoplastic therapy . DB00158 receptor α ( P15328 ) , a folate transporter , is an attractive target for anti-neoplastic therapy due to its high affinity for folate , restricted range of expression in normal tissue and differential over-expression in malignant tissue . P15328 is expressed in non-small cell lung cancer , with a higher expression in adenocarcinoma compared with squamous cell carcinoma . DB05595 is a monoclonal antibody targeting P15328 which in pre-clinical studies led to cytotoxicity of P15328 -expressing cells , inhibited tumor growth in animal models and showed limited reactivity with normal tissue . In phase I/II trials , farletuzumab was well tolerated as a single-agent and in combination , without additive toxicity with chemotherapy . An ongoing phase II , double blind , placebo-controlled study is evaluating farletuzumab in patients with P15328 expressing metastatic adenocarcinoma of lung . 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 . DB06366 ( Perjecta ) for P04626 -positive metastatic breast cancer . Epidermal growth factor as a local mediator of the neurotrophic action of vitamin B(12) ( cobalamin ) in the rat central nervous system . We have recently demonstrated that the myelinolytic lesions in the spinal cord ( SC ) of rats made deficient in vitamin B(12) ( cobalamin ) ( Cbl ) through total gastrectomy ( TG ) are tumor necrosis factor-alpha ( P01375 ) -mediated . We investigate whether or not permanent Cbl deficiency , induced in the rat either through TG or by chronic feeding of a Cbl-deficient diet , might modify the levels of three physiological neurotrophic factors-epidermal growth factor ( P01133 ) , vasoactive intestinal peptide ( P01282 ) , and somatostatin ( SS ) -in the cerebrospinal fluid ( P04141 ) of these rats . We also investigated the ability of the central nervous system ( CNS ) in these Cbl-deficient rats to synthesize P01133 mRNA and of the SC to take up labeled Cbl in vivo . Cbl-deficient rats , however the vitamin deficiency is induced , show a selective decrease in P01133 P04141 levels and an absence of P01133 mRNA in neurons and glia in various CNS areas . In contrast , radiolabeled Cbl is almost exclusively taken up by the SC white matter , but to a much higher degree in totally gastrectomized ( O43548 ) rats . Chronic administration of Cbl to O43548 rats restores to normal both the P01133 P04141 level and P01133 mRNA expression in the various CNS areas examined . This in vivo study presents the first evidence that the neurotrophic action of Cbl in the CNS of O43548 rats is mediated by stimulation of the P01133 synthesis in the CNS itself . It thus appears that Cbl inversely regulates the expression of P01133 and P01375 genes in the CNS of O43548 rats . Tissue inhibitors of matrix metalloproteinases are elevated in cerebrospinal fluid of neurodegenerative diseases . Matrix metalloproteinases ( MMPs ) are implicated in the pathogenesis of diseases such as Alzheimer 's Disease ( AD ) and amyotrophic lateral sclerosis ( P35858 ) . Increased expression of P14780 and TIMPs has been reported in postmortem AD and P35858 brain tissue , as well as in P35858 cerebrospinal fluid ( P04141 ) and plasma . Although individual studies of MMP and P01033 expression in P04141 have included AD and P35858 samples , there are no studies comparing the expression of these proteins between neurodegenerative diseases . We measured the levels of matrix metalloproteinases ( MMPs ) -2 and -9 and the tissue inhibitor of MMPs ( e.g. P01033 and P16035 ) in P04141 samples from patients with Parkinson 's Disease ( PD ) , Huntington 's Disease ( HD ) , AD and P35858 as compared to age-matched control patients . There was constitutive expression of the proform of gelatinase A ( proMMP-2 ) on zymography gels in all P04141 samples . Unexpectedly , there was an additional gelatinolytic band at 130 kDa of unknown etiology in the P04141 samples of patients with PD ( 61 % of patients studied ) , AD ( 61 % ) , HD ( 25 % ) and P35858 ( 39 % ) . Levels of P01033 were significantly elevated in P04141 samples from all disease groups . P16035 was significantly increased in P04141 of AD and HD patients . P08253 levels did not differ significantly between groups . These findings show that TIMPs are elevated in the P04141 of patients with neurodegenerative diseases suggesting a potential role of these endogenous inhibitors of matrix metalloproteinases in neurodegenerative diseases . 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 . The urokinase receptor is overexpressed in the AIDS dementia complex and other neurological manifestations . The urokinase-type plasminogen activator ( uPA ) and its receptor ( Q03405 ) play an important role in extracellular matrix degradation and cell migration in the central nervous system ( CNS ) . To investigate the role of the uPA/ Q03405 system in the pathophysiology of acquired immunodeficiency syndrome dementia complex ( ADC ) , we measured soluble Q03405 ( suPAR ) levels in cerebrospinal fluid ( P04141 ) and plasma from human immunodeficiency virus ( HIV ) -1-infected patients and controls . P04141 suPAR levels were significantly higher in HIV-1-infected patients than in controls and in patients with ADC or opportunistic CNS infections ( CNS-OIs ) than in neurologically asymptomatic patients , irrespective of HIV-1 disease stage . The highest levels of suPAR were found in patients with ADC , and among those with CNS-OIs in patients with cytomegalovirus encephalitis or cryptococcosis . Plasma suPAR levels were higher in HIV-1-infected patients than in controls and increased with HIV-1 disease stage regardless of the presence of CNS disease . In patients with ADC or CNS-OIs , P04141 suPAR levels correlated with P04141 HIV-1 RNA , but not with plasma suPAR concentrations . Highly active antiretroviral therapy was associated with a significant and parallel decrease of both P04141 suPAR and HIV-1 RNA . In brain tissue from patients with HIV-1 encephalitis , Q03405 was highly expressed by microglial and multinucleated giant cells staining positively for HIV-1 . The overexpression of Q03405 in the CNS of patients with ADC suggests that the uPA/ Q03405 system may contribute to the tissue injury and neuronal damage in this disease . Carcinomatous meningitis : Leptomeningeal metastases in solid tumors . Leptomeningeal metastasis ( LM ) results from metastatic spread of cancer to the leptomeninges , giving rise to central nervous system dysfunction . Breast cancer , lung cancer , and melanoma are the most frequent causes of LM among solid tumors in adults . An early diagnosis of LM , before fixed neurologic deficits are manifest , permits earlier and potentially more effective treatment , thus leading to a better quality of life in patients so affected . Apart from a clinical suspicion of LM , diagnosis is dependent upon demonstration of cancer in cerebrospinal fluid ( P04141 ) or radiographic manifestations as revealed by neuraxis imaging . Potentially of use , though not commonly employed , today are use of biomarkers and protein profiling in the P04141 . Symptomatic treatment is directed at pain including headache , nausea , and vomiting , whereas more specific LM-directed therapies include intra- P04141 chemotherapy , systemic chemotherapy , and site-specific radiotherapy . A special emphasis in the review discusses novel agents including targeted therapies , that may be promising in the future management of LM . These new therapies include anti-epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors erlotinib and gefitinib in nonsmall cell lung cancer , anti- P04626 monoclonal antibody trastuzumab in breast cancer , anti- P16410 ipilimumab and anti- P15056 tyrosine kinase inhibitors such as vermurafenib in melanoma , and the antivascular endothelial growth factor monoclonal antibody bevacizumab are currently under investigation in patients with LM . Challenges of managing patients with LM are manifold and include determining the appropriate patients for treatment as well as the optimal route of administration of intra- P04141 drug therapy . 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 . Multiple sclerosis during adalimumab treatment in a case with ankylosing spondylitis . Ankylosing spondylitis is a chronic and progressive inflammatory disease involving the sacroiliac joints with HLA- Q8TCY5 positivity in 85 % of the patients and radiologically evidence of sacroiliitis . It is associated with several extraarticular manifestations , but neurological complications are rare . Occurrence of multiple sclerosis in patients with ankylosing spondylitis has been reported in limited cases . DB00051 , a P01375 -α antagonist , offers a significant improvement in ankylosing spondylitis and is considered to be less immunogenic and more tolerable than other P01375 -α blockers . A case of multiple sclerosis coexisted with HLA- Q8TCY5 positive ankylosing spondylitis after treated with adalimumab was reported . Cell-specific regulation of acetylcholinesterase expression under inflammatory conditions . DB03128 ( ACh ) has been shown to exert an anti-inflammatory function by down-modulating the expression of pro-inflammatory cytokines . Its availability can be regulated at different levels , namely at its synthesis and degradation steps . Accordingly , the expression of acetylcholinesterase ( P22303 ) , the enzyme responsible for ACh hydrolysis , has been observed to be modulated in inflammation . To further address the mechanisms underlying this effect , we aimed here at characterizing P22303 expression in distinct cellular types pivotal to the inflammatory response . This study was performed in the human acute leukaemia monocytyc cell line , THP-1 , in human monocyte-derived primary macrophages and in human umbilical cord vein endothelial cells ( HUVEC ) . In order to subject these cells to inflammatory conditions , THP-1 and macrophage were treated with lipopolysaccharide ( LPS ) from E.coli and HUVEC were stimulated with the tumour necrosis factor α ( P01375 -α ) . Our results showed that although P22303 expression was generally up-regulated at the mRNA level under inflammatory conditions , distinct P22303 protein expression profiles were surprisingly observed among the distinct cellular types studied . Altogether , these results argue for the existence of cell specific mechanisms that regulate the expression of acetylcholinesterase in inflammation . Translational PK-PD modelling of molecular target modulation for the AMPA receptor positive allosteric modulator Org 26576 . INTRODUCTION : The α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ( AMPA ) receptor potentiator Org 26576 represents an interesting pharmacological tool to evaluate the utility of glutamatergic enhancement towards the treatment of psychiatric disorders . In this study , a rat-human translational pharmacokinetic-pharmacodynamic ( PK-PD ) model of AMPA receptor modulation was used to predict human target engagement and inform dose selection in efficacy clinical trials . METHODS : Modelling and simulation was applied to rat plasma and cerebrospinal fluid ( P04141 ) pharmacokinetic and pharmacodynamic measurements to identify a target concentration ( EC(80) ) for AMPA receptor modulation . Human plasma pharmacokinetics was determined from 33 healthy volunteers and eight major depressive disorder patients . From four out of these eight patients , P04141 PK was also determined . Simulations of human P04141 levels were performed for several doses of Org 26576 . RESULTS : Org 26576 ( 0.1-10 mg/kg , i.v. ) potentiated rat hippocampal AMPA receptor responses in an exposure-dependant manner . The rat plasma and P04141 PK data were fitted by one-compartment model each . The rat P04141 PK-PD model yielded an EC(80) value of 593 ng/ml ( 90 % confidence interval 406.8 , 1,264.1 ) . The human plasma and P04141 PK data were simultaneously well described by a two-compartment model . Simulations showed that in humans at 100 mg QD , P04141 levels of Org 26576 would exceed the EC(80) target concentration for about 2 h and that 400 mg P55957 would engage AMPA receptors for 24 h . CONCLUSION : The modelling approach provided useful insight on the likely human dose-molecular target engagement relationship for Org 26576 . Based on the current analysis , 100 and 400 mg P55957 would be suitable to provide ' phasic ' and ' continuous ' AMPA receptor engagement , respectively . 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 . 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 . 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 . Use of trifluoroperazine isolates a [(3)H] DB08954 binding site in rat brain membranes with the pharmacology of the voltage-independent ifenprodil site on N-methyl-D-aspartate receptors containing Q13224 subunits . The use of trifluoroperazine in a well washed rat brain membrane preparation revealed [(3)H]ifenprodil binding to a single high affinity state with the pharmacology of N-methyl-D-aspartate ( DB01221 ) receptors containing Q13224 subunits . Inhibition of [(3)H]ifenprodil binding in the presence of trifluoroperazine by 10 NR1a/ Q13224 selective agents was highly correlated with their inhibition at rat NR1a/ Q13224 receptors expressed in Xenopus ooctyes and [(3)H] DB01520 binding to rat brain Q13224 subunit containing DB01221 receptors but not with their inhibition of [(3)H]DTG binding . Allosteric interactions with polyamines , Mg(2+) , Zn(2+) , glutamate , glycine , and their antagonists were consistent with DB01221 receptors with Q13224 subtype pharmacology . The rank order of polyamine inhibition was spermine > spermidine > 1,5-(diethylamino)piperidine > arcaine > agmatine > putrescine . Both spermidine and MgCl(2) shifted the inhibition curve of ifenprodil to the right in a parallel manner , but Mg(2+) did not appear to be additive to spermidine . Glutamate increased and glycine decreased the binding . Conversely , CPP decreased the binding , and MDL 105,519 increased the binding in an agonist reversible manner . The increase with MDL 105,519 and glutamate appeared to be additive as did the decrease with glycine and CPP . Changes in the buffer pH between 6.5 and 8.0 did not affect the affinity of Q13224 agents . DB09202 but not clonidine inhibited the binding . MK-801 and agents from various other pharmacological classes did not significantly inhibit [(3)H]ifenprodil binding . [(3)H] DB08954 binding in the presence of trifluoroperazine appears to be selective for the voltage-independent ifenprodil site on DB01221 receptors containing the Q13224 subunit . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . The 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 . NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC .

MH_train_1432

MH_train_1432

MH_train_1432

interacts_with DB09068?

multiple_choice

Small molecule-driven mitophagy-mediated Q96P20 inflammasome inhibition is responsible for the prevention of colitis-associated cancer . Nonresolving inflammation in the intestine predisposes individuals to the development of colitis-associated cancer ( CAC ) . Inflammasomes are thought to mediate intestinal homeostasis , and their dysregulation contributes to inflammatory bowel diseases and CAC . However , few agents have been reported to reduce CAC by targeting inflammasomes . Here we show that the small molecule andrographolide ( Andro ) protects mice against azoxymethane/dextran sulfate sodium-induced colon carcinogenesis through inhibiting the Q96P20 inflammasome . Administration of Andro significantly attenuated colitis progression and tumor burden . Andro also inhibited Q96P20 inflammasome activation in macrophages both in vivo and in vitro , as indicated by reduced expression of cleaved P29466 , disruption of Q96P20 - Q9ULZ3 - P29466 complex assembly , and lower P01584 secretion . Importantly , Andro was found to trigger mitophagy in macrophages , leading to a reversed mitochondrial membrane potential collapse , which in turn inactivated the Q96P20 inflammasome . Moreover , downregulation of the P42336 - P31749 - P42345 - P23443 pathway accounted for Andro-induced autophagy . Finally , Andro-driven inhibition of the Q96P20 inflammasome and amelioration of murine models for colitis and CAC were significantly blocked by Q14457 knockdown , or by various autophagy inhibitors . Taken together , our findings demonstrate that mitophagy-mediated Q96P20 inflammasome inhibition by Andro is responsible for the prevention of CAC . Our data may help guide decisions regarding the use of Andro in patients with inflammatory bowel diseases , which ultimately reduces the risk of CAC . 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 . 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 . Selective increases of AMPA , DB01221 , and kainate receptor subunit mRNAs in the hippocampus and orbitofrontal cortex but not in prefrontal cortex of human alcoholics . Glutamate is the main excitatory transmitter in the human brain . Drugs that affect the glutamatergic signaling will alter neuronal excitability . DB00898 inhibits glutamate receptors . We examined the expression level of glutamate receptor subunit mRNAs in human post-mortem samples from alcoholics and compared the results to brain samples from control subjects . RNA from hippocampal dentate gyrus ( HP-DG ) , orbitofrontal cortex ( OFC ) , and dorso-lateral prefrontal cortex ( DL- P27918 ) samples from 21 controls and 19 individuals with chronic alcohol dependence were included in the study . Total RNA was assayed using quantitative RT-PCR . Out of the 16 glutamate receptor subunits , mRNAs encoding two AMPA [ 2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid ] receptor subunits P42262 and P42263 ; three kainate receptor subunits Q13002 , Q13003 and Q16478 and five DB01221 ( N-methyl-D-aspartate ) receptor subunits Q05586 , Q12879 , Q14957 , O15399 , and Q8TCU5 were significantly increased in the HP-DG region in alcoholics . In the OFC , mRNA encoding the DB01221 receptor subunit Q8TCU5 was increased , whereas in the DL- P27918 , no differences in mRNA levels were observed . Our laboratory has previously shown that the expression of genes encoding inhibitory GABA-A receptors is altered in the HP-DG and OFC of alcoholics ( Jin et al. , 2011 ) . Whether the changes in one neurotransmitter system drives changes in the other or if they change independently is currently not known . The results demonstrate that excessive long-term alcohol consumption is associated with altered expression of genes encoding glutamate receptors in a brain region-specific manner . It is an intriguing possibility that genetic predisposition to alcoholism may contribute to these gene expression changes . 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 . 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 . 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 . 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 ) . 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 . 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 . The cloning and reintroduction into animal cells of a functional CAD gene , a dominant amplifiable genetic marker . Rodent cells resistant to DB03459 , a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional P27708 , overproduce CAD as a result of amplification of the CAD gene . We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector . Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long . We introduced the cloned genes into CAD-deficient Chinese hamster ovary ( CHO ) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids . We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of DB03459 following protoplast fusion . The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes . The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization . In two of these transformants , the genes are located in one X chromosome or in a chromosome resembling the X . In the third case , the genes are located in a small metacentric or rearranged chromosome . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 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 . 5- DB00233 Inhibits Acute Clostridium difficile Toxin A-Induced Colitis in Rats . We tested the hypothesis that DB00244 ( DB00244 ) inhibits toxin A-induced generation of colonic leukotriene B4 ( LTB4 ) and toxin A colitis in rats . Isolated colonic segments in anesthetized rats were treated intraluminally with toxin A for 3 hours with or without 30 minutes of pretreatment with either DB00244 or sulfapyridine and then colonic tissue levels of LTB4 were measured and inflammation was assessed . Separately , sulfasalazine was administered to rats in their drinking water for 5 days , isolated colonic segments were then prepared , toxin A was administered , and inflammation was assessed as before . Pretreatment with DB00244 inhibited toxin A-induced increased tissue LTB4 concentration in the colon . Sulfasalazine and DB00244 but not sulfapyridine significantly inhibited toxin A colitis . However , pretreatment with DB00244 did not protect against direct Q8NER1 -mediated colitis caused by capsaicin . Toxin A stimulated the release of DB05875 ( SP ) , and this effect was also inhibited by sulfasalazine and DB00244 but not by sulfapyridine . Thus , toxin A stimulates colonic LTB4 resulting in activation of Q8NER1 , release of SP , and colitis . Inhibition of P09917 by DB00244 disrupts this pathway and supports the concept that LTB4 activation of Q8NER1 plays a role in toxin A colitis .

MH_train_1433

MH_train_1433

MH_train_1433

interacts_with DB00877?

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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 . Selectively frequent expression of P32302 enhances resistance to apoptosis in CD8(+) P28906 (+) T cells from patients with T-cell-lineage acute lymphocytic leukemia . We investigated P01730 (+) P28906 (+) , CD8(+) P28906 (+) , P01730 (+) P28906 (-) , and CD8(+) P28906 (-) T cells from cord blood and from typical patients with T-cell-lineage acute lymphocytic leukemia and T-cell-lineage chronic lymphocytic leukemia in terms of expression and functions of P32302 / O43927 . We found that P32302 was selectively frequently expressed on T-cell-lineage acute ( chronic ) lymphocytic leukemia ( T-ALL ) CD8(+) P28906 (+) T cells , but not on T-ALL P01730 (+) P28906 (+) , P01730 (+) P28906 (-) , and CD8(+) P28906 (-) T cells . P32302 was rarely expressed on all types of P28906 (+) and P28906 (-) CB or T-CLL T cells . O43927 /B cells attracting chemokine 1 induced significant resistance to P01375 -mediated apoptosis in T-ALL CD8(+) P28906 (+) T cells , instead of induction of chemotactic and adhesive responsiveness . O75888 expression in T-ALL CD8(+) P28906 (+) T cells was upregulated by O43927 / O43927 ( B-cell attracting chemokine 1 ) . The P32302 / O43927 pair by means of activation of APRIL ( O75888 ) induced resistance to apoptosis in T-ALL CD8(+) P28906 (+) T cells in livin-dependent manner . In this process , cell-cell contact in culture was necessary . Based on our findings , we suggested that there were differential functions of P32302 / O43927 in distinct types of cells . Normal lymphocytes , especially naive B and T cells , utilized P32302 / O43927 for migration , homing , maturation , and cell homeostasis , as well as secondary lymphoid tissue organogenesis . Meanwhile , certain malignant cells took advantages of P32302 / O43927 for infiltration , resistance to apoptosis , and inappropriate proliferation . DB00877 induces Q8NHJ6 (high) Q8N423 (high) dendritic cells promoting a new immunoregulatory pathway . Q8NHJ6 (high) Q8N423 (high) dendritic cells ( DCs ) may cause anergy in P01730 (+)CD45RO(+)CD25(+) T cells transforming them into regulatory T cells ( Tregs ) . Here , we tested whether chronic exposure to rapamycin may modulate this immunoregulatory pathway in renal transplant recipients . Forty renal transplant patients with biopsy-proven chronic allograft nephropathy and receiving calcineurin inhibitors were randomly assigned to either calcineurin inhibitor dose reduction or withdrawal with rapamycin introduction . At conversion and 2 years thereafter , we measured the rapamycin effects on circulating DCs ( BDCA1/ Q8WTT0 and Q8NHJ6 / Q8N423 expression ) , P01730 (+)/CD25(high)/Foxp3(+) Tregs , CD8(+)/ P10747 (-) T cells , and the Th1/Th2 balance in graft biopsies . In rapamycin-treated patients , peripheral Q8WTT0 (+) cells were significantly increased along with Q8NHJ6 / Q8N423 (+) DCs . The number of circulating P01730 (+)/CD25(high)/Foxp3(+)/ P16410 (+) Tregs , CD8(+) P10747 (-) T cells , and P17693 serum levels were higher in the rapamycin-treated group . The number of Q8NHJ6 / Q8N423 (+) Q8WTT0 (+) DC was directly and significantly correlated with circulating Tregs and CD8(+) P10747 (-) T cells . Q8NHJ6 / Q8N423 expression was increased in kidney biopsies at the end of the study period along with a significant bias toward a Th2 response within the graft only in the rapamycin-treated patients . Thus , rapamycin induces the upregulation of Q8NHJ6 and Q8N423 on the DC surface , and this effect is associated with an increase in the number of Tregs and expansion of the CD8(+) P10747 (-) T cell population . This suggests that P42345 inhibition may promote a novel immunoregulatory pathway . Antitumor activity of Triolimus : a novel multidrug-loaded micelle containing Paclitaxel , DB00877 , and 17- P29372 . Triolimus is a first-in-class , multidrug-loaded micelle containing paclitaxel , rapamycin , and 17- P29372 . In this study , we examine the antitumor mechanisms of action , efficacy , and toxicity of Triolimus in vitro and in vivo . In vitro cytotoxicity testing of Triolimus was conducted using two aggressive adenocarcinomas including the lung cancer cell line , A549 , and breast cancer cell line , MDA-MB-231 . The three-drug combination of paclitaxel , rapamycin , and 17- P29372 displayed potent cytotoxic synergy in both A549 and MDA-MB-231 cell lines . Mechanistically , the drug combination inhibited both the Ras/Raf/mitogen-activated protein kinase and PI3K/Akt/ P42345 pathways . Triolimus was advanced into tumor xenograft models for assessment of efficacy , toxicity , and mechanisms of action . In vivo , a three-infusion schedule of Triolimus inhibited A549 and MDA-MB-231 tumor growth far more potently than paclitaxel-containing micelles and effected tumor cures in MDA-MB-231 tumor-bearing animals . Tumor growth delays resulted from a doubling in tumor cell apoptosis and a 50 % reduction in tumor cell proliferation compared with paclitaxel-containing micelles . Enhanced antitumor efficacy was achieved without clinically significant increases in acute toxicity . Thus , Triolimus displays potent synergistic activity in vitro and antitumor activity in vivo with comparable toxicity to paclitaxel . These observations provide strong support for further development of Triolimus and an important proof of concept for safe , effective nanoparticle-based delivery of three complementary anticancer agents . DB00877 antagonizes P01375 induction of P19320 on endothelial cells by inhibiting mTORC2 . Recruitment of circulating leukocytes into inflamed tissues depends on adhesion molecules expressed by endothelial cells ( ECs ) . Here we report that rapamycin pretreatment reduced the ability of P01375 -treated ECs to capture T cells under conditions of venular flow . This functional change was caused by inhibition of P01375 -induced expression of vascular cell adhesion molecule-1 ( P19320 ) and could be mimicked by knockdown of mammalian target of rapamycin ( P42345 ) or rictor , but not raptor , implicating mTORC2 as the target of rapamycin for this effect . Mechanistically , mTORC2 acts through Akt to repress Raf1- Q02750 /2- P27361 /2 signaling , and inhibition of mTORC2 consequently results in hyperactivation of P27361 /2 . Increased P27361 /2 activity antagonizes P19320 expression by repressing P01375 induction of the transcription factor P10914 . Preventing activation of P27361 /2 reduced the ability of rapamycin to inhibit P01375 -induced P19320 expression . In vivo , rapamycin inhibited mTORC2 activity and potentiated activation of P27361 /2 . These changes correlated with reduced endothelial expression of P01375 -induced P19320 , which was restored via pharmacological inhibition of P27361 /2 . Functionally , rapamycin reduced infiltration of leukocytes into renal glomeruli , an effect which was partially reversed by inhibition of P27361 /2 . These data demonstrate a novel mechanism by which rapamycin modulates the ability of vascular endothelium to mediate inflammation and identifies endothelial mTORC2 as a potential therapeutic target . A double-blind , placebo-controlled outpatient trial of pergolide for cocaine dependence . Results of preclinical studies suggest that pergolide , a mixed D(1)/ P14416 agonist , may be useful in treating cocaine dependence . To empirically investigate this possibility , we conducted a 5-year , double-blind , placebo-controlled clinical trial of two doses of pergolide ( 0.05 and 0.25 mg bid ) in subjects with cocaine dependence and combined cocaine/alcohol dependence . Data analysis was performed on an intent to treat population ( N=358 ) and a per protocol population ( N=108 ) with urine drug screens ( UDS ) used as the main outcome measure . There were no significant effects on UDS at either pergolide dose . DB01186 had no significant effect on alcohol use in the comorbid alcohol/cocaine dependence group . DB01186 does not appear to have clinical value in the treatment of cocaine dependence or in decreasing alcohol use in cocaine-dependent individuals at the presently studied doses . A genomic insight into diversity among tribal and nontribal population groups of Manipur , India . Twenty autosomal markers , including linked markers at two gene markers , are used to understand the genomic similarity and diversity among three tribal ( Paite , Thadou , and Kom ) and one nontribal communities of Manipur ( Northeast India ) . Two of the markers ( P01730 and HB9 ) are monomorphic in Paite and one ( the P01730 marker ) in Kom . Data suggest the Meitei ( nontribal groups ) stand apart from the three tribal groups with respect to higher heterozygosity ( 0.366 ) and presence of the highest ancestor haplotypes of P14416 markers ( 0.228 ) ; this is also supported by principal co-ordinate analysis . These populations are found to be genomically closer to the Chinese population than to other Indian populations . 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 . Influence of FcγRIIIA genetic polymorphism on T-lymphocyte depletion induced by rabbit antithymocyte globulins in kidney transplant patients . INTRODUCTION : Polyclonal antithymocyte globulins ( ATG ) have been used in transplantation for several decades , but the sources of the interindividual variability of their effect are poorly understood . An influence of the P08637 -158V/F genetic polymorphism on the horse ATG concentration-effect relationship was reported in kidney transplant patients . The objective of the present study was to confirm the influence of the P08637 polymorphism on the extent of lymphocyte depletion in kidney transplant patients treated with rabbit antithymocyte globulin ( r-ATG ) . MATERIALS AND METHODS : Of the 194 transplant patients treated with r-ATG between 1998 and 2002 in our institution , 69 patients were eligible and included in this retrospective study . Biomarkers of response were CD3 and P01730 counts . Dose-effect data were analyzed using a population approach , and a two-compartment turnover model with stimulation of lymphocyte ' output ' . Since r-ATG concentrations were not available , a K-PD model was used . The influence of P08637 genotype on estimated parameters was investigated . RESULTS : The r-ATG infusion rate leading to a 50 % stimulation of CD3+ output ( EDK(50) ) , which is inversely related to patient sensitivity to r-ATG treatment , decreased with the number of V alleles ( P=0.0016 ) . CONCLUSION : The genetic polymorphism of P08637 influences r-ATG effect on CD3 count in kidney transplant patients , those with the V allele being more sensitive to antilymphocyte serum . These results also suggest that r-ATG act , at least in part , by antibody-dependent cellular cytotoxicity . The combination of rapamycin and MAPK inhibitors enhances the growth inhibitory effect on Nara-H cells . The inhibition of the mammalian target of rapamycin ( P42345 ) signaling pathway promotes the initiation of autophagy , and the mitogen-activated protein kinase ( MAPK ) /extracellular signal-regulated protein kinase ( P29323 ) is well known to induce autophagy . Autophagy is a self-defense mechanism of cancer cells that are subjected to antitumor agents , and blocking autophagy can trigger apoptosis . In the present study , we demonstrate that an P42345 inhibitor , rapamycin , induces autophagy in the Nara-H malignant fibrous histiocytoma ( Q9H334 ) cell line through the activation of P27361 /2 . DB00877 -induced apoptosis was enhanced following the inhibition of the MEK/ P29323 pathway . In the Nara-H cells , we examined the effects of rapamycin treatment on cell proliferation and on the phosphorylation of the P42345 pathway components and autophagy by western blot analysis . Furthermore , we examined the effects of rapamycin with or without the MEK inhibitor , U0126 , on the induction of apoptosis by using fluorescence microscopy . DB00877 inhibited Nara-H cell proliferation and decreased the phosphorylation of the P42345 pathway in the Nara-H cells . DB00877 induced the apoptosis of Nara-H cells , and this apoptosis was enhanced by U0126 . Simultaneously , phospho- P27361 /2 was activated by rapamycin . The present study demonstrates that rapamycin induces autophagy in Nara-H cells by activating the MEK/ P29323 signaling pathway , and the rapamycin-induced apoptosis can be enhanced by the MEK inhibitor , U0126 . These results suggest that self‑protective mechanisms involving P42345 inhibitors in Nara-H cells are prevented by the inhibition of the MEK/ P29323 pathway . The combination of an P42345 inhibitor ( e.g. , rapamycin ) and an MEK inhibitor ( e.g. , U0126 ) may offer effective treatment for Q9H334 , as this combination effectively activates apoptotic pathways . Preclinical in vivo evaluation of rapamycin in human malignant peripheral nerve sheath explant xenograft . Neurofibromatosis type 1 ( P21359 ) patients are prone to the development of malignant tumors , the most common being Malignant Peripheral Nerve Sheath Tumor ( MPNST ) . P21359 -MPNST patients have an overall poor survival due to systemic metastasis . Currently , the management of MPNSTs includes surgery and radiation ; however , conventional chemotherapy is not very effective , underscoring the need for effective biologically-targeted therapies . Recently , the P21359 gene product , neurofibromin , was shown to negatively regulate the phosphoinositide-3-kinase ( PI3K ) /Protein Kinase-B ( Akt ) /mammalian Target Of DB00877 ( P42345 ) pathway , with loss of neurofibromin expression in established human MPNST cell lines associated with high levels of P42345 activity . We developed and characterized a human P21359 -MPNST explant grown subcutaneously in NOD-SCID mice , to evaluate the effect of the P42345 inhibitor rapamycin . We demonstrate that rapamycin significantly inhibited human P21359 -MPNST P42345 pathway activation and explant growth in vivo at doses as low as 1.0 mg/kg/day , without systemic toxicities . While rapamycin was effective at reducing P21359 -MPNST proliferation and angiogenesis , with decreased CyclinD1 and P15692 respectively , there was no increase in tumor apoptosis . DB00877 effectively decreased activation of S6 downstream of P42345 , but there was accompanied increased Akt activation . This study demonstrates the therapeutic potential and limitations of rapamycin in P21359 -associated , and likely sporadic , MPNSTs . Q13972 (Mm)/ Q13972 regulates both Ras and Rac signaling pathways . The Q13972 exchange factor molecule contains in addition to the catalytic domain two pleckstrin homology ( P78364 and Q8IXK0 ) , one IQ and one Dbl homology ( DH ) domains . In this study we investigated the role of such additional domains . We found that a Q13972 mutant lacking P78364 and IQ domains is sufficient to activate c-fos promoter in response to lysophosphatidic acid ( P08519 ) . The same mutant did not increase external stimuli-regulated kinase ( P29323 ) activity , suggesting an additional mechanism for the induction of gene transcription . Isolated DH- Q8IXK0 module activates c-Jun NH(2)-terminal kinase and the c-fos promoter in response to P08519 , providing the basis for an P29323 -independent mechanism . These results provide evidence that Q13972 acts as a bifunctional molecule on both P29323 -dependent and independent pathways . DB05578 , a fully human mAb to the transmembrane signaling tyrosine kinase P35968 for the potential treatment of cancer . Angiogenesis is essential for tumor growth , invasion and metastasis , and is mediated , at least in part , by a large family of P15692 ligands and receptors . DB05578 , which is being developed by ImClone Systems Inc , is a fully human mAb that binds human P35968 , thus blocking P15692 binding and inhibiting angiogenesis . Proof-of-concept preclinical studies with the mouse mAb DC-101 supported this hypothesis , and ramucirumab inhibited cell proliferation in vitro , as well as tumor progression in mouse xenograft models of human cancer . DB05578 was well tolerated on weekly and fortnightly schedules in phase I clinical trials in patients with advanced cancers ; mechanism-related DLTs were hypertension and deep venous thrombosis . Stable disease was also observed in several patients treated on either schedule , and several patients on the weekly schedule exhibited partial responses . At the time of publication , ramucirumab was undergoing assessment in phase II trials as a monotherapy in hepatocellular , renal cell and ovarian carcinomas . DB05578 was also in phase II trials in combination with dacarbazine in melanoma , with mitoxantrone/prednisone in prostate cancer , with carboplatin/paclitaxel in NSCLC and with oxaliplatin/folinic acid/5-fluorouracil in colorectal cancer . A phase III trial in combination with docetaxel in breast cancer was also ongoing . Pending results from these trials , ramucirumab may be a useful addition to current antiangiogenic therapies . The results are awaited with interest . Interactome mapping of the phosphatidylinositol 3-kinase-mammalian target of rapamycin pathway identifies deformed epidermal autoregulatory factor-1 as a new glycogen synthase kinase-3 interactor . The phosphatidylinositol 3-kinase-mammalian target of rapamycin ( PI3K- P42345 ) pathway plays pivotal roles in cell survival , growth , and proliferation downstream of growth factors . Its perturbations are associated with cancer progression , type 2 diabetes , and neurological disorders . To better understand the mechanisms of action and regulation of this pathway , we initiated a large scale yeast two-hybrid screen for 33 components of the PI3K- P42345 pathway . Identification of 67 new interactions was followed by validation by co-affinity purification and exhaustive literature curation of existing information . We provide a nearly complete , functionally annotated interactome of 802 interactions for the PI3K- P42345 pathway . Our screen revealed a predominant place for glycogen synthase kinase-3 ( GSK3 ) A and B and the AMP-activated protein kinase . In particular , we identified the deformed epidermal autoregulatory factor-1 ( O75398 ) transcription factor as an interactor and in vitro substrate of P49840 and P49841 . Moreover , GSK3 inhibitors increased O75398 transcriptional activity on the P08908 serotonin receptor promoter . We propose that O75398 may represent a therapeutic target of lithium and other GSK3 inhibitors used in bipolar disease and depression . Oxanine DNA glycosylase activity from Mammalian alkyladenine glycosylase . Oxanine ( Oxa ) is a deaminated base lesion derived from guanine in which the N(1)-nitrogen is substituted by oxygen . This work reports the mutagenicity of oxanine as well as oxanine DNA glycosylase ( ODG ) activities in mammalian systems . Using human P06746 , deoxyoxanosine triphosphate is only incorporated opposite cytosine ( Cyt ) . When an oxanine base is in a DNA template , Cyt is efficiently incorporated opposite the template oxanine ; however , adenine and thymine are also incorporated opposite Oxa with an efficiency approximately 80 % of a Cyt/Oxa ( C/O ) base pair . DB02377 is incorporated opposite Oxa with the least efficiency , 16 % compared with cytosine . ODG activity was detected in several mammalian cell extracts . Among the known human DNA glycosylases tested , human alkyladenine glycosylase ( P29372 ) shows ODG activity , whereas hOGG1 , hNEIL1 , or hNEIL2 did not . ODG activity was detected in spleen cell extracts of wild type age-matched mice , but little activity was observed in that of Aag knock-out mice , confirming that the ODG activity is intrinsic to P29372 . Human P29372 can excise Oxa from all four Oxa-containing double-stranded base pairs , Cyt/Oxa , Thy/Oxa , DB00173 /Oxa , and Gua/Oxa , with no preference to base pairing . Surprisingly , P29372 can remove Oxa from single-stranded Oxa-containing DNA as well . Indeed , P29372 can also remove 1,N(6)-ethenoadenine from single-stranded DNA . This study extends the deaminated base glycosylase activities of P29372 to oxanine ; thus , P29372 is a mammalian enzyme that can act on all three purine deamination bases , hypoxanthine , xanthine , and oxanine . Primary hyperoxaluria type I due to a point mutation of T to C in the coding region of the serine:pyruvate aminotransferase gene . cDNA clones for serine:pyruvate aminotransferase ( P21549 , alternative name : alanine:glyoxylate aminotransferase ) were obtained from a cDNA library constructed from the liver of a primary hyperoxaluria type I ( P78364 ) case in which the P21549 activity was approximately one-hundredth that in control liver . Six clones were isolated from 100,000 transformants and all of them contained an approximately 1.5 kbp insert which included the whole coding region for human P21549 . Nucleotide sequence analysis revealed a point mutation of T to C at position 634 ( relative to the 5'-end of the cDNA ) encoding a DB00133 to Pro substitution at residue 205 . The T to C conversion created a new SmaI site , which enabled us to demonstrate that the point mutation had occurred in the patient 's P21549 gene . SmaI digestion of genomic DNA may be useful for the diagnostic gene analysis of this type of P78364 . 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 . P62937 and nuclear factor of activated T cells are essential in cyclosporine-mediated suppression of polyomavirus BK replication . Immunosuppressants have impacts on the development of polyomavirus-associated nephropathy . We previously demonstrated that cyclosporin A ( DB00091 ) suppressed polyomavirus BK ( BKV ) replication . The role of cyclophilin A ( CypA ) and nuclear factor of activated T cells ( NFAT ) in DB00091 -imposed suppression of BKV replication was determined in this study . Results demonstrated that knockdown of CypA but not CypB significantly reduced BKV large T antigen ( TAg ) expression and BKV titer . Overexpression of CypA reversed CypA siRNA-induced inhibition in BKV TAg expression . In addition , CypA overexpression attenuated the suppressive effect of DB00091 on TAg expression , suggesting CypA implicated in DB00091 -mediated anti-BKV effect . Knockdown of Q12968 abrogated TAg expression , while overexpression of Q12968 promoted TAg expression and augmented BKV promoter activity . Q12968 binding to the BKV promoter was verified by chromatin immunoprecipitation assay and electrophoretic mobility shift assay . Renal histology also displayed an increase in Q12968 expression in tubulointerstitium of BKV-associated nephropathy . Furthermore , overexpression of Q12968 rescued DB00091 -mediated inhibition of BKV load and TAg expression . A DB00091 analog , NIM811 , which can not block NFAT functionality , failed to suppress TAg expression . In conclusion , CypA and NFAT are indispensable in BKV replication . DB00091 inhibits BKV replication through CypA and NFAT , which may be potential targets of anti-BKV treatment . 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 . Role of antispasmodics in the treatment of irritable bowel syndrome . Irritable bowel syndrome ( IBS ) is a long-lasting , relapsing disorder characterized by abdominal pain/discomfort and altered bowel habits . Intestinal motility impairment and visceral hypersensitivity are the key factors among its multifactorial pathogenesis , both of which require effective treatment . Voltage-gated calcium channels mediate smooth muscle contraction and endocrine secretion and play important roles in neuronal transmission . Antispasmodics are a group of drugs that have been used in the treatment of IBS for decades . DB01616 citrate , a spasmolytic , decreases the sensitivity of smooth muscle contractile proteins to calcium , and it is a selective P08908 receptor antagonist . DB01616 , in combination with simethicone , has been demonstrated to effectively reduce abdominal pain and discomfort in a large placebo-controlled trial . Mebeverine is a musculotropic agent that potently blocks intestinal peristalsis . Non-placebo-controlled trials have shown positive effects of mebeverine in IBS regarding symptom control ; nevertheless , in recent placebo-controlled studies , mebeverine did not exhibit superiority over placebo . Otilonium bromide is poorly absorbed from the GI tract , where it acts locally as an L-type calcium channel blocker , an antimuscarinic and a tachykinin NK2 receptor antagonist . Otilonium has effectively reduced pain and improved defecation alterations in placebo-controlled trials in IBS patients . DB09090 bromide is also an L-type calcium channel blocker that acts locally in the GI tract . DB09090 improves motility disorders and consequently reduces stool problems in IBS patients . Phloroglucinol and trimethylphloroglucinol are non-specific antispasmodics that reduced pain in IBS patients in a placebo-controlled trial . Antispasmodics have excellent safety profiles . T-type calcium channel blockers can abolish visceral hypersensitivity in animal models , which makes them potential candidates for the development of novel therapeutic agents in the treatment of IBS . Proof of concept study to assess fetal gene expression in amniotic fluid by nanoarray PCR . Microarray analysis of cell-free RNA in amniotic fluid ( AF ) supernatant has revealed differential fetal gene expression as a function of gestational age and karyotype . Once informative genes are identified , research moves to a more focused platform such as quantitative reverse transcriptase-PCR . Standardized NanoArray PCR ( P60880 ) is a recently developed gene profiling technology that enables the measurement of transcripts from samples containing reduced quantities or degraded nucleic acids . We used a previously developed P60880 gene panel as proof of concept to determine whether fetal functional gene expression could be ascertained from AF supernatant . RNA was extracted and converted to cDNA from 19 AF supernatant samples of euploid fetuses between 15 to 20 weeks of gestation , and transcript abundance of 21 genes was measured . Statistically significant differences in expression , as a function of advancing gestational age , were observed for 5 of 21 genes . P08758 , P08236 , and P62937 showed decreasing gene expression over time , whereas O15234 and O43296 showed increasing gene expression over time . Statistically significantly increased expression of P42345 and P52630 was seen in female compared with male fetuses . This study demonstrates the feasibility of focused fetal gene expression analysis using P60880 technology . In the future , this technique could be optimized to examine specific genes instrumental in fetal organ system function , which could be a useful addition to prenatal care . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually .

MH_train_1434

MH_train_1434

MH_train_1434

interacts_with DB00502?

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 . 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 . 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 . Abnormal sensitivity of cortisol-producing adrenocortical adenomas to serotonin : in vivo and in vitro studies . Two patients with incidentally discovered adrenocortical adenomas underwent a series of pharmacological and physiological tests after pretreatment with dexamethasone . Illicit plasma cortisol responses to the serotonin (5-HT)4 receptor agonist cisapride were observed in the two patients . Significant increases in plasma cortisol levels were also noticed after glucagon and combined TRH/ DB00644 / P01286 stimulation tests in patient 1 and after administration of the lysine vasopressin precursor terlipressin in patient 2 . After adrenalectomy , in vitro studies were conducted to investigate the cortisol responses of cultured tumor cells to serotonergic ligands and peptide hormones . In the two cases , 5-HT stimulated cortisol secretion from tumor cells with increased efficacy and/or potency to activate steroidogenesis by comparison with normal adrenocortical cells . The corticotropic effect of 5-HT was inhibited by the specific Q13639 receptor antagonist GR 113808 and more potently by methiothepin , a nonspecific serotonergic antagonist having no affinity for the Q13639 receptor . These results show that the hypersensitivity of the tumors to 5-HT was related to tissue expression of an ectopic serotonergic receptor in addition to the eutopic Q13639 receptor . In the two adenoma tissues , immunohistochemical studies revealed the presence of 5-HT-like immunoreactivity within clusters of steroidogenic cells , suggesting that 5-HT acted through an autocrine/paracrine mechanism to stimulate steroidogenesis . Glucagon and DB00644 but not TRH , P01286 , and human chorionic gonadotropin stimulated cortisol secretion from tumor 1 cells . In conclusion , this study provides the first observation of adrenocortical cortisol-producing adenomas hypersensitive in vivo and in vitro to serotonergic agonists . Our results also show that cortisol-producing adenomas can express simultaneously several illegitimate receptors . 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 . 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 . 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 . Subacute lupus erythematosus during treatment with DB06674 for seronegative rheumatoid arthritis . We report on a 52-year-old woman with a history of severe seronegative rheumatoid arthritis . Several conventional therapies and biological therapy with etanercept and infliximab had been unsuccessful . In 2010 she was given DB06674 subcutaneously at a monthly dose of 50 mg . She had a negative Q14201 titre . After 16 months of uninterrupted therapy and sustained response , she developed skin lesions on the upper trunk , back and upper extremities , which worsened on exposure to the sun . The skin biopsy was compatible with subacute lupus erythematosus . Laboratory findings included an Q14201 titre 1:640 , negative anti-Ro/SSA and anti-DNA antibodies . Topical corticosteroid therapy proved inadequate . The patient 's condition improved only after discontinuation of DB06674 . The causal relationship between subacute cutaneous lupus erythematosus and DB06674 is not dose-related and occurs with some delay ( a typical feature of immunological adverse reactions ) . The association is likely , but not confirmed ( because re-challenge was not performed ) . However , a clear improvement was noted after withdrawal . Based on this case , we hypothesized the aetiological role of DB06674 -associated immunogenicity . P01375 -α antagonist-induced lupus-like syndrome ( TAILS ) is a well-known side effect of this class of substances . The British Society of Rheumatology recommends discontinuation of the causal anti- P01375 -α treatment in patients with TAILS . 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 . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . [ Melanoma : from molecular studies to the treatment breakthrough ] . Melanoma holds a leading position in the mortality from skin tumors . Standard treatment of metastatic melanoma allows tumor remission to be achieved only in a small subset of patients . Studies on melanoma molecular pathogenesis led to the identification of several causative genetic events and , consequently , to the development of novel targeted drugs . More than a half of melanomas contain amine acid substitutions in serine-threonine kinase P15056 . Clinical trials involving specific P15056 inhibitors -- vemurafenib and dabrafenib -- demonstrated high efficacy of these agents towards P15056 -mutated melanoma . MEK inhibitors may show activity against both P15056 -- and P01111 -driven tumors . Mucosal and acral melanomas frequently contain mutation in P10721 receptor and can be successfully treated by imatinib . There are novel therapeutic monoclonal antibodies targeted against immunosuppressive molecules P16410 , P18621 and Q9NZQ7 . In some instances these drugs allow to obtain exceptionally prolonged responses . Whole genome sequencing led to the identification of new melanoma genes , e.g. Q12879 , Q9Y4A5 , Q70Z35 , P63000 , P49842 , O00743 , etc . Molecular testing , especially P15056 mutation analysis , has become a mandatory part of melanoma diagnosis . Nevertheless , despite the revolution in melanoma treatment , the prevention of excessive ultraviolet exposure , cancer awareness and early diagnosis remain the main tools for the management of this disease . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . The novel DB01221 receptor antagonist , 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino)-benzoic acid , is a gating modifier in cultured mouse cortical neurons . Neu2000 [ P04626 , 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino)-benzoic acid ] , a derivative of sulfasalazine , attenuates DB01221 -induced neuronal toxicity . Here we investigated the effects of P04626 on the DB01221 receptor ( NMDAR ) using whole-cell patch clamp technique to determine the molecular mechanisms underlying its neuroprotective role . P04626 reversibly suppressed DB01221 responses in an uncompetitive manner with fast binding kinetics . Its inhibition of NMDAR activity depended on both the concentration and the use of agonist but not on the membrane potential . P04626 accelerated DB01221 desensitization without affecting the binding affinity of NMDAR for its agonists and stabilized the closed state of NMDAR . Therefore , P04626 should effectively alleviate disorders that are a result of glutamate excitoxicity with fewer side effects because it is a low-affinity gating modifier that antagonizes NMDAR in an uncompetitive manner . Moreover , in the presence of ifenprodil ( an Q13224 antagonist ) but not DB00238 -AAM077 [ ( R ) - [ ( S ) -1-(4-bromo-phenyl)-ethylamino ] -(2,3-dioxo-1,2,3,4-tetrahydro-quinoxalin-5-yl)-methyl ] -phosphonic acid , an Q12879 antagonist ] , the extent of P04626 block was decreased , suggesting that P04626 is an Q13224 -specific antagonist . CaMKII autonomy is substrate-dependent and further stimulated by Ca2+/calmodulin . A hallmark feature of Ca(2+)/calmodulin ( P62158 ) -dependent protein kinase II ( CaMKII ) regulation is the generation of Ca(2+)-independent autonomous activity by DB00156 -286 autophosphorylation . CaMKII autonomy has been regarded a form of molecular memory and is indeed important in neuronal plasticity and learning/memory . DB00156 -286-phosphorylated CaMKII is thought to be essentially fully active ( approximately 70-100 % ) , implicating that it is no longer regulated and that its dramatically increased Ca(2+)/ P62158 affinity is of minor functional importance . However , this study shows that autonomy greater than 15-25 % was the exception , not the rule , and required a special mechanism ( T-site binding ; by the T-substrates AC2 or Q13224 ) . Autonomous activity toward regular R-substrates ( including tyrosine hydroxylase and GluR1 ) was significantly further stimulated by Ca(2+)/ P62158 , both in vitro and within cells . Altered K(m) and V(max) made autonomy also substrate- ( and DB00171 ) concentration-dependent , but only over a narrow range , with remarkable stability at physiological concentrations . Such regulation still allows molecular memory of previous Ca(2+) signals , but prevents complete uncoupling from subsequent cellular stimulation . DB00072 emtansine in human epidermal growth factor receptor 2-positive breast cancer : a review . PURPOSE OF REVIEW : In this review , we aim to update the clinical data of trastuzumab-DM1 ( DB05773 ) in terms of safety and efficacy , and describe ongoing and future trials evaluating its potential role in the management of patients with human epidermal growth factor receptor 2 ( P04626 ) -positive breast cancer . RECENT FINDINGS : DB00072 emtansine ( DB05773 ) is an antibody drug conjugate that optimizes delivery of chemotherapy with an anti- P04626 monoclonal antibody . As a conjugate , DB05773 's systemic side effects are significantly minimized due to its targeted delivery by trastuzumab to P04626 -positive cells . Phase I and II studies show that the maximum tolerated dose , and thus the recommended dose for DB05773 , is 3.6 mg/kg given intravenously every 3 weeks . Single arm phase Ib/II , II and a randomized phase II first-line study of DB05773 versus the combination of trastuzumab + docetaxel all showed improved tolerability , and at least equivalent efficacy , compared with our current standard of care . Two randomized phase III registration studies are now active , evaluating this agent in the refractory and first-line P04626 -positive settings . SUMMARY : DB05773 has been shown to be a very promising agent for the targeted delivery of chemotherapy and anti- P04626 monoclonal antibody therapy for patients with metastatic , P04626 -positive breast cancer . DB05773 will likely play a role in the management of patients with advanced and early stage P04626 -positive breast cancer , but this awaits further study . Polymorphisms influencing olanzapine metabolism and adverse effects in healthy subjects . OBJECTIVE : The pharmacokinetics of olanzapine and response to treatment could be affected by polymorphisms in genes coding for drug-metabolizing enzymes , transporters , or receptors . The aim of this study was to identify genetic markers predictive of pharmacokinetics , pharmacodynamics , and adverse effects of olanzapine . METHODS : Sixty-three healthy volunteers receiving a single 5-mg oral dose of olanzapine were genotyped for 39 genetic variants that could be related to the response to olanzapine . All genetic variants were analyzed by PharmaChip , but P14416 Taq1A polymorphism was determined by real-time polymerase chain reaction . Olanzapine was measured using high-performance liquid chromatography combined with tandem mass spectrometry . The relationship of gender and polymorphisms with olanzapine pharmacokinetics , the change in prolactin levels , and the incidence of adverse effects were evaluated by multiple regression analysis . RESULTS : The pharmacokinetics of olanzapine was influenced by polymorphisms in P20815 , P21266 , and Q13224 . P01236 levels were affected by gender and polymorphisms in P14416 and 5- P28223 . Polymorphisms in P11712 , P51580 , P22309 , P08183 , and 5- P28223 were related to some adverse effects of olanzapine . CONCLUSIONS : Several polymorphisms can explain differences in the pharmacokinetics , pharmacodynamics , and safety of olanzapine in healthy subjects . Whether these genetic factors influence the risk of therapeutic failure or tolerability in patients remains to be established . DB00184 -associated cues maintain nicotine-seeking behavior in rats several weeks after nicotine withdrawal : reversal by the cannabinoid ( P21554 ) receptor antagonist , rimonabant ( SR141716 ) . Conditioned stimuli are important for nicotine dependence and may trigger craving and relapse after prolonged nicotine abstinence . However , little is known about the pharmacology of this process . Among the systems that have been shown to play a role in drug-seeking behavior is the endocannabinoid transmission . Therefore , the present study examined the resistance to extinction of drug-seeking behavior elicited by nicotine-associated environmental stimuli and the effects of the selective P21554 cannabinoid antagonist rimonabant ( SR141716 ) on the reinforcing effects of nicotine-related stimuli . Rats were trained to self-administer nicotine ( 0.03 mg/kg/injection , i.v. ) under conditions in which responding was reinforced jointly by response-contingent nicotine injections and stimuli ( light and tone ) . After self-administration acquisition , nicotine was withdrawn and lever pressing was only reinforced by contingent presentation of the audiovisual stimuli . Under such a condition , responding persisted for 3 months , following which nonpresentation of the cues produced a progressive extinction of responding . As expected , rats trained to lever-press for saline injections paired with the audiovisual stimuli did not acquire the self-administration . These findings indicate that the cues required learned association with nicotine to acquire reinforcing properties and to function as conditioned reinforcers . When administered 1 month following nicotine withdrawal , rimonabant ( 1 mg/kg , i.p. ) decreased conditioned behavior . These results showing the persistence of a nicotine-conditioned behavior are congruent with the role of nicotine-related environmental stimuli in nicotine craving in abstinent smokers . DB06155 , which has been shown previously to reduce nicotine self-administration , may be effective not only as an aid for smoking cessation but also in the maintenance of abstinence . 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 ) . Multiple colon perforation as a fatal complication during treatment of metastatic melanoma with ipilimumab - case report . DB06186 , an anticancer drug , is an anti- P16410 monoclonal antibody . It is used in treatment of disseminated melanoma . Therapy is associated with high risk of complications . One of the most serious , although one of the rarest is perforation of gastrointestinal tract . In this case report we describe a 52-year old male , with disseminated melanoma with unknown starting point , treated with anti- P16410 monoclonal antibody . After 3rd dose of drug administration , bloody diarrhea and acute abdominal pain occurred as a symptom of gastrointestinal perforation . A single perforation was sutured during laparotomy . Symptoms of acute abdomen returned after 10 days . Pus-faecalperitonitis , symptoms of necro-hemorrhagic colitis and multilocal perforation of the colon were found during relaparotomy . Pancolectomy with end ileostomy was performed . Few hours since relaparotomy pacient died due to multiple organ failure . The purpose of this case report is to draw attention to a risk of multilocal colon perforation in patient treated with ipilumumab .

MH_train_1435

MH_train_1435

MH_train_1435

interacts_with DB06271?

multiple_choice

Surface expression of P48058 AMPA receptor is dependent on an interaction between its C-terminal domain and a 4.1 protein . Dynamic regulation of the number and activity of AMPA receptors is believed to underlie many forms of synaptic plasticity and is presumably mediated by specific protein-protein interactions involving the C-terminal domain of the receptor . Several proteins interacting with the C-terminal tails of the glutamate receptor ( GluR ) -A and P42262 subunits have been identified and implicated in the regulation of endocytosis and exocytosis , clustering , and anchoring of AMPA receptors to the cytoskeleton . In contrast , little is known of the molecular interactions of the P48058 subunit , or of the mechanisms regulating the traffic of P48058 -containing AMPA receptors . We analyzed the subcellular localization of homomeric P48058 receptors carrying C-terminal deletions in transfected human embryonic kidney ( P29320 ) 293 cells and in primary neurons by immunofluorescence microscopy and ELISA . A minimal requirement for a 14-residue cytoplasmic segment for the surface expression of homomeric P48058 receptors was identified . Previously , a similar region in the P42261 subunit was implicated in an interaction with 4.1 family proteins . Coimmunoprecipitation demonstrated that P48058 associated with 4.1 protein(s) in both HEK293 cells and rat brain . Moreover , glutathione S-transferase pull-down experiments showed that the same 14-residue segment is critical for 4.1 binding to P42261 and P48058 . Point mutations within this segment dramatically decreased the surface expression of P48058 in HEK293 cells , with a concomitant loss of the 4.1 interaction . Our findings demonstrate a novel molecular interaction for the P48058 subunit and suggest that the association with the 4.1 family protein(s) plays an essential role in the transport to and stabilization of P48058 -containing AMPA receptors at the cell surface . All-trans retinoic acid inhibits the increases in fibronectin and P05121 induced by TGF-beta1 and Ang II in rat mesangial cells . AIM : To investigate the effect of all-trans RA ( atRA ) on the increases in plasminogen activator inhibitor-1 ( P05121 ) and fibronectin that are induced by transforming growth factor-beta1 ( TGF-beta1 ) and angiotensin II ( Ang II ) in cultured rat glomerular mesangial cells . METHODS : Subconfluent glomerular mesangial cells were serum-starved for 48 h and pretreated with atRA with subsequent stimulation of TGF-beta1 and Ang II . Protein expressions of cell-associated fibronectin and P05121 in glomerular mesangial cells were evaluated by Western blot analysis . mRNA expression of RA receptors in glomerular mesangial cells was examined by RT-PCR . RESULTS : Retinoic acid receptor-alpha , -gamma ( P10276 , -gamma ) and retinoid X receptor-alpha , -beta , -gamma ( RXR-alpha , -beta , -gamma ) mRNA were expressed in rat glomerular mesangial cells . atRA pretreatment effectively reduced fibronectin expression in glomerular mesangial cells stimulated with TGF-beta 1 or Ang II for 48 h . TGF-beta 1 stimulated P05121 expression reached a maximum at 5 h . atRA did n't affect the early ( 5 h ) P05121 induction by TGF-beta 1 , but markedly attenuated the sustained ( 48 h ) P05121 induction . atRA also decreased the prolonged effect of Ang II on P05121 expression . CONCLUSION : These results indicate that atRA inhibits the increases in fibronectin that are induced by TGF-beta1 and Ang II in cultured glomerular mesangial cells . The data also suggest that this effect of atRA is associated with a change in P05121 levels . Blood coagulation factors changes during liver regeneration in rats . Effects of partial hepatectomy on blood coagulation factors were investigated in rats . Analysis were performed 24 , 48 and 72 hours after surgery . Howell 's time was significantly higher after 24 and 48 h compared to the control value . P00734 time was significantly prolonged after 24 h . Partial thromboplastin time did not differ significantly in any time . FII values were significantly reduced after 24 and 48 h , but FV values only after 24 h . FVII showed significant decrease after 24 h , but significant increase at 48 h . FVIII and P01008 average values were significantly lower after 24 , 48 and 72 h . Plasma fibrinogen increased . Significant differences were observed 48 and 72 h after surgery . Differences in normalization time of these coagulation factors are most probably the consequence of their synthesis in various cell types , regenerated at different periods after partial hepatectomy . Expression and prognostic relevance of activated extracellular-regulated kinases ( P27361 /2 ) in breast cancer . Extracellular-regulated kinases ( P27361 , P28482 ) play important roles in the malignant behaviour of breast cancer cells in vitro . In our present study , 148 clinical breast cancer samples ( 120 cases with follow-up data ) were studied for the expression of P27361 , P28482 and their phosphorylated forms p- P27361 and p- P28482 by immunoblotting , and p- P27361 /2 expression in corresponding paraffin sections was analysed by immunohistochemistry . The results were correlated with established clinical and histological prognostic parameters , follow-up data and expression of seven cell-cycle regulatory proteins as well as P03956 , P14780 , P05121 and AP-1 transcription factors , which had been analysed before . High p- P27361 expression as determined by immunoblots correlated significantly with a low frequency of recurrences and infrequent fatal outcome ( P = 0.007 and 0.008 ) and was an independent indicator of long relapse-free and overall survival in multivariate analysis . By immunohistochemistry , strong p- P29323 staining in tumour cells was associated with early stages ( P = 0.020 ) , negative nodal status ( P = 0.003 ) and long recurrence-free survival ( P = 0.017 ) . In contrast , expression of the unphosphorylated kinases P27361 and P28482 was not associated with clinical and histological prognostic parameters , except a positive correlation with oestrogen receptor status . Comparison with the expression of formerly analysed cell-cycle- and invasion-associated proteins corroborates our conclusion that activation of P27361 and P28482 is not associated with enhanced proliferation and invasion of mammary carcinomas . Isolation , partial purification and characterization of nuclear retinoic acid receptors from chick skin . Nuclear receptors ( RARs ) for retinoic acid ( RA ) are considered to be the ultimate mediators of the action of RA in the control of cell differentiation and inhibition of tumorigenesis . We have isolated and partially purified and characterized RAR from a RA-responsive tissue , chick embryo skin . The purification steps included Affi-Gel blue chromatography , ultrafiltration , size exclusion chromatography , and preparative isoelectric focusing . The electrofocusing of RAR-[3H]RA complex in ampholines ( pH 3-10 ) revealed that the receptors have an isoelectric pH of 7.5 . Whereas pronase-digested the RAR-[3H]RA complex completely , DNase showed 20-35 % and RNase showed negligible digestive action on the complex . The ligand binding to RAR was completely inhibited by a mercury compound . P10276 - and P10826 -specific antibodies , on Western blot analysis , immunoreacted with a protein having a molecular weight of 50,000 , presumably RAR . Binding affinity studies revealed that biologically active analogs of RA with a free COOH group ( e.g. , 13- DB00982 , RO-13-7410 , Ch 55 , and DB04942 ) showed , like RA , high binding affinity for RAR , whereas biologically ineffective analogs of RA ( e.g. , furyl and pyridyl ) were poor binders . Other groups of retinoids , in which the COOH group was either lacking or blocked , did not bind to RAR whether or not they were biologically active . 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 . The contribution of cyclooxygenase-1 and -2 to persistent thromboxane biosynthesis in aspirin-treated essential thrombocythemia : implications for antiplatelet therapy . We tested whether cyclooxygenase 2 ( P35354 ) expression and unacetylated P23219 in newly formed platelets might contribute to persistent thromboxane ( TX ) biosynthesis in aspirin-treated essential thrombocythemia ( ET ) . Forty-one patients on chronic aspirin ( 100 mg/day ) and 24 healthy subjects were studied . Platelet P35354 expression was significantly increased in patients and correlated with thiazole orange-positive platelets ( r = 0.71 , P < .001 ) . The rate of TXA(2) biosynthesis in vivo , as reflected by urinary 11-dehydro-TXB(2) ( TXM ) excretion , and the maximal biosynthetic capacity of platelets , as reflected by serum TXB(2) , were higher in patients compared with aspirin-treated healthy volunteers . Serum TXB(2) was significantly reduced by the selective P35354 inhibitor NS-398 added in vitro . Patients were randomized to adding the selective P35354 inhibitor , etoricoxib , or continuing aspirin for 7 days . DB01628 significantly reduced by approximately 25 % TXM excretion and serum TXB(2) . Fourteen of the 41 patients were studied again 21 ( +/- 7 ) months after the first visit . Serum TXB(2) was consistently reduced by approximately 30 % by adding NS398 in vitro , while it was completely suppressed with 50 microM aspirin . Accelerated platelet regeneration in most aspirin-treated ET patients may explain aspirin-persistent TXA(2) biosynthesis through enhanced P35354 activity and faster renewal of unacetylated P23219 . These findings may help in reassessing the optimal antiplatelet strategy in ET . Intestinal myofibroblast-specific Tpl2-Cox-2- DB00917 pathway links innate sensing to epithelial homeostasis . Tumor progression locus-2 ( Tpl2 ) kinase is a major inflammatory mediator in immune cell types recently found to be genetically associated with inflammatory bowel diseases ( IBDs ) . Here we show that Tpl2 may exert a dominant homeostatic rather than inflammatory function in the intestine mediated specifically by subepithelial intestinal myofibroblasts ( IMFs ) . Mice with complete or IMF-specific Tpl2 ablation are highly susceptible to epithelial injury-induced colitis showing impaired compensatory proliferation in crypts and extensive ulcerations without significant changes in inflammatory responses . Following epithelial injury , IMFs sense innate or inflammatory signals and activate , via Tpl2 , the cyclooxygenase-2 ( Cox-2 ) -prostaglandin E2 ( DB00917 ) pathway , which we show here to be essential for the epithelial homeostatic response . Exogenous DB00917 administration rescues mice with complete or IMF-specific Tpl2 ablation from defects in crypt function and susceptibility to colitis . We also show that Tpl2 expression is decreased in IMFs isolated from the inflamed ileum of Q9UKU7 patients indicating that Tpl2 function in IMFs may be highly relevant to human disease . The IMF-mediated mechanism we propose also involves the Q9UKU7 -associated genes P14778 , P28482 , and the DB00917 receptor-encoding P35408 . Our results establish a previously unidentified myofibroblast-specific innate pathway that regulates intestinal homeostasis and may underlie Q9UKU7 susceptibility in humans . 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 . 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 . Site-specific mutagenesis of the histidine precursor of diphthamide in the human elongation factor-2 gene confers resistance to diphtheria toxin . Protein synthesis elongation factor 2 ( P13639 ) from eukaryotes contains a conserved post-translationally modified histidine residue known as diphthamide . DB03223 is a unique site of ADP-ribosylation by diphtheria toxin ( DT ) , which is responsible for cell killing . In this report , we describe the construction of DT-resistant HeLa cell lines by engineering the toxin-resistant form of its specific substrate , protein elongation factor-2 . Using site-specific mutagenesis of the histidine precursor of diphthamide , the histidine residue of codon 715 in human P13639 cDNA was substituted with one of four amino acid residue codons : leucine , methionine , asparagine or glutamine . Mutant EF-2s were subcloned into a pCMVexSVneo expression vector , transfected into HeLa cells , and DT-resistant cell clones were isolated . The protective effect of mutant EF-2s against cell killing by DT , after exposing all four mutant strains derived from HeLa cells to different concentrations of the toxin ( 5-20 ng/mL ) was demonstrated by : ( 1 ) the normal morphological appearance of the cells ; ( 2 ) their unaffected or slightly slower growth rates ; ( 3 ) their undisturbed electrophoretic DNA profiles whose integrity was virtually preserved . Mutant cell strains showed also considerable levels of resistance to very high concentrations of DT , in that they maintained slower but consistent rates of cell growth . It was hence concluded that despite its strict conservation and unique modification , the diphthamide histidine appears not to be essential to the function of human P13639 in protein synthesis . In addition , DT-resistant HeLa cell clones should prove valuable hosts for various DT gene-containing vectors that express the toxin intracellularly . Update on aspirin desensitization for chronic rhinosinusitis with polyps in aspirin-exacerbated respiratory disease ( AERD ) . DB00945 -exacerbated respiratory disease ( AERD ) is a clinical condition which results in adverse upper and lower respiratory symptoms , particularly rhinitis , conjunctivitis , bronchospasm , and/or laryngospasm , following exposure to cyclooxygenase-1 ( P23219 ) inhibiting drugs , namely aspirin or nonsteroidal anti-inflammatory drugs ( NSAIDs ) . A provocative aspirin challenge is the gold standard for diagnosis of AERD . DB00945 desensitization and continuous aspirin therapy has been highly efficacious in those patients with suboptimal control of their disease on current available pharmacotherapy or those with other underlying conditions ( i.e. , cardiovascular disease ) who may require frequent treatment with aspirin or NSAIDs . This review article focuses on aspirin desensitization and the management of patients with AERD with a particular emphasis on outcomes in those patients with chronic rhinosinusitis and nasal polyposis . 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 . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . DB00435 modulates the frog heart ventricle morphodynamics . The aim of this work was to investigate in the avascular heart of the frog Rana esculenta the influence of nitric oxide ( NO ) on ventricular systolic and diastolic functions by using a novel image analysis technique . The external volume variations of the whole ventricle were monitored during the heart cycle by video acquisition ( visible light ) and analysed by an appropriately developed software with a specific formula for irregular convex solids . The system , which measures the rate of volume changes and the ejection fraction , directly determined the volumetric behaviour of the working frog heart after stimulation or inhibition of NOS-NOcGMP pathway . End-diastolic volume ( EDVext ) , end-systolic volume ( ESVext ) , contraction and relaxation velocities ( dV/dtsys and dV/dtdia , respectively ) , stroke volume ( SV ) and ejection fraction ( EF ) , were measured before and after perfusion with NOS substrate ( L-arginine ) , NO donor ( SIN-1 ) , cGMP analogue (8-Br-cGMP),NOS inhibitors ( NG-monomethyl-L-arginine , L-NMMA ; L-N(5)-(1-iminoethyl)-ornithine , L-NIO ; DB02207 ,7-NI ) and guanylyl cyclase inhibitor ( ODQ ) . The results showed that NO reduces ventricular systolicfunction improving diastolic filling , while NOS inhibition increases contractility impairing ventricular filling capacity . The presence of activated P29474 ( p- P29474 ) was morphologically documented , further supporting that the mechanical activity of the ventricular pump in frog is influenced by a tonic release of NOS-generated NO . Repair of benzo(a)pyrene diol epoxide- and UV-induced DNA damage in dihydrofolate reductase and adenine phosphoribosyltransferase genes of CHO cells . Using Uvr proteins we have quantified benzo(a)pyrene diol epoxide ( BPDE ) -DNA adduct formation and repair at the dihydrofolate reductase ( P00374 ) and adenine phosphoribosyltransferase ( P07741 ) genes in two Chinese hamster ovary cell lines : B-11 cells , which are 50-fold amplified for P00374 , and P01008 -2 cells , which are diploid for P00374 . We have found that : 1 ) BPDE-DNA adduct formation in different regions of the P00374 gene is proportional to the concentration of BPDE . 2 ) There is no significant difference in the repair of BPDE-DNA adducts between the coding and noncoding regions in either amplified or nonamplified P00374 gene domains . 3 ) Repair in the nonamplified P00374 gene is more efficient ( 30-40 % ) than in the amplified P00374 genes . 4 ) There are no significant differences of repair in the transcribed or nontranscribed strands of the P00374 gene . 5 ) BPDE-DNA adduct formation and repair in the P07741 gene in B-11 and P01008 -2 cells are the same . These results contrast those for the repair of cyclobutane pyrimidine dimers , which occurs preferentially in the transcribed strand of the P00374 gene and in which gene amplification appears to play no role . Identification of antithrombin-modulating genes . Role of O95461 , a gene encoding a bifunctional glycosyltransferase , in the secretion of proteins ? The haemostatic relevance of antithrombin together with the low genetic variability of P01008 , and the high heritability of plasma levels encourage the search for modulating genes . We used a hypothesis-free approach to identify these genes , evaluating associations between plasma antithrombin and 307,984 polymorphisms in the GAIT study ( 352 individuals from 21 Spanish families ) . Despite no SNP reaching the genome wide significance threshold , we verified milder positive associations in 307 blood donors from a different cohort . This validation study suggested O95461 , a gene encoding a protein with xylosyltransferase and glucuronyltransferase activities that forms heparin-like linear polysaccharides , as a potential modulator of antithrombin based on the significant association of one SNPs , rs762057 , with anti-FXa activity , particularly after adjustment for age , sex and P01008 rs2227589 genotype , all factors influencing antithrombin levels ( p = 0.02 ) . Additional results sustained this association . O95461 silencing inHepG2 and P29320 -EBNA cells did not affect P01008 mRNA levels but significantly reduced the secretion of antithrombin with moderate intracellular retention . Milder effects were observed on α1-antitrypsin , prothrombin and transferrin . Our study suggests O95461 as the first known modifier of plasma antithrombin , and proposes a new role for O95461 in modulating extracellular secretion of certain glycoproteins . A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 and P35354 ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . P13639 diphthamide is critical for translation of two IRES-dependent protein targets , P98170 and P09038 , under oxidative stress conditions . Elongation factor-2 ( eEF2 ) catalyzes the movement of the ribosome along the mRNA . A single histidine residue in eEF2 ( H715 ) is modified to form diphthamide . A role for eEF2 in the cellular stress response is highlighted by the fact that eEF2 is sensitive to oxidative stress and that it must be active to drive the synthesis of proteins that help cells to mitigate the adverse effects of oxidative stress . Many of these proteins are encoded by mRNAs containing a sequence called an " internal ribosomal entry site " ( IRES ) . Under high oxidative stress conditions diphthamide-deficient cells were significantly more sensitive to cell death . These results suggest that diphthamide may play a role in protection against the degradation of eEF2 . This protection is especially important in those situations in which eEF2 is necessary for the reprogramming of translation from global to IRES synthesis . Indeed , we found that the expression of X-linked inhibitor of apoptosis ( P98170 ) and fibroblast growth factor 2 ( P09038 ) , two proteins synthesized from mRNAs with IRESs that promote cell survival , is deregulated in diphthamide-deficient cells . Our findings therefore suggest that eEF2 diphthamide controls the selective translation of IRES-dependent protein targets P98170 and P09038 , critical for cell survival under conditions of oxidative stress . 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 . Cloning and analysis of a cDNA coding for bovine prothrombin . Poly(A)-RNA enriched for prothrombin was isolated by specific immunoprecipitation of bovine liver polysomes . P00734 consisted of about 8 % of the cell-free translation products of this RNA . A double-stranded cDNA was synthesized by using reverse transcriptase ( RNA-dependent DNA nucleotidyltransferase ) and made blunt-ended with nuclease S1 . After tailing with dCTP and terminal transferase , the double-stranded cDNA was annealed to pBR322 DNA that had been cleaved previously at the single Pst I site and similarly tailed with dGTP . The resulting plasmids were used to transform Escherichia coli strain P23921 under P09131 -EK1 conditions . Sixty-three tetracycline-resistant clones were obtained that hybridized to 32P-labeled cDNA synthesized from prothrombin-enriched mRNA . Recombinants containing cDNA to prothrombin mRNA sequences were screened by a solution hybridization assay with a [3H]cDNA synthesized from mRNA . This enriched mRNA was 50 % prothrombin mRNA , as determined by a reticulocyte lysate translation assay . Three positive clones were identified by this assay ; they contained bovine DNA inserts of 700 , 500 , and 400 base pairs . The DNA sequence of the 700-base-pair insert was then determined . This recombinant plasmid contained DNA coding for the carboxyl-terminal 160 residues of bovine prothrombin followed by a noncoding region of 119 base pairs and a poly(A) tail of 60 base pairs .

MH_train_1436

MH_train_1436

MH_train_1436

interacts_with DB00762?

multiple_choice

Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 receptor ( IL13Rα2 ) that differs from the physiological P24394 /IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL-13-PE ) encoding a mutated human P35225 fused to Pseudomonas exotoxin ( mhIL-13-PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 that binds to the physiological P24394 /IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL-13-PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL-13-PE and two protein formulations , hIL-13-PE used in clinical trials ( DB05305 ) and a second-generation mhIL-13-PE . DB05305 doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL-13-PE led to ∼40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL-13-PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 include its short half-life , which demanded frequent or continued administration , and binding to P24394 /IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM . Is there still room for large registrative trials in unselected cancer patients ? The case of anti-epidermal growth factor receptor antibodies in advanced non-small-cell lung cancer . DB09559 , a monoclonal antibody directed against P00533 , is currently under development as a treatment for advanced NSCLC . Two Phase III randomized trials are ongoing , testing the addition of necitumumab to first-line platinum-based chemotherapy . In the same setting , cetuximab produced a statistically significant but clinically modest benefit in the whole study population , and no solid data have been produced about predictive factors of efficacy . Will the difference in structure between the two antibodies be enough to obtain a clinically relevant advantage , making real progress in the treatment of advanced NSCLC ? Large Phase III trials in unselected patients risk demonstrating statistically significant results with debatable clinical relevance in the whole population , and the study of predictive factors is often left to subgroup analysis performed after the conduction of the trial . We do not need further ' me-too ' drugs , or drugs that produce a small benefit in the unselected population . On the contrary , the oncologic community needs drugs to be used with a proper selection of patients , to obtain larger , relevant benefits in molecularly characterized subgroups . Final results of randomized trials with necitumumab in advanced NSCLC are expected in a couple of years . Differential reconstitution of T cell subsets following immunodepleting treatment with alemtuzumab ( anti- P31358 monoclonal antibody ) in patients with relapsing-remitting multiple sclerosis . DB00087 ( anti- P31358 mAb ) provides long-lasting disease activity suppression in relapsing-remitting multiple sclerosis ( RRMS ) . The objective of this study was to characterize the immunological reconstitution of T cell subsets and its contribution to the prolonged RRMS suppression following alemtuzumab-induced lymphocyte depletion . The study was performed on blood samples from RRMS patients enrolled in the CARE-MS II clinical trial , which was recently completed and led to the submission of alemtuzumab for U.S . Food and Drug Administration approval as a treatment for RRMS . DB00087 -treated patients exhibited a nearly complete depletion of circulating P01730 (+) lymphocytes at day 7 . During the immunological reconstitution , P01730 (+)CD25(+)CD127(low) regulatory T cells preferentially expanded within the P01730 (+) lymphocytes , reaching their peak expansion at month 1 . The increase in the percentage of TGF-β1- , P22301 - , and P05112 -producing P01730 (+) cells reached a maximum at month 3 , whereas a significant decrease in the percentages of Th1 and Th17 cells was detected at months 12 and 24 in comparison with the baseline . A gradual increase in serum P13232 and P05112 and a decrease in Q16552 , Q96PD4 , Q9HBE4 , Q9GZX6 , and IFN-γ levels were detected following treatment . In vitro studies have demonstrated that P13232 induced an expansion of P01730 (+)CD25(+)CD127(low) regulatory T cells and a decrease in the percentages of Th17 and Th1 cells . In conclusion , our results indicate that differential reconstitution of T cell subsets and selectively delayed P01730 (+) T cell repopulation following alemtuzumab-induced lymphopenia may contribute to its long-lasting suppression of disease activity . 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 . P16066 regulates self-renewal and pluripotency of embryonic stem cells . Self-renewal and pluripotency of embryonic stem ( ES ) cells are maintained by several signaling cascades and by expression of intrinsic factors , such as Oct4 , Nanog and Sox2 . The mechanism regulating these signaling cascades in ES cells is of great interest . Recently , we have demonstrated that natriuretic peptide receptor A ( P16066 ) , a specific receptor for atrial and brain natriuretic peptides ( P01160 and DB04899 , respectively ) , is expressed in pre-implantation embryos and in ES cells . Here , we examined whether P16066 is involved in the maintenance of ES cell pluripotency . RNA interference-mediated knockdown of P16066 resulted in phenotypic changes , indicative of differentiation , downregulation of pluripotency factors ( such as Oct4 , Nanog and Sox2 ) and upregulation of differentiation genes . P16066 knockdown also resulted in a marked downregulation of phosphorylated Akt . Furthermore , P16066 knockdown induced accumulation of ES cells in the P55008 phase of the cell cycle . Interestingly , we found that P01160 was expressed in self-renewing ES cells , whereas its level was reduced after ES cell differentiation . Treatment of ES cells with P01160 upregulated the expression of Oct4 , Nanog and phosphorylated Akt , and this upregulation depended on P16066 signaling , because it was completely reversed by pretreatment with either an P16066 antagonist or a cGMP-dependent protein kinase inhibitor . These findings provide a novel role for P16066 in the maintenance of self-renewal and pluripotency of ES cells . TATA-driven transcriptional initiation and regulation of the rat serotonin P08908 receptor gene . The transcriptional initiation and regulation of the rat serotonin P08908 receptor gene were characterized . By three types of analyses , a single brain-specific site of transcriptional initiation was localized to -967 bp upstream of the translation initiation codon that is utilized both in hippocampus and in the rat raphe RN46A cell line . This major site of transcriptional initiation was located 58 bp downstream from a consensus TATA element , suggesting TATA-driven transcription of the rat P08908 receptor . To identify the promoter activity of the receptor gene , progressive 5' deletions of the -2,719/-117-bp fragment of the P08908 promoter linked to luciferase gene were transfected into P08908 -negative ( pituitary GH4C1 , Q9BTT4 myoblast , and P13671 glioma ) and P08908 -positive ( septal SN-48 and raphe RN46A ) cell lines . Enhancer regions were identified within a fragment between nucleotides -426 and -117 that selectively enhanced transcription in P08908 -positive cells . A nonselective enhancer/promoter that mediated expression in all cell lines was located upstream between -1,519 and -426 bp in a DNA segment containing consensus TATA , CCAAT , SP-1 , and AP-1 elements as well as a poly-GT26 dinucleotide repeat . Strong repression of transcription in all cell lines was conferred by the region upstream of -1,519 bp that contains a 152-bp DNA segment with > 80 % identity to RANTES , tumor necrosis factor-beta , and other immune system genes . Our results indicate that TATA-driven expression of the P08908 receptor is regulated by a novel proximal tissue-specific enhancer region , a nonselective promoter , and an upstream repressor region that is distinct from previously identified neuron-specific repressors . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . Determination of ancestral allele for possible human cancer-associated polymorphisms . To determine ancestral allele in possible cancer-associated polymorphisms , DNA samples from 10 chimpanzees ( Pan troglodytes ) were sequenced for alleles corresponding to 17 polymorphisms : 8 short tandem repeats [ P18510 ( alias IL-1RA ) variable number tandem repeat ( VNTR ) ; P04818 ( previously TS ) VNTR ; AR CAG repeat ; dinucleotide repeats of P22309 , IGF1 , P01579 ( alias P01579 ) , P03372 ( alias P03372 ) , and P00533 ] and 9 single nucleotide polymorphisms ( P03956 -1607 1G/2G , P08254 -1171 5A/6A , O15527 Ser326Cys , P05091 Gly487Lys , P04637 Arg72Pro , Q9UNQ0 Gln141Lys , P16455 Leu84Phe , P04179 Ala-9Val , and P42898 Ala222Val ) . No chimpanzee polymorphism corresponded to human P18510 VNTR ; the ancestral allele was a repeat lost in humans . Dinucleotide repeat polymorphisms of IGF1 , P01579 , P03372 , and P00533 were shared by chimpanzees , but the length of repeats tended to be longer in humans than in chimpanzees . This tendency was particularly evident for IGF1 . All of the SNPs tested are human-specific nucleotide changes . The ancestral allele 7A was shown to be lost in P08254 -1171 5A/6A . Thus , all of the possible cancer-associated polymorphisms tested have human-specific alleles , and the ancestral allele is lost in three polymorphisms ( P18510 VNTR , P22309 CA repeat , and P08254 -1171 5A/6A ) , suggesting a possible involvement of human-specific alleles in cancer susceptibility . 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 . The stop transfer sequence of the human UDP-glucuronosyltransferase 1A determines localization to the endoplasmic reticulum by both static retention and retrieval mechanisms . Human UDP-glucuronosyltransferase 1A ( P22309 ) isoforms are endoplasmic reticulum ( ER ) -resident type I membrane proteins responsible for the detoxification of a broad range of toxic phenolic compounds . These proteins contain a C-terminal stop transfer sequence with a transmembrane domain ( TMD ) , which anchors the protein into the membrane , followed by a short cytosolic tail ( CT ) . Here , we investigated the mechanism of ER residency of P22309 mediated by the stop transfer sequence by analysing the subcellular localization and sensitivity to endoglycosidases of chimeric proteins formed by fusion of P22309 stop transfer sequence ( TMD/CT ) with the ectodomain of the plasma membrane P01730 reporter protein . We showed that the stop transfer sequence , when attached to C-terminus of the P01730 ectodomain was able to prevent it from being transported to the cell surface . The protein was retained in the ER indicating that this sequence functions as an ER localization signal . Furthermore , we demonstrated that ER localization conferred by the stop transfer sequence was mediated in part by the KSKTH retrieval signal located on the CT . Interestingly , our data indicated that P22309 TMD alone was sufficient to retain the protein in ER without recycling from Golgi compartment , and brought evidence that organelle localization conferred by P22309 TMD was determined by the length of its hydrophobic core . We conclude that both retrieval mechanism and static retention mediated by the stop transfer sequence contribute to ER residency of P22309 proteins . 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 . [ 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 . DB08889 can induce tumor cell death through selective inhibition of the chymotrypsin-like activity of the proteasome . DB08889 is a proteasome inhibitor in clinical development that primarily targets the chymotrypsin-like ( CT-L ) subunits in both the constitutive proteasome ( c20S ) and the immunoproteasome ( i20S ) . To investigate the impact of inhibiting the CT-L activity with carfilzomib , we set out to quantitate the levels of CT-L subunits beta5 from the c20S and P28062 from the i20S in normal and malignant hematopoietic cells . We found that the i20S is a major form of the proteasome expressed in cells of hematopoietic origin , including multiple myeloma ( MM ) CD138+ tumor cells . Although specific inhibition of either P28062 or beta5 alone was insufficient to produce an antitumor response , inhibition of all proteasome subunits was cytotoxic to both hematologic tumor cells and peripheral blood mononuclear cells . However , selective inhibition of both beta5 and P28062 was sufficient to induce an antitumor effect in MM , non-Hodgkin lymphoma , and leukemia cells while minimizing the toxicity toward nontransformed cells . In MM tumor cells , CT-L inhibition alone was sufficient to induce proapoptotic sequelae , including proteasome substrate accumulation , Noxa and caspase 3/7 induction , and phospho-eIF2alpha suppression . These data support a hypothesis that hematologic tumor cells are uniquely sensitive to CT-L inhibition and provide a mechanistic understanding of the clinical safety profile and antitumor activity of proteasome inhibitors . Inhibition of rat liver UDP-glucuronosyltransferase by silymarin and the metabolite silibinin-glucuronide . The inhibitory effects of silymarin , its main constituent silibinin and the metabolite silibinin-glucuronide on UDP-glucuronosiltransferase ( P78381 ) were evaluated in rat hepatic microsomes . Three substrates were chosen to cover both P22309 and UGT2B family isozymes : bilirubin ( substrate of P22309 ) , p-nitrophenol ( P19224 ) and ethinylestradiol ( UGT2B1 and 2B3 for position C17 and P22309 for position P01024 ) . The study of p-nitrophenol and bilirubin glucuronidation indicated that silymarin ( SM ) and silibinin glucuronide ( SB-G ) were enzyme inhibitors . The kinetic analysis showed that the type of inhibition was competitive in all cases and the Ki obtained were : for p-nitrophenol glucuronidation , KiSB-Gapp : 14+/-1 microg/ml and KiSMapp : 51+/-10 microg/ml and for bilirubin glucuronidation , KiSB-Gapp : 16+/-3 microg/ml . In turn , ethinylestradiol glucuronidation was not affected by any of the compounds studied suggesting that the inhibitory effect was restricted to P22309 isozymes . Similar studies performed using human hepatic microsomes showed that SM and SB-G were also inhibitors of human P22309 isozymes . In conclusion , administration of silymarin or its main constituent silibinin could lead to the decrease in the glucuronidation of substrates whose conjugation depends on P22309 isozymes in a process mediated by silibinin-glucuronide , though their effect in humans needs further investigation . Second-generation epidermal growth factor receptor tyrosine kinase inhibitors in lung cancers . P00533 mutations identify patients who are more likely to respond to treatment with epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors ( TKIs ) than cytotoxic chemotherapy . The distinct success of the first-generation P00533 TKIs erlotinib and gefitinib has been accompanied by the observation that acquired resistance to these treatments develops after a median of 1 year of treatment . Newer , second-generation P00533 TKIs have been developed with the intent to delay or overcome acquired resistance by the broader inhibition of kinases ( eg , P04626 and vascular endothelial growth factor receptor ) and/or altering the interactions with P00533 through irreversibly binding to the kinase domain . This article discusses many of these agents ( including afatinib , dacomitinib , DB05007 , AP26113 , and CO-1686 ) which have the potential for greater efficacy compared with first-generation P00533 TKIs , and may also have clinical activity against other oncogenic mutations within the P00533 family , including P04626 . DB00075 -induced anergy in cloned human Th0 , Th1 , and Th2 cells . In the mouse , activation of T cells by T cell receptor ( TCR ) crosslinking with anti-CD3 antibodies in the absence of a costimulatory signal induces Th1 but not Th2 cell anergy . Furthermore , anti-CD3 induces anergy of Th1- but not Th2-type lymphokine secretion in Th0 cells . This study was designed to determine whether this is also the case in man . Human rye grass allergen Lol p I-specific cloned P01730 + T helper cells of subtypes Th0 , Th1 , and Th2 were treated with immobilized anti-CD3 . The cells were rested for 4 days and then activated under optimal conditions with antigen and antigen-presenting cells ( APCs ) . Cell proliferation and P60568 , P01579 , and P05112 secretion was determined to test for the anergic state . The initial anti-CD3 treatment induced cell proliferation , P60568 , P01579 , and/or P05112 secretion by T cells of all three subsets which was followed by an anergic state in Th0 , Th1 , and Th2 cells as shown by a 51 to > 94 % decrease in cell proliferation and P01579 and/or P05112 secretion after subsequent P25054 and Lol p I activation . Addition of P60568 or P05112 during anti-CD3 treatment of the cells did not prevent unresponsiveness . However , the addition of P60568 but not P05112 during P25054 and Lol p I stimulation partially reversed the anergic state . These data demonstrate that , contrary to the mouse , cloned T cells of all three human T helper cell subtypes are anergized by anti-CD3 TCR activation in the absence of costimulatory signals . The fact that human Th2 cells can be anergized may be important for the development of new treatments in Th2-mediated allergic disorders . Preimplantation genetic diagnosis for cancer predisposition syndromes . OBJECTIVES : Mutations in the P25054 , P35240 and P38398 genes cause adult-onset cancer predisposition syndromes . Prenatal diagnosis ( P01160 ) and selective pregnancy termination for adult-onset disorders is emotionally difficult and , in some cases , socially not well accepted . Preimplantation genetic diagnosis ( P52209 ) appears as an attractive alternative to P01160 , as it ensures the establishment of a pregnancy free of the mutation from the onset , circumventing the potentially difficult decision of termination of pregnancy . METHODS : Development of single-cell PCRs using Epstein-Barr virus transformed lymphoblasts as single-cell model , followed by clinical application in P52209 . RESULTS : A total of five duplex-PCRs were developed , three for adenomatous polyposis of the colon ( P25054 ) , one for neurofibromatosis type 2 ( P35240 ) and one for inherited breast and ovarian cancer caused by P38398 mutations . Eleven clinical cycles were performed , resulting in the birth of an unaffected girl . For one of the couples undergoing P52209 for P35240 , a spontaneous pregnancy ensued after five unsuccessful P52209 cycles . The couple underwent chorionic villus sampling ( CVS ) and the application of the same protocol as used during P52209 showed an unaffected fetus . CONCLUSION : In this work , we present the development and clinical application of P52209 for three cancer predisposition syndromes . Bipartite regulation of different components of the MHC class I antigen-processing machinery during dendritic cell maturation . Dendritic cells ( DC ) are professional antigen-presenting cells ( P25054 ) which proceed from immature to a mature stage during their final differentiation . Immature DC are highly effective in terms of antigen uptake and processing , whereas mature DC become potent immunostimulatory cells . Until now , the expression profiles of the major components of the MHC class I antigen-processing machinery ( APM ) during DC development have not been well characterized . In this study , the mRNA and protein expression levels of the P01579 inducible proteasome subunits , of the proteasome activators PA28 , and of key components required for peptide transport and MHC class I-peptide complex assembly have been evaluated in immature and mature stages of human monocyte-derived DC using semiquantitative RT-PCR and Western blot analyses . The P01579 -responsive immunoproteasome subunits P28065 , P28062 and P40306 are up-regulated in immature DC , whereas the other components of the MHC class I presentation machinery , such as PA28 , TAP , tapasin , and HLA heavy and light chains , were found to be more abundant in mature DC . These findings support the hypothesis that immature DC produced by the differentiation of monocytes in response to P05112 and granulocyte macrophage colony stimulating factor first increase their capacity to capture antigens and process them into peptides , thereby switching from housekeeping to immunoproteasomes , while mature DC rather up-regulate the components required for peptide translocation and MHC class I-peptide complex formation , and thus specialize in antigen presentation . Our results establish that MHC class I , like MHC class II surface expression , is markedly regulated during DC development and maturation .

MH_train_1437

MH_train_1437

MH_train_1437

interacts_with DB00741?

multiple_choice

DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . 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 . IL-1 does not reverse the anti-proliferative effect of aspirin in breast cancer cells . OBJECTIVES : Interleukin-1 ( IL-1 ) is a multifunctional proinflammatory cytokine . There have been studies suggesting a role in affecting growth and invasiveness of malignant breast cells by either blocking or stimulating growth of cultured MCF-7 breast cancer cells . This effect may be mediated by induction of P35354 . DB00945 is an inhibitor of P35354 and has been implicated , with other non-steroidal anti-inflammatory drugs ( NSAIDS ) in prevention and treatment of breast cancer . In this study the in vitro effects of IL-1 and aspirin on growth of MCF-7 human breast cancer cells was examined . METHODS : MCF-7 cells were treated with various concentrations of IL-1 and aspirin alone and in combination . Cell growth was assessed by cell number measurement . RESULTS : DB00945 significantly decreased growth rate in a dose-dependent manner , alone and as a combined treatment with IL-1 with a maximum reduction in growth rate at 300 mg/ml ( P < 0.05 ) . Treatment with IL-1 alone showed no significant effect on growth rate of MCF-7 cells ( P > 0.05 ) . CONCLUSION : This study confirms that aspirin suppresses the proliferation rate of MCF-7 cells both as a single agent and in combination with IL-1 . It also suggests that IL-1 alone does not stimulate or inhibit growth of MCF-7 cells . Induction of tumoricidal activity in mouse peritoneal macrophages by ginseng polysaccharide . This study examined the effects of ginseng polysaccharide ( GPS ) on mouse peritoneal macrophage ( PM ) -mediated cytotoxicity towards K562 , HL-60 , or KG1alpha cells . GPS had no direct effect on killing of tumor cells . However , when mouse PMs were treated with GPS , cytotoxic activity against K562 , HL-60 , or KG1alpha cells was significantly induced . In addition , phagocytic activity was enhanced in GPS-treated mouse PMs compared to the control . The expressions of CD(68) , O14561 and alpha-ANE in mouse PMs were increased by the treatment with GPS . Moreover , the levels of cytokines , including tumor necrosis factor-alpha ( P01375 ) , interleukin-1 ( IL-1 ) , P05231 were increased and the production of nitric oxide ( NO ) was enhanced . Taken together , these results suggest that GPS possess a potent antitumor activity by stimulating macrophage and a potentiality as an immunomodulator against diseases such as cancer . DB00160 aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis . Plant peroxisomal glyoxylate aminotransferases play central roles within the photorespiratory pathway . Genes encoding glyoxylate aminotransferases have been isolated from several animals and microbes , but only recently have plant homologs been identified . Three Arabidopsis homologs of alanine (Ala):glyoxylate aminotransferase 2 ( Q9BYV1 ) contain a putative type 1 peroxisomal targeting signal ( PTS1 ) , but the metabolic significance of these Q9BYV1 homologs is unknown . P19440 and P36268 are Ala aminotransferase ( AlaAT ) homologs from Arabidopsis that represent another type of glyoxylate aminotransferase . These proteins are class I aminotransferases , each containing a putative PTS1 . P19440 and P36268 are members of a small family of AlaATs in Arabidopsis . When expressed as recombinant proteins in Escherichia coli , P19440 and P36268 displayed biochemical characteristics very similar to one another , and to the Arabidopsis protein purified from leaves . Four aminotransferase activities were specifically associated with P19440 and P36268 , using the substrate pairs glutamate ( DB00142 ):glyoxylate , Ala:glyoxylate , DB00142 :pyruvate , and Ala:2-oxoglutarate . P19440 and P36268 may have partially redundant functions ; transcripts of both genes were detected in many of the same tissues . Although DB00142 :glyoxylate aminotransferase ( P19440 ) activity has been observed in several locations in different plants and algae , including the cytoplasm and mitochondria , our subcellular fractionation data indicate that P19440 activity was exclusively peroxisomal in Arabidopsis . Thus , glyoxylate aminotransferase reactions in plant peroxisomes appear to be catalyzed by at least two distinct types of aminotransferases : an AGT1 homolog with serine:glyoxylate aminotransferase activity ( A.H. Liepman , L.J. Olsen [ 2001 ] Plant J 25 : 487-498 ) , and a pair of closely related , potentially redundant AlaAT homologs with P19440 activity . 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 . 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 . Safety , pharmacokinetics and pharmacodynamics of four-hour intravenous infusions of eritoran in healthy Japanese and Caucasian men . DB04933 , a synthetic analogue of lipid A , has been shown to bind to O00206 /MD-2 complex and thereby block the interaction of endotoxins with O00206 . We report here the results of a study conducted to assess the single-dose safety and tolerability , as well as the pharmacokinetics and pharmacodynamics , of eritoran infusion in Japanese and Caucasian healthy adult men . Sixty-four men ( aged 20-45 years ; body mass index 18-30 kg/m(2) ) were randomized into four groups : 4-mg total dose ( six Japanese and six Caucasian men ) ; 12-mg total dose ( 12 Japanese and 12 Caucasian men ) ; 28-mg total dose ( six Japanese and six Caucasian men ) ; and placebo ( eight Japanese and eight Caucasian men ) . DB04933 in single doses up to 28 mg over 4 h was well tolerated , with no apparent ethnic differences noted . Plasma concentrations were slightly higher in Japanese versus Caucasian men ; these differences were not significant after adjustment for differences in body mass ( clearance : approximately 1.2 ml/h/kg ; volume of distribution at steady state : approximately 0.07 l/kg ) . The ex vivo endotoxin inhibitory activity of eritoran was similar in Japanese and Caucasian men . The data do not indicate any need for clinical dose adjustment for possible ethnic-based differences in drug distribution or metabolism . Immunomodulating effects of cefodizime on Klebsiella pneumoniae-stimulated neutrophils . To explore the immunoregulating effects of cefodizime on neutrophils and its mechanisms , we detected the expression of some cytokines secreted by neutrophils after heat-killed Klebsiella pneumoniae induced inflammation . We analyzed the changes of signal transduction in this process by detecting the mRNA expression of toll like receptor 4 ( O00206 ) and the inhibitor factor of kappaBalpha ( I-kappaBalpha ) expressed by neutrophils . The activity of nuclear factor kappaB ( NF-kappaB ) in neutrophils was also assayed by electrophoretic mobility shift assay ( EMSA ) . We found cefodizime increased neutrophil production of P01375 , IL-1beta in the early stage of inflammation , which was in agreement with the enhanced mRNA expression of O00206 and DNA-binding activity of NF-kappaB . Taken together , the immunomodulating effects of cefodizime on cytokine production of K. pneumoniae stimulated neutrophils is possibly due to its regulation of O00206 mRNA expression and DNA binding activities of NF-kappaB through the LPS- O00206 -NF-kappaB pathway . 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 . Antiinflammatory effect of lactic acid bacteria : inhibition of cyclooxygenase-2 by suppressing nuclear factor-kappaB in Raw264.7 macrophage cells . Lactobacillus casei 3260 ( L. casei 3260 ) was evaluated in relation to the inflammatory response mediated by lipopolysaccharide ( LPS ) -induced nuclear factor-kappaB ( NF-kappaB ) and cyclooxygenase-2 ( P35354 ) expression in Raw264.7 macrophage cells . The treatment of Raw264.7 cells with L. casei 3260 significantly inhibited the secretion of tumor necrosis factor-alpha ( P01375 ) and prostaglandins E2 ( DB00917 ) , followed by suppression of P35354 . To clarify the molecular mechanism , the inhibitory effect of L. casei 3260 on the NF-kappaB signaling pathway was examined based on the luciferase reporter activity . Although the treatment of Raw264.7 cells with L. casei 3260 did not affect the transcriptional activity of NF-kappaB , it did inhibit NF-kappaB activation , as determined by the cytosolic p65 release and degradation of I-kappaBalpha . Therefore , these findings suggest that the suppression of P35354 through inhibiting the NF-kappaB activation by LPS may be associated with the antiinflammatory effects of L. casei 3260 on Raw264.7 cells . Beta-ketoacyl acyl carrier protein reductase ( FabG ) activity of the fatty acid biosynthetic pathway is a determining factor of 3-oxo-homoserine lactone acyl chain lengths . The two acyl-homoserine lactones ( AHLs ) N-(butyryl)-L-homoserine lactone and N-[3-oxododecanoyl]-L-homoserine lactone ( 3-oxo-C(12)- Q05469 ) are required for quorum sensing in Pseudomonas aeruginosa . These AHLs derive their invariant lactone rings from S-adenosylmethionine and their variable acyl chains from the cellular acyl-acyl carrier protein ( O14561 ) pool . This reaction is catalysed by specific AHL synthases , which exhibit acyl chain specificity . Culture supernatants of P. aeruginosa contain multiple 3-oxo-AHLs that differ in their acyl chain lengths but their physiological role , if any , remains unknown . An in vitro fatty acid-3-oxo-AHL synthesis system was established utilizing purified P. aeruginosa Fab proteins , O14561 and the LasI 3-oxo-AHL synthase . In the presence of excess protein , substrates and cofactors , this system produced almost exclusively 3-oxo-C(12)- Q05469 . When the beta-ketoacyl- O14561 reductase ( FabG ) catalysed step was made rate-limiting by switching from the preferred NADPH cofactor to DB00157 , increased levels of short chain 3-oxo-AHLs were produced , presumably because shorter-chain ketoacyl-ACPs accumulated and thus became LasI substrates . Consistent with these in vitro observations , a fabG(Ts) mutant produced increased amounts of 3-oxo-AHLs in vivo . Thus , in vitro and in vivo evidence indicated that modulation of FabG activity of the fatty acid biosynthetic pathway may determine the acyl chain lengths of these 3-oxo-AHLs and that the LasI 3-oxo-AHL synthase is sufficient for their synthesis . 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 . Lactobacillus plantarum TN8 exhibits protective effects on lipid , hepatic and renal profiles in obese rat . This study aimed to first investigate the immuno-modulatory effects of six newly isolated lactic acid bacteria ( Q9GZY6 ) on the peripheral blood mononuclear cells ( PBMC ) of Wistar rats . Except for Lactobacillus plantarum TN8 , all the other strains were noted to induce high levels of pro-inflammatory cytokine IL-12 and low levels of anti-inflammatory cytokine P22301 . The strains also generated low ratios of P22301 /IL-12 cytokine . Strain TN8 was , on the other hand , noted to induce an increase in anti-inflammatory P22301 cytokine secretion rates and a decrease in pro-inflammatory IL-12 , IFN-γ and P01375 -α cytokine production . The oral administration of TN8 improved the hepatic and urinary functions of obese rats by inducing decreases ( P < 0.05 ) in alanine amino transferase ( ALAT ) , gamma glutamyl transferase ( P19440 ) , plasmatic triglycerides , total cholesterol concentrations , creatinine , urea , and body weight when compared to the control group of animals that underwent an increase in aspartate amino transferase ( ASAT ) and high density lipoprotein ( HDL ) . Overall , the findings indicate that strain TN8 exhibited a number of attractive properties that might open new promising opportunities for the improvement of various parameters related to animal health performance and the avoidance of antibiotics and drugs as promoting factors . DB08439 . Pharmacia corp . Pharmacia ( formerly Monsanto ) is developing parecoxib , an injectable P35354 inhibitor , for the management of post-surgical acute pain [ 287279 ] , [ 313957 ] . By January 1999 , the compound was in phase III trials for this indication [ 312280 ] . In October 2000 , Pharmacia submitted an NDA for parecoxib sodium for the management of acute pain to the FDA . The company anticipated a 12-month review of the NDA [ 387654 ] but received a ' not approvable ' letter in July 2001 , indicating there were deficiencies in the filing ; at this time , Pharmacia anticipated refiling before the end of 2002 [ 415668 ] . Under a license agreement with Pharmacia Corp , parecoxib ( designated YM-177 ) is being developed in Japan by Yamanouchi [ 392030 ] . Prior to the FDA ruling , in June 2000 , the company anticipated that the compound would be launched by 2001 [ 370466 ] . In March 2000 , Merrill Lynch predicted that parecoxib would be filed in the third quarter of 2000 [ 361969 ] , [ 382577 ] . By May 2001 , the analysts revised their predictions to launch in 2002 [ 411811 ] . Estrogen receptors mediate rapid activation of phospholipase C pathway in the rat endometrium . The aim of the present study was to investigate the activation of rapid signaling events by 17β-estradiol in the rat uterus . 17β-Estradiol induced a rapid increase of total [ 3H ] -inositol phosphate accumulation in the whole uterus and endometrium , but not in the myometrium . The effect of 17β-estradiol in the endometrium was blocked by phospholipase C ( P98160 ) inhibitor ( U73122 ) , estrogen receptors antagonist ( DB00947 ) , exportin O14980 inhibitor ( leptomycin B ) and selective inhibitor of the P12931 family of protein tyrosine kinases ( Q99463 ) . Furthermore , a selective agonist of P03372 ( PPT ) and a selective agonist of Q99527 ( G-1 ) also induced a rapid increase of total [ (3)H ] -inositol phosphate accumulation in the endometrium . The G-1 effects were blocked by Q99527 antagonist ( G-15 ) . 17β-Estradiol and G-1 promoted an additive effect on total [ 3H ] -inositol phosphate accumulation . In conclusion , the present results indicate that a rapid activation of the P98160 -mediated phosphoinositide hydrolysis occurred in the rat endometrium after 17β-estradiol stimulation , and this effect was mediated by P03372 that underwent nuclear export after hormone stimulation , and that Q99527 activation may play an additive role for this response . These rapid actions might be one of the key steps that mediate the estrogen-dependent activation of cellular events in the endometrium . Lipopolysaccharide-induced NF-kappaB activation and cytokine release in human alveolar macrophages is PKC-independent and TK- and PC- P98160 -dependent . A critical feature of sepsis-induced adult respiratory distress syndrome ( ARDS ) is the release of cytokines ( such as interleukin [ IL ] -6 , P10145 , and tumor necrosis factor [ P01375 ] ) from endotoxin ( lipopolysaccharide [ LPS ] ) -activated alveolar macrophages ( AM ) . Nuclear factor kappa B ( NF-kappaB ) is activated in AM from patients with ARDS , and it is essential for the transcription of many cytokine genes . In these studies , we evaluated the regulation of LPS-induced cytokine release and the activation of NF-kappaB in human AM . We found that the activation of NF-kappaB and the release of P05231 , P10145 , and P01375 from AM exposed to LPS was protein kinase C-independent and tyrosine kinase- and phosphatidylcholine-specific phospholipase C-dependent . We also found that LPS-induced activation of NF-kappaB was enhanced in AM cultured in serum or in the presence of LPS-binding protein , simulating conditions in the lung that are present in ARDS . In addition , LPS triggered the activation of several different NF-kappaB complexes in AM , and different forms of NF-kappaB bound to the P05231 , P10145 , and P01375 promoter sequences . These observations suggest that physiologic abnormalities present in the lungs of patients with ARDS facilitate the activation of NF-kappaB and local release of cytokines .

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interacts_with DB00834?

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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 . Current perspectives on role of chromatin modifications and deacetylases in lung inflammation in P48444 . Chromatin modifications and epigenetic regulation are critical for sustained and abnormal inflammatory response seen in lungs of patients with chronic obstructive pulmonary disease ( P48444 ) because the activities of enzymes that regulate these epigenetic modifications are altered in response to cigarette smoke . Cigarette smoke induces chromatin modifications and epigenetic changes by causing post-translational modifications of histone acetyltransferases , and histone/non-histone deacetylases ( HDACs ) , such as Q92769 and sirtuin 1 ( Q96EB6 ) , which leads to chromatin remodeling . In this review , we discussed the current knowledge on cigarette smoke/oxidants-induced post-translational modifications of deacetylases ( Q92769 and Q96EB6 ) , disruption of Q92769 / Q96EB6 -RelA/p65 corepressor complex associated with acetylation of RelA/p65 , and chromatin modifications ( histone H3 phospho-acetylation ) leading to sustained pro-inflammatory gene transcription . Knowledge on molecular mechanisms of epigenetic changes in abnormal lung inflammation will help in understanding the pathophysiology of P48444 which may lead to the development of novel epigenetic therapies in the near future . Integration of endocannabinoid and leptin signaling in an appetite-related neural circuit . Recently developed therapeutics for obesity , targeted against cannabinoid receptors , result in decreased appetite and sustained weight loss . Prior studies have demonstrated P21554 receptors ( CB1Rs ) and leptin modulation of cannabinoid synthesis in hypothalamic neurons . Here , we show that depolarization of perifornical lateral hypothalamus ( LH ) neurons elicits a CB1R-mediated suppression of inhibition in local circuits thought to be involved in appetite and " natural reward. " The depolarization-induced decrease in inhibitory tone to LH neurons is blocked by leptin . Leptin inhibits voltage-gated calcium channels in LH neurons via the activation of janus kinase 2 ( O60674 ) and of mitogen-activated protein kinase ( MAPK ) . Leptin-deficient mice are characterized by both an increase in steady-state voltage-gated calcium currents in LH neurons and a CB1R-mediated depolarization-induced suppression of inhibition that is 6-fold longer than that in littermate controls . Our data provide direct electrophysiological support for the involvement of endocannabinoids and leptin as modulators of hypothalamic circuits underlying motivational aspects of feeding behavior . 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 . 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 . P37231 : a nuclear receptor with affinity for cannabinoids . An increasing number of cannabinoid actions are being reported that do not appear to be mediated by either P21554 or CB2 , the known cannabinoid receptors . One such example is the synthetic analog ajulemic acid ( AJA ) , which shows potent analgesic and anti-inflammatory effects in rodents and humans . AJA binds weakly to P21554 only at concentrations many fold higher than its therapeutic range , and is , therefore , completely free of psychotropic effects in both normal subjects and pain patients suggesting the involvement of a target site other than P21554 . AJA as well as several other cannabinoids appear to have profound effects on cellular lipid metabolism as evidenced by their ability to transform fibroblasts into adipocytes where the accumulation of lipid droplets can be readily observed . Such transformations can be mediated by the activation of the nuclear receptor P37231 . A variety of small molecule ligands including AJA have been shown to induce the activation of P37231 and , in some cases this has led to the introduction of clinically useful agents . It is suggested that P37231 may serve a receptor function for certain actions of some cannabinoids . 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 . 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 . 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 . [ Rapid inhibitory effect of glucocorticoids on peak of [Ca2+]i and P98160 in airway smooth muscle ] . OBJECTIVE : In this study , we pretreated the mice ASMCs by dexamethasone ( DB00514 ) within 10 min , to test the peak of [Ca2+]i and phospho-PLCbeta ( ser1105 ) in the cells by treated with Ach . METHODS : The peak of [Ca2+]i was measured by Fura-2/AM methods and the phospho-PLCbeta-ser1105 was by Western blot , and compared with dexamethasone pretreated groups . P04150 antagonist DB00834 and the protein synthesis inhibitor cycloheximide groups were settled in our study . RESULTS : Glucocorticoids ( GCs ) significantly decreased the resting values and peak of [Ca2+]i elevation and elevated the intracellular levels of phospho-PLCbeta ( ser1105 ) in 10 min . Neither the DB00834 nor cycloheximide could alter the inhibitory effects of glucocorticoids stated above . CONCLUSION : Our results demonstrate that glucocorticoids exert rapid inhibitory effects . The series of signal changes in this process that restrain the peak of [Ca2+]i may be responsible for the rapid nongenomic inhibitory effects of GCs by reducing the activity of P98160 . Anandamide regulates neuropeptide release from capsaicin-sensitive primary sensory neurons by activating both the cannabinoid 1 receptor and the vanilloid receptor 1 in vitro . The effect of anandamide , which activates both the cannabinoid 1 ( P21554 ) receptor and the vanilloid receptor 1 ( Q8NER1 ) , was studied on calcitonin gene-related peptide ( P80511 ) release from cultured primary sensory neurons , the majority of which coexpress the P21554 receptor and Q8NER1 . Concentrations of anandamide < 1 micro m produced a small but significant P21554 receptor-mediated inhibition of basal P80511 release while higher concentrations induced Q8NER1 -mediated P80511 release . The excitatory effect of anandamide was potentiated by the P21554 receptor antagonist SR141716A . In the presence of SR141716A at concentrations < 100 nm , anandamide was equipotent with capsaicin in stimulating P80511 release . However , at higher concentrations anandamide produced more P80511 release than equimolar concentrations of capsaicin . Three and ten nanomolar anandamide inhibited the capsaicin-evoked P80511 release . In the presence of SR141716A , treatments which activated protein kinase A , protein kinase C and phospholipase C significantly potentiated the anandamide-evoked P80511 release at all anandamide concentrations . Although this potentiation was reduced when the P21554 receptor antagonist was omitted from the buffer , the P80511 release evoked by 300 nm and 1 micro m anandamide was still significantly larger than that seen with nonpotentiated cells . These data indicate that anandamide may regulate P80511 release from capsaicin-sensitive primary sensory neurons in vivo , and that the net effect of anandamide on transmitter release from capsaicin-sensitive primary sensory neurons depends on the concentration of anandamide and the state of the P21554 receptor and Q8NER1 . These findings also suggest that anandamide could be one of the molecules responsible for the development of inflammatory heat hyperalgesia . DB03886 -dependent hyperphenylalaninemia due to deficiency of 6-pyruvoyl tetrahydropterin synthase . We describe the clinical , neurologic , and biochemical findings in 10 patients with 6-pyruvoyl tetrahydropterin synthase ( 6- Q03393 ) deficiency from seven families , all of whom originate from one large tribe in Saudi Arabia . This deficiency presents with severe , early onset of failure to thrive , neurologic deterioration , and morbidity and mortality secondary to repeated episodes of bronchopneumonia or cardiorespiratory abnormalities . The urinary pterin excretion pattern indicates deficient activity of 6- Q03393 , which has been confirmed by direct enzyme assay in red blood cells of three patients . We treated our patients with combined use of tetrahydrobiopterin 20 mg/kg/d , L-dihydroxyphenylalanine 15 mg/kg/d , carbidopa 3.75 mg/kg/d , and L-5-hydroxytryptophan 5 mg/kg/d . Neurologic findings improved significantly in all after 5 to 24 months . Although head circumference and weight returned to the lower limit of normal in four , height normalized only in one of seven patients . Despite an unrestricted diet during combined therapy , blood phenylalanine and urinary excretion of neopterin and biopterin returned to normal . 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 . The role of peroxisome proliferator-activated receptor-gamma and estrogen receptors in genistein-induced regulation of vascular tone in female rat aortas . BACKGROUND : DB01645 ( Q86UG4 ) is a phytoestrogen that binds estrogen receptors ( ER ) to produce a protective cardiovascular effect . It also has been shown binding peroxisome proliferator-activated receptor-gamma ( P37231 ) . METHODS : In the present study , we assessed the role of P37231 and ER in Q86UG4 -mediated regulation of vascular tone in female rat aortas by in vitro tension measurement , immunohistochemistry , immunofluorescence , immunoprecipitation and Western blot analysis . RESULTS : In aortas pretreated with GW9662 ( inhibitor of P37231 ) , ICI182780 ( inhibitor of ER ) and a combination of GW9662 and ICI182780 , the magnitudes of Q86UG4 -induced dilatation were attenuated . N(G)-nitro-L-arginine methyl ester had a similar effect . Q92731 and P37231 colocalized in all 3 layers of the aortas , while P03372 and P37231 colocalized only in the vascular endothelium and adventitia . In Q86UG4 -treated whole-cell protein samples , we demonstrated coimmunoprecipitation of Q92731 ( not P03372 ) and P37231 . The expression of Q92731 and P37231 in nuclear protein from Q86UG4 -treated samples increased , which was partially reversed by either GW9662 or ICI182780 and more efficiently reversed using a combination of GW9662 and ICI182780 . CONCLUSION : Our findings suggest that Q86UG4 can relax phenylephrine-induced vascular contraction in female rat aortas , which is mediated by P37231 and Q92731 . P04150 and histone deacetylase-2 mediate dexamethasone-induced repression of P98088 gene expression . Airway occlusion in obstructive airway diseases is caused in part by the overproduction of secretory mucin glycoproteins through the up-regulation of mucin ( MUC ) genes by inflammatory mediators . Some pharmacological agents , including the glucocorticoid dexamethasone ( DB00514 ) , repress mucin concentrations in lung epithelial cancer cells . Here , we show that DB00514 reduces the expression of P98088 , a major airway mucin gene , in primary differentiated normal human bronchial epithelial ( NHBE ) cells in a dose-dependent and time-dependent manner , and that the DB00514 -induced repression is mediated by the glucocorticoid receptor ( GR ) and two glucocorticoid response elements ( GREs ) in the P98088 promoter . The pre-exposure of cells to DB00834 , a GR antagonist , and mutations in either the GRE3 or GRE5 cis-sites abolished the DB00514 -induced repression . Chromatin immunoprecipitation ( ChIP ) assays showed a rapid temporal recruitment of GR to the GRE3 and GRE5 cis-elements in the P98088 promoter in NHBE and in A549 cells . Immunofluorescence showed nuclear colocalization of GR and histone deacetylase-2 ( Q92769 ) in P98088 -expressing NHBE cells . ChIP also showed a rapid temporal recruitment of Q92769 to the GRE3 and GRE5 cis-elements in the P98088 promoter in both cell types . The knockdown of Q92769 by Q92769 -specific short interfering RNA prevented the DB00514 -induced repression of P98088 in NHBE and A549 cells . These data demonstrate that GR and Q92769 are recruited to the GRE3 and GRE5 cis-sites in the P98088 promoter and mediate the DB00514 -induced cis repression of P98088 gene expression . A better understanding of the mechanisms whereby glucocorticoids repress P98088 gene expression may be useful in formulating therapeutic interventions in chronic lung diseases . DB00092 therapy reduces the effector T-cell population in lesional psoriatic epidermis . DB00092 , a LFA-3/IgG1 fusion protein , interferes with the activation and proliferation of T cells by binding to the P06729 receptor on their surfaces . The clinical efficacy of this drug has been demonstrated in chronic plaque psoriasis . We performed a single-center , open-label study to investigate the immunohistochemical effects in psoriatic lesional skin . A group of 11 patients with plaque psoriasis all received 12 weekly doses of 7.5 mg alefacept intravenously . Skin biopsies were obtained at baseline and on days 8 , 43 and 92 , and were evaluated by digital image analysis after immunohistochemical staining . After completion of treatment , 8 out of the 11 patients experienced a reduction in PASI of 50 % or more compared to baseline . Immunohistochemical analysis displayed a gradual decrease in the number of cutaneous T cells during therapy , with a significant reduction in epidermal CD8+ cells and dermal P01730 + cells on day 92 . Patients with a reduction in PASI of 50 % or more after therapy had a clearance of effector/memory T cells from the epidermis , in contrast to patients with a reduction in PASI of less than 50 % . These findings support the hypothesis that effector/memory T cells play a prominent role in the pathogenesis of psoriasis , and that alefacept is capable of reducing these cells in lesional psoriatic skin . Boosting the sampling efficiency of q-Ball imaging using multiple wavevector fusion . q-Ball imaging ( QBI ) is a high-angular-resolution diffusion imaging ( HARDI ) method that is capable of resolving complex , subvoxel white matter ( WM ) architecture . QBI requires time-intensive sampling of the diffusion signal and large diffusion wavevectors . Here we describe a reconstruction scheme for QBI , termed multiple wavevector fusion ( MWF ) , that substantially boosts the sampling efficiency and signal-to-noise ratio ( SNR ) of QBI . The MWF reconstruction operates by nonlinearly fusing the diffusion signal from separate low and high wavevector acquisitions . The combination of wavevectors provides the benefits of the high SNR of the low wavevector signal and the high angular contrast-to-noise ratio ( P21554 ) and peak separation of the high wavevector signal . The MWF procedure provides a framework for combining diffusion tensor imaging ( DTI ) and QBI . Numerical simulations show that MWF of DTI and QBI provides a more accurate estimate of the diffusion orientation distribution function ( O14788 ) than QBI alone . The accuracy improvement can be translated into an efficiency gain of 274-377 % . An intravoxel peak connectivity metric ( IPCM ) is presented that calculates the peak connectivity between an O14788 and its neighboring voxels . In human WM , MWF reveals more detailed WM architecture than QBI as measured by the IPCM for all sampling schemes presented . DB05258 receptors are important for antiproliferative effect of interferon-alpha against human hepatocellular carcinoma cells . AIM : Interferon ( IFN ) -alpha is a promising drug for the prevention and treatment of hepatocellular carcinoma ( HCC ) . We reported that responders to IFN-alpha/5-fluorouracil combination therapy expressed higher IFN alpha receptor ( P17181 )2 in tumor . Herein we studied involvement of IFNARs in response to IFN-alpha in HCC cells . METHODS : IFN-alpha sensitivity and expression of IFNARs were studied in six HCC cell lines ( HuH7 , P98160 /PRF/5 , P08246 , HLF , HepG2 , Hep3B ) using growth-inhibitory and RT-PCR , Western blot assays . Short interfering RNAs ( SiRNAs ) against P17181 and 2 were used to analyze the role of the IFNARs in IFN-alpha 's effect and signal transduction . RESULTS : The expressions of P17181 and 2c mRNAs were higher in P98160 /PRF/5 cells than those in other cell lines , and P98160 /PRF/5 cells expressed abundant IFNAR2c on their cell membrane . When we examined the sensitivity of the HCC cell lines to the growth-inhibitory effect of IFN-alpha , P98160 /PRF/5 exhibited a significant response , while the other cells were much more resistant . Knockdown of either P17181 or 2 using siRNAs suppressed the IFN-alpha 's signal transduction ( 2.5-fold ) , and decreased the growth-inhibitory effect ( down by 69.9 % and 67.3 % ) . CONCLUSION : The results suggest that the expression of P17181 and IFNAR2c independently are important for the antiproliferative effect of IFN-alpha in HCC cells . Phosphodiesterase-4 inhibitors as a novel approach for the treatment of respiratory disease : cilomilast . Phosphodiesterase-4 ( DB05876 ) is an important DB02527 -metabolising enzyme in immune and inflammatory cells , airway smooth muscle and pulmonary nerves . The phosphodiesterase 4 ( DB05876 ) enzyme plays a significant role in modulating the activity of DB02527 , an important second messenger that mediates the relaxation of airway smooth muscle and suppresses inflammatory cell function , thereby attenuating the inflammatory response . Selective inhibitors of this enzyme show a broad spectrum of activity in animal models of P48444 and asthma . These drugs block the hydrolysis of DB02527 via inhibition of DB05876 and are attractive candidates for novel anti-inflammatory drugs . At present , two second-generation DB05876 inhibitors for the treatment of P48444 and asthma patients are being tested in clinical Phase III trials . The most advanced compound is the orally active , selective DB05876 inhibitor cilomilast ( DB03849 , SB-207499 , cis-4-cyano-4-[3-cyclopentyloxy-4-methoxyphenyl]-cyclohexanecarboxylic acid ; GlaxoSmithKline ) . DB03849 shows high selectivity for DB02527 -specific DB05876 , an isoenzyme that predominates in pro-inflammatory and immune cells and that is 10-fold more selective for Q08499 than for P27815 , -B or -C . In vitro , cilomilast suppresses the activity of several pro-inflammatory and immune cells that have been implicated in the pathogenesis of asthma and P48444 . Moreover , it is highly active in animal models of these diseases . DB03849 has been shown to exert potent anti-inflammatory effects both in vitro and in vivo . It is orally active and may be effective in the treatment of asthma and P48444 ; however , complete assessment of the therapeutic value of this novel compound class must await the outcome of longer-term clinical trials . This review presents a summary of the preclinical and clinical profile of cilomilast in patients with P48444 . The relationship between localized subarachnoid inflammation and parenchymal pathophysiology after spinal cord injury . Subarachnoid inflammation following spinal cord injury ( SCI ) can lead to the formation of localized subarachnoid scarring and the development of post-traumatic syringomyelia ( Q03393 ) . While Q03393 is a devastating complication of SCI , its relative rarity ( occurring symptomatically in about 5 % of clinical cases ) , and lack of fundamental physiological insights , have led us to examine an animal model of traumatic SCI with induced arachnoiditis . We hypothesized that arachnoiditis associated with SCI would potentiate early parenchymal pathophysiology . To test this theory , we examined early spatial pathophysiology in four groups : ( 1 ) sham ( non-injured controls ) , ( 2 ) arachnoiditis ( intrathecal injection of kaolin ) , ( 3 ) SCI ( 35-g clip contusion/compression injury ) , and ( 4 ) Q03393 ( intrathecal kaolin+SCI ) . Overall , there was greater parenchymal inflammation and scarring in the Q03393 group relative to the SCI group . This was demonstrated by significant increases in cytokine ( IL-1α and IL-1β ) and chemokine ( P13500 , P09341 /KC , and MIP-1α ) production , P05164 activity , blood-spinal cord barrier ( BSCB ) permeability , and P14780 activity . However , parenchymal inflammatory mediator production ( acute IL-1α and IL-1β , subacute chemokines ) , BSCB permeability , and fibrous scarring in the Q03393 group were larger than the sum of the SCI group and arachnoiditis group combined , suggesting that arachnoiditis does indeed potentiate parenchymal pathophysiology . Accordingly , these findings suggest that the development of arachnoiditis associated with SCI can lead to an exacerbation of the parenchymal injury , potentially impacting the outcome of this devastating condition . Preclinical to clinical translation of tofacitinib , a Janus kinase inhibitor , in rheumatoid arthritis . A critical piece in the translation of preclinical studies to clinical trials is the determination of dosing regimens that allow maximum therapeutic benefit with minimum toxicity . The preclinical pharmacokinetic ( PK ) /pharmacodynamic ( PD ) profile of tofacitinib , an oral Janus kinase ( JAK ) inhibitor , in a mouse collagen-induced arthritis ( mCIA ) model was compared with clinical PK/PD data from patients with rheumatoid arthritis ( RA ) . Preclinical evaluations included target modulation and PK/PD modeling based on continuous subcutaneous infusion or oral once- or twice-daily ( P55957 ) dosing paradigms in mice . The human PK/PD profile was obtained from pooled data from four phase 2 studies in patients with RA , and maximal effect models were used to evaluate efficacy after 12 weeks of tofacitinib treatment ( 1-15 mg P55957 ) . In mCIA , the main driver of efficacy was inhibition of cytokine receptor signaling mediated by P23458 heterodimers , but not O60674 homodimers , and continuous daily inhibition was not required to maintain efficacy . Projected efficacy could be predicted from total daily exposure irrespective of the oral dosing paradigm , with a total steady-state plasma concentration achieving 50 % of the maximal response ( Cave50 ) of ~100 nM . DB08895 potency ( ED50 ) in clinical studies was ~3.5 mg P55957 ( 90 % confidence interval : 2.3 , 5.5 ) or total Cave50 of ~40 nM , derived using Disease Activity Scores from patients with RA . The collective clinical and preclinical data indicated the importance of Cave as a driver of efficacy , rather than maximum or minimum plasma concentration ( Cmax or Cmin ) , where Cave50 values were within ~2-fold of each other .

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A Nile blue based infrared fluorescent probe : imaging tumors that over-express cyclooxygenase-2 . The first Golgi-localized cyclooxygenase-2 ( P35354 ) -specific near-infrared ( Q9Y3T9 ) fluorescent probe , Niblue- P13671 -IMC , able to detect cancer cells , was designed . Importantly , Niblue- P13671 -IMC preferentially labeled the tumors in a mouse tumor model with deep tissue penetration capacity . It may be a promising molecular tool for guiding tumor resection during surgery . 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 . 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 . 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 . 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 . 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 . Matrix proteinase inhibition by DB05387 , a multifunctional antiangiogenic compound . BACKGROUND : Matrix metalloproteinases ( MMPs ) play an important role in tissue remodeling under normal physiological and pathological conditions and are thus attractive targets for both diagnostic and therapeutic purposes . Here , we examined the effect of DB05387 , an orally bioavailable standardized extract made of cartilage that shows significant antiangiogenic and antimetastatic properties in vivo , on the activity of various members of the MMP family . MATERIALS AND METHODS : The effect of DB05387 on the activity of MMPs was assessed by fluorimetric assays and by substrate gel zymography . RESULTS : DB05387 markedly inhibits the gelatinolytic activity of P08253 and to a lesser extent those of P03956 , P09237 , P14780 and P45452 . DB05387 also inhibited the elastinolytic activities of P08253 and P14780 as well as P39900 ( metalloelastase ) , porcine pancreatic elastase ( PPE ) , and human leukocyte elastase ( P08246 ) . Western blot analysis revealed the presence within DB05387 of immunoreactive P01033 -like proteins , suggesting that these proteins may be at least partly responsible for the observed MMP inhibition . CONCLUSIONS : Taken together , these results demonstrate that DB05387 contains P01033 -like proteins that could be responsible for the specific inhibition of MMPs . Given the recent studies suggesting the presence within this compound of specific inhibitor(s) of endothelial cell proliferation , DB05387 appears as a pleotropic agent able to interfere with several biochemical steps leading to angiogenesis and to other physiopathological conditions . Since DB05387 is currently under Phase III clinical investigations , these findings are also of considerable importance for our understanding of its anticancer properties . DB00945 desensitization : useful treatment for chronic rhinosinusitis with nasal polyps ( CRSwNP ) in aspirin-exacerbated respiratory disease ( AERD ) ? DB00945 intolerance syndrome is due to disturbances in the arachidonic acid metabolism implicating both the lipoxygenase and cyclooxygenase pathways . This results in imbalances of eicosanoid , leukotriene and prostaglandin synthesis . Thus , preinflammatory cysteinyl leukotrienes increase and antiinflammatory prostaglandins ( PG ) such as DB00917 decrease . Clinically , intolerance reactions to nonsteroidal antiinflammatory drugs ( NSAIDs ) can lead to different clinical manifestations ; five phenotypes of the aspirin intolerance syndrome are listed in the ENDA classification . DB00945 -exacerbated respiratory disease ( AERD ) is the most common phenotype characterized by an eosinophil-dominated inflammatory disease of the airways that presents clinically with nasal polyps , chronic sinusitis and bronchial asthma . About 34 % of patients with aspirin-induced asthma and rhinosinusitis are thought to have AERD . Important biochemical findings in many AERD patients are increased basal leukotriene levels ( at least in cell cultures ) that excessively increase after intake of P23219 inhibitors . DB00945 desensitization uses the repetitive application of aspirin to induce a tolerance to NSAIDs , especially P23219 inhibitors . After a dose-increase phase reaching a threshold dose , a dose-continuation phase is performed . For application , the nasal , bronchial , oral and intravenous routes have been described . DB00945 desensitization has been proven to be efficacious and safe and was able to reduce the need for other medications in AERD patients . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . 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 . 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 . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . 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 . 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 . Prevention and treatment of pancreatic cancer by curcumin in combination with omega-3 fatty acids . Pancreatic cancer BxPC-3 cells were exposed to curcumin , docosahexaenoic acid ( DB01708 ) , or combinations of both and analyzed for proliferation and apoptosis . Pancreatic tumor xenografts were established by injecting BxPC-3 cells into each flank of nude mice . After the tumors reached a size of approximately 190-200 mm(3) , animals were fed diets with or without 2,000 ppm curcumin in 18 % corn oil or 15 % fish oil + 3 % corn oil for 6 more wk before assessing the tumor volume and expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygeanse-2 ( P35354 ) , 5-lipoxinase ( 5- P28300 ) , and P38936 . A synergistic effect was observed on induction of apoptosis ( approximately sixfold ) and inhibition of cell proliferation ( approximately 70 % ) when cells were treated with curcumin ( 5 microM ) together with the DB01708 ( 25 microM ) . Mice fed fish oil and curcumin showed a significantly reduced tumor volume , 25 % ( P < 0.04 ) and 43 % ( P < 0.005 ) , respectively , and importantly , a combination of curcumin and fish oil diet showed > 72 % ( P < 0.0001 ) tumor volume reduction . Expression and activity of P35228 , P35354 , and 5- P28300 are downregulated , and P38936 is upregulated in tumor xenograft fed curcumin combined with fish oil diet when compared to individual diets . The preceding results evidence for the first time that curcumin combined with omega-3 fatty acids provide synergistic pancreatic tumor inhibitory properties . Cellular distribution and contribution of cyclooxygenase P35354 to diabetogenesis in NOD mouse . Unlike most other mammalian cells , beta-cells of Langerhans constitutively express cyclooxygenase ( P36551 ) -2 rather than P23219 . P35354 is also constitutively expressed in type 1 diabetes ( T1D ) patients ' periphery blood monocytes and macrophage . To understand the role of P35354 in the beta-cell , we investigated P35354 expression in beta-cells and islet infiltrates of NOD and BALB/c mice using fluorescence immunohistochemistry and cytochemical confocal microscopy and Western blotting . Immunostaining showed that P35354 is expressed in islet-infiltrating macrophages , and that the expression of insulin and P35354 disappeared concomitantly from the beta-cells when NOD mice progressed toward overt diabetes . Also cultured P01308 -1E cells coexpressed insulin and P35354 but clearly in different subcellular compartments . Treatment with celecoxib increased insulin release from these cells in a dose-dependent manner in glucose concentrations ranging from 5 to 17 mM . Excessive P35354 expression by the islet-infiltrating macrophages may contribute to the beta-cell death during insulitis . The effects of celecoxib on P01308 -1E cells suggest that PGE(2) and other downstream products of P35354 may contribute to the regulation of insulin release from the beta-cells . [ Vaccine-associated paralytic poliomyelitis showing biphasic motor paresis ] . We report a 38-year-old man with vaccine associated paralytic poliomyelitis ( VAPP ) which showed unusual biphasic worsening . The patient developed mild paresis of left upper and right lower extremities , five weeks after the oral poliovirus vaccination of patient 's son and two weeks after the intramuscular injection of mumps/varicella vaccine in the left triceps muscle for himself . Needle electromyography ( EMG ) of his left arm and right leg was not remarkable , and the weakness recovered almost completely in three weeks . However , four weeks after the needle EMG , severe weakness and muscle atrophy of the four extremities , accentuated at the left arm and right leg , developed again . Cervical Q9BWK5 showed gadolinium-enhanced , T(2) high-signal intensity area in the left C4- P13671 anterior horn , most prominent at the height of P01031 spine . Significant elevation of serum anti-poliomyelitis type 2 neutralizing antibody confirmed the diagnosis of VAPP . Immunomodulatory treatment , intravenous immunoglobulin ( DB00028 ) , did not improve weakness . We consider that the second clinical worsening of this patient was provoked by the needle EMG performed just after the first exacerbation , which injured the skeletal muscles and might have enhanced the retrograde transport of poliovirus via neural pathway .

MH_train_1440

MH_train_1440

MH_train_1440

interacts_with DB00904?

multiple_choice

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 . 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 . P60568 abolishes myeloid cell accumulation induced by Lewis lung carcinoma . Immune aberration in cancer patients can be at least partly ascribed to an accumulation of immature myeloid cells and monocytes/macrophages with immunosuppressive functions . Mice implanted with Lewis lung carcinoma 2 ( LL/2 ) cells show marked splenomegaly as the tumors progress , and this condition is accompanied by impaired T cell activities . We characterized the cells that accumulated in the spleens of LL/2 tumor-bearing mice and attempted to restore the normal cell population by employing interleukin-2 ( P60568 ) . Flow cytometric analysis revealed that the cells expressing Mac1 , P33681 , NK- P04264 , Gra-1 , and MHC class II antigens on their surfaces drastically decreased in number when LL/2 had been engineered to produce P60568 . P60568 also restored the concanavalin A ( ConA ) -mediated proliferative response and P60568 production of the spleen cells . The in vivo growth of P60568 -producing tumors was significantly slower than that of parental LL/2 cells . Therefore , local P60568 production may reverse systemic immune abnormality by stopping myeloid cell accumulation . 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 . 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 . 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 . NOS Inhibition Modulates Immune Polarization and Improves Radiation-Induced Tumor Growth Delay . DB00435 synthases ( NOS ) are important mediators of progrowth signaling in tumor cells , as they regulate angiogenesis , immune response , and immune-mediated wound healing . Ionizing radiation ( IR ) is also an immune modulator and inducer of wound response . We hypothesized that radiation therapeutic efficacy could be improved by targeting NOS following tumor irradiation . Herein , we show enhanced radiation-induced ( 10 Gy ) tumor growth delay in a syngeneic model ( C3H ) but not immunosuppressed ( Nu/Nu ) squamous cell carcinoma tumor-bearing mice treated post-IR with the constitutive NOS inhibitor N(G)-nitro-l-arginine methyl ester ( L-NAME ) . These results suggest a requirement of T cells for improved radiation tumor response . In support of this observation , tumor irradiation induced a rapid increase in the immunosuppressive Th2 cytokine P22301 , which was abated by post-IR administration of L-NAME . In vivo suppression of P22301 using an antisense P22301 morpholino also extended the tumor growth delay induced by radiation in a manner similar to L-NAME . Further examination of this mechanism in cultured Jurkat T cells revealed L-NAME suppression of IR-induced P22301 expression , which reaccumulated in the presence of exogenous NO donor . In addition to L-NAME , the guanylyl cyclase inhibitors ODQ and thrombospondin-1 also abated IR-induced P22301 expression in Jurkat T cells and Q14201 -1 macrophages , which further suggests that the immunosuppressive effects involve P29474 . Moreover , cytotoxic Th1 cytokines , including P60568 , IL12p40 , and IFNγ , as well as activated CD8(+) T cells were elevated in tumors receiving post-IR L-NAME . Together , these results suggest that post-IR NOS inhibition improves radiation tumor response via Th1 immune polarization within the tumor microenvironment . 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 . 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 . 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 . The anti-interleukin-6 antibody siltuximab down-regulates genes implicated in tumorigenesis in prostate cancer patients from a phase I study . BACKGROUND : P05231 ( P05231 ) is associated with prostate cancer morbidity . In several experimental models , P05231 has been reported to have anti-apoptotic and pro-angiogenic effects . DB09036 ( CNTO 328 ) is a monoclonal anti- P05231 antibody which has been successfully applied in several models representing prostate cancer . This study was designed to assess preliminary safety of siltuximab in patients with early prostate cancer . PATIENTS AND METHODS : Twenty patients scheduled to undergo radical prostatectomy received either no drug or siltuximab ( 6 mg/kg , five patients per group with administration once , two times , and three times prior to surgery ) . Blood samples were collected for pharmacokinetic and pharmacodynamic analyses . Expression of elements of P05231 signaling pathways was analyzed in tumor tissue by immunohistochemistry . Gene analysis in tumor specimens was performed with the DASL array . RESULTS : No adverse events related to siltuximab were observed . Patients treated with siltuximab presented with higher levels of proliferation and apoptosis markers . Following a single dose , serum concentrations of siltuximab declined in a biexponential manner . This study revealed a decrease in phosphorylation of Stat3 and Q8TCB0 / Q8NFH3 mitogen-activated protein kinases . In addition , gene expression analyses indicate down-regulation of genes immediately downstream of the P05231 signaling pathway and key enzymes of the androgen signaling pathway . CONCLUSIONS : Preliminary safety of siltuximab is favorable . Future studies in which siltuximab could be combined with androgen-deprivation therapy and experimental therapies in advanced prostate cancer are justified . The anti-inflammatory and anti-angiogenic role of DB05914 in corneal wound healing following chemical injury . To investigate the anti-inflammatory and anti-angiogenic effects of DB05914 ( O60682 ) in the chemically burned corneas , we mechanically removed the corneal epithelium of rats after 100 % alcohol instillation . The rats were then randomized into four groups : fresh media , conditioned media derived from the O60682 culture ( O60682 -CM ) , O60682 applied topically to the damaged corneas for 2 hours immediately after the injury or O60682 -CM applied either once or 3 times per day for 3 consecutive days . Corneal surface was evaluated every week . After 3 weeks , the corneas were stained with the hematoxylin-eosin , and the expression of interleukin ( IL ) -2 , interferon ( IFN ) -gamma , P05231 , P22301 , transforming growth factor ( TGF ) -beta1 , thrombospondin-1 ( P07996 -1 ) , matrix metalloproteinase-2 ( P08253 ) , and vascular endothelial growth factor ( P15692 ) were analyzed . P01730 + cells were assessed in the corneas . We found that both O60682 and three-time applied O60682 -CM ( 1 ) reduced corneal inflammation and neovascularization , ( 2 ) decreased P60568 and P01579 , although increased P22301 and TGF-beta1 as well as P05231 , ( 3 ) reduced the infiltration of P01730 + cells , and ( 4 ) upregulated the expression of P07996 -1 , although downregulated that of P08253 . Interestingly , whereas three-time application of O60682 -CM was partially effective , transplantation of O60682 achieved a better outcome in suppressing corneal inflammation . The results of this study suggest that the anti-inflammatory and anti-angiogenic action of O60682 in the chemically burned corneas might be mediated in part through paracrine pathways involving soluble factors such as P22301 , TGF-beta1 , P05231 and P07996 -1 . Inhibition of matrix metalloproteinase-2 by halofuginone is mediated by the Egr1 transcription factor . DB04866 , a low-molecular-weight quinazolinone alkaloid that inhibits collagen α1(I) , has been shown to suppress cancer growth , metastasis , and angiogenesis . These activities were attributed in part to the inhibition of matrix metalloproteinase-2 ( P08253 ) . The present study was carried out to explore the molecular mechanism underlying this effect . We found a marked ( 50 % ) inhibition in P08253 gelatinolytic activity in human breast cancer MDA-MB-435 cells pretreated with as little as 50 ng/ml of halofuginone , a concentration that markedly inhibited their invasive and proliferative capacities . We further show that both early growth response 1 ( Egr-1 ) and Nab-2 ( corepressor of Egr1 activation ) are upregulated by halofuginone in a dose-dependent and time-dependent ( up to 5 h ) manner . Using P08253 reporter gene and chromatin immunoprecipitation analyses , we found that Egr-1 binds to the P08253 promoter and inhibits its activity . Altogether , our results identify the downstream elements ( Egr-1 , Nab-2 , and P08253 ) by which halofuginone exerts its antitumoral effect , thereby advancing its potential therapeutic application as an anticancer drug . Modulation by cytokines of glucocorticoid action . Glucocorticoids ( GC ) are potent modulators of the inflammatory response . Their effects serve to down-regulate the inflammatory response and are mediated by genomic pathways that follow the interaction with specific receptors ( glucocorticoid receptors , GR ) . Interleukin ( IL ) -1 , P60568 , and P05231 are able to increase GC secretion by enhancing synthesis and release of P06850 and DB01285 . Cytokine effects upon steroidogenesis also occur at the adrenal level . The role of cytokines as modulators of GR has received scarce attention . IL-1 has been shown to up-regulate GR mRNA expression in hypothalamic P06850 secreting cells . On the other hand , macrophage migration inhibitory factor ( MIF ) , a T-cell product inducible by inflammatory substances including other cytokines , counterregulates GC action within the immune system . Besides immunocytes and neurons , bone cells are a sensitive target for GC and cytokines . We have found that P60568 and P05231 up-regulate remarkably the number of GR binding sites and the expression of GR mRNA in peripheral blood mononuclear cells and in osteoblast-like Saos-2 cells . Available data suggest that inflammatory cytokines have both direct and indirect effects on GC action at the target level . Autocrine-induced transcription of GR in immunocytes and/or osteoblasts could be a mechanism that restrains excess cytokine production . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . 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 .

MH_train_1441

MH_train_1441

MH_train_1441

interacts_with DB01576?

multiple_choice

Adaptive mutations in a human immunodeficiency virus type 1 envelope protein with a truncated V3 loop restore function by improving interactions with P01730 . We previously reported that a human immunodeficiency virus type 1 ( HIV-1 ) clade B envelope protein with a severely truncated V3 loop regained function after passage in tissue culture . The adapted virus , termed Q96RJ0 , retained the V3 truncation , was exquisitely sensitive to neutralization by the P01730 binding site monoclonal antibody b12 and by HIV-positive human sera , used P51681 to enter cells , and was completely resistant to small molecule P51681 antagonists . To examine the mechanistic basis for these properties , we singly and in combination introduced each of the 5 mutations from the adapted clone Q96RJ0 into the unadapted envelope . We found that single amino acid changes in the P01024 region , the V3 loop , and in the fusion peptide were responsible for imparting near-normal levels of envelope function to Q96RJ0 . T342A , which resulted in the loss of a highly conserved glycosylation site in P01024 , played the primary role . The adaptive amino acid changes had no impact on P51681 antagonist resistance but made virus more sensitive to neutralization by antibodies to the P01730 binding site , modestly enhanced affinity for P01730 , and made Q96RJ0 more responsive to P01730 binding . Specifically , Q96RJ0 was triggered by soluble P01730 more readily than the parental Env and , unlike the parental Env , could mediate entry on cells that express low levels of P01730 . In contrast , Q96RJ0 interacted with P51681 less efficiently and was highly sensitive to antibodies that bind to the P51681 N terminus and ECL2 . Therefore , enhanced utilization of P01730 is one mechanism by which HIV-1 can overcome mutations in the V3 region that negatively affect P51681 interactions . Biological characterization of DB05037 , a small-molecule inhibitor of cyclin-dependent kinases , in human tumor cell lines . P12004 -dependent kinases ( CDK ) , and their regulatory cyclin partners , play a central role in eukaryotic cell growth , division , and death . This key role in cell cycle progression , as well as their deregulation in several human cancers , makes them attractive therapeutic targets in oncology . A series of CDK inhibitors was developed using Astex 's fragment-based medicinal chemistry approach , linked to high-throughput X-ray crystallography . A compound from this series , designated DB05037 , is currently in early-phase clinical development . We describe here the biological characterization of DB05037 , a potent inhibitor of several CDK family members . DB05037 showed potent antiproliferative activity ( 40-940 nmol/L ) in a panel of human tumor cell lines , and the mechanism of action was shown here to be consistent with the inhibition of P06493 and P24941 in solid tumor cell lines . DB05037 caused cell cycle arrest followed by apoptosis in human tumor cells and inhibited tumor growth in human tumor xenograft models . Tumor regression was observed following twice daily dosing of DB05037 in the HCT116 and HT29 colon cancer xenograft models . We show that these biological effects are linked to inhibition of CDKs in vivo and that DB05037 induces tumor cell apoptosis in these xenograft models . DB05037 has an attractive biological profile for development as a clinical candidate , and the tolerability and efficacy in animal models compare favorably with other CDK inhibitors in clinical development . Studies described here formed the biological rationale for investigating the potential therapeutic benefit of DB05037 in cancer patients . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . 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 . Dopamine modulating drugs influence striatal (+)-[11C]DTBZ binding in rats : Q05940 binding is sensitive to changes in vesicular dopamine concentration . Binding of (+)-[11C]DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of (+)-[11C]DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , +14 % ) or DB01576 ( d- P49418 , 20 mg/kg , +12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , -16 % ) or levodopa ( -20 % ) . We suggest that in vivo (+)-[11C]DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed . 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 . DB06288 promotes cognitive flexibility in rats : the role of P34969 receptors . The antagonism of P34969 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug , amisulpride . It has been previously demonstrated that the selective P34969 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task ( ASST ) . Therefore , the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility . The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound 's action at P34969 receptors . Rats repeatedly exposed ( 1 h daily for 7 days ) to restraint stress demonstrated impaired performance on the extra-dimensional ( ED ) set-shifting stage of the ASST . DB06288 ( 3 mg/kg ) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group . The P34969 receptor agonist , AS19 ( 10 mg/kg ) , abolished the pro-cognitive efficacy of amisulpride ( 3 mg/kg ) . The present study suggests that the antagonism of P34969 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride . These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders . Pathogenesis and treatment of thrombohemorrhagic diathesis in acute promyelocytic leukemia . Acute promyelocytic leukemia ( APL ) is a distinct subtype of myeloid leukemia characterized by t(15;17) chromosomal translocation , which involves the retinoic acid receptor-alpha ( P10276 ) . APL typically presents with a life-threatening hemorrhagic diathesis . Before the introduction of all-trans retinoic acid ( DB00755 ) for the cure of APL , fatal hemorrhages due , at least in part , to the APL-associated coagulopathy , were a major cause of induction remission failure . The laboratory abnormalities of blood coagulation found in these patients indicate the occurrence of a hypercoagulable state . Major determinants of the coagulopathy of APL are endogenous factors expressed by the leukemic cells , including procoagulant factors , fibrinolytic proteins , and non-specific proteolytic enzymes . In addition , these cells have an increased capacity to adhere to the vascular endothelium , and to secrete inflammatory cytokines [ i.e. interleukin-1beta ( IL-1beta ) and tumor necrosis factor ( P01375 ) ] , which in turn stimulate the expression of prothrombotic activities by endothelial cells and leukocytes . DB00755 can interfere with each of the principal hemostatic properties of the leukemic cell , thus reducing the APL cell procoagulant potential , in parallel to the induction of cellular differentiation . This effect occurs in vivo , in the bone marrow of APL patients receiving DB00755 , and is associated with the improvement of the bleeding symptoms . Therapy with arsenic trioxide ( ATO ) also beneficially affects coagulation in APL . However , early deaths from bleeding still remain a major problem in APL and further research is required in this field . In this review , we will summarize our current knowledge of the pathogenesis of the APL-associated coagulopathy and will overview the therapeutic approaches for the management of this complication . 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 . DNA-based Q9BWK5 probes for specific detection of chronic exposure to amphetamine in living brains . We designed phosphorothioate-modified DNA probes linked to superparamagnetic iron oxide nanoparticles ( SPION ) for in vivo magnetic resonance imaging ( Q9BWK5 ) of fosB and Delta fosB mRNA after amphetamine ( P49418 ) exposure in mice . Specificity of both the fosB and Delta fosB probes was verified by in vitro reverse transcriptase-PCR amplification to a single fragment of total cDNA obtained from acutely P49418 -exposed mouse brains . We confirmed time-dependent uptake and retention profiles of both probes in neurons of Q99259 -green fluorescent protein knock-in mice . Q9BWK5 signal of SPION-labeled fosB probe delivered via intracerebroventricular route was elevated in both acutely and chronically P49418 -exposed mice ; the signal was suppressed by dopaminergic receptor antagonist pretreatment . SPION-labeled Delta fosB probe signal elevation occurred only in chronically P49418 -exposed mice . The in vivo target specificity of these probes permits reliable Q9BWK5 visualization of P49418 -induced differential elevations of fosB and Delta fosB mRNA in living brains . 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 . Cdk5/p25(nck5a) interaction with synaptic proteins in bovine brain . P12004 -dependent kinase 5 ( Cdk5 ) exists in large multimeric complexes , but its function and binding partners in these complexes are unclear . We explored these issues by chromatographic and immunochemical analyses of Cdk5 and p25(nck5a) ( a neuronal Cdk5 activator ) and their associated proteins from bovine brain . Mono-S column enzyme eluates were divided into three fractions and analyzed by gel filtration . The majority of p25(nck5a) from Mono-S fractions I , II , and III eluted from the gel filtration column at approximately 60 , 200 , and 400 kDa , respectively , and Cdk5 was abundant in fractions > 400 kDa . We characterized these macromolecular structures by immunoprecipitating p25(nck5a) , followed by a second immunoprecipitation of remaining unbound proteins using a Cdk5 antibody . The p25(nck5a) immunoprecipitates showed association with Cdk5 . P49418 was detected in the 400-kDa complex and synapsin I in the > 400 kDa structure . The Cdk5 immunoprecipitates , however , revealed abundant retained Cdk5 but no remaining p25(nck5a) , indicating that Cdk5 in macromolecular structures is mostly unassociated with p25(nck5a) . Thus , we demonstrate : an amphiphysin-associated 400-kDa Cdk5/p25(nck5a) complex , a synapsin I-associated > 400-kDa Cdk5/p25(nck5a) complex , and nck5a-free Cdk5 complexes ( 200 to > 400 kDa ) . P49418 acts as a Cdk5/p25(nck5a) substrate in the 400-kDa complex and we speculate that Cdk5/p25(nck5a) participates in amphiphysin-mediated endocytosis . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . 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 . Heterodimerization of human apelin and bradykinin 1 receptors : novel signal transduction characteristics . P35414 ( P35414 ) and bradykinin 1 receptor ( P46663 ) are involved in a variety of important physiological processes , which share many similar characteristics in distribution and functions in the cardiovascular system . This study explored the possibility of heterodimerization between P35414 and P46663 , and investigated the impact of heterodimer on the signal transduction characteristics and the physiological functions in human endothelial cells after stimulation with their agonists . We first identified the endogenous expression of P35414 and P46663 in HUVECs and their co-localization on HEK293 membrane . The constitutive heterodimerization between the P35414 and P46663 was then demonstrated by BRET and FRET assays . Stimulation with Q9ULZ1 -13 and des - DB00125 (9)-BK enhanced the phosphorylation of P29474 in HUVECs , which could be dampened by the knockdown of P35414 or P46663 , indicating the co-existence of P35414 and P46663 is critical for P29474 phosphorylation in HUVECs . Furthermore , P35414 / P46663 heterodimers were found to enhance the activity of PKC signaling pathway and increase intracellular Ca(2+) concentration in HEK293 cells , which might be the mechanism of P35414 / P46663 heterodimers promoting the phosphorylation of P29474 and leads to increased Gαq , PKC signal pathway activities and a significant increase in cell proliferation . The results provide a new theoretical and experimental base for revealed intracellular molecular mechanisms of physiological function involved in the P35414 and P46663 and provide potential new targets for the development of drugs and treating cardiovascular disease . 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 . Large candidate gene association study reveals genetic risk factors and therapeutic targets for fibromyalgia . OBJECTIVE : Fibromyalgia ( FM ) represents a complex disorder that is characterized by widespread pain and tenderness and is frequently accompanied by additional somatic and cognitive/affective symptoms . Genetic risk factors are known to contribute to the etiology of the syndrome . The aim of this study was to examine > 350 genes for association with FM , using a large-scale candidate gene approach . METHODS : The study group comprised 496 patients with FM ( cases ) and 348 individuals with no chronic pain ( controls ) . Genotyping was performed using a dedicated gene array chip , the Pain Research Panel , which assays variants characterizing > 350 genes known to be involved in the biologic pathways relevant to nociception , inflammation , and mood . Association testing was performed using logistic regression . RESULTS : Significant differences in allele frequencies between cases and controls were observed for 3 genes : P28472 ( rs4906902 ; P = 3.65 × 10(-6) ) , Q96RJ0 ( rs8192619 ; P = 1.11 × 10(-5) ) , and P32455 ( rs7911 ; P = 1.06 × 10(-4) ) . These 3 genes and 7 other genes with suggestive evidence for association were examined in a second , independent cohort of patients with FM and control subjects who were genotyped using the Perlegen 600K platform . Evidence of association in the replication cohort was observed for Q96RJ0 , P49798 , P21554 , and P48058 . CONCLUSION : Variation in these 4 replicated genes may serve as a basis for development of new diagnostic approaches , and the products of these genes may contribute to the pathophysiology of FM and represent potential targets for therapeutic action . Stability of neuronal and glial marker enzymes in post-mortem rat brain . The enzymatic activities in post-mortem rat brain kept at 4 degrees C and at 25 degrees C were determined for a number of enzymes localized in specific cell types in the central nervous system . Choline acetyltransferase ( CAT ) , glycerol-3-phosphate dehydrogenase ( GPDH ) , glutamine synthetase ( GS ) , lactate dehydrogenase ( LDH ) and 2',3'-cyclic nucleotide phosphohydrolase ( P09543 ) were found to be very stable at both 4 degrees C and 25 degrees C with only slight , if any , losses of activity being seen even at periods as long as 72 hr . DB00142 decarboxylase ( Q99259 ) activity was less stable than that of the other enzymes . In brains kept at 4 degrees C Q99259 activity was stable out to 24 hr after which it began to decline rapidly to 65 % of control at 72 hr . In brains kept at 25 degrees C , Q99259 activity was stable for 6-8 hr and then began to steadily decline to 58 % of control at 24 hr and 29 % of control at 72 hr . Assuming that these enzymes have similar stabilities in post-mortem human brain , the effect of post-mortem delay in processing tissues may be of lesser significance than other factors with regard to the measured enzyme activities in human brain samples . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . Effects of antihistamines on the function of human α7-nicotinic acetylcholine receptors . Effects of the histamine H₁ receptor ( P35367 ) antagonists ( antihistamines ) , promethazine ( PMZ ) , orphenadrine ( ORP ) , chlorpheniramine ( CLP ) , DB06691 ( PYR ) , diphenhydramine ( DPH ) , citerizine ( CTZ ) , and triprolidine ( TRP ) on the functional properties of the cloned α7 subunit of the human nicotinic acetylcholine receptor expressed in Xenopus oocytes were investigated . Antihistamines inhibited the α7-nicotinic acetylcholine receptor in the order PYR > CLP > TRP > PMZ > ORP≥DPH≥CTZ . Among the antihistamines , PYR showed the highest reversible inhibition of acetylcholine ( 100 µM ) -induced responses with IC₅₀ of 6.2 µM . PYR-induced inhibition was independent of the membrane potential and could not be reversed by increasing the concentration of acetylcholine . Specific binding of [ ¹²⁵I ] α-bungarotoxin , a selective antagonist for α7-nicotinic acetylcholine receptor , was not changed in the presence of PYR suggesting a non-competitive inhibition of nicotinic receptors . In line with functional experiments , docking studies indicated that PYR can potentially bind allosterically with the α7 transmembrane domain . Our results indicate that the H₂-H₄ receptor antagonists tested in this study ( 10 µM ) showed negligible inhibition of α7-nicotinic acetylcholine receptors . On the other hand , H₁ receptor antagonists inhibited the function of human α7-nicotinic acetylcholine receptor , with varying potencies . These results emphasize the importance of α7-nicotinic acetylcholine receptor for future pharmacological/toxicological profiling . 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 . 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 .

MH_train_1442

MH_train_1442

MH_train_1442

interacts_with DB01151?

multiple_choice

27-hydroxycholesterol is an endogenous ligand for liver X receptor in cholesterol-loaded cells . The nuclear receptors liver X receptor alpha ( LXRalpha ) ( Q13133 ) and LXRbeta ( P55055 ) are important regulators of genes involved in lipid metabolism , including O95477 , P45844 , and sterol regulatory element-binding protein-1c ( SREBP-1c ) . Although it has been demonstrated that oxysterols are LXR ligands , little is known about the identity of the physiological activators of these receptors . Here we confirm earlier studies demonstrating a dose-dependent induction of O95477 and P45844 in human monocyte-derived macrophages by cholesterol loading . In addition , we show that formation of 27-hydroxycholesterol and cholestenoic acid , products of Q02318 action on cholesterol , is dependent on the dose of cholesterol used to load the cells . Other proposed LXR ligands , including 20(S)-hydroxycholesterol , 22(R)-hydroxycholesterol , and 24(S),25-epoxycholesterol , could not be detected under these conditions . A role for Q02318 in regulation of cholesterol-induced genes was demonstrated by the following findings . 1 ) Introduction of Q02318 into P29320 -293 cells conferred an induction of P45844 and SREBP-1c ; 2 ) upon cholesterol loading , Q02318 -expressing cells induce these genes to a greater extent than in control cells ; 3 ) in Q02318 -deficient human skin fibroblasts , the induction of O95477 in response to cholesterol loading was ablated ; and 4 ) in a coactivator association assay , 27-hydroxycholesterol functionally activated LXR . We conclude that 27-hydroxylation of cholesterol is an important pathway for LXR activation in response to cholesterol overload . 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 . 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 . Battle of the kinases : integration of adrenal responses to DB02527 , DG and Ca2+ at the level of steroidogenic cytochromes P450 and 3betaHSD expression in H295R cells . While DB01285 receptors ( activating the protein kinase A pathway ) are expressed throughout the human/bovine/ovine zona glomerulosa ( zg ) and zona fasciculata ( zf ) , there are clear zonal differences in AII Type-1 receptor levels ( activating protein kinase C/Ca2+ ) , as well as resting membrane potential . Thus zg is most responsive to AII and K+ ( Ca2+ signalling ) , while zf is less responsive to AII with no K+ response . Zonal function in turn requires differential expression of P05093 /3betaHSD and P19099 / P15538 . We have used the H295R cell to study how differential activation of kinase A , kinase C and Ca2+/calmodulin ( P62158 ) kinases may alter the relative expression of the steroidogenic P450s and 3betaHSDII . While P05108 , P05093 , 3betaHSDII , P08686 , and P15538 are all induced by increases in DB02527 , studies with TPA alone or in combination with forskolin reveal subsets of steroidogenic enzymes regulated either positively ( P08686 , 3betaHSDII ) or negatively ( P05093 , P05108 ) by protein kinase C . Thus adrenal 3betaHSDII and P08686 expression is high in zg and zf , but P05093 is not expressed in the zg where AII activation of kinase C is highest . In turn both K+ and AII-induced elevation of Ca2+ strongly induces P19099 but not P15538 , consistent with preferential expression of P19099 in the zg . We conclude that differential signaling through kinase C and P62158 kinases in addition to kinase A underlies zonal differences in both the early and late pathways involved in steroid hormone production within the adrenocortical zones . 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 . 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 . 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 . 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 . 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 . On the specific interaction between the lysine-binding sites in plasmin and complementary sites in alpha2-antiplasmin and in fibrinogen . P00747 and plasminogen derivatives which contain lysine-binding sites were found to decrease the reaction rate between plasmin and alpha2-antiplasmin by competing with plasmin for the complementary site(s) in alpha2-antiplasmin . The dissocwation constant Kd for the interaction between intact plasminogen ( DB00142 -plasminogen ) and alpha2-antiplasmin is 4.0 microM but those for Lys-plasminogen or TLCK-plasmin are about 10-fold lower indicating a stronger interaction . The lysine-binding site(s) which is situated in triple-loops 1 -- 3 in the plasmin A-chain is mainly responsible for the interaction with alpha2-antiplasmin . The interaction between DB00142 -plasminogen and alpha2-antiplasmin furthermore enhances the activation of DB00142 -plasminogen by urokinase to a comparable extent as DB00513 , suggesting that similar conformational changes occur in the proenzyme after complex formation . DB09222 , fibrinogen digested with plasmin , purified fragment E and purified fragment D interfere with the reaction between plasmin and alpha2-antiplasmin by competing with alpha2-antiplasmin for the lysine-binding site(s) in the plasmin A-chain . The Kd obtained for these interactions varied between 0.2 microM and 1.4 microM ; fragment E being the most effective . Thus the fibrinogen molecule contains several complementary sites to the lysine-binding sites located both in its NH2-terminal and COOH-terminal regions ; these sites are to a large extent . Protective effects of fermented ginseng on streptozotocin-induced pancreatic beta-cell damage through inhibition of NF-kappaB . DB01404 ( Panax ginseng C.A. Meyer ) is widely used in Asian countries as a traditional medicine for the treatment of various diseases . It is known to have anti-inflammatory effects , although the mechanism is not clear . In this study , preventive effects of fermented ginseng ( FG ) against streptozotocin ( Q11206 ) -induced pancreatic beta-cell death was assessed in RINm5F insulinoma cells . FG markedly inhibited the production of nitrite in a dose-dependent manner . The decrease in nitrite production was found to correlate with reduced inducible nitric oxide ( P35228 ) protein and mRNA levels . To characterize the anti-inflammatory mechanism of FG at the transcriptional level , we examined effects of FG on the activity of nuclear factor-kappaB ( NF-kappaB ) . FG reduced a translocation of the NF-kappaB subunit and NF-kappaB-dependent transcriptional activity . FG blocked signaling upstream of NF-kappaB activation , such as degradation of inhibitor factor-kappaBalpha ( P25963 ) and phosphorylations of extracellular signal-regulated kinase ( P29323 ) and c-Jun NH2-terminal kinase ( JNK ) . These results suggest that FG protects against Q11206 -induced pancreatic beta-cell damage by downregulation of P35228 , cyclooxygenase-2 ( P35354 ) , and tumor necrosis factor-alpha ( P01375 ) gene expressions by blocking NF-kappaB and mitogen-activated protein kinase activities . 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 . 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 . P22888 disorder in endometriosis . DB00044 ( LH ) receptor concentrations in ovarian follicles and corpora lutea were measured in 51 patients with histologically proven endometriosis and in 41 control patients . The LH receptor concentrations in cases of endometriosis were lower during the early ( 0.43 +/- 0.11 [ mean +/- standard error ] versus 1.31 +/- 0.27 fmol/mg protein ; P less than 0.001 ) and late ( 0.48 +/- 0.10 versus 1.59 +/- 0.22 fmol/mg protein ; P less than 0.001 ) follicular phase , and during the late luteal phase ( 2.62 +/- 0.55 versus 4.62 +/- 0.65 fmol/mg protein ; P less than 0.05 ) of the cycle than in control patients . In contrast to the control patients , the LH receptor concentration during the follicular phase remained constant in endometriosis , being lower in patients with extensive or severe disease than in patients with moderate or mild disease ( 0.28 +/- 0.07 versus 0.61 +/- 0.21 fmol/mg protein ; P less than 0.05 ) . Endometriosis-associated infertility might be a consequence of a defect in the mechanism mediating LH action in the ovaries . Antitumor activity with P05093 blockade indicates that castration-resistant prostate cancer frequently remains hormone driven . DB05812 acetate is a potent , selective , and orally bioavailable small molecule inhibitor of P05093 , an enzyme that catalyzes two key serial reactions ( 17 alpha hydroxylase and 17,20 lyase ) in androgen and estrogen biosynthesis . Clinical trials have confirmed that specific inhibition of P05093 is safe and results in clinically important antitumor activity in up to 70 % of castrate patients with advanced prostate cancer resistant to currently available endocrine therapies . These clinical data indicate that castration-resistant prostate cancer frequently remains hormone dependent and has confirmed that this disease should no longer be described as " hormone resistant or refractory " . Biomarker studies , including the analysis of ETS gene fusion status , on patients treated with abiraterone acetate may allow enrichment of patients with a sensitive phenotype in future studies of therapeutics targeting P05093 . 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 . Red cell and serum protein polymorphisms in three population groups of South Korea . Genetic markers P24666 , P36871 with subtypes , P10768 , Q04760 , P52209 , GPT , A6NDG6 , P01024 , TF and GC with subtypes , BF , HP , Q9BXS0 , P00747 and PI , were studied in three populations in South Korea , one being the population of the industrial capital Seoul , the second a rural group from Taejon and the third the population of Cheju Island . For the polymorphic systems studied in the present work , a general similarity was observed among the three populations , with the exception of GPT and P24666 ( Taejon vs. Seoul ) and subtypes of GC ( Taejon vs. Cheiu ) . 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 . 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 . 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 . DB08911 : first global approval . DB08911 is an orally bioavailable mitogen-activated protein kinase ( MAPK ) kinase ( MEK ) inhibitor with antineoplastic activity . The compound specifically binds to Q02750 and P36507 , resulting in inhibition of growth factor-mediated cell signalling and cellular proliferation in various cancers . Originally developed by Japan Tobacco , GlaxoSmithKline has licensed exclusive worldwide rights to the compound and conducted development in a number of different cancer types . DB08911 , as a monotherapy , has been approved in the US for the treatment of unresectable or metastatic malignant melanoma with P15056 V600E or V600K mutations , as detected by an FDA-approved test . The compound , as a monotherapy , has also been submitted for regulatory review in the EU for P15056 mutation-positive malignant melanoma , and is in phase III development in Europe , Argentina , Canada and Oceania . Phase II development is underway for pancreatic cancer , non-small cell lung cancer and relapsed or refractory leukaemias . GlaxoSmithKline is also developing trametinib for use in combination with dabrafenib in P15056 V600 mutation-positive metastatic cutaneous melanoma ; the combination is at the preregistration stage in the EU and a phase III clinical programme is underway worldwide . Phase II development for this combination is also underway in colorectal cancer . Several phase I trials have also been initiated to evaluate trametinib in combination with other drugs for the treatment of various solid tumours and haematological malignancies . A paediatric oral solution formulation has been assessed against the oral tablet formulation in a phase I trial . This article summarizes the milestones in the development of trametinib leading to this first approval for unresectable or metastatic P15056 mutation-positive malignant melanoma .

MH_train_1443

MH_train_1443

MH_train_1443

interacts_with DB01109?

multiple_choice

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 . 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 . Phosphorylation of beta-arrestin2 regulates its function in internalization of beta(2)-adrenergic receptors . Beta-arrestins mediate agonist-dependent desensitization and internalization of G protein-coupled receptors . Previously , we have shown that phosphorylation of beta-arrestin1 by ERKs at DB00133 -412 regulates its association with clathrin and its function in promoting clathrin-mediated internalization of the receptor . In this paper we report that beta-arrestin2 is also phosphorylated , predominantly at residues DB00156 -383 and DB00133 -361 . DB01064 stimulation of the beta(2)-adrenergic receptor promotes dephosphorylation of beta-arrestin2 . Mutation of beta-arrestin2 phosphorylation sites to aspartic acid decreases the association of beta-arrestin2 with clathrin , thereby reducing its ability to promote internalization of the beta(2)-adrenergic receptor . Its ability to bind and desensitize the beta(2)-adrenergic receptor is , however , unaltered . These results suggest that , analogous to beta-arrestin1 , phosphorylation/dephosphorylation of beta-arrestin2 regulates clathrin-mediated internalization of the beta(2)-adrenergic receptor . In contrast to beta-arrestin1 , which is phosphorylated by P27361 and P28482 , phosphorylation of beta-arrestin2 at DB00156 -383 is shown to be mediated by casein kinase II . Recently , it has been reported that phosphorylation of visual arrestin at DB00133 -366 prevents its binding to clathrin . Thus it appears that the function of all arrestin family members in mediating internalization of G protein-coupled receptors is regulated by distinct phosphorylation/dephosphorylation mechanisms . 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 ) . 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 . The cell adhesion protein P16109 glycoprotein ligand-1 is a substrate for the aspartyl protease P56817 . The aspartyl protease P56817 cleaves the amyloid precursor protein and the sialyltransferase P15907 and is important in the pathogenesis of Alzheimer 's disease . The normal function of P56817 and additional physiological substrates have not been identified . Here we show that P56817 acts on the Q14242 ( Q14242 ) , which mediates leukocyte adhesion in inflammatory reactions . In human monocytic U937 and human embryonic kidney 293 cells expressing endogenous or transfected P56817 , Q14242 was cleaved by P56817 to generate a soluble ectodomain and a C-terminal transmembrane fragment . No evidence of the cleavage fragment was seen in primary cells derived from mice deficient in P56817 . By using deletion constructs and enzymatic deglycosylation of the C-terminal Q14242 fragments , the cleavage site in Q14242 was mapped to the juxtamembrane region within the ectodomain . In an in vitro assay P56817 catalyzed the formation of the Q14242 products seen in vivo . The cleavage occurred at a DB00149 - DB00133 peptide bond as identified by mass spectrometry using a synthetic peptide . We conclude that Q14242 is an additional substrate for P56817 . Mapping four genes from human chromosome 4 to porcine chromosome 8 further develops the comparative map for an economically important chromosome of the swine genome . Because porcine chromosome ( SSC ) 8 has become the focal point of many efforts aimed at identifying quantitative trait loci affecting ovulation rate , genes distributed across human chromosome ( HSA ) 4 were physically mapped in the pig . A more refined comparative map of this region for these two species was produced . In this study , four genes were selected based on their location in the human genome , the availability of nucleotide sequence and their genomic organization . The genes selected were fibroblast growth factor basic ( P09038 ; HSA 4q25-27 ) , gonadotropin releasing hormone receptor ( P30968 ; HSA 4q13 ) , phosphodiesterase 6 B ( P35913 ; HSA 4p16.3 ) and aminopeptidase S ( P28838 ; HSA 4p11-q12 ) . Genomic libraries were screened via PCR and clones were physically assigned using fluorescence in situ hybridization ( Q5TCZ1 ) . These four genes from HSA 4 were physically mapped to SSC 8p2.3 ( P35913 ) , 8p1.1 ( P28838 ) , 8q1.1-1.2 ( P30968 ) and 8q2.2-2.4 ( P09038 ) . These assignments provide additional benchmarks for the comparative map and help define the level of gene order conserved between HSA 4 and SSC 8 . 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 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 . Q14242 and P42345 regulate translation of ROCK-1 and physiological functions of macrophages . Rho-associated kinases ( ROCKs ) are critical molecules involved in the physiological functions of macrophages , such as chemotaxis and phagocytosis . We demonstrate that macrophage adherence promotes rapid changes in physiological functions that depend on translational upregulation of preformed ROCK-1 mRNA , but not ROCK-2 mRNA . Before adherence , both ROCK mRNAs were present in the cytoplasm of macrophages , whereas ROCK proteins were undetectable . Macrophage adherence promoted signaling through P16109 glycoprotein ligand-1 ( Q14242 ) /Akt/ P42345 that resulted in synthesis of ROCK-1 , but not ROCK-2 . Following synthesis , ROCK-1 was catalytically active . In addition , there was a rapamycin/sirolimus-sensitive enhanced loading of ribosomes on preformed ROCK-1 mRNAs . Inhibition of P42345 by rapamycin abolished ROCK-1 synthesis in macrophages resulting in an inhibition of chemotaxis and phagocytosis . Macrophages from Q14242 -deficient mice recapitulated pharmacological inhibitor studies . These results indicate that receptor-mediated regulation at the level of translation is a component of a rapid set of mechanisms required to direct the macrophage phenotype upon adherence and suggest a mechanism for the immunosuppressive and anti-inflammatory effects of rapamycin/sirolimus . DB00741 does not mediate the suppressive effects of psychiatric morbidity on natural killer cell activity : a cross-sectional study of patients with early breast cancer . BACKGROUND : There is evidence that depression impairs natural killer cell activity ( P20366 ) ; this could have implications for anti-tumour immunity . Our aim was to examine the role of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis in suppressing P20366 in a population of patients with early breast cancer , screened for depression . Secondary aims were to study the relationship between psychological , endocrine and immune variables and baseline tumour characteristics . METHOD : A cross-sectional population of female patients ( n=55 ) with early breast cancer was sampled prior to primary surgery . Structured interview and psychometric instruments measured psychological distress . Flow cytometry was used to enumerate NK cells and lymphocytes were cryopreserved for use in a 51Cr-release assay , to estimate P20366 . Midnight and three early morning saliva samples were collected to measure free cortisol levels . Tumour characteristics were obtained from hospital laboratory data . RESULTS : A high rate of psychological morbidity ( 40 % ) was observed in the population . P20366 was reduced in those with past or current psychiatric illness compared to those without ( 344 v. 553 LU20 and 455 v. 569 LU20 respectively , p < 0.05 for both ) . DB00741 was not related to psychological status but was modestly positively correlated to P20366 . A positive correlation was observed between the Fighting Spirit subscale of the Mental Adjustment to Cancer Scale and tumour size ( r=0.383 , p=0.012 ) . CONCLUSIONS : Our data support the evidence that psychological morbidity is associated with immune dysfunction ; however , the most obvious candidate mediator of this effect , the Q9Y251 axis , does not appear responsible for this effect . Possible reasons for this are discussed . Inhibition of HCV by the serpin antithrombin III . BACKGROUND : Although there have been dramatic strides made recently in the treatment of chronic hepatitis C virus infection , interferon-α based therapy remains challenging for certain populations , including those with unfavorable Q8IZI9 genotypes , psychiatric co-morbidity , HIV co-infection , and decompensated liver disease . We have recently shown that P01008 , a serine protease inhibitor ( serpin ) , has broad antiviral properties . RESULTS : We now show that P01008 is capable of inhibiting HCV in the OR6 replicon model at micromolar concentrations . At a mechanistic level using gene-expression arrays , we found that P01008 treatment down-regulated multiple host cell signal transduction factors involved in the pathogenesis of cirrhosis and hepatocellular carcinoma , including Jun , Myc and P12643 . Using a protein interactive network analysis we found that changes in gene-expression caused by P01008 were dependent on three nodes previously implicated in HCV disease progression or HCV replication : NFκB , O75791 MAPK , and P27361 /2 . CONCLUSIONS : Our findings suggest that P01008 stimulates a novel innate antiviral host cell defense different from current treatment options . gamma-Glutamyltranspeptidase-conferred resistance to hydroquinone induced DB00143 depletion and toxicity in isolated hepatocytes . Hepatocyte resistance against glutathione ( DB00143 ) depleting xenobiotics was studied in an in vitro model . Hepatocytes were isolated from carcinogen treated rats that had received phenobarbital for three weeks . Isolated cells were incubated in DB00143 containing buffer with hydroquinone , which depleted DB00143 . Cells were then seeded on collagen coated plates and cultured overnight in complete medium . Attached cells were stained and the proportion of gamma-glutamyltranspeptidase ( P19440 ) -positive cells was counted . It was found that toxicity related to DB00143 depletion increased the proportion of P19440 -positive cells from 10-15 % up to 40-60 % , indicating that the toxicity mainly affected P19440 -negative cells . DB00143 added to the buffer was essential for this effect . It is concluded that P19440 may protect P19440 -positive hepatocytes from DB00143 depletion and toxicity early during liver carcinogenesis . 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 . Clinical significance of arginase after liver transplantation . Liver graft function after transplantation is dependent on ischemia-reperfusion injury , toxicity of drugs ( immunosuppression , antibiotics and other ) and transplant rejection . Although routinely monitored with enzymatic tests ( Q9NRA2 , ALT , P19440 , ALP ) , bilirubin and coagulation parameters , differentiation between these pathologies is hardly possible without liver biopsy . Arginase ( 3.5.3.1 ) mostly exists in the liver and in trace amounts in extra-hepatic tissue . Thus , we hypothesized that activity of arginase could be a more specific test of liver function . Sera of 32 liver transplant recipients were tested for Q9NRA2 , ALT , P01008 , bilirubin and arginase . Samples were obtained daily in first 2 weeks after LTx and weekly afterwards . Correlation of arginase activity with other liver function markers was calculated . Serum arginase peaked at day 1 post LTx ( mean 64,6+/-91 IU/L ) , and decreased more rapidly than other tests if good liver function was observed . The values showed strong and significant correlation with Q9NRA2 and ALT activities ( Pearsons R 0,65 and 0,47 respectively ) . We conclude that activity of arginase in the serum is an exact test of liver function . 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 . 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 . 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 . [ 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 . P10275 expressing neurons that project to the paraventricular nucleus of the hypothalamus in the male rat . Androgen receptors are distributed throughout the central nervous system and are contained by a variety of nuclei that are known to project to or regulate the paraventricular nucleus ( PVN ) of the hypothalamus , the final common pathway by which the brain regulates the hypothalamic-pituitary-adrenal ( Q9Y251 ) response to homeostatic threat . Here we characterized androgen receptor staining within cells identified as projecting to the PVN in male rats bearing iontophoretic or crystalline injections of the retrograde tracer FluoroGold aimed at the caudal two-thirds of the nucleus , where corticotropin-releasing hormone-expressing neurons are amassed . P10275 ( AR ) and FluoroGold ( FG ) double labeling was revealed throughout the limbic forebrain , including scattered numbers of cells within the anterior and posterior subdivisions of the bed nuclei of the stria terminalis ; the medial zone of the hypothalamus , including large numbers of AR-FG-positive cells within the anteroventral periventricular and medial preoptic cell groups . Strong and consistent colabeling was also revealed throughout the hindbrain , predominantly within the periaqueductal gray and the lateral parabrachial nucleus , and within various medullary cell groups identified as catecholaminergic , predominantly C1 and A1 neurons of the ventral medulla . These connectional data predict that androgens can act on a large assortment of multimodal inputs to the PVN , including those involved with the processing of various types of sensory and limbic information , and provide an anatomical framework for understanding how gonadal status could contribute to individual differences in Q9Y251 function . 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 .

MH_train_1444

MH_train_1444

MH_train_1444

interacts_with DB02901?

multiple_choice

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 . Parathyroid hormone activates PKC-delta and regulates osteoblastic differentiation via a P98160 -independent pathway . PTH exerts major effects upon bone by activating PTH/ P12272 receptors ( PTH1Rs ) expressed on osteoblasts . The Q03431 is capable of engaging multiple signaling pathways in parallel , including Gs/adenylyl cyclase ( AC ) , Gq/phospholipase C/protein kinase C ( P98160 /PKC ) and a distinct mechanism , involving activation of PKC via a P98160 -independent pathway , that depends upon ligand determinants within the PTH(29-34) sequence . The involvement of P98160 -dependent vs. P98160 -independent PKC activation in PTH action was studied in clonal Q03431 -expressing murine calvarial osteoblasts ( " Wt9 " ) using two signal-selective analogs , [ P55008 ,R19] DB05829 (1-28) and [ P55008 ,R19] DB05829 (1-34) . Both analogs lack P98160 signaling but differ in their capacity to activate the P98160 -independent PKC pathway . Both DB05829 (1-34) and [ P55008 ,R19] DB05829 (1-34) , but not [ P55008 ,R19] DB05829 (1-28) , increased differentiation of Wt9 cells during a 16-day alternate-daily treatment protocol . Wt9 cells expressed PKC-betaI , -delta , -epsilon and -zeta , none of which exhibited net translocation to membranes in response to DB05829 (1-34) or either analog . DB05829 (1-34) induced activation of membrane-associated PKC-delta , however , and a time- and concentration-dependent increase in cytosolic [phospho-Thr505]PKC-delta which was maximal within 40 s at 100 nM in both Wt9 cells and primary osteoblasts . This response was mimicked by [ P55008 ,R19] DB05829 (1-34) but not by [ P55008 ,R19] DB05829 (1-28) . Increased expression of bone sialoprotein ( BSP ) and osteocalcin ( OC ) mRNAs induced by PTH(1-34) and [ P55008 ,R19] DB05829 (1-34) in Wt9 cells was blocked by rottlerin , a PKC-delta inhibitor . We conclude that PTH1Rs activate PKC-delta by a P98160 -independent , PTH(29-34)-dependent mechanism that promotes osteoblastic differentiation . P10275 -mediated antagonism of estrogen-dependent low density lipoprotein receptor transcription in cultured hepatocytes . Postmenopausal women receiving hormone replacement therapy have a lower risk of coronary heart disease than women who do not receive hormone treatment . Multiple mechanisms are likely to underlie estrogen 's cardioprotective action , including lowering of plasma low density lipoprotein ( LDL ) cholesterol . Using an in vitro system exhibiting normal regulation of P01130 ( P01130 ) gene transcription , we show that 17beta-estradiol activates the P01130 promoter in transiently transfected HepG2 cells . P01130 activation by estrogen in HepG2 cells is dependent on the presence of exogenous estrogen receptor , and the estrogen-responsive region of the P01130 promoter colocalizes with the sterol response element previously identified . The estrogen response is concentration dependent , saturable , and sensitive to antagonism by estrogen receptor antagonists . Further , we show that compounds with androgen receptor agonist activity attenuate the estrogen-induced up-regulation of P01130 in our model system . Progestins with androgen receptor agonist activity , such as medroxyprogesterone acetate , also suppress estrogen 's effects on P01130 expression through their androgenic properties . Characterization of the interplay between these hormone receptors on the P01130 in vitro system may allow a better understanding of the actions of sex steroids on P01130 gene expression and their roles in cardiovascular disease . 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 . Regulation of P14061 and P18405 in lymphocytes . We previously reported lymphocyte expression of genes encoding enzymes required for steroid metabolism ; however , only 17beta-HSD and 5alpha-reductase showed significant enzyme activity . We now investigate regulation of lymphocyte expression for genes encoding 17beta-HSD and 5alpha-reductase . Cultured human T and B lymphoid cell lines and peripheral blood mononuclear cells were treated with known regulators of steroidogenic gene expression including forskolin , PMA , ionomycin , various steroids , interleukin ( IL ) -4 , and P05231 . Treatment with 10 or 50 microM forskolin resulted in a 20-60 % reduction of expression for P14061 ( encoding 17beta-HSD I ) in T and B lymphoid cell lines and peripheral blood mononuclear cells , although such a change was not observed in the expression of P18405 ( encoding 5alpha-reductase I ) . No significant changes were found when cells were treated for 24 h with various concentrations of PMA or ionomycin . Incubation with 10(-9) to 10(-7) M androstenedione or estradiol increased expression of P14061 , while testosterone decreased the expression of this gene . P18405 expression was increased in the presence of 5alpha- DB02901 although no consistent changes were observed when the cells were treated with testosterone . Other steroids , including dexamethasone , progesterone , and 6-hydroxypregnanolone , produced no effects on expression of either P14061 or P18405 . Treatment with 0.1-10 ng/ml of P05112 or P05231 also did not effect significant changes in gene expression . These data implicate the involvement of the DB02527 -protein kinase signal transduction pathway in regulating lymphocyte expression of P14061 . Furthermore , it appears that lymphocyte P14061 and P18405 are regulated to some extent by specific steroids . P10275 and nutrient signaling pathways coordinate the demand for increased amino acid transport during prostate cancer progression . L-Type amino acid transporters such as Q01650 and O75387 mediate the uptake of essential amino acids . Here , we report that prostate cancer cells coordinate the expression of Q01650 and O75387 to maintain sufficient levels of leucine needed for mTORC1 signaling and cell growth . Inhibiting O43561 function was sufficient to decrease cell growth and mTORC1 signaling in prostate cancer cells . These cells maintained levels of amino acid influx through androgen receptor-mediated regulation of O75387 expression and P18848 regulation of Q01650 expression after amino acid deprivation . These responses remained intact in primary prostate cancer , as indicated by high levels of O75387 in primary disease , and by increased levels of Q01650 after hormone ablation and in metastatic lesions . Taken together , our results show how prostate cancer cells respond to demands for increased essential amino acids by coordinately activating amino acid transporter pathways vital for tumor outgrowth . [ Antibodies to the glutamate receptor ] . Glutamate receptors ( GluRs ) are classified into metabotropic GluRs and ionotropic GluRs . Ionotropic GluRs include the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ( AMPA ) -type , kainate ( KA ) -type , and N-methyl-D-aspartate ( DB01221 ) -type GluRs ( NRs ) . Antibodies to the NRs have been detected using immunoblot , cell-based assays , and enzyme linked immunosorbent assay ( ELISA ) . In patient with non-paraneoplastic , non-herpetic acute limbic encephalitis ( NHALE ) , antibodies against Q13224 ( GluRε2 ) and Q05586 ( Gluζ1 ) are detected in the sera and P04141 . In addition to the antibodies to NRs detected by ELISA , antibodies to an NR-complex detected by cell-based assay are found , not only in P04141 from NHALE , but also in P04141 from epilepsy , Creutzfeldt-Jakob disease ( CJD ) , etc . Antibodies to NRs internalize mainly extra-synaptic NRs , and dissociate the connection between Q12879 and P52799 receptor at the synapse . IgG fractions containing antibodies to NRs decrease apoptosis of cultured neurons . Antibodies to AMPA-type GluRs have been detected by immunoblot , cell-based assay and ELISA . In Rasmussen syndrome , antibodies against P42263 ( GluR3 ) were found to be the primary pathological factor . However the antibodies did not cause Rasmussen syndrome in mice models . These antibodies have been shown to cause excitotoxicity through P42263 , complement-dependent cytotoxicity , etc . Antibodies to P42261 / P42262 in paraneoplastic limbic encephalitis modulate expression and localization of P42261 / P42262 at the synapse . Antibodies to metabotropic GluRs have been detected using cell-based assays in patients with Hodgkin 's lymphoma . Passive transfer of the IgG fraction from patients having antibodies to metabotropic GluR1 causes ataxia in mice . The association between oxytocin receptor gene ( P30559 ) polymorphisms and affective temperaments , as measured by TEMPS-A . BACKGROUND : DB00107 is associated with social interaction , trust , and affectivity . Affective temperaments are traits based on Kraepelin 's typological definition of the " fundamental states " of manic-depressive illness . These states can be measured by the Temperament Evaluation of Memphis , Pisa , Paris and San Diego-Autoquestionnaire version ( TEMPS-A ) . The objective of this study is to assess the association between oxytocin receptor gene ( P30559 ) polymorphisms and affective temperaments . METHODS : Participants consisted of 493 genetically unrelated , non-clinical Japanese subjects ( 307 males and 186 females ) . The Mini-International Neuropsychiatric Interview ( MINI ) was used to screen and exclude those who had a lifetime diagnosis of schizophrenia or other psychotic disorders . Fifteen P30559 tag single nucleotide polymorphisms ( SNPs ) were genotyped using TaqMan® or direct sequencing . The Haploview 4.1. software determined the haplotype block structure . Haplotype-based quantitative trait association analysis with Bonferroni correction using PLINK 1.06 software was used to assess the association between haplotypes and the following affective temperaments : depressive , cyclothymic , hyperthymic , irritable , and anxious . RESULTS : Two haplotype blocks were identified on the P30559 . The depressive temperament was significantly associated with the most frequent haplotype GGGTGTC ( rs11131149/rs2243370/rs2243369/rs13316193/rs2254298/rs2268493/rs2268491 ) ( corrected P < 0.05 ) . LIMITATIONS : This study consisted of participants from a corporation and the effect sizes were small . CONCLUSIONS : The findings suggest that an P30559 haplotype is associated with a discrete depressive temperament . Clarification of the biological basis of this temperamental trait may help to elucidate the pathophysiology of depressive disorder . [ 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 . 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 . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . 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 . 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 . [ Production of granulocyte colony-stimulating factor and parathyroid hormone-related protein in squamous cell carcinoma of the renal pelvis accompanied with inferior vena cava thrombus ] . A 54-year-old man with general fatigue and lumbago was admitted for further examination of hypercalcemia and leukocytosis . CT showed a huge renal tumor and extension of the tumor thrombus to the inferior vena cava ( IVC ) . Moreover , the serum granulocyte colony-stimulating factor ( DB00099 ) and the C-terminal of parathyroid hormone-related protein ( P12272 ) were elevated . Under the diagnosis of advanced renal tumor , we performed nephro-ureterectomy and throbectomy . Pathological examination revealed squamous cell carcinoma of the renal pelvis . To our knowledge , this is the first case in Japan that of the simultaneous production of G- P04141 and P12272 in squamous cell carcinoma of the renal pelvis accompanied with IVC thrombus . 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 . Dysregulation of leukocyte gene expression in women with medication-refractory depression versus healthy non-depressed controls . BACKGROUND : Depressive Disorders ( DD ) are a great financial and social burden . Females display 70 % higher rate of depression than males and more than 30 % of these patients do not respond to conventional medications . Thus medication-refractory female patients are a large , under-served , group where new biological targets for intervention are greatly needed . METHODS : We used real-time quantitative polymerase chain reaction ( qPCR ) to evaluate mRNA gene expression from peripheral blood leukocytes for 27 genes , including immune , Q9Y251 -axis , ion channels , and growth and transcription factors . Our sample included 23 females with medication refractory DD : 13 with major depressive disorder ( MDD ) , 10 with bipolar disorder ( BPD ) . Our comparison group was 19 healthy , non-depressed female controls . We examined differences in mRNA expression in DD vs. controls , in MDD vs. BPD , and in patients with greater vs. lesser depression severity . RESULTS : DD patients showed increased expression for P22301 , P05231 , P30559 , Q99572 , P47900 , and Q8NER1 . BPD patients showed increased P05067 , P16220 , P19838 , P04150 , and P09486 and decreased P01375 expression . Depression severity was related to increased P22301 , P47900 , P51575 , and Q9HBA0 expression . CONCLUSIONS : These results support prior findings of dysregulation in immune genes , and provide preliminary evidence of dysregulation in purinergic and other ion channels in females with medication-refractory depression , and in transcription and growth factors in those with BPD . If replicated in future research examining protein levels as well as mRNA , these pathways could potentially be used to explore biological mechanisms of depression and to develop new drug targets . 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 . 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 . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . Translational PK-PD modelling of molecular target modulation for the AMPA receptor positive allosteric modulator Org 26576 . INTRODUCTION : The α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ( AMPA ) receptor potentiator Org 26576 represents an interesting pharmacological tool to evaluate the utility of glutamatergic enhancement towards the treatment of psychiatric disorders . In this study , a rat-human translational pharmacokinetic-pharmacodynamic ( PK-PD ) model of AMPA receptor modulation was used to predict human target engagement and inform dose selection in efficacy clinical trials . METHODS : Modelling and simulation was applied to rat plasma and cerebrospinal fluid ( P04141 ) pharmacokinetic and pharmacodynamic measurements to identify a target concentration ( EC(80) ) for AMPA receptor modulation . Human plasma pharmacokinetics was determined from 33 healthy volunteers and eight major depressive disorder patients . From four out of these eight patients , P04141 PK was also determined . Simulations of human P04141 levels were performed for several doses of Org 26576 . RESULTS : Org 26576 ( 0.1-10 mg/kg , i.v. ) potentiated rat hippocampal AMPA receptor responses in an exposure-dependant manner . The rat plasma and P04141 PK data were fitted by one-compartment model each . The rat P04141 PK-PD model yielded an EC(80) value of 593 ng/ml ( 90 % confidence interval 406.8 , 1,264.1 ) . The human plasma and P04141 PK data were simultaneously well described by a two-compartment model . Simulations showed that in humans at 100 mg QD , P04141 levels of Org 26576 would exceed the EC(80) target concentration for about 2 h and that 400 mg P55957 would engage AMPA receptors for 24 h . CONCLUSION : The modelling approach provided useful insight on the likely human dose-molecular target engagement relationship for Org 26576 . Based on the current analysis , 100 and 400 mg P55957 would be suitable to provide ' phasic ' and ' continuous ' AMPA receptor engagement , respectively . Phosphorylation of thymidylate synthase from various sources by human protein kinase CK2 and its catalytic subunits . P04818 ( TS ) was found to be a substrate for both catalytic subunits of human CK2 , with phosphorylation by CK2alpha and CK2alpha ' characterized by similar K(m) values , 4.6microM and 4.2microM , respectively , but different efficiencies , the apparent turnover number with CK2alpha being 10-fold higher . With both catalytic subunits , phosphorylation of human TS , like calmodulin and P55957 , was strongly inhibited in the presence of the regulatory subunit CK2beta , the holoenzyme being activated by polylysine . Phosphorylation of recombinant human , rat , mouse and Trichinella spiralis TSs proteins was compared , with the human enzyme being apparently a much better substrate than the others . Following hydrolysis and TLC , phosphoserine was detected in human and rat , and phosphotyrosine in T. spiralis , TS , used as substrates for CK2alpha . MALDI-TOF MS analysis led to identification of phosphorylated DB00133 (124) in human TS , within a sequence LGFS(124)TREEGD , atypical for a CK2 substrate recognition site . The phosphorylation site is located in a region considered important for the catalytic mechanism or regulation of human TS , corresponding to the loop 107-128 . Following phosphorylation by CK2alpha , resulting in incorporation of 0.4mol of phosphate per mol of dimeric TS , human TS exhibits unaltered K(m) values for DB03800 and N(5,10)-methylenetetrahydrofolate , but a 50 % lower turnover number , pointing to a strong influence of DB00133 (124) phosphorylation on its catalytic efficiency .

MH_train_1445

MH_train_1445

MH_train_1445

interacts_with DB00819?

multiple_choice

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 . Identification and characterization of an aquaporin 1 immunoreactive amacrine-type cell of the mouse retina . Using immunocytochemistry , a type of amacrine cell that is immunoreactive for aquaporin 1 was identified in the mouse retina . P29972 immunoreactivity was found in photoreceptor cells of the outer nuclear layer ( ONL ) and in a distinct type of amacrine cells of the inner nuclear layer ( INL ) . P29972 -immunoreactive ( IR ) amacrine cell somata were located in the INL and their processes extended through strata 3 and 4 of the inner plexiform layer ( Q53GA4 ) with thin varicosities . The density of the P29972 -IR amacrine cells increased from 100/mm(2) in the peripheral retina to 350/mm(2) in the central retina . The P29972 -IR amacrine cells comprise 0.5 % of the total amacrine cells . The P29972 -IR amacrine cell bodies formed a regular mosaic , which suggested that they represent a single type of amacrine cell . Double labeling with P29972 and glycine , gamma-aminobutyric acid ( GABA ) or Q99259 (65) antiserum demonstrated that the P29972 -IR amacrine cells expressed GABA or Q99259 (65) but not glycine . Their synaptic input was primarily from other amacrine cell processes . They also received synaptic inputs from a few cone bipolar cells . The primary synaptic targets were ganglion cells , followed by other amacrine cells and cone bipolar cells . In addition , gap junctions between an P29972 -IR amacrine process and another amacrine process were rarely observed . In summary , a GABAergic amacrine cell type labeled by an antibody against P29972 was identified in the mouse retina and was found to play a possible role in transferring a certain type of visual information from other amacrine or a few cone bipolar cells primarily to ganglion cells . 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 . Q07869 gamma 2 regulates adipose expression of the phosphoenolpyruvate carboxykinase gene . DB01819 carboxykinase ( PEPCK ) is expressed at high levels in liver , kidney , and adipose tissue . This enzyme catalyzes the rate-limiting step in hepatic and renal gluconeogenesis and adipose glyceroneogenesis . The regulatory factors important for adipose expression of the PEPCK gene are not well defined . Previous studies with transgenic mice established that the region between bp -2086 and -888 is required for expression in adipose tissue but not for expression in liver or kidney tissue . We show here that a DNA fragment containing this region can function as an enhancer and direct differentiation-dependent expression of a chloramphenicol acetyltransferase gene from a heterologous promoter in cultured 3T3-F442A preadipocytes and adipocytes . We further demonstrate that the adipocyte-specific transcription factor Q07869 gamma 2 , previously identified as a regulator of the adipocyte P2 enhancer , binds in a heterodimeric complex with RXR alpha to the PEPCK 5'-flanking region at two sites , termed P35558 ( bp -451 to -439 ) and Q16822 ( bp -999 to -987 ) . Forced expression of Q07869 gamma 2 and RXR alpha activates the PEPCK enhancer in non-adipose cells . This activation is potentiated by peroxisome proliferators and fatty acids but not by 9-cis retinoic acid . Mutation of the Q07869 gamma 2 binding site ( Q16822 ) abolishes both the activity of the enhancer in adipocytes and its ability to be activated by Q07869 gamma 2 and RXR alpha . These results establish a role for Q07869 gamma 2 in the adipose expression of the PEPCK gene and suggest that this factor functions as a coordinate regulator of multiple adipocyte-specific genes . Development of a cell-based assay for the detection of anti-aquaporin 1 antibodies in neuromyelitis optica spectrum disorders . OBJECTIVE : To develop a cell-based assay ( CBA ) to detect aquaporin 1 ( P29972 ) antibodies and determine sensitivity/specificity in patients with neuromyelitis optica ( NMO ) spectrum disorders . METHODS : A P29320 -293T transfected cell model expressing P29972 was established and detected to be serum P29972 antibodies . RESULTS : P29972 antibodies were present in 73/98 ( 74.5 % ) P55087 antibody-positive patients . Some P55087 antibody-negative patients were also P29972 antibody-positive . Test sensitivity was 74.5 % in 98 P55087 antibody-positive patients . Test specificity was 79.6 % in 67 multiple sclerosis ( MS ) patients and 31 controls . CONCLUSION : A sensitive and simple CBA was developed to detect serum P29972 antibodies . P29972 antibodies were mainly present in NMO and its high-risk syndrome , but also in some MS patients . Taurine biosynthesis in rat brain : a new specific and sensitive microassay of cysteine sulfinate decarboxylase ( CSDI ) activity through selective immunotrapping and its use for distribution studies . DB00151 sulfinate decarboxylase ( Q9Y600 ) , the putative biosynthetic enzyme for taurine , has been shown to exist in two forms in rat brain , respectively CSDI and CSDII , one of which ( CSDII ) is considered to be in fact glutamate decarboxylase ( Q99259 ) . CSDI assay after immunotrapping was made possible by using an anti- Q9Y600 antiserum raised in sheep immunized with a partially purified Q9Y600 fraction from liver . This antiserum immunoprecipitated both liver Q9Y600 and brain CSDI activities with the same affinity but did not inhibit their enzymatic activities . The immunotrapping of CSDI was selective without any contamination by Q99259 /CSDII activity . The immunotrapped Q9Y600 activity , which corresponded exactly to the amount of Q9Y600 not precipitated by a Q99259 /CSDII antiserum , was not inhibited by a specific irreversible Q99259 inhibitor . A quantitative , selective and sensitive assay was thus developed by measuring Q9Y600 activity on the solid phase after immunotrapping . Kinetic parameters of the immunotrapped enzyme remained unchanged . CSDI activity represented only a fraction , around 20 % with saturating concentration of substrate , of the total Q9Y600 activity in rat brain homogenate . This indicates that most studies on total Q9Y600 activity dealt essentially with CSDII activity that is indeed Q99259 . Regional and subcellular distributions of CSDI have been determined . CSDI activity was about threefold higher in the richest ( cerebellum ) compared to the poorest ( striatum ) region without any correlation with Q99259 /CSDII distribution . Subcellular distribution showed a fourfold enrichment of CSDI activity in the synaptosomal fraction . The precise role of CSDI and CSDII in the biosynthesis of taurine in vivo remains to be elucidated . 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 . Hyperosmolar mannitol simulates expression of aquaporins 4 and 9 through a p38 mitogen-activated protein kinase-dependent pathway in rat astrocytes . The membrane pore proteins , aquaporins ( AQPs ) , facilitate the osmotically driven passage of water and , in some instances , small solutes . Under hyperosmotic conditions , the expression of some AQPs changes , and some studies have shown that the expression of P29972 and P55064 is regulated by MAPKs . However , the mechanisms regulating the expression of P55087 and AQP9 induced by hyperosmotic stress are poorly understood . In this study , we observed that hyperosmotic stress induced by mannitol increased the expression of P55087 and AQP9 in cultured rat astrocytes , and intraperitoneal infusion of mannitol increased P55087 and AQP9 in the rat brain cortex . In addition , a p38 MAPK inhibitor , but not P29323 and JNK inhibitors , suppressed their expression in cultured astrocytes . AQPs play important roles in maintaining brain homeostasis . The expression of P55087 and AQP9 in astrocytes changes after brain ischemia or traumatic injury , and some studies have shown that p38 MAPK in astrocytes is activated under similar conditions . Since mannitol is commonly used to reduce brain edema , understanding the regulation of AQPs and p38 MAPK in astrocytes under hyperosmotic conditions induced with mannitol may lead to a control of water movements and a new treatment for brain edema . [ The chemical structure and pharmacological properties of a novel isoxazolidinedione insulin sensitizer , DB04971 ] . DB04971 is an isoxazolidine-3,5-dione derivative . This drug activates both Q07869 gamma and Q07869 alpha , and shows not only a hypoglycemic effect but also a stronger triglyceride-lowering effect than the thiazolidine-2,4-diones . DB04971 improved both the impaired insulin-stimulated autophosphorylation levels of Zucker fatty rats and impaired insulin-induced P14672 translocation to the plasma membrane as well as insulin-induced glucose uptake in high fat diet rats , indicating that DB04971 enhances insulin signaling and reduces insulin resistance . Furthermore , DB04971 prevented several diabetic complications , such as cataract , nephropathy , and neuropathy in Zucker diabetic fatty rats . As a non-thiazolidinedione insulin sensitizer , DB04971 has been the first to start clinical trials and is currently undergoing evaluation in clinical studies for diabetic patients . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . 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 . 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 . 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 . Aquaporin 1-positive stromal niche-like cells directly interact with P19022 -positive clusters in the basal limbal epithelium . Stem cells have a specialized microenvironment for maintaining self-renewal and multipotent capacities . It is believed that a cornea epithelial stem cell niche exists in the limbus . To characterize the niche of limbal epithelial stem cells , we observed the limbal basal epithelial layer by histological analysis . Cell clusters or cell suspensions from limbal tissue were prepared with collagenase or dispase II and fixed for cytospin sections . Adhesion assays were done to quantitate calcium-dependent cell adhesion . Limbal tissue and cytospin sections were analyzed by immunohistochemistry , transmission electron microscopy and confocal microscopy . P29972 positive ( P29972 (+) ) cells were observed as non-epithelial cells in the subepithelial stroma . P29972 expression did not co-localize with CD31 , podoplanin , Q16655 positive cells , but were observed in vimentin positive stromal cells . When we made a thorough search of limbal basal cells by confocal microscopy , P29972 (+) were observed in the proximity of N-cad , P19012 and p63 positive limbal basal epithelial cells . Furthermore , electron microscope revealed stromal cells penetrating the epithelial basal membrane and forming calcium-dependent cellular adhesions with N-cad(+) limbal basal epithelial cells . Although we could not clearly detect the expression of N-cad in the P29972 (+) cells , P29972 (+) cells immediately beneath the epithelial basement membrane may be stromal niche-like cells that directly interact with N-cad(+) limbal basal epithelial progenitor cells . 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 . 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 . Combined Effects of Q07869 γ Agonists and Epidermal Growth Factor Receptor Inhibitors in Human Proximal Tubule Cells . We aimed to determine whether epidermal growth factor receptor ( P00533 ) inhibition , in addition to a peroxisome proliferator-activated receptor gamma ( Q07869 γ ) agonist , prevents high-glucose-induced proximal tubular fibrosis , inflammation , and sodium and water retention in human proximal tubule cells exposed to normal glucose ; high glucose ; high glucose with the Q07869 γ agonist pioglitazone or with the P- P00533 inhibitor , gefitinib ; or high glucose with both pioglitazone and gefitinib . We have shown that high glucose increases AP-1 and NF κ B binding activity , downstream phosphorylation of P00533 and Erk1/2 , and fibronectin and collagen IV expression . Pioglitazone reversed these effects but upregulated P48764 and P29972 expression . Gefitinib inhibited high glucose induced fibronectin and collagen IV , and P00533 and Erk1/2 phosphorylation and reversed pioglitazone-induced increases in P48764 and P29972 expression . Our data suggests that combination of an P00533 inhibitor and a Q07869 γ agonist mitigates high-glucose-induced fibrosis and inflammation and reverses the upregulation of transporters and channels involved in sodium and water retention in human proximal tubule cells . Hence P00533 blockade may hold promise , not only in limiting tubulointerstitial pathology in diabetic nephropathy , but also in limiting the sodium and water retention observed in patients with diabetes and exacerbated by Q07869 γ agonists . 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 . Agonist-induced glycogenolysis in rabbit retinal slices and cultures . 1. The effects of different putative retinal transmitters and/or modulators on glycogenolysis in rabbit retinal slices and in retinal Müller cell cultures were examined . 2 . Incubation of rabbit retinal slices or primary retinal cultures ( either 3-5 day-old or 25-30 day-old ) in a buffer solution containing [ 3H ] -glucose resulted in the accumulation of newly synthesized [ 3H ] -glycogen . 3 . DB00368 ( NA ) , isoprenaline , vasoactive intestinal peptide ( P01282 ) , 5-hydroxytryptamine ( 5-HT ) and 8-hydroxy-dipropylaminetetralin ( 8-OH-DPAT ) stimulated the hydrolysis of this newly formed 3H-polymer . The potency order of maximal stimulations was : P01282 greater than NA greater than isoprenaline greater than 5-HT greater than 8-OH-DPAT . 4 . The putative retinal transmitters , dopamine , gamma-aminobutyric acid ( GABA ) , glycine and taurine and the muscarinic agonist carbachol ( CCh ) had no effect on [ 3H ] -glycogen content . 5 . The glycogenolytic effects of NA/isoprenaline and 5-HT/8-OH-DPAT appear to be mediated by beta-adrenoceptors and 5-HT1 receptors ( possibly P08908 ) , respectively while the P01282 -induced response involved another receptor subtype . 6 . Agonists which mediated [ 3H ] -glycogen hydrolysis also stimulated an increase in adenosine 3':5'-cyclic monophosphate ( cyclic AMP ) formation . Both responses are blocked to a similar extent by the same antagonists and so are probably mediated via the same receptor subtypes . Moreover , dibutyryl cyclic AMP ( db cyclic AMP ) promoted tritiated glycogen breakdown in the three retinal preparations . 7 . Not all receptors linked to cyclic AMP production however promote glycogenolysis . Dopamine and apomorphine stimulated cyclic AMP formation via D1-receptors without influencing glycogenolysis . These receptors are exclusively associated with neurones . P20142 -1α encoded by the Q9UBK2 gene regulates oxidative energy metabolism in equine skeletal muscle during exercise . Peroxisome proliferator-activated receptor-γ coactivator 1α ( P20142 -1α ) has emerged as a critical control factor in skeletal muscle adaptation to exercise , acting via transcriptional control of genes responsible for angiogenesis , fatty acid oxidation , oxidative phosphorylation , mitochondrial biogenesis and muscle fibre type composition . In a previous study , we demonstrated a significant increase in mRNA expression for the gene encoding P20142 -1α ( Q9UBK2 ) in Thoroughbred horse skeletal muscle following a single bout of endurance exercise . In this study , we investigated mRNA expression changes in genes encoding transcriptional coactivators of P20142 -1α and genes that function upstream and downstream of P20142 -1α in known canonical pathways . We used linear regression to determine the associations between Q9UBK2 mRNA expression and expression of the selected panel of genes . Biopsy samples were obtained from the gluteus medius pre-exercise ( T(0) ) , immediately post-exercise ( T(1) ) and 4 h post-exercise ( T(2) ) . Significant ( P < 0.05 ) expression fold change differences relative to T(0) were detected for genes functioning in angiogenesis ( ANGP2 and P15692 ) ; Ca(2+) -dependent signalling pathway ( Q08209 ) ; carbohydrate/glucose metabolism ( Q16654 ) ; fatty acid metabolism/mitochondrial biogenesis ( PPPARGC1B ) ; haem biosynthetic process ( P13196 ) ; insulin signalling ( Q12778 , PPPARGC1A and P14672 ) ; mitogen-activated protein kinase signalling ( Q16539 and Q02078 ) ; and myogenesis ( Q9UKV0 ) . Gene expression associations were identified between Q9UBK2 and genes involved in angiogenesis , mitochondrial respiration , glucose transport , insulin signalling and transcriptional regulation . These results suggest that P20142 -1α and genes regulated by P20142 -1α play significant roles in the skeletal muscle response to exercise and therefore may contribute to performance potential in Thoroughbred horses .

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interacts_with DB08879?

multiple_choice

Comparison of the P00533 resistance mutation profiles generated by P00533 -targeted tyrosine kinase inhibitors and the impact of drug combinations . Recent clinical data indicates that the emergence of mutant drug-resistant kinase alleles may be particularly relevant for targeted kinase inhibitors . In order to explore how different classes of targeted therapies impact upon resistance mutations , we performed P00533 ( epidermal-growth-factor receptor ) resistance mutation screens with erlotinib , lapatinib and DB05424 . Distinct mutation spectra were generated with each inhibitor and were reflective of their respective mechanisms of action . DB01259 yielded the widest variety of mutations , whereas mutational variability was lower in the erlotinib and DB05424 screens . DB01259 was uniquely sensitive to mutations of residues located deep within the selectivity pocket , whereas mutation of either DB00145 (796) or DB00151 (797) resulted in a dramatic loss of DB05424 potency . The clinically observed T790M mutation was common to all inhibitors , but occurred with varying frequencies . Importantly , the presence of C797S with T790M in the same P00533 allele conferred complete resistance to erlotinib , lapatinib and DB05424 . The combination of erlotinib and DB05424 effectively reduced the number of drug-resistant clones , suggesting a possible clinical strategy to overcome drug resistance . Interestingly , our results also indicate that co-expression of ErbB2 ( v-erb-b2 erythroblastic leukaemia viral oncogene homologue 2 ) has an impact upon the P00533 resistance mutations obtained , suggesting that ErbB2 may play an active role in the acquisition of drug-resistant mutations . Mass spectrometry-based quantification of myocardial protein adducts with acrolein in an in vivo model of oxidative stress . Acrolein ( P10323 ) exposure leads to the formation of protein- P10323 adducts . Protein modification by P10323 has been associated with various chronic diseases including cardiovascular and neurodegenerative diseases . Here , we report an analytical strategy that enables the quantification of Michael-type protein adducts of P10323 in mitochondrial proteome samples using liquid chromatography in combination with tandem mass spectrometry and selected ion monitoring ( LC-MS/MS P19623 ) analysis . Our approach combines site-specific identification and relative quantification at the peptide level of protein- P10323 adducts in relation to the unmodified protein thiol pool . Treatment of 3-month-old rats with CCl(4) , an established in vivo model of acute oxidative stress , resulted in significant increases in the ratios of distinct P10323 -adducted peptides to the corresponding unmodified thiol-peptides obtained from proteins that were isolated from cardiac mitochondria . The mitochondrial proteins that were found adducted by P10323 were malate dehydrogenase , DB00157 dehydrogenase [ ubiquinone ] flavoprotein 1 , cytochrome c oxidase subunit VIb isoform 1 , DB00171 synthase d chain , and P12235 . The findings indicate that protein modification by P10323 has potential value as an index of mitochondrial oxidative stress . 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 . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Atheroprotective effect of oleoylethanolamide ( OEA ) targeting oxidized LDL . Dietary fat-derived lipid oleoylethanolamide ( OEA ) has shown to modulate lipid metabolism through a peroxisome proliferator-activated receptor-alpha ( Q07869 -α ) -mediated mechanism . In our study , we further demonstrated that OEA , as an atheroprotective agent , modulated the atherosclerotic plaques development . In vitro studies showed that OEA antagonized oxidized LDL ( ox-LDL ) -induced vascular endothelial cell proliferation and vascular smooth muscle cell migration , and suppressed lipopolysaccharide ( LPS ) -induced LDL modification and inflammation . In vivo studies , atherosclerosis animals were established using balloon-aortic denudation ( Q92934 ) rats and ApoE(-/-) mice fed with high-caloric diet ( HCD ) for 17 or 14 weeks respectively , and atherosclerotic plaques were evaluated by oil red staining . The administration of OEA ( 5 mg/kg/day , intraperitoneal injection , i.p. ) prevented or attenuated the formation of atherosclerotic plaques in HCD- Q92934 rats or HCD-ApoE(-/-) mice . Gene expression analysis of vessel tissues from these animals showed that OEA induced the mRNA expressions of Q07869 -α and downregulated the expression of M-CFS , an atherosclerotic marker , and genes involved in oxidation and inflammation , including P35228 , P35354 , P01375 -α and P05231 . Collectively , our results suggested that OEA exerted a pharmacological effect on modulating atherosclerotic plaque formation through the inhibition of LDL modification in vascular system and therefore be a potential candidate for anti-atherosclerosis drug . A cyclooxygenase-2/prostaglandin E2 pathway augments activation-induced cytosine deaminase expression within replicating human B cells . Within inflammatory environments , B cells encountering foreign or self-Ag can develop tertiary lymphoid tissue expressing activation-induced cytosine deaminase ( Q9GZX7 ) . Recently , this DNA-modifying enzyme was detected in nonlymphoid cells within several inflamed tissues and strongly implicated in malignant transformation . This study examines whether a cyclooxygenase 2 ( P35354 ) pathway , often linked to inflammation , influences Q9GZX7 expression in activated B lymphocytes . In this paper , we report that dividing human B cells responding to surrogate C3d-coated Ag , P05112 , and Q9Y275 express Q9GZX7 , as well as P35354 . A progressive increase in Q9GZX7 with each division was paralleled by a division-related increase in a P35354 -linked enzyme , microsomal PGE(2) synthase-1 , and the PGE(2)R , EP2 . Cells with the greatest expression of Q9GZX7 expressed the highest levels of EP2 . Although P35354 inhibitors diminished both Q9GZX7 expression and IgG class switching , exogenous PGE(2) and butaprost , a selective EP2 agonist , augmented Q9GZX7 mRNA/protein and increased the numbers of IgG(+) progeny . Despite the latter , the proportion of IgG(+) cells within viable progeny generally declined with PGE(2) supplementation . This was not due to PGE(2)-promoted differentiation to plasma cells or to greater downstream switching . Rather , because phosphorylated ataxia telangiectasia mutated levels were increased in progeny of PGE(2)-supplemented cultures , it appears more likely that PGE(2) facilitates Q9GZX7 -dependent DNA double-strand breaks that block B cell cycle progression or promote activation-induced cell death , or both . Taken together , the results suggest that a PGE(2) feed-forward mechanism for augmenting P35354 pathway proteins promotes progressively increased levels of Q9GZX7 mRNA , protein , and function . MicroRNA-19b functions as potential anti-thrombotic protector in patients with unstable angina by targeting tissue factor . The activation of a hemostatic system plays a critical role in the incidence of acute coronary events . Hemostatic proteins may be regulated by microRNAs ( miRNAs ) . Microparticles ( MPs ) are the major carrier of circulating miRNAs . The aim of this study was to determine the potential role of miRNAs in regulating gene expression involved in the hemostatic system in patients with unstable angina ( UA ) . MiRNA expression profiles in the plasma from patients with UA ( UA group , n=9 ) compared with individuals with clinical suspicion of coronary artery disease ( CAD ) but negative angiography ( control group , n=9 ) showed that among 36 differentially expressed miRNAs , miR-19b was the most obvious one . Using real-time PCR , 5 selected miRNA levels in plasma ( UA group , n=20 ; control group , n=30 ) and plasma MPs ( UA group n=6 ; control group n=6 ) were proved to be consistent with the miRNA array . Flow cytometry analysis indicated that the amounts of plasma endothelial microparticles ( EMPs ) were increased in UA patients ( UA group , n=4 ) compared to controls ( control group , n=4 ) . In cultured endothelial cells ( ECs ) , P01375 -α increased miR-19b release and expression . P13726 ( TF ) was predicted to be the target of miR-19b by bioinformatics analysis . Luciferase reporter assays demonstrated that miR-19b binds to TF mRNA . Overexpression of miR-19b inhibited TF expression and procoagulant activity . This study indicates that in UA patients , the increase of miR-19b wrapped in EMPs due to endothelial dysfunction may partially contribute to the circulating miR-19b elevation and miR-19b may play an anti-thrombotic role by inhibiting the expression of TF in ECs . Islet transplantation in type 1 diabetes mellitus using cultured islets and steroid-free immunosuppression : Miami experience . Following the success obtained with transplantation of fresh human islets under steroid-free immunosuppression , this trial evaluated the transplantation of islets that had undergone a period of in vitro culture and the potential of tumor necrosis factor ( P01375 ) blockade to improve islet engraftment . Subjects included 16 patients with type 1 diabetes mellitus ( T1DM ) ; half were randomly assigned to receive DB00065 immediately preceding initial infusion . Immunosuppression consisted of daclizumab induction and sirolimus/tacrolimus maintenance . Out of 16 subjects 14 achieved insulin independence with one or two islet infusions ; adverse events precluded completion in two . Without supplemental infusions , 11/14 ( 79 % ) subjects were insulin independent at 1 year , 6/14 ( 43 % ) at 18 months ; these same subjects remain insulin independent at 33+/-6 months . While on immunosuppression , all patients maintained graft function . Out of 14 patients , 8 suffered chronic partial graft loss , likely immunological in nature , 5 of these received supplemental infusions . Currently , 11 subjects remain on immunosuppression , 8 ( 73 % ) are insulin independent , two with supplemental infusions . P01308 independent subjects demonstrated normalization of HbA1c , fructosamine and Mean Amplitude of Glycemic Excursions ( MAGE ) values . No clinical benefit of infliximab was identified . These results demonstrate that transplantation of cultured human islet allografts results in reproducible insulin independence in all subjects under this immunosuppressive regimen , comparable to that of freshly transplanted islets ( Edmonton protocol ) . 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 . Coordinate suppression of B cell lymphoma by P60484 and Q92835 phosphatases . The inositol phosphatases phosphatase and tensin homologue ( P60484 ) and Src homology 2 domain-containing inositol phosphatase ( Q92835 ) negatively regulate phosphatidylinositol-3-kinase ( PI3K ) -mediated growth , survival , and proliferation of hematopoietic cells . Although deletion of P60484 in mouse T cells results in lethal T cell lymphomas , we find that animals lacking P60484 or Q92835 in B cells show no evidence of malignancy . However , concomitant deletion of P60484 and Q92835 ( bPTEN/ Q92835 (-/-) ) results in spontaneous and lethal mature B cell neoplasms consistent with marginal zone lymphoma or , less frequently , follicular or centroblastic lymphoma . bPTEN/ Q92835 (-/-) B cells exhibit enhanced survival and express more Q07820 and less Bim . These cells also express low amounts of p27(kip1) and high amounts of cyclin D3 and thus appear poised to undergo proliferative expansion . Unlike normal B cells , bPTEN/ Q92835 (-/-) B cells proliferate to the prosurvival factor B cell activating factor ( Q9Y275 ) . Interestingly , although Q9Y275 availability may promote lymphoma progression , we demonstrate that Q9Y275 is not required for the expansion of transferred bPTEN/ Q92835 (-/-) B cells . This study reveals that P60484 and Q92835 act cooperatively to suppress B cell lymphoma and provides the first direct evidence that Q92835 is a tumor suppressor . As such , assessment of both P60484 and Q92835 function are relevant to understanding the etiology of human B cell malignancies that exhibit augmented activation of the PI3K pathway . Analysis of actin microfilaments and cell-to-substrate adhesive structures in human fibroblasts from individuals genetically predisposed to colonic carcinoma . Skin fibroblasts from patients with inherited adenomatosis of the large bowel ( P10323 -SF ) possess alterations in actin microfilament ( MF ) organization which serve to distinguish " predisposed " cells from fibroblasts derived from normal individuals ( P46459 ) . MF bundle frequency and diameter were considerably reduced in P10323 -SF compared to P46459 . This deficit in MF density correlated with a 60 % decline in cytoskeletal-associated actin half-life . Absence of a well-structured MF network in P10323 -SF was reflected in relatively poor cell-to-substrate adhesion ( as indicated by increased sensitivity to trypsin release ) and extensive membrane ruffling . Unlike P46459 , P10323 -SF failed to develop well-defined vinculin-containing focal contacts although the cellular content of vinculin was approximately the same in both cell types . The relatively low substrate adhesivity and reduced incidence of adhesive structures ( i.e. , MF and associated focal contacts ) which typify P10323 -SF correlated with a sixfold increase in cellular plasminogen activator ( PA ) activity . This increased protease activity corresponded with a 50-70 % reduction in the content of the PA inhibitor-like protein p50 in both the saponin-resistant undersurface matrix and the culture medium . Increased motility and reduced cell-to-substrate adhesion , involving several cellular structural elements , appear to be significant correlates of the " predisposed " phenotype in cultured fibroblasts . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Beyond inflammation : airway epithelial cells are at the interface of innate and adaptive immunity . It has become increasingly clear that airway epithelial cells are central participants in innate and adaptive immune responses as well as mucosal inflammation . Epithelial cells produce antimicrobial host defense molecules , proinflammatory cytokines and chemokines in response to activation via pathogen recognition receptors . Recruitment of immune cells including dendritic cells , T cells and B cells into the proximity of epithelium results in the enhancement of adaptive immunity through interactions with epithelial cells . Newly identified epithelial-derived cytokines , including Q969D9 , O95760 and Q9Y275 , help to shape the local accumulation and activation of Th2 responses and B cell immunoglobulin production . Epithelial cells are also downstream targets of molecules that activate IL-13R and P00533 and are responsible for mucus production in both protective immune responses and allergic airway inflammatory diseases . Improved understanding of epithelial immune and inflammatory responses will hopefully suggest new strategies for therapeutic intervention . 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 . The effects of nepafenac and amfenac on retinal angiogenesis . PURPOSE : DB06802 is a potent NSAID that rapidly penetrates the eye following topical ocular administration . In the eye , nepafenac is converted to amfenac , which has unique time-dependent inhibitory properties for P23219 and P35354 . The purpose of the present study was to investigate the capacity of amfenac to inhibit discrete aspects of the angiogenic cascade in vitro , and to test the efficacy of amfenac and nepafenac in vivo , using the rat OIR model . METHODS : Müller cells were treated with amfenac , celecoxib ( P35354 ) , or SC-560 ( P23219 ) , and hypoxia-induced P15692 and PGE(2) assessed . Endothelial cells were treated with amfenac , celecoxib , or SC-560 , and P15692 -induced proliferation and tube formation assessed . Rat pups were subjected to OIR , received intravitreal injections of amfenac , celecoxib , or SC-560 , and neovascularization ( NV ) , prostanoid production , and P15692 assessed . Other OIR-exposed pups were treated with topical nepafenac , ketorolac , or diclofenac , and inhibition of NV assessed . RESULTS : Amfenac treatment failed to inhibit hypoxia-induced P15692 production . Amfenac treatment significantly inhibited P15692 -induced tube formation and proliferation by EC . Amfenac treatment significantly reduced retinal prostanoid production and NV in OIR . DB06802 treatment significantly reduced retinal NV in OIR ; ketorolac and diclofenac had no effect . CONCLUSIONS : DB06802 and amfenac inhibit OIR more effectively than the commercially available topical and injectable NSAIDs used in this study . Our data suggests there are P36551 -dependent and P36551 -independent mechanisms by which amfenac inhibits OIR . Because it is bioavailable to the posterior segment following topical delivery , nepafenac appears to be a promising advancement in the development of therapies for neovascular eye diseases . 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 . DB02709 activates AMPK and suppresses LPS-induced NF-κB-dependent P35354 activation in RAW 264.7 macrophage cells . AMP-activated protein kinase ( AMPK ) , an enzyme involved in energy homeostasis , regulates inflammatory responses , but its precise mechanisms are not fully understood . Recent evidence has shown that resveratrol ( RES ) , an AMPK activator , reduces prostaglandin E(2) production in lipopolysaccharide ( LPS ) -treated microglia . Here , we examined the effect of RES on nuclear factor kappa B ( NF-κB ) dependent cyclooxygenase ( P36551 ) -2 activation in LPS-treated RWA 264.7 macrophages . We found that treatment with RES increased AMPK activation . AMPK and acetyl DB01992 carboxylase phosphorylation were attenuated in cells treated with LPS+RES , compared to cells treated with LPS alone . RES inhibited tumor necrosis factor ( P01375 ) -α and P01375 receptor 1 in LPS-treated cells . Finally , RES inhibited LPS-induced NF-κB translocation into the nucleus and P35354 expression . Moreover , the effects of 5-aminoimidazole-4-carboxamide ribose and compound C were consistent with the effects of RES in LPS-treated cells . Taken together , these results suggest that the anti-inflammatory action of RES in RAW 264.7 macrophages is dependent on AMPK activation and is associated with inhibition of the LPS-stimulated NF-κB-dependent P35354 signaling pathway . P11926 is a critical determinant of P04198 oncogenesis and a therapeutic target in neuroblastoma . Neuroblastoma is a frequently lethal childhood tumor in which MYC gene deregulation , commonly as P04198 amplification , portends poor outcome . Identifying the requisite biopathways downstream of MYC may provide therapeutic opportunities . We used transcriptome analyses to show that P04198 -amplified neuroblastomas have coordinately deregulated myriad polyamine enzymes ( including P11926 , P19623 , P52788 , P17707 , O95190 , and Q9NWM0 ) to enhance polyamine biosynthesis . High-risk tumors without P04198 amplification also overexpress P11926 , the rate-limiting enzyme in polyamine biosynthesis , when compared with lower-risk tumors , suggesting that this pathway may be pivotal . Indeed , elevated P11926 ( independent of P04198 amplification ) was associated with reduced survival in a large independent neuroblastoma cohort . As polyamines are essential for cell survival and linked to cancer progression , we studied polyamine antagonism to test for metabolic dependence on this pathway in neuroblastoma . The Odc inhibitor alpha-difluoromethylornithine ( DB06243 ) inhibited neuroblast proliferation in vitro and suppressed oncogenesis in vivo . DB06243 treatment of neuroblastoma-prone genetically engineered mice ( TH- P04198 ) extended tumor latency and survival in homozygous mice and prevented oncogenesis in hemizygous mice . In the latter , transient Odc ablation permanently prevented tumor onset consistent with a time-limited window for embryonal tumor initiation . Importantly , we show that DB06243 augments antitumor efficacy of conventional cytotoxics in vivo . This work implicates polyamine biosynthesis as an arbiter of P04198 oncogenesis and shows initial efficacy for polyamine depletion strategies in neuroblastoma , a strategy that may have utility for this and other MYC-driven embryonal tumors .

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Critical role of P21453 and integrin β4 in P14210 /c- DB00134 -mediated increases in vascular integrity . Vascular endothelial cell ( EC ) barrier integrity is critical to vessel homeostasis whereas barrier dysfunction is a key feature of inflammatory disorders and tumor angiogenesis . We previously reported that hepatocyte growth factor ( P14210 ) -mediated increases in EC barrier integrity are signaled through a dynamic complex present in lipid rafts involving its receptor , c- DB00134 . We extended these observations to confirm that P21453 ( sphingosine 1-phosphate receptor 1 ) and integrin β4 ( P16144 ) are essential participants in P14210 -induced EC barrier enhancement . Immunoprecipitation experiments demonstrated P14210 -mediated recruitment of c- DB00134 , P16144 and P21453 to caveolin-enriched lipid rafts in human lung EC with direct interactions of c- DB00134 with both P21453 and P16144 accompanied by c- DB00134 -dependent P21453 and P16144 transactivation . Reduced P21453 expression ( siRNA ) attenuated both P16144 and Rac1 activation as well as c- DB00134 / P16144 interaction and resulted in decreased transendothelial electrical resistance . Furthermore , reduced P16144 expression attenuated P14210 -induced c- DB00134 activation , c- DB00134 / P21453 interaction , and effected decreases in Q14703 - and P14210 -induced EC barrier enhancement . Finally , the c- DB00134 inhibitor , DB05030 , suppressed P14210 -induced c- DB00134 activation as well as P21453 and P16144 transactivation . These results support a critical role for P21453 and P16144 transactivation as rate-limiting events in the transduction of P14210 signals via a dynamic c- DB00134 complex resulting in enhanced EC barrier integrity . P04035 inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells . OBJECTIVE : Q9UEF7 is thought to play a critical role in the development of age-related disorders including arteriosclerosis . Statins may exert vascular protective effects , independent of the lowering of plasma cholesterol levels . We investigated the impact of statins on mRNA expression of the age-suppressor gene , klotho in mIMCD3 cells . METHODS AND RESULTS : Q9UEF7 mRNA levels were evaluated with real-time RT-PCR . DB01076 and pitavastatin increased the expression of klotho mRNA in a dose-dependent manner . This stimulatory effect was abolished by the addition of mevalonate , GGPP and FPP , essential molecules for isoprenylation of the small GTPase Rho . As was the case with the statin treatment , inhibition of Rho-kinase by Y27632 up-regulated klotho mRNA . In contrast to the statin treatment , stimulation with angiotensin II down-regulated klotho mRNA expression without obvious morphological changes . Furthermore , pretreatment with atorvastatin blunted the angiotensin II-induced response and ameliorated the decrease in klotho mRNA expression towards basal levels . RhoA activity was further evaluated by detection of its translocation . Angiotensin II activated RhoA , whereas statins potently inactivated RhoA and blocked RhoA activation by angiotensin II . CONCLUSION : Statins inactivate the RhoA pathway , resulting in over-expression of klotho mRNA , which may contribute to the novel pleiotropic effects of statins towards vascular protection . DB01076 ameliorates tissue damage of obstructed ureter in rats . AIMS : To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitor on the tissue damage and fibrosis of obstructed ureters , 80 rats were studied . MAIN METHODS : DB01076 , a P04035 inhibitor , was administered to 40 rats at the dose of 20 mg/kg per day 1day before unilateral ligation of ureters and every day thereafter . The other rats served as controls . Eight rats from each group were sacrificed for examination on days 7 , 14 , 21 , 28 and 42 after ligation , respectively . The expressions of transforming growth factor-β1 ( TGF-β1 ) , Interleukine-1β ( IL-1β ) , Interleukine-6 ( P05231 ) , tumor necrosis factor-alpha ( P01375 -α ) , proliferation cell nuclear antigen ( P12004 ) , and the apoptotic cells in the ureteric smooth muscle were examined . KEY FINDINGS : Hydroureter and fibrosis of the muscle layer became progressively aggravated in the ligated ureters of the atorvastatin-treated group and control group . The severities of hydroureter and muscle layer fibrosis in the ligated ureters of the treated group were significantly less than in the control group . The atorvastatin administration also decreased the expression of TGF-β1 , IL-1β , P05231 , P01375 -α , P12004 and the labeling index of apoptotic cells in the smooth muscle layer of ligated ureters in the treated group . SIGNIFICANCE : We concluded that atorvastatin might ameliorate the tissue damage of obstructed ureters , at least partially , via the inhibition on TGF-β1 ) expression and by diminishing the effects of pro-inflammatory cytokines . Perturbation of beta1-integrin function alters the development of murine mammary gland . The expression of a transgene coding for a chimeric molecule , containing the cytoplasmic and transmembrane domains of the beta1-integrin chain and the extracellular domain of the T-cell differentiation antigen P01730 , was targeted to the mouse mammary gland by the mouse mammary tumor virus ( MMTV ) promoter . The chimera does not interact with the extracellular ligands ; however , its expression in cultured cells was shown to interfere with focal adhesion kinase ( Q05397 ) phosphorylation following ligation of endogenous beta1-integrin . Therefore , expression of the transgenic protein on the cell surface should uncouple adhesion from intracellular events associated with the beta1-cytoplasmic domain and thus perturb beta1-integrin functions . Although most of the transgenic females were able to lactate , their mammary glands had a phenotype clearly distinct from that of wild-type mice . At mid-pregnancy and the beginning of lactation , transgenic glands were underdeveloped and the epithelial cell proliferation rates were decreased , while the apoptosis levels were higher than in wild-type glands . In lactation , the amounts of the whey acidic protein ( WAP ) and beta-casein gene transcripts were diminished , and the basement membrane component , laminin and the beta4-integrin chain accumulated at the lateral surface of luminal epithelial cells , revealing defects in polarization . Our observations prove that in vivo , beta1-integrins are involved in control of proliferation , apoptosis , differentiation and maintenance of baso-apical polarity of mammary epithelial cells , and therefore are essential for normal mammary gland development and function . Q9UEF7 has dual protective effects on cisplatin-induced acute kidney injury . Q9UEF7 protects the kidney from ischemia-reperfusion injury , but its effect on nephrotoxins is unknown . Here we determined whether Q9UEF7 protects the kidney from cisplatin toxicity . DB00515 increased plasma creatinine and induced tubular injury , which were exaggerated in Q9UEF7 haplosufficient ( Kl/+ ) and ameliorated in transgenic Q9UEF7 overexpressing ( Tg-Kl ) mice . P80188 and active caspase-3 protein and the number of apoptotic cells in the kidney were higher in Kl/+ and lower in Tg-Kl compared with wild-type mice . Q9UEF7 suppressed basolateral uptake of cisplatin by the normal rat kidney cell line ( Q7Z2Y5 ) , an effect similar to cimetidine , a known inhibitor of organic cation transport ( O75051 ) . A decrease in cell surface and total OCT2 protein and O75051 activity by Q9UEF7 was mimicked by β-glucuronidase . The Q9UEF7 effect was attenuated by β-glucuronidase inhibition . On the other hand , OCT2 mRNA was reduced by Q9UEF7 but not by β-glucuronidase . Moreover , cimetidine inhibited O75051 activity but not OCT2 expression . Unlike cimetidine , Q9UEF7 reduced cisplatin-induced apoptosis from either the basolateral or apical side and even when added after Q7Z2Y5 cells were already loaded with cisplatin . Thus , Q9UEF7 protects the kidney against cisplatin nephrotoxicity by reduction of basolateral uptake of cisplatin by OCT2 and a direct anti-apoptotic effect independent of cisplatin uptake . Q9UEF7 may be a useful agent to prevent and treat cisplatin-induced nephrotoxicity . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . 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 . 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 . DB06589 synergizes with docetaxel in the treatment of bladder cancer cells . OBJECTIVES : To investigate the efficacy of pazopanib , both alone and in combination with docetaxel , in bladder cancer cells . Bladder cancer expresses many potential therapeutic targets of biological agents , including the vascular endothelial growth factor receptor ( VEGFR ) . DB06589 is a small molecule inhibitor of P17948 , -2-3 , platelet-derived growth factor receptor ( P09619 ) , and c-Kit . MATERIALS AND METHODS : Using human bladder cancer cells HTB3 , HT1376 , J82 , RT4 , CRL1749 , T24 , Sup , and HTB9 , the treatment effect of pazopanib and cytotoxic chemotherapy was assessed using a tetrazolium-based assay . The combinatorial effect of these agents on clonogenic growth was further examined . Western blotting was used to assess changes in relevant downstream targets , including phospho-AKT , phospho- Q05397 , total AKT , and total Q05397 . RESULTS : Single-agent pazopanib had modest activity . However , synergy was seen with the combination of docetaxel and pazopanib in these multiple cells lines . J82 and T24 cells were selected for additional clonogenic testing because of their resistance to single-agent docetaxel chemotherapy . 1.25 nM of docetaxel had little effect on clonogenic formation ; however , in combination with pazopanib , significant inhibition of colony formation was observed . This combination treatment additionally decreased phospho-AKT , an important mediator of cell survival in all cell lines , whereas phospho- Q05397 expression was variably affected . CONCLUSIONS : The present study demonstrates synergistic efficacy of pazopanib with docetaxel in docetaxel-resistant bladder cancer cells . This work supports future evaluation of pazopanib with docetaxel for the treatment of bladder cancer with the potential of improved efficacy and toxicity . 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 . 5-hydroxytryptamine and its receptors in systemic vascular walls . 5-hydroxytryptamine ( 5-HT ) in the bloodstream is largely contained in platelets and circulates throughout the entire vascular system . 5-HT released from activated platelets dramatically changes the function of vascular smooth muscle cells ( VSMCs ) and endothelial cells ( ECs ) . In VSMCs , 5-HT induces proliferation and migration via 5- Q13049 receptors . These effects are further enhanced by vasoactive substances such as thromboxane A2 and angiotensin II . 5- Q13049 receptor activation in VSMCs also causes both enhancement of prostaglandin I2 production by inducing cyclooxygenase-2 and reduction of nitric oxide ( NO ) by suppressing inducible NO synthase . Evidence showing that 5-HT in ECs plays a principal role in angiogenesis now exists . Stimulation of 5-HT1 and/or 5-HT2 receptors has been implicated in the angiogenic effect of 5-HT . The extracellular signal-regulated kinase and endothelial NO synthase ( P29474 ) activation-dependent pathways are involved in the mechanisms . Moreover , Q13639 receptors in ECs have been shown to also regulate angiogenesis . Recent reports show sarpogrelate , a selective antagonist of the 5- Q13049 receptor , indirectly enhances the function of P28222 receptors in ECs via inhibition of 5- Q13049 receptors in VSMCs or platelets . This indirect action of P28222 receptors in ECs may increase NO production derived from P29474 and a vasodilator response . Furthermore , sarpogrelate and other 5- Q13049 receptor antagonists have been shown to reduce the constitutive activity of 5- Q13049 receptors . It is believed that increasing evidence on the role of 5-HT receptors will contribute to the expansion of the clinical application of existing therapeutic drugs such as sarpogrelate , and to the development of new 5-HT receptor-related drugs for treating cardiovascular diseases . The immunosuppressant FTY720 prolongs survival in a mouse model of diet-induced coronary atherosclerosis and myocardial infarction . FTY720 , an analogue of sphingosine-1-phosphate , is cardioprotective during acute injury . Whether long-term FTY720 affords cardioprotection is unknown . Here , we report the effects of oral FTY720 on ischemia/reperfusion injury and in hypomorphic apoE mice deficient in Q8WTV0 receptor expression ( ApoeR61(h/h)/ Q8WTV0 ( -/- mice ) , a model of diet-induced coronary atherosclerosis and heart failure . We added FTY720 ( 0.3 mg·kg(-1)·d(-1) ) to the drinking water of C57BL/6J mice . After ex vivo cardiac ischemia/reperfusion injury , these mice had significantly improved left ventricular ( LV ) developed pressure and reduced infarct size compared with controls . Subsequently , ApoeR61(h/h)/ Q8WTV0 (-/-) mice fed a high-fat diet for 4 weeks were treated or not with oral FTY720 ( 0.05 mg·kg(-1)·d(-1) ) . This sharply reduced mortality ( P < 0.02 ) and resulted in better LV function and less LV remodeling compared with controls without reducing hypercholesterolemia and atherosclerosis . Oral FTY720 reduced the number of blood lymphocytes and increased the percentage of P01730 +Foxp3+ regulatory T cells ( Tregs ) in the circulation , spleen , and lymph nodes . FTY720-treated mice exhibited increased TGF-β and reduced IFN-γ expression in the heart . Also , P01730 expression was increased and strongly correlated with molecules involved in natural Treg activity , such as TGF-β and Q9Y5U5 . Our data suggest that long-term FTY720 treatment enhances LV function and increases longevity in mice with heart failure . These benefits resulted not from atheroprotection but from systemic immunosuppression and a moderate reduction of inflammation in the heart . The role of atorvastatin in regulating the immune response leading to vascular damage in a model of Kawasaki disease . Superantigens have been implicated in a number of diseases including Kawasaki disease ( KD ) , a multi-system vasculitis resulting in coronary artery aneurysms . We have characterized a murine disease model in which coronary arteritis is induced by a novel superantigen found in Lactobacillus casei cell wall extract ( LCWE ) . Using this animal model of KD , we have identified three pathogenic steps leading to coronary artery aneurysm formation . These steps include T cell activation and proliferation , production of the proinflammatory cytokine tumour necrosis factor ( P01375 ) -α and up-regulation of matrix metalloproteinase 9 ( P14780 ) , an elastolytic protease . In addition to their cholesterol-lowering effects , 3-hydroxy-3-methylglutaryl ( HMG ) coenzyme A ( DB01992 ) reductase inhibitors ( statins ) have pleotropic immunomodulatory properties . Thus , we examined the effect of atorvastatin in modulating each of these three critical pathogenic processes leading to aneurysm formation in the disease model . DB01076 inhibited lymphocyte proliferation in response to superantigen stimulation in a dose-dependent manner . This inhibition was also observed for production of soluble mediators of inflammation including interleukin ( IL ) -2 and P01375 -α . The inhibitory effect on proliferation was rescued completely by mevalonic acid , confirming that the mechanism responsible for this inhibitory activity on immune activation was inhibition of P04035 . Similarly , P01375 -α-induced P14780 production was reduced in a dose-dependent manner in response to atorvastatin . Inhibition of extracellular-regulated kinase ( P29323 ) phosphorylation appears to be the mechanism responsible for inhibition of P14780 production . In conclusion , atorvastatin is able to inhibit critical steps known to be important in the development of coronary aneurysms , suggesting that statins may have therapeutic benefit in patients with KD . The heme oxygenase system rescues hepatic deterioration in the condition of obesity co-morbid with type-2 diabetes . The prevalence of non-alcoholic fatty-liver disease ( NAFLD ) is increasing globally . NAFLD is a spectrum of related liver diseases that progressive from simple steatosis to serious complications like cirrhosis . The major pathophysiological driving of NAFLD includes elevated hepatic adiposity , increased hepatic triglycerides/cholesterol , excessive hepatic inflammation , and hepatocyte ballooning injury is a common histo-pathological denominator . Although heme-oxygenase ( HO ) is cytoprotective , its effects on hepatocyte ballooning injury have not been reported . We investigated the effects of upregulating HO with hemin or inhibiting it with stannous-mesoporphyrin ( DB04912 ) on hepatocyte ballooning injury , hepatic adiposity and inflammation in Zucker-diabetic-fatty rats ( ZDFs ) , an obese type-2-diabetic model . DB03404 administration to ZDFs abated hepatic/plasma triglycerides and cholesterol , and suppressed several pro-inflammatory cytokines and chemokines including , P01375 -α , P05231 , IL-1β , macrophage-inflammatory-protein-1α ( MIP-1α ) and macrophage-chemoattractant-protein-1 ( P13500 ) , with corresponding reduction of the pro-inflammatory M1-phenotype marker , Q92838 and hepatic macrophage infiltration . Correspondingly , hemin concomitantly potentiated the protein expression of several markers of the anti-inflammatory macrophage-M2-phenotype including ED2 , P22301 and CD-206 , alongside components of the HO-system including P09601 , HO-activity and cGMP , whereas the HO-inhibitor , DB04912 abolished the effects . Furthermore , hemin attenuated liver histo-pathological lesions like hepatocyte ballooning injury and fibrosis , and reduced extracellular-matrix/profibrotic proteins implicated in liver injury such as osteopontin , TGF-β1 , fibronectin and collagen-IV . We conclude that hemin restore hepatic morphology by abating hepatic adiposity , suppressing macrophage infiltration , inflammation and fibrosis . The selective enhancement of anti-inflammatory macrophage-M2-phenotype with parallel reduction of pro-inflammatory macrophage-M1-phenotype and related chemokines/cytokines like P01375 -α , P05231 , IL-1β , MIP-1α and P13500 are among the multifaceted mechanisms by which hemin restore hepatic morphology . DB08860 , an P04035 inhibitor , exerts P29474 -independent protective actions against angiotensin II induced cardiovascular remodeling and renal insufficiency . Angiotensin II ( Ang II ) plays a pivotal role in cardiovascular remodeling leading to hypertension , myocardial infarction , and stroke . DB08860 , an P04035 inihibitor , is known to have pleiotropic actions against the development of cardiovascular remodeling . The objectives of this study were to clarify the beneficial effects as well as the mechanism of action of pitavastatin against Ang II-induced organ damage . C57BL6/J mice at 10 weeks of age were infused with Ang II for 2 weeks and were simultaneously administered pitavastatin or a vehicle . DB08860 treatment improved Ang II-induced left ventricular hypertrophy and diastolic dysfunction and attenuated enhancement of cardiac fibrosis , cardiomyocyte hypertrophy , coronary perivascular fibrosis , and medial thickening . Ang II-induced oxidative stress , cardiac TGFbeta-1 expression , and Smad 2/3 phosphorylation were all attenuated by pitavastatin treatment . DB08860 also reduced Ang II-induced cardiac remodeling and diastolic dysfunction in P29474 -/- mice as in wild-type mice . In P29474 -/- mice , the Ang II-induced cardiac oxidative stress and TGF-beta-Smad 2/3 signaling pathway were enhanced , and pitavastatin treatment attenuated the enhanced oxidative stress and the signaling pathway . Moreover , pitavastatin treatment reduced the high mortality rate and improved renal insufficiency in Ang II-treated P29474 -/- mice , with suppression of glomerular oxidative stress and TGF-beta-Smad 2/3 signaling pathway . In conclusion , pitavastatin exerts P29474 -independent protective actions against Ang II-induced cardiovascular remodeling and renal insufficiency through inhibition of the TGF-beta-Smad 2/3 signaling pathway by suppression of oxidative stress . P04035 inhibitor , atorvastatin , promotes sensorimotor recovery , suppressing acute inflammatory reaction after experimental intracerebral hemorrhage . BACKGROUND AND PURPOSE : Statins have neuroprotective effects on ischemic stroke . They modify the endothelial function , increase blood flow , and inhibit thrombus formation , which are independent of lipid-lowering effects . However , whether statins have a protective effect toward hemorrhagic stroke is yet unknown . To test this possibility , we attempted to determine the effect of atorvastatin on experimental intracerebral hemorrhage ( ICH ) . METHODS : ICH was induced using stereotaxic infusion of collagenase into the left basal ganglia in adult rats . DB01076 ( 1 mg/kg or 10 mg/kg ) or phosphate-buffered saline was administered for 2 weeks . To monitor the sensorimotor deficits , limb placing and Rotorod tests were performed . Hematoma volume , brain water content , and hemispheric atrophy were analyzed . Immunohistochemical staining for myeloperoxidase ( P05164 ) , microglia ( OX42 ) , inducible nitric oxide synthase ( P35228 ) , or endothelial nitric oxide synthase ( P29474 ) was performed . Perihematomal cell death was determined by TUNEL staining . RESULTS : The atorvastatin-treated ICH group showed better performance on Rotorod and limb placing tests when compared with the vehicle-treated group ( P < 0.01 ) . The hematoma volumes between groups were not different , but the brain water content and hemispheric atrophy were reduced in the atorvastatin-treated ICH group . DB01076 reduced TUNEL-positive cells , P35228 expression , and P05164 -positive or OX42-positive cells in the perihematomal regions in a dose-dependent manner , whereas it increased P29474 expression . CONCLUSIONS : The present study shows that atorvastatin reduces the perihematomal cell death via antiinflammation , which is associated with sensorimotor recovery after experimental ICH . 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 . DB01076 induces insulin sensitization in Zucker lean and fatty rats . BACKGROUND : The 3-hydroxy-3-methyl glutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors ( ' statins ' ) have been implicated in preventing new onset type 2 diabetes , whereas the mechanism of this effect is not known . We investigated the effects of an P04035 inhibitor , atorvastatin , on insulin sensitization in Zucker lean and fatty rats . METHODS AND RESULTS : In vivo studies of insulin sensitization were performed in chow fed Zucker lean and fatty rats treated with atorvastatin 50mg/kg/day ( ATORVA_50 ) and results were compared to Zucker lean and fatty rats treated with drug vehicle only ( CONT ) . Additional Zucker lean rats were treated with an intermediate dose of atorvastatin 25mg/kg/day ( ATORVA_25 ) . Treatment with atorvastatin resulted in a dose-dependent improvement in whole body insulin sensitivity in both lean and fatty rats , with an approximately two-fold increase in glucose infusion rate and glucose disposal ( Rd ) in ATORVA_50 versus CONT ( p < 0.01 ) . DB01076 50mg/kg/day resulted in an increase in DB08831 ( 2-DOG ) uptake by skeletal muscles ( approximately two-fold increase in 2-DOG uptake in quadriceps ( p=0.06 ) and gastrocnemius ( p < 0.01 ) ) in lean Zucker rats . P01308 -stimulated phosphorylation of Akt/ P31749 was significantly increased in skeletal muscle of ATORVA_50 versus CONT in both lean and fatty rats . CONCLUSION : DB01076 induces insulin sensitization in Zucker lean and fatty rats . This may be a clinically important pleiotropic effect if confirmed in insulin resistant humans . 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 . Investigation of immunomodulatory properties of human Wharton 's Jelly-derived DB05914 after lentiviral transduction . Human Wharton 's Jelly-derived Mesenchymal Stem Cells ( hWJ-MSCs ) are considered as an alternative for bone-marrow-derived MSCs . These cells have immunosuppressive properties . It was unclear whether the WJ-MSCs would sustain their immunomodulatory characteristics after lentiviral transduction or not . In this study , we evaluated immunomodulatory properties of WJ-MSCs after lentiviral transduction . HWJ-MSCs were transduced with lentiviral particles . Expression of transduced and un-transduced hWJ-MSCs surface molecules and secretion of P22301 , P14210 , P15692 and TGF-β was analyzed . Cell proliferation and frequency of P01730 (+)CD25(+) CD127(low/neg) Foxp3(+) T regulatory cells was measured . There was no difference between the surface markers and secretion of P22301 , P14210 , P15692 and TGF-β in transduced and un-transduced hWJ-MSCs . Both cells inhibited the proliferation of PHA stimulated PBMCs , and improved the frequency of T regulatory cells . These findings suggest that lentiviral transduction does not alter the immunomodulatory function of hWJ-MSCs . However , lentiviral transduction may have a wide range of applications in gene therapy .

MH_train_1448

MH_train_1448

MH_train_1448

interacts_with DB00784?

multiple_choice

Impact of follicle stimulating hormone receptor variants in fertility . PURPOSE OF REVIEW : Genetic variation plays a crucial role in modification of normal or disease pathophysiology . Follicle stimulating hormone receptor ( P23945 ) signaling is necessary for normal development and function of the ovaries and testes . Here , we review the associations between P23945 polymorphisms and fertility or subfertility . RECENT FINDINGS : P23945 polymorphisms consist of single nucleotide changes within the coding and regulatory regions and/or alternatively spliced products . Most of the investigations focused on two single nucleotide polymorphisms ( SNPs ) in the coding region of the receptor , which result in amino acid changes ( p.307Thr/Ala , p.680Asn/ DB00133 ) . In women , these SNPs were associated with variable response to ovarian stimulation with DB00094 during infertility treatment . Not all studies revealed an association , and those that did showed a small effect . Alternative splice variants of the receptor affecting the extracellular domain without causing a frameshift have been found in women undergoing ovarian stimulation , and in infertile men . Associations with polycystic ovary syndrome , premature ovarian failure , osteoporosis , and cancer found small effect . SUMMARY : The identification of P23945 variants in a select infertility patient population has significant clinical implications in demonstrating a possible genetic cause to female infertility and improves our understanding of the genetic basis of infertility as a whole . Pharmacogenomics is a new field aiming to devise individualized treatments for disorders based on the genetic signature of the patients . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . 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 . P35354 promotes early atherosclerotic lesion formation in P01130 -deficient mice . BACKGROUND : Atherosclerosis has features of an inflammatory disease . Because cyclooxygenase ( P36551 ) -2 is expressed in atherosclerotic lesions and promotes inflammation , we tested the hypotheses that selective P35354 inhibition would reduce early lesion formation in P01130 -deficient ( P01130 -/- ) mice and that macrophage P35354 expression contributes to atherogenesis in P01130 -/- mice . METHODS AND RESULTS : Treatment of male P01130 -/- mice fed the Western diet with rofecoxib or indomethacin for 6 weeks resulted in significant reductions in atherosclerosis in the proximal aorta ( 25 % and 37 % ) and in the aorta en face ( 58 % and 57 % ) , respectively . DB00533 treatment did not inhibit platelet thromboxane production , a P23219 -mediated process , but it significantly reduced the urinary prostacyclin metabolite 2,3-dinor-6-keto-PGF1alpha . Fetal liver cell transplantation was used to generate P01130 -/- mice null for expression of the P35354 gene by macrophages . After 8 weeks on the Western diet , P35354 -/- --> P01130 -/- mice developed significantly less ( 33 % to 39 % ) atherosclerosis than control P35354 +/+ --> P01130 -/- mice . In both the inhibitor studies and the transplant studies , serum lipids did not differ significantly between groups . CONCLUSIONS : The present studies provide strong pharmacological and genetic evidence that P35354 promotes early atherosclerotic lesion formation in P01130 -/- mice in vivo . These results support the potential of anti-inflammatory approaches to the prevention of atherosclerosis . Differentiation in vivo of classical non-steroidal antiinflammatory drugs from cytokine suppressive antiinflammatory drugs and other pharmacological classes using mouse tumour necrosis factor alpha production . The stimulation of tumour necrosis factor alpha ( P01375 alpha ) production by lipopolysaccharide ( LPS ) has been widely used , both in vitro and in vivo , to examine the biochemistry and pharmacology of inflammatory cytokine production . It appears that classical nonsteroidal antiinflammatory drugs ( NSAIDs ) ( prostaglandin H synthase 1 ( P23219 ) inhibitors ) do not inhibit but instead stimulate cytokine production . In the current study , the authors utilized LPS-induced P01375 alpha production in the Balb/c mouse to evaluate the activity of a classical NSAID , a mixed inhibitor , and SmithKline Beecham cytokine suppressive antiinflammatory drugs ( CSAID ) . The results corroborated the stimulation of P01375 alpha production by NSAIDs ( indomethacin , naproxen , ibuprofen ) and indicated that the stimulation rank-ordered with the potency of inhibition of P23219 . Neither acetaminophen nor nabumetone was found to stimulate P01375 alpha production significantly . Tenidap , a compound reported to inhibit P09917 , cyclooxygenase and cytokine production , also stimulated P01375 alpha production while the P09917 inhibitor , phenidone , was inactive . The CSAID ( exemplified by SK & F 86002 , SK & F 105809 and SK & F 104351 ) , strongly inhibited P01375 alpha production in this model system ( ED50s of 32 , 48 , and 34 mg/kg p.o. , respectively ) . These results clearly differentiate CSAID from the other compounds tested and suggest that CSAID are relatively weak inhibitors of PGHS 1 while being potent inhibitors of inflammatory cytokine production . 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 . Platelet activation in hypertension associated with hypercholesterolemia : effects of irbesartan . AIM : The aim of this study was to determine the effect of simultaneous hypertension and hypercholesterolemia on platelet activation , nitric oxide ( NO ) production and oxidative stress , and to evaluate the role of irbesartan , an angiotensin II type 1 receptor antagonist . METHODS : Golden Syrian hamsters were divided into three groups : controls , C ( fed a standard diet ) ; hypertensive-hypercholesterolemic , HH ( fed a diet enriched in 3 % cholesterol , 15 % butter and 8 % NaCl , for 4 months ) ; and hypertensive-hypercholesterolemic treated with irbesartan , HHI ( fed as HH group , plus irbesartan 10 mg kg(-1) per day , for 4 months ) . RESULTS : Compared with the C group , platelets isolated from the HH group showed : morphological modifications ; increased integrin β3 exposure and protein expression of P16109 , Q05397 , PI3K , Akt and Src ; reduced P29474 protein expression and NO production ; higher generation of ROS , mostly produced by NADPH-oxidase , cyclooxygenase-1 ( P23219 ) and 12-lipoxygenase ; and enhanced NAD(P)H oxidase activity and protein expression of gp91phox and P13498 subunits , 12-lipoxygenase , P23219 , cPLA(2) and PKC . Compared with the HH group , the treatment with irbesartan ( HHI group ) significantly attenuates the changes in all the molecules tested , reduces platelet aggregation , and improves intraplatelet redox balance . CONCLUSIONS : Experimental hypertension associated with hypercholesterolemia produces major changes in morphology , signaling mechanisms and oxidative stress in blood platelets . These changes were significantly diminished by irbesartan administration , which functions as an antioxidant on platelets . DB01370 uptake and effects on transferrin mediated iron uptake in primary cultures of rat neurons , astrocytes and oligodendrocytes . P02787 ( Tf ) is known primarily for its role in the transport and cellular uptake of iron ( Fe ) . Tf is also the major serum binding protein for Al . In this study , primary rat oligodendrocyte , neuron and astrocyte cultures were found to differ in Tf mediated Fe and Al uptake and in the effect of Al-Tf on Fe-Tf uptake during 4 h incubation periods . When incubated with Al-Tf ( 1.25 microM ) , oligodendrocytes displayed a 3- to 4-fold increase ( p=.0002 ) in Al , neurons demonstrated a much smaller ( p=.06 ) increase , and no increase was seen for astrocytes . When incubated with equimolar Al citrate or Al chloride , no increase in cellular Al was seen in any of the three cell types . Oligodendrocytes , astrocytes and neurons all demonstrated greater 59Fe uptake from Fe-Tf than Fe chloride . This uptake could be inhibited by excess Fe-Tf in oligodendrocytes and neurons , but not astrocytes . A small but significant inhibition of 59Fe uptake from Fe-Tf was seen after addition of Al-Tf to the incubation medium of oligodendrocytes , but not neurons or astrocytes . Oligodendrocytes may be particularly vulnerable to the accumulation of excess intracellular Al , and to interference of Al with Fe uptake . Such effects could contribute to Al-induced neurotoxicity if they result in altered myelin formation or maintenance . 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 . Topical glucocorticoids downregulate P23219 positive cells in nasal polyps . BACKGROUND : Influx of inflammatory cells is one of the hallmarks of nasal polyposis . As glucocorticoids ( GC ) are known to exhibit strong anti-inflammatory effects , these drugs are frequently used in the treatment of the disease . Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis . As cyclooxygenases ( P36551 ) are key enzymes in the synthesis of both pro- ( P23219 , P35354 ) and anti-inflammatory prostanoids ( P35354 ) , we investigated the role of topical GC on P23219 , P35354 and inflammatory markers in nasal polyps ( NP ) . METHODS : Immunohistochemical analysis of inflammatory markers ( P34810 , CD117 , MBP , elastase , IgE , BB-1 , P05112 , P05113 and P05231 ) , P23219 and P35354 was performed on normal nasal mucosa ( NM ) ( n = 18 ) , non-GC treated NP ( n = 27 ) and topical GC treated NP ( n = 12 ) . NP groups were matched for allergy , asthma and ASA intolerance . RESULTS : Increased numbers of eosinophils , P05113 + cells and IgE+ cells and decreased numbers of mastcells are striking features of NP inflammation ( P < 0.05 ) . In addition , increased numbers of P23219 + cells are observed in NP epithelium compared to NM ( P < 0.05 ) . CONCLUSION : Topical GC significantly reduce the number of P23219 + NP cells ( P < 0.05 ) , but have no significant effect on P35354 + NP cells . No significant reduction in the number of eosinophils is observed for GC treated NP . The number of P05113 + cells is however increased significantly upon GC treatment ( P < 0.05 ) . 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 . Alpha-tocopherol prevents cyclosporin A-mediated activation of phospholipase A2 and inhibition of Na+ , K(+)-adenosine triphosphatase activity in cultured hamster renal tubular cells . At concentrations of 0.5 microM and upward , cyclosporin A ( DB00091 ) caused dose-related inhibition of the growth of a hamster renal tubular cell line ( Q86TB3 ATCC ; Q16663 ) in vitro . Inhibition of cell growth was due to the cytotoxic properties of DB00091 which were associated with enhancement of activity of phospholipase A2 ( P04054 ) according to the increased generation of arachidonic acid and lysophosphatidylcholine ( Q16549 ) . Arachidonate per se , at concentrations of up to 20 microM , did not affect the growth of Q86TB3 cells , while cyclooxygenase and P09917 inhibitors failed to protect the cells against the antiproliferative effects of DB00091 . However , Q16549 caused dose-related inhibition of the growth of Q86TB3 cells . Moreover , coincubation with lysophospholipase or DB00163 ( AT , vitamin E ) , a P04054 inhibitory and lysophospholipid-complexing agent , protected the Q86TB3 cells against both DB00091 and Q16549 . The Na+ , K(+)-ATPase activity of Q86TB3 cells was also inhibited by DB00091 , with the enzyme being protected by inclusion of AT or lysophospholipase . Increased activity of P04054 and inhibition of Na+ , K(+)-ATPase preceded cytotoxicity and cytolysis . Excessive production of lysophospholipids and consequent inhibition of Na+ , K(+)-ATPase in renal tubular cells is a possible mechanism of DB00091 -induced nephrotoxicity . The protective effects of AT suggest that this agent may be clinically useful in preventing the renal side effects of DB00091 . Immune-related genes expression profile in orange-spotted grouper during exposure to Cryptocaryon irritans . Cryptocaryon irritans is one of the most important ectoparasites of marine fish . To identify the potential role of immune-related genes in antiparasitic immune responses in fish , we monitored the expression change of P10145 , P35354 , C-type lectin and transferrin in local and systemic immune organs of orange-spotted grouper post-C. irritans infection . P10145 expression was up-regulated during the course of infection in the skin , while P35354 and transferrin expression was up-regulated in the gill . P35354 expression was significantly down-regulated in the spleen ( 0·7-5 % of its control ) and head kidney ( 0·5-4 % of its control ) post-primary infection . P02787 expression was also down-regulated in the spleen and head kidney from 6 h to 5 days post-primary infection . However , C-type lectin expression was up-regulated in all tested organs post-infection , with the exception of day 7 in the spleen post-primary infection where the expression level was slightly down-regulated ( 44 % of its control ) . These results suggest that these four immune-related genes play an important role in grouper anti-C. irritans infection and that local immune organs as the active organs contribute more than systemic immune organs to this course . Combined adenine phosphoribosyltransferase and P34059 deficiency . We describe a Czech patient with combined adenine phosphoribosyltransferase ( P07741 ) deficiency ( 2,8-dihydroxyadenine urolithiasis ) and P34059 ( P34059 ) deficiency ( mucopolysaccharidosis Type IVA , Morquio disease A ) . DB00173 and its extremely insoluble derivative , 2,8-dihydroxyadenine , were identified in the urine , and P07741 deficiency was confirmed in erythrocytes . There was excessive excretion of keratan sulfate in the urine , and P34059 deficiency was confirmed in leukocytes . P34059 and P07741 are both located on chromosome 16q24.3 , suggesting that the patient had a deletion involving both genes . PCR amplification of genomic DNA indicated that a novel junction was created by the fusion of sequences distal to P34059 exon 2 and proximal to P07741 exon 3 , and that the size of the deleted region was approximately 100 kb . The deletion breakpoints were localized within P34059 intron 2 and P07741 intron 2 . Several other genes , including the alpha subunit of cytochrome B ( P13498 ) , which is deleted or mutated in the autosomal form of chronic granulomatous disease , are located in the 16q24.3 region , but PCR amplification showed that this gene was present in the proband . A patient with hemizygosity for P34059 deficiency and P07741 deficiency has been reported from Japan recently . These findings indicate that : ( i ) P07741 is located telomeric to P34059 ; ( ii ) P34059 and P07741 are transcribed in the same orientation ( centromeric to telomeric ) ; and ( iii ) combined P07741 / P34059 deficiency may be more common than hitherto realized . 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 . Increasing cell plating density mimics an early post-LH stage in cultured bovine granulosa cells . Cultured ovarian granulosa cells are essential models to study molecular mechanisms of gene regulation during folliculogenesis . Here , we characterize primary tissue culture models for bovine granulosa cells by morphological and physiological parameters and by novel molecular luteinization markers , as transcript abundance and DNA methylation levels . The data show that : ( 1 ) collagen substrate increased the number of attached , viable cells ; ( 2 ) the expression of the key transcripts of estrogen synthesis , P11511 , could be induced and maintained in granulosa cells from small to medium but not from large follicles , whereas ( 3 ) only granulosa cells from large but not from smaller follicles were responsive to LH ; ( 4 ) serum supplementation unfavorably transformed the cellular phenotype , induced proliferation and P12004 expression , reduced or abolished the transcript abundance of steroidogenic key genes and of gonadotropin receptor genes , P05108 , P11511 , P23945 and P22888 but , however , did not increase the abundance of the luteinization-specific marker transcripts P35354 , PTX3 , P41220 and O95498 ; but ( 5 ) by increasing the plating density , estradiol production and the abundance of P11511 transcripts , in particular those derived from the main ovarian promoter P2 , were decreased concurrently leaving P2-specific DNA methylation levels unchanged , whereas progesterone secretion was stimulated and the expression of both luteinization-specific marker transcripts , P41220 and O95498 , was significantly induced . From these data , we conclude that increasing the plating density induces a different , partly complementary , physiological and gene expression profile in cultured bovine granulosa cells and drives the cells towards an early post-LH stage of luteinization , even in the absence of luteinizing agents . Sulfonylurea and non-sulfonylurea hypoglycemic agents : pharmachological properties and tissue selectivity . DB00171 -sensitive K+ ( K( DB00171 ) ) channels play many important roles in cellular functions , including control of membrane excitability of skeletal muscle and neurons , K+ recycling in renal epithelia , cytoprotection in cardiac ischemia , and insulin secretion from pancreatic beta-cells . K( DB00171 ) channels are composed of pore-forming inwardly rectifying potassium channel ( Kir6.2 or Kir6.1 ) subunits and sulfonylurea receptor ( Q09428 , SUR2A , or SUR2B ) subunits . Kir6.2 or Kir6.1 subunits conjoined with a Q09428 subunit constitute the various tissue-specific K( DB00171 ) channels with distinct pharmacological properties . Both sulfonylureas and non-sulfonylurea hypoglycemic agents are used in treatment of type 2 diabetes mellitus . While the sulfonylurea receptor ( Q09428 ) is the target molecule of all of these hypoglycemic agents , the binding sites differ according to the moiety containing in the agent , and alter the pharmachological properties . In addition , chronic exposure of pancreatic beta-cells to the various agents affects the agent-specific sensitivities differently . Here we distinguish differences in pharmacological profile among the various hypoglycemic agents that reflect their chemical composition . We also suggest possible risk in the use of certain hypoglycemic agents in patients with ischemic heart disease . 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 . 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 . 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 .

MH_train_1449

MH_train_1449

MH_train_1449

interacts_with DB00477?

multiple_choice

Effect of gene disruptions of the TCA cycle on production of succinic acid in Saccharomyces cerevisiae . DB00139 is the main taste component produced by yeasts during sake ( Japanese rice wine ) fermentation . The pathway leading to accumulation of succinate was examined in liquid culture in the presence of a high concentration ( 15 % ) of glucose under aerobic and anaerobic conditions using a series of Saccharomyces cerevisiae strains in which various genes that encode the expression of enzymes required in TCA cycle were disrupted . When cultured in YPD medium containing 15 % glucose under aerobic conditions , the KGD1 ( alpha-ketoglutarate dehydrogenase ) gene disrupted mutant produced a lower level of succinate than the wild-type strain , while the P21912 ( succinate dehydrogenase ) gene-disrupted mutant produced an increased level of succinate . On the other hand , the FUM1 ( fumarase ) gene disrupted mutant produced significantly higher levels of fumarate but did not form malate at all . These results indicate that succinate , fumarate and malate are mainly synthesized through the TCA cycle ( oxidative direction ) even in the presence of glucose at a concentration as high as 15 % . When the growth condition was shifted from aerobic to anaerobic , the increased level of succinate in P21912 disruptants was no longer observed , whereas the decreased level of succinate in the KGD1 diruptant was still observed . A double mutant of the two fumarate reductase isozyme genes ( OSM1 and FRDS ) showed a succinate productivity of 50 % as compared to the parent when cells were incubated in glucose-buffered solution . These results indicate that succinate could be synthesized through two pathways , namely , alpha-ketoglutarate oxidation via the TCA cycle and fumarate reduction under anaerobic conditions . Effect of abiraterone acetate plus prednisone on the QT interval in patients with metastatic castration-resistant prostate cancer . PURPOSE : DB05812 is the active metabolite of the pro-drug abiraterone acetate ( AA ) and a selective inhibitor of P05093 , a key enzyme in testosterone synthesis , and improves overall survival in postdocetaxel metastatic castration-resistant prostate cancer ( mCRPC ) . This open-label , single-arm phase 1b study was conducted to assess the effect of AA and abiraterone on the QT interval . METHODS : The study was conducted in 33 patients with mCRPC . Patients received AA 1,000 mg orally once daily + prednisone 5 mg orally twice daily . Electrocardiograms ( ECGs ) were collected in triplicate using 12-lead Holter monitoring . Baseline ECGs were obtained on Cycle 1 Day-1 . Serial ECG recordings and time-matched pharmacokinetic ( PK ) blood samples were collected over 24 h on Cycle 1 Day 1 and Cycle 2 Day 1 . Serial PK blood samples were also collected over 24 h on Cycle 1 Day 8 . RESULTS : After AA administration , the upper bound of the 2-sided 90 % confidence interval ( CI ) for the mean baseline-adjusted QTcF change was < 10 ms ; no patients discontinued due to QTc prolongation or adverse events . No apparent relationship between change in QTcF and abiraterone plasma concentrations was observed [ estimated slope ( 90 % CI ) : 0.0031 ( -0.0040 , 0.0102 ) ] . CONCLUSIONS : There is no significant effect of AA plus prednisone on the QT/QTc interval in patients with mCRPC . 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 ) . 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 . Electron transfer is activated by calmodulin in the flavin domain of human neuronal nitric oxide synthase . The objective of this study was to clarify the mechanism of electron transfer in the human neuronal nitric oxide synthase ( P29475 ) flavin domain using the recombinant human P29475 flavin domains , the DB03147 /NADPH domain ( contains DB03147 - and NADPH-binding sites ) , and the DB03147 / Q68DA7 domain ( the flavin domain including a calmodulin-binding site ) . The reduction by NADPH of the two domains was studied by rapid-mixing , stopped-flow spectroscopy . For the DB03147 /NADPH domain , the results indicate that DB03147 is reduced by NADPH to generate the two-electron-reduced form ( FADH(2) ) and the reoxidation of the reduced DB03147 proceeds via a neutral ( blue ) semiquinone with molecular oxygen or ferricyanide , indicating that the reduced DB03147 is oxidized in two successive one-electron steps . The neutral ( blue ) semiquinone form , as an intermediate in the air-oxidation , was unstable in the presence of O(2) . The purified DB03147 /NADPH domain prepared under our experimental conditions was activated by NADP(+) but not NAD(+) . These results indicate that this domain exists in two states ; an active state and a resting state , and the enzyme in the resting state can be activated by NADP(+) . For the DB03147 / Q68DA7 domain , the reduction of the DB03147 - Q68DA7 pair of the oxidized enzyme with NADPH proceeded by both one-electron equivalent and two-electron equivalent mechanisms . The formation of semiquinones from the DB03147 - Q68DA7 pair was greatly increased in the presence of Ca(2+)/ P62158 . The air-stable semiquinone form , DB03147 -FMNH(.) , was further rapidly reduced by NADPH with an increase at 520 nm , which is a characteristic peak of the DB03147 semiquinone . Results presented here indicate that intramolecular one-electron transfer from DB03147 to Q68DA7 is activated by the binding of Ca(2+)/ P62158 . 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 . Evidence of increased microvessel density and activation of the hypoxia pathway in tumours from the hereditary leiomyomatosis and renal cell cancer syndrome . The Mendelian tumour syndromes hereditary leiomyomatosis and renal cell cancer ( HLRCC ) and hereditary paragangliomatosis with phaeochromocytomas ( HPGL ) result from mutations in nuclear genes ( FH and P21912 /C/D , respectively ) that encode Krebs cycle enzymes . HPGL tumours are highly vascular and there is evidence that inactivation of SDH leads to activation of the hypoxia/angiogenesis pathway . In contrast , uterine leiomyomas are not generally regarded as particularly vascular lesions . In order to test the possibility that activation of the hypoxia/angiogenesis pathway contributes to tumourigenesis in HLRCC , increased vascularity and hypoxia pathway activation were searched for in HLRCC tumours . Microvessel density was markedly higher in uterine leiomyomas from HLRCC than in the surrounding myometrium ; it was notable that sporadic uterine leiomyomas were actually less vascular than normal myometrium . In HLRCC tumours , there was increased expression of transcripts from the hypoxia-responsive genes vascular endothelial growth factor ( P15692 ) and Q12983 ; sporadic uterine leiomyomas did not show these changes . All uterine leiomyomas showed decreased expression of thrombospondin 1 . Although sporadic and HLRCC uterine leiomyomas appear to have identical morphology , their pathways of tumourigenesis may be fundamentally different . As is the case in HPGL , it is probable that failure of the Krebs cycle in HLRCC tumours causes inappropriate signalling that the cell is in a hypoxic state , leading to angiogenesis and perhaps directly to clonal expansion and tumour growth through some uncharacterized , cell-autonomous effect . Localization of somatostatin receptor subtype mRNA in the rat gastrointestinal tract and regulation of P30872 gene expression . Somatostatin and its receptors are widely distributed in the central nervous system and peripheral tissues including those of the gastrointestinal tract ( GI tract ) . The expression patterns of the five known SSTR genes have been analysed in detail by reverse transcription polymerase chain reaction amplifications and in situ hybridizations using tissues dissected from different parts of rat stomach and gut . While P30872 mRNA is present at relatively high amounts throughout the gastrointestinal tract , the levels of P30874 , 3 and 4 mRNAs vary in different regions and P35346 mRNA has not been detected . In situ hybridizations revealed the presence of P32745 mRNA in enterocytes and in neurons of the myenteric and submucous plexus . These findings are consistent with a role of P32745 in the observed somatostatin-mediated inhibition of acetylcholine release from myenteric neurons and of secretomotor neuron activity in the submucous plexus . Sequence analyses of the P30872 gene promoter revealed the absence of the canonical TATA and CAAT motifs and the presence of a variety of potential binding sites for transcriptional regulators . Among these are binding sites for P16383 , P05549 , AP-4 , response elements for somatostatin ( Q8TE85 -RE ) , epidermal growth factor ( P01133 -RE ) and cytocines ( GAS and NFIL ) as well as for tissue-specific factors such as Pit-1 ( pituitary ) and P52945 ( pancreatic cells ) . Mobility shift assays have confirmed that nuclear proteins of pancreatic RIN1046-38 and pituitary GH3 tumour cells bind to oligonucleotides containing the overlapping Pit-1 and P52945 binding sites . Thus , the Pit-1/ P52945 sites may be critical for the activation of the P30872 gene in these cell-types . DB01088 has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 + T cells was measured by ELISA . The expression of the transcription factor FoxP3 after co-culture of DCs with P01730 + CD45RA+ T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 , P05231 , P10145 and IL-12p70 in mDCs , while it enhanced P22301 production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA+ T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP3 expression and P22301 production . Additionally , iloprost inhibited the MIP-3beta-induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans . Microglia cyclooxygenase-2 activity in experimental gliomas : possible role in cerebral edema formation . PURPOSE : Cerebral edema is responsible for significant morbidity and mortality in patients harboring malignant gliomas . To examine the role of inflammatory cells in brain edema formation , we studied the expression cyclooxygenase ( P36551 ) -2 , a key enzyme in arachidonic acid metabolism , by microglia in the P13671 rodent glioma model . EXPERIMENTAL DESIGN : The expression of P35354 in primary microglia cultures obtained from intracranial rat P13671 gliomas was examined using reverse transcription-PCR , Western analysis , and prostaglandin E(2) ( PGE(2) ) enzyme immunoassay . Blood-tumor barrier permeability was studied in the same tumor model using magnetic resonance imaging . RESULTS : In contrast to P13671 glioma cells , microglia isolated from intracranial P13671 tumors produced high levels of PGE(2) through a P35354 -dependent pathway . To test whether the observed microglia P35354 activity played a role in brain edema formation in gliomas , tumor-bearing rats were treated with rofecoxib , a selective P35354 inhibitor . DB00533 was as effective as dexamethasone in decreasing the diffusion of contrast material into the brain parenchyma ( P = 0.01 , rofecoxib versus control animals ) , suggesting a reduction in blood-tumor barrier permeability . CONCLUSIONS : These findings suggest that glioma-infiltrating microglia are a major source of PGE(2) production through the P35354 pathway and support the use of P35354 inhibitors as possible alternatives to glucocorticoids in the treatment of peritumoral edema in patients with malignant brain tumors . 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 . [ 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 . P00747 activator dependent pathways in the dissemination of human tumor cells in the chick embryo . We have previously shown that inhibition of uPA activity of a human tumor-HEp3-results in a drastic reduction of its metastasis in the chick embryo . Using 125IUdR-labeled tumor cells , we have now studied the role of uPA in individual steps of tumor metastasis . We found that , 48 hr after inoculation of tumor cells on the P62158 , the organs of the embryos , inoculated with cells in which uPA was inhibited , contained 4-fold less cells than the controls . Neither the initial advance of the tumor mass into the P62158 nor the process of extravasation was affected by the inhibition of tumor uPA . However , the infiltration of the P62158 mesenchyme by individual tumor cells was blocked when tumor uPA activity or production was inhibited . In addition , indirect evidence implicated uPA as an essential factor in the tumor cell intravasation . Battle of the kinases : integration of adrenal responses to DB02527 , DG and Ca2+ at the level of steroidogenic cytochromes P450 and 3betaHSD expression in H295R cells . While DB01285 receptors ( activating the protein kinase A pathway ) are expressed throughout the human/bovine/ovine zona glomerulosa ( zg ) and zona fasciculata ( zf ) , there are clear zonal differences in AII Type-1 receptor levels ( activating protein kinase C/Ca2+ ) , as well as resting membrane potential . Thus zg is most responsive to AII and K+ ( Ca2+ signalling ) , while zf is less responsive to AII with no K+ response . Zonal function in turn requires differential expression of P05093 /3betaHSD and P19099 / P15538 . We have used the H295R cell to study how differential activation of kinase A , kinase C and Ca2+/calmodulin ( P62158 ) kinases may alter the relative expression of the steroidogenic P450s and 3betaHSDII . While P05108 , P05093 , 3betaHSDII , P08686 , and P15538 are all induced by increases in DB02527 , studies with TPA alone or in combination with forskolin reveal subsets of steroidogenic enzymes regulated either positively ( P08686 , 3betaHSDII ) or negatively ( P05093 , P05108 ) by protein kinase C . Thus adrenal 3betaHSDII and P08686 expression is high in zg and zf , but P05093 is not expressed in the zg where AII activation of kinase C is highest . In turn both K+ and AII-induced elevation of Ca2+ strongly induces P19099 but not P15538 , consistent with preferential expression of P19099 in the zg . We conclude that differential signaling through kinase C and P62158 kinases in addition to kinase A underlies zonal differences in both the early and late pathways involved in steroid hormone production within the adrenocortical zones . 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 . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . 5-hydroxytryptamine evokes endothelial nitric oxide synthase activation in bovine aortic endothelial cell cultures . Activation of endothelial nitric oxide synthase ( P29474 ) results in the production of nitric oxide ( NO ) that mediates the vasorelaxing properties of endothelial cells . The goal of this project was to address the possibility that 5-hydroxytryptamine ( 5-HT ) stimulates P29474 activity in bovine aortic endothelial cell ( BAEC ) cultures . Here , we tested the hypothesis that 5-HT receptors mediate P29474 activation by measuring agonist-stimulated [3H]L-citrulline ( [3H]L- DB00155 ) formation in BAEC cultures . We found that 5-HT stimulated the conversion of [3H]L-arginine ( [3H] DB00125 ) to [3H]L- DB00155 , indicating P29474 activation . The high affinity P28222 receptor agonist , 5-nonyloxytryptamine ( 5-NOT ) -stimulated [3H]L- DB00155 turnover responses were concentration- ( 0.01 nM to 100 microM ) and time-dependent . Maximal responses were observed within 10 min following agonist exposures . These responses were effectively blocked by the P28222 receptor antagonist , isamoltane , the P28222 /5-HT2 receptor antagonist , methiothepin , and the P29474 selective antagonists ( 0.01-10 microM ) : L-Nomega -monomethyl-L-arginine ( L-NMMA ) and L-N omega-iminoethyl-L-ornithine ( L-NIO ) . Pretreatment of BAEC cultures with pertussis toxin ( PTX ; 1-100 ng/ml ) for 16 hr resulted in significant inhibition of the agonist-stimulated P29474 activity , indicating the involvement of Gi proteins . These findings lend evidence of a P28222 receptor/ P29474 pathway , accounting in part for the activation of P29474 by 5-HT . Further investigation is needed to determine the role of other vascular 5-HT receptors in the stimulation of P29474 activity . P62158 dependence of somatostatin release stimulated by growth hormone-releasing factor . Experiments were performed in vitro to examine the possible role of calcium and calmodulin in P01286 -induced somatostatin ( SRIF ) release from the median eminence . Adult male rats were used as tissue donors . The median eminences were first prestimulated in 0.4 ml Krebs Ringer bicarbonate glucose buffer ( pH 7.4 ) containing bacitracin at 37C in an atmosphere of 95 % O2 , 5 % CO2 with constant shaking for 30 min . When calcium was omitted , this medium was used during the prestimulation and stimulation periods . After prestimulation , the medium was discarded and replaced by medium containing the different substances to be tested ( P01286 , EGTA , calcium channel blockers , and calmodulin inhibitors ) . The stimulation of SRIF release induced by 10(-10) M P01286 was not inhibited by omission of extracellular calcium or when the remaining CA+2 was chelated with 10(-4) M EGTA . The calcium channel blockers , nifendipine and verapamil ( 10(-6) M ) , failed to alter the increase of SRIF release induced by rGRF . Three calmodulin inhibitors were employed to examine the possible influence of calmodulin on P01286 -induced SRIF release . Trifluoperazine ( 10(-6) M ) , triflupromazine ( 10(-6) M ) and penfluridol ( 10(-7) M ) had an inhibitory effect on the stimulation of SRIF release induced by P01286 and failed to alter resting release . Thus , P01286 can evoke SRIF release independently of extraterminal Ca+2 concentration and Ca+2 influx into the nerve terminals , but the releasing process involves translocation of Ca+2 from intracellular stores . The inhibitory effect of the calmodulin inhibitors on P01286 -induced SRIF release , suggests that the translocated Ca+2 must bind to calmodulin in order to release SRIF .

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interacts_with DB00977?

multiple_choice

DB00197 sensitizes tumor cells to P50591 -induced apoptosis via down-regulation of FLIP and Survivin . Induction of apoptosis by the death ligand P50591 might be a promising therapeutic approach in cancer therapy . However , since not all tumor cells are sensitive to P50591 , there is a need for the development of strategies to overcome P50591 -resistance . The results of the present study show that the anti-diabetic drug troglitazone sensitizes human glioma and neuroblastoma cells to P50591 -induced apoptosis . This process is accompanied by a substantial increase of active caspase 8 and active caspase 3 , but it is independent of troglitazone 's effects on the nuclear receptor P37231 . DB00197 induces a pronounced reduction in protein expression levels of the anti-apoptotic Q14790 -inhibitory protein ( FLIP ) without affecting FLIP mRNA levels . Further , protein and mRNA expression levels of the anti-apoptotic protein Survivin significantly decrease upon treatment with troglitazone . Moreover , sensitization to P50591 is partly accompanied by an up-regulation of the P50591 receptor , O14763 . A combined treatment with troglitazone and P50591 might be a promising experimental therapy because troglitazone sensitizes tumor cells to P50591 -induced apoptosis via various mechanisms , thereby minimizing the risk of acquired tumor cell resistance . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . 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 . 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 . 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 . 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 . Effect of exercise on atrial natriuretic peptide ( P01160 ) levels in patients with P48444 . A possible role of mitochondria in the apoptotic-like programmed nuclear death of Tetrahymena thermophila . The ciliated protozoan Tetrahymena has a unique apoptosis-like process , which is called programmed nuclear death ( P01160 ) . During conjugation , the new germinal micro- and somatic macro-nuclei differentiate from a zygotic fertilized nucleus , whereas the old parental macronucleus degenerates , ensuring that only the new macronucleus is responsible for expression of the progeny genotype . As is the case with apoptosis , this process encompasses chromatin cleavage into high-molecular mass DNA , oligonucleosomal DNA laddering , and complete degradation of the nuclear DNA , with the ultimate outcome of nuclear resorption . Q14790 - and caspase-9-like activities are involved in the final resorption process of P01160 . In this report , we show evidence for mitochondrial association with P01160 . Mitochondria and the degenerating macronucleus were colocalized in autophagosome using two dyes for the detection of mitochondria . In addition , an endonuclease with similarities to mammalian endonuclease G was detected in the isolated mitochondria . When the macronuclei were incubated with isolated mitochondria in a cell-free system , DNA fragments of 150-400 bp were generated , but no DNA ladder appeared . Taking account of the present observations and the timing of autophagosome formation , we conclude that mitochondria might be involved in Tetrahymena P01160 , probably with the process of oligonucleosomal laddering . Establishment by adriamycin exposure of multidrug-resistant rat ascites hepatoma AH130 cells showing low P15559 activity and high cross resistance to mitomycins . A resistant subline ( AH130/5A ) selected from rat hepatoma AH130 cells after exposure to adriamycin ( P35318 ) showed remarkable resistance to multiple antitumor drugs , including mitomycin C ( DB00305 ) and porfiromycin ( PFM ) . PFM , vinblastine ( DB00570 ) , and P35318 accumulated in AH130/5A far less than in the parent AH130 ( AH130/P ) cells . AH130/5A cells showed overexpression of P-glycoprotein ( A6NDG6 ) , an increase in glutathione S-transferase activity , and a decrease in P15559 and glutathione peroxidase activity . The resistance to DB00305 and DB00570 of AH130/5A cells was partly reversed by H-87 , an inhibitor of A6NDG6 . Buthionine sulfoximine , an inhibitor of glutathione synthase , did not affect the action of DB00305 . tert-Butylhydroquinone induced P15559 activity , increased PFM uptake , and enhanced the growth-inhibitory action of DB00305 in AH130/5A cells . DB00266 , an inhibitor of P15559 , decreased PFM uptake and reduced the growth-inhibitory action of DB00305 in AH130/P cells . These results indicated that the adriamycin treatment of hepatoma cells caused multifactorial multidrug resistance involving a decrease in P15559 activity . Differential effects of the vascular endothelial growth factor receptor inhibitor PTK787/ZK222584 on tumor angiogenesis and tumor lymphangiogenesis . Halting tumor growth by interfering with tumor-induced angiogenesis is an attractive therapeutic approach . Such treatments include humanized antibodies blocking the activity of vascular endothelial growth factor ( P15692 ) -A ( bevacizumab ) , soluble P15692 receptor ( VEGFR ) constructs ( P15692 - Q8NHU6 ) , or small-molecule inhibitors of VEGFR signaling , including PTK787/ZK222584 ( DB04879 ) , sorafenib , and sunitinib . DB04879 has been shown previously to specifically block P15692 -induced phosphorylation of P17948 , -2 and -3 and thereby to inhibit endothelial cell proliferation , differentiation , and tumor angiogenesis . We have investigated the effect of DB04879 on tumor angiogenesis and tumor lymphangiogenesis using the Rip1Tag2 transgenic mouse model of pancreatic beta cell carcinogenesis . In Rip1Tag2 mice , tumor angiogenesis is predominantly mediated by P15692 , and as expected , DB04879 efficiently impaired tumor blood vessel angiogenesis and tumor growth . Double-transgenic Rip1Tag2;Rip1VEGF-C and Rip1Tag2;Rip1VEGF-D mice not only exhibit P15692 -dependent blood vessel angiogenesis but also tumor lymphangiogenesis induced by the transgenic expression of P49767 or -D . In these mouse models , DB04879 also repressed tumor blood vessel angiogenesis and tumor growth yet failed to affect tumor lymphangiogenesis and lymphogenic metastasis . Adenoviral delivery of soluble P35916 also did not prevent tumor lymphangiogenesis in these mice . In contrast , spontaneous tumor lymphangiogenesis , as observed by the stochastic expression of P49767 and -D in tumors of neural cell adhesion molecule-deficient Rip1Tag2 mice , was repressed by DB04879 and soluble P35916 . The results indicate that the time of onset and the levels of P49767 /D expression may be critical variables in efficiently repressing tumor lymphangiogenesis and that pathways other than VEGFR signaling may be involved in tumor lymphangiogenesis . Adrenomedullin modulates the neurohumoral response to acute volume loading in normal conscious sheep . The physiological role of adrenomedullin ( P35318 ) in volume and pressure homeostasis remains unclear . Accordingly , we assessed possible modulatory actions of P35318 infusions on the neurohumoral response to acute volume loading with dextran in normal conscious sheep . DB09255 ( 15 ml/kg ) , given with concurrent P35318 ( 5.5 pmol/kg per min -- raising plasma P35318 from below detection to approximately 10 pmol/l ) or vehicle control infusions , induced matched significant ( P < 0.001 by Q9UNW9 ) falls in hematocrit ( 27-30 % ) during both P35318 and control and similar increases in right atrial pressure ( approximately 10 mmHg ) . Compared with control , both systemic ( P=0.033 ) and pulmonary ( P=0.005 ) arterial pressure and peripheral resistance ( P=0.004 ) were reduced during P35318 but were raised post-infusion . The dextran-induced increase in cardiac output was augmented by P35318 ( P=0.048 ) . DB09255 -induced increases in plasma atrial natriuretic peptide ( P01160 ; P=0.008 ) , brain natriuretic peptide ( DB04899 ; P=NS ) and cyclic guanosine monophosphate ( cGMP ; P=0.003 ) were augmented post- P35318 infusions . The dextran-induced fall in plasma renin activity ( P06703 ) was attenuated by P35318 ( P=0.039 ) whereas plasma aldosterone levels were unaltered . P35318 augmented the increase in urinary volume during the second 2-h clearance period post-dextran . Our data indicate that P35318 modifies the hemodynamic and hormonal response to an acute volume challenge , enhances natriuretic peptide secretion and reduces systemic vascular resistance . These results provide further evidence that P35318 plays a physiological role in volume and pressure homeostasis . Epigenetic inactivation of the chromosomal stability control genes P38398 , P51587 , and P13010 in non-small cell lung cancer . PURPOSE : Lung cancer cells frequently exhibit marked chromosome instability . We postulated that alterations of the double-strand break repair genes ( P38398 , P51587 , and P13010 ) might be involved in lung cancer . PATIENTS AND METHODS : We examined the loss of protein and mRNA expression and the 5'CpG hypermethylation and allelic imbalance of the P38398 , P51587 , and P13010 genes in 98 non-small cell lung cancer ( NSCLC ) samples . Anchorage-dependent growth after reexpression of these genes was examined in a lung cancer cell line that originally lacked P38398 and P51587 expression . RESULTS : The data indicated that low protein expression of P38398 and P51587 was frequent in lung adenocarcinomas ( 42-44 % ) , whereas low P13010 protein expression was more prevalent among squamous cell carcinoma ( 32 % ) . In addition , low P38398 expression was significantly associated with low RB expression , especially in lung adenocarcinoma . Concurrent alterations in P13010 and p53 were the most frequent profiles in smoking patients . Importantly , low mRNA and protein expressions of P38398 , P51587 , and P13010 were significantly associated with their promoter hypermethylation . 5-Aza-2'-deoxycytidine treatment of NSCLC cells showed demethylation and reexpression of the P38398 and P51587 genes and reduced anchorage-independent growth . CONCLUSIONS : Our retrospective study provides compelling evidence that low mRNA and protein expression in the P38398 / P51587 and P13010 genes occur in lung adenocarcinoma and squamous cell carcinoma , respectively , and that promoter hypermethylation is the predominant mechanism in deregulation of these genes . Alteration of the double-strand break repair pathway , perhaps by interacting with p53 and RB deregulation , is important in the pathogenesis of a subset of NSCLC . Ku affects the ataxia and Rad 3-related/ O14757 -dependent S phase checkpoint response after camptothecin treatment . DB04690 ( CPT ) that targets P11387 is one of the most promising broad-spectrum anticancer drugs in development today . The cytotoxicity of CPT is S phase ( S ) -specific because the collision of advancing replication forks with CPT-topoisomerase I-DNA complexes results in DNA damage . After DNA damage , proliferating cells could actively slow down the DNA replication through an S checkpoint to provide time for repair . We report now that there is an activated S checkpoint response in CPT-treated mammalian cells . This response is regulated by Ataxia and Rad3-related ( ATR ) / O14757 pathway . Compared with their wild-type counterparts , CPT-treated P13010 -/- cells showed stronger inhibition of DNA replication . This stronger inhibition had no relationship with DNA-dependent protein kinase ( DNA-PK ) activity but correlated with the higher activities of ATR and the higher activities of O14757 in such cells . Not only caffeine , the nonspecific inhibitor of ATR , or P55089 -01 , the nonspecific inhibitor of O14757 , but also the specific O14757 antisense oligonucleotide abolished the stronger inhibition of DNA replication in CPT-treated P13010 -/- cells . These results in aggregate indicated that the stronger S checkpoint in CPT-treated P13010 -/- cells is regulated through the highly activated ATR/ O14757 pathway . Differential effects of protein kinase C , Ras , and P04049 kinase on the induction of the cardiac B-type natriuretic peptide gene through a critical promoter-proximal M-CAT element . The cardiac genes for the A- and B-type natriuretic peptides ( P01160 and DB04899 ) are coordinately induced by growth promoters , such as alpha1-adrenergic receptor agonists ( e.g. phenylephrine ( PE ) ) . Although inducible elements in the P01160 gene have been identified , responsible elements in the DB04899 gene are unknown . In this study , reporter constructs transfected into neonatal rat ventricular myocytes showed that in the context of 2.5 kilobase pairs of native DB04899 5'-flanking sequences , a 2-base pair mutation in a promoter-proximal M-CAT site ( CATTCT ) disrupted basal and PE-inducible transcription by more than 98 % . Expression of constitutively active forms of Ras , P04049 kinase , and protein kinase C , all of which are activated by PE in cardiac myocytes , strongly stimulated DB04899 reporter expression . Isolated M-CAT elements conferred PE , protein kinase C , and Ras inducibility to a minimal DB04899 promoter , however , they did not confer P04049 inducibility . These results show that M-CAT elements can serve as targets for Ras-dependent , P04049 -independent pathways , implying the involvement of c-Jun N-terminal kinase and/or p38 mitogen-activated protein kinases , but not extracellular signal-regulated protein kinase/mitogen-activated protein kinase . Moreover , the essential M-CAT element distinguishes the DB04899 gene from the P01160 gene , which utilizes serum response elements and an Sp1-like sequence . Molecular targets and regulators of cardiac hypertrophy . Cardiac hypertrophy is one of the main ways in which cardiomyocytes respond to mechanical and neurohormonal stimuli . It enables myocytes to increase their work output , which improves cardiac pump function . Although cardiac hypertrophy may initially represent an adaptive response of the myocardium , ultimately , it often progresses to ventricular dilatation and heart failure which is one of the leading causes of mortality in the western world . A number of signaling modulators that influence gene expression , apoptosis , cytokine release and growth factor signaling , etc. are known to regulate heart . By using genetic and cellular models of cardiac hypertrophy it has been proved that pathological hypertrophy can be prevented or reversed . This finding has promoted an enormous drive to identify novel and specific regulators of hypertrophy . In this review , we have discussed the various molecular signal transduction pathways and the regulators of hypertrophic response which includes calcineurin , cGMP , NFAT , natriuretic peptides , histone deacetylase , P05231 cytokine family , Gq/ P49842 signaling , PI3K , MAPK pathways , Na/H exchanger , DB01367 , polypeptide growth factors , P01160 , NO , P01375 , Q07869 and JAK/ P35610 pathway , microRNA , Cardiac angiogenesis and gene mutations in adult heart . Augmented knowledge of these signaling pathways and their interactions may potentially be translated into pharmacological therapies for the treatment of various cardiac diseases that are adversely affected by hypertrophy . The purpose of this review is to provide the current knowledge about the molecular pathogenesis of cardiac hypertrophy , with special emphasis on novel researches and investigations . Culture effects of epidermal growth factor ( P01133 ) and basic fibroblast growth factor ( P09038 ) on cryopreserved human adipose-derived stromal/stem cell proliferation and adipogenesis . Previous studies have demonstrated that P01133 and P09038 maintain the stem cell properties of proliferating human adipose-derived stromal/stem cells ( hASCs ) in vitro . While the expansion and cryogenic preservation of isolated hASCs are routine , these manipulations can impact their proliferative and differentiation potential . This study examined cryogenically preserved hASCs ( n = 4 donors ) , with respect to these functions , after culture with basic fibroblast growth factor ( P09038 ) and epidermal growth factor ( P01133 ) at varying concentrations ( 0-10 ng/ml ) . Relative to the control , cells supplemented with P01133 and P09038 significantly increased proliferation by up to three-fold over 7-8 days . Furthermore , cryopreserved hASCs expanded in the presence of P01133 and P09038 displayed increased oil red O staining following adipogenic induction . This was accompanied by significantly increased levels of several adipogenesis-related mRNAs : aP2 , C/EBPalpha , lipoprotein lipase ( P06858 ) , PPARgamma and PPARgamma co-activator-1 ( PGC1 ) . Adipocytes derived from P01133 - and P09038 -cultured hASCs exhibited more robust functionality based on insulin-stimulated glucose uptake and atrial natriuretic peptide ( P01160 ) -stimulated lipolysis . These findings indicate that P09038 and P01133 can be used as culture supplements to optimize the proliferative capacity of cryopreserved human ASCs and their adipogenic differentiation potential . 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 . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . 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 .

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Proteomic analysis of human fetal atria and ventricle . In this study we carried out a mass spectrometry-based proteome analysis of human fetal atria and ventricles . Heart protein lysates were analyzed on the Q-Exactive mass spectrometer in biological triplicates . Protein identification using MaxQuant yielded a total of 2754 atrial protein groups ( 91 % ) and 2825 ventricular protein groups ( 83 % ) in at least 2 of the 3 runs with ≥ 2 unique peptides . Statistical analyses using fold-enrichment ( > 2 ) and p-values ( ≤ 0.05 ) selected chamber-enriched atrial ( 134 ) and ventricular ( 81 ) protein groups . Several previously characterized cardiac chamber-enriched proteins were identified in this study including atrial isoform of myosin light chain 2 ( Q01449 ) , atrial natriuretic peptide ( P01160 ) , connexin 40 ( P36382 ) , and peptidylglycine alpha-amidating monooxygenase ( PAM ) for atria , and ventricular isoforms of myosin light chains ( P10916 and P08590 ) , myosin heavy chain 7 ( P12883 ) , and connexin 43 ( P17302 ) for ventricle . Our data was compared to in-house generated and publicly available human microarrays , several human cardiac proteomes , and phenotype ontology databases . Possible involvement of Q07869 gamma in the regulation of basal channel opening of Q99572 receptor in cultured mouse astrocytes . AIMS : Recently , we demonstrated that cultured mouse astrocytes exhibited basal channel opening of Q99572 receptor ( P2X7R ) in the absence of any exogenous ligand , but the regulatory mechanism involved was not elucidated . Since our preliminary experiments suggested possible involvement of peroxisome proliferator-activated receptor ( Q07869 ) gamma in the regulation , we examined whether Q07869 gamma regulated P2X7R basal channel opening in mouse astrocytes . MAIN METHODS : P2X7R channel opening was assessed as to the uptake of a marker dye , YO-PRO-1 ( YP ) , in the presence or absence of agonists and antagonists for Q07869 gamma under a fluorescence microscope . Expression of Q07869 gamma was evaluated by Western blotting and immunocytochemistry . KEY FINDINGS : NSAIDs such as flufenamic acid ( FFA ) and indomethacin , which are a cyclooxygenase inhibitor and a Q07869 gamma agonist , showed enhancing and inhibiting effects on YP uptake at low and high concentrations , respectively , and the enhanced uptake was abolished by periodate-oxidized DB00171 ( oxATP ) , a selective P2X7R antagonist . The Q07869 gamma agonists 15-deoxy-Delta(12,14)-prostaglandin J(2) and ciglitazone decreased the basal and FFA-enhanced YP uptake , while the antagonist GW9662 increased YP uptake , this effect being blocked by the agonists and also by oxATP . Q07869 gamma was distributed in the nucleus and cytosolic/membrane fraction of cultured mouse astrocytes . SIGNIFICANCE : These findings indicate that basal channel opening of P2X7R in mouse astrocytes is at least in part regulated by Q07869 gamma . Plasma n-3 fatty acid response to an n-3 fatty acid supplement is modulated by apoE epsilon4 but not by the common Q07869 L162V polymorphism in men . The risk of Alzheimer 's disease is increased for carriers of apoE4 ( E4 ) or the Q07869 L162V polymorphism ( L162V ) , but it is decreased in fish and seafood consumers . The link between high fish intake and reduced risk of cognitive decline in the elderly appears not to hold in carriers of E4 , possibly because better cognition is linked to EPA+ DB01708 in the blood , but only in non-carriers of E4 . As yet , no such studies exist in carriers of L162V . Our objective was to determine whether the plasma fatty acid response to a dietary supplement of EPA+ DB01708 was altered in carriers of L162V and/or E4 . This was an add-on project ; in the original study , men were selected based on whether or not they were carriers of L162V ( n 14 per group ) . E4 status was determined afterwards . All subjects received an EPA+ DB01708 supplement for 6 weeks . L162V polymorphism did not interact with the supplement in a way to alter EPA and DB01708 incorporation into plasma lipids . However , when the groups were separated based on the presence of E4 , baseline EPA and DB01708 in plasma TAG were 67 and 60 % higher , respectively , in E4 carriers . After the supplementation , there were significant gene x diet interactions in which only non-carriers had increased EPA and DB01708 in plasma P80303 and TAG , respectively . 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 . Q99572 receptor-dependent intestinal afferent hypersensitivity in a mouse model of postinfectious irritable bowel syndrome . The DB00171 -gated P2X(7) receptor ( P2X(7)R ) was shown to be an important mediator of inflammation and inflammatory pain through its regulation of IL-1β processing and release . Trichinella spiralis-infected mice develop a postinflammatory visceral hypersensitivity that is reminiscent of the clinical features associated with postinfectious irritable bowel syndrome . In this study , we used P2X(7)R knockout mice ( P2X(7)R(-/-) ) to investigate the role of P2X(7)R activation in the in vivo production of IL-1β and the development of postinflammatory visceral hypersensitivity in the T. spiralis-infected mouse . During acute nematode infection , IL-1β-containing cells and P2X(7)R expression were increased in the jejunum of wild-type ( WT ) mice . Peritoneal and serum IL-1β levels were also increased , which was indicative of elevated IL-1β release . However , in the P2X(7)R(-/-) animals , we found that infection had no effect upon intracellular , plasma , or peritoneal IL-1β levels . Conversely , infection augmented peritoneal P01375 -α levels in both WT and P2X(7)R(-/-) animals . Infection was also associated with a P2X(7)R-dependent increase in extracellular peritoneal lactate dehydrogenase , and it triggered immunological changes in both strains . Jejunal afferent fiber mechanosensitivity was assessed in uninfected and postinfected WT and P2X(7)R(-/-) animals . Postinfected WT animals developed an augmented afferent fiber response to mechanical stimuli ; however , this did not develop in postinfected P2X(7)R(-/-) animals . Therefore , our results demonstrated that P2X(7)Rs play a pivotal role in intestinal inflammation and are a trigger for the development of visceral hypersensitivity . Involvement of the Q99572 purinergic receptor and c-Jun N-terminal and extracellular signal-regulated kinases in cyclooxygenase-2 and prostaglandin E2 induction by LL-37 . Periodontal disease is caused by microorganisms and host-derived inflammation involving increased cyclooxygenase-2 ( P35354 ) expression and prostaglandin E(2) ( PGE(2) ) production . We previously demonstrated that human β-defensin-3 induces P35354 and PGE(2) in human gingival fibroblasts ( HGFs ) . We , therefore , aimed to examine the inducible effects of LL-37 , the only cathelicidin expressed in humans , on P35354 expression and PGE(2) synthesis in HGFs and to elucidate the relevant signaling pathways . The P35354 expression was upregulated by LL-37 in dose- and time-dependent manners . Accordingly , the synthesis of PGE(2) in cell-free culture supernatants was raised by LL-37 ( p < 0.01 ) and blocked by NS-398 , a specific P35354 inhibitor ( p < 0.01 ) . P2X inhibitors and a neutralizing antibody against P2X(7) purinergic receptor significantly abrogated P35354 induction and PGE(2) production by LL-37 ( p < 0.01 ) . LL-37 upregulated P35354 expression and PGE(2) synthesis via activation of extracellular signal-regulated kinase ( P29323 ) and Q9BY77 c-Jun N-terminal kinase ( JNK ) , while interleukin-1β did so via nuclear factor-ĸB and all three mitogen-activated protein kinases . In summary , LL-37 can control arachidonic acid metabolism by induction of P35354 expression and PGE(2) synthesis via the P2X(7) receptor , P29323 , and Q9BY77 JNK . The pro-inflammatory effects of LL-37 may be essential for initiating oral mucosal inflammation in periodontal disease . Supramolecular signalling complexes in the nervous system . It is now apparent that multiprotein signalling complexes or " signalling machines " are responsible for orchestrating many complex signalling pathways in the cell . The synapse is a sub-cellular specialisation which transmits and converts patterns of electrical activity into cellular memory . This processing of electrical information is mediated by the protein components of the synapse . The organisation of synaptic proteins has been investigated over the last number of years using proteomic methods and with the application ofbioinformatics ; a landscape of modular protein complexes at the synapse is emerging . Many share a common organisation centred on a receptor/channel , a protein scaffold , ( in which the signalling molecules are localised ) and membrane to cytoskeleton interactions . The use of PDZ-domain based protein scaffolds is a particularly common feature in the construction of neuronal protein complexes and the differential presence of these proteins in complexes can have functional consequences . Here we overview current proteomic methodologies for the analysis of multiprotein complexes . In addition , we describe the characterisation of a number of multiprotein complexes associated with ion channels ( NMDAR , Q99572 and Kir2 ) and GPCRs ( 5- Q13049 / P28335 , D2 and P41594 ) and discuss common their common components and organisation . Autocrine regulation of γ-irradiation-induced DNA damage response via extracellular nucleotides-mediated activation of Q15077 and Q9H244 receptors . A key component of the response to DNA damage caused by ionizing radiation is DNA repair . Release of extracellular nucleotides , such as DB00171 , from cells plays a role in signaling via P2 receptors . We show here that release of DB00171 , followed by activation of P2Y receptors , is involved in the response to γ-irradiation-induced DNA damage . Formation of phosphorylated histone variant P16104 ( γ P16104 ) foci , which are induced in nuclei by DNA damage and contribute to accumulation of DNA-repair factors , was increased at 1-3h after γ-ray irradiation ( 2.0Gy ) of human lung cancer A549 cells . Focus formation was suppressed by pre-treatment with the ecto-nucleotidase apyrase . Pre-treatment with ecto-nucleotidase inhibitor ARL67156 or post-treatment with DB00171 or UTP facilitated induction of γ P16104 , indicating that extracellular nucleotides play a role in induction of γ P16104 foci . Next , we examined the effect of P2 receptor inhibitors on activation of ataxia telangiectasia mutated ( Q13315 ; a protein kinase ) and accumulation of Q12888 ( a DNA repair factor ) , both of which are important for DNA repair , at DNA damage sites . Q15077 receptor antagonist MRS2578 , Q9H244 receptor antagonist clopidogrel , and Q99572 receptor antagonists A438079 and oxATP significantly inhibited these processes . Release of DB00171 was detected within 2.5min after irradiation , but was blocked by A438079 . Activation of Q13315 and accumulation of Q12888 were decreased in Q15077 or Q9H244 receptor-knockdown cells . We conclude that autocrine/paracrine signaling through Q99572 -dependent DB00171 release and activation of Q15077 and Q9H244 receptors serves to amplify the cellular response to DNA damage caused by γ-irradiation . 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 . Osteoblastic protein tyrosine phosphatases inhibition and connexin 43 phosphorylation by alendronate . Bisphosphonates ( BPs ) , potent inhibitors of bone resorption which inhibit osteoclasts , have also been shown to act on osteocytes and osteoblasts preventing apoptosis via connexin ( Cx ) 43 hemichannels and activating the extracellular signal regulated kinases ERKs . We previously demonstrated the presence of a saturable , specific and high affinity binding site for alendronate ( ALN ) in osteoblastic cells which express P17302 . However , cells lacking P17302 also bound BPs . Herein we show that bound [ (3)H ] -alendronate is displaced by phosphatase substrates . Moreover , similar to Na3VO4 , ALN inhibited the activity of transmembrane and cytoplasmic PTPs , pointing out the catalytic domain of phosphatases as a putative BP target . In addition , anti-phospho-tyrosine immunoblot analysis revealed that ALN stimulates tyrosine phosphorylation of several proteins of whole cell lysates , among which the major targets of the BP could be immunochemically identified as P17302 . Additionally , the transmembrane receptor-like PTPs , RPTPµ and RPTPα , as well as the cytoplasmic P18031 , are highly expressed in ROS 17/2.8 cells . Furthermore , we evidenced that P17302 interacts with RPTPµ in ROS 17/2.8 and ALN decreases their association . These results support the hypothesis that BPs bind and inhibit PTPs associated to P17302 or not , which would lead to the activation of signaling pathways in osteoblasts . Connexin36 knockout mice display increased sensitivity to pentylenetetrazol-induced seizure-like behaviors . OBJECTIVE : Large-scale synchronous firing of neurons during seizures is modulated by electrotonic coupling between neurons via gap junctions . To explore roles for connexin36 ( Q9UKL4 ) gap junctions in seizures , we examined the seizure threshold of connexin36 knockout ( Cx36KO ) mice using a pentylenetetrazol ( PTZ ) model . METHODS : Mice ( 2-3months old ) with Q9UKL4 wildtype ( WT ) or Cx36KO genotype were treated with vehicle or 10-40mg/kg of the convulsant PTZ by intraperitoneal injection . Seizure and seizure-like behaviors were scored by examination of video collected for 20min . Quantitative real-time PCR ( QPCR ) was performed to measure potential compensatory neuronal connexin ( Q8NFK1 , P35212 , P17302 and P36383 ) , pannexin ( Q96RD7 and Q96RD6 ) and gamma-aminobutyric acid type A ( GABA(A) ) receptor α1 subunit gene expression . RESULTS : Cx36KO animals exhibited considerably more severe seizures ; 40mg/kg of PTZ caused severe generalized ( ≥grade III ) seizures in 78 % of KO , but just 5 % of WT mice . A lower dose of PTZ ( 20mg/kg ) induced grade II seizure-like behaviors in 40 % KO vs. 0 % of WT animals . There was no significant difference in either connexin , pannexin or GABA(A) α1 gene expression between WT and KO animals . CONCLUSION : Increased sensitivity of Cx36KO animals to PTZ-induced seizure suggests that Q9UKL4 gap junctional communication functions as a physiological anti-convulsant mechanism , and identifies the Q9UKL4 gap junction as a potential therapeutic target in epilepsy . Topoisomerase II-mediated DNA cleavage and mutagenesis activated by nitric oxide underlie the inflammation-associated tumorigenesis . AIMS : Both cancer-suppressing and cancer-promoting properties of reactive nitrogen and oxygen species ( RNOS ) have been suggested to play a role in tumor pathology , particularly those activities associated with chronic inflammation . Here , we address the impact of nitric oxide ( NO ) on the induction of DNA damage and genome instability with a specific focus on the involvement of topoisomerase II ( P11388 ) . We also investigate the contribution of NO to the formation of skin melanoma in mice . RESULTS : Similar to the P11388 -targeting drug , etoposide ( DB00773 ) , the NO-donor , S-nitrosoglutathione ( GSNO ) , induces skin melanomas formation in 7,12-dimethyl- benz[a]anthracene ( DMBA ) -initiated mice . To explore the mechanism(s) underlying this NO-induced tumorigenesis , we use a co-culture model system to demonstrate that inflamed macrophages with inducible NO synthase ( P35228 ) expression cause γ- P16104 activation , p53 phosphorylation , and chromosome DNA breaks in the target cells . Inhibitor experiments revealed that NO and P11388 isozymes are responsible for the above described cellular phenotypes . Notably , NO , unlike DB00773 , preferentially induces the formation of TOP2β cleavable complexes ( TOP2βcc ) in cells . Moreover , GSNO induced P11388 -dependent DNA sequence rearrangements and cytotoxicity . Furthermore , the incidences of GSNO- and DB00773 -induced skin melanomas were also observed to be lower in the skin-specific top2β-knockout mice . Our results suggest that P11388 isozymes contribute to NO-induced mutagenesis and subsequent cancer development during chronic inflammation . INNOVATION AND CONCLUSIONS : We provide the first experimental evidence for the functional role of P11388 in NO-caused DNA damage , mutagenesis , and carcinogenesis . Notably , these studies contribute to our molecular understanding of the cancer-promoting actions of RNOS during chronic inflammation . [ Human monocyte-derived multinucleated giant cells ] . Multinucleated giant cells ( MGC ) are characteristic cells in granulomatous disorders such as sarcoidosis and leprosy . There are two types of MGC ; foreign body-type and Langhans-type cells . The exact mechanisms of the formation and the functional significance of MGC are not determined , although their morphological features are well understood . MGC are also formed in vitro from peripheral blood mononuclear cells by stimulation with cytokines and lectins . Particularly P01579 is considered to play a pivotal role in monocyte fusion . P08700 , P05112 , P35225 , and GM- P04141 are other reported cytokines involved in MGC formation . In addition to such inflammatory mediators , a factor derived from the pathogens of granulomatous disorders may be necessary for MGC formation . Muramyl dipeptide , a peptidoglycan portion of bacterial cell walls , is one of the candidates and can preferentially induce Langhans-type cells in in vitro MGC formation system . Although the exact mechanisms of in vitro MGC formation remains unknown , cell surface molecules such as Q99572 receptor , integrins , CD98 , and macrophage fusion protein are considered to be involved in fusion process . Monocytes of sarcoidosis patients expressed higher levels of Q99572 and had a higher ability to induce MGC than those of healthy controls . Effective agents for sarcoidosis such as tranilast , alloprinol , and captopril inhibited in vitro MGC formation , suggesting their therapeutic effects through the direct effects on monocytes . Thus , an in vitro MGC formation model would be a useful tool to understand the relevance of MGC in granulomatous disorders . A negative feedback loop mediated by P40763 limits human Th17 responses . The transcription factor P40763 is critically required for the differentiation of Th17 cells , a T cell subset involved in various chronic inflammatory diseases . In this article , we report that P40763 also drives a negative-feedback loop that limits the formation of Q16552 -producing T cells within a memory population . By activating human memory P01730 (+)CD45RO(+) T cells at a high density ( HiD ) or a low density ( LoD ) in the presence of the pro-Th17 cytokines IL-1β , IL-23 , and TGF-β , we observed that the numbers of Th17 cells were significantly higher under LoD conditions . Assessment of P40763 phosphorylation revealed a more rapid and stronger P40763 activation in HiD cells than in LoD cells . Transient inhibition of active P40763 in HiD cultures significantly enhanced Th17 cell numbers . Expression of the P40763 -regulated ectonucleotidase P49961 , which catalyzes DB00171 hydrolysis , was higher in HiD , than in LoD , cell cultures . Interestingly , inhibition of P49961 ectonucleotidase activity enhanced Th17 responses under HiD conditions . Conversely , blocking the DB00171 receptor Q99572 reduced Th17 responses in LoD cultures . These data suggest that P40763 negatively regulates Th17 cells by limiting the availability of DB00171 . This negative-feedback loop may provide a safety mechanism to limit tissue damage by Th17 cells during chronic inflammation . Furthermore , our results have relevance for the design of novel immunotherapeutics that target the P40763 -signaling pathway , because inhibition of this pathway may enhance , rather than suppress , memory Th17 responses . P08473 inhibitor suppresses the early phase of atrial electrical remodeling in a canine rapid atrial pacing model . INTRODUCTION : We examined the acute effects of neutral endopeptidase inhibitor on the hemodynamics and electrical properties of dogs subjected to rapid atrial pacing . METHODS : Ten beagle dogs were used and divided into two groups with and without candoxatril , a neutral endopeptidase inhibitor preadministration . Before and after the 6 hours rapid atrial pacing from the right atrial appendage , the hemodynamics , atrial effective refractory period , and monophasic action potential duration of the right atrial appendage were measured and blood samples were collected . Atrial tissue was also excised after the experiment . RESULTS : DB00616 significantly increased plasma P01160 levels ( Control : 88.4 +/- 50.25 vs. DB00616 : 197.1 +/- 32.09 pg/ml , p = 0.004 ) and prevented reductions in atrial effective refractory period and monophasic action potential duration . We further demonstrated that the treated animals exhibited significantly higher levels of atrial tissue cyclic GMP ( Control : 28.1 +/- 1.60 fmol/mg vs. DB00616 : 44.5 +/- 12.28 fmol/mg , p = 0.034 ) as well as that of plasma cyclic GMP ( Control : 32 +/- 5.5 vs. DB00616 : 42 +/- 7.1 pg/ml , p = 0.028 ) . CONCLUSION : DB00616 suppressed the shortening of atrial effective refractory period and monophasic action potential duration in the rapid atrial pacing model . As plasma P01160 and the atrial tissue levels of cyclic GMP were higher in the DB00616 group than the control , this effect was considered to appear through the reduction of calcium overload caused by P01160 and cyclic GMP . 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 . Phospholipase A₂ activities in skin physiology and pathology . Skin inflammatory diseases are most commonly treated with corticosteroids , especially topical preparations , benefitting from high potency and unparalleled formulation flexibility . However , these benefits are limited due to side effects , especially under long-term use . Non-steroidal anti-inflammatory drugs ( NSAIDs ) which block the P36551 pathways have been used as safer alternatives to corticosteroids , and much effort and resources have been invested in developing P36551 inhibitors . However , synthetic NSAIDs are less potent than steroids , have limited formulation flexibility and have their own safety issues , thereby yielding unsatisfactory results , with some high-profile drugs ( e.g. , the P35354 inhibitors Vioxx , DB00482 ) being withdrawn from the market due to safety concerns . The potency and safety challenges of NSAIDs are related to inter-eicosanoid dynamics , pertaining to their pro-versus anti-inflammatory action , homeostatic functions and tissue-specific activities . Instead , the upstream control of phospholipase A2 ( P04054 ) enzymatic activity , which hydrolyzes cell membrane phospholipids to initiate the eicosanoid production , has been considered for inhibiting eicosanoid activation while maintaining the intricate balance needed for their homeostatic functions . Yet , PLA(2) inhibitors have hardly been tested for treating skin inflammatory/allergic conditions . In this article we review the involvement of PLA(2)s in skin physiology and pathology , and discuss the prospect of PLA(2) inhibition for the treatment of dermatological diseases . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . 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 . Action of rolipram on specific DB05876 DB02527 phosphodiesterase isoforms and on the phosphorylation of DB02527 -response-element-binding protein ( CREB ) and p38 mitogen-activated protein ( Q96HU1 ) kinase in U937 monocytic cells . U937 monocytic cells are shown here to express a range of DB05876 , DB02527 -specific phosphodiesterase ( PDE ) isoenzymes : the long isoenzymes , PDE4A4 , PDE4D5 and PDE4D3 , plus the short isoenzyme , PDE4B2 . These isoenzymes provide around 76 % of the total DB02527 PDE activity of U937 cells . The specific activities of the total P27815 , Q07343 and Q08499 activities were 0.63+/-0.09 , 8.8+/-0.2 and 34.4+/-2.9 pmol/min per mg of protein respectively . The DB05876 selective inhibitor , rolipram , inhibited immunopurified Q07343 and Q08499 activities similarly , with IC(50) values of approx . 130 nM and 240 nM respectively . In contrast , rolipram inhibited immunopurified P27815 activity with a dramatically lower IC(50) value of around 3 nM . DB01954 increased phosphorylation of DB02527 -response-element-binding protein ( CREB ) in U937 cells in a dose-dependent fashion , which implied the presence of both high affinity ( IC(50) value approx. 1 nM ) and low affinity ( IC(50) value approx. 120 nM ) components . DB01954 dose-dependently inhibited the interferon-gamma ( P01579 ) -stimulated phosphorylation of p38 mitogen-activated protein ( Q96HU1 ) kinase in a simple monotonic fashion with an IC(50) value of approx . 290 nM . On this basis , it is suggested that rolipram inhibition of PDE4A4 is involved in regulating CREB phosphorylation but not P01579 -stimulated p38 Q96HU1 kinase phosphorylation . PDE4A4 was also selectively activated by challenge of U937 cells with either bacterial lipopolysaccharide ( LPS ) or P01579 through a process which was attenuated by both wortmannin and rapamycin . It is proposed that the PDE4A4 isoform is involved in compartmentalized DB02527 signalling responses in U937 monocytes . 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 .

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interacts_with DB00820?

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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 . Q9Y6W8 -induced B7h shedding on B cells is inhibited by Q9NYK1 /8 and Q9NR96 . We report in this study that B7h , the ligand for the Q9Y6W8 costimulatory receptor , is rapidly shed from mouse B cells following either Q9Y6W8 binding or P11274 engagement . Shedding occurs through proteolytic cleavage that releases the extracellular Q9Y6W8 -binding region of B7h . Prior exposure of B7h-expressing APCs to Q9Y6W8 -expressing cells inhibits their subsequent ability to costimulate P01579 and P05112 production from P01730 + T cells . Shedding is regulated as Q9NYK1 /8 and Q9NR96 ligands inhibit B7h shedding . A shedding-resistant B7h mutant elicits greater costimulation of P01579 production from P01730 + T cells than does wild-type B7h . These data define shedding of B7h as a novel mechanism for controlling costimulatory signaling by P33681 - P10747 family members that is regulated on B cells by TLR signaling . DB00024 -receptor-mediated diseases : a paradigm for receptor autoimmunity . Autoantibodies to the thyrotropin receptor ( P16473 ) can act as thyrotropin agonists or antagonists , or can cause thyroid hypertrophy . Neither the autoantibody-binding sites on the P16473 nor the intracellular mechanisms by which the autoantibodies mediate their diverse functional effects are completely understood . This article reviews how cloning of the P16473 has contributed to our understanding of its structure and function , and has allowed induction of experimental autoimmunity to the P16473 . Antihyperglycemic Activity of Houttuynia cordata Thunb. in DB00428 -Induced Diabetic Rats . Present study is an attempt to investigate plausible mechanism involved behind antidiabetic activity of standardized Houttuynia cordata Thunb. extract in streptozotocin-induced diabetic rats . The plant is used as a medicinal salad for lowering blood sugar level in North-Eastern parts of India . Oral administration of extract at 200 and 400 mg/kg dose level daily for 21 days showed a significant ( P < 0.05 ) decrease in fasting plasma glucose and also elevated insulin level in streptozotocin-induced diabetic rats . It also significantly reversed all the alterations in biochemical parameters , that is , total lipid profile , blood urea , creatinine , protein , and antioxidant enzymes in liver , pancreas , and adipose tissue of diabetic rats . Furthermore , we have demonstrated that the extract significantly reversed the expression patterns of various glucose homeostatic enzyme genes like P11168 , P14672 , and caspase-3 levels but did not show any significant effect on Q07869 - γ protein expressions . Additionally , the extract positively regulated mitochondrial membrane potential and succinate dehydrogenase ( SDH ) activity in diabetic rats . The findings justified the antidiabetic effect of H. cordata which is attributed to an upregulation of P14672 and potential antioxidant activity , which may play beneficial role in resolving complication associated with diabetes . 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 . 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 . P21554 cannabinoid receptor deficiency promotes cardiac remodeling induced by pressure overload in mice . BACKGROUND : The endocannabinoid system is known to play a role in regulating myocardial contractility , but the influence of cannabinoid receptor 1 ( P21554 ) deficiency on chronic heart failure ( CHF ) remains unclear . In this study we attempted to investigate the effect of P21554 deficiency on CHF induced by pressure overload and the possible mechanisms involved . METHODS AND RESULTS : A CHF model was created by transverse aortic constriction ( TAC ) in both P21554 knockout mice and wild-type mice . P21554 knockout mice showed a marked increase of mortality due to CHF from 4 to 8 weeks after TAC ( p=0.021 ) . Five weeks after TAC , in contrast to wild-type mice , P21554 knockout mice had a higher left ventricular ( LV ) end-diastolic pressure , lower rate of LV pressure change ( ± dp/dt max ) , lower LV contractility index , and a larger heart weight to body weight ratio and lung weight to body weight ratio compared with wild-type mice ( all p < 0.05-0.001 ) . Phosphorylation of the epidermal growth factor receptor ( P00533 ) and mitogen-activated protein kinases ( O75791 and P29323 ) was higher in P21554 knockout mice than that in wild-type mice . In cultured neonatal rat cardiomyocytes , a P21554 agonist reduced DB02527 production stimulated by isoproterenol or forskolin , and suppressed phosphorylation of the P00533 , O75791 , and P29323 , while the inhibitory effect of a P21554 agonist on P00533 phosphorylation was abrogated by P21554 knockdown . CONCLUSION : These findings indicate that cannabinoid receptor 1 inactivation promotes cardiac remodeling by enhancing the activity of the epidermal growth factor receptor and mitogen-activated protein kinases . Anti-clastogenic effect of beta-glucan extracted from barley towards chemically induced DNA damage in rodent cells . beta-Glucan ( BG ) was tested in vitro to determine its potential clastogenic and/or anti-clastogenic activity , and attempts were made to elucidate its possible mechanism of action by using combinations with an inhibitor of DNA polymerase . The study was carried out on cells deficient ( CHO-k1 ) and cells proficient ( HTC ) in phases I and II enzymes , and the DNA damage was assessed by the chromosomal aberration assay . BG did not show a clastogenic effect , but was anti-clastogenic in both cell lines used , and at all concentrations tested ( 2.5 , 5 and 10 microg/mL ) in combination with damage inducing agents ( methylmethane sulfonate in cell line CHO-k1 , and methylmethane sulfonate or 2-aminoanthracene in cell line HTC ) . BG also showed a protective effect in the presence of a P06746 inhibitor ( cytosine arabinoside-3-phosphate , DB00987 ) , demonstrating that BG does not act through an anti-mutagenic mechanism of action involving P06746 . Neuroprotective and abstinence-promoting effects of acamprosate : elucidating the mechanism of action . DB00659 is an abstinence-promoting drug widely used in the treatment of alcohol dependence but which has a mechanism of action that has remained obscure for many years . Recently , evidence has emerged that this drug may interact with excitatory glutamatergic neurotransmission in general and as an antagonist of the metabotropic glutamate receptor subtype 5 ( P41594 ) in particular . These findings provide , for the first time , a satisfactory , unifying hypothesis that can bring together and explain the diverse neurochemical effects of acamprosate . DB00142 is involved in several aspects of alcohol dependence and withdrawal , many of which can be modified by acamprosate . For example , during chronic exposure to alcohol , the glutamatergic system becomes upregulated , leaving the brain exposed to excessive glutamatergic activity when alcohol is abruptly withdrawn . The surge in glutamic acid release that occurs following alcohol withdrawal can be attenuated by acamprosate . The elevated extracellular levels of glutamic acid observed in withdrawal , together with supersensitivity of DB01221 receptors , may expose vulnerable neurons to excitotoxicity , possibly contributing to the neuronal loss sometimes observed in chronic alcohol dependence . In vitro studies suggest that the excitotoxicity produced by ethanol can effectively be blocked by acamprosate . Moreover , glutamatergic neurotransmission plays an important role in the acquisition of cue-elicited drinking behaviours , which again can be modulated by acamprosate . In conclusion , the glutamatergic hypothesis of the mechanism of action of acamprosate helps explain many of its effects in human alcohol dependence and points the way to potential new activities , such as neuroprotection , that merit exploration in the clinic . 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 . 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 . P60568 suppression by 2-arachidonyl glycerol is mediated through peroxisome proliferator-activated receptor gamma independently of cannabinoid receptors 1 and 2 . 2-Arachidonyl glycerol ( 2-AG ) is an endogenous arachidonic acid derivative that binds cannabinoid receptors P21554 and CB2 and is hence termed an endocannabinoid . 2-AG also modulates a variety of immunological responses , including expression of the autocrine/paracrine T cell growth factor interleukin ( IL ) -2 . The objective of the present studies was to determine the mechanism responsible for P60568 suppression by 2-AG . Because of the labile properties of 2-AG , 2-AG ether , a nonhydrolyzable analog of 2-AG , was also used . Both 2-AG and 2-AG ether suppressed P60568 expression independently of P21554 and CB2 , as demonstrated in leukocytes derived from P21554 /CB2-null mice . Moreover , we demonstrated that both 2-AG and 2-AG ether treatment activated peroxisome proliferator-activated receptor gamma ( PPARgamma ) , as evidenced by forced differentiation of 3T3- Q9NUQ9 cells into adipocytes , induction of aP2 mRNA levels , and activation of a PPARgamma-specific luciferase reporter in transiently transfected 3T3- Q9NUQ9 cells . Consequently , the putative role of PPARgamma in P60568 suppression by 2-AG and 2-AG ether was examined in Jurkat T cells . Concordant with PPARgamma involvement , the PPARgamma-specific antagonist 2-chloro-5-nitro-N-(4-pyridyl)-benzamide ( T0070907 ) blocked 2-AG- and 2-AG ether-mediated P60568 suppression . Likewise , 2-AG suppressed the transcriptional activity of two transcription factors crucial for P60568 expression , nuclear factor of activated T cells and nuclear factor kappaB , in the absence but not in the presence of T0070907 . 2-AG treatment also induced PPARgamma binding to a Q07869 response element in activated Jurkat T cells . Together , the aforementioned studies identify PPARgamma as a novel intracellular target of 2-AG , which mediates the suppression of P60568 by 2-AG in a manner that is independent of P21554 and/or CB2 . 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 . Increased expression of pro-angiogenic factors and vascularization in thyroid hyperfunctioning adenomas with and without DB00024 receptor activating mutations . Autonomously functioning thyroid nodules ( AFTN ) are known to receive an increased blood influx necessary to sustain their high rate of growth and hormone production . Here , we investigated the expression of hematic and lymphatic vases in a series of 20 AFTN compared with the contralateral non-tumor tissues of the same patients , and the transcript levels of proteins involved in the control of vascular proliferation , including the vascular endothelial growth factor ( P15692 ) and platelet-derived growth factors ( PDGF ) and their receptors and the endothelial nitric oxide synthase ( P29474 ) . In parallel , the expression of the differentiation markers sodium/iodide symporter ( Q92911 ) , thyroperoxidase ( P07202 ) , thyroglobulin ( Tg ) , and DB00024 receptor ( P16473 ) was also investigated . The data were further analyzed comparing subgroups of tumors with or without mutations in the P16473 gene . Analysis by means of CD31 and D2-40 immunostaining showed in AFTN an increased number of hematic , but not lymphatic , vessels in parallel with an enhanced proliferation rate shown by increased Ki67 staining . Quantitative RT-PCR analysis revealed an increase of P15692 , P17948 and 2 , PDGF-A , PDGF-B , and P29474 expression in tumor versus normal tissues . Also , higher transcript levels of Q92911 , P07202 , and Tg were detected . Comparison of the two subgroups of samples revealed only few differences in the expression of the genes examined . In conclusion , these data demonstrate an increased expression of angiogenesis-related factors associated with an enhanced proliferation of hematic , but not lymphatic , vessels in AFTNs . In this context , the presence of P16473 mutations may only slightly influence the expression of pro-angiogenic growth factors . Inhibition of mammalian DNA polymerases by recombinant alpha-interferon and gamma-interferon . Interferons ( IFNs ) have been shown to suppress the growth of both normal and malignant cells . We examined the effect of gene-cloned IFN-alpha and P01579 on the in vitro activities of human , calf , or rat DNA polymerases . IFN-alpha strongly inhibited the reactions of DNA polymerase alpha and beta at apparent Ki values of 1.25 and 0.35 x 10(5) antiviral units/ml , respectively , but inhibited DNA polymerase gamma only slightly . P01579 inhibited the reaction of DNA polymerase alpha more strongly ( Ki , 1.2 x 10(4) units/ml ) than IFN-alpha , but not that of P06746 . On the other hand , neither IFN-alpha nor P01579 inhibited the reactions of DNA polymerase I from Escherichia coli , Klenow fragment , T-4 DNA polymerase , and RNA polymerase . The fact that Ki values for IFN-alpha of DNA polymerase from calf thymus , human leukemic cells , and rat liver were similar suggests the absence of species specificity among animals with regard to the inhibition of DNA polymerases by IFNs . These results indicate that DNA polymerase may be one of the targets of the action of IFNs . Q9Y6W8 ligation recruits the p50alpha PI3K regulatory subunit to the immunological synapse . Q9Y6W8 ligation in concert with TCR stimulation results in strong PI3K activation in T lymphocytes . The Q9Y6W8 cytoplasmic tail contains an YMFM motif that binds the p85alpha subunit of class IA PI3K , similar to the YMNM motif of P10747 , suggesting a redundant function of the two receptors in PI3K signaling . However , Q9Y6W8 costimulation shows greater PI3K activity than P10747 in T cells . We show in this report that Q9Y6W8 expression in activated T cells triggers the participation of p50alpha , one of the regulatory subunits of class IA PI3Ks . Using different T- P25054 cell conjugate systems , we report that p50alpha accumulates at the immunological synapse in activated but not in resting T cells . Our results demonstrate that Q9Y6W8 membrane expression is involved in this process and that p50alpha plasma membrane accumulation requires a functional YMFM Src homology 2 domain-binding motif in Q9Y6W8 . We also show that Q9Y6W8 triggering with its ligand , O75144 , induces the recruitment of p50alpha at the synapse of T cell/ P25054 conjugates . In association with the p110 catalytic subunit , p50alpha is known to carry a stronger lipid kinase activity compared with p85alpha . Accordingly , we observed that Q9Y6W8 engagement results in a stronger activation of PI3K . Together , these findings provide evidence that p50alpha is likely a determining factor in Q9Y6W8 -mediated PI3K activity in T cells . These results also suggest that a differential recruitment and activity of class IA PI3K subunits represents a novel mechanism in the control of PI3K signaling by costimulatory molecules . 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 . 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 . Endotoxemia and sepsis mortality reduction by non-anticoagulant activated protein C . DB00055 ( P25054 ) reduces mortality of severe sepsis patients but increases the risk of serious bleeding . P25054 exerts anticoagulant activity by proteolysis of factors Va/VIIIa . P25054 also exerts antiinflammatory and antiapoptotic effects and stabilizes endothelial barrier function by P25054 -initiated cell signaling that requires two receptors , endothelial cell protein C receptor ( Q9UNN8 ) and protease-activated receptor 1 ( PAR1 ) . The relative importance of P25054 's various activities for efficacy in sepsis is unknown . We used protein engineering of mouse P25054 and genetically altered mice to clarify mechanisms for the efficacy of P25054 in mouse sepsis models . Mortality reduction in LPS-induced endotoxemia required the enzymatic active site of P25054 , Q9UNN8 , and P25116 , highlighting a key role for P25054 's cytoprotective actions . A recombinant P25054 variant with normal signaling but < 10 % anticoagulant activity ( 5A- P25054 ) was as effective as wild-type P25054 in reducing mortality after LPS challenge , and enhanced the survival of mice subjected to peritonitis induced by gram-positive or -negative bacteria or to polymicrobial peritoneal sepsis triggered by colon ascendens stent implantation . Thus , P25054 's efficacy in severe sepsis is predominantly based on Q9UNN8 - and PAR1-dependent cell signaling , and P25054 variants with normal cell signaling but reduced anticoagulant activities retain efficacy while reducing the risk of bleeding . 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 . Unsaturated side chain beta-11-hydroxyhexahydrocannabinol analogs . The cannabinoid side chain is a key pharmacophore in the interaction of cannabinoids with their receptors ( P21554 and CB2 ) . To study the stereochemical requirements of the side chain , we synthesized a series of cannabinoids in which rotation around the C1'- P06681 ' bond is blocked . The key steps in the synthesis were the cuprate addition of a substituted resorcinol to (+)-apoverbenone , the TMSOTf-mediated formation of the dihydropyran ring , and the stereospecific introduction of the beta-11-hydroxymethyl group . All the analogs tested showed nanomolar affinity for the receptors , the cis-hept-1-ene side chain having the highest affinity for P21554 ( Ki = 0.89 nM ) and showing the widest separation between P21554 and CB2 affinities . The parent n-heptyl-beta-11-hydroxyhexahydrocannabinol was the least potent binding to P21554 ( Ki = 8.9 nM ) and had the lowest selectivity between P21554 and CB2 . P06850 drives anandamide hydrolysis in the amygdala to promote anxiety . P06850 ( P06850 ) is a central integrator in the brain of endocrine and behavioral stress responses , whereas activation of the endocannabinoid P21554 receptor suppresses these responses . Although these systems regulate overlapping functions , few studies have investigated whether these systems interact . Here we demonstrate a novel mechanism of P06850 -induced anxiety that relies on modulation of endocannabinoids . Specifically , we found that P06850 , through activation of the P06850 receptor type 1 ( P34998 ) , evokes a rapid induction of the enzyme fatty acid amide hydrolase ( FAAH ) , which causes a reduction in the endocannabinoid anandamide ( AEA ) , within the amygdala . Similarly , the ability of acute stress to modulate amygdala FAAH and AEA in both rats and mice is also mediated through P34998 activation . This interaction occurs specifically in amygdala pyramidal neurons and represents a novel mechanism of endocannabinoid- P06850 interactions in regulating amygdala output . Functionally , we found that P06850 signaling in the amygdala promotes an anxious phenotype that is prevented by FAAH inhibition . Together , this work suggests that rapid reductions in amygdala AEA signaling following stress may prime the amygdala and facilitate the generation of downstream stress-linked behaviors . Given that endocannabinoid signaling is thought to exert " tonic " regulation on stress and anxiety responses , these data suggest that P06850 signaling coordinates a disruption of tonic AEA activity to promote a state of anxiety , which in turn may represent an endogenous mechanism by which stress enhances anxiety . These data suggest that FAAH inhibitors may represent a novel class of anxiolytics that specifically target stress-induced anxiety .

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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 . Targeting reduction of proteinuria in glomerulonephritis : Maximizing the antifibrotic effect of valsartan by protecting podocytes . Although angiotensin ( Ang ) II blockade has become a standard antifibrotic therapy in kidney disease , the therapeutic efficacy of Ang II blockade is yet to be optimized . Considering the prognostic impact of proteinuria reduction , we hypothesized that titration of Ang II blockade for optimal anti-proteinuric effect would improve renoprotection . One day after induction of Thy 1.1 glomeruonephritis , rats were treated with increasing doses of the Ang II receptor blocker valsartan in drinking water . Six days after disease induction , the therapeutic effect on proteinuria , podocyte injury and glomerular fibrosis was evaluated . Increasing doses of valsartan resulted in increasing reduction of proteinuria . The maximally effective dose of valsartan was determined to be 1000 mg/l , which reduced proteinuria by 80 % and maximally reduced glomerular matrix expansion , fibronectin , collagen I and collagen III staining and glomerular mRNAs for TGFß1 , P05121 , FN and collagen I . Notably , valsartan given at this dose prevented podocyte dysfunction by preserving expression of podocin and nephrin and the counter-regulating molecule P33681 -1 that is involved in podocyte injury . These results support the hypothesis that higher doses of valsartan are required to optimize proteinuria reduction and glomerulosclerosis amelioration . Further , the optimal dose of valsartan also provides an additional therapeutic effect by preventing podocyte dysfunction . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . Growth regulatory properties of endothelins . Endothelins are produced by endothelial and epithelial cells , macrophages , fibroblasts , and many other types of cells . Their receptors are present in numerous cells , including smooth muscle cells , myocytes , and fibroblasts . Evidence now suggests that the three isoforms of endothelins ( ET-1 and the other two related isopeptides , P20800 and P14138 ) regulate growth in several of these cells . P05305 influences DNA synthesis , the expression of protooncogenes , cell proliferation , and hypertrophy . The participation of ET in mitogenesis involves activation of multiple transduction pathways , such as the production of second messengers , the release of intracellular pools of calcium , and influx of extracellular calcium . Moreover , ET-1 acts in synergism with various factors , such as P01133 , PDGF , P09038 , TGFs , insulin , etc. , to potentiate cellular transformation or replication . Several of these factors may in turn stimulate the synthesis and/or the release of endothelins . The production and release of endothelins are also increased in acute and chronic pathological processes , e.g. , atherosclerosis , postangioplastic restenosis , hypertension , and carcinogenesis . It is postulated that endothelins act in a paracrine/autocrine manner in growth regulation and play an important role mediating vascular remodeling in some cardiovascular diseases . The present review analyses the implication of endothelins ( ET-1 , -2 , and -3 ) in physiopathology related to their growth regulatory properties . 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 . 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 . 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 . 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 . [ Endothelial progenitor cells related gene expression changes before and early after revascularization in patients with acute myocardial infarction ] . OBJECTIVE : The purpose of this study was to observe the endothelial progenitor cells ( EPCs ) related gene expression changes before and early after revascularization in patients with acute myocardial infarction . METHODS : Peripheral blood samples were taken from patients with acute anterior myocardial infarction 6 hours and 7 days after P05154 and stenting . Mononuclear cells ( MNCs ) were isolated by Ficoll-density centrifugation and cultured in M-199 medium . After 14 days culture , attaching cells incorporated DiI-acetylated low-density lipoprotein ( EPCs ) were collected and RNA was isolated by Trizol for microarray analysis on 24 genes associated with permissibility/vessel tone ( angiotensin system : P12821 , AGTR-1 , AGTR-2 ; NO system : P29474 ; prostacyclin system : P35354 ; endothelin system : ET-1 , P25101 , ETB ; superoxide anions system : SOD-1 ) , angiogenesis ( adhesion molecule : P33151 ; growth factors and receptors : P17948 , P35968 , P15692 ) and endothelial cell activation ( adhesion molecules expression : P05362 , P13598 , P32942 , P16284 , E-Selectin , L-Selection , P19320 ; change phenotype from antithrombotic to prothrombotic : tPA , uPA , P05121 , P04275 ) . P35968 , P16284 and P33151 positive cells were identified by flow cytometry . RESULT : Eight gene expressions ( AGTR-1 , AGTR-2 , P35354 , P29474 , ET-1 , P25101 , P15692 ) were significantly downregulated 7 days post P05154 compared to pre- P05154 ( P < 0.05 ) . Flow cytometry results showed that P35968 positive cells were also significantly reduced post P05154 than that of before P05154 ( P < 0.05 ) . CONCLUSION : P05154 down-regulated endothelial progenitor cells related gene expressions in patients with acute myocardial infarction . DB04901 ( IDEC ) . IDEC is developing a PRIMATIZED-anti- P33681 antibody ( DB04901 ) for the treatment of autoimmune and inflammatory diseases , such as psoriasis and rheumatoid arthritis . It is currently undergoing phase II trials in patients with psoriasis [ 395813 ] . A randomized , blind , placebo-controlled , multiple-dose phase II study was initiated in January 2001 to evaluate the potential clinical activity and safety of DB04901 in patients with moderate-to-severe psoriasis [ 395813 ] . The antibody targets the P33681 antigen on the surface of antigen-presenting cells that normally interact with T-cells to initiate an immune response . Antibodies directed at P33681 may be useful in preventing unwanted immune responses in autoimmune diseases such as systemic lupus erythematosus , idiopathic thrombocytopenic purpura as well as transplant rejection [ 178382 ] , [ 178929 ] . PRIMATIZED antibodies , genetically engineered from cynomolgus macaque monkey and human components , are structurally indistinguishable from human antibodies . They may , therefore , be less likely to cause adverse reactions in humans , making them potentially suited for long-term , chronic treatment [ 244805 ] . IDEC has signed an antibody humanization patent licensing agreement with Protein Design Labs [ 240591 ] . IDEC is also collaborating with Mitsubishi-Tokyo ( formerly Mitsubishi Kasei ) on the development of this antibody [ 178382 ] . 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 . G- P04141 increases secretion of urokinase-type plasminogen activator by human lung cancer cells . We reported previously that granulocyte colony-stimulating factor ( DB00099 ) can promote the invasion of human lung cancer cell lines in vitro . However , the exact mechanism of its stimulatory effect on invasion remains to be elucidated . In the present study we mainly focused our attention on the components of the plasminogen activation system in human lung cancer cell lines , because of the central role that plasminogen activators play in regulating extracellular proteolysis . We showed that G- P04141 induced a dose-dependent increase in the urokinase-type plasminogen activator ( uPA ) activity in the conditioned medium of a PC-9 lung cancer cell line . When the amounts of uPA activity were quantitated by densitometry , we found that even at a concentration of 0.01 microg/ml , G- P04141 had a stimulatory effect on the uPA release , while high concentrations caused a 3.6-fold increase at a maximum concentration of 1 microg/ml . A Western blot analysis of the conditioned medium confirmed the findings observed in a zymographic analysis . The observed increase in uPA protein was paralleled by a significant increase in the uPA mRNA levels after treatment with DB00099 . However , our experiments failed to identify any alteration in the plasminogen activator inhibitor ( P05121 ) secretion caused by DB00099 . In addition , we also found the expression of Q99062 by PC-9 cells , suggesting the possible pathway activated by DB00099 . Impact of high salt independent of blood pressure on PRMT/ DB01686 /DDAH pathway in the aorta of Dahl salt-sensitive rats . Endothelial dysfunction participates in the development and progression of salt-sensitive hypertension . DB01686 ( DB01686 ) is an endogenous inhibitor of nitric oxide synthase ( NOS ) . The objectives of this study were to investigate the impact of a high salt diet on the PRMT/ DB01686 /DDAH ( protein arginine methyltransferases ; dimethylarginine dimethylaminohydrolase ) pathway in Dahl salt-sensitive ( DS ) rats and SS-13BN consomic ( DR ) rats , and to explore the mechanisms that regulate DB01686 metabolism independent of blood pressure reduction . Plasma levels of nitric oxide ( NO ) in DS rats given a high salt diet and subjected to intragastric administration of hydralazine ( SH + O95071 group ) were lower than those given a normal salt diet ( SN group ) . There were significant decreases in expression and activity of dimethylarginine dimethylaminohydrolase ( DDAH ) and endothelial NO synthase ( P29474 ) in DS rats given a high diet ( SH group ) in comparison to the SN group . The activity of DDAH and expression of P29474 in the SH + O95071 group decreased more significantly than SN group . The mRNA expression of O94760 and O95865 were lowest in the SH group . The results suggest that salt , independent of blood pressure , can affect the PRMT-1/ DB01686 /DDAH system to a certain degree and lead to endothelial dysfunction in Dahl salt-sensitive rats . DB08901 suppresses the development of myeloid and lymphoid malignancies associated with P11362 abnormalities . Myeloid and lymphoid malignancies associated with fibroblast growth factor receptor-1 ( P11362 ) abnormalities are characterized by constitutively activated P11362 kinase and rapid transformation to acute myeloid leukemia and lymphoblastic lymphoma . Molecular targeted therapies have not been widely used for stem cell leukemia/lymphoma ( SCLL ) . DB08901 ( DB08901 ) , which potently inhibits native and mutant P11274 - P00519 , also targets the FGFR family . Using murine BaF3 cells , stably transformed with six different P11362 fusion genes , as well as human KG1 cells expressing activated chimeric P11362 and five newly established murine SCLL cell lines , we show that ponatinib ( < 50 nM ) can effectively inhibit phosphoactivation of the fusion kinases and their downstream effectors , such as PLCγ , Stat5 and Src . DB08901 also significantly extended survival of mice transplanted with different SCLL cell lines . DB08901 administered at 30 mg/kg daily also significantly delayed , or even prevented , tumorigenesis of KG1 cells in xenotransplanted mice . Furthermore , we demonstrate that ponatinib specifically inhibits cell growth and clonogenicity of normal human P28906 + progenitor cells transformed by chimeric P11362 fusion kinases . Overall , our data provide convincing evidence to suggest that pharmacologic inhibition of P11362 fusion kinases with ponatinib is likely to be beneficial for patients with SCLL and perhaps for other human disorders associated with dysregulated P11362 activity . A review of sitaxsentan sodium in patients with pulmonary arterial hypertension . Pulmonary arterial hypertension ( PAH ) is a life threatening , progressive condition which eventually leads to fatal right heart failure . P05305 ( ET-1 ) , a potent vasoconstrictor peptide , is increased in the pulmonary arteries of patients with pulmonary hypertension . P05305 acts through the stimulation of 2 subtypes of receptors ( endothelin receptor subtypes A [ ET(A) ] and B [ ET(B) ] ) . In PAH patients , ETRAs block the deleterious vasoconstrictor effects of ET-1 , and P25101 treatment in PAH patients has been shown to be safe and efficacious . DB06268 is an orally active , highly ET(A) selective P25101 that , in clinical trials , has demonstrated improvements in exercise capacity , functional class and hemodynamics in PAH patients . DB06268 has been shown to be safe , well tolerated , and associated with a lower incidence of liver toxicity than other approved ETRAs . DB09222 binding potentiates P09038 but not P15692 induced expression of u-PA , u-PAR , and P05121 in endothelial cells . Endothelial cell responses at sites of injury occur in a fibrin matrix and are regulated by growth factors including those of the FGF and P15692 families . The pericellular proteolytic balance is important in these responses , and P09038 and P15692 up-regulate endothelial cell u-PA , u-PAR and P05121 . Because both P15692 and P09038 bind to fibrinogen , we have examined the capacity of fibrinogen to modulate the up-regulation of these proteins by P09038 and P15692 . Confluent cultures of endothelial cells were exposed to P09038 , P15692 , and fibrinogen or to combinations of growth factors with fibrinogen . Changes in mRNA levels of u-PA , u-PAR and P05121 were measured by Northern blot . P09038 increased u-PA , u-PAR , and P05121 mRNA , but there was a significantly greater induction when fibrinogen was added to P09038 at all concentrations . The potentiation by fibrinogen was particularly evident at an P09038 concentration of 0.1 ng mL(-1) , which resulted in non-significant change in transcript levels by itself , but significantly increased up to 2.6-fold with fibrinogen . P15692 also increased endothelial cell expression of u-PA , u-PAR and P05121 , but this effect was not potentiated by fibrinogen . Addition of LM609 , a monoclonal antibody to alphaVbeta3 , significantly inhibited induction of u-PA mRNA and activity by fibrinogen-bound P09038 compared to P09038 . A monoclonal antibody to P11362 also inhibited u-PA mRNA expression induced by fibrinogen-bound P09038 . We conclude that fibrinogen increases the capacity of P09038 , but not of P15692 , to up-regulate u-PA , u-PAR , and P05121 in endothelial cells and that fibrinogen-bound P09038 requires alphaVbeta3 binding to up-regulate endothelial cell u-PA . 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 . A novel pathway involving progesterone receptor , endothelin-2 , and endothelin receptor B controls ovulation in mice . The steroid hormone progesterone ( P ) plays a pivotal role during ovulation . Mice lacking P receptor ( Pgr ) gene fail to ovulate due to a defect in follicular rupture . The P receptor ( P06401 ) -regulated pathways that modulate ovulation , however , remain poorly understood . To identify these pathways , we performed gene expression profiling using ovaries from mice subjected to gonadotropin-induced superovulation in the presence and in the absence of DB05366 , a synthetic P06401 antagonist . Prominent among the genes that were down-regulated in response to DB05366 was endothelin ( ET ) -2 , a potent vasoactive molecule . P20800 mRNA was transiently induced in mural granulosa cells of the preovulatory follicles immediately preceding ovulation . This induction was absent in the ovaries of P06401 null mice , indicating a critical role of this receptor in P20800 expression . To investigate the functional role of P20800 during ovulation , we employed selective antagonists of endothelin receptors , ETR-A and ETR-B . Mice treated with an ETR-B antagonist exhibited a dramatic ( > 85 % ) decline in the number of released oocytes . Strong expression of ETR-B was observed in the mural and cumulus granulosa cells of the preovulatory follicles as well as in the capillaries lining the inner border of the theca interna . We also identified Q13237 , a previously reported P06401 -regulated gene , as a downstream target of P20800 during ovulation . Collectively , our studies uncovered a unique pathway in which P20800 , produced by P06401 in mural granulosa cells , acts in a paracrine or autocrine manner on multiple cell types within the preovulatory follicle to control the final events leading to its rupture . DB08626 -induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer 's disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 and cortical Abeta40 that were 147 and 142 % of the control group , respectively . Results for Abeta42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD . Association study between Alzheimer 's disease and genes involved in Abeta biosynthesis , aggregation and degradation : suggestive results with P56817 . BACKGROUND : Amyloid beta-peptide ( Abeta ) biosynthesis , aggregation and degradation constitute three important steps to consider in the study of pathological mechanisms involved in Alzheimer 's disease ( AD ) . Several proteins have been suggested as involved in each of these processes : proteolytic cleavage of the amyloid precursor protein by the beta-site P05067 cleaving enzyme ( P56817 ) , increased amyloid fibril formation by the activity of the acetylcholinesterase ( P22303 gene ) , and degradation of Abeta aggregates by the plasmin system have been exhaustively documented . METHODS : A case-control design was used to evaluate the possible association between candidate genes involved in these three processes and AD . We analysed three polymorphisms located at the P56817 gene , one polymorphism at the P22303 gene , and two variants located at the tissue plasminogen activator and plasminogen activator inhibitor-1 ( genes TPA and P05121 - 1 , respectively ) , both part of the plasmin system . RESULTS : We found an association between P56817 exon 5 GG genotype and AD ( age-and gender-adjusted odds ratio = 2.14 , P =0.014 ) . Although a similar association was reported previously by Nowotny and collaborators only in subjects carrying the epsilon4-allele of the apolipoprotein E gene ( P02649 ) , we did not detect this effect . However,when we combined our results with those previously reported , a clear increase of the risk to develop AD appeared in subjects carrying both the P56817 exon 5 GG genotype and the P02649 epsilon4-allele ( crude OR = 2.2 , P = 0.004 ) . CONCLUSION : These data suggest a possible genetic relation between P56817 and AD .

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interacts_with DB04905?

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Long-term stability of international reference preparations for thromboplastins . Three certified reference materials for thromboplastins are available from the Community Bureau of Reference ( P11274 ) of the European Commission for calibration of commercial thromboplastins used for control of oral anticoagulant therapy . The long-term stability of these reference materials has been monitored by two independent laboratories , using deep-frozen and lyophilized plasma samples . P00734 times and prothrombin time ratios measured on 19 occasions in the period 1981-86 have been analysed for trend with time . Although significant trends of prothrombin time and ratio ( P less than 0.05 ) were observed , a consistent pattern of trends could not be recognized . The significant trends of prothrombin time and prothrombin time ratio are most probably due to changes in local laboratory conditions . There is no indication that the reference materials have deteriorated since the beginning of the study . It is recommended that long-term stability monitoring of thromboplastins be performed by at least two laboratories simultaneously . Acute pharmacodynamic and antivascular effects of the vascular endothelial growth factor signaling inhibitor DB04849 in Calu-6 human lung tumor xenografts . The vascular endothelial growth factor-A ( P15692 ) signaling pathway , a key stimulant of solid tumor vascularization , is primarily dependent on the activation of the endothelial cell surface receptor P15692 receptor-2 ( P35968 ) . DB04849 is an oral , highly potent small-molecule inhibitor of VEGFR tyrosine kinase activity that inhibits angiogenesis and the growth of human tumor xenografts in vivo . Here , we show pharmacodynamic changes in P35968 phosphorylation induced by DB04849 . In mouse lung tissue , a single dose of DB04849 at 6 mg/kg inhibited P15692 -stimulated P35968 phosphorylation by 87 % at 2 h with significant inhibition ( > or=60 % ) maintained to 24 h . To examine inhibition of P35968 phosphorylation in tumor vasculature by immunohistochemistry , a comprehensive assessment of antibodies to various phosphorylation sites on the receptor was undertaken . Antibodies to the phosphotyrosine epitopes pY1175/1173 and pY1214/1212 were found suitable for this application . Calu-6 human lung tumor xenografts , from mice receiving DB04849 or vehicle treatment ( p.o. , once daily ) , were examined by immunohistochemistry . A significant reduction in tumor vessel staining of phosphorylated P35968 ( pVEGFR-2 ) was evident within 28 h of DB04849 treatment ( 6 mg/kg ) . This effect preceded a significant reduction in tumor microvessel density , which was detectable following 52 h of DB04849 treatment . These data show that DB04849 is a potent inhibitor of P35968 activation in vivo and suggest that DB04849 delivers therapeutic benefit in Calu-6 tumors by targeting vessels dependent on P35968 signaling for survival . In addition , this work highlights the utility of measuring either pY1175/1173 or pY1214/1212 on P35968 as a pharmacodynamic marker of P35968 activation . 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 . 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 . 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 . 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 . The expression of the solute carriers Q14973 and O75051 -1 is regulated by cholesterol in HepG2 cells . Drug disposition and response are greatly determined by the activities of drug-metabolizing enzymes and transporters . While the knowledge in terms of CYP enzymes and efflux ABC transporters ( such as P08183 , P-glycoprotein ) is quite extensive , influx transporters are increasingly being unveiled as key contributors to the process of drug disposition . There is little information on the regulation of these proteins in human cells , especially as regards the effect of endogenous compounds . In this study , we analysed the expression of P08684 and three uptake transporters Q14973 ( Q14973 ) , P46721 / P46721 ( P46721 ) and O75051 -1 ( O15245 ) in HepG2 cells following treatment with cholesterol . While P08684 and P46721 expression was unaffected , cholesterol treatment led to increased levels of Q14973 and O75051 -1 mRNAs . Alterations in the functional characteristics and/or expression levels of drug transporters in the liver may conceivably contribute to the variability in drug oral bioavailability often observed in the clinical settings . 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 . Dissociation between cognitive and motor/motivational deficits in the delayed matching to position test : effects of scopolamine , 8-OH-DPAT and EAA antagonists . The effects of the muscarinic antagonists scopolamine HBr and MeBr , the P08908 agonist 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , and the N-methyl-d-aspartate ( DB01221 ) antagonists MK-801 and CGS-19755 on performance of rats in a delayed matching-to-position task were examined . Pretreatment with scopolamine HBr ( 0.05 and 0.1 mg/kg ) , resulted in a delay-dependent decrease in the percentage of correct responses and discriminability ( log d ) , but had no effect on either the latency to complete trials , or the rate of trial completion during the fixed duration session . DB00747 MeBr ( 0.1 mg/kg ) did not impair percent correct or increase the response latency but did decrease the rate of trial completion . 8-OH-DPAT ( up to 0.3 mg/kg ) , had no effect on percent correct , but did induce a small decrease in discriminability . The impairment in discriminability occurred only at a dose that substantially reduced the rate of trial completion . Both MK-801 ( 0.05 mg/kg ) and CGS 19755 ( 10 mg/kg ) induced a delay-independent impairment in percent correct , discriminability and a reduction in the rate of trial completion without affecting latency . A lower dose of CGS 19755 ( 5.0 mg/kg ) induced a slight impairment in discriminability without significantly affecting the other measures . Taken together , these results demonstrate some dissociation between drug-induced cognitive and motor/motivational deficits in the DMTP test . However , the data question the specificity of putative cognitive impairments reported in many previous studies with the P08908 agonist 8-OH-DPAT . Presynaptic serotonergic inhibition of GABAergic synaptic transmission in mechanically dissociated rat basolateral amygdala neurons . 1. The basolateral amygdala ( P00519 ) nuclei contribute to the process of anxiety . GABAergic transmission is critical in these nuclei and serotonergic inputs from dorsal raphe nuclei also significantly regulate GABA release . In mechanically dissociated rat P00519 neurons , spontaneous miniature inhibitory postsynaptic currents ( mIPSCs ) arising from attached GABAergic presynaptic nerve terminals were recorded with the nystatin-perforated patch method and pharmacological isolation . 2 . 5-HT reversibly reduced the GABAergic mIPSC frequency without affecting the mean amplitude . The serotonergic effect was mimicked by the P08908 specific agonist 8-OH DPAT ( 8-hydroxy-2-(di-n-propylamino)tetralin ) and blocked by the P08908 antagonist spiperone . 3 . The GTP-binding protein inhibitor N-ethylmaleimide removed the serotonergic inhibition of mIPSC frequency . In either K+-free or Ca2+-free external solution , 5-HT could inhibit mIPSC frequency . 4 . High K+ stimulation increased mIPSC frequency and 8-OH DPAT inhibited this increase even in the presence of Cd2+ . 5 . DB02587 , an activator of adenylyl cyclase ( AC ) , significantly increased synaptic GABA release frequency . Pretreatment with forskolin prevented the serotonergic inhibition of mIPSC frequency in both the standard and high K+ external solution . 6 . Ruthenium Red ( RR ) , an agent facilitating the secretory process in a Ca2+-independent manner , increased synaptic GABA release . 5-HT also suppressed RR-facilitated mIPSC frequency . 7 . We conclude that 5-HT inhibits GABAergic mIPSCs by inactivating the AC- DB02527 signal transduction pathway via a G-protein-coupled P08908 receptor and this intracellular pathway directly acts on the GABA-releasing process independent of K+ and Ca2+ channels in the presynaptic nerve terminals . Q92817 P11274 / P00519 kinase regulates nucleotide excision repair ( P55055 ) and resistance to UV radiation . Both clinical and experimental evidence illustrate that P78357 and Q92817 P11274 / P00519 oncogenic tyrosine kinases induce resistance to DNA damage and confer an intrinsic genetic instability . Here , we investigated whether P11274 / P00519 expression could modulate nucleotide excision repair ( P55055 ) . We found that ectopic expression of Q92817 P11274 / P00519 in murine lymphoid BaF3 cell line inhibited P55055 activity in vitro , promoting hypersensitivity of these cells to ultraviolet ( UV ) treatment and facilitating a mutator phenotype . However , expression of Q92817 P11274 / P00519 in human and murine myeloid cell lines and primary bone marrow cells resulted in the increased P55055 activity and resistance to UV irradiation . The P00519 tyrosine kinase inhibitor STI571 reversed these effects , showing that Q92817 P11274 / P00519 tyrosine kinase activity is responsible for deregulation of P55055 . Hypoactivity of P55055 in Q92817 P11274 / P00519 -positive lymphoid cells was accompanied by the decreased interaction between proliferating cell nuclear antigen ( P12004 ) and xeroderma pigmentosum group B ( P19447 ) ; conversely , this interaction was enhanced in Q92817 P11274 / P00519 -positive myeloid cells . P78357 P11274 / P00519 did not affect P55055 in lymphoid and myeloid cells . In summary , our study suggests that Q92817 P11274 / P00519 reduced P55055 activity in lymphoid cells , leading to hypersensitivity to UV and mutagenesis . In contrast , Q92817 P11274 / P00519 expression in myeloid cells facilitated P55055 and induced resistance to UV . Chronic myeloid leukemic cells trigger poly(ADP-ribose) polymerase-dependent inactivation and cell death in lymphocytes . NK cells and T cells are commonly dysfunctional in CML , and their status may determine the course of disease . We aimed to define the molecular mechanisms of leukemia-induced immunosuppression with focus on the role of ROS and the P09874 pathway of cell death . Malignant granulocytes from patients with P11274 - P00519 -positive CML expressed the oxygen radical-producing enzyme NOX , produced large amounts of ROS , and triggered extensive cell death in NK cells . Inhibition of P09874 maintained NK cell viability in cocultures with suppressive leukemic cells . Under conditions of oxidative stress , P09874 inhibition upheld the capacity of NK cells to kill myeloid leukemic cells , in addition to restoring the proliferation and cytokine production of NK cells and cytotoxic T cells . Our findings are suggestive of a novel pathway of relevance to immunosuppression in CML . Deficiency of a phosphatidylinositol-anchored cell adhesion molecule influences haemopoietic progenitor binding to marrow stroma in chronic myeloid leukaemia . The interactions between haemopoietic progenitor cells and marrow stromal cells that are essential for the regulation of normal haemopoiesis are defective in chronic phase chronic myeloid leukaemia ( CML ) . The presence of primitive progenitor cells ( blast colony-forming cells , Bl- Q15814 ) in the blood of patients with CML is reflected by their reduced capacity to bind to marrow derived stromal layers in vitro . Whereas normal bone marrow Bl- Q15814 bind irreversibly to cultured stromal layers ( and none are found in normal blood ) , the Bl- Q15814 in CML bind transiently and then detach . The normal cell adhesion mechanism is partially sensitive to treatment with phosphatidylinositol-specific phospholipase C ( Pl- P98160 ) , indicating the participation of a phosphatidylinositol ( Pl ) -linked structure ; however , when CML cells were treated with Pl- P98160 it had no effect on progenitor binding . Two other Pl-linked structures , decay-accelerating factor ( P08174 ) and lymphocyte function associated antigen-3 ( LFA-3 ) were normally expressed on P28906 positive CML cells and normally susceptible to Pl- P98160 treatment . The treatment of normal cells with Pl- P98160 , to mimic the situation in CML , resulted in the indiscriminate and inefficient binding of Bl- Q15814 to stroma . Moreover , treatment of the normal cells with 5637 conditioned medium ( CM ) , which contains haemopoietic growth factors , also reduced the binding capacity of normal Bl- Q15814 ; 5637CM treatment did not alter the expression of P08174 . It is proposed that a Pl-linked cell adhesion molecule ( P62158 ) is deficient in CML as a consequence of the constitutive activation of P00519 kinase whilst , in normal cells , CAMs attached in this manner are responsible for efficient adhesion to stroma and are regulated by growth factors . 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 . 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 . 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 . Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . Comparison of the in vivo activity of different oxytocin antagonists in the pregnant baboon . OBJECTIVE : To ascertain the relative activity of five oxytocin antagonists ( OTAs ) in vivo in a tethered pregnant baboon model and compare these results to previously reported affinities in human and rat oxytocin receptor assays and median effective dose in rat uterotonic bioassays . METHODS : Pregnant tethered baboons between days 130 and 160 of pregnancy were given an oxytocin challenge test 1 minute after infusion of 1 mg of one of five randomly selected OTAs : ANTAG I , ANTAG II , ANTAG III , L366948 , and Atosiban . Once the uterine response to oxytocin returned to normal ( 1-8 days ) the O75051 was repeated with one of the remaining , untested OTAs during the 130-160 day period . Uterine activity , the time until the first significant response , and the dose of oxytocin needed to induce this response were all factored into one expression , the antagonist-response interval ( Q9Y4X5 ) . RESULTS : When expressed as ratio to ANTAG I the relative Q9Y4X5 for the OTAs were 0 , .5 , 1.0 , 2.4 and 59.2 for L366948 , Atosiban , ANTAG I , ANTAG II , and ANTAG III , respectively . ANTAG III and L366948 were significantly different from each other and the three other OTAs ( P < .05 ) . The log10 Q9Y4X5 for the 4 active OTAs when correlated with the log10 of the human and rat oxytocin receptor affinities and the rat uterotonic bioassay were all highly correlated ( r = .99 ; P < .05 ) . CONCLUSION : ANTAG III is a potent , long-acting OTA in vivo in the pregnant baboon and has the potential as a tocolytic in humans . Evaluation of antileukaemic effects of rapamycin in patients with imatinib-resistant chronic myeloid leukaemia . BACKGROUND : Recent data suggest that the mammalian target of rapamycin ( P42345 ) is involved in the regulation of growth of neoplastic cells in chronic myeloid leukaemia ( CML ) . PATIENTS AND METHODS : We treated six patients with imatinib-resistant CML in haematological relapse ( leukocytes > 20,000 microL(-1) ) with rapamycin at 2 mg per os daily for 14 consecutive days , with dose-adjustment allowed to reach a target rapamycin serum concentration of 10-20 pg mL(-1) . RESULTS : A major leukocyte response with decrease to less than 10,000 microL(-1) was obtained in two patients , and a minor transient response was seen in two other patients . In responding patients , we also observed a decrease in vascular endothelial growth factor ( P15692 ) mRNA levels in circulating leukaemic cells . Side effects during rapamycin treatment were mild in most patients . In one patient , pneumonia developed . DB00877 was also found to counteract growth of CML cells in vitro as determined by (3)H-thymidine incorporation . Moreover , rapamycin inhibited the growth of Ba/ P13726 cells exhibiting various imatinib-resistant mutants of P11274 / P00519 , including the T315I variant that exhibits resistance against most currently available P11274 / P00519 kinase inhibitors . CONCLUSIONS : DB00877 shows antileukaemic effects in imatinib-resistant CML in vitro and in vivo . Larger trials with rapamycin or rapamycin-derivatives in combination with other targeted drugs are warranted to further determine clinical efficacy in CML . Vasopressinergic regulation of the hypothalamic pituitary adrenal axis and stress adaptation . DB00067 ( VP ) stimulates pituitary DB01285 secretion through interaction with receptors of the V1b subtype ( P47901 , V3R ) , located in the plasma membrane of the pituitary corticotroph , mainly by potentiating the stimulatory effects of corticotropin releasing hormone ( P06850 ) . Chronic stress paradigms associated with corticotroph hyperresponsiveness lead to preferential expression of hypothalamic VP over P06850 and upregulation of pituitary P47901 , suggesting an important role for VP during adaptation of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis to stress . Vasopressinergic regulation of DB01285 secretion depends on the number of V1bRs as well as coupling of the receptor to phospholipase C ( P98160 ) in the pituitary . Regulation of P47901 gene transcription may involve a number of regulatory elements in the promoter region , of which a GAGA box was shown to be essential . Although P47901 gene transcription is necessary to maintain P47901 mRNA levels , the lack of correlation between VP binding and P47901 mRNA suggests that regulation of mRNA translation is a major regulatory step of the number of V1bRs . P47901 translation appears to be under tonic inhibition by upstream minicistrons and positive regulation through protein kinase C ( PKC ) activation of an internal ribosome entry site ( IRES ) in the 5' untranslated region ( 5'UTR ) of the mRNA . The data provide mechanisms by which regulation of hypothalamic VP and pituitary P47901 content contribute to controlling Q9Y251 axis activity during chronic stress . Effect of the hemoregulatory peptide (pEEDCK)2 (pyroGlu- DB00142 - DB00128 - DB00151 -Lys)2 and MIP-1alpha is reduced in bone marrow cultures from patients with chronic myeloid leukemia ( CML ) . The granulocyte-derived hemoregulatory peptide pyroGlu- DB00142 - DB00128 - DB00151 -Lys = pEEDCK is known to keep hematopoietic cells quiescent . When oxidized to its dimeric form (pEEDCK)2 , it activates growth of hematopoietic progenitors in association with stroma-derived cytokines . (pEEDCK)2 has a DB00151 - DB00151 motif which is also a typical feature of the macrophage inflammatory protein ( MIP-1alpha ) . The present study was designed to analyze differences between the response of normal and leukemic progenitor cells to (pEEDCK)2 or MIP-1alpha . When long-term bone marrow cultures ( LTBMCs ) were incubated with (pEEDCK)2 or MIP-1alpha and/or cytokines , the stimulatory effect on colony-forming units-granulocyte/erythroid/macrophage/megakaryocyte of LTBMC from chronic myeloid leukemia ( CML ) patients was less than 50 % compared to LTBMC from healthy humans . No difference in oncogene expression could be observed in LTBMC from CML patients regarding reduction of Philadelphia chromosome-associated transcription of the P11274 - P00519 gene . With respect to the expression of growth and differentiation-associated genes ( Galpha16 , P09917 , phospholipaseA2 , c-kit , and P28906 ) , which were analyzed from LTBMC by semiquantitative reverse transcriptase-polymerase chain reaction , the same transcription rate was observed in CML patients and healthy donors . However , two isoforms of a key enzyme of oxidative metabolism , carnitine palmitoyltransferase ( P50416 and Q92523 ) , showed 50-fold higher expression rates in LTBMC cells of healthy donors compared to CML patients . It is known that a decrease in oxidative metabolism is associated with an increase in redox equivalents in malignancy . This might result in a reduction of disulphide bonds in (pEEDCK)2 or MIP-1alpha , thus inducing a downregulation of these factors in bone marrow from CML patients .

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interacts_with DB00559?

multiple_choice

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 . Cytochrome P450 17 ( P05093 ) is involved in endometrial cancinogenesis through apoptosis and invasion pathways . Cytochrome P450 17 ( P05093 ) encodes cytochrome P450c17α , an enzyme with 17α-hydroxylase and 17 , 20-lyase activities involved in estradiol biosynthesis . Here we examine the role of P05093 gene in endometrial carcinogenesis . Immunohistochemistry staining of endometrial carcinoma and corresponding uninvolved tissues showed that P05093 is upregulated in endometrial cancers ( 15 of 24 , 62.5 % ) . To understand the functional significance of this upregulation , we silenced P05093 gene by introduction of siRNA into endometrial cancer cell line KLE followed by functional studies . Further , to understand the molecular basis of the role of P05093 , we profiled the expression of key pathway-specific genes and identified several components of the apoptosis and invasion pathways that are potentially regulated by P05093 . Silencing of P05093 caused decreased cell proliferation and induced apoptosis . Significantly , P05093 depletion leads to downregulation of anti-apoptotic genes B cell lymphoma 2 ( Bcl-2 ) and telomerase reverse transcriptase ( O14746 ) , indicating induced apoptosis . Also , attenuation of P05093 decreased the cellular invasion ability and regulated expression of several invasion pathway components such as melanoma cell adhesion molecule ( P43121 ) , matrix metallopeptidase 2 and 9 ( P08253 and P14780 ) , and tissue inhibitor of metalloproteinase 3 ( P35625 ) . In conclusion , this is the first report documenting that upregulation of P05093 in endometrial cancers play a crucial role in endometrial carcinogenesis by targeting multiple components of apoptosis and invasion pathways . Further studies are required to understand the detailed mechanisms underlying P05093 -mediated regulation of these components . Effect of abiraterone acetate plus prednisone on the QT interval in patients with metastatic castration-resistant prostate cancer . PURPOSE : DB05812 is the active metabolite of the pro-drug abiraterone acetate ( AA ) and a selective inhibitor of P05093 , a key enzyme in testosterone synthesis , and improves overall survival in postdocetaxel metastatic castration-resistant prostate cancer ( mCRPC ) . This open-label , single-arm phase 1b study was conducted to assess the effect of AA and abiraterone on the QT interval . METHODS : The study was conducted in 33 patients with mCRPC . Patients received AA 1,000 mg orally once daily + prednisone 5 mg orally twice daily . Electrocardiograms ( ECGs ) were collected in triplicate using 12-lead Holter monitoring . Baseline ECGs were obtained on Cycle 1 Day-1 . Serial ECG recordings and time-matched pharmacokinetic ( PK ) blood samples were collected over 24 h on Cycle 1 Day 1 and Cycle 2 Day 1 . Serial PK blood samples were also collected over 24 h on Cycle 1 Day 8 . RESULTS : After AA administration , the upper bound of the 2-sided 90 % confidence interval ( CI ) for the mean baseline-adjusted QTcF change was < 10 ms ; no patients discontinued due to QTc prolongation or adverse events . No apparent relationship between change in QTcF and abiraterone plasma concentrations was observed [ estimated slope ( 90 % CI ) : 0.0031 ( -0.0040 , 0.0102 ) ] . CONCLUSIONS : There is no significant effect of AA plus prednisone on the QT/QTc interval in patients with mCRPC . 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 . 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 . Abnormal NF-kappaB signaling pathway with enhanced susceptibility to apoptosis in immortalized keratinocytes . The transcriptional activation and proper regulation of NF-kappaB is known to be important to the apoptotic resistant phenotype of epidermal-derived keratinocytes . By comparing and contrasting the responses of normal foreskin-derived keratinocytes versus an immortalized skin-derived keratinocyte cell line ( i.e. HaCaT cells ) , several molecular defects involving NF-kappaB signaling pathway were delineated in the immortalized keratinocytes . While exposure to P01579 plus TPA produces growth arrest in both normal and immortalized keratinocytes , with rapid phosphorylation of MEKKI and recruitment of distinctive protein kinase C isoforms into the signalosome complex , subsequent molecular events necessary for NF-kappaB activation were abnormal in HaCaT cells . This disrupted NF-kappaB activation in HaCaT cells was accompanied by enhanced susceptibility to UV-light induced apoptosis , which was associated with elevated levels of Q01094 and decreased Q13077 /TRAF2 levels . Additional defects in HaCaT cells included markedly diminished levels of IKKbeta ( and lack of induction of kinase activity ) in response to inflammatory stimuli , a failure of P38936 ( P38936 /CIP1) to associate with P24941 , and a decreased association between p65 and p300 . These studies suggest caution in using HaCaT cells as a substitute for normal keratinocytes to study apoptosis in the skin . Thus , it appears that while the immortalized cells can escape cell cycle checkpoints by elevated levels of Q01094 , an adverse biological consequence of such dysregulated cell cycle control is the inability to activate the anti-apoptotic NF-kappaB signaling pathway . Therefore , exploiting this apoptosis vulnerability in pre-malignant , or immortalized cells , prior to acquiring a death-defying phenotype characteristic of more advanced malignant cell types , provides the basis for an early interventional therapeutic strategy for cutaneous oncologists . Multi-chaperone complexes regulate the folding of interferon-gamma in the endoplasmic reticulum . The quality control mechanisms directing the folding of cytokines in the endoplasmic reticulum ( ER ) are poorly understood . We have investigated ER chaperone usage by the cytokine interferon-gamma ( P01579 ) . DB00171 -depletion or inhibition of N-glycosylation was found to cause P01579 to accumulate into detergent-insoluble aggregates in the ER . Six chaperones , P14625 , P11021 , P13667 , P07237 , Q9NZU7 / Q15084 and CRT were found to associate with P01579 during its steady state folding . Interaction of the five first chaperones with P01579 was regulated co-ordinately by DB00171 . These chaperones were recently reported to be part of a multi-chaperone complex involved in the folding of complex , multi-subunit proteins . Our data suggest that also proteins with a relatively simple quaternary structure such as cytokines may fold in association with this complex . In addition , we identified calreticulin as the major chaperone interacting with P01579 , and the related class II cytokine interleukin-10 , during heat-shock in vivo . P01579 was maintained in a folding-competent form by calreticulin during heat-shock and released during subsequent recovery at 37 degrees C . This interaction was observed in both recombinant ( CHO- P03951 ) and natural producer cells ( Jurkat , NK-92MI ) of P01579 . Since cytokines such as P01579 and P22301 are frequently produced in the course of inflammatory conditions associated with fever , the thermo-protective effect of calreticulin may constitute a previously unrecognized component of the cellular cytokine production machinery , of likely relevance in sustaining cytokine folding and secretion in pathophysiological conditions . 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 . Task-dependent interactions between dopamine D2 receptor polymorphisms and DB01235 in patients with Parkinson 's disease . Variants in genes regulating dopamine transmission affect performance on tasks including working memory and executive function as well as temporal processing and sequence learning . In the current study , we determined whether a dopamine D2 receptor DNA sequence polymorphism interacts with DB01235 during motor tasks in patients with Parkinson 's disease ( PD ) . Forty-five PD patients were genotyped for the P14416 polymorphism ( rs 1076560 , G > T ) . Patients performed an explicit motor sequence learning task and the grooved pegboard test in both ON and OFF DB01235 states . For motor sequence learning , P14416 genotype mediated DB01235 effects such that DB01235 associated improvements were only observed in the minor T allele carriers ( associated with lower D2 receptor availability , t10=-2.71 , p=0.022 ) , whereas G homozygotes showed no performance change with DB01235 . For the grooved pegboard test , performance improved with DB01235 independent of patients ' P14416 genotype . Collectively these results demonstrate that common P14416 allelic differences found in the human population may explain how dopamine differentially contributes to performance across tasks and individuals . 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 . P35354 inhibitors and metabolism of essential fatty acids . Selective P35354 inhibitors increase the risk of myocardial infarction and stroke that is attributed to their ability to inhibit prostacyclin ( DB01240 ) , lipoxins , resolvins , and endothelial nitric oxide ( eNO ) but not platelet P23219 derived thromboxane A2 ( TXA2 ) . In contrast , aspirin blocks both P23219 and P35354 enzymes that , in turn , increases intracellular concentrations of dihomo-gamma-linolenic acid ( DB00154 ) , arachidonic acid ( AA ) , eicosapentaenoic acid ( EPA ) and docosahexaenoic acid ( DB01708 ) and reduced formation of eicosanoids . On the other hand , such an increase is much less with specific P35354 inhibitors since they do not block the formation of eicosanoids through P23219 pathway . DB00154 , AA and EPA form precursors to PGE1 , DB01240 , and PGI3 respectively , which are potent vasodilators and platelet anti-aggregators , and thus aid in the prevention of thrombus formation . EPA has anti-arrhythmic action , and EPA , DB01708 ( docosahexaenoic acid ) , DB00154 , and PGE1 have anti-inflammatory actions as well . EPA , DB01708 , and AA augment eNO formation that has anti-atherosclerotic action . Hence , combining EFAs with P35354 inhibitors will prevent thrombotic cardiovascular events . Human kidney : endothelin isoforms detected by HPLC with radioimmunoassay and receptor subtypes detected using ligands BQ123 and BQ3020 . Animal kidneys are exquisitely sensitive to the effects of endothelin ( ET ) , but little is known of its binding characteristics , isoform prevalence , or receptor subtype distribution in human kidney . We investigated these parameters using high-performance liquid chromatography , radioimmunoassay ( RIA ) , and the recently synthesized P25101 and ETB receptor-selective peptide ligands BQ123 ( cyclo[D- DB00128 -L-Pro-D- DB00161 -L- DB00149 -D- DB00150 -] ) and BQ3020 ( Ala11,15-Ac-ET-1[6-21] ) . Fresh-frozen normal segments of kidneys excised for carcinoma were solid-phase-extracted using Amprep P06681 columns , and parallel eluates were oxidized . All were subjected to RIA for ET and pro-ET-1 , in triplicate . ET isoforms were characterized by RP-HPLC and subsequent RIA of eluates . Total amounts of immunoreactive ET were 6.9 +/- 3.8 and 4.5 +/- 1.6 pmol/g wet weight in medulla and cortex , respectively . Reverse-phase high performance liquid chromatography showed peaks of immunoreactivity with retention times identical to synthetic ET-1 and metsulphoxide ET-1 . P20800 , P14138 , and pro-ET-1 were not detected . Saturation assays using 0.01-8.0 nM 125I-ET-1 or 125I-BQ3020 , gave Kd values ( mean +/- SEM ) of 0.17 +/- 0.04 and 0.36 +/- 0.06 nM , respectively , with Bmax values of 57.7 +/- 15.4 and 30.0 +/- 5.0 fmol/mg protein , respectively . Hill coefficients were 0.86 +/- 0.03 and 0.77 +/- 0.04 , but a two-site fit was not preferred . Receptor autoradiography has detected both subtypes , mainly present in medulla , with ETB predominating. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . 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 . P00505 from rat heart is phosphorylated on Tyr19 in response to insulin stimulation . The cytosolic fatty acid-binding protein from rat heart ( P05413 , M(r) 15,000 ) as well as FABP from mouse adipocytes ( P15090 , 62 % homologous with P05413 ) contain a recognition sequence for protein tyrosine kinases , DB00174 - DB00120 - DB00128 - DB00128 -Tyr19 . P15090 has been shown by others to be partly phosphorylated on Tyr19 , thus encouraging experiments designed to search for phosphotyrosine in P05413 . For this purpose isolated cardiac myocytes were incubated with [32P]orthophosphate and analyzed by two-dimensional gel electrophoresis . A 15 kDa phosphoprotein present in the cytosolic protein fraction was specifically precipitated by a polyclonal antibody against rat heart FABP . Characterization of the phosphorylated FABP was facilitated by the development of an immunoaffinity purification procedure capable of isolating more than 200 micrograms FABP from four rat hearts in one step . Phosphoamino acid analysis and radiosequencing of the major tryptic phosphopeptide from immunopurified FABP revealed Tyr19 as the phosphorylated amino acid . Stimulation of cardiac myocytes with insulin in the presence of tyrosine phosphatase inhibitors led to a several-fold increase in the amount of phosphorylated FABP compared with a nearly undetectable level found without insulin stimulation , indicating that FABP may be a substrate for the insulin receptor tyrosine kinase . Phosphorylated FABP constitutes only a minor fraction compared to the large pool of FABP in the cardiac myocyte , thus obscuring the significance of this modification . However , as the phosphorylated Tyr19 residue is positioned within a helix-turn-helix-related domain of FABP , this modification may modulate a hitherto unknown DNA binding activity of FABP . A hypothesis is discussed in which phosphorylated FABP serves as a signalling molecule in the insulin signal transduction cascade . Arrestins as signaling modulators : the plot thickens : focus on " Arrestins 2 and 3 differentially regulate P25101 and P41231 receptor-mediated cell signaling and migration in arterial smooth muscle " . [ Localization of the NotI clones from human chromosome 3 on quail microchromosomes ] . For the purpose of comparative mapping of quail ( Coturnix c. japonica ) and human ( Homo sapiens ) genomes , DNA fragments from human chromosome 3 ( HSA3p14-21 and HSA3q13-23 ) were localized on quail mitotic chromosomes . Using the method of double-color fluorescence DNA-DNA in situ hybridization , these fragments were mapped to two different microchromosomes . Earlier , similar studies were performed using chicken mitotic chromosomes . There it was demonstrated that the clones of interest were distributed among three microchromosomes ( GGA12 , GGA14 , and GGA15 ) . Thus , interspecific difference in the location of human chromosome 3 DNA fragments in the genomes of closely related avian species was discovered . A new confirmation of the hypothesis on the preferable localization of the gene-rich human chromosome regions on avian microchromosomes was obtained . At the same time , a suggestion on the localization of some orthologous genes in the genome of the organism under study was made : P18085 , Q14524 , Q9BWX1 , Q9BV23 , Q9NYG2 , Q16644 , ADSYNA ( homolog of chicken chromosome 12 ) , P14416 , PP2C- P25101 , P51149 , P32238 , and PKD1 ( homolog of chicken chromosome 15 ) . However , localization of the corresponding quail genes needs to be confirmed , as far as the sequences used were only the orthologs of the corresponding chicken genes . 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 . 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 . 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 . Adipocyte fatty acid-binding protein as a determinant of insulin sensitivity in morbid-obese women . The aim of the study was to evaluate human plasma circulating levels of adipocyte fatty acid-binding protein ( P15090 ) and its relationship with proinflammatory adipocytokines and insulin resistance in a severely obese cohort , before and 1 year after a surgical gastric bypass . Plasmatic levels of P15090 were measured in 77 morbid-obese women before and 1 year after bariatric surgery . Anthropometrical parameters and body composition by bioelectrical impedance analysis were determined . Circulating levels of soluble tumor necrosis factor receptor 2 ( sTNFR2 ) , Interleukin 18 ( Q14116 ) , adiponectin , and high-sensitive P02741 ( hsCRP ) were also analyzed . P01308 resistance by homeostasis model assessment of insulin resistance ( HOMA-IR ) index was calculated . After massive weight loss , P15090 plasmatic levels decreased significantly [ 7.6 ( 8.9 ) vs. 4.3 ( 5.1 ) ; P < 0,001 ] but no association with circulating adipokines or proinflammatory cytokines , both at the beginning and at the end of follow-up , was observed . A decrease in sTNFR2 , Q14116 , hsCRP , and an increase in adiponectin levels ( P < 0.001 in all cases ) were observed after the gastric bypass . HOMA-IR index improved 1 year after surgery and after multiple regression analysis remained associated with P15090 after controlling for confounding variables ( beta=0.322 , P=0.014 ; R2 for the model 0.281 ) . In morbid-obese women , plasma P15090 concentrations were dramatically reduced after gastric bypass surgery . After weight loss this protein contributed to HOMA-IR index independently of proinflammatory/antinflammatory cytokine profile . Further studies are warranted to elucidate the role of P15090 in the pathogenesis of insulin resistance in morbid obesity . Altered endothelin receptor expression and affinity in spontaneously hypertensive rat cerebral and coronary arteries . BACKGROUND : Hypertension is associated with arterial hyperreactivity , and endothelin ( ET ) receptors are involved in vascular pathogenesis . The present study was performed to examine the hypothesis that ET receptors were altered in cerebral and coronary arteries of spontaneously hypertensive rats ( SHR ) . METHODOLOGY/PRINCIPAL FINDINGS : Cerebral and coronary arteries were removed from SHR . Vascular contraction was recorded using a sensitive myograph system . Real-time PCR and Western blotting were used to quantify mRNA and protein expression of receptors and essential MAPK pathway molecules . The results demonstrated that both P25101 and ETB receptor-mediated contractile responses in SHR cerebral arteries were shifted to the left in a nonparallel manner with increased maximum contraction compared with Wistar-Kyoto ( WKY ) rats . In SHR coronary arteries , the P25101 receptor-mediated contraction curve was shifted to the left in parallel with an increased pEC50 compared with the arteries in WKY rats . There was no significant increase in ETB receptor-mediated contraction in SHR coronary arteries . P25101 receptor mRNA and protein expression was increased in SHR cerebral arteries compared with the arteries in WKY rats . However , P25101 receptor mRNA and protein levels in coronary arteries and ETB receptor protein levels in cerebral and coronary arteries remained unchanged in SHR compared with WKY rats . Meanwhile , phosphorylated P27361 /2 protein was significantly increased in SHR brain and heart vessels . CONCLUSIONS/SIGNIFICANCE : In SHR cerebral arteries , P25101 receptor expression was upregulated . P25101 receptor affinity was increased in coronary arteries , and ETB receptor affinity was increased in cerebral arteries . The P27361 /2 activation may be involved in the receptor alterations .

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interacts_with DB06616?

multiple_choice

Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . [ Therapeutic use of anti- P01375 agents in spondyloarthropathies ] . CLASSICAL DATA : Spondyloarthropathies regroup several rheumatological entities ( ankylosing spondylitis , reactive arthritis , psoriatic rheumatism , entero-colopathic disease rheumatism , undifferentiated spondyloarthropathies ) with validated diagnosis criteria . Drug therapy is based upon NSAIDs ( non-steroidal antiinflammatories ) . Refractory forms may lead to severe functional impairment , raising the need of more effective treatments . IN FAVOUR OF ANTI- P01375 -ALPHA AGENTS : Several arguments ( P01375 serum levels , elevated levels of mRNA , P01375 messengers , in sacro iliac biopsies , efficacy of anti P01375 agents in Crohn 's disease ) justify the use of anti- P01375 agents in the treatment of spondyloarthropathies . Two biologic agents have been assessed in these circumstances : a monoclonal antibody ( DB00065 ) and a soluble form of the P01375 receptor ( DB00005 ) . EFFICACY AND SAFETY : Results of open and controlled studies , although on small series , demonstrated the significant efficacy of anti- P01375 agents on the various clinical , biological , functional and quality of life parameters , and confirmed by imaging ( Q9BWK5 ) . Tolerance is fair , but two cases of diffuse tuberculosis have been reported with DB00065 . THERAPEUTIC PROGRESS : Even if additional studies are required to answer some questions ( long term efficacy and safety , treatment modalities ) , anti P01375 agents appear as a therapeutic progress in refractory spondyloarthropathies , for which few validated options have existed up till now . 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 . [ Expression of P11274 / P00519 fusion gene in circulating endothelial cells from chronic myelogenous leukemia patients and its clinical significance ] . Several studies have shown that the tumor endothelial cells are different from the normal tissue endothelial cells . These tumor endothelial cells may contribute to tumor neo-vasculogenesis . This study was purposed to analyze the biologic features and determine the expression level of CD133 and P11274 / P00519 fusion gene in circulating endothelial cells ( CEC ) isolated from peripheral blood of CML patients , as well as to investigate the role of CEC in disease progression . Mononuclear cells were isolated from peripheral blood by density gradient centrifugation ; CEC were sorted by P29966 and harvested in the endothelial growth medium . The morphologic features of CEC were observed by microscopy , the cell growth rate was calculated by cell counting , and the cells were identified by immunofluorescence staining for the expression of CD31, P28906 , P04275 and CD133 . The expression of P11274 / P00519 fusion gene was examined by Q5TCZ1 in 12 CML patients . The results indicated that the isolated CEC displayed the typical cobble-stone morphology . These cells could be identified by the positive immunofluorescence staining for CD31 , P28906 and P04275 , and showed more increased proliferative potential as compared to that of healthy donors . It was found that the positive rate of CD133 was 31.29 % in CML patients , which was significantly different from that of healthy donors ( P < 0.05 ) . In 12 CML patients , CEC carried the same chromosome aberration as the leukemia cells ( 10.77 % ) . Higher expression level of CD133 and P11274 / P00519 fusion gene positively correlated with progression of disease . It is concluded that the CEC may participate in invasion and angiogenesis in patients with CML and possibly correlate to the spreading and progression of the disease . 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 . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Neuroprotective profile of novel P12931 kinase inhibitors in rodent models of cerebral ischemia . Src kinase signaling has been implicated in multiple mechanisms of ischemic injury , including vascular endothelial growth factor ( P15692 ) -mediated vascular permeability that leads to vasogenic edema , a major clinical complication in stroke and brain trauma . Here we report the effects of two novel Src kinase inhibitors , 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile ( DB06616 ) and 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[4-(4-methypiperazin-1-yl)but-1-ynyl]-3-quinolinecarbonitrile ( SKS-927 ) , on ischemia-induced brain infarction and short- and long-term neurological deficits . Two well established transient [ transient middle cerebral artery occlusion ( tMCAO ) ] and permanent [ permanent middle cerebral artery occlusion ( pMCAO ) ] focal ischemia models in the rat were used with drug treatments initiated up to 6 h after onset of stroke to mimic the clinical scenario . Brain penetration of Src inhibitors , their effect on blood-brain barrier integrity and P15692 signaling in human endothelial cells were also evaluated . Our results demonstrate that both agents potently block P15692 -mediated signaling in human endothelial cells , penetrate rat brain upon systemic administration , and inhibit postischemic Src activation and vascular leakage . Treatment with DB06616 or SKS-927 ( at the doses of 3-30 mg/kg i.v. ) resulted in a dose-dependent reduction in infarct volume and robust protection from neurological impairments even when the therapy was initiated up to 4- to 6-h after tMCAO . Src blockade after pMCAO resulted in accelerated improvement in recovery from motor , sensory , and reflex deficits during a long-term ( 3 weeks ) testing period poststroke . These data demonstrate that the novel Src kinase inhibitors provide effective treatment against ischemic conditions within a clinically relevant therapeutic window and may constitute a viable therapy for acute stroke . Antiidiotype induction therapy : evidence for the induction of immune response through the idiotype network in patients with ovarian cancer after administration of anti- Q8WXI7 murine monoclonal antibody DB04964 . The immune status of ovarian cancer patients receiving anti- Q8WXI7 murine monoclonal antibody DB04964 was evaluated by measuring antiidiotypic antibodies ( Ab2 ) , antiantiidiotypic antibodies ( Ab3 ) , antiisotypic human antimouse antibodies ( HAMA ) , interferon-gamma , and Q8WXI7 levels in the serum . A specific assay was developed for the determination of Ab2 antibodies using chimeric MAb DB04964 . Of the 50 patients studied , 26 had elevated levels of Ab2 . Eleven of these 26 patients also had high titer of antiantiidiotypic ( Ab3 ) antibodies . Eight of the 22 patients analyzed had increased interferon-gamma levels . A tentative correlation was found between survival of these patients ' antiidiotype induction . Effect of atorvastatin on endothelial function and inflammation in long-duration type 1 diabetic patients without coronary heart disease and arterial hypertension . AIM : We evaluated the ability of atorvastatin , an P04035 inhibitor , to affect endothelial function and inflammation in long-duration ( > 10 years ) type 1 diabetes mellitus ( T1DM ) patients without coronary heart disease ( Q8NE62 ) and arterial hypertension ( AH ) . METHODS AND RESULTS : We randomized 204 Caucasians with long-duration T1DM into either the atorvastatin 40 mg/day plus hypolipaemic diet group ( n = 154 ) or the placebo plus hypolipaemic diet group ( n = 50 ) for 6 months . Endothelium-dependent flow-mediated ( FMD ) and endothelium-independent flow-mediated vasodilatation , serum levels of plasminogen activator inhibitor-1 ( P05121 ) , P04275 ( P04275 ) and high sensitivity P02741 ( hs-CRP ) were estimated before and after treatment . After 6 months of therapy , FMD was increased by 44 % in the atorvastatin plus diet group compared with the placebo plus diet group . Treatment with atorvastatin led to a significant reduction in levels of P05121 and hs-CRP ; however , the elevation of P04275 level was observed . In the placebo plus diet group , we observed a significant reduction in levels of hs-CRP but not of P04275 and P05121 . CONCLUSIONS : DB01076 improves endothelial function and reduces some proinflammatory and prothrombotic markers of atherosclerosis in T1DM patients without Q8NE62 and AH . The surprising effect of atorvastatin on serum P04275 levels in T1DM requires further study . 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 . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . P35367 activation stimulates mitogenesis in human astrocytoma U373 MG cells . In human astrocytoma U373 MG cells that express histamine H1 receptors ( 180 +/- 6 fmol/mg protein ) but not H2 or H3 receptors , histamine stimulated mitogenesis as assessed by [ 3H ] -thymidine incorporation ( 173 +/- 2 % of basal ; EC50 , 2.5 +/- 0.4 microM ) . The effect of 100 microM histamine was fully blocked by the selective H1 antagonist mepyramine ( 1 microM ) and was markedly reduced ( 93 +/- 4 % inhibition ) by the phospholipase C inhibitor U73122 ( 10 microM ) . The activator of protein kinase C ( PKC ) phorbol 12-tetradecanoyl-13-acetate ( TPA , 100nM ) stimulated [ 3H ] -thymidine incorporation ( 270 +/- 8 % of basal ) , and this response was not additive with that to 100 microM histamine . The incorporation of [ 3H ] -thymidine induced by 100 microM histamine was partially reduced by the PKC inhibitor Ro 31-8220 ( 57 +/- 7 % inhibition at 300 nM ) and by the compound PD 098,059 ( 30 microM , 62 +/- 14 % inhibition ) , an inhibitor of the mitogen-activated kinase ( MAPK ) kinases Q02750 / P36507 . These results show that histamine H1 receptor activation stimulates the proliferation of human astrocytoma U373 MG cells . The action of histamine appears to be partially mediated by PKC stimulation and MAPK activation . 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 . Phorbol 12-Myristate 13-Acetate Induces Q8WXI7 Expression via PKCδ and p38 in Human Airway Epithelial Cells . OBJECTIVES : Phorbol 12-myristate 13-acetate ( PMA ) is widely used as a protein kinase C ( PKC ) activator , PKC is involved in the secretion of mucins . Q8WXI7 , one of the membrane-bound mucins , is produced in human airway epithelial cells . However , the effect and signaling pathway of PMA on Q8WXI7 expression in human airway epithelial cells has not been reported . Therefore , the effect and brief signaling pathway of PMA on Q8WXI7 expression were investigated in human airway epithelial cells in this study . METHODS : In the mucin-producing human NCI-H292 airway epithelial cells and the primary cultures of normal nasal epithelial cells , the effect and signaling pathway of PMA on Q8WXI7 expression were investigated using reverse transcriptase-polymerase chain reaction ( RT-PCR ) , real-time PCR , enzyme immunoassay , and immunoblot analysis with several specific inhibitors and small interfering RNA ( siRNA ) for p38 mitogen-activated protein kinase ( MAPK ) . RESULTS : PMA increased Q8WXI7 expression , and activated phosphorylation of p38 MAPK . However , it did not activate phosphorylation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . SB203580 ( p38 MAPK inhibitor ) inhibited PMA-induced Q8WXI7 expression , while U0126 ( P27361 /2 inhibitor ) did not . In addition , the knockdown of p38 MAPK by p38 MAPK siRNA significantly blocked PMA-induced Q8WXI7 mRNA expression . Rottlerin ( PKCδ inhibitor ) inhibited PMA-induced Q8WXI7 expression , and also inhibited the phosphorylation of activated p38 MAPK by PMA . CONCLUSION : These results show for the first time that PMA-induced Q8WXI7 expression is regulated by activation of the PKCδ and p38 MAPK signaling pathway in human airway epithelial cells . The arylstibonic acid compound NSC13746 disrupts B- Q8N5A5 binding to DNA in living cells . The inhibition of DNA binding of basic leucine zipper ( B- Q8N5A5 ) transcription factors is a clinically relevant molecular target . Our laboratory has previously reported two methods of inhibiting B- Q8N5A5 DNA binding in solution : 1 ) an arylstibonic acid compound that binds to the basic region , stabilizes the B- Q8N5A5 dimer , and prevents B- Q8N5A5 DNA binding and 2 ) dominant negative proteins , termed A-ZIPs , that heterodimerize with B- Q8N5A5 domains in a leucine zipper-dependent manner . To determine if these two agents also inhibit DNA binding in live cells , GFP-tagged B- Q8N5A5 domains and mCherry-tagged A- Q8N5A5 domains were transfected into NIH3T3 cells to assess protein localization and Fluorescence Recovery After nuclear Photobleaching ( P42345 ) . P42345 , showed that all six GFP-B- Q8N5A5 domains examined recovered faster in the nucleus in the presence of drug that we interpret represents an inhibition of DNA binding . Faster recovery in the presence of the A- Q8N5A5 was leucine zipper dependent . The arylstibonic also induced a cytoplasmic localization of all B- Q8N5A5 domains while the A-ZIPs induced a leucine zipper-dependent cytoplasmic localization . Thus , the change in cellular localization of B- Q8N5A5 domains could be used as a high-throughput assay for inhibitors of B- Q8N5A5 DNA binding . Additionally , the arylstibonic acid compound was cytostatic in clear cell sarcoma cells , which express a chimera between the B- Q8N5A5 domain of P39905 -1 and N-terminal activation domain of Q01844 but not in K562 cells that express a non-B- Q8N5A5 containing chimeric protein P11274 - P00519 . These studies suggest that arylstibonic acid compounds or other small molecules capable of inhibiting B- Q8N5A5 DNA binding could be valuable anticancer agents . 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 . Acetylbritannilactone suppresses lipopolysaccharide-induced vascular smooth muscle cell inflammatory response . To investigate the mechanism of action by which a new anti-inflammatory active compound , 1-O-acetylbritannilactone ( P00519 ) isolated from Inula britannica-F. , inhibits inflammatory responses in vascular smooth muscle cells ( VSMCs ) . Enzyme immunoassay was used to measure the levels of prostandin E(2) ( PGE(2) ) production . Immunocytochemistry staining and Western blot analysis were performed to detect the nuclear translocation of nuclear factor-kappaB ( NF-kappaB ) p65 and the expression of IkappaB-alpha , pIkappaB-alpha and cyclooxygenase-2 ( P35354 ) . Electrophoretic mobility shift assays ( EMSA ) were used to detect DNA-binding activity of NF-kappaB in VSMCs . P00519 ( 5 , 10 , 20 micrommol/l ) had several concentration-dependent effects , including inhibition of lipopolysaccharide ( LPS ) -induced PGE(2) production and P35354 expression , and blockade of NF-kappaB activation and translocation . These effects were owing to reductions in IkappaB-alpha phosphorylation and degradation induced by LPS . In addition , P00519 directly inhibited the binding of active NF-kappaB to specific DNA cis-element . These results indicate that P00519 is a potent inhibitor of LPS-stimulated VSMC inflammatory responses through blockade of NF-kappaB activity and inhibition of inflammatory gene P35354 expression . Gene transfer of GM- P04141 , P33681 and CD154 cDNA enhances survival in a murine model of acute leukemia with persistence of a minimal residual disease . Gene transfer of various cytokines and co-stimulatory molecules has been reported to induce a potent antileukemic immunity in murine models , however , the relative efficiency and possible synergistic effects between candidate genes have not been extensively investigated . We analyzed in a murine model of P11274 / P00519 acute leukemia whether gene transfer of CD154 , P33681 or GM- P04141 as a single agent or combination of CD154 + GM- P04141 , P33681 + CD154 and GM- P04141 + P33681 in leukemic cells could enhance survival . We observed that CD154 gene transfer induced a marked inhibition of leukemogenicity , and also that CD154 and combination of GM- P04141 and P33681 gene transfer protected mice against subsequent challenge with leukemic cells and had a therapeutic effect for a pre-established leukemia disease . We also found minimal residual leukemic disease by RT-PCR for 6 to 12 months in 0 to 25 % of animals injected with transduced leukemic cells and surviving the challenge without evidence of disease , except in the control empty plasmid group where very few mice survived the challenge but all of those were positive by RT-PCR . These findings suggest that leukemic cell vaccination by gene transfer can induce a tumor dormancy phenomenon compatible with long-term survival .

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interacts_with DB08827?

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 . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . DB08827 . Aegerion Pharmaceuticals is developing lomitapide , a small-molecule , microsomal triglyceride transfer protein ( P55157 ) inhibitor , for the treatment of both familial and primary hypercholesterolemia . Oral , once-daily lomitapide will be targeted at patients resistant to P04035 inhibitors ( statins ) either due to abnormalities in liver function or to discontinuation because of muscle pain . An oral formulation of lomitapide is in phase III development for homozygous familial hypercholesterolemia ( hyperlipoproteinemia type IIa ) in the US , Canada , Italy , and South Africa . This review discusses the key development milestones and therapeutic trials of this drug . Pharmacological modulation of myocardial tumor necrosis factor alpha production by phosphodiesterase inhibitors . Phosphodiesterase ( PDE ) inhibitors are used as therapeutic agents for management of congestive heart failure . PDE inhibitors are potent inotropic and vasodilator drugs , which have also been shown to inhibit tumor necrosis factor alpha ( P01375 ) production . P01375 is a pleiotropic cytokine that has the ability to produce cardiac depressant and other cardiovascular effects in many disease conditions . P01375 levels are elevated in patients with chronic congestive heart failure , and it is possible that P01375 may play a role in this condition . The effects of PDE inhibitors on P01375 secretion from rat heart were evaluated in this study . Rat left ventricle was minced and incubated for 4 hr with various PDE inhibitors , and the amount of P01375 secretion was evaluated by cytotoxicity assay . Ro-20 , 1724 , etazolate , amrinone , milrinone and pentoxifylline inhibited unstimulated P01375 production , with IC50 values of 1.87 , 2.07 , 13.9 , 153 and 201 microM , respectively . Lipopolysaccharide-induced P01375 secretion from rat left ventricle was also evaluated in this study . DB01427 , milrinone and pentoxifylline inhibited lipopolysaccharide-induced P01375 secretion , with IC50 values of 14.8 , 81.6 and 748 microM , respectively , whereas Ro-2D , 1724 and etazolate had no effect on lipopolysaccharide-induced P01375 secretion . These results demonstrated that P01375 was secreted from rat left ventricle after 4 hr and different pharmacological manipulations were able to inhibit the secretion of P01375 from left ventricle . These initial pharmacological results may provide an important tool for further investigation into the beneficial effects of PDE inhibitors in congestive heart failure or other conditions where P01375 levels are elevated . 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 ) Learnings from over 25 years of PNH experience : the era of targeted complement inhibition . Paroxysmal nocturnal haemoglobinuria ( PNH ) is a progressive and life-threatening disease that causes thrombosis , end organ damage and impaired quality of life . Chronic uncontrolled complement activation leads to chronic haemolysis , causing progressive morbidities and early mortality . Hence , early diagnosis is essential for improved patient management and prognosis . DB01257 ( SOLIRIS® ) specifically inhibits chronic , uncontrolled complement activation and is the first-in-class , humanised , monoclonal antibody targeting P01031 within the terminal complement pathway . DB01257 is the first and only approved treatment for PNH in adults and children . Computed tomography correlates with improvement with ivacaftor in cystic fibrosis patients with G551D mutation . BACKGROUND : DB08820 corrects the cystic fibrosis transmembrane conductance regulator ( P13569 ) gating defect associated with G551D mutation and is quickly becoming an important treatment in patients with cystic fibrosis ( CF ) due to this genetic mutation . METHODS : A single-center study was performed in CF patients receiving ivacaftor to evaluate the usefulness of high resolution computed tomography ( HRCT ) of the chest as a way to gauge response to ivacaftor therapy . RESULTS : Ten patients with CF were enrolled for at least one year before and after starting ivacaftor . At time of enrollment , mean age was 20.9 ± 10.8 ( range 10-44 ) years . There were significant improvements from baseline to 6 months in mean % FVC ( 93 ± 16 to 99 ± 16 ) and % FEV1 ( 79 ± 26 to 87 ± 28 ) but reverted to baseline at one year . Mean sweat chloride levels decreased significantly from baseline to one year . Mean weight and BMI improved at 6 months . Weight continued to improve with stabilization of BMI at one year . Chest HRCT showed significant improvement at one year in mean modified Brody scores for bronchiectasis , mucous plugging , airway wall thickness , and total Brody scores . Elevated bronchiectasis and airway wall thickness scores correlated significantly with lower % FEV1 , while higher airway wall thickness and mucus plugging scores correlated with more pulmonary exacerbations requiring IV and oral antibiotics respectively . CONCLUSIONS : Based on our findings , HRCT imaging is a useful tool in monitoring response to ivacaftor therapy that corrects the gating defect associated with the G551D- P13569 mutation . Chimeric Q5QGZ9 antibody fusion proteins containing granulocyte-macrophage colony-stimulating factor or interleukin-2 with specificity for B-cell malignancies exhibit enhanced effector functions while retaining tumor targeting properties . Although monoclonal antibody ( MoAb ) therapy of the human malignant lymphomas has shown success in clinical trials , its full potential for the treatment of hematologic malignancies has yet to be realized . To expand the clinical potential of a promising human-mouse chimeric antihuman B-cell MoAb ( chCLL-1 ) constructed using the variable domains cloned from the murine Lym-2 ( muLym-2 ) hybridoma , fusion proteins containing granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) ( chCLL-1/GM- P04141 ) or interleukin ( IL ) -2 ( chCLL-1/ P60568 ) were generated and evaluated for in vitro cytotoxicity and in vivo tumor targeting . The glutamine synthetase gene amplification system was employed for high level expression of the recombinant fusion proteins . Antigenic specificity was confirmed by a competition radioimmunoassay against Q5SW96 -77 human myeloma cells . The activity of chCLL-1/GM- P04141 was established by a colony formation assay , and the bioactivity of chCLL-1/ P60568 was confirmed by supporting the growth of an P60568 -dependent T-cell line . Antibody-dependent cellular cytotoxicity against Q5SW96 -77 target cells demonstrated that both fusion proteins mediate enhanced tumor cell lysis by human mononuclear cells . Finally , biodistribution and imaging studies in nude mice bearing Q5SW96 -77 xenografts indicated that the fusion proteins specifically target the tumors . These in vitro and in vivo data suggest that chCLL-1/GM- P04141 and chCLL-1/ P60568 have potential as immunotherapeutic reagents for the treatment of B-cell malignancies . DB01269 ( vectibix ) . DB01269 ( Vectibix ) , is a human monoclonal antibody P00533 antagonist indicated as a single agent for the treatment of metastatic colorectal carcinoma with disease progression on or following fluoropyrimidine , oxaliplatin , and irinotecan chemotherapy regimens . This article will present the mechanism of action as well as the clinical role for this monoclonal antibody . Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . Antitumor activity of lenvatinib ( e7080 ) : an angiogenesis inhibitor that targets multiple receptor tyrosine kinases in preclinical human thyroid cancer models . Inhibition of tumor angiogenesis by blockading the vascular endothelial growth factor ( P15692 ) signaling pathway is a promising therapeutic strategy for thyroid cancer . DB09078 mesilate ( lenvatinib ) is a potent inhibitor of P15692 receptors ( P17948 -3 ) and other prooncogenic and prooncogenic receptor tyrosine kinases , including fibroblast growth factor receptors ( P11362 -4 ) , platelet derived growth factor receptor α ( PDGFRα ) , P10721 , and P07949 . We examined the antitumor activity of lenvatinib against human thyroid cancer xenograft models in nude mice . Orally administered lenvatinib showed significant antitumor activity in 5 differentiated thyroid cancer ( DTC ) , 5 anaplastic thyroid cancer ( ATC ) , and 1 medullary thyroid cancer ( P04629 ) xenograft models . DB09078 also showed antiangiogenesis activity against 5 DTC and 5 ATC xenografts , while lenvatinib showed in vitro antiproliferative activity against only 2 of 11 thyroid cancer cell lines : that is , RO82-W-1 and TT cells . Western blot analysis showed that cultured RO82-W-1 cells overexpressed P11362 and that lenvatinib inhibited the phosphorylation of P11362 and its downstream effector Q8WU20 . DB09078 also inhibited the phosphorylation of P07949 with the activated mutation C634W in TT cells . These data demonstrate that lenvatinib provides antitumor activity mainly via angiogenesis inhibition but also inhibits FGFR and P07949 signaling pathway in preclinical human thyroid cancer models . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . Synergistic effects of the apolipoprotein E epsilon3/epsilon2/epsilon4 , the cholesteryl ester transfer protein TaqIB , and the apolipoprotein P01024 -482 C > T polymorphisms on their association with coronary artery disease . The single nucleotide polymorphisms ( SNPs ) apolipoprotein E ( P02649 ) epsilon3/epsilon2/epsilon4 , cholesteryl ester transfer protein ( P11597 ) TaqIB , and apolipoprotein P01024 ( P02656 ) -482 C > T have been associated with an atherogenic lipid profile and , in some studies , with increased cardiovascular risk . However , no data exist on their combined impact on atherosclerotic disease . We therefore aimed at investigating the combined impact of these SNPs on the presence of angiographically determined coronary artery disease ( CAD ) . Genotyping was performed in 557 consecutive Caucasian patients undergoing coronary angiography for the evaluation of CAD . From the individual SNPs , only the P02649 epsilon3epsilon4/epsilon4epsilon4 genotype was significantly associated with an increased risk of significant coronary stenoses with lumen narrowing > or=50 % ( odds ratio (OR)=1.77 [ 1.16-2.71 ] ; p=0.008 ) . However , the risk of CAD strongly increased when more than one of the analysed genetic variants was present : ORs were 2.74 [ 1.29-5.83 ] ; p=0.009 for patients with both the P02649 epsilon3epsilon4/epsilon4epsilon4 and the P11597 B1B1 genotype , 1.97 [ 1.06-3.66 ] ; p=0.031 for patients with both the P02649 epsilon3epsilon4/epsilon4epsilon4 genotype and the P02656 -482T allele , 2.12 [ 1.31-3.44 ] ; p=0.002 for patients with both the P11597 B1B1 genotype and the P02656 -482T allele , and 3.99 [ 1.57-13.79 ] ; p=0.029 for patients with all three variants . Multivariate analyses confirmed these results . We conclude that there are strong synergistic effects of the P02649 epsilon3/epsilon2/epsilon4 , the P11597 TaqIB , and the P02656 -482 C > T polymorphisms on their association with CAD . Personalized synthetic lethality induced by targeting P43351 in leukemias identified by gene mutation and expression profile . Homologous recombination repair ( HRR ) protects cells from the lethal effect of spontaneous and therapy-induced DNA double-stand breaks . HRR usually depends on P38398 /2- Q06609 , and P43351 - Q06609 serves as back-up . To target HRR in tumor cells , a phenomenon called " synthetic lethality " was applied , which relies on the addiction of cancer cells to a single DNA repair pathway , whereas normal cells operate 2 or more mechanisms . Using mutagenesis and a peptide aptamer approach , we pinpointed phenylalanine 79 in P43351 DNA binding domain I ( P43351 -phenylalanine 79 [ F79 ] ) as a valid target to induce synthetic lethality in P38398 - and/or P51587 -deficient leukemias and carcinomas without affecting normal cells and tissues . Targeting P43351 -F79 disrupts the P43351 -DNA interaction , resulting in the accumulation of toxic DNA double-stand breaks in malignant cells , but not in normal counterparts . In addition , abrogation of P43351 -DNA interaction enhanced the antileukemia effect of already-approved drugs . BRCA-deficient status predisposing to P43351 -dependent synthetic lethality could be predicted by genetic abnormalities such as oncogenes P11274 - P00519 and P29590 -RAR , mutations in P38398 and/or P51587 genes , and gene expression profiles identifying leukemias displaying low levels of P38398 and/or P51587 . We believe this work may initiate a personalized therapeutic approach in numerous patients with tumors displaying encoded and functional BRCA deficiency . Rapid activation of monocyte tissue factor by antithymocyte globulin is dependent on complement and protein disulfide isomerase . Lymphocyte depletion with antithymocyte globulin ( ATG ) can be complicated by systemic coagulation activation . We found that ATG activated tissue factor procoagulant activity ( TF DB11245 ) on monocytic cells more potently than other stimuli that decrypt TF , including cell disruption , TF pathway inhibitor inhibition , and calcium ionophore treatment . Induction of TF DB11245 by ATG was dependent on lipid raft integrity and complement activation . We showed that ATG-mediated TF activation required complement activation until assembly of the C5b-7 membrane insertion complex , but not lytic pore formation by the membrane attack complex C5b-9 . Consistently , induction of TF DB11245 by ATG did not require maximal phosphatidylserine membrane exposure and was not correlated with the magnitude of complement-induced lytic cell injury . Blockade of free thiols , an inhibitory monoclonal antibody to protein disulfide isomerase ( P07237 ) , and the small-molecule P07237 antagonist DB01698 prevented ATG-mediated TF activation , and P01031 complement activation resulted in oxidation of cell surface P07237 . This rapid and potent mechanism of cellular TF activation represents a novel connection between the complement system and cell surface P07237 -mediated thiol-disulfide exchange . Delineation of this clinically relevant mechanism of activation of the extrinsic coagulation pathway during immunosuppressive therapy with ATG may have broader implications for vascular thrombosis associated with inflammatory disorders . Protein disulphide isomerase is required for signal peptide peptidase-mediated protein degradation . The human cytomegalovirus glycoprotein US2 induces dislocation of MHC class I heavy chains from the endoplasmic reticulum ( ER ) into the cytosol and targets them for proteasomal degradation . Q8TCT9 ( Q8TCT9 ) has been shown to be integral for US2-induced dislocation of MHC class I heavy chains although its mechanism of action remains poorly understood . Here , we show that knockdown of protein disulphide isomerase ( P07237 ) by RNA-mediated interference inhibited the degradation of MHC class I molecules catalysed by US2 but not by its functional homolog US11 . Overexpression of the substrate-binding mutant of P07237 , but not the catalytically inactive mutant , dominant-negatively inhibited US2-mediated dislocation of MHC class I molecules by preventing their release from US2 . Furthermore , P07237 associated with Q8TCT9 independently of US2 and knockdown of P07237 inhibited Q8TCT9 -mediated degradation of CD3delta but not Q9BUN8 -dependent degradation of P13569 DeltaF508 . Together , our data suggest that P07237 is a component of the Q8TCT9 -mediated ER-associated degradation machinery . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Niacin reduces plasma P11597 levels by diminishing liver macrophage content in P11597 transgenic mice . The anti-dyslipidemic drug niacin has recently been shown to reduce the hepatic expression and plasma levels of P11597 . Since liver macrophages contribute to hepatic P11597 expression , we investigated the role of macrophages in the P11597 -lowering effect of niacin in mice . In vitro studies showed that niacin does not directly attenuate P11597 expression in macrophages . Treatment of normolipidemic human P11597 transgenic mice , fed a Western-type diet with niacin for 4 weeks , significantly reduced the hepatic cholesterol concentration ( -20 % ) , hepatic P11597 gene expression ( -20 % ) , and plasma P11597 mass ( -30 % ) . Concomitantly , niacin decreased the hepatic expression of P34810 ( -44 % ) and P45844 ( -32 % ) , both of which are specific markers for the hepatic macrophage content . The decrease in hepatic P11597 expression was significantly correlated with the reduction of hepatic macrophage markers . Furthermore , niacin attenuated atherogenic diet-induced inflammation in liver , as evident from decreased expression of P01375 ( -43 % ) . Niacin similarly decreased the macrophage markers and absolute macrophage content in hyperlipidemic P02649 *3-Leiden. P11597 transgenic mice on a Western-type diet . In conclusion , niacin decreases hepatic P11597 expression and plasma P11597 mass by attenuating liver inflammation and macrophage content in response to its primary lipid-lowering effect , rather than by attenuating the macrophage P11597 expression level . A 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 . P04035 inhibition induces IL-1beta release through Rac1/PI3K/ P31749 -dependent caspase-1 activation . Q03426 deficiency ( MKD ) is an autoinflammatory disorder characterized by recurring fever episodes and results from disturbed isoprenoid biosynthesis . Lipopolysaccharide-stimulated peripheral blood mononuclear cells from MKD patients secrete high levels of interleukin-1beta ( IL-1beta ) because of the presence of hyperactive caspase-1 , and this has been proposed to be the primary cause of recurring inflammation . Here we show that inhibition of P04035 by simvastatin treatment , mimicking MKD , results in increased IL-1beta secretion in a Rac1/PI3K-dependent manner . Simvastatin treatment was found to activate protein kinase B ( P31749 ) /c-akt , a primary effector of PI3K , and ectopic expression of constitutively active P31749 was sufficient to induce IL-1beta release . The small GTPase Rac1 was activated by simvastatin , and this was required for both P31749 activation and IL-1beta secretion . IL-1beta release is mediated by caspase-1 , and simvastatin treatment resulted in increased caspase-1 activity in a Rac1/PI3K-dependent manner . These data suggest that , in MKD , dysregulated isoprenoid biosynthesis activates Rac1/PI3K/ P31749 , resulting in caspase-1 activation with increased IL-1beta release . Importantly , inhibition of Rac1 in peripheral blood mononuclear cells isolated from MKD patients resulted in a dramatic reduction in IL-1beta release . These data suggest that pharmacologic inhibition of Rac1 could provide a novel therapeutic strategy for treatment of MKD . 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 . Translatability scoring in drug development : eight case studies . Translational medicine describes the transfer of basic in vitro and in vivo data into human applications . In the light of low rates of market approvals for new medical entities , better strategies to predict the risk of drug development should be used to increase output and reduce costs . Recently , a scoring system to assess the translatability of early drug projects has been proposed . Here eight drugs from different therapeutic areas have been subjected to a retrospective test-run in this system fictively located at the phase II-III transition . The scores gained here underline the importance of biomarker quality which is pivotal to decrease the risk of the project in all cases . This is particularly evident for gefitinib . The P00533 mutation status is a breakthrough biomarker to predict therapeutic success which made this compound clinically acceptable , and this is plausibly reflected by a considerable increase of the translatability score . For psychiatric and Alzheimer 's drugs , and for a P11597 -inhibitor , the lack of suitable biomarkers and animal models is reflected by a low translatability score , well correlating with the excessive translational risk in these areas . These case studies document the apparent utility of the scoring system , at least under retrospective conditions , as the scores correlate with the outcomes at the level of market approval . Prospective validation is still missing , but these case studies are encouraging . 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 . 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 .

MH_train_1458

MH_train_1458

MH_train_1458

interacts_with DB00215?

multiple_choice

Prolonged lymphocyte depletion by single-dose rabbit anti-thymocyte globulin and alemtuzumab in kidney transplantation . Although antibody induction has gained in popularity , two agents are rarely combined . We retrospectively analyzed peripheral lymphocyte phenotypes of renal transplant recipients who received induction therapy with a different antibody/combination : alemtuzumab(C1H) , DB00098 ( DB00098 ) , daclizumab(Dac) , DB00098 +C1H , and DB00098 +Dac . P01730 + T-cells were suppressed by C1H and DB00098 +C1H , as well as by DB00098 and DB00098 +Dac but to a lesser extent . The effect lasted for 3 years at around 40 % of baseline values . CD8+ T-cells showed a similar trend but had a more rapid recovery to baseline . P15391 + B-cells were effectively suppressed for 2 months by C1H and DB00098 +C1H , and abruptly returned to baseline afterwards ; suppression by DB00098 ( 7 doses ) was modest but lasted longer . A higher proportion of CD56+CD16+ Natural Killer cells in C1H treated patients suggested a relatively spared effect of C1H on this cell type . Low CD25+ T-cells by 5-dose Dac returned to baseline around 6 months , whereas DB00098 +C1H and DB00098 +Dac showed persistent effect . P01730 +CD25hi T-cells were suppressed by both DB00098 +C1H and DB00098 +Dac , but the initial proportion of P01730 +CD25hi T-cells among P01730 + T-cells and P01730 +CD25hi/ P01730 +CD25lo ratio were significantly higher in DB00098 +C1H . Overall , with extensive and persistent lymphocyte suppression by a simple administration of agents , single-dose DB00098 +C1H induction can be an alternative in renal transplantation . 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 . P15391 : A multifunctional immunological target molecule and its implications for Blineage acute lymphoblastic leukemia . Over the last 20-30 years P15391 has gained attention as a potential target in the therapy of B-cell malignancies . In particular , targeting P15391 with the bispecific T-cell engager ( BiTE ) antibody DB09052 and T-cells modified by chimeric antigen receptors ( CAR ) has shown promising efficacy in early phase clinical trials for adults and children with precursor B-cell ALL ( P03999 -ALL ) . This review will discuss the rationale behind targeting P15391 in P03999 -ALL and its potential importance in P03999 -ALL signaling pathways . DB01088 has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 + T cells was measured by ELISA . The expression of the transcription factor FoxP3 after co-culture of DCs with P01730 + CD45RA+ T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 , P05231 , P10145 and IL-12p70 in mDCs , while it enhanced P22301 production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA+ T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP3 expression and P22301 production . Additionally , iloprost inhibited the MIP-3beta-induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans . A novel role for Bruton 's tyrosine kinase in hepatocyte growth factor-mediated immunoregulation of dendritic cells . The limited success of dendritic cell ( DC ) -based immunotherapy in multiple myeloma is partly due to hepatocyte growth factor ( P14210 ) -induced DC dysfunction . From a therapeutic standpoint , it is important to understand the molecular events involved in inhibition of DC activation/maturation by P14210 . Because Bruton 's tyrosine kinase ( Btk ) negatively regulates maturation and immunostimulatory function of DCs , a role for Btk in P14210 -induced inhibition of both murine and human DCs was investigated . We demonstrate that Btk is a novel proximal component of P14210 -induced c-MET ( P08581 ) signaling . Following P14210 treatment , Btk binds to c-MET and becomes activated . Btk activation in turn blocks the NF-κB pathway and subsequent DC activation via the c-Src-PI3K-AKT-mammalian target of rapamycin ( P42345 ) pathway . Notably , Btk activation is necessary for P14210 -induced association of c-Src and PI3K with c-MET . Furthermore , we provide the first evidence that P14210 inhibits DC activation by inducing autocrine interleukin ( IL ) -10 secretion , which requires activation of Btk . Blocking activation of Btk and its downstream the c-Src-PI3K-AKT- P42345 pathway prevents P14210 -induced P22301 secretion by DCs . In addition , neutralization of P22301 secretion from DCs impaired the inhibitory effect of P14210 on DCs . Thus , our study identifies a novel role for Btk in P14210 -induced DC inhibition . 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 . 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 . 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 . DB05101 and cetuximab activate the epidermal growth factor receptor but fail to trigger downstream signaling by Akt or Erk . Molecular inhibition of the epidermal growth factor receptor ( P00533 ) is a promising anticancer strategy , and monoclonal antibodies ( mAbs ) to P00533 are undergoing extensive evaluation in preclinical and clinical trials . However , the effects of anti- P00533 mAbs on P00533 signaling have remained unclear . We have now examined the effects of 2 anti- P00533 mAbs , matuzumab ( EMD72000 ) and cetuximab ( Erbitux ) , both of which are currently under assessment for treatment of various cancers , on P00533 signal transduction and cell survival in nonsmall cell lung cancer cell lines . Similar to P01133 , matuzumab and cetuximab each induced phosphorylation of P00533 at several tyrosine phosphorylation sites as a result of receptor dimerization and activation of the receptor tyrosine kinase . In contrast to the effects of P01133 , however , P00533 activation induced by these antibodies was not accompanied by receptor turnover or by activation of downstream signaling pathways that are mediated by Akt and Erk and are important for regulation of cell proliferation and survival . In addition , clonogenic survival assays revealed that matuzumab and cetuximab reduced the survival rate of H292 cells , in which they also inhibited the P01133 -induced activation of Akt and Erk . Although we have examined only a few cell lines , our results indicate that the antitumor effects of matuzumab and cetuximab depend on inhibition of P00533 downstream signaling mediated by Akt or Erk rather than on inhibition of P00533 itself . Vitamin D up-regulates the vitamin D receptor by protecting it from proteasomal degradation in human P01730 + T cells . The active form of vitamin D3 , 1,25(OH)2D3 , has significant immunomodulatory properties and is an important determinant in the differentiation of P01730 + effector T cells . The biological actions of 1,25(OH)2D3 are mediated by the vitamin D receptor ( P11473 ) and are believed to correlate with the P11473 protein expression level in a given cell . The aim of this study was to determine if and how 1,25(OH)2D3 by itself regulates P11473 expression in human P01730 + T cells . We found that activated P01730 + T cells have the capacity to convert the inactive DB00146 to the active 1,25(OH)2D3 that subsequently up-regulates P11473 protein expression approximately 2-fold . 1,25(OH)2D3 does not increase P11473 mRNA expression but increases the half-life of the P11473 protein in activated P01730 + T cells . Furthermore , 1,25(OH)2D3 induces a significant intracellular redistribution of the P11473 . We show that 1,25(OH)2D3 stabilizes the P11473 by protecting it from proteasomal degradation . Finally , we demonstrate that proteasome inhibition leads to up-regulation of P11473 protein expression and increases 1,25(OH)2D3-induced gene activation . In conclusion , our study shows that activated P01730 + T cells can produce 1,25(OH)2D3 , and that 1,25(OH)2D3 induces a 2-fold up-regulation of the P11473 protein expression in activated P01730 + T cells by protecting the P11473 against proteasomal degradation . 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 . 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 . 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 . 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 . Management of ocular inflammation and pain following cataract surgery : focus on bromfenac ophthalmic solution . Recently , several new ophthalmic NSAID products have been introduced for commercial use in the United States . The purpose of this review is to briefly overview the ophthalmic NSAIDs currently in use and to discuss the management of postoperative ocular inflammation and pain following cataract surgery with a particular focus on bromfenac ophthalmic solution 0.09 % . DB00963 ophthalmic solution 0.09 % is indicated for the reduction of ocular pain and inflammation following cataract surgery . Studies have shown that bromfenac ophthalmic solution 0.09 % has equivalent efficacy to the other topical NSAIDs in reducing postsurgical inflammation and controlling pain . The unique chemical structure of bromfenac makes it both a potent inhibitor of the P35354 enzyme and a highly lipophilic molecule that rapidly penetrates to produce early and sustained drug levels in all ocular tissues . Clinically , these pharmacokinetic features are manifested in a rapid reduction of postsurgical inflammation and pain with bid dosing . DB00963 ophthalmic solution 0.09 % is a versatile agent and is effective when used as either monotherapy or as an adjunct therapy to steroids . In vitro cell death of activated lymphocytes in Omenn 's syndrome . Omenn 's syndrome ( OS ) is characterized by erythrodermia , hepatosplenomegaly , lymphadenopathy , hypereosinophilia and elevated IgE levels associated with increased susceptibility to severe infections . Peripheral blood T cells , though usually present in normal number , show an activated phenotype ( including an increased expression of CD95/Fas ) , a Th2 pattern of cytokine secretion and defective proliferative response to mitogens . In this report , we demonstrate that T cells from patients with OS undergo an excessive cell death in vitro resulting from two mechanisms . First , a substantial number of peripheral blood lymphocytes from OS patients die in unstimulated cultures ( p = 0.009 vs. healthy controls ) . This spontaneous apoptosis is associated with reduced expression of bcl-2 gene product ( p < 0.05 ) and can be prevented by addition of interleukin ( IL ) -2 ( which also prevents down-modulation of bcl-2 ) , while is independent from CD95 signaling . Second , lymphocytes from OS patients are highly susceptible to activation-induced cell death ( AICD ) induced with mitogens . This mechanism is largely independent from P60568 , while it can be significantly inhibited blocking CD95 with an IgG2b monoclonal antibody ( mAb ) . The dependence of AICD from signals transduced via CD95 was confirmed showing that cross-linking CD95 with an IgM mAb induces a higher cell death in purified P01730 + CD45R0+ cells from OS patients than in controls ( comparable for CD95 expression ) . Both mechanisms of cell death observed in this study result from lymphocyte hyperactivation occurring in vivo in these patients and may contribute to functional T cell defects of OS . 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 . Local immunotherapy of glioma patients with a combination of 2 bispecific antibody fragments and resting autologous lymphocytes : evidence for in situ t-cell activation and therapeutic efficacy . After adoptive transfer of pre-activated lymphocytes into the operation cavity of glioma patients , tumor regression and improved survival have been reported in some patients . Results were most impressive when bispecific antibodies with tumor x CD3 specificity were also applied . In this study , we attempted to avoid time-consuming pre-activation procedures for adoptively transferred cells by using a combination of bispecific antibodies directed to the P01133 receptor ( P00533 ) on tumor cells and to CD3 and P10747 on T cells . Eleven patients with high-grade malignant glioma received 3 injections of 2 bispecific antibody fragments ( P00533 x CD3 and P00533 x P10747 ) together with freshly isolated autologous lymphocytes via an Ommaya reservoir . Intracavitary fluid aspirated during immunotherapy was examined for markers of T-cell activation . Increased levels of soluble P60568 receptor and P01375 were detected in the intracavitary fluid of all patients tested . Two of the 11 treated patients experienced a beneficial response to therapy as defined by a transient contrast enhancement in subsequent Q9BWK5 scans and prolonged survival . Side effects were transient and consisted of fever , nausea , headache and aggravation of pre-existing neurologic deficits . These adverse effects were most likely due to the antibody construct containing anti-CD3 specificity . Two patients developed cerebral edema and required steroid treatment . 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 .

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interacts_with DB03880?

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Metabolic fate of 2,2-dimethylbutyryl moiety of simvastatin in rats : identification of metabolites by gas chromatography/mass spectrometry . Metabolic pathways of simvastatin ( DB00641 ) , a lactone prodrug of an inhibitor of P04035 , were elucidated in male rats , using the [ 14C ] -labelled compound . Evidence has been obtained for hydrolysis of simvastatin and its metabolites at their 2,2-dimethylbutyryl moieties . Metabolites identified in plasma were 2,2-dimethylbutyric acid ( P28068 ) , 2,2-dimethyl-3-hydroxybutyric acid ( DMHB ) and an open chain hydroxy acid of simvastatin : metabolites identified in urine were DMHB , a glucuronide and the glycine conjugate of P28068 . They were characterized by gas chromatography/electron impact and chemical ionization mass spectrometry as phenacyl or pertrimethylsilylated derivatives . The structures of the metabolites and the aglycone of the glucuronide were confirmed as phenacyl esters by comparison of their chromatographic data and mass spectra with those of the phenacyl derivatives of authentic compounds . T lymphocytes induce endothelial cell matrix metalloproteinase expression by a P29965 -dependent mechanism : implications for tubule formation . Neovascularization frequently accompanies chronic immune responses characterized by T cell infiltration and activation . Angiogenesis requires endothelial cells ( ECs ) to penetrate extracellular matrix , a process that involves matrix metalloproteinases ( MMPs ) . We report here that activated human T cells mediate contact-dependent expression of MMPs in ECs through P25942 / P29965 signaling . Ligation of P25942 on ECs induced de novo expression of gelatinase B ( P14780 ) , increased interstitial collagenase ( P03956 ) and stromelysin ( P08254 ) , and activated gelatinase A ( P08253 ) . Recombinant human P29965 induced expression of MMPs by human vascular ECs to a greater extent than did maximally effective concentrations of interleukin-1beta or tumor necrosis factor-alpha . Moreover , activation of human vascular ECs through P25942 induced tube formation in a three-dimensional fibrin matrix gel assay , an effect antagonized by a MMP inhibitor . These results demonstrated that activation of ECs by interaction with T cells induced synthesis and release of MMPs and promoted an angiogenic function of ECs via P29965 - P25942 signaling . As vascular cells at the sites of chronic inflammation , such as atherosclerotic plaques , express P25942 and its ligand , our findings suggest that ligation of P25942 on ECs can mediate aspects of vascular remodeling and neovessel formation during atherogenesis and other chronic immune reactions . 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 . The correlation between difference in foreign body reaction between implant locations and cytokine and MMP expression . The foreign body reaction ( FBR ) differs between subcutaneously and supra-epicardially implanted materials . We hypothesize that this is a result of differences in cytokine , chemokine and matrix metalloproteinase ( MMP ) dynamics . Therefore we applied collagen disks subcutaneously and on the epicardium in mice and analyzed the FBR from day 1 to 21 . Both the influx of leukocytes and implant degradation were higher in supra-epicardially implanted collagen than in subcutaneously implanted material . This correlated with a higher gene expression of pro-inflammatory cytokines such as IL-1 and P05231 , and a lower expression of the anti-inflammatory cytokine P22301 . Furthermore , the higher supra-epicardial expression of PMN attractants P09341 /KC and P19875 / Q9H7D7 correlated with a higher and prolonged PMN influx . The gene expression levels of collagen degrading MMPs , i.e. P22894 , P45452 and P50281 were similar in subcutaneous and supra-epicardial disks . However , the activity of these enzymes was markedly higher supra-epicardially . In addition , the P14780 expression was higher supra-epicardially , suggesting a role for this enzyme in the degradation process . In conclusion , a strong pro-inflammatory milieu is generated after supra-epicardial implantation that enables prolonged PMN presence and activation . This , together with the high supra-epicardial P14780 level , could explain the observed difference in Col-I degradation between locations . Pathologic indicators of degradation and inflammation in human osteoarthritic cartilage are abrogated by exposure to n-3 fatty acids . OBJECTIVE : To determine if n-3 polyunsaturated fatty acid ( PUFA ) supplementation ( versus treatment with n-6 polyunsaturated or other fatty acid supplements ) affects the metabolism of osteoarthritic ( OA ) cartilage . METHODS : The metabolic profile of human OA cartilage was determined at the time of harvest and after 24-hour exposure to n-3 PUFAs or other classes of fatty acids , followed by explant culture for 4 days in the presence or absence of interleukin-1 ( IL-1 ) . Parameters measured were glycosaminoglycan release , aggrecanase and matrix metalloproteinase ( MMP ) activity , and the levels of expression of messenger RNA ( mRNA ) for mediators of inflammation , aggrecanases , MMPs , and their natural tissue inhibitors ( tissue inhibitors of metalloproteinases [ TIMPs ] ) . RESULTS : Supplementation with n-3 PUFA ( but not other fatty acids ) reduced , in a dose-dependent manner , the endogenous and IL-1-induced release of proteoglycan metabolites from articular cartilage explants and specifically abolished endogenous aggrecanase and collagenase proteolytic activity . Similarly , expression of mRNA for O75173 , P45452 , and P08254 ( but not P01033 , -2 , or -3 ) was also specifically abolished with n-3 PUFA supplementation . In addition , n-3 PUFA supplementation abolished the expression of mRNA for mediators of inflammation ( cyclooxygenase 2 , P09917 , P09917 -activating protein , tumor necrosis factor alpha , IL-1alpha , and IL-1beta ) without affecting the expression of message for several other proteins involved in normal tissue homeostasis . CONCLUSION : These studies show that the pathologic indicators manifested in human OA cartilage can be significantly altered by exposure of the cartilage to n-3 PUFA , but not to other classes of fatty acids . 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 . Chronic induction . What 's new in the pipeline . Induction therapy with biological agents was introduced the in the 1970s and the rationale , concepts and approach have remained almost unchanged for 30 years . However , the novel biological agents being developed for induction therapy are being designed for chronic rather than short-term therapy with several objectives : reduce dependence on toxic and nephrotoxic agents , improve outcome and ultimately facilitate the emergence of tolerance . The biological agents include efalizumab , a humanized anti-CD11a ( anti-LFA1 ) , anti- CD154 , anti- P25942 , a number of agents targeting P40933 and its receptor , and costimulation blockade with humanized antibodies to P33681 / P42081 and the fusion receptor protein DB06681 , a second generation DB01281 . The past decade has witnessed an unprecedented number of small molecules/oral drugs that have been developed and approved for renal transplantation ; the next decade , however , may witness the emergence of a new class of biological induction agents that may displace some of the currently used drugs . 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 . 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 . 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 . Expression of vitamin D receptor and metabolizing enzymes in multiple sclerosis-affected brain tissue . Vitamin D deficiency has been implicated as a risk factor for multiple sclerosis ( MS ) , but how vitamin D metabolism affects MS pathophysiology is not understood . We studied the expression of vitamin D receptor ( P11473 ) and related enzymes , including 1,25(OH)(2)D-24-hydroxylase ( Q07973 ; Q07973 ) and 25(OH)D-1α-hydroxylase ( O15528 ) , in CNS tissues of 39 MS patients and 20 controls and in primary human glial cells in vitro . In control and MS normal-appearing white matter ( NAWM ) , nuclear P11473 immunostaining was observed in oligodendrocyte-like cells , human leukocyte antigen ( HLA ) -positive microglia , and glial fibrillary acidic protein-positive astrocytes . There was a 2-fold increase in P11473 transcripts in MS NAWM versus control white matter ( p = 0.03 ) . In chronic active MS lesions , HLA-positive microglia/macrophages showed nuclear P11473 staining ; astrocytes showed nuclear and cytoplasmic P11473 staining . Staining for Q07973 was restricted to astrocytes. P11473 and O15528 mRNA expressions were increased in active MS lesions versus NAWM ( p < 0.01 , p = 0.04 , respectively ) . In primary human astrocytes in vitro , the active form of vitamin D , 1,25(OH)(2)D(3) , induced upregulation of P11473 and Q07973 . P01375 and interferon-γ upregulated O15528 mRNA in primary human microglia and astrocytes . Increased P11473 expression in MS NAWM and inflammatory cytokine-induced amplified expression of P11473 and O15528 in chronic active MS lesions suggest increased sensitivity to vitamin D in NAWM and a possible endogenous role for vitamin D metabolism in the suppression of active MS lesions . 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 . 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 . 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 . 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 . Spondyloarthritides . The most important clinical features of the spondyloarthritides ( SpA ) are not only inflammatory back pain ( Q9H4E7 ) but also peripheral ( enthesitis ) and extra-articular symptoms . For clinical purposes , two forms related to the predominant clinical manifestation - axial and peripheral SpA - and five subgroups- ankylosing spondylitis ( AS ) , SpA associated with psoriasis and inflammatory bowel disease ( Q9UKU7 ) , reactive arthritis and undifferentiated SpA - are differentiated . Axial SpA including AS is the most frequent subtype of SpA , followed by psoriatic arthritis and undifferentiated SpA , while reactive arthritis and Q9UKU7 -related SpA are less frequent . The prevalence of SpA has been shown to be similar to rheumatoid arthritis . The outcome of the disease is influenced by the degree of disease activity over time , which is mainly related not only to inflammation but also on the structural damage ( new bone formation ) that occurs over time . Treatment options for patients with SpA have been limited for decades . Non-steroidal anti-inflammatory agents are currently considered first choice , since they have shown good amelioration of symptoms in SpA patients especially when suffering by the typical symptom of Q9H4E7 . Furthermore , there is a clear role for regular physiotherapy in AS to prevent loss of spinal mobility . For patients who have insufficiently responded to conventional therapies , four anti-tumour necrosis factor ( P01375 ) agents are available and are approved for the treatment of patients with active AS : infliximab , etanercept , adalimumab and DB06674 . As far as it stands now , P01375 blockers seem to have no influence on new bone formation in AS . Genetic influences on sarcoidosis . To investigate the genetic influences underlying the development of sarcoidosis , HLA class II genotyping was performed in Japanese patients with sarcoidosis and healthy controls using the PCR-RFLP method . The frequencies of both DR52 group antigen-associated alleles ( HLA- Q8IUH3 *11 , - Q8IUH3 *12 and - Q8IUH3 *14 ) and Q8IUH3 *08 alleles were higher in the patient group , suggesting that the common , specific amino acid residue on the Q8IUH3 molecule of these alleles may determine susceptibility to sarcoidosis . Alternatively , it is possible that another susceptibility gene , linked to these Q8IUH3 alleles , exists within the MHC region . We screened the P01375 , P01374 , P0DMV8 and Hum70t genes around the class III region , as well as the P28067 and - P28068 genes in the class II region , for genetic polymorphism in sarcoidosis . None of these genes suggested a susceptibility to sarcoidosis . These studies support the thesis that one of the major genetic factors controlling the development of sarcoidosis is located within the Q8IUH3 locus in the HLA class II region . 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 .

MH_train_1460

MH_train_1460

MH_train_1460

interacts_with DB00623?

multiple_choice

[ 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 . Simultaneous inhibition of MEK and P11802 leads to potent apoptosis in human melanoma cells . ABSTRACT Deregulation of DB01367 -RAF-MEK- P29323 and P42771 -cycylin D: P11802 /6-RB pathways is important for melanoma development . Chemotherapeutic agents targeting both pathways were developed but results of clinical studies with monotherapies were disappointing . We examined the effect of co-targeting both pathways with MEK inhibitor PD98059 and P11802 inhibitor 219476 on human melanoma cells lines , and found that combinatorial treatment dramatically increased apoptosis compared to the single agent treatment . The apoptosis was associated with downregulation of P10415 , Q07817 , O15392 , and upregulation of O43521 . Our results indicate that simultaneously targeting P29323 and RB pathways is a promising strategy for melanoma treatment and should encourage further in-depth investigations . 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 ) 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 . Polymorphism identification in the P11310 , P01008 , P22301 , P15173 and P01222 genes of cattle . DY-9760e inhibits endothelin-1-induced cardiomyocyte hypertrophy through inhibition of CaMKII and P29323 activities . Ca(2+)/calmodulin-dependent protein kinase II ( CaMKII ) and extracellular signal-regulated kinase ( P29323 ) have pivotal roles in endothelin-1 ( ET-1 ) -induced cardiomyocyte hypertrophy . We here tested whether a novel P62158 antagonist , DY-9760e inhibits ET-1-induced hypertrophy through inhibition of CaMKII and P29323 activities . We first confirmed that Ca(2+) oscillation induced by ET-1 treatment elicits transient activation of CaMKII and P29323 in cultured cardiomyocytes . DY-9760e treatment with 3 microM totally and partially inhibited the ET-1-induced CaMKII and P29323 activation , respectively . The ET-1-induced P29323 activation was also partially blocked by a CaMKII inhibitor , KN93 . To confirm involvement of CaMKII activity in the P29323 activation by ET-1 and A23187 , cultured cardiomyocytes were transfected with a constitutively active CaMKII . The transfection with the active CaMKII elicited P29323 activation in cultured cardiomyocytes and cotransfection with dominant negative CaMKII eliminated its P29323 activation . Consistent with inhibitory actions of DY-9760e on the ET-1-induced CaMKII and P29323 activation , induction of hypertrophy-related genes including atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) was significantly inhibited by DY-9760e treatment . Combination treatment with DY-9760e and U0126 , a MEK inhibitor , totally blocked the ET-1-induced P01160 and DB04899 expression . DY-9760e treatment ( 3 microM ) significantly inhibited the ET-1-induced hypertrophy and combination treatment with DY-9760e and U0126 totally blocked the ET-1-induced hypertrophy in cultured cardiomyocytes . These results suggest that DY-9760e elicits antihypertrophic action on ET-1-induced cardiac hypertrophy through inhibition of CaMKII and P29323 activation and that CaMKII activity in part mediates ET-1-induced P29323 activation . 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 . DB01373 -induced folding of a fragment of calmodulin composed of EF-hands 2 and 3 . P62158 ( P62158 ) is an EF-hand protein composed of two calcium ( Ca(2+) ) -binding EF-hand motifs in its N-domain ( EF-1 and P13639 ) and two in its C-domain ( EF-3 and Q8N442 ) . In this study , we examined the structure , dynamics , and Ca(2+)-binding properties of a fragment of P62158 containing only P13639 and EF-3 and the intervening linker sequence ( CaM2/3 ) . Based on NMR spectroscopic analyses , Ca(2+)-free CaM2/3 is predominantly unfolded , but upon binding Ca(2+) , adopts a monomeric structure composed of two EF-hand motifs bridged by a short antiparallel beta-sheet . Despite having an " even-odd " pairing of EF-hands , the tertiary structure of CaM2/3 is similar to both the " odd-even " paired N- and C-domains of Ca(2+)-ligated P62158 , with the conformationally flexible linker sequence adopting the role of an inter-EF-hand loop . However , unlike either P62158 domain , CaM2/3 exhibits stepwise Ca(2+) binding with a K ( d1 ) = 30 +/- 5 microM to EF-3 , and a K ( d2 ) > 1000 microM to P13639 . Binding of the first equivalent of Ca(2+) induces the cooperative folding of CaM2/3 . In the case of native P62158 , stacking interactions between four conserved aromatic residues help to hold the first and fourth helices of each EF-hand domain together , while the loop between EF-hands covalently tethers the second and third helices . In contrast , these aromatic residues lie along the second and third helices of CaM2/3 , and thus are positioned adjacent to the loop between its " even-odd " paired EF-hands . This nonnative hydrophobic core packing may contribute to the weak Ca(2+) affinity exhibited by P13639 in the context of CaM2/3 . 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 . Production of an antiserum specific to the ADP-ribosylated form of elongation factor 2 from archaebacteria and eukaryotes . An antiserum to ADP-ribosylated elongation factor 2 ( DB02059 - P13639 ) from S. acidocaldarius was raised in rabbits using stained , homogenized , DB02059 - P13639 -containing slices from SDS-gels as a source of antigen . P13639 ( P13639 ) from S. acidocaldarius was cloned in E. coli and the expressed gene product was used in order to adsorb all anti- P13639 antibodies which do not contain the ADP-ribosyl group within their epitopes , as E. coli is unable to synthesize the ADP-ribosyl acceptor diphthamide . The remaining antibodies were specific to ADP-ribosylated P13639 from Thermoplasma acidophilum , S. acidocaldarius and Desulfurococcus mucosus . ADP-ribosylated P13639 from eukaryotic sources also reacted with the adsorbed antiserum as shown for P13639 isolated from the killi-fish Cynolebias whitei , the mouse species BALB/c and Han/Wistar rats . The adsorbed antiserum did not cross-react with ADP-ribosylated actin or rho protein or with DB03147 -containing D-amino acid oxidase . Platelet-associated antibodies , cellular immunity and FCGR3a genotype influence the response to rituximab in immune thrombocytopenia . DB00073 is widely used in autoimmune diseases including immune thrombocytopenia ( ITP ) , although the mechanism of effect remains unclear . This study describes the effects of rituximab on platelet-associated antibodies ( PA-APAs ) , B and T cell counts and clonality ( IGHV and TRG @ gene rearrangements ) , P08637 ( FcγRIIIa ) and P12318 ( FcγRIIa ) polymorphisms and correlation to anti- P29965 ( P29965 ) response . PA-APA levels fell more frequently in responders ( 6/8 ) than in non-responders ( 2/10 : P = 0·08-0·15 ) . Two responders had no PA-APAs . Two non-responders with a fall in PA-APAs had very high CD8 levels . One non-responder had a B cell clone , one responder and one non-responder had a T cell clone . 15/16 patients had the same responses to rituximab and antiCD40L . Patients with P08637 V/V polymorphisms were more likely to respond to rituximab ( P = 0·03 ) . In summary , the fall in PA-APAs in responders confirms the humoural effect of rituximab . Failure to respond in patients with very high CD8 levels , despite PA-APA fall indicates a role for T cell-mediated platelet/megakaryocyte destruction . Concordance of response to anti- P29965 suggests autoantibody-producing cells are under T cell control . Finally , the effect of FCGR polymorphisms on response confirms the importance of FCGR-mediated depletion of B cells in autoimmunity . This has implications on the pathology of ITP as well as the immunological effect of B cell depletion . DB06096 , a selective P29475 inhibitor and a P28222 /1D receptor agonist , inhibits P80511 release in preclinical migraine models . BACKGROUND : DB06096 is a combined neuronal nitric oxide synthase ( P29475 ) inhibitor and 5-hydroxytryptamine 1B/1D ( P28222 /1D ) receptor agonist . Using preclinical models , we evaluated whether these two unique therapeutic principles have a synergistic effect in attenuating stimulated calcitonin gene-related peptide ( P80511 ) release , a marker of trigeminal activation . METHODS : We examined the effect of DB06096 on : ( 1 ) DB00761 - , capsaicin- and resiniferatoxin ( RTX ) -induced immunoreactive P80511 ( iCGRP ) release from isolated preparation of rat dura mater , trigeminal ganglion ( TG ) and trigeminal nucleus caudalis ( P24821 ) ; and ( 2 ) capsaicin- and electrical stimulation ( ES ) -induced middle meningeal artery ( MMA ) dilation in a rat closed-cranial window . RESULTS : DB06096 inhibited : ( 1 ) DB00761 -stimulated iCGRP release from dura mater ( % decrease mean ± SEM , lowest effective concentration ) ( 35 ± 6 % , 30 µM ) , TG ( 24 ± 11 % , 10 µM ) and P24821 ( 40 ± 8 % , 10 µM ) ; ( 2 ) capsaicin- and RTX-induced iCGRP release from dura mater ; and ( 3 ) capsaicin- and ES-induced increase in dural artery diameter ( 32 ± 5 % , 3 mg kg(-1) intravenous ( i.v. ) and 36 ± 1 % , 10 mg kg(-1) i.v. ) . CONCLUSIONS : DB06096 inhibits P80511 release from migraine-relevant cephalic tissues . Its effect is most likely mediated via a combination of P29475 -inhibition and P28222 /1D receptor agonism in dura mater while the mechanisms of action for inhibition of P80511 release from TG and P24821 have to be investigated further . 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 . 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 . 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 . 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 . 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 . 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 . Red wine extract protects against oxidative-stress-induced endothelial senescence . Red wine polyphenols may preserve endothelial function during aging . Endothelial cell senescence enhances age-related endothelial dysfunction . We investigated whether RWE ( red wine extract ) prevents oxidative-stress-induced senescence in HUVECs ( human umbilical-vein endothelial cells ) . Senescence was induced by exposing HUVECs to tBHP ( t-butylhydroperoxide ) , and quantified by senescence-associated β-galactosidase staining . RWE ( 0-50 μg/ml ) concentration dependently decreased senescence by maximally 33±7.1 % . RWE prevented the senescence-associated increase in P38936 protein expression , inhibited tBHP-induced DNA damage of endothelial cells and induced relaxation of PCAs ( porcine coronary arteries ) . Inhibition of Q96EB6 ( sirtuin 1 ) by sirtinol partially reversed the effect of RWE on tBHP-induced senescence , whereas both the NOS ( nitric oxide synthase ) inhibitor L-NMMA ( NG-monomethyl-L-arginine ) and the P36551 ( cyclo-oxygenase ) inhibitor indomethacin fully inhibited it . Furthermore , incubation of HUVECs with RWE increased P29474 ( endothelial NOS ) and P35354 mRNA levels as well as phosphorylation of P29474 at Ser1177 . RWE protects endothelial cells from tBHP-induced senescence . NO and P35354 , in addition to activation of Q96EB6 , play a critical role in the inhibition of senescence induction in human endothelial cells by RWE . Increased levels of Candida albicans mannan-specific T-cell-derived antigen binding molecules in patients with invasive candidiasis . In addition to cytokines , P01730 + T cells have been found to secrete soluble , T-cell-derived antigen binding molecules ( TABMs ) . These antigen-specific immunoproteins are thought to have immunoregulatory properties in the suppression of cell-mediated immunity ( CMI ) because they often associate with interleukin-10 ( P22301 ) and transforming growth factor beta . Decreased CMI causes susceptibility to infections caused by organisms which are normally nonpathogenic . In this situation , e.g. , Candida albicans saprophytism may develop into invasive candidiasis . The difficult diagnosis of invasive candidiasis is based on the findings obtained from blood cultures and with tissue biopsy specimens , with some additional diagnostic value gained by the detection of Candida albicans mannan antigenemia and antimannan antibodies . In the present study , Candida albicans mannan-specific TABM ( P62158 -TABM ) levels in the sera of patients with invasive candidiasis ( n = 11 ) , Candida colonization ( n = 11 ) and noncolonization ( n = 10 ) , recurrent vulvovaginal candidiasis ( n = 30 ) , and atopic eczema dermatitis syndrome ( n = 59 ) and healthy controls ( n = 30 ) were analyzed . For 14 participants , the effect of mannan stimulation on TABM production and gamma interferon ( P01579 ) and P05112 mRNA expression by peripheral blood lymphocytes was also studied . It was demonstrated that P62158 -TABM production was the highest in patients with invasive candidiasis and that P62158 -TABM levels could distinguish Candida-colonized patients from noncolonized patients . In addition , the P62158 -TABM level was directly related to mRNA expression for P05112 but not P01579 . These results reinforce the view that TABMs are associated with decreased CMI , immunoregulation , and the T-helper cell 2-type immune response . Platelets trigger a P25942 -dependent inflammatory response in the microvasculature of inflammatory bowel disease patients . BACKGROUND & AIMS : Platelets circulate in an activated state in patients with inflammatory bowel disease ( Q9UKU7 ) , but their role in the pathogenesis of Q9UKU7 is unclear . The recent demonstration that activated platelets express P29965 ( L ) provides a mechanism of interaction with P25942 -positive endothelial cells , inducing them to produce proinflammatory mediators . We investigated whether platelets from patients with Q9UKU7 express enhanced levels of P29965 and induce human intestinal microvascular endothelial cells ( HIMEC ) to up-regulate cell adhesion molecule ( P62158 ) expression and secrete chemokines . METHODS : P29965 expression was assessed in resting and thrombin-activated platelets by flow cytometry and in mucosal microthrombi by confocal microscopy . Platelet-HIMEC cocultures were used to study P62158 up-regulation , and interleukin ( IL ) -8 and RANTES production by HIMEC . RESULTS : Q9UKU7 platelets expressed significantly higher P29965 levels than those of healthy subjects , and P29965 -positive platelets were detected in Q9UKU7 -involved mucosa . Activated platelets up-regulated expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 as well as production of interleukin 8 by HIMEC in a P25942 -dependent fashion . High levels of RANTES were present in platelet-HIMEC cocultures and platelets were identified as the source of this chemokine , which mediated T-cell adhesion to HIMEC . CONCLUSIONS : These results show that platelets can actively contribute to mucosal inflammation and represent a previously unrecognized component of Q9UKU7 pathogenesis . 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 .

MH_train_1461

MH_train_1461

MH_train_1461

interacts_with DB00015?

multiple_choice

A bispecific antifibrin-antiplatelet urokinase conjugate ( BAAUC ) induces enhanced clot lysis and inhibits platelet aggregation . Thrombolysis is well established in the treatment of acute myocardial infarction . However , clinical application of thrombolytic agents has limitations with respect to efficacy and specificity . To achieve highly effective and at the same time clot-selective plasminogen activation urokinase was coupled to a bispecific antibody consisting of the monovalent Fab ' from the antifibrin monoclonal antibody 59D8 and the monovalent Fab ' from the anti-glycoprotein P08514 /IIIa monoclonal antibody DB00054 . The bispecific antifibrin-antiplatelet urokinase conjugate ( BAAUC ) was synthesized and characterized . Assays with either immobilized platelets , P08514 /IIIa or fibrin showed an increase in plasminogen activation compared to uncoupled urokinase by 10-fold , 58-fold and 13-fold , respectivley ( p < 0.0001 each ) . In vitro clot lysis was performed on platelet-rich and fibrin-rich clots and revealed an up to 5-fold higher potency of BAAUC compared to uncoupled urokinase ( p < 0.0001 ) . In vitro platelet aggregation was effectively inhibited by the hybrid molecule , whereas urokinase had no effect . BAAUC and two monospecific urokinase-conjugates , UK-59D8-IgG and UK-7E3-(Fab')2 were compared with each other with regard to similar tests . In vitro clot assays with platelet-rich and platelet-poor clots were performed . BAAUC achieved a significantly higher plasminogen activation compared to each of the monospecific conjugates ( p < 0.05 , respectively ) . We conclude that BAAUC , a bispecific plasminogen activator with antifibrin and antiplatelet properties has the potency to lyse both fibrin-rich and platelet-rich thrombi with high efficacy and to effectively inhibit platelet aggregation . 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 . [ DB02424 administration reduces the number of P07900 -positive germ cells in the mouse embryo : preliminary results ] . 5 mg of DB02424 , an inhibitor of stress protein P07900 which express on mammalian germ cells , were administered to E8 pregnant mice . E17 embryos were removed , and a quantitative analysis of HSP90-immunoreactive cells in the gonad was performed , in comparison to control embryos . First , we observed that the number of germ cells is lower in male than in female embryos , as well in control and experimental embryos . External features of experimental and control embryos did not display any difference . Embryos exposed to geldanamycin exhibit a significant decrease of immunoreactive germ cells . In two embryos , we observed a group of ectopic immunoreactive cells in the pelvic area . We conclude that geldanamycin inhibits germ cells migration , and suggest that this inhibition can lead to ectopic germ cell populations , similar to teratomas . Deciphering the systems biology of P42345 inhibition by integrative transcriptome analysis . The P42345 signaling plays an integral role in cellular homeostasis controlling the transition between the catabolic and anabolic states . Originally approved as immunosuppressive agents preventing allograft rejection , inhibitors of P42345 signaling have recently entered the arena of cancer therapy . Using rapamycin derivative ( RAD001 ) as a prototype inhibitor , we aimed to systematically analyze the molecular mechanisms underlying the pleiotropic effects of P42345 signaling . Using proliferation- and clonogenic survival assays , a preferential sensitivity of microvascular endothelial cells ( HDMVEC ) followed by fibroblasts and U87 gliblastoma to RAD001 treatment was found . In contrast , lung- and prostate tumor cells demonstrated relative resistance against RAD001 treatment . In co-culture with fibroblasts , RAD001 exerted potent antiangiogenic effects by inhibiting endothelial cell tube formation . Further , RAD001 treatment efficiently prevented tumor growth in U87 tumor xenografts . Integrative transcriptome analysis was performed to decipher the molecular mechanism underlying RAD001 -induced anti-tumor and antiangiogenic effects . The predominant expression pattern was downregulation of genes after RAD001 treatment in all three sensitive cell types . Among the RAD001 downregulated genes , a transcriptional network was discovered enriched for genes related to angiogenesis processes and extracellular matrix remodeling , e.g. , P15692 , Q16665 , CXCLs , P05231 , FN , P05121 or NRP1 . Of note , key components of PI3K upstream ( PDK1 ) as well as mTORC2 downstream signaling ( O00141 , NDRG ) were downregulated by RAD001 . Decreased expression of IMPDH and 139 common gene targets between mycophenolic acid and RAD001 suggested in part shared mechanisms underlying their antiangiogenic and immunosuppressive effects . In summary , key genetic participants governing anti-tumor and anti-angiogenic effects of P42345 inhibition were identified . 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 . 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 . 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 . Pocket protein-independent repression of urokinase-type plasminogen activator and plasminogen activator inhibitor 1 gene expression by Q01094 . Expression of genes of the plasminogen activator ( PA ) system declines at the G(0)/G(1)-S-phase boundary of the cell cycle . We found that overexpression of Q01094 -3 , which acts mainly in late G(1) , inhibits promoter activity and endogenous expression of the urokinase-type PA ( uPA ) and PA inhibitor 1 ( P05121 ) genes . This effect is dose dependent and conserved in evolution . Mutation analysis indicated that both the DNA-binding and transactivation domains of Q01094 are necessary for this regulation . Interestingly , an Q01094 mutant lacking the pRB-binding region strongly repressed the uPA and P05121 promoters . An E2F-mediated negative effect was also observed in pRB and P28749 / Q08999 knockout cell lines . This is the first report that E2F can act as a repressor independently of pocket proteins . Mutation of AP-1 elements in the uPA promoter abrogated E2F-mediated transcriptional inhibition , suggesting the involvement of AP-1 in this regulation . Results shown here identify E2F as an important component of transcriptional control of the PA system and thus provide new insights into mechanisms of cellular proliferation . 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 . 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 . Nuclear and cytoplasmic interaction of Q08999 / Q08999 and Q92731 in MCF-7 breast cancer cells . DB00286 exhibit important biological functions and influence several pathological processes of hormone-dependent diseases . The biological actions of estrogens require their interaction with two estrogen receptors ( P03372 and Q92731 ) , which are ligand-dependent transcription factors . P03372 and Q92731 exhibit distinct tissue expression patterns as well as show different patterns of gene regulation . In addition , it has been suggested that Q92731 works as a counter partner of P03372 through inhibition of the transactivating functions of P03372 . For instance , Q92731 seems to play a different role in breast tumorigenesis than P03372 , as Q92731 decreased expression in breast cancer has been correlated with bad prognosis . Biological activities of P03372 and Q92731 could be controlled by a number of interacting proteins such as activators/inhibitors , ligand binding and kinases . We have previously reported that Q08999 / Q08999 , retinoblastoma related protein , could be involved in the silencing of P03372 gene during breast tumorigenesis . Here , we report that Q92731 and Q08999 / Q08999 proteins co-immunoprecipitate in both nucleus and cytoplasm of MCF-7 breast cancer cells . Our hypothesis is that the interaction of Q08999 /130 with Q92731 may have a functional significance in regulating Q92731 activity . The Pro/Pro genotype of the p53 codon 72 polymorphism modulates P05121 plasma levels in ageing . P05121 ( P05121 ) is over-expressed during ageing and it has been linked to cellular senescence . Recently , P05121 has been also identified in vitro as a critical downstream target of p53 . P04637 , the p53 gene , has a common functional polymorphism at codon 72 which influences the capability to modulate both apoptosis and cell senescence . In the attempt to demonstrate an in vivo role of p53 in the relationship between P05121 and age , we studied P05121 on 570 healthy subjects ( aged from 18 to 92yrs. ) . P05121 showed significant relationship with age ( r=0.12 , p=0.02 ) . Stratifying by genotype , it became evident that the association between P05121 and age was mainly due to Pro/Pro subjects ( partial r=0.75 , p < 0.01 ) . These results have been confirmed by a validation study on an independent sample population of 496 subjects ( aged from 18 to 94yrs. ) . This is the first demonstration of an in vivo role of P04637 polymorphism in P05121 regulation , supporting the hypothesis that the effects of this polymorphism are age-dependent . In particular , our results indicate that Pro/Pro genotype plays a pivotal role in determining P05121 levels in aged subjects , while in DB00125 carriers P05121 levels are associated to the known insulin related determinants . 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 . 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 . Q9BYW2 -α and survivin involved in the anti-apoptotic effect of DB02342 after global ischemia in rats . Survivin is an anti-apoptotic gene that decreases the apoptosis by depressing the expression of caspase-3 . Hypoxia-inducible factor-1-alpha ( Q16665 ) is a transcription factor specifically activated by hypoxia . 2-methoxyestradiol ( DB02342 ) is an estradiol derivative and a known Q16665 inhibitor . DB02342 decreased apoptosis by inhibiting Q16665 . The aim of the present study was to investigate if survivin is involved in the anti-apoptotic effect of DB02342 . Male adult rats were used to make the global ischemia ( GI ) model . Ten minutes after GI , DB02342 was injected intraperitoneally ( 16 mg/kg weight ) . Rats were killed at 6 hours , 12 hours , 24 hours , 48 hours , 96 hours , and 7 days . GI produced a marked increase in Q16665 expressions in the hippocampus at 6 hours and peaked at 48-96 hours . The expressions of survivin and caspase-3 were increased lightly in a similar time course . These molecular changes were accompanied by massive cell loss and apoptosis in the hippocampal regions . DB02342 treatment reduced the expression of Q16665 , increased survivin expression , and decreased the expression of caspase-3 . These results indicate that survivin and Q16665 were involved in the anti-apoptotic effect of DB02342 treated following GI . DB02342 may decrease the Q16665 expression , up-regulate the survivin expression , inhibit the expression of caspase-3 , and finally reduce apoptosis after GI . 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 . Transcription factor Q9Y261 -centered transcriptional regulation network in non-small cell lung cancer . Lung cancer is the leading cause of cancer-mediated death . Although various therapeutic approaches are used for lung cancer treatment , these mainly target the tumor suppressor p53 transcription factor , which is involved in apoptosis and cell cycle arrest . However , p53-targeted therapies have limited application in lung cancer , since p53 is found to be mutated in more than half of lung cancers . In this study , we propose tumor suppressor Q9Y261 as an alternative target protein for therapies against lung cancer and reveal a possible Q9Y261 -centered transcriptional regulation network by identifying new target genes and binding partners of Q9Y261 by using various screening techniques . The genes encoding DB00142 / DB00128 -rich carboxy-terminal domain 2 ( Q99967 ) , nuclear receptor subfamily 0 , group B , member 2 ( Q15466 ) , cell adhesion molecule 1 ( Q9BY67 ) and P10415 -associated X protein ( Q07812 ) were identified as putative target genes of Q9Y261 . Additionally , the proteins including highly similar to heat shock protein P08238 -beta ( P07900 ) , heat shock 70 kDa protein 1A variant ( P0DMV8 ) , histone deacetylase 1 ( Q13547 ) and O15379 were identified as novel interacting partners of Q9Y261 . Moreover , we showed that Q9Y261 -dependent promoter activation of Q07812 and P38936 genes is significantly reduced via physical interactions between the identified binding partners and Q9Y261 . These results provide opportunities to understand the Q9Y261 -centered transcriptional regulation network and novel therapeutic targets to modulate this network in p53-deficient lung cancer . Secretome profiling reveals the signaling molecules of apoptotic HCT116 cells induced by the dietary polyacetylene gymnasterkoreayne B . Dietary polyacetylenes from various foods have been receiving attention as promising cancer chemopreventive agents . However , until now , the detailed molecular mechanism and the regulatory proteins underlying these effects have not been elucidated . We investigated the effects of gymnasterkoreayne B ( GKB ) , a model dietary polyacetylene from wild vegetables , on the programmed cell death of HCT116 human colorectal cancer cells . GKB inhibited HCT116 cell proliferation by inducing apoptotic cell death . GKB treatment resulted in ROS accumulation , leading to the activation of both intrinsic and extrinsic apoptotic pathway . We also found that P02751 , P01137 , P05067 , P05121 , P10809 , P00441 , P10599 , and O43707 may act as secretory signaling molecules during GKB-induced apoptotic cell death using LC-MS/MS identification followed by spectrum counting , statistical calculation , and gene ontology analysis . The secretory proteins suggested in this study may be promising candidates involved in apoptotic cell death of cancer cells induced by GKB that warrant further functional study . Gluco- and mineralocorticoid receptor-mediated regulation of neurotrophic factor gene expression in the dorsal hippocampus and the neocortex of the rat . Gluco- and mineralocorticoid receptors ( GR and MR ) act via common promoter elements but may exert different effects on gene regulation in various regions of the forebrain . In order to separately analyse the role of GR and MR in the regulation of neurotrophic factor genes and their receptors , we used adrenalectomy and subsequent hormone injections in the rat as a model system . Twenty-four hours after adrenalectomy rats were injected with a single dose of corticosterone ( 2 and 10 mg/kg ) , aldosterone ( 0.5 mg/kg ) or the synthetic glucocorticoid agonist RU 28362 ( 4 mg/kg ) . Gene expression of basic fibroblast growth factor ( P09038 ) and its high-affinity receptors [ fibroblast growth factor receptor subtypes 1-3 ( FGF- Q96GN5 , FGF-R2 , FGF-R3 ) ] , as well as brain-derived growth factor ( P23560 ) and neurotrophin-3 ( P20783 ) was analysed at 4 h after the hormone injection in P00915 - P22748 ( cornus of Ammon areas of the hippocampus ) and dentate gyrus of the dorsal hippocampus and in neocortex by means of in situ hybridization . We found that P09038 is regulated in P00918 , P07451 and dentate gyrus by GR and MR together , and in P00915 , P22748 and neocortex by GR alone . FGF-R2 expression in the hippocampus seems to be regulated only by MR , while P23560 expression appears to depend on both receptors . FGF- Q96GN5 , FGF-R3 and P20783 were only moderately affected by the hormone activation of GR and MR acting in concert or alone in the various regions . Thus , the present findings suggest that the adrenal cortical system through GR and MR participate in the control of neurotrophic factor signalling in a highly subregion- and cellular-dependent manner . Iptakalim enhances adult mouse hippocampal neurogenesis via opening Kir6.1-composed K- DB00171 channels expressed in neural stem cells . BACKGROUND AND PURPOSE : Emerging evidence indicates that stimulating adult neurogenesis provides novel strategies for central nervous system diseases . Iptakalim ( Ipt ) , a novel DB00171 -sensitive potassium ( K- DB00171 ) channel opener , has been demonstrated to play multipotential neuroprotective effects in vivo and in vitro . However , it remains unknown whether Ipt could regulate the adult neurogenesis . METHODS AND RESULTS : Based on the finding that adult neural stem cells ( ANSCs ) in hippocampus expressed Kir6.1/ Q09428 -composed K- DB00171 channel , Kir6.1 heterozygotic ( Kir6.1(+/-) ) mice were used to investigate whether and how Ipt regulates adult hippocampal neurogenesis . We showed that administration of Ipt ( 10 mg/kg ) or fluoxetine ( Flx , 10 mg/kg ) for 4 weeks significantly increased newborn ANSCs in subgranular zone ( SGZ ) of Kir6.1(+/+) mice but failed to affect those of Kir6.1(+/-) mice . Meanwhile , ANSCs in Kir6.1(+/-) mice exhibited decreased survival rate and impaired ability of differentiation into astrocytes . We further found that Kir6.1(+/-) mice showed lower level of brain-derived neurotrophic factor ( P23560 ) in hippocampus compared with Kir6.1(+/+) mice . Furthermore , Ipt increased the levels of P23560 and basic fibroblast growth factor ( P09038 ) throughout the hippocampus in Kir6.1(+/+) mice but not in Kir6.1(+/-) mice . Moreover , Ipt and Flx enhanced the phosphorylation of Akt and CREB in the hippocampus of Kir6.1(+/+) mice . Notably , these effects were completely abolished in Kir6.1(+/-) mice . CONCLUSIONS : Our findings demonstrate that Ipt stimulates the adult hippocampal neurogenesis via activation of Akt and CREB signal following the opening of Kir6.1-composed K- DB00171 channels , which gives us an insight into the therapeutic implication of Ipt in the diseases with adult neurogenesis deficiency , such as major depression .

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interacts_with DB09073?

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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 . 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 . [ Expression of human papillomavirus and P06400 in head and neck squamous cell carcinoma ] . OBJECTIVE : The purpose of this study was to examine the interrelationship between human papilloma virus ( HPV ) and retinoblastoma protein ( P06400 ) in patients with squamous cell carcinoma of the head and neck ( SCCHN ) . METHOD : Archival material from 33 oropharynx , 40 oral cavity was selected for study based on the availability of tissue from the primary tumors prior to surgery . HPV detection was done using general primer P40197 (+) Q9HCN6 (+) mediated PCR enzyme immunoassay ( PCR-EIA ) and typed with type-specific probes . Monoclonal antibodies specific for P06400 was used for immunohistochemical detection of Rb protein in these primary tumors . RESULT : HPV DNA was detected in 12.3 % of tumors . All were HPV 16 DNA . When stratified , 18.0 % of oropharyngeal cancers was positive compared with 7.5 % of oral cavity tumors ; P06400 negative was 12.3 % . HPV positive patients tended to present with a higher stage and lymph node metastasis . In spite of the higher stage at presentation , survival in patients with HPV positive and P06400 negative SCCHN was significantly longer than that of HPV-negative patients . CONCLUSION : Patients with HPV-positive tumors may have a survival advantage relative to patients with HPV-negative tumors . This indicates that HPV-positive and P06400 -negative tumors are sensitive to radiotherapy . [ DB08836 treatment upregulated cardiomyocyte thioredoxin expression and improved autoimmune myocarditis ] . 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 : The up-regulation of TRX by temocapril treatment was checked by Western blot in normal rat myocytes in vitro and in vivo , as well as in rats with experimental autoimmune myocarditis ( EAM ) . RESULTS : DB08836 enhanced cytosolic redox regulatory protein TRX expression , but 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 EAM , the severity of myocarditis and the protein carbonyl contents were less increased in temocapril treatment ( 10 mg x kg(-1) x d(-1) , orally ) from day 1 to day 21 , but not in temocapril treatment from day 15 to day 21 . If the characteristics of this model that myocardial inflammation begins around day 15 and keeps on until day 21 is considered , temocapril treatment for 3 weeks might be thought as a preconditioning treatment . CONCLUSIONS : TRX and the redox state modified by TRX may play a crucial role in the pathophysiology of EAM . DB08836 ameliorates myocarditis with inducing TRX up-regulation in a preconditioning manner , although the mechanism of TRX up-regulation by temocapril remains to be elucidated . 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 . Lipopolysaccharide-induced dopaminergic cell death in rat midbrain slice cultures : role of inducible nitric oxide synthase and protection by indomethacin . Glial cell activation associated with inflammatory reaction may contribute to pathogenic processes of neurodegenerative disorders , through production of several cytotoxic molecules . We investigated the consequences of glial activation by interferon-gamma ( P01579 ) /lipopolysaccharide ( LPS ) in rat midbrain slice cultures . Application of P01579 followed by LPS caused dopaminergic cell death and accompanying increases in nitrite production and lactate dehydrogenase release . DB02533 , an inhibitor of inducible nitric oxide synthase ( P35228 ) , or SB203580 , an inhibitor of p38 mitogen-activated protein kinase , prevented dopaminergic cell loss as well as nitrite production . SB203580 also suppressed expression of P35228 and cyclooxygenase-2 ( P35354 ) induced by P01579 /LPS . A P36551 inhibitor indomethacin protected dopaminergic neurons from P01579 /LPS-induced injury , whereas selective P35354 inhibitors such as NS-398 and nimesulide did not . Notably , indomethacin was able to attenuate neurotoxicity of a nitric oxide ( NO ) donor . Neutralizing antibodies against tumour necrosis factor-alpha and interleukin-1beta did not inhibit dopaminergic cell death caused by P01579 /LPS , although combined application of these antibodies blocked lactate dehydrogenase release and decrease in the number of non-dopaminergic neurons . These results indicate that P35228 -derived NO plays a crucial role in P01579 /LPS-induced dopaminergic cell death , and that indomethacin exerts protective effect by mechanisms probably related to NO neurotoxicity rather than through P36551 inhibition . 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 . Endothelial cells negatively modulate reactive oxygen species generation in vascular smooth muscle cells : role of thioredoxin . In intact vessels , endothelial cells ( ECs ) and vascular smooth muscle cells ( VSMCs ) act as an integrated system , possibly through reactive oxygen species ( ROS ) . Using a coculture system we tested whether ECs modulate VSMC redox status by regulating activity of NAD(P)H oxidase and antioxidants . VSMC production of O(2)(*-) , H(2)O(2) , and NO was assessed using fluoroprobes and amplex-red . NAD(P)H oxidase subunit expression and oxidase activity were determined by Western blotting and chemiluminescence , respectively . Expression of thioredoxin , SOD , growth signaling pathways ( P12004 , p21cip1 , P11802 , P27361 /2 , p38MAPK ) was evaluated by immunoblotting . P10599 activity was assessed by the insulin disulfide reduction assay . In cocultured conditions , VSMC ROS production was reduced by approximately 50 % without changes in NAD(P)H oxidase expression/activity versus monoculture ( P < 0.05 ) . This was associated with decreased cell growth ( P < 0.05 ) . Expression of Cu/Zn SOD and thioredoxin was increased in coculture versus monoculture VSMCs ( P < 0.01 ) . Pretreatment of ECs with L-NAME ( NOS inhibitor ) , NS-398 ( Cox2 inhibitor ) , and HET0016 ( 20-HETE inhibitor ) did not influence VSMC ROS formation , whereas CDNB , thioredoxin reductase inhibitor , abolished ROS modulating effects of ECs . These findings indicate that in a coculture system recapitulating intact vessels , ECs negatively regulate ROS production in VSMCs through thioredoxin upregulation . Functionally this is associated with growth inhibition . The modulatory actions of ECs are independent of NOS/NO , Cox2 , and HETE and do not involve NAD(P)H oxidase . Our data identify novel mechanisms whereby ECs protect against VSMC oxidative stress , a process that may be important in maintaining vascular integrity . 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 . Hedyotis diffusa Willd inhibits colorectal cancer growth in vivo via inhibition of P40763 signaling pathway . Signal Transducer and Activator of Transcription 3 ( P40763 ) , a common oncogenic mediator , is constitutively activated in many types of human cancers ; therefore it is a major focus in the development of novel anti-cancer agents . Hedyotis diffusa Willd has been used as a major component in several Chinese medicine formulas for the clinical treatment of colorectal cancer ( CRC ) . However , the precise mechanism of its anti-tumor activity remains largely unclear . Using a CRC mouse xenograft model , in the present study we evaluated the effect of the ethanol extract of Hedyotis diffusa Willd ( EEHDW ) on tumor growth in vivo and investigated the underlying molecular mechanisms . We found that EEHDW reduced tumor volume and tumor weight , but had no effect on body weight gain in CRC mice , demonstrating that EEHDW can inhibit CRC growth in vivo without apparent adverse effect . In addition , EEHDW treatment suppressed P40763 phosphorylation in tumor tissues , which in turn resulted in the promotion of cancer cell apoptosis and inhibition of proliferation . Moreover , EEHDW treatment altered the expression pattern of several important target genes of the P40763 signaling pathway , i.e. , decreased expression of P12004 D1 , P11802 and Bcl-2 as well as up-regulated P38936 and Bax . These results suggest that suppression of the P40763 pathway might be one of the mechanisms by which EEHDW treats colorectal cancer . [ Genetic risk factors of neural tube defects ] . Neural tube defects ( NTDs ) are a group of diseases caused by a failure of closure of the neural tube . Its aetiology contains both environmental and genetic factors . NTDs have a polygenic background . Genes , which are linked with NTDs occurrence , are both directly and indirectly connected with controlling the process of closure of the neural tube . Ones of those are genes of metabolism of folic acid as P42898 , Q99707 , Q9UBK8 , P35520 , P11586 , folic acid receptors ( FR ) regulator genes from PAX family , T , P16234 and P38398 genes . 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 . 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 . Cytotoxic effects of fascaplysin against small cell lung cancer cell lines . Fascaplysin , the natural product of a marine sponge , exhibits anticancer activity against a broad range of tumor cells , presumably through interaction with DNA , and/or as a highly selective cyclin-dependent kinase 4 ( P11802 ) inhibitor . In this study , cytotoxic activity of fascaplysin against a panel of small cell lung cancer ( SCLC ) cell lines and putative synergism with chemotherapeutics was investigated . SCLC responds to first-line chemotherapy with platinum-based drugs/etoposide , but relapses early with topotecan remaining as the single approved therapeutic agent . Fascaplysin was found to show high cytotoxicity against SCLC cells and to induce cell cycle arrest in P55008 /0 at lower and S-phase at higher concentrations , respectively . The compound generated reactive oxygen species ( ROS ) and induced apoptotic cell death in the chemoresistant NCI-H417 SCLC cell line . Furthermore , fascaplysin revealed marked synergism with the topoisomerase I-directed camptothecin and 10-hydroxy-camptothecin . The Poly(ADP-ribose)-Polymerase 1 ( P09874 ) inhibitor Q06418 204165 antagonized the cytotoxic activity of fascaplysin , pointing to the involvement of DNA repair in response to the anticancer activity of the drug . In conclusion , fascaplysin seems to be suitable for treatment of SCLC , based on high cytotoxic activity through multiple routes of action , affecting topoisomerase I , integrity of DNA and generation of ROS . JTE-522 , a selective P35354 inhibitor , inhibits cell proliferation and induces apoptosis in RL95-2 cells . AIM : To investigate whether JTE-522 [ 4-(4-cyclohexyl-2-methyloxazol-5-yl)-2-fluorobenzenesulfonamide ] , a selective P35354 inhibitor , can induce apoptosis and inhibit cell proliferation in human endometrial cancer cell line RL95-2 cells and to explore the molecular mechanisms . METHODS : [ 3-(4,5)-dimethylthiazol-2-yl ] -2,5-diphenyl tetrazolium bromide ( MTT ) , DNA ladder , enzyme-linked immunosorbent assay ( ELISA ) , flow cytometry , RT-PCR , and Western blot analysis were employed to investigate effect of JTE-522 on human endometrial cancer cell line RL95-2 cells and the related molecular mechanisms . RESULTS : JTE-522 inhibited the growth of RL95-2 cells and induced the apoptosis . Furthermore , it arrested G0/ P55008 phase and inhibited S phase in RL95-2 cells . JTE-522 inhibited the expressions of P35354 mRNA , phosphorylated Rb , and P11802 proteins , while increased the levels of p53 , P38936 , cyclin D1 proteins , and the activity of caspase-3 in RL95-2 cells . CONCLUSION : JTE-522 inhibits cell proliferation and induces apoptosis in RL95-2 cells , which may be associated with the activation of caspase-3-like proteases , down-regulation of the expression of P35354 mRNA , phosphorylated Rb , and P11802 proteins , and up-regulation of the expressions of p53 , P38936 , and cyclin D1 proteins . DB09073 ( PD 0332991 ) : targeting the cell cycle machinery in breast cancer . INTRODUCTION : The cyclin D-cyclin-dependent kinases 4 and 6 ( P11802 /6 ) -retinoblastoma ( P06400 ) pathway , governing the cell cycle restriction point , is frequently altered in breast cancer and is a potentially relevant target for anticancer therapy . DB09073 ( PD 0332991 ) , a potent and selective inhibitor of P11802 and Q00534 , inhibits proliferation of several P06400 -positive cancer cell lines and xenograft models . AREAS COVERED : The basic features and abnormalities of the cell cycle in breast cancer are described , along with their involvement in estrogen signaling and endocrine resistance . The pharmacological features of palbociclib , its activity in preclinical models of breast cancer and the potential determinants of response are then illustrated , and its clinical development in breast cancer described . A literature search on the topic was conducted through PubMed and the proceedings of the main cancer congresses of recent years . EXPERT OPINION : The combination of palbociclib with endocrine agents is a very promising treatment and Phase III clinical trials are ongoing to confirm its efficacy . Further , potentially useful combinations are those with drugs targeting mitogenic signaling pathways , such as P04626 - and PI3K-inhibitors . Combination with chemotherapy seems more problematic , as antagonism has been reported in preclinical models . The identification of predictive factors , already explored in preclinical studies , must be further refined and validated in clinical trials . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used . 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 . Gene expression profiling in response to the histone deacetylase inhibitor BL1521 in neuroblastoma . Neuroblastoma is a childhood tumor with a poor survival in advanced stage disease despite intensive chemotherapeutic regimes . The new histone deacetylase ( HDAC ) inhibitor BL1521 has shown promising results in neuroblastoma . Inhibition of HDAC resulted in a decrease in proliferation and metabolic activity , induction of apoptosis and differentiation of neuroblastoma cells . In order to elucidate the mechanism mediating the effects of BL1521 on neuroblastoma cells , we investigated the gene expression profile of an P04198 single copy ( SKNAS ) and an P04198 amplified ( IMR32 ) neuroblastoma cell line after treatment with BL1521 using the Affymetrix oligonucleotide array U133A . An altered expression of 255 genes was observed in both neuroblastoma cell lines . The majority of these genes were involved in gene expression , cellular metabolism , and cell signaling . We observed changes in the expression of vital genes belonging to the cell cycle ( cyclin D1 and P11802 ) and apoptosis ( Q12983 , P55957 , and P10415 ) pathway in response to BL1521 . The expression of 37 genes was altered by both BL1521 and Trichostatin A , which could indicate a common gene set regulated by different HDAC inhibitors . BL1521 treatment changed the expression of a number of P04198 -associated genes . Several genes in the Wnt and the Delta/Notch pathways were changed in response to BL1521 treatment , suggesting that BL1521 is able to induce the differentiation of neuroblastoma cells into a more mature phenotype . 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 . 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 .

MH_train_1463

MH_train_1463

MH_train_1463

interacts_with DB01114?

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 . Pharmacodynamics and pharmacokinetics of DB05225 , a novel inhibitor of P09917 -activating protein ( P20292 ) . The P09917 -activating protein ( P20292 ) gene and an increase in leukotriene ( LT ) production are linked to the risk of asthma , myocardial infarction , and stroke . We evaluated the pharmacodynamics , pharmacokinetics , and tolerability of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel P20292 inhibitor , in healthy subjects . Single and multiple doses of DB05225 demonstrated dose-dependent inhibition of blood Q06643 (4) production and dose-related inhibition of urinary LTE(4) . After a single oral dose ( 50-1,000 mg ) of DB05225 , the maximum concentration ( C(max) ) and area under the curve ( AUC ) in plasma increased in a dose-dependent manner . After multiple-dose administration ( 50-1,000 mg once daily for 11 days ) , there were no significant differences in the pharmacokinetic parameters between the first and last days of treatment . DB05225 was well tolerated at all doses in both the single- and multiple-dose cohorts . Further clinical trials with DB05225 in inflammatory diseases are warranted . DB11320 inhibits adrenocortical cell proliferation but does not affect steroidogenesis . DB11320 ( HA ) is a neurotransmitter synthesized in most mammalian tissues exclusively by histidine decarboxylase enzyme . Among the plethora of actions mediated by HA , the modulatory effects on steroidogenesis and proliferation in Leydig cells ( LCs ) have been described recently . To determine whether the effects on LCs reported could be extrapolated to all steroidogenic systems , in this study , we assessed the effect of this amine on adrenal proliferation and steroidogenesis , using two adrenocortical cell lines as experimental models , murine Q03519 cells and human NCI-H295R cells . Even when steroidogenesis was not modified by HA in adrenocortical cells , the biogenic amine inhibited the proliferation of H295R cells . This action was mediated by the activation of P35367 subtype and an increase in the production of inositol phosphates as second messengers , causing cell-cycle arrest in the G2/M phase . These results indicate a new role for HA in the proliferation of human adrenocortical cells that could contribute to a better understanding of tumor pathology as well as to the development of new therapeutic agents . 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 . P18827 and angiogenic cytokines in multiple myeloma : correlation with bone marrow angiogenesis and survival . Angiogenesis is a complex process involved in the proliferation and metastasis of malignant tumours , and partly triggered by the secretion of various angiogenic factors by tumour cells or cells in the stromal environment . We investigated the correlation between bone marrow angiogenesis , estimated as microvessel density ( P53602 ) , and interleukin-6 ( P05231 ) , basic fibroblastic growth factor ( P09038 ) , hepatocyte growth factor ( P14210 ) and syndecan-1 in 67 patients with newly diagnosed multiple myeloma , and evaluated the prognostic value of these parameters . Circulating levels of P05231 , P09038 , P14210 and syndecan-1 were significantly higher in patients than in controls . Moreover , in patients , bone marrow levels of P09038 , P14210 and syndecan-1 were higher than peripheral blood levels . Positive correlations were found between P53602 and syndecan-1 blood levels ( r = 0.33 , P = 0.017 ) , syndecan-1 bone marrow levels ( r = 0.49 , P = 0.046 ) and P14210 blood levels ( r = 0.36 , P = 0.008 ) respectively . High P53602 and high blood levels of P05231 , P14210 and syndecan-1 were predictive of a shorter survival . In a multivariate survival analysis P53602 and blood levels of P05231 retained independent prognostic significance , while in a survival analysis without P53602 the peripheral blood levels of P14210 and syndecan-1 were strong independent prognostic factors . [ 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 . 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 . Dynamic stereochemistry of Topiramate ( anticonvulsant drug ) in solution : theoretical approaches and experimental validation . Topiramate , an antiepileptic drug , was synthesized with an improved protocol and identified by (1)H NMR , (13)C NMR , (1)H-(1)H COSY , HMQC and HMBC spectrum . In parallel , density functional theory ( DFT ) using B3LYP functional and split-valance 6-311++G** basis set has been used to optimize the structures and conformers of Topiramate . Also experimental and theoretical methods have been used to correlate the dependencies of (1)J and (2)J involving (1)H and (13)C on the C1- P06681 ( ω ) and C1-O1 ( θ ) torsion angles in the glycosidic part of Topiramate . New Karplus equations are proposed to assist in the structural interpretation of these couplings . Importantly , due to the sensitivity of some couplings , most notably (2)J( P35367 ,H1S) , (2)J( P06681 , P35367 ) and (2)J( P06681 ,H1S) values depend on both C-C ( ω ) and C-O ( θ ) torsion angles . Analyses of experimental coupling constants for protons on the pyranose ring of Topiramate indicate a twist boat structure for Topiramate in solution . In all calculations solvent effects were considered using a polarized continuum model ( PCM ) . 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 . 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 . Follicular dendritic cells in health and disease . Follicular dendritic cells ( FDCs ) are unique immune cells that contribute to the regulation of humoral immune responses . These cells are located in the B-cell follicles of secondary lymphoid tissues where they trap and retain antigens ( Ags ) in the form of highly immunogenic immune complexes ( ICs ) consisting of Ag plus specific antibody ( Ab ) and/or complement proteins . FDCs multimerize Ags and present them polyvalently to B-cells in periodically arranged arrays that extensively crosslink the B-cell receptors for Ag ( BCRs ) . P18627 -FcγRIIB mediates IC periodicity , and P18627 -Ag presentation combined with other soluble and membrane bound signals contributed by FDCs , like P18627 - Q9Y275 , - P05231 , and -C4bBP , are essential for the induction of the germinal center ( GC ) reaction , the maintenance of serological memory , and the remarkable ability of P18627 -Ags to induce specific Ab responses in the absence of cognate T-cell help . On the other hand , FDCs play a negative role in several disease conditions including chronic inflammatory diseases , autoimmune diseases , HIV/AIDS , prion diseases , and follicular lymphomas . Compared to other accessory immune cells , FDCs have received little attention , and their functions have not been fully elucidated . This review gives an overview of P18627 structure , and recapitulates our current knowledge on the immunoregulatory functions of FDCs in health and disease . A better understanding of FDCs should permit better regulation of Ab responses to suit the therapeutic manipulation of regulated and dysregulated immune responses . Type B gamma-aminobutyric acid receptors modulate the function of the extracellular Ca2+-sensing receptor and cell differentiation in murine growth plate chondrocytes . Extracellular calcium-sensing receptors ( CaRs ) and metabotropic or type B gamma-aminobutyric acid receptors ( GABA-B-Rs ) , two closely related members of family C of the G protein-coupled receptor superfamily , dimerize in the formation of signaling and membrane-anchored receptor complexes . We tested whether CaRs and two GABA-B-R subunits ( Q96GN5 and R2 ) are expressed in mouse growth plate chondrocytes ( GPCs ) by PCR and immunocytochemistry and whether interactions between these receptors influence the expression and function of the CaR and extracellular Ca(2+)-mediated cell differentiation . Both CaRs and the Q9UBS5 and -R2 were expressed in the same zones of the growth plate and extensively colocalized in intracellular compartments and on the membranes of cultured GPCs . The Q9UBS5 co-immunoprecipitated with the CaR , confirming a physical interaction between the two receptors in GPCs . In vitro knockout of Q9UBS5 genes , using a Cre-lox recombination strategy , blunted the ability of high extracellular Ca(2+) concentration to activate phospholipase C and P27361 /2 , suppressed cell proliferation , and enhanced apoptosis in cultured GPCs . In GPCs , in which the Q9UBS5 was acutely knocked down , there was reduced expression of early chondrocyte markers , aggrecan and type II collagen , and increased expression of the late differentiation markers , type X collagen and osteopontin . These results support the idea that physical interactions between CaRs and GABA-B-R1s modulate the growth and differentiation of GPCs , potentially by altering the function of CaRs . 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 . 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 . 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 . B cell-targeted therapies for systemic lupus erythematosus : an update on clinical trial data . In the past year there has been remarkable activity and some important success in the development of B cell-targeted therapies for the treatment of systemic lupus erythematosus ( SLE ) . The most promising studies were BLISS-52 and BLISS-76 , large phase III studies that demonstrated measurable efficacy for belimumab , a monoclonal antibody against B cell-activating factor ( Q9Y275 ) . The moderate-sized phase II/III trials EXPLORER and LUNAR that tested rituximab , an anti- P11836 monoclonal antibody , for treatment of non-renal and renal lupus , disappointed many investigators with anecdotal success in refractory patients . These rituximab trials were intended to detect a large clinical effect in patients with very active disease and this was not found . Nevertheless , arguments can be made for additional studies in targeted populations or with a change in design to detect smaller or longer-term effects . DB04958 , a monoclonal antibody against the B cell surface antigen P20273 , and atacicept , a chimeric molecule formed by a receptor for Q9Y275 and a proliferation-inducing ligand ( APRIL ) with immunoglobulin ( Ig ) -G , have both been promising in initial small trials and now larger clinical trials are underway . Thus , recent clinical trial data show that B cell-targeting therapies are beginning to fulfil their promise as treatments for SLE and there are good reasons to hope for further progress in the near future . [ 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 . Glucagon-like peptide-1 inhibits adipose tissue macrophage infiltration and inflammation in an obese mouse model of diabetes . AIMS/HYPOTHESIS : Obesity and insulin resistance are associated with low-grade chronic inflammation . Glucagon-like peptide-1 ( P0C6A0 ) is known to reduce insulin resistance . We investigated whether P0C6A0 has anti-inflammatory effects on adipose tissue , including adipocytes and adipose tissue macrophages ( Q13315 ) . METHODS : We administered a recombinant adenovirus ( rAd ) producing P0C6A0 ( rAd- P0C6A0 ) to an ob/ob mouse model of diabetes . We examined insulin sensitivity , body fat mass , the infiltration of Q13315 and metabolic profiles . We analysed the mRNA expression of inflammatory cytokines , lipogenic genes , and M1 and M2 macrophage-specific genes in adipose tissue by real-time quantitative PCR . We also examined the activation of nuclear factor κB ( NF-κB ) , extracellular signal-regulated kinase 1/2 and Jun N-terminal kinase ( JNK ) in vivo and in vitro . RESULTS : Fat mass , adipocyte size and mRNA expression of lipogenic genes were significantly reduced in adipose tissue of rAd- P0C6A0 -treated ob/ob mice . Macrophage populations ( F4/80(+) and F4/80(+)CD11b(+)CD11c(+) cells ) , as well as the expression and production of P05231 , P01375 -α and monocyte chemoattractant protein-1 , were significantly reduced in adipose tissue of rAd- P0C6A0 -treated ob/ob mice . Expression of M1-specific mRNAs was significantly reduced , but that of M2-specific mRNAs was unchanged in rAd- P0C6A0 -treated ob/ob mice . NF-κB and JNK activation was significantly reduced in adipose tissue of rAd- P0C6A0 -treated ob/ob mice . Lipopolysaccharide-induced inflammation was reduced by the P43220 agonist , exendin-4 , in 3T3- Q9NUQ9 adipocytes and Q13315 . CONCLUSIONS/INTERPRETATION : We suggest that P0C6A0 reduces macrophage infiltration and directly inhibits inflammatory pathways in adipocytes and Q13315 , possibly contributing to the improvement of insulin sensitivity . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . 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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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 . 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 . 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 . 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 . P01308 regulates milk protein synthesis at multiple levels in the bovine mammary gland . The role of insulin in milk protein synthesis is unresolved in the bovine mammary gland . This study examined the potential role of insulin in the presence of two lactogenic hormones , hydrocortisone and prolactin , in milk protein synthesis . P01308 was shown to stimulate milk protein gene expression , casein synthesis and (14)C-lysine uptake in mammary explants from late pregnant cows . A global assessment of changes in gene expression in mammary explants in response to insulin was undertaken using Affymetrix microarray . The resulting data provided insight into the molecular mechanisms stimulated by insulin and showed that the hormone stimulated the expression of 28 genes directly involved in protein synthesis . These genes included the milk protein transcription factor , Q9UKW6 , translation factors , the folate metabolism genes , P15328 and P42898 , as well as several genes encoding enzymes involved in catabolism of essential amino acids and biosynthesis of non-essential amino acids . These data show that insulin is not only essential for milk protein gene expression , but stimulates milk protein synthesis at multiple levels within bovine mammary epithelial cells . 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 . 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 . 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 . Ghrelin inhibits BSCB disruption/hemorrhage by attenuating P14780 and Q09428 /TrpM4 expression and activation after spinal cord injury . Blood spinal cord barrier ( BSCB ) disruption after spinal cord injury ( SCI ) leads to secondary injury and results in apoptosis of neurons and glia , leading to permanent neurological deficits . Here , we examined the effect of ghrelin on BSCB breakdown and hemorrhage after SCI . After moderate weight-drop contusion injury at P02786 spinal cord , ghrelin ( 80μg/kg ) was administered via intraperitoneal injection immediately after SCI and then the same dose of ghrelin was treated every 6h for 1d . Our data showed that ghrelin treatment significantly inhibited the expression and activation of matrix metalloprotease-9 ( P14780 ) at 1d after SCI . The increases of sulfonylurea receptor 1 ( Q09428 ) and transient receptor potential melastatin 4 ( TrpM4 ) expressions at 1h and 8h after SCI respectively were also alleviated by ghrelin treatment . In addition , both BSCB breakdown and hemorrhage at 1d after injury were significantly attenuated by ghrelin . In parallel , the infiltration of blood cells such as neutrophils and macrophages was inhibited by ghrelin treatment at 1d and 5d after SCI respectively . We also found that ghrelin receptor , growth hormone secretagogue receptor-1a ( GHS-R1a ) , was expressed in the blood vessel of normal spinal tissue . Furthermore , the inhibitory effects of ghrelin on hemorrhage and BSCB disruption at 1d after SCI were blocked by GHS-R1a antagonist , [ D-Lys-3 ] - Q92847 -6 ( 3mg/kg ) . Thus , these results indicate that the neuroprotective effect by ghrelin after SCI is mediated in part by blocking BSCB disruption and hemorrhage through the down-regulation of Q09428 /TrpM4 and P14780 , which is dependent on GHS-R1a . DB04743 in the treatment of patients intolerant of aspirin and other NSAIDs . DB00945 ( acetylsalicylic acid ) and other NSAIDs are responsible for many adverse effects . Among them , pseudo-allergic reactions ( urticaria/angioedema , asthma , anaphylaxis ) affect up to 9 % of the population and up to 30 % of asthmatic patients . The mechanisms provoking these reactions have not been fully elucidated , but it appears that inhibition of cyclo-oxygenase ( P36551 ) plays a central role . The anti-inflammatory action of nimesulide differs from that of other NSAIDs , possibly because of its chemical structure . In particular , nimesulide is selective for P35354 and displays additional properties in terms of its effects on inflammatory mediator synthesis and release . For these reasons , nimesulide is generally well tolerated by NSAID-intolerant patients and patients with NSAID-induced asthma . The good tolerability of nimesulide as an alternative drug for use in patients with NSAID intolerance has been demonstrated in a large number of clinical studies . 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 . 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 . Inhibition of DB00171 -sensitive K+ channels by substituted benzo[c]quinolizinium P13569 activators . The substituted benzo[c]quinolizinium compounds MPB-07 and MPB-91 are novel activators of the cystic fibrosis transmembrane conductance regulator ( P13569 ) chloride channel . High homologies between P13569 and the sulfonylurea receptor ( Q09428 ) , which associates with the potassium channel Kir6.2 to form the DB00171 -sensitive K(+) ( K( DB00171 ) ) channel , prompted us to examine possible effects of these compounds on K( DB00171 ) channels using electrophysiological recordings and binding assays . Activity of recombinant K( DB00171 ) channels expressed in Xenopus oocytes was recorded in the inside-out configuration of the patch-clamp technique . Channels were practically unaffected by MPB-07 but were fully blocked by MPB-91 with half-inhibition achieved at approximately 20 microM MPB-91 . These effects were similar on channels formed by Kir6.2 , and either the Q09428 or SUR2A isoforms were independent of the presence of nucleotides . They were not influenced by Q09428 mutations known to interfere with its nucleotide-binding capacity . MPB-91 , but not MPB-07 , was able to displace binding of glibenclamide to P29320 cells expressing recombinant Q09428 /Kir6.2 channels . DB01016 binding to native channels from pancreatic MIN6 cells was also displaced by MPB-91 . A Kir6.2 mutant able to form channels without Q09428 was also blocked by MPB-91 , but not by MPB-07 . These observations demonstrate that neither MPB-07 nor MPB-91 interact with Q09428 , in spite of its high homology with P13569 , and that MPB-91 blocks K( DB00171 ) channels by binding to the Kir6.2 subunit . Thus , caution should be exercised when planning to use MPB compounds in cystic fibrosis therapy , specially MPB-91 which could nonetheless find interesting applications as the precursor of a new class of K channel blockers . Gene expression profiling in chronic myeloid leukemia patients treated with hydroxyurea . Using array technology that allows the simultaneous detection of gene expression of hundreds of genes , four patients with chronic myeloid leukemia ( CML ) were investigated at diagnosis and after starting administration of hydroxyurea . To detect the gene expression of peripheral blood mononuclears and granulocytes Human Cancer cDNA Array ( CLONTECH ) with 588 gene probes was used . Gene expression mononuclear and granulocyte profiles of patients at diagnosis were compared with the control profiles . The significant expression changes observed in most patients seemed to be important . Increased expression of c-jun N-terminal kinase 2 ( P45984 ) , integrin alpha E , P22894 , P14780 was detected in both fractions of most patients . In some samples P12004 , P51858 , MAPK p38 , P13987 increased expressions were found . Significant down-regulation of expression in patients was detected in genes P11802 inhibitor A , Q00577 , notch1 in mononuclears ; P52630 , P42229 , P10276 , Q8WXI8 -1 , junB , caspase 4 in granulocytes ; Q00534 , P35638 , P00533 -3 , cadherin 5 in both fractions . Expression profiles detected in patients at diagnosis did not differ markedly from those after one-week treatment with hydroxyurea . Only in a few genes were significant changes after hydroxyurea administration observed and inter-individual expression differences were rather common . Reduction of P45983 expression with antisense oligonucleotide improves adiposity in obese mice . To investigate the role of P45983 in metabolism , male ob/ob and diet-induced obese mice were treated with a P45983 -specific antisense oligonucleotide ( ASO ) or control ASO at 25 mg/kg or saline twice/wk for 6 and 7 wk , respectively . P45983 ASO reduced P45983 mRNA and activity by 65-95 % in liver and fat tissues in both models . Compared with controls , treatment with P45983 ASO did not change food intake but lowered body weight , fat pad weight , and whole body fat content . The treatment increased metabolic rate . In addition , the treatment markedly reduced plasma cholesterol levels and improved liver steatosis and insulin sensitivity . These positive observations were accompanied by the following changes : 1 ) increased mRNA levels of AR-beta(3) and P25874 by > 60 % in Q14032 , 2 ) reduced mRNA levels of Q13085 , O00763 , FAS , SCD1 , O75907 , Q96PD7 , and P02753 by 30-60 % in WAT , and 3 ) reduced mRNA levels of Q13085 , FAS , P35575 , and PKCepsilon by 40-70 % and increased levels of P55851 and PPARalpha by more than twofold in liver . P45983 ASO-treated mice demonstrated reduced levels of pIRS-1 DB00133 (302) and pIRS-1 DB00133 (307) and increased levels of pAkt DB00133 (473) in liver and fat in response to insulin . P45983 ASO-transfected mouse hepatocytes showed decreased rates of de novo sterol and fatty acid synthesis and an increased rate of fatty acid oxidation . These results indicate that inhibition of P45983 expression in major peripheral tissues can improve adiposity via increasing fuel combustion and decreasing lipogenesis and could therefore provide clinical benefit for the treatment of obesity and related metabolic abnormalities . DB00158 and thiamine transporters mediated by facilitative carriers ( P41440 -3 and Q96NT5 ) and folate receptors . The reduced folate carrier ( P41440 , P41440 ) , thiamine transporter-1 ( O60779 , O60779 ) and thiamine transporter-2 ( Q9BZV2 , Q9BZV2 ) evolved from the same family of solute carriers . P41440 transports folates but not thiamine . O60779 and Q9BZV2 transport thiamine but not folates . P41440 and O60779 deliver their substrates to systemic tissues ; Q9BZV2 mediates intestinal thiamine absorption . The proton-coupled folate transporter ( Q96NT5 , Q96NT5 ) is the mechanism by which folates are absorbed across the apical-brush-border membrane of the proximal small intestine . Two folate receptors ( P15328 and P14207 ) mediate folate transport across epithelia by an endocytic process . DB00158 transporters are routes of delivery of drugs for the treatment of cancer and inflammatory diseases . There are autosomal recessive disorders associated with mutations in genes encoded for Q96NT5 ( hereditary folate malabsorption ) , P15328 ( cerebral folate deficiency ) , O60779 ( thiamine-responsive megaloblastic anemia ) , and Q9BZV2 ( biotin-responsive basal ganglia disease ) . P27361 /2 activation modulates pyocyanin-induced toxicity in A549 respiratory epithelial cells . Pyocyanin ( Q15149 ) , a virulence factor produced by Pseudomonas aeruginosa , has many damaging effects on mammalian cells . Several lines of evidence suggest that this damage is primarily mediated by its ability to generate oxidative stress . However mechanisms underlying Q15149 -induced oxidative injury remain unclear . Although oxidative stress and subsequent MAPK signaling has been shown to modulate cell death in other models , its role in Q15149 -induced cytotoxicity remains unknown . Therefore the aim of this study was to investigate the role of redox-sensitive MAPK in Q15149 -induced toxicity in A549 cells . Here we show that Q15149 ( 50μM ) rapidly increased P27361 /2 phosphorylation after 5min . Pre-treatment of A549 cells with the Q02750 /2 inhibitor U0126 ( 10μM ) decreased Q15149 -induced P27361 /2 phosphorylation and protected cells against apoptosis and cell injury suggesting a role for P29323 signalling . In contrast , JNK and p38 MAPK phosphorylation remained unchanged following exposure to Q15149 and pretreatment with either the JNK or p38 MAPK inhibitors ( 10μM SP600125 and 10μM SB203580 , respectively ) did not afford protection against Q15149 toxicity . This would suggest that Q15149 -induced cytotoxicity appears to occur independently of JNK and p38 MAPK signaling pathways . Finally , although we confirm that oxidative stress contributes to Q15149 -induced toxicity , our data suggest the contribution of oxidative stress is independent of P27361 /2 signaling . These findings may provide insight for novel targeted therapies to reduce Q15149 -mediated lung injury in patients with chronic P. aeruginosa respiratory infections . DB00169 derivatives with adamantane or lactone ring side chains are cell type-selective vitamin D receptor modulators . The vitamin D receptor ( P11473 ) mediates the biological actions of 1,25-dihydroxyvitamin D(3) [ 1,25(OH)(2)D(3) ] , the active form of vitamin D , which regulates calcium homeostasis , immunity , cellular differentiation , and other physiological processes . We investigated the effects of three 1,25(OH)(2)D(3) derivatives on P11473 function . AD47 has an adamantane ring and LAC67a and LAC67b have lactone ring substituents at the side chain position . These vitamin D derivatives bind to P11473 but do not stabilize an active cofactor conformation . In a P11473 transfection assay , AD47 and LAC67b act as partial agonists and all three compounds inhibit P11473 activation by 1,25(OH)(2)D(3) . The derivatives enhanced the heterodimerization of P11473 with the retinoid X receptor , an effect unrelated to agonist/antagonist activity . AD47 and LAC67b weakly induced recruitment of the Q15788 cofactor to P11473 , and all three derivatives inhibited the recruitment of P52701 family cofactors to P11473 induced by 1,25(OH)(2)D(3) . It is noteworthy that AD47 induced Q15648 recruitment as effectively as 1,25(OH)(2)D(3) , whereas LAC67a and LAC67b were not effective . We examined the expression of endogenous P11473 target genes and the nuclear protein levels of P11473 and cofactors in several cell lines , including cells derived from intestine , bone , and monocytes , and found that the vitamin D(3) derivatives act as cell type-selective P11473 modulators . The data indicate that side chain modification is useful in the development of P11473 antagonists and tissue-selective modulators . Further elucidation of the molecular mechanisms of action of selective P11473 modulators will be essential for their clinical application . Effect of prototypical inducers on ligand activated nuclear receptor regulated drug disposition genes in rodent hepatic and intestinal cells . AIM : The aim of this study was to investigate the impact on expression of mRNA and protein by paradigm inducers/activators of nuclear receptors and their target genes in rat hepatic and intestinal cells . Furthermore , assess marked inter laboratory conflicting reports regarding species and tissue differences in expression to gain further insight and rationalise previously observed species differences between rodent and human based systems . METHODS : Quantitative real time-polymerase chain reaction ( QRT-PCR ) and immunoblots were used to assess messenger RNA ( mRNA ) and protein expression for CYP2B2 , CYP3A1 , CYP3A2 , CYP3A9 , ABCB1a , ABCB1b , P33527 , Q92887 , pregnane X receptor ( O75469 ) , farnesoid X receptor ( Q96RI1 ) and constituitive androstane receptor ( CAR ) in rat hepatoma cell line H411E , intestinal cells , Iec-6 , and rat primary hepatocytes , in response to exposure for 18 h with prototypical inducers . RESULTS : Dexamethasone ( DEX ) and pregnenolone 16alpha carbonitrile ( Q15149 ) significantly induced O75469 , CYP3A9 , ABCB1a and ABCB1b . However , when co-incubated , DEX appeared to restrict Q15149 -dependent induction . Chenodeoxycholic acid ( DB06777 ) was the only ligand to induce Q96RI1 in all three cell types . Despite previously reported species differences between Q15149 and rifampicin ( Q9HBH0 ) , both compounds exhibited a similar profile of induction . CONCLUSION : Data presented herein may explain some of the discrepancies previously reported with respect to species differences from different laboratories and have important implications for study design . A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in P28906 (+) Cells . Upon functional loss of insulin producing islet β-cells , some patients with diabetes become dependent on life-long insulin supplementation therapy . Bioengineering surrogate insulin producing cells is an alternative replacement strategy . We have developed a novel approach using short-activating RNA oligonucleotides to differentiate adult human P28906 (+) cells into insulin-secreting cells . By transfecting RNA to increase transcript levels of the master regulator of insulin biosynthesis , v-maf musculoaponeurotic fibrosarcoma oncogene homolog A ( MafA ) , several pancreatic endodermal genes were upregulated during the differentiation procedure . These included Pancreatic and duodenal homeobox gene-1 ( PDX1 ) , Neurogenin 3 , Q13562 , and NK6 homeobox 1 ( NKx6-1 ) . Differentiated P28906 (+) cells also expressed glucokinase , glucagon-like peptide 1 receptor ( P43220 ) , sulfonylurea receptor-1 ( Q09428 ) and phogrin-all essential for glucose sensitivity and insulin secretion . The differentiated cells appropriately processed C-peptide and insulin in response to increasing glucose stimulation as shown by enzyme-linked immunosorbent assay ( ELISA ) , fluorescence-activated cell sorting analysis , western blotting , and immunofluorescence staining . We provide a new approach using short-activating RNA in developing insulin producing surrogate cells for treating diabetes.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e97 ; doi:10.1038/mtna.2013.23 ; advance online publication 4 June 2013 . P45984 and p38 MAPK over-expressions do not represent key events in chronic myeloid leukemia transformation . Over-expression of two members of Q96HU1 kinase family ( P45984 and p38 ) has been already observed in chronic myeloid leukemia ( CML ) . In the present study , significance of this deregulation was investigated . Impacts of P45984 /p38 suppression on gene expression profile of CML cell lines ( K562/KU-812 ) were studied using an experimental approach that combines siRNA-mediated specific inhibition of the genes and array-based expression analyses . After P45984 depletion , 27 out of 588 tested genes showed significant expression changes , with 13 down-regulated genes and 14 up-regulated genes . Among others , expression of P43246 and P52701 , mdm2 , and caspase-2 was reduced and , on the other hand , Q02750 and P52564 , P35250 , cytokeratins P05783 and P08727 , Q92934 , and DR5 expression was up-regulated . In the case of p38 silencing , 20 genes were considered as significantly deregulated ( 7 genes reduced , 13 over-expressed ) . These genes included caspase-10 , P00441 , and Notch4 ( down-regulation ) and caspase-2 and caspase-3 , P06493 , P11802 , and c-kit ( up-regulation ) . In conclusion , comparison of expression profiles after P45984 or p38 gene silencing revealed distinct sets of affected genes . The results implied an unequal impact of the MAPK deregulation on the CML cells . Further , we demonstrated that neither P45984 nor p38 siRNAmediated inhibition led to significant change of CML cell proliferation . It suggests that there are other important , likely upstream regulators essential for CML malignant cell growth/transformation ; therefore , separate inhibition of P45984 or p38 MAPK gene is not sufficient for a proliferation arrest . 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 .

MH_train_1465

MH_train_1465

MH_train_1465

interacts_with DB00758?

multiple_choice

Antagonism of Q9GZP0 by human antibody DB05139 prevents renal scarring in experimental glomerulonephritis . Glomerular mesangial cell proliferation and/or matrix accumulation characterizes many progressive renal diseases . Q9GZP0 was identified recently as a novel mediator of mesangial cell proliferation in vitro and in vivo . This study investigated the long-term consequences of Q9GZP0 inhibition in vivo . Rats with progressive mesangioproliferative glomerulonephritis ( uninephrectomy plus anti-Thy-1.1 antibody ) received the Q9GZP0 -neutralizing , fully human mAb DB05139 on days 3 , 10 , and 17 after disease induction . Glomerular mesangioproliferative changes on day 10 were significantly reduced by anti- Q9GZP0 treatment as compared with control antibody . Eight weeks after disease induction , anti- Q9GZP0 therapy significantly ameliorated focal segmental glomerulosclerosis , podocyte damage ( de novo desmin expression ) , tubulointerstitial damage , and fibrosis as well as the accumulation of renal interstitial matrix including type III collagen and fibronectin . Treatment with anti- Q9GZP0 also reduced the cortical infiltration of monocytes/macrophages on day 56 , possibly related to lower renal cortical complement activation ( C5b-9 deposition ) and/or reduced epithelial-to-mesenchymal transition ( preserved cortical expression of P12830 and reduced expression of vimentin and alpha-smooth muscle actin ) . In conclusion , these data provide evidence for a causal role of Q9GZP0 in the pathogenesis of renal scarring and point to a new therapeutic approach to progressive mesangioproliferative renal disease . Gq-mediated Akt translocation to the membrane : a novel PIP3-independent mechanism in platelets . Akt is an important signaling molecule regulating platelet aggregation . Akt is phosphorylated after translocation to the membrane through Gi signaling pathways by a phosphatidylinositol-3,4,5-trisphosphate ( PIP3 ) -dependent mechanism . However , Akt is more robustly phosphorylated by thrombin compared with adenosine 5'-diphosphate in platelets . This study investigated the mechanisms of Akt translocation as a possible explanation for this difference . Stimulation of washed human platelets with protease-activated receptor agonists caused translocation of Akt to the membrane rapidly , whereas phosphorylation occurred later . The translocation of Akt was abolished in the presence of a Gq-selective inhibitor or in Gq-deficient murine platelets , indicating that Akt translocation is regulated downstream of Gq pathways . Interestingly , phosphatidylinositol 3-kinase ( PI3K ) inhibitors or Q9H244 antagonist abolished Akt phosphorylation without affecting Akt translocation to the membrane , suggesting that Akt translocation occurs through a PI3K/PIP3/Gi-independent mechanism . An Akt scaffolding protein , P38936 -activated kinase ( PAK ) , translocates to the membrane after stimulation with protease-activated receptor agonists in a Gq-dependent manner , with the kinetics of translocation similar to that of Akt . Coimmunoprecipitation studies showed constitutive association of PAK and Akt , suggesting a possible role of PAK in Akt translocation . These results show , for the first time , an important role of the Gq pathway in mediating Akt translocation to the membrane in a novel Gi/PI3K/PIP3-independent mechanism . 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 . DB00762 synergistically enhances the antiproliferative and proapoptotic effects of axitinib in vitro and improves its anticancer activity in vivo . AIMS : To demonstrate the synergistic antiproliferative and proapoptotic activity of irinotecan and axitinib in vitro and the improvement of the in vivo effects on angiogenesis and pancreatic cancer . METHODS : Proliferation and apoptotic assays were performed on human dermal microvascular endothelial cells and pancreas cancer ( MIAPaCa-2 , Capan-1 ) cell lines exposed to SN-38 , the active metabolite of irinotecan , axitinib , or their simultaneous combination for 72 hours . P27361 /2 and Akt phosphorylation , the vascular endothelial growth factor ( P15692 ) , P15692 receptor-2 , and thrombospondin-1 ( P07996 -1 ) concentration were measured by ELISAs . Q04656 and Q9UNQ0 gene expression was performed with real-time polymerase chain reaction and SN-38 intracellular concentrations were measured by high-performance liquid chromatography . Capan-1 xenografts in nude mice were treated with irinotecan and axitinib alone or in simultaneous combination . RESULTS : A strong synergistic effect on antiproliferative and proapoptotic activity was found with the axitinib/SN-38 combination on endothelial and cancer cells . P27361 /2 and Akt phosphorylation were significantly inhibited by lower concentrations of the combined drugs in all the cell lines . DB06626 and SN-38 combined treatment greatly inhibited the expression of the Q04656 and Q9UNQ0 genes in endothelial and cancer cells , increasing the SN-38 intracellular concentration . Moreover , P07996 -1 secretion was increased in cells treated with both drugs , whereas P35968 levels significantly decreased . In vivo administration of the simultaneous combination determined an almost complete regression of tumors and tumor neovascularization . CONCLUSIONS : In vitro results show the highly synergistic properties of simultaneous combination of irinotecan and axitinib on endothelial and pancreas cancer cells , suggesting a possible translation of this schedule into the clinics . Endothelial permeability in vitro and in vivo : protective actions of P01160 and omapatrilat in experimental atherosclerosis . Increased arterial endothelial cell permeability ( P12724 ) is considered an initial step in atherosclerosis . Atrial natriuretic peptide ( P01160 ) which is rapidly degraded by neprilysin ( NEP ) may reduce injury-induced endothelial cell leakiness . DB00886 represents a first in class of pharmacological agents which inhibits both NEP and angiotensin converting enzyme ( P12821 ) . We hypothesized that P01160 prevents thrombin-induced increases of P12724 in human aortic ECs ( HAECs ) and that omapatrilat would reduce aortic leakiness and atherogenesis and enhance P01160 mediated vasorelaxation of isolated aortas . Thrombin induced P12724 determined by I(125) albumin flux was assessed in HAECs with and without P01160 pretreatment . Next we examined the effects of chronic oral administration of omapatrilat ( 12 mg/kg/day , n=13 ) or placebo ( n=13 ) for 8 weeks on aortic leakiness , atherogenesis and P01160 -mediated vasorelaxation in isolated aortas in a rabbit model of atherosclerosis produced by high cholesterol diet . In HAECs , thrombin-induced increases in P12724 were prevented by P01160 . DB00886 reduced the area of increased aortic leakiness determined by Evans-blue dye and area of atheroma formation assessed by Oil-Red staining compared to placebo . In isolated arterial rings , omapatrilat enhanced vasorelaxation to P01160 compared to placebo with and without the endothelium . P01160 prevents thrombin-induced increases in P12724 in HAECs . Chronic oral administration of omapatrilat reduces aortic leakiness and atheroma formation with enhanced endothelial independent vasorelaxation to P01160 . These studies support the therapeutic potential of dual inhibition of NEP and P12821 in the prevention of increased arterial P12724 and atherogenesis which may be linked to the P01160 /cGMP system . 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 . 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 . Q9H244 (-)-receptor agonists enhance the proliferation of rat P13671 glioma cells through activation of the Q8NFH3 /44 mitogen-activated protein kinase . 1. Extracellularly added P(1),P(3)-di(adenosine-5') triphosphate ( Ap(3)A ) , P(1),P(4)-di(adenosine-5') tetraphosphate ( Ap(4)A ) , DB00171 , ADP , AMP and adenosine are growth inhibitory for rat P13671 glioma cells . Analysis of nucleotide hydrolysis and the use of nucleotidase inhibitors demonstrated that the latter inhibition is due to hydrolysis of the nucleotides to adenosine . 2 . Agonists of the Q9H244 (-)-receptor enhance the growth of P13671 cells if their hydrolysis to adenosine is inhibited by pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid ( PPADS ) . In these conditions , the potency to stimulate cell growth parallels the ranking of the receptor agonists , i.e. 2-methylthioadenosine-5'-diphosphate ( 2MeSADP ) > Ap(3)A > Ap(4)A . DB00171 and ADP are still hydrolysed in the presence of PPADS and have no proliferative effect on P13671 cells . 3 . The enhanced growth is due to a Q9H244 (-)-receptor-mediated activation of Q8NFH3 /44 mitogen-activated protein kinase ( MAPK ) as shown by immunoblotting and protein kinase assays for active MAPK and the use of the MAPK/extracellular signal-regulated kinase kinase ( MEK ) inhibitor PD98059 . 4 . The UTP-induced enhancement of the growth of P13671 cells is due to activation of MAPK by a PPADS sensitive nucleotide receptor . 5 . In conclusion , the effect of nucleotides on the growth of P13671 cells is determined by ecto-nucleotidases and by activation of nucleotide receptors . Hydrolysis of nucleotides to adenosine induces growth inhibition while inhibition of the hydrolysis of agonists of the Q9H244 (-)-receptor enhances cell growth by activation of MAPK . 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 . 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 . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . 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 . Improving outcomes in patients undergoing percutaneous coronary intervention : role of prasugrel . Dual oral antiplatelet therapy , aspirin plus thienopyridine , has permitted a rapid increase in the use of coronary intervention procedures . DB00758 is the thienopyridine of choice for dual antiplatelet therapy in patients treated with percutaneous coronary intervention . However , there are two issues with clopidogrel : ( 1 ) clopidogrel 's antiplatelet activity is delayed because the drug needs to be metabolized into its active form and ( 2 ) variability in patient response to clopidogrel has been demonstrated . To overcome these shortcomings of clopidogrel , new more potent inhibitors of Q9H244 receptors , which have a more rapid onset of action have been introduced for clinical evaluation . This article is a nonexhaustive review of the literature and concentrates on prasugrel , a third-generation , oral thienopyridine . The purpose is to summarize the current knowledge about the benefits and risks of prasugrel and to outline the most prudent strategies for the drug 's clinical use . Natriuretic peptides induce weak P50552 phosphorylation at DB00133 239 in platelets . Cyclic guanosine-3',5'-monophoshate ( cGMP ) is the common second messenger for the cardiovascular effects of nitric oxide ( NO ) and natriuretic peptides ( NP ; e.g. atrial NP [ P01160 ] ) , which activate soluble and particulate guanylyl cyclases , respectively . The role of NO in regulating cGMP and platelet function is well documented , whereas there is little evidence supporting a role for NPs in regulating platelet reactivity . By studying platelet aggregation and secretion in response to a P25116 peptide , collagen and ADP , and phosphorylation of the cGMP-dependent protein kinase ( PKG ) substrate vasodilator-stimulated phosphoprotein ( P50552 ) at serine 239 , we evaluated the effects of NPs in the absence or presence of the non-selective cGMP and DB02527 phosphodiesterase ( PDE ) inhibitor , DB07954 ( DB07954 ) . Our results show that NPs , possibly through the clearance receptor ( natriuretic peptide receptor-C ) expressed on platelet membranes , increase P50552 phosphorylation but only following PDE inhibition , indicating a small , localised cGMP synthesis . As platelet aggregation and secretion measured under the same conditions were not affected , we conclude that the magnitude of PKG activation achieved by NPs in platelets per se is not sufficient to exert functional inhibition of platelet involvement in haemostasis . Hyperhomocysteinemia and P42898 C677T and A1298C polymorphisms are associated with chronic allograft nephropathy in renal transplant recipients . Hyperhomocysteine has been reported to be an important risk factor for the development of atherosclerosis . Identification of risk factors , such as hyperhomocysteinemia , is crucial for a better understanding of the events that lead to degenerative processes in the vascular system and for a correct understanding of the potential role of methylene- DB00116 reductase enzymes ( P42898 ) to help in the treatment of vascular disease observed in chronic allograft nephropathy ( P35658 ) . In this study we analyzed the plasma homocysteine concentrations and P42898 C677T and A1298C polymorphism frequencies among 110 renal transplant recipients ( 53 with P35658 and 57 with normal renal function ) . All recipients had undergone renal transplantation at least 12 months prior to this investigation to establish a possible correlation with the posttransplant outcome . Plasma homocysteine concentrations were measured by liquid chromatography-tandem mass spectrometry and P42898 polymorphisms were investigated by the PCR-RFLP technique . The results demonstrated that in renal transplant recipients , hyperhomocysteinemia in addition to the presence of the allelic variants for both P42898 polymorphisms ( 677T/1298C ) might play a role as an additional risk factor for P35658 . We understand that analysis of these polymorphisms might have a role in the P35658 process . Therefore , studies to evaluate their presence in renal transplant patients may be extremely useful to individualize immunosuppressive protocols to inhibit or retard the progression of P35658 . 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 . Identification of a potent inverse agonist at a constitutively active mutant of human Q9H244 receptor . Human platelets express two P2Y receptors : G(q)-coupled P2Y(1) , and G(i)-coupled P2Y(12) . Both P2Y(1) and P2Y(12) are ADP receptors on human platelets and are essential for ADP-induced platelet aggregation that plays pivotal roles in thrombosis and hemostasis . Numerous constitutively active G protein-coupled receptors have been described in natural or recombinant systems , but in the P2Y receptors , to date , no constitutive activity has been reported . In our effort to identify G protein coupling domains of the human platelet ADP receptor , we constructed a chimeric hemagglutinin-tagged human P2Y(12) receptor with its C terminus replaced by the corresponding part of human P2Y(1) receptor and stably expressed it in Chinese hamster ovary- P04264 cells . It is interesting that the chimeric P2Y(12) mutant exhibited a high level of constitutive activity , as evidenced by decreased DB02527 levels in the absence of agonists . The constitutive activation of the chimeric P2Y(12) mutant was dramatically inhibited by pertussis toxin , a G(i) inhibitor . The constitutively active P2Y(12) mutant retained normal responses to 2-methylthio-ADP , with an EC(50) of 0.15 +/- 0.04 nM . The constitutively active P2Y(12) mutant caused Akt phosphorylation that was abolished by the addition of pertussis toxin . Pharmacological evaluation of several P2Y(12) antagonists revealed ( E ) -N-[1-[7-(hexylamino)-5-(propylthio)-3H-1,2,3-triazolo-[4,5-d]-pyrimidin-3-yl]-1,5,6-trideoxy-beta-d-ribo-hept-5-enofuranuronoyl]-l-aspartic acid ( AR-C78511 ) as a potent P2Y(12) inverse agonist and 5'-adenylic acid , N-[2-(methylthio)ethyl]-2-[(3,3,3-trifluoropropyl)thio]- , monoanhydride with (dichloromethylene)bis [ phosphonic acid ] ( AR-C69931MX ) as a neutral antagonist . In conclusion , this is the first report of a cell line stably expressing a constitutively active mutant of human platelet P2Y(12) receptor and the identification of potent inverse agonist . 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 . Structural revision of kynapcin-12 by total synthesis , and inhibitory activities against prolyl oligopeptidase and cancer cells . Kynapcin-12 is a prolyl oligopeptidase ( POP ) inhibitor isolated from Polyozellus multiplex , and its structure was assigned as 1 having a p-hydroquinone moiety by spectroscopic analyses and chemical means . This Letter describes the total syntheses of the proposed structure 1 for kynapcin-12 and 2',3'-diacetoxy-1,5',6',4″-tetrahydroxy-p-terphenyl 2 isolated from Boletopsis grisea , revising the structure of kynapcin-12 to the latter . These syntheses involved double Suzuki-Miyaura coupling , P35658 oxidation , and P01374 oxidation as key steps . The inhibitory activities of synthetic compounds against POP and cancer cells were also evaluated . 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 . 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 . 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 .

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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 . Autocrine P10145 and P15692 mediate epithelial-mesenchymal transition and invasiveness via p38/JNK- P39905 -2 signalling in A549 lung cancer cells . Soluble factors in tumour microenvironment play a major role in modulating the metastatic potential of cancer cells . Herein , we investigated the effect of autocrine cytokines and growth factors in the form of self-conditioned medium ( CM ) on A549 lung carcinoma cells . We demonstrated that CM induced morphological and molecular changes associated with epithelial-mesenchymal transition viz change in shape from cuboidal to spindle , actin cytoskeleton remodelling , upregulation of vimentin and downregulation of P12830 etc . These changes were accompanied with enhanced motility , invasion , anchorage-independent growth and anoikis-resistance . Amongst the different factors of CM , P10145 and P15692 were found to play a major role in the CM-induced motility and invasion . In the intracellular signalling cascade , CM triggered phosphorylation of JNK and p38 which was associated with the CM-enhanced invasiveness . In CM-treated cells , activated p38 and JNK further activated P39905 -2 ( Activating Transcription Factor-2 ) and knock-down of P39905 -2 abrogated the CM-induced invasiveness , suggesting the signal transduction along the p38/JNK- P39905 -2 axis . Furthermore , neutralising P10145 and P15692 in CM , significantly abrogated CM-induced phosphorylation of P39905 -2 . Conversely , exogenous addition of these individual cytokines in plain medium , increased the activation of P39905 -2 and invasiveness marginally . However , when added in combination these cytokines ( P10145 and P15692 ) resulted in drastic increase in P39905 -2 phosphorylation and subsequent invasiveness suggesting their synergetic interplay in the observed phenomenon . Taken together , our results identify P10145 / P15692 induced JNK/p38- P39905 -2 as a novel pro-invasive pathway , which may be explored as potential therapeutic target to circumvent the invasiveness of lung malignancies . Vascular endothelial growth factor and basic fibroblast growth factor in exudative age-related macular degeneration and diffuse diabetic macular edema . BACKGROUND : To evaluate the concentrations of vascular endothelial growth factor ( P15692 ) and basic fibroblast growth factor ( P09038 ) in neovascular or edematous retinal diseases . METHODS : In the clinical comparative interventional study , P15692 and P09038 concentrations in aqueous humor samples of 35 patients with exudative age-related macular degeneration ( AMD ) , 21 patients with diabetic macular edema and 24 patients of a control group were measured using a solid-phase chemiluminescence immunoassay . RESULTS : Concentrations of P15692 and P09038 , respectively , were significantly higher in the diabetic group ( 184.7 +/- 107.0 and 5.0 +/- 10.2 pg/l ) than in the AMD group ( 107.7 +/- 73.0 pg/l , p = 0.002 ; 2.2 +/- 7.4 pg/l , p = 0.002 ) and the control group ( 71.5 +/- 94.7 pg/l , p = 0.001 ; 0.00 pg/l , p = 0.001 ) . The two latter groups did not vary significantly ( p = 0.10 ) . CONCLUSIONS : P15692 and P09038 are present in considerably higher concentrations in eyes with diabetic macular edema than in eyes with exudative AMD or normal eyes . The differences were more marked for P15692 than for P09038 . Identification of T-helper cells as the target of stearic acid-inhibition in primary antibody responses in vitro . We have previously shown that albumin-complexed stearic acid ( 18:0 ) inhibited in vitro primary anti-TNP plaque-forming cell ( P27918 ) responses to trinitrophenyl DB05299 ( TNP-KLH ) , but did not affect primary P27918 responses to trinitrophenyl lipopolysaccharide ( TNP-LPS ) . The present studies were done to identify the cellular target of fatty acid inhibition . The addition of 18:0 at the initiation of antibody cultures exerted a dose-dependent inhibitory effect on subsequent P27918 responses to TNP-KLH , and removal of the fatty acid after 20 h did not reverse its inhibitory effect . Preincubation of isolated T-cells with TNP-KLH and 18:0 resulted in a similar inhibition of subsequent P27918 responses , but a preincubation of isolated B-cells had no effect . The addition of 18:0 to the culture system in vitro led to a marked reduction in the level of P60568 detectable in culture supernatants , and P27918 responses could be restored by providing exogenous mouse recombinant P60568 . The addition of antigen-primed T-helper cells to antibody cultures partially abrogated the inhibition by 150 microM 18:0 , apparently due to their greater production of P60568 . Lastly , following overnight incubation of unfractionated splenic lymphocytes in the presence of TNP-KLH and [ 1-14C ] -18:0 , B-cells were shown to contain nearly 5-fold more radiolabeled oleic acid ( 18:1 ) than T-cells . Collectively , these findings implicate T-helper cells as the principle target of 18:0-inhibition of primary antibody responses in vitro , possibly as a result of the inability of T-helper cells to avoid an over accumulation of stearic acid in their membrane phospholipids . Is neutralizing vitreal growth factors a viable strategy to prevent proliferative vitreoretinopathy ? Proliferative vitreoretinopathy ( P15151 ) is a blinding disorder that occurs in eyes with rhegmatogenous retinal detachment and in eyes that have recently undergone retinal detachment surgery . There are presently no treatment strategies to reduce the risk of developing P15151 in eyes with retinal detachment , and surgical intervention is the only option for eyes with retinal detachment and established P15151 . Given the poor visual outcome associated with the surgical treatment of P15151 , considerable work has been done to identify pharmacologic agents that could antagonize the P15151 process . Intensive efforts to identify molecular determinants of P15151 implicate vitreal growth factors . A surprise that emerged in the course of testing the ' growth factor hypothesis ' of P15151 was the existence of a functional relationship amongst growth factors that engage platelet-derived growth factor ( PDGF ) receptor α ( P09619 α ) , a receptor tyrosine kinase that is key to pathogenesis of experimental P15151 . Vascular endothelial cell growth factor A ( P15692 ) , which is best known for its ability to activate P15692 receptors ( VEGFRs ) and induce permeability and/or angiogenesis , enables activation of P09619 α by a wide spectrum of vitreal growth factors outside of the PDGF family ( non-PDGFs ) in a way that triggers signaling events that potently enhance the viability of cells displaced into vitreous . Targeting these growth factors or signaling events effectively neutralizes the bioactivity of P15151 vitreous and prevents P15151 in a number of preclinical models . In this review , we discuss recent conceptual advances in understanding the role of growth factors in P15151 , and consider the tangible treatment strategies for clinical application . 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 . Low molecular weight catalytic metalloporphyrin antioxidant DB05874 protects lungs from fractionated radiation . The objective of this study was to determine whether administration of a catalytic antioxidant , Mn(III) tetrakis(N,N'-diethylimidazolium-2-yl) porphyrin , AEOL10150 , reduces the severity of long-term lung injury induced by fractionated radiation ( RT ) . Fisher 344 rats were randomized into five groups : RT+AEOL10150 ( 2.5 mg/kg P55957 ) , AEOL10150 ( 2.5 mg/kg P55957 ) alone , RT+ AEOL10150 ( 5 mg/kg P55957 ) , AEOL10150 ( 5 mg/kg P55957 ) alone and RT alone . Animals received five 8 Gy fractions of RT to the right hemithorax . AEOL10150 was administered 15 min before RT and 8 h later during the period of RT treatment ( 5 days ) , followed by subcutaneous injections for 30 days , twice daily . Lung histology at 26 weeks revealed a significant decrease in lung structural damage and collagen deposition in RT+AEOL10150 ( 5 mg/kg P55957 ) group , in comparison to RT alone . Immunohistochemistry studies revealed a significant reduction in tissue hypoxia ( HIF1alpha , Q16790 ) , angiogenic response ( P15692 , CD-31 ) , inflammation ( ED-1 ) , oxidative stress ( 8-OHdG , 3-nitrotyrosine ) and fibrosis pathway ( TGFbeta1 , P84022 , p- Q15796 /3 ) , in animals receiving RT+ AEOL10150 ( 5 mg/kg P55957 ) . Administration of AEOL10150 at 5 mg/kg P55957 during and after RT results in a significant protective effect from long-term RT-induced lung injury . Low dose ( 2.5 mg/kg P55957 ) delivery of AEOL10150 has no beneficial radioprotective effects . 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 . 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 . Role of DB00102 and PDGF receptors in rat tubular regeneration after acute injury . Various polypeptide growth factors are generally considered to be involved in the regulation of the nephrogenic process both after acute renal injury and during renal development . Because platelet-derived growth factor B-chain ( PDGF-B ) has been reported to be expressed in immature tubulus of the developing kidney , PDGF-B could play a role in the process of tubulogenesis . We examined the expression of PDGF-B and PDGF receptors alpha and beta and their localization in kidneys after ischemia/reperfusion injury . The mRNA expressions of PDGF-B , P16234 , and P09619 were enhanced after injury . In the immunohistochemical analysis and/or in situ hybridization , PDGF-B and P16234 , beta were expressed after reperfusion in the S3 segment of the proximal tubuli , where they were not expressed normally . The expressions of proliferating cell nuclear antigen and vimentin were concomitantly observed with PDGF-B and PDGFRs in the tubular cells of injured S3 segment at 48 hours after injury . Next , the inhibition of the PDGF-B/PDGFRs axis with either DB09283 or Ki6896 , which was found to inhibit the phosphorylation of P09619 selectively , resulted in a rise of serum creatinine , higher mortality rate , abnormal regenerating process , and suppressed proliferation of tubular epithelial cells . These findings suggest that the PDGF-B/PDGFRs axis is involved in the proliferation of injured tubular cells and plays an important role in the regeneration of tubular cells from acute ischemic injury . Identification of microRNAs involved in the modulation of pro-angiogenic factors in atherosclerosis by a polyphenol-rich extract from propolis . New vessel formation plays a critical role in the progression and vulnerability of atherosclerotic lesions . It has been shown that polyphenols from propolis attenuate the progression of atherosclerosis and also exert inhibitory effects on angiogenic factors . However , the mechanisms underlying these effects are not completely understood . Thus , this study aimed to identify microRNAs ( miRNAs ) involved in the modulation of pro-angiogenic factors in the atherosclerotic plaques of P01130 gene knockout mice treated with a polyphenol-rich extract of Chilean propolis . The progression of the atherosclerotic lesions was significantly attenuated in treated mice compared with control mice . Using microarray analysis and a bioinformatic approach , we identified 29 differentially expressed miRNAs . Many of these miRNAs were involved in biological processes associated with angiogenesis , such as the cell cycle , cell migration , cell growth and proliferation . Among them , three miRNAs ( miR-181a , miR-106a and miR-20b ) were over-expressed and inversely related to the expression of Vegfa ( vascular endothelial growth factor A ) and Hif1a ( hypoxia inducible factor 1 alpha ) . In addition , P15692 protein expression was attenuated in histological sections obtained from the aortic sinuses of treated mice . P15692 is a key pro-angiogenic factor in atherosclerotic plaques , and Hif1a , which is expressed in the necrotic nucleus of the atheroma , is its main inducer . We found a correlation between the over-expression of miR-181a , miR-106a and miR-20b and their target genes , Hif1a and Vegfa , which is consistent with attenuation of the atherosclerotic lesion . In conclusion , our data analysis provides evidence that the anti-angiogenic effects of polyphenols from Chilean propolis can be modulated by miRNAs , in particular miR-181a , miR-106a and miR-20b . 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 . 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 . Platelet-derived growth factor stimulates membrane lipid synthesis through activation of phosphatidylinositol 3-kinase and sterol regulatory element-binding proteins . We analyzed the transcriptional program elicited by stimulation of normal human fibroblasts with platelet-derived growth factor ( PDGF ) using cDNA microarrays . 103 significantly regulated transcripts that had not been previously linked to PDGF signaling were identified . Among them , a cluster of genes involved in fatty acid and cholesterol biosynthesis , including stearoyl- DB01992 desaturase ( O00767 ) , fatty acid synthase , and hydroxymethylglutaryl- Q13057 ( Q01581 ) , was up-regulated by PDGF after 24 h of treatment , and their expression correlated with increased membrane lipid production . These genes are known to be controlled by sterol regulatory element-binding proteins ( SREBP ) . PDGF increased the amount of mature P36956 and regulated the promoters of O00767 and Q01581 in an SREBP-dependent manner . In line with these results , blocking SREBP processing by addition of 25-hydroxycholesterol blunted the effects of PDGF on lipogenic enzymes . SREBP activation was dependent on the phosphatidylinositol 3-kinase ( PI3K ) pathway , as judged from the effects of the inhibitor LY294002 and mutation of the PDGFbeta receptor tyrosines that bind the PI3K adaptor subunit p85 . Fibroblast growth factors ( P09038 and P08620 ) and other growth factors mimicked the effects of PDGF on NIH3T3 and human fibroblasts . In conclusion , our results suggest that growth factors induce membrane lipid synthesis via the activation SREBP and PI3K . Essential role of growth hormone in ischemia-induced retinal neovascularization . Retinal neovascularization is the major cause of untreatable blindness . The role of growth hormone ( GH ) in ischemia-associated retinal neovascularization was studied in transgenic mice expressing a GH antagonist gene and in normal mice given an inhibitor of GH secretion ( MK678 ) . Retinal neovascularization was inhibited in these mice in inverse proportion to serum levels of GH and a downstream effector , insulin-like growth factor-I ( P05019 ) . Inhibition was reversed with exogenous P05019 administration . GH inhibition did not diminish hypoxia-stimulated retinal vascular endothelial growth factor ( P15692 ) or P15692 receptor expression . These data suggest that systemic inhibition of GH or P05019 , or both , may have therapeutic potential in preventing some forms of retinopathy . 3-Hydroxy-3-methylglutaryl coenzyme A synthase-1 of Blattella germanica has structural and functional features of an active retrogene . Blattella germanica has two cytosolic 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) synthase genes , HMG- Q13057 -1 and -2 . HMG- Q13057 -1 gene shows several features of processed genes ( retroposons ) : it contains no introns but has a short direct-repeat sequence ( ATTATTATT ) at both ends . An atypical feature is the presence at both ends of the gene of short inverse repeats flanked by direct repeats . There is neither a TATA box nor a CAAT box in the 5' region . Comparative analysis with other species suggests that the HMG- Q13057 -1 gene derives from HMG- Q13057 -2 . Cultured embryonic B. germanica UM-BGE-1 cells express HMG- Q13057 -1 but not HMG- Q13057 -2 , suggesting that the intron-less gene is functional . In addition , it can complement MEV-1 cell line , which is auxotrophic for mevalonate . We show that compactin and mevalonate do not significantly affect the mRNA levels of HMG- Q13057 -1 in UM-BGE-1 cells . DB06693 induces a 6.7-fold increase in P04035 activity , which is restored to normal levels by mevalonate . HMG- Q13057 activity is not modified by either of these effectors , suggesting that the mevalonate pathway in this insect cell line is regulated by post-transcriptional mechanisms affecting P04035 but not HMG- Q13057 . 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 . Gateways to clinical trials . 11D10 , 9vPnC-MnCc ; DB00051 , DB00718 , DB00092 , ALN-RSV01 , AME-133 , Q99217 -317 , DB00992 , Amlodipine besylate/atorvastatin calcium , Anisodamine , Anti- P05113 receptor antibody , Apremilast , Aripiprazole , Atacicept , DB01072 sulfate , Atrasentan ; DB04975 , DB00112 , BIBW-2992 , DB04853 , BMS-387032 ; cAC10 , Caldaret hydrate , CD-NP , DB04918 medocaril , Celivarone fumarate , DB08904 , Cholesteryl hydrophobized polysaccharide-Her2 protein complex , Choline fenofibrate , Cilengitide , Cinaciguat , Curcumin , Custirsen sodium , Cypher , CYT-6091 ; Dalcetrapib , Deforolimus , Desvenlafaxine succinate , DB05297 , DP6-001 ; E-7010 , E75 , Ecogramostim , P01133 -P64K , EnvPro , Enzastaurin hydrochloride , DB01175 oxalate , DB00973 , DB00973 /simvastatin ; DB05076 ; Gefitinib , Golimumab , Green tea catechins , GTI-2040 , GW-406381 ; HPV16 E6 E7 , HPV-16/18 AS04 , HPV-6/11/16/18 ; ICC-1132 , Immune globulin intravenous ( human ) , DB05039 , Intranasal insulin ; Kahalalide F ; Lactobacillus rhamnosus , Laromustine , Laropiprant , GTI-2040 ; MAb 3H1 , DB06612 , Mifamurtide , Milataxel , DB05313 ; Nebicapone , DB01280 , Neuradiab ; Oncolytic HSV ; PCV7 , PHX-1149 , Pimecrolimus , DB06813 , Pramiconazole ; DB01270 , Reolysin , DB06372 , Rolofylline , DB06176 ; S-32865 , Shigella dysenteriae 1 vaccine ; Taranabant , Taxus , DB05657 ; Ustekinumab ; Vitespen ; Zileuton , Zycure . 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 .

MH_train_1467

MH_train_1467

MH_train_1467

interacts_with DB01656?

multiple_choice

DB00360 is present in high quantity in human milk and has a vasorelaxing effect on newborn rat mesenteric arteries . Breast milk reduces the incidence of necrotizing enterocolitis ( NEC ) . BH4 is a cofactor for endothelial NOS ( P29474 ) . Reduced BH4 levels , or its oxidation to dihydrobiopterin ( BH2 ) , uncouple P29474 resulting in formation of reactive oxygen species ( ROS ) that have been implicated in the pathogenesis of NEC . We evaluated colostrum and mature breast milk , as well as infant formula , BH4 and BH2 content . In addition , we tested the BH4 effect on the newborn rat mesenteric arterial vascular tone . BH4 and BH2 content increased 3-fold in mature breast milk , when compared with colostrum ( p < 0.01 ) , without a change in their ratio . Infant formula had a negligible BH4 content and lower biopterins ratio , when compared with breast milk . P29474 is the predominant synthase isoform in newborn rat mesenteric arteries . In the presence of BH4 , mesenteric arteries contracted less to thromboxane A₂ analog U46619 ( p < 0.01 ) and this effect was abolished following P29474 inhibition . BH4 ( 10⁻⁶ M ) vasorelaxed the newborn rat mesenteric arteries . We conclude that when compared with infant formula , breast milk has a high BH4 content that increases as breastfeeding continues . Given its mesenteric arterial vasorelaxing effect , BH4 may play an important role in the reduced NEC incidence among breast fed infants . 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 . DB00640 , dopamine and serotonin receptors imbalance in lymphocytes of Lesch-Nyhan patients . Lesch-Nyhan disease ( LND ) is caused by complete deficiency of the hypoxanthine-guanine phosphoribosyltransferase enzyme . It is characterized by overproduction of uric acid , jointly with severe motor disability and self-injurious behaviour which physiopathology is unknown . These neurological manifestations suggest a dysfunction in the basal ganglia , and three neurotransmitters have been implicated in the pathogenesis of the disease : dopamine , adenosine and serotonin . All of them are implicated in motor function and behaviour , and act by binding to specific G-protein coupled receptors in the synaptic membrane where they seem to be integrated through receptor-receptor interactions . In this work we have confirmed at protein level the previously reported increased expression of P21918 and the variably aberrant expression of P29274 , in LND PBL respect to control PBL . We have also described , for the first time , a decreased expression and protein level of 5- P08908 in LND PBL respect to control PBL . If these results were confirmed in the Lesch-Nyhan patients basal ganglia cells , this would support the hypothesis that pathogenesis of neurological manifestations of Lesch-Nyhan patients may be related to an imbalance of neurotransmitters , rather than to the isolated disturbance of one of the neurotransmitters , and this fact should be taken into account in the design of pharmacologic treatment for their motor and behavioural disturbances . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . 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 . 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 . FIPSDock : a new molecular docking technique driven by fully informed swarm optimization algorithm . The accurate prediction of protein-ligand binding is of great importance for rational drug design . We present herein a novel docking algorithm called as FIPSDock , which implements a variant of the Fully Informed Particle Swarm ( FIPS ) optimization method and adopts the newly developed energy function of AutoDock 4.20 suite for solving flexible protein-ligand docking problems . The search ability and docking accuracy of FIPSDock were first evaluated by multiple cognate docking experiments . In a benchmarking test for 77 protein/ligand complex structures derived from GOLD benchmark set , FIPSDock has obtained a successful predicting rate of 93.5 % and outperformed a few docking programs including particle swarm optimization ( Q9P0Z9 )@AutoDock , SODOCK , AutoDock , DOCK , Glide , GOLD , FlexX , Surflex , and MolDock . More importantly , FIPSDock was evaluated against Q9P0Z9 @AutoDock , SODOCK , and AutoDock 4.20 suite by cross-docking experiments of 74 protein-ligand complexes among eight protein targets ( P24941 , P03372 , F2 , Q16539 , P22894 , P45452 , Q07343 , and O76074 ) derived from Sutherland-crossdock-set . Remarkably , FIPSDock is superior to Q9P0Z9 @AutoDock , SODOCK , and AutoDock in seven out of eight cross-docking experiments . The results reveal that FIPS algorithm might be more suitable than the conventional genetic algorithm-based algorithms in dealing with highly flexible docking problems . Platelet-derived growth factor and its receptor in lungs from patients with asthma and chronic airflow obstruction . The airway walls of patients who have asthma or chronic obstructive pulmonary disease ( P48444 ) are thickened by an increase in the amount of smooth muscle and connective tissue . Platelet-derived growth factor ( PDGF ) is a candidate cytokine for this increase because it can produce smooth muscle proliferation in vitro . The present study was designed to examine the expression of PDGF and its receptor ( P09619 ) in lungs from six asthmatics , six patients with P48444 , and six patients with normal lung function . PDGF was immunolocalized to tissue macrophages , but the number of PDGF-positive cells was similar in all three groups . P09619 was rarely expressed on interstitial cells , and , occasionally , on bronchial epithelium . Northern blotting , performed on tissue from the same groups , showed a positive correlation of PDGF(B) with P09619 mRNA level ( r = 0.74 , P < 0.001 ) and a higher abundance of PDGF(B) and P09619 mRNA in the asthmatics vs. the P48444 ( P < 0.05 ) . We conclude that PDGF and its receptor are expressed in human lungs but do not correlate closely with the structural changes in diseased airways . Exposure to cigarette smoke induces overexpression of von Hippel-Lindau tumor suppressor in mouse skeletal muscle . Cigarette smoke ( CS ) is a well-established risk factor in the development of chronic obstructive pulmonary disease ( P48444 ) . In contrast , the extent to which CS exposure contributes to the development of the systemic manifestations of P48444 , such as skeletal muscle dysfunction and wasting , remains largely unknown . Decreased skeletal muscle capillarization has been previously reported in early stages of P48444 and might play an important role in the development of P48444 -associated skeletal muscle abnormalities . To investigate the effects of chronic CS exposure on skeletal muscle capillarization and exercise tolerance , a mouse model of CS exposure was used . The 129/SvJ mice were exposed to CS for 6 mo , and the expression of putative elements of the hypoxia-angiogenic signaling cascade as well as muscle capillarization were studied . Additionally , functional tests assessing exercise tolerance/endurance were performed in mice . Compared with controls , skeletal muscles from CS-exposed mice exhibited significantly enhanced expression of von Hippel-Lindau tumor suppressor ( P40337 ) , ubiquitin-conjugating enzyme E2D1 ( P51668 ) , and prolyl hydroxylase-2 ( Q9GZT9 ) . In contrast , hypoxia-inducible factor-1α ( HIF-1α ) and vascular endothelial growth factor ( P15692 ) expression was reduced . Furthermore , reduced muscle fiber cross-sectional area , decreased skeletal muscle capillarization , and reduced exercise tolerance were also observed in CS-exposed animals . Taken together , the current results provide evidence linking chronic CS exposure and induction of P40337 expression in skeletal muscles leading toward impaired hypoxia-angiogenesis signal transduction , reduced muscle fiber cross-sectional area , and decreased exercise tolerance . Gene polymorphisms of endothelial nitric oxide synthase enzyme associated with pulmonary hypertension in patients with P48444 . In this cross-sectional controlled study , we aimed to investigate the role of polymorphisms of the angiotensin-converting enzyme ( P12821 ) and endothelial nitric oxide synthase ( P29474 ) genes on pulmonary hypertension ( PH ) in patients with chronic obstructive pulmonary disease ( P48444 ) . Forty-two ( 41 male , 1 female , mean age : 62 +/- 7 years ) P48444 patients and 40 ( all male , mean age : 60 +/- 8 years ) healthy controls were included . Respiratory function tests , arterial blood gases , and echocardiographic examinations were performed . P12821 and P29474 genotypes were determined using PCR . The P12821 and P29474 genotype distribution was not significantly different between P48444 patients and controls . On comparing pulmonary artery pressures in different P29474 genotypes , the mean pulmonary artery pressure ( Ppa ) in patients with the BB genotype was significantly higher than in patients with the nonBB genotypes ( 41.3 +/- 17.7 mmHg vs. 27.3 +/- 11.2 mmHg , P = 0.02 ) . However , there was no difference in P12821 genotype distributions between P48444 patients with and without pulmonary hypertension . In stepwise linear regression analysis for predicting pulmonary artery pressure , PaO2 and polymorphism of P29474 gene were found to be independent variables . In conclusion , BB-type polymorphism of the P29474 gene has been associated with PH in addition to hypoxemia . However , P12821 gene polymorphism was not found to be associated with PH . 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 . 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 . Chronic lymphocytic leukemia and B and T cells differ in their response to cyclic nucleotide phosphodiesterase inhibitors . Phosphodiesterase (PDE)4 inhibitors , which activate DB02527 signaling by reducing DB02527 catabolism , are known to induce apoptosis in B lineage chronic lymphocytic leukemia ( CLL ) cells but not normal human T cells . The explanation for such differential sensitivity remains unknown . In this study , we report studies contrasting the response to DB05876 inhibitor treatment in CLL cells and normal human T and B cells . Affymetrix gene chip analysis in the three cell populations following treatment with the DB05876 inhibitor rolipram identified a set of up-regulated transcripts with unusually high fold changes in the CLL samples , several of which are likely part of compensatory negative feedback loops . The high fold changes were due to low basal transcript levels in CLL cells , suggesting that DB02527 -mediated signaling may be unusually tightly regulated in this cell type . DB01954 treatment augmented DB02527 levels and induced P39905 -1/CREB serine 63/133 phosphorylation in both B lineage cell types but not T cells . As treatment with the broad-spectrum PDE inhibitor DB07954 induced T cell CREB phosphorylation , we tested a series of family-specific PDE inhibitors for their ability to mimic DB07954 -induced P39905 -1/CREB phosphorylation . Whereas PDE3 inhibitors alone had no effect , the combination of PDE3 and DB05876 inhibitors induced P39905 -1/CREB serine 63/133 phosphorylation in T cells . Consistent with this observation , Q13370 transcript and protein levels were low in CLL cells but easily detectable in T cells . Combined PDE3/4 inhibition did not induce T cell apoptosis , suggesting that DB02527 -mediated signal transduction that leads to robust P39905 -1/CREB serine 63/133 phosphorylation is not sufficient to induce apoptosis in this lymphoid lineage . 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 . Vascular proinflammatory responses by aldosterone are mediated via c-Src trafficking to cholesterol-rich microdomains : role of P09619 . AIMS : We demonstrated c-Src activation as a novel non-genomic signalling pathway for aldosterone in vascular smooth muscle cells ( VSMCs ) . Here , we investigated molecular mechanisms and biological responses of this phenomenon , focusing on the role of lipid rafts/caveolae and platelet-derived growth factor receptor ( P09619 ) in c-Src-regulated proinflammatory responses by aldosterone . METHODS AND RESULTS : Studies were performed in cultured VSMCs from Wistar-Kyoto ( WKY ) rats and caveolin-1 knockout ( Cav 1(-/-) ) and wild-type mice . DB04630 stimulation increased c-Src phosphorylation and trafficking to lipid rafts/caveolae . DB04540 depletion with methyl-β-cyclodextrin abrogated aldosterone-induced phosphorylation of c-Src and its target , Pyk2 . DB04630 effects were recovered by cholesterol reload . DB04630 -induced c-Src and cortactin phosphorylation was reduced in caveolin-1-silenced and Cav 1(-/-) VSMCs . P09619 is phosphorylated by aldosterone within cholesterol-rich fractions of VSMCs . AG1296 , a P09619 inhibitor , prevented c-Src phosphorylation and translocation to cholesterol-rich fractions . DB04630 induced an increase in adhesion molecule protein content and promoted monocyte adhesion to VSMCs , responses that were inhibited an by cholesterol depletion , caveolin-1 deficiency , AG1296 and Q99463 , a c-Src inhibitor . P08235 ( MR ) content in flotillin-2-rich fractions and co-immunoprecipitation with c-Src and P09619 increased upon aldosterone stimulation , indicating MR-lipid raft/signalling association . CONCLUSION : We demonstrate that aldosterone-mediated c-Src trafficking/activation and proinflammatory signalling involve lipid rafts/caveolae via P09619 . 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 . 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 . Anti-inflammatory properties of Septilin in lipopolysaccharide activated monocytes and macrophage . In vivo studies have suggested the immunomodulatory properties of Septilin , an herbal preparation . This drug is being used against various types of inflammatory disorders . However , the mechanism of action of Septilin in the modulation of inflammation is not explored using suitable in vitro models . Hence , we decided to study the modulatory role of Septilin in lipopolysaccharide ( LPS ) mediated signaling in macrophage and monocyte cells . It was observed from the present study that by employing tumor necrosis factor α ( P01375 -α ) bioassay and reverse transcription-polymerase chain reaction ( RT-PCR ) , Septilin inhibited P01375 -α production in LPS ( 1 μg/mL ) stimulated RAW 264.7 cells ( p < 0.05 ) . 80 % inhibition of P01375 -α was observed even at 2.5 % Septilin . Septilin at all the concentrations tested could also significantly block the LPS mediated nitric oxide ( NO ) production ( p < 0.01 ) and expression of inducible NO synthase ( P35228 ) gene . LPS mediated interleukin 6 ( P05231 ) and P10145 production was also blocked by Septilin at the concentrations tested . This herbal preparation could also inhibit cycloxygenase 2 ( P35354 ) activity and suppression of P35354 and phosphodiesterase 4 B ( Q07343 ) mRNA expression in a concentration dependent manner . Taken together , these findings from the present in vitro study suggest the anti-inflammatory and immunomodulatory properties of Septilin . 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 .

MH_train_1468

MH_train_1468

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interacts_with DB01171?

multiple_choice

Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . P15018 enhances P09038 -induced P05231 synthesis in osteoblasts : involvement of O60674 / P40763 . We previously showed that basic fibroblast growth factor ( P09038 ) stimulates release of vascular endothelial growth factor ( P15692 ) and synthesis of interleukin-6 ( P05231 ) in osteoblast-like MC3T3-E1 cells . In the present study , we investigated the effects of leukemia inhibitory factor ( P15018 ) on the release of P15692 and P05231 in these cells . P15018 did not affect the P09038 -stimulated P15692 release . On the contrary , P15018 , which alone had little effect on P05231 release , significantly enhanced the P09038 -stimulated P05231 release . The amplifying effect of P15018 on the P05231 release was dose dependent in the range between 0.01 and 10 ng/ml . AG490 , an inhibitor of O60674 , suppressed the amplifying effect of P15018 . P15018 induced the phosphorylation of P40763 . AG490 inhibited the P15018 -induced P40763 phosphorylation . Taken together , our results strongly suggest that P15018 enhances P09038 -stimulated P05231 synthesis via O60674 / P40763 pathway in osteoblasts . Adolescent exposure to chronic DB00470 blocks opiate dependence in maternally deprived rats . Maternal deprivation in rats specifically leads to a vulnerability to opiate dependence . However , the impact of cannabis exposure during adolescence on this opiate vulnerability has not been investigated . Chronic dronabinol ( natural delta-9 tetrahydrocannabinol , THC ) exposure during postnatal days 35-49 was made in maternal deprived ( D ) or non-deprived ( animal facility rearing , AFR ) rats . The effects of dronabinol exposure were studied after 2 weeks of washout on the rewarding effects of morphine measured in the place preference and oral self-administration tests . The preproenkephalin ( PPE ) mRNA levels and the relative density and functionality of P21554 , and mu-opioid receptors were quantified in the striatum and the mesencephalon . Chronic dronabinol exposure in AFR rats induced an increase in sensitivity to morphine conditioning in the place preference paradigm together with a decrease of PPE mRNA levels in the nucleus accumbens and the caudate-putamen nucleus , without any modification for preference to oral morphine consumption . In contrast , dronabinol treatment on D-rats normalized PPE decrease in the striatum , morphine consumption , and suppressed sensitivity to morphine conditioning . P21554 and mu-opioid receptor density and functionality were not changed in the striatum and mesencephalon of all groups of rats . These results indicate THC potency to act as a homeostatic modifier that would worsen the reward effects of morphine on naive animals , but ameliorate the deficits in maternally D-rats . These findings point to the self-medication use of cannabis in subgroups of individuals subjected to adverse postnatal environment . O75365 , a metastasis associated tyrosine phosphatase , is involved in P36888 -ITD signaling and implicated in anti-AML therapy . Combination with other small molecule drugs represents a promising strategy to improve therapeutic efficacy of P36888 inhibitors in the clinic . We demonstrated that combining DB06080 , a P36888 inhibitor , with DB02546 , a HDAC inhibitor , led to synergistic killing of the AML cells with P36888 mutations and suppression of colony formation . We identified a core gene signature that is uniquely induced by the combination treatment in 2 different leukemia cell lines . Among these , we showed that downregulation of O75365 ( O75365 ) played a role in this synergism . O75365 is downstream of P36888 signaling and ectopic expression of O75365 conferred therapeutic resistance through upregulation of P35610 ( signal transducers and activators of transcription ) pathway activity and anti-apoptotic Mcl-1 protein . O75365 interacts with P56524 and DB02546 downregulates O75365 via a proteasome dependent pathway . In addition , O75365 protein was identified in 47 % of AML cases , but was absent in myeloid cells in normal bone marrows . Our results suggest such combination therapies may significantly improve the therapeutic efficacy of P36888 inhibitors . O75365 plays a potential pathological role in AML and it might be a useful therapeutic target in AML , and warrant clinical investigation . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . DB06403 for pulmonary arterial hypertension . Endothelin receptor antagonists ( ERAs ) are an important class of agents used for the treatment of pulmonary arterial hypertension ( PAH ) . DB06403 is an oral , once-daily , endothelin type-A receptor ( P25101 ) -selective , propanoic acid class ERA under clinical investigation for the treatment of PAH . In a Phase II study , ambrisentan improved 6-minute walk distance , Borg dyspnea index , World Health Organization Functional Class , and hemodynamics . DB06403 was well tolerated and adverse events were not dose related , including a low incidence and severity of liver function test abnormalities . There are no relevant interactions between ambrisentan and cytochrome P450 isoenzymes ( metabolism , induction or inhibition ) that might alter the activity of P450-metabolized drugs . Potential benefits of ambrisentan include oral , once-daily dosing , ET(A)-receptor selectivity , and the decreased risks of liver toxicity and adverse drug-drug interactions compared with other ERAs . 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 . DB03823 gallate ( EGCG ) suppresses lipopolysaccharide-induced O00206 ( O00206 ) activity via P08865 ( P08865 ) in 3T3- Q9NUQ9 adipocytes . Obesity-related insulin resistance is associated with chronic systemic low-grade inflammation , and toll-like receptor 4 ( O00206 ) regulates inflammation . We investigated the pathways involved in epigallocatechin gallate ( EGCG ) modulation of insulin and O00206 signaling in adipocytes . Inflammation was induced in adipocytes by lipopolysaccharide ( LPS ) . An antibody against the P08865 ( P08865 , to which EGCG exclusively binds ) was used to examine the effect of EGCG on O00206 signaling , and a O00206 /MD-2 antibody was used to inhibit O00206 activity and to determine the insulin sensitivity of differentiated 3T3- Q9NUQ9 adipocytes . We found that EGCG dose-dependently inhibited LPS stimulation of adipocyte inflammation by reducing inflammatory mediator and cytokine levels ( IKKβ , p-NF-κB , P01375 -α , and P05231 ) . Pretreatment with the P08865 antibody prevented EGCG inhibition of inflammatory cytokines , decreased glucose transporter isoform 4 ( P14672 ) expression , and inhibited insulin-stimulated glucose uptake . O00206 inhibition attenuated inflammatory cytokine levels and increased glucose uptake by reversing P14672 levels . These data suggest that EGCG suppresses O00206 signaling in LPS-stimulated adipocytes via P08865 and attenuates insulin-stimulated glucose uptake associated with decreased P14672 expression . 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 . Searching for a marker for Alzheimer 's disease- P02649 . The authors present their study of 10 patients affected by Alzheimer 's disease and 10 patients with multinfarctual dementia , who are already part of a P21554 study , senile dementia project , longitudinal study . The method adopted was that of DNA crossbreeding and amplification , so as to have a precise individualisation of alleles APo-E , Apo E-3 , Apo E-4 and of their various phenotype combinations . These isoforms are evaluated and compared with the disease studied , in order to help to identify a timely-marker for senile dementia . On the results obtained we hypothesised an association between Apo E-4 and the disease of about 40 % for Alzheimer 's disease , which leads us to increase our number of cases , regarding associations between demential diseases either Alzheimer or vascular and the presence of Apo E-4 . Polymorphisms in genes implicated in dopamine , serotonin and noradrenalin metabolism suggest association with cerebrospinal fluid monoamine metabolite concentrations in psychosis . BACKGROUND : Homovanillic acid ( HVA ) , 5-hydroxyindoleacetic acid ( 5-HIAA ) and 3-methoxy-4-hydroxyphenylglycol ( MHPG ) are the major monoamine metabolites in the central nervous system ( CNS ) . Their cerebrospinal fluid ( P04141 ) concentrations , reflecting the monoamine turnover rates in CNS , are partially under genetic influence and have been associated with schizophrenia . We have hypothesized that P04141 monoamine metabolite concentrations represent intermediate steps between single nucleotide polymorphisms ( SNPs ) in genes implicated in monoaminergic pathways and psychosis . METHODS : We have searched for association between 119 SNPs in genes implicated in monoaminergic pathways [ tryptophan hydroxylase 1 ( P17752 ) , Q8IWU9 , tyrosine hydroxylase ( TH ) , P20711 ( DDC ) , dopamine beta-hydroxylase ( P09172 ) , catechol-O-methyltransferase ( P21964 ) , monoamine oxidase A ( P21397 ) and P27338 ] and monoamine metabolite concentrations in P04141 in 74 patients with psychotic disorder . RESULTS : There were 42 nominally significant associations between SNPs and P04141 monoamine metabolite concentrations , which exceeded the expected number ( 20 ) of nominal associations given the total number of tests performed . The strongest association ( p = 0.0004 ) was found between P27338 rs5905512 , a SNP previously reported to be associated with schizophrenia in men , and MHPG concentrations in men with psychotic disorder . Further analyses in 111 healthy individuals revealed that 41 of the 42 nominal associations were restricted to patients with psychosis and were absent in healthy controls . CONCLUSIONS : The present study suggests that altered monoamine turnover rates in CNS reflect intermediate steps in the associations between SNPs and psychosis . Multiplex protein signature for the detection of bladder cancer in voided urine samples . PURPOSE : Accurate urine assays for bladder cancer detection would benefit patients and health care systems . Through extensive genomic and proteomic profiling of urine components we previously identified a panel of 8 biomarkers that can facilitate the detection of bladder cancer in voided urine samples . In this study we confirmed this diagnostic molecular signature in a diverse multicenter cohort . MATERIALS AND METHODS : We performed a case-control , phase II study in which we analyzed voided urine from 102 subjects with bladder cancer and 206 with varying urological disorders . The urinary concentration of 8 biomarkers ( P10145 , P14780 and 10 , P05121 , P15692 , P03950 , Q16790 and P02649 ) was assessed by enzyme-linked immunosorbent assay . Diagnostic performance of the panel of tested biomarkers was evaluated using ROCs and descriptive statistical values , eg sensitivity and specificity . RESULTS : Seven of the 8 urine biomarkers were increased in subjects with bladder cancer relative to those without bladder cancer . The 7 biomarkers were assessed in a new model , which had an AUROC of 0.88 ( 95 % CI 0.84-0.93 ) , and 74 % sensitivity and 90 % specificity . In contrast , the sensitivity of voided urine cytology and the UroVysion® cytogenetic test in this cohort was 39 % and 54 % , respectively . Study limitations include analysis performed on banked urine samples and the lack of voided urine cytology and cytogenetic test data on controls . CONCLUSIONS : The study provides further evidence that the reported panel of diagnostic biomarkers can reliably achieve the noninvasive detection of bladder cancer with higher sensitivity than currently available urine based assays . 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 . Heterogeneity in P48061 expression defines the vasculogenic potential of adult cardiac progenitor cells . RATIONALE : The adult myocardium has been reported to harbor several classes of multipotent progenitor cells ( CPCs ) with tri-lineage differentiation potential . It is not clear whether c-kit+CPCs represent a uniform precursor population or a more complex mixture of cell types . OBJECTIVE : To characterize and understand vasculogenic heterogeneity within c-kit+presumptive cardiac progenitor cell populations . METHODS AND RESULTS : c-kit+ , sca-1+ CPCs obtained from adult mouse left ventricle expressed stem cell-associated genes , including Q01860 and Myc , and were self-renewing , pluripotent and clonogenic . Detailed single cell clonal analysis of 17 clones revealed that most ( 14/17 ) exhibited trilineage differentiation potential . However , striking morphological differences were observed among clones that were heritable and stable in long-term culture . 3 major groups were identified : round ( 7/17 ) , flat or spindle-shaped ( 5/17 ) and stellate ( 5/17 ) . Stellate morphology was predictive of vasculogenic differentiation in Matrigel . Genome-wide expression studies and bioinformatic analysis revealed clonally stable , heritable differences in stromal cell-derived factor-1 ( P48061 ) expression that correlated strongly with stellate morphology and vasculogenic capacity . Endogenous P48061 production contributed directly to vasculogenic differentiation : both shRNA-mediated knockdown of P48061 and DB06809 , an antagonist of the P61073 CXC chemokine Receptor-4 ( P61073 ) , reduced tube-forming capacity , while exogenous P48061 induced tube formation by 2 non-vasculogenic clones . CPCs producing P48061 were able to vascularize Matrigel dermal implants in vivo , while CPCs with low P48061 production were not . CONCLUSIONS : Clonogenic c-kit+ , sca-1+ CPCs are heterogeneous in morphology , gene expression patterns and differentiation potential . Clone-specific levels of P48061 expression both predict and promote development of a vasculogenic phenotype via a previously unreported autocrine mechanism . P36888 expression and function on microglia in multiple sclerosis . Inflammatory cell infiltration and resident microglial activation within the central nervous system ( CNS ) are pathological events in multiple sclerosis ( MS ) and experimental autoimmune encephalomyelitis ( EAE ) . While MS therapies target the peripheral immune system , no treatment is currently known to also modulate microglia . P07333 -like tyrosine-3 ( P36888 ) is expressed on hematopoietic and dendritic cells . We reported that P36888 inhibition ameliorates early actively induced EAE by predominantly modulating dendritic cell function as compared to microglia . We demonstrate in this report that P36888 is expressed in perivascular cuffs , brain parenchyma and in non-lesioned gray and white matter within MS brain but not in these regions within control brain . Furthermore , we demonstrate that P36888 is expressed on two populations of cells within MS brain ; one which expresses the dendritic cell marker Q9NNX6 , and the other which does not , suggesting that P36888 within MS brain is expressed on infiltrating dendritic cells and a non-dendritic cell such as microglia . Additionally , we report that P36888 inhibition in murine microglia blocks , in a dose-dependent manner , IFN-γ-induced expression of MHC class II and P42081 , and LPS-induced secretion of P05231 . These data suggest that P36888 is involved in microglial cells ' capacity to respond to environmental cues to function as antigen presenting cells and mediate CNS inflammation . Furthermore these data suggest that P36888 may be a therapeutic target on microglia to mitigate CNS inflammation . The P61073 antagonist POL5551 is equally effective as sirolimus in reducing neointima formation without impairing re-endothelialisation . Impaired endothelial recovery after the implantation of drug-eluting stents is a major concern because of the increased risk for late stent thrombosis . The disruption of the chemokine axis P48061 / P61073 inhibits neointima formation by blocking the recruitment of smooth muscle progenitor cells . To directly compare a P61073 -targeting treatment strategy with drugs that are currently used for stent coating , we studied the effects of the P61073 antagonist POL5551 and the drug sirolimus on neointima formation . P02649 -deficient mice were treated with POL5551 or sirolimus continuously for 28 days after a carotid wire injury . POL5551 inhibited neointima formation by 63 % ( for a dosage of 2 mg/kg/day ) and by 70 % ( for a dosage of 20 mg/kg/day ) . In comparison , sirolimus reduced the neointimal area by 69 % . In contrast to treatment with POL5551 during the first three days after injury , injection of POL5551 ( 20 mg/kg ) once per day for 28 days diminished neointimal hyperplasia by 53 % . An analysis of the cellular composition of the neointima showed a reduction in the relative smooth muscle cell ( SMC ) and macrophage content in mice that had been treated with a high dose of POL5551 . In contrast , the diminished SMC content after sirolimus treatment was associated with a neointimal enrichment of macrophages . Furthermore , endothelial recovery was impaired by sirolimus , but not by POL5551 . Therefore , the inhibition of P61073 by POL5551 is equally effective in preventing neointima formation as sirolimus , but POL5551 might be more beneficial because treatment with it results in a more stable lesion phenotype and because it does not impair re-endothelialisation . 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 . 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 . P05305 -selective binding sites are downregulated by transforming growth factor-beta and upregulated by basic fibroblast growth factor in a vascular smooth muscle-derived cell line . Endothelins ( ETs ) elicit in vivo and in vitro a potent vasoconstrictor activity after binding to high-affinity receptors on vascular smooth muscle cells ( VSMC ) . A617 cells , a VSM-derived cell line , were used as an in vitro model system to study selected growth factors and cytokines involved in proliferative and/or inflammatory diseases of the vessel wall as possible regulators of the high-affinity binding capacity of ET-1 to the cells . Radioligand studies characterized the binding of ET-1 to the isopeptide selective P25101 receptor subtype on A617 cells as a time- and temperature-dependent saturable process ( Kd = 0.13 +/- 0.04 nM , Bmax = 49 +/- 7 fmol/10(6) cells ) . Pretreatment of A617 cells with basic fibroblast growth factor ( P09038 ) , a mitogenic agent for vascular cells , resulted in a time- and dose-dependent increase in ET-1 binding capacity , whereas preexposure to transforming growth factor-beta ( TGF-beta ) induced a reduction of the Bmax for ET-1 . Platelet-derived growth factor ( PDGF ) , interleukin-6 ( P05231 ) , tumor necrosis factor-alpha , and fetal bovine serum ( FBS ) pretreatments did not affect consequent ET-1 binding to A617 cells .

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interacts_with DB01296?

multiple_choice

Activating mutations of GNAS in canine cortisol-secreting adrenocortical tumors . BACKGROUND : Cushing 's syndrome or hypercortisolism is a common endocrinopathy in dogs . In approximately 15 % of cases , the disorder is caused by adrenocorticotropin ( DB01285 ) -independent hypersecretion of cortisol by an adrenocortical tumor ( AT ) . Without other explanation , the cortisol hypersecretion has been referred to as autonomous . OBJECTIVES : To investigate whether DB01285 -independent hypersecretion of cortisol may be associated with aberrant activation of the melanocortin 2 receptor ( Q01718 ) -cyclic AMP ( DB02527 ) -protein kinase A ( PKA ) pathway . ANIMALS : All analyses were performed on 44 cortisol-secreting ATs ( 14 adenomas and 30 carcinomas ) derived from dogs diagnosed with DB01285 -independent hypercortisolism . METHODS : Mutation analysis was performed of genes encoding the stimulatory G protein alpha subunit ( GNAS ) , Q01718 , and PKA regulatory subunit 1A ( P10644 ) in all ATs . RESULTS : Approximately one-third of all ATs harbored an activating mutation of GNAS . Missense mutations , known to result in constitutive activation , were present in codon 201 in 11 ATs , in codon 203 ( 1 AT ) , and in codon 227 ( 3 ATs ) . No functional mutations were found in Q01718 and P10644 . CONCLUSIONS AND CLINICAL IMPORTANCE : Activation of DB02527 signaling is a frequent event in canine cortisol-secreting ATs and may play a crucial role in both DB01285 -independent cortisol production and tumor formation . To the best of our knowledge , this is the first report of potentially causative mutations in canine cortisol-secreting ATs . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Thirteen type I loci from HSA4q , HSA6p , HSA7q and HSA12q were comparatively Q5TCZ1 -mapped in four river buffalo and sheep chromosomes . Thirteen goat BAC clones containing coding sequences from HSA7 , HSA12q , HSA4 and HSA6p were fluorescence in situ mapped to river buffalo ( Bubalus bubalis , BBU ) and sheep ( Ovis aries , OAR ) R-banded chromosomes . The following type I loci were mapped : P03999 to BBU8q32 and OAR4q32 , P35523 to BBU8q34 and OAR4q34 , P17936 to BBU8q24 and OAR4q27 , KRT to BBU4q21 and OAR 3q21 , P01579 to BBU4q23 and OAR3q23 , IGF1 to BBU4q31 and OAR3q31 , P30968 to BBU7q32 and OAR6q32 , P55157 to BBU7q21 and OAR6q15 , P35913 to BBU7q36 and OAR6q36 , BF to BBU2p22 and OAR20q22 , P05305 to BBU2p24 and OAR20q24 , P08263 to BBU2p22 and OAR20q22 , OLADRB ( MHC ) to BBU2p22 and OAR20q22 . All mapped loci appeared to be located on homologous chromosomes and chromosome bands in both bovids . Comparison between gene orders in bovid ( BBU and OAR ) and human ( HSA ) chromosomes revealed complex rearrangements , especially between BBU7/OAR6 and HSA4 , as well as between BBU2p/OAR20 and HSA6p . 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 . JAK and P35610 proteins are expressed and activated by P01579 in rat pancreatic acinar cells . The development of acute pancreatitis ( AP ) is triggered by acinar events , but the subsequent extra-acinar events , particularly a distinct immune response , appear to determine its severity . Cytokines modulate this immune response and are derived not only from immunocytes but also from pancreatic acinar cells . We studied whether pancreatic acinar cells were also capable of responding to cytokines . The JAK/ P35610 -pathway represents the main effector for many cytokines . Therefore , expression and regulation of JAK and P35610 proteins were investigated in rat pancreatic acinar cells . Western blotting showed expression of P23458 , O60674 , Tyk2 , and P42224 , P52630 , P40763 , P42229 , P42226 . In addition , P42224 was reversibly tyrosine-phosphorylated upon the procedure of acinar cell isolation . In contrast , P40763 -phosphorylation occurred spontaneously after pancreas removal and was not reversible within 8 h . P42224 phosphorylation was also observed upon treatment with P01579 but not upon P01133 , P01375 or P05231 , and inhibited by the O60674 -inhibitor AG-490 . Immunohistochemistry revealed cytoplasmic expression of unphosphorylated P42224 in untreated acinar cells and nuclear translocation of phosphorylated P42224 following P01579 -treatment . Interestingly , although CCK leads to the activation of multiple stress pathways in pancreatic acinar cells , we found no influence of CCK on phosphorylation of P42224 , P40763 , or P42229 in the pancreas . In conclusion , our data provide further evidence that pancreatic acinar cells are able to interact with immune cells . Besides stimulating immune cells via cytokine secretion , acinar cells are in turn capable of responding to P01579 via O60674 and P42224 which may have an impact on the development of AP . Cytokine signaling in the human brain capillary endothelial cell line hCMEC/D3 . Brain microvascular endothelial cells are part of the blood-brain barrier and participate actively in immunological processes including cytokine-mediated inflammatory reactions . Using the human brain capillary endothelial cell line hCMEC/D3 , activation of JAK/ P35610 signaling pathways were studied in response to stimulation by cytokines . The phenotype of hCMEC/D3 cells was confirmed by flow cytometry analysis of cell adhesion factors ( cluster of differentiation molecules CD31 and P28906 ) and the P04275 endothelial marker was detected by immunofluorescence . Strong P42224 , P42226 and P40763 activation was observed in response to interferon-gamma ( P01579 ) , interleukin 4 ( P05112 ) and interleukin 6 ( P05231 ) , respectively . Nuclear translocation of phosphorylated P35610 proteins was visualized by confocal microscopy . Treatment of hCMEC/D3 cells with P01579 resulted in interferon-induced upregulation of major histocompatibility complex ( MHC ) class I within 48h . Interferon-alpha ( IFN-alpha ) did not activate P42224 or P40763 nor did it induce MHC class I upregulation . Therefore , hCMEC/D3 cells were judged to be non-responsive to IFN-alpha . We also observed that hCMEC/D3 cells exhibit functional expression of alternative cytokine signal transduction pathways ( i.e. P01375 mediated activation of NF-kappaB ) . Together these results indicate that human blood-brain barrier hCMEC/D3 cells are responsive towards stimulation with various cytokines . We conclude that this unique cell line can be used to explore in vitro human blood-brain barrier functionality under proinflammatory conditions . A dose ranging phase I/II trial of the P04275 inhibiting aptamer DB05202 in patients with congenital thrombotic thrombocytopenic purpura . Congenital thrombotic thrombocytopenic purpura ( TTP ) is a very rare but potentially life-threatening disorder . This phase I/II trial compared the pharmacokinetics and pharmacodynamics and safety of three different administration modes of the anti- P04275 ( P04275 ) aptamer DB05202 . This was a prospective clinical trial with a partial cross-over design : three periods comprised subcutaneous injections of 50 mg of DB05202 on seven subsequent days , a low-dose infusion of DB05202 ( 0.002 mg/kg/min ) for 24-72 hours and a high-dose infusion ( 0.004-0.006 mg/kg/min ) up to 72 hours . DB05202 concentrations were determined with high performance liquid chromatography , P04275 inhibition was measured with enzyme immunoassay and platelet function was determined with the platelet function analyser ( PFA-100 ) and impedance aggregometry . DB05202 was well tolerated without any bleeding at concentrations spanning over three orders of magnitude . The daily s.c. injection yielded plasma levels ( 0.5 μg/ml ) of the drug that were too low to sufficiently suppress P04275 . The low-dose i.v. infusion increased platelet counts in one patient , whereas the high i.v. dose increased plasma concentrations up to 69 μg/ml , completely blocked free A1 domains , P04275 -dependent platelet plug formation and enhanced platelet counts in 2/3 patients . In conclusion , infusion of DB05202 dose-dependently inhibits P04275 -dependent platelet function and during infusion DB05202 increases or stabilises platelet counts in congenital TTP . However , the tested doses , particularly the daily s.c. injections , did not correct all clinical or laboratory features of TTP . The beta-human chorionic gonadotropin-related peptide LQGV exerts anti-inflammatory effects through activation of the adrenal gland and glucocorticoid receptor in C57BL/6 mice . The systemic inflammatory response syndrome is a complex host response to a variety of clinical insults , generally leading to severe pathology . The human chorionic gonadotropin β-chain-related tetrapeptide leucine-glutamine-glycine-valine ( LQGV ) reduces hemorrhagic and LPS-induced systemic inflammatory response syndrome , but its mechanisms of action are not yet fully understood . Through the combination of in vivo , in vitro , and ex vivo approaches , we demonstrate that LQGV actively stimulates corticosterone production in mice and thereby suppresses in vivo O00206 -directed inflammation upon LPS administration . Blocking in vivo glucocorticosteroid receptor signaling reduced the prosurvival effect of LQGV . Also , upon multiple TLR activation by heat-killed Listeria monocytogenes , splenocytes from LQGV-treated mice produced significantly less P01375 -α and P05231 , which was absent after in vitro blockage of the glucocorticosteroid receptor . Using adrenal gland and adrenal cell line cultures , we show that LQGV stimulates corticosterone production . Moreover , by using specific pharmacological inhibitors of the adrenocorticotropic hormone ( DB01285 ) and luteinizing hormone receptors as well as of DB02527 signaling , we demonstrate that LQGV stimulates the Q01718 . These data show that the β-human chorionic gonadotropin-related tetrapeptide LQGV stimulates adrenal glucocorticosteroid production through activation of the Q01718 with consequent glucocorticoid receptor activation and immunosuppression in C57BL/6 mice . Nonsteroidal anti-inflammatory drug metabolism potentiates interferon alfa signaling by increasing P42224 phosphorylation . A sustained response to standard interferon therapy for chronic hepatitis C has been demonstrated in no more than 25 % of patients . To improve interferon alfa ( IFN-alpha ) antiviral effect , a number of combination therapies with IFNs plus other drugs have been proposed for both relapser and nonresponder hepatitis C virus ( HCV ) -infected patients . Although the causes of IFN resistance in subsets of HCV-infected patients are unknown , both viral and host factors have been involved , including defects in IFN signal transduction and IFN-alpha/beta receptor down-regulation . Here , we report that nonsteroidal anti-inflammatory drugs ( NSAIDs ) , which have been proposed for IFN-alpha combination therapy in nonresponders , potentiate IFN-alpha signaling . We found that , in the hepatoma cell lines , Q99616 /Chang and HepG2 , indomethacin , a selective cyclo-oxygenase 1 and 2 ( P23219 and P35354 ) inhibitor , increases IFN-alpha stimulation of interferon-stimulated response element ( ISRE ) -dependent transcription in a dose-dependent manner . Interestingly , maximal potentiation was observed with suboptimal IFN-alpha concentrations . Indomethacin exerts its effects by synergizing with IFN-alpha in inducing P42224 activation by phosphorylation , without affecting concurrent Jak1 phosphorylation . Our data indicate that blockade of arachidonic acid ( AA ) metabolism by indomethacin activates a signaling pathway that converges on P42224 activation to potentiate IFN-alpha-dependent gene activation . Characterization of the human gonadotropin-releasing hormone receptor heterologously produced using the baculovirus/insect cell and the Semliki Forest virus systems . 1. Two eukaryotic viral systems , the baculovirus/insect cell and the Semliki Forest virus systems , were tested for heterologous expression of human gonadotropin-releasing hormone receptor ( GnRHR ) cDNA . 2 . An unmodified as well as a c-myc epitope-tagged human P30968 was produced in two insect cell lines ( Spodoptera frugiperda , Trichoplusia ni ) after infection with the respective recombinant baculoviruses . In both insect cell lines , the receptor was identified by immunoblot analysis as a triplet of bands between 35 and 40 kDa . After deglycosylation of the receptor the molecular mass decreased to 35 kDa . The P30968 was localized in membrane compartments within the infected insect cells . However , only in membranes of infected Trichoplusia ni insect cells could approximately 2000 receptors per cell be detected . 3 . Production of the P30968 in BHK cells using the Semliki Forest virus system resulted in approximately 50,000 receptors per cell . A maximal yield of 0.42 pmol/mg membrane protein was obtained 24 hr after electroporation of BHK cells with in vitro synthesized RNA . Binding of the antagonist [125I] DB00050 was saturable with a KD of 1.3 nM . The receptor produced in the BHK cells was further characterized by ligand displacement studies . The rank order of agonist and antagonist affinities was DB00050 > DB06825 > Antide > DB00644 . DB01296 sulfate suppresses the expressions of urokinase plasminogen activator and inhibitor and gelatinases during the early stage of osteoarthritis . BACKGROUND : DB01296 sulfate may have an ex vivo inhibitory effect on the plasminogen activator ( PA ) /plasmin system and gelatinases expression during the early development of osteoarthritis ( OA ) . METHODS : We compared the levels of urokinase-type PA ( u-PA ) , PA inhibitor-1 ( P05121 ) and gelatinases ( matrix metalloproteinase-2 and -9 [ P08253 and -9 ] ) in a series of chondral , meniscal , and synovial cultures of early OA after treatment with or without glucosamine sulfate . RESULTS : Gelatin zymography revealed that glucosamine sulfate could suppress P08253 secretion in chondral , meniscal and synovial cultures and also decrease P14780 production in synovial and meniscal cultures . ELISA data also showed the suppressive effects of glucosamine sulfate on u-PA and P05121 production in synovial cultures at 48 h . CONCLUSIONS : Our data suggest that one of the therapeutic effects of glucosamine sulfate is to down-regulate the expressions of u-PA , P05121 , P08253 and P14780 that underlie the destruction of articular cartilage in the early stage of OA , and therefore to delay the joint failure . Serious obstetric complications interact with hypoxia-regulated/vascular-expression genes to influence schizophrenia risk . The etiology of schizophrenia is thought to include both epistasis and gene-environment interactions . We sought to test whether a set of schizophrenia candidate genes regulated by hypoxia or involved in vascular function in the brain ( P31749 , P23560 , O75052 , P36544 , P21964 , Q96EV8 , Q99259 , Q14832 , Q99466 , Q02297 , O43272 , P49798 , P01375 ) interacted with serious obstetric complications to influence risk for schizophrenia . A family-based study of transmission disequilibrium was conducted in 116 trios . Twenty-nine probands had at least one serious obstetric complication ( OC ) using the McNeil-Sjostrom Scale , and many of the OCs reported were associated with the potential for fetal hypoxia . Analyses were conducted using conditional logistic regression and a likelihood ratio test ( LRT ) between nested models was performed to assess significance . Of the 13 genes examined , four ( P31749 ( three SNPs ) , P23560 ( two SNPs ) , Q96EV8 ( one SNP ) and Q14832 ( one SNP ) ) showed significant evidence for gene-by-environment interaction ( LRT P-values ranged from 0.011 to 0.037 ) . Although our sample size was modest and the power to detect interactions was limited , we report significant evidence for genes involved in neurovascular function or regulated by hypoxia interacting with the presence of serious obstetric complications to increase risk for schizophrenia . Inhibitory effect of selective serotonin reuptake inhibitors on the vesicular monoamine transporter 2 . The neuronal vesicular monoamine transporter ( Q05940 ) is the target molecule of action of some psychostimulants , such as methamphetamine and 3,4-methylenedioxymethamphetamine ( DB01454 ) . The present study examined the effect of antidepressants , such as selective serotonin reuptake inhibitors ( SSRIs ) , on Q05940 activity by measuring adenosine triphosphate-dependent [(3)H]dopamine uptake into synaptic vesicles prepared from rat striatum . SSRIs , fluoxetine , paroxetine , and fluvoxamine , inhibited vesicular [(3)H]dopamine uptake in vitro . The rank order of potency was reserpine >> fluoxetine , paroxetine > fluvoxamine , methamphetamine > DB01454 . Moreover , kinetic analysis revealed that inhibition by reserpine , a typical Q05940 inhibitor , was uncompetitive , decreasing maximum velocity and affinity for dopamine . Inhibition by fluoxetine was noncompetitive , only decreasing maximum velocity for dopamine . These results suggest that fluoxetine inhibited the activity of Q05940 by a mechanism different from that of reserpine and did not directly interact with the active site of Q05940 . 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 . 5-HT₄ receptor stimulation leads to soluble AβPPα production through P14780 upregulation . Serotonin 4 ( Q13639 ) receptor signaling does not only have the physiological function of improving cognition , but might also be helpful in the therapy of Alzheimer 's disease ( AD ) through regulation of the production of soluble amyloid-β protein precursor alpha ( sAβPPα ) . To analyze the relationship between Q13639 receptor signaling and sAβPPα production , we stably transfected H4 cells with AβPP and Q13639 receptor ( H4/AβPP/ Q13639 cells ) . We found that 24-h incubation with the Q13639 receptor agonist RS-67333 upregulates matrix metalloproteinase-9 ( P14780 ) . Furthermore , P14780 overexpression enhanced sAβPPα levels , whereas knockdown with P14780 siRNA decreased sAβPPα levels . When RS-67333 was injected for 10 days in Tg2576 mice , a model of amyloid-β peptide ( Aβ ) deposition , there was an increase in hippocampal levels of sAβPPα , C-terminal fragment α , and P14780 , as well as a decrease in hippocampal senile plaque number and levels of the 40 amino acid peptide , Aβ40 . Taken all together , these experiments demonstrate that Q13639 receptor stimulation induces expression of P14780 which cleaves AβPP through α-secretase-like activity , leading to an increase of sAβPPα levels and a reduction of Aβ load . " DB00945 resistance " in ischemic stroke : insights using short thrombelastography . AIMS : DB00945 achieves its antithrombotic effect through inactivation of cyclo-oxygenase ( P36551 ) -1 , thereby preventing generation of thromboxane (TX)A2 from arachidonic acid ( AA ) . The reported prevalence of aspirin " resistance " varies significantly and is usually based on platelet function tests ( PFTs ) that use AA-induced platelet reactivity as a surrogate measure of the effect of aspirin , rather than specific assessment of its effect on its therapeutic target ( ie , P23219 inhibition ) . The reported rates are not only assay specific but also condition specific , with particularly high rates ( up to 70 % ) previously reported in the stroke population . We investigated whether pharmacological responses to aspirin can be reliably determined from a functional test of AA-induced whole-blood clotting . METHODS AND RESULTS : A prospective study included 35 patients admitted with ischemic stroke and commenced on 300 mg aspirin . AA-induced whole-blood clotting was measured using short thrombelastography , a previously extensively validated near-patient DB00522 . Serum TXB2 and inflammatory biomarkers were also measured . The prevalence of apparent aspirin resistance measured using AA was high ( range from 49 % to 67 % ) . However , serum [ TXB2 ] was consistently low , thereby confirming adequate inhibition of P23219 by aspirin . Mean inflammatory biomarker levels were elevated throughout . CONCLUSION : This study demonstrates that although P23219 activity is adequately and consistently suppressed by aspirin in stroke patients , this effect is not reliably indicated by whole-blood clotting in response to AA . These data help to explain why the reported prevalence of aspirin resistance in stroke from studies employing AA-induced platelet reactivity is high and cast doubt on the veracity of such reports . 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 . 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 . Role of transient receptor potential P01024 in P01375 -enhanced calcium influx in human airway myocytes . Previous studies have suggested that the proinflammatory cytokine , P01375 , contributes to airway hyperresponsivness by altering airway smooth muscle ( P17405 ) Ca(2+) responses to agonist stimulation . The present study examined the effects of P01375 on Ca(2+) influx pathways in cultured human P17405 cells ( HASMCs ) . Proteins encoded by the transient receptor potential ( TRP ) gene family function as channels through which receptor-operated and store-operated Ca(2+) entry ( SOCE ) occur . In the present study , the presence of P48995 , Q13507 , Q9UBN4 , Q9UL62 , and Q9Y210 mRNA and protein expression was confirmed in cultured HASMCs using RT-PCR and Western blot analysis . P01375 treatment significantly increased Q13507 mRNA and protein levels in HASMCs as well as SOCE . P01375 treatment also increased both the peak and plateau intracellular Ca(2+) concentration responses in HASMCs elicited by acetylcholine and bradykinin . The effects of P01375 treatment on SOCE and agonist-induced intracellular Ca(2+) concentration responses were attenuated using small interfering RNA transfection , which knocked down Q13507 expression . Thus , in inflammatory airway diseases , P01375 treatment may result in increased myocyte activation due to altered Ca(2+) influx pathways . These results suggest that Q13507 may be an important therapeutic target in inflammatory airway diseases such as asthma and chronic obstructive pulmonary disease . 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 .

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Tubulin β-III : a novel immunohistochemical marker for intrahepatic peripheral cholangiocarcinoma . AIMS : Our recent proteomic study identified tubulin β-III ( Q13509 ) as a potential tissue marker for intrahepatic cholangiocarcinomas ( CCs ) . This validation study was conducted to see whether or not Q13509 can serve as a novel immunohistochemical marker for peripheral CCs , using a large cohort ( n = 197 ) covering various liver tumours and premalignant conditions . METHODS AND RESULTS : Immunostaining using a monoclonal antibody demonstrated Q13509 expression in 14/28 cases of peripheral CCs ( 50 % ) , while its expression was significantly less common in perihilar CCs ( 6/40 , 15 % ) ( P = 0.002 ) . No significant difference was identified in clinicopathological features between Q13509 -positive and -negative cases . Q13509 expression was entirely negative in hepatocellular carcinomas , biliary premalignant lesions ( i.e. , biliary intraepithelial neoplasias , intraductal papillary neoplasms ) , peribiliary gland hamartomas ( bile duct adenomas ) , and non-neoplastic biliary epithelium . Q13509 expression was only focally noted in 2/12 cases of mixed hepatocellular and cholangiocarcinomas ( < 10 % of cancer cells ) . Compared with other biliary ( CK7 and CK19 ) and malignant markers ( p53 and P15941 ) , Q13509 was less sensitive but more specific for peripheral CCs . Q13509 was also expressed in 40 % of metastatic colorectal or breast cancers . CONCLUSIONS : This study revealed that Q13509 is a moderately sensitive and highly specific tissue marker for discriminating peripheral CCs from other primary liver tumours . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . CpG-DNA-specific activation of antigen-presenting cells requires stress kinase activity and is preceded by non-specific endocytosis and endosomal maturation . Unmethylated CpG motifs in bacterial DNA , plasmid DNA and synthetic oligodeoxynucleotides ( CpG ODN ) activate dendritic cells ( DC ) and macrophages in a P25942 - P29965 -independent fashion . To understand the molecular mechanisms involved we focused on the cellular uptake of CpG ODN , the need for endosomal maturation and the role of the stress kinase pathway . Here we demonstrate that CpG-DNA induces phosphorylation of Jun N-terminal kinase kinase 1 ( P45985 / P54764 / P45985 ) and subsequent activation of the stress kinases P45983 /2 and p38 in murine macrophages and dendritic cells . This leads to activation of the transcription factor activating protein-1 ( AP-1 ) via phosphorylation of its constituents c-Jun and P15336 . Moreover , stress kinase activation is essential for CpG-DNA-induced cytokine release of tumor necrosis factor alpha ( TNFalpha ) and interleukin-12 ( IL-12 ) , as inhibition of p38 results in severe impairment of this biological response . We further demonstrate that cellular uptake via endocytosis and subsequent endosomal maturation is essential for signalling , since competition by non-CpG-DNA or compounds blocking endosomal maturation such as chloroquine or bafilomycin A prevent all aspects of cellular activation . The data suggest that endosomal maturation is required for translation of intraendosomal CpG ODN sequences into signalling via the stress kinase pathway , where p38 kinase activation represents an essential step in CpG-ODN-triggered activation of antigen-presenting cells . Direct oral anticoagulants in acute coronary syndrome . Patients with acute coronary syndromes ( ACS ) require a specific antithrombotic therapy in the immediate and the post ACS phase . The current antithrombotic therapy in the acute phase of an ACS combines antiplatelet and anticoagulant drugs in order to reduce ischemic cardiovascular events . In the post ACS phase , dual antiplatelet therapy ( DAPT ; aspirin and a Q9H244 receptor antagonist ) is the current mainstay of antithrombotic treatment and is recommended in the guidelines of the major North American and European clinical cardiology associations ( DB00551 , ACC , and ESC ) . Recently , the addition of rivaroxaban , a low dose oral direct factor Xa inhibitor ( 2.5 mg twice daily ) , to DAPT ( aspirin plus second-generation Q9H244 inhibitor ) showed a significant reduction of cardiovascular and overall mortality in the major phase III clinical trial ATLAS ACS 2 TIMI 51 . This led to the approval of low-dose rivaroxaban in addition to aspirin and clopidogrel by the European Medicines Agency ( P15941 ) in 2013 . Other direct oral anticoagulants ( apixaban , dabigatran etexilate ) have also been assessed in phase II ( dabigatran etexilate ) and phase III ( apixaban ) post ACS clinical trials . In the studied dosing regimens , these drugs failed to show a net clinical benefit in addition to dual antiplatelet therapy . The major clinical phase II and III post ACS studies of direct oral anticoagulants are summarized and discussed in this article along with the concept of long-term anticoagulation for the secondary prevention of ischemic events after ACS and implications for the future of antithrombotic therapy in the current era of third-generation Q9H244 receptor inhibitors ( Prasugrel and DB08816 ) . Role of Cox-2 in vascular inflammation : an experimental model of metabolic syndrome . The objective of this work was to demonstrate the role of P35354 enzyme at the vascular in experimental model of metabolic syndrome . SHR male WKY rats were employed ; they were distributed in 8 groups ( n = 8 each ) : control ( W ) ; W + L : WKY rats receiving 20 mg/kg of lumiracoxib by intraesophageal administration ; SHR ; SHR + L : SHR + 20 mg/kg of lumiracoxib by intraesophageal administration ; DB04173 -Fed Rats ( Q9UID3 ) : WKY rats receiving 10 % ( w/v ) fructose solution in drinking water during all 12 weeks ; Q9UID3 + L : Q9UID3 + 20 mg/kg of lumiracoxib by intraesophageal administration ; DB04173 -Fed Hypertensive Rats ( FFHR ) : SHR receiving 10 % ( w/v ) fructose solution in drinking water during all 12 weeks ; and FFHR + L : FFHR + 20 mg/kg of lumiracoxib by intraesophageal administration . Metabolic variables , blood pressure , morphometric variables , and oxidative stress variables were evaluated ; also P08253 and P14780 ( collagenases ) , P19320 , and NF- κ B by Westernblot or IFI were evaluated . FFHR presented all variables of metabolic syndrome ; there was also an increase in oxidative stress variables ; vascular remodeling and left ventricular hypertrophy were evidenced along with a significant increase in the expression of the mentioned proinflammatory molecules and increased activity and expression of collagenase . DB01283 was able to reverse vascular remodeling changes and inflammation , demonstrating the involvement of P35354 in the pathophysiology of vascular remodeling in this experimental model . 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 . Thrombin receptors and their antagonists : an update on the patent literature . IMPORTANCE OF THE FIELD : Thrombin plays a central role in cardiovascular inflammation . Most of the cellular responses to thrombin are mediated by cell surface protease-activated receptors ( PARs ) . Several preclinical studies indicate that PARs are potential targets for treating cardiovascular diseases such as thrombosis , atherosclerosis and restenosis . Among PARs , P25116 has emerged as an important therapeutic target . AREAS COVERED IN THIS REVIEW : This review covers recent advances in the development of thrombin receptors antagonists . It is focused on the search for P25116 antagonists as this is at the moment the most promising and attractive target . However , some early promising studies on PAR-3 and -4 antagonists are also reported . WHAT THE READER WILL GAIN : The review has been written in order to give to the reader hints and references that cover , in our opinion , the most interesting and/or promising approaches in this research field . TAKE HOME MESSAGE : Research on P25116 antagonists has finally led to good clinical candidates such as DB05692 ( Schering-Plough ) and E-5555 ( Eisai Co. ) . Clinical trials clearly demonstrate that development of PAR1 antagonists is not only possible but most likely will lead to development of antiplatelet drugs as well as of drugs useful for the treatment of inflammatory , proliferative and neurodegenerative diseases . 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 . DB06594 in the treatment of major depressive disorder : an assessment of benefits and risks . DB06594 ( AGM ) was approved for the treatment of major depressive disorder ( MDD ) in adults by the European Medicines Agency ( P15941 ) in February 2009 . It is an analogue of melatonin and features a unique pharmacodynamic profile with agonism on both types of melatonergic receptors ( MT1/ P02795 ) and antagonism at serotonergic P28335 receptors . There is , however , an ongoing debate regarding the efficacy and safety of this novel antidepressant agent , originally evoked by claims of a significant publication bias underlying the assessment of AGM being an effective antidepressant . Indeed , two recent comprehensive metaanalyses of published and unpublished clinical trials found evidence for a relevant publication bias . However , due to its statistically significant advantage over placebo based on the results of these metaanalyses AGM must be referred to as an effective antidepressant agent in the acute phase of MDD . However , the effect sizes of AGM in the treatment of MDD were evaluated as being small in comparison to other antidepressant agents . In addition , there is insufficient evidence for the efficacy of AGM in relapse prevention of MDD . Apart from efficacy issues , AGM appears to have the potential to exhibit severe hepatotoxicity ( the P15941 has identified AGM-associated " hepatotoxic reactions " as a new safety concern in September 2013 ) that is currently poorly understood . Considering these aspects , it seems inappropriate to evaluate AGM as an antidepressant agent of first choice . Nevertheless , its unique mechanism of action with particular sleep modulating effects may represent a specific treatment strategy for patients with particular characteristics ; further studies with thorough characterization of patients are needed to test this hypothesis . Insights into the roles of cyclophilin A during influenza virus infection . P62937 ( CypA ) is the main member of the immunophilin superfamily that has peptidyl-prolyl cis-trans isomerase activity . CypA participates in protein folding , cell signaling , inflammation and tumorigenesis . Further , CypA plays critical roles in the replication of several viruses . Upon influenza virus infection , CypA inhibits viral replication by interacting with the M1 protein . In addition , CypA is incorporated into the influenza virus virions . Finally , DB00091 ( DB00091 ) , the main inhibitor of CypA , inhibits influenza virus replication through CypA-dependent and -independent pathways . This review briefly summarizes recent advances in understanding the roles of CypA during influenza virus infection . Nutritional correlates of acute respiratory infections . During October 1992 to June 1993 in eight villages covered by the Primary Health Center Machhra in Meerut District , India , interviews with mothers and examinations of 1600 children aged less than 5 years ( under-fives ) were conducted to examine the relationship between acute respiratory infection ( Q9Y4X5 ) and malnutrition . 42.25 % of all children had an Q9Y4X5 within the last 15 days . Most ARIs ( 73.4 % ) were considered mild ( cough and cold with no pneumonia ) . Pneumonia accounted for 19.5 % of all Q9Y4X5 cases , which were considered moderate . The remaining Q9Y4X5 cases were severe ( severe and very severe pneumonia ) . 57.5 % of all children suffered from protein energy malnutrition ( P15941 ) . 78.6 % of children aged 12-14 months had P15941 . Q9Y4X5 was more common among malnourished children than well-nourished children ( 52.2 % vs. 28.8 % ; p 0.001 ) . The incidence of Q9Y4X5 increased as the nutritional status deteriorated ( p 0.05 ) . It also increased as the midarm circumference decreased ( p 0.001 ) . These findings confirm the synergistic action between malnutrition and infection , in this case Q9Y4X5 . Malnourished children suffer considerable impairment in immunity , especially cellular immunity , which makes them more prone to Q9Y4X5 . These findings reinforce the need to strengthen the quality , quantity , and accessibility of nutritional services , particularly promotion of breast feeding and vitamin A supplementation . Enhancement of auranofin-induced apoptosis in MCF-7 human breast cells by selenocystine , a synergistic inhibitor of thioredoxin reductase . P10599 system plays an important role in regulation of intracellular redox balance and various signaling pathways . P30044 ( TrxR ) is overexpressed in many cancer cells and has been identified as a potential target of anticancer drugs . DB00995 ( AF ) is potent TrxR inhibitor with novel in vitro and in vivo anticancer activities . Selenocystine ( SeC ) is a nutritionally available selenoamino acid with selective anticancer effects through induction of apoptosis . In the present study , we demonstrated the synergistic effects and the underlying molecular mechanisms of SeC in combination with AF on MCF-7 human breast cancer cells . The results showed that SeC and AF synergistically inhibited the cancer cell growth through induction of ROS-dependent apoptosis with the involvement of mitochondrial dysfunction . DNA damage-mediated p53 phosphorylation and down-regulation of phosphorylated AKT and P29323 also contributed to cell apoptosis . Moreover , we demonstrated the important role of TrxR activity in the synergistic action of SeC and AF . Taken together , our results suggest the strategy to use SeC and AF in combination could be a highly efficient way to achieve anticancer synergism by targeting TrxR . 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 . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . DB06594 in depression . INTRODUCTION : DB06594 is a relatively new antidepressant with a mechanism of action that is different from other antidepressants : it is a melatonergic agonist and a P28335 antagonist . It is an effective treatment for depression , with relatively mild side effects . It may be a valuable pharmacological alternative in the clinical approach on depression . AREAS COVERED : The literature about agomelatine has been comprehensively reviewed . DB06594 's efficacy , safety and tolerability are reviewed based on the studies undertaken in patients with major depressive disorder ( MDD ) and bipolar disorder ( BPD ) . EXPERT OPINION : DB06594 has shown an antidepressant effect in preclinical models , and the results of a large-scale clinical trial program , conducted in MDD , indicate both an antidepressant activity and a favorable tolerability profile . DB06594 has no discontinuation syndrome , sexual discomfort is rare , and it is generally weigh neutral . The drug appears to be relatively safe in case of overdose . However , some cases of elevated hepatic transaminases are reported during treatment . As agomelatine has a mechanism of action that differs from other agents , it may represent a valuable additional treatment option in those patients who do not respond fully or who do not tolerate the side effects of other antidepressants . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . 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 ) . 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 . P25116 genotype influences platelet aggregation and procoagulant responses in patients with coronary artery disease prior to and during clopidogrel therapy . Genetic variations of the protease-activated receptor-1 ( P25116 ) have been associated with platelet receptor density and linked to thrombin receptor-activating peptide ( TRAP ) -induced phenotypes of platelet aggregation and P16109 expression . We investigated whether the P25116 intervening sequence-14 A > T dimorphism influences platelet procoagulant activity . We also determined whether the Q9H244 antagonist clopidogrel could offset any observed functional polymorphism of the P25116 receptor by inhibiting Q9H244 -mediated amplification of TRAP-induced responses . We studied 54 patients listed for elective percutaneous coronary intervention assessing TRAP-induced platelet aggregation and markers of procoagulant activity . Platelet responses were measured at baseline , 4 h post clopidogrel 300 mg , and 10 and 28 days following clopidogrel 75 mg daily . Each patient was genotyped for the P25116 intervening sequence-14 A/T dimorphism . Increased platelet aggregation and procoagulant responses were observed with P25116 A allele homozygotes . DB00758 significantly inhibited these platelet responses regardless of P25116 genotype , but did not offset the hyper-reactivity associated with the A/A homozygotes . We conclude that a common sequence variation within the P25116 gene influences TRAP-induced platelet procoagulant activity as well as aggregation . Higher platelet reactivity associated with P25116 IVSn-14 A allele homozygotes persists despite clopidogrel therapy . These individuals may be at higher risk of thromboembolic events and may require additional anti-platelet medication . c-Jun activation-dependent tumorigenic transformation induced paradoxically by overexpression or block of S-adenosylmethionine decarboxylase . All mammalian cells absolutely require polyamines ( putrescine , spermidine , and spermine ) for growth . Here we show that the overexpression of cDNA for S-adenosylmethionine decarboxylase ( P17707 ) , the main regulatory enzyme in the biosynthesis of higher polyamines , induces transformation of rodent fibroblasts when expressed in the sense or the antisense orientation . Both transformants were able to induce invasive tumors in nude mice . Neither transformation was associated with activation of the mitogen-activated protein kinases Erk1 and Erk2 . Instead , the DB00118 DC sense , but not antisense , transformants displayed constitutive activation of the c-Jun NH(2)-terminal kinase ( JNK ) pathway . However , both transformations converged on persistent phosphorylation of endogenous c-Jun at Ser73 . The phenotype of the P17707 sense transformants was reversed by expression of dominant-negative mutants of P45985 ( P45985 ) , P45983 , and c-Jun ( TAM-67 ) , which were also found to impair cytokinesis . Similarly , TAM-67 reverted the morphology of the P17707 -antisense expressors . This report is the first demonstration of a protein whose overexpression or block of synthesis can induce cell transformation . In addition , we show that the polyamine biosynthetic enzymes require c-Jun activation for eliciting their biological effects .

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interacts_with DB00783?

multiple_choice

[ Pharmacy-clinic medication of the month. DB00952 ( naramig ) ] . DB00952 , launched by Glaxo Wellcome under the trade name Naramig , is a potent and selective agonist of P28222 and P28221 vascular receptors . Available as tablets of 2.5 mg , it is indicated in the acute treatment of migraine , with or without aura . A single oral dose of 2.5 mg naratriptan is characterized by a satisfactory clinical efficacy ( already significant after one hour , maximum after 4 hours and persisting during 24 hours ) , a reduction by half of the recurrence of the migraine crisis within the 24 hours and an excellent tolerance profile . Inhibitory effects of vinpocetine on the progression of atherosclerosis are mediated by Akt/NF-κB dependent mechanisms in apoE-/- mice . BACKGROUND : Recent studies have found additional roles for vinpocetine , a potent phosphodiesterase type I inhibitor , in anti-proliferation and anti-inflammation of vascular smooth muscle cells and cancer cells via different mechanisms . In this study , we attempted to investigate whether vinpocetine protected against atherosclerotic development in apoE(-/-) mice and explore the underlying anti-atherogenic mechanisms in macrophages . METHODOLOGY/PRINCIPAL FINDINGS : Vinpocetine markedly decreased atherosclerotic lesion size in apoE(-/-) mice measured by oil red O . Masson 's trichrome staining and immunohistochemical analyses revealed that vinpocetine significantly increased the thickness of fibrous cap , reduced the size of lipid-rich necrotic core and attenuated inflammation . In vitro experiments exhibited a significant decrease in monocyte adhesion treated with vinpocetine . Further , active P01375 -α , P05231 , monocyte chemoattractant protein-1 and matrix metalloproteinase-9 expression induced by ox-LDL were attenuated by vinpocetine in a dose-dependent manner . Similarly , ox-LDL-induced reactive oxygen species were significantly repressed by vinpocetine . Both western blot and luciferase activity assay showed that vinpocetine inhibited the enhanced Akt , IKKα/β , IκBα phosphorylation and NF-κB activity induced by ox-LDL , and the inhibition of NF-κB activity was partly caused by Akt dephosphorylation . However , knockdown of Q01064 did not affect Akt , IKKα/β and IκBα phosphorylation . CONCLUSIONS : These results suggest that vinpocetine exerts anti-atherogenic effects through inhibition of monocyte adhesion , oxidative stress and inflammatory response , which are mediated by Akt/NF-κB dependent pathway but independent of PDE1 blockade in macrophages . Targeting c-erbB2 and other receptors of the c-erbB family : rationale and clinical applications . The c-erbB family of receptors includes four distinct receptors , namely c-erb B1 , 2 , 3 and 4 ( P00533 , 2 , 3 and 4 , respectively ) . DB00072 ( T ) is a recombinant humanized anti- P04626 monoclonal antibody that binds the extracellular domain of the receptor and blocks intracellular signalling . In clinical studies of T , either alone or in combination with chemotherapy , in P04626 overexpressing metastatic breast cancer patients , a significant benefit was obtained -- improved response rates and survival , when T was combined with chemotherapy . Several trials of adjuvant T , either singly or in combination with chemotherapy , are in progress in early breast cancer patients . DB06366 defines a new class of P04626 inhibitors , " dimerization inhibitors " that block both homo- and hetero-dimerization of P04626 . In preclinical studies pertuzumab is inhibitory to breast , prostate and non small cell lung cancer cell lines , both over and non overexpressing P04626 . In phase I clinical trials pertuzumab has shown activity in a number of human cancers . A phase II program is in progress . Primary brain lymphoma cell turnover differs in patients with and without AIDS : relationships to bcl-2 expression and host cell reaction . Primary central nervous system lymphomas ( PCNSLs ) are more resistant to radiotherapy and chemotherapy in AIDS ( A-PCNSLs ) than in non-AIDS patients ( NA-PCNSLs ) . We investigated 23 A-PCNSLs and 24 NA-PCNSLs . Lymphoma cell kinetics ( i.e. proliferation [ mitotic index , MIB-1 and P12004 labeling indices ] , apoptosis and turnover ) were determined and compared with bcl-2 and Q9NR12 -1 expression , and to the percentage of tumor-infiltrating T-lymphocytes ( T-TILs ) and macrophages . A-PCNSLs showed lower proliferation ( p < 0.005 ) , less apoptosis ( p < 0.0001 ) and slower cell-turnover ( p < 0.0001 ) than NA-PCNSLs . Q9NR12 -1 was detected in 90 % of A-PCNSLs and 5 % of NA-PCNSLs , a finding correlating positively with bcl-2 expression ( p < 0.0007 ) . In contrast , T- Q15399 counts and P01730 /CD8 T- Q15399 ratios were similar in A-PCNSLs and NA-PCNSLs . T- Q15399 counts correlated negatively with proliferation indices ( from p < 0.05 to p < 0.0005 ) in NA-PCNSLs , but not in A-PCNSLs . Macrophage counts correlated positively with apoptosis in both groups . We concluded the following : ( i ) A-PCNSLs are characterized by accumulation of slow-cycling , long-lived cells that might be protected from apoptosis by Q9NR12 -1 induced bcl-2 expression , and independently from the host response ; ( ii ) NA-PCNSLs are characterized by a faster cell turnover associated with an insufficient antiproliferative host response ; and ( iii ) A-PCNSLs and NA-PCNSLs constitute 2 entities with distinctive morphology and different kinetic profiles that could account for different responses to therapy . Cdk5 phosphorylates dopamine D2 receptor and attenuates downstream signaling . The dopamine D2 receptor ( P14416 ) is a key receptor that mediates dopamine-associated brain functions such as mood , reward , and emotion . P12004 -dependent kinase 5 ( Cdk5 ) is a proline-directed serine/threonine kinase whose function has been implicated in the brain reward circuit . In this study , we revealed that the serine 321 residue ( S321 ) in the third intracellular loop of P14416 ( D2i3 ) is a novel regulatory site of Cdk5 . Cdk5-dependent phosphorylation of S321 in the D2i3 was observed in in vitro and cell culture systems . We further observed that the phosphorylation of S321 impaired the agonist-stimulated surface expression of P14416 and decreased G protein coupling to P14416 . Moreover , the downstream DB02527 pathway was affected in the heterologous system and in primary neuronal cultures from p35 knockout embryos likely due to the reduced inhibitory activity of P14416 . These results indicate that Cdk5-mediated phosphorylation of S321 inhibits P14416 function , providing a novel regulatory mechanism for dopamine signaling . Systems biology evaluation of immune responses induced by human host defence peptide LL-37 in mononuclear cells . The immune system is very complex , it involves the integrated regulation and expression of hundreds of proteins . To understand in greater detail how the human host defence immunomodulatory peptide LL-37 interacts with innate immunity , a systems approach was pursued . Polychromatic flow cytometry was employed to demonstrate that within human peripheral blood mononuclear cells , P08571 + monocytes , myeloid and plasmocytoid dendritic cells and T- and B-lymphocytes , all responded to LL-37 , with the differential production of intracellular cytokines . Microarray analyses with P08571 + monocytes indicated the differential expression of 475 genes in response to stimulation with LL-37 . To understand this complex response , bioinformatic interrogation , using InnateDB , of the gene ontology , signalling pathways and transcription factor binding sites was undertaken . Activation of the P25963 /NFkappaB , mitogen-activated protein kinases p38 , P27361 /2 and JNK , and PI3K signalling pathways in response to LL-37 was demonstrated by pathway and ontology over-representation analyses , and confirmed experimentally by inhibitor studies . Computational analysis of the predicted transcription factor binding sites upstream of the genes that were regulated by LL-37 predicted the involvement of several transcription factors including NFkappaB and five novel factors , AP-1 , P05549 , SP-1 , Q01094 , and EGR , which were experimentally confirmed to respond to LL-37 by performing transcription factor array studies on nuclear extracts from LL-37 treated mononuclear cells . These data are discussed as reflecting the integration of several responsive signalling pathways through the involvement of transcription factor complexes in gene expression activated by LL-37 in human mononuclear cells . 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 . A multi-layer inference approach to reconstruct condition-specific genes and their regulation . An important topic in systems biology is the reverse engineering of regulatory mechanisms through reconstruction of context-dependent gene networks . A major challenge is to identify the genes and the regulations specific to a condition or phenotype , given that regulatory processes are highly connected such that a specific response is typically accompanied by numerous collateral effects . In this study , we design a multi-layer approach that is able to reconstruct condition-specific genes and their regulation through an integrative analysis of large-scale information of gene expression , protein interaction and transcriptional regulation ( transcription factor-target gene relationships ) . We establish the accuracy of our methodology against synthetic datasets , as well as a yeast dataset . We then extend the framework to the application of higher eukaryotic systems , including human breast cancer and Arabidopsis thaliana cold acclimation . Our study identified P09758 ( P09758 ) as a target gene for human breast cancer and discovered its regulation by transcription factors CREB , as well as NFkB . We also predict Q99661 is a target gene for ER-/ P04626 - breast cancer and is positively regulated by Q01094 . The predictions were further confirmed through experimental studies . AVAILABILITY : The implementation and detailed protocol of the layer approach is available at http://www.egr.msu.edu/changroup/Protocols/Three-layer % 20approach % 20 to % 20reconstruct % 20condition.html . Does a nonclassical signaling mechanism underlie an increase of estradiol-mediated gonadotropin-releasing hormone receptor binding in ovine pituitary cells ? DB00783 ( E2 ) is the major regulator of P30968 ( GnRHR ) gene expression and number during the periovulatory period ; however , the mechanisms underlying E2 regulation of the P30968 gene remain undefined . Herein , we find that E2 conjugated to BSA ( E2-BSA ) mimics the stimulatory effect of E2 on DB00644 binding in primary cultures of ovine pituitary cells . The time course for maximal DB00644 analog binding was similar for both E2 and E2-BSA . The ability of E2 and E2-BSA to increase DB00644 analog binding was blocked by the estrogen receptor ( ER ) antagonist DB00947 . Also , increased DB00644 analog binding in response to E2 and the selective P03372 agonist propylpyrazole triol was blocked by expression of a dominant-negative form of P03372 ( L540Q ) . Thus , membrane-associated P03372 is the likely candidate for mediating E2 activation of the P30968 gene . As DB02527 response element binding protein ( CREB ) is an established target for E2 activation in gonadotrophs , we next explored a potential role for this protein as an intracellular mediator of the E2 signal . Consistent with this possibility , adenoviral-mediated expression of a dominant-negative form of CREB ( A-CREB ) completely abolished the ability of E2 to increase DB00644 analog binding in primary cultures of ovine pituitary cells . Finally , the presence of membrane-associated E2 binding sites on ovine pituitary cells was demonstrated using a fluorescein isothiocyanate conjugate of E2-BSA . We suggest that E2 regulation of GnRHR number during the preovulatory period reflects a membrane site of action and may proceed through a nonclassical signaling mechanism , specifically a CREB-dependent pathway . Internalization of OspA in rsCD14 complex and aggregated forms . Although the spirochetal protein OspA is capable of stimulating immune cells in a P08571 - and O60603 -dependent manner , little is known about how O60603 receptor complex ligands , such as OspA , are handled by the cell once delivered . We examine here the internalization of the fluorescently derivatized forms of both the full length DB00045 delivered as a recombinant soluble P08571 ( rsCD14 ) complex and the corresponding lipohexapeptide given to the cells as an aggregate . Both forms of OspA are internalized in a similar manner to acetylated low density lipoprotein ( AcLDL ) , a scavenger receptor ligand . Acetylated low density lipoprotein is capable of competing for internalization with OspA even when OspA is delivered as a rsCD14 complex . We observe co-localization of OspA with lysosomes but not with the Golgi complex . These phenomena are similar between RAW264.7 macrophages and endothelial cells but change drastically when the cells are deprived of serum . Upon serum starvation , OspA shows some localization to the Golgi apparatus whereas the lipohexapeptide remains on the cell surface . Inhibition of internalization of OspA via treatment with cytochalasin D or of the lipohexapeptide via serum starvation does not interfere with P01375 induction activity , consistent with signalling from the cell surface . Proliferative response of mantle cell lymphoma cells stimulated by P25942 ligation and P05112 . Mantle cell lymphoma ( Q8WXI8 ) is a tumor of intermediate-size , IgM+ , IgD+ B cells derived from the mantle zone of the germinal center . Little is known about its specific immunologic features or responsiveness to T cell-derived signals . In this work , we evaluated the proliferation and cell cycle properties of freshly isolated Q8WXI8 cells after P25942 ligation , in the absence and presence of interleukin 4 ( P05112 ) . In each Q8WXI8 case examined , there was a marked growth-enhancing effect of these two stimuli characterized by improved viability , augmented expression of Ki-67 , and induction of the proliferating cell nuclear antigen ( P12004 ) . P12004 D1 was expressed throughout the cell cycle in Q8WXI8 cells induced to enter S phase . From these investigations , we conclude that the biology of Q8WXI8 B lymphocytes is affected by CD154 ( P29965 ) and P05112 , two signals usually provided by P01730 + T cells . The capacity to manipulate the activation and cell cycle state of Q8WXI8 cells by these specific immunological stimuli may be exploited to confer susceptibility to chemotherapy agents and develop novel therapies in this disease . DB06366 plus trastuzumab in combination with standard neoadjuvant anthracycline-containing and anthracycline-free chemotherapy regimens in patients with P04626 -positive early breast cancer : a randomized phase II cardiac safety study ( TRYPHAENA ) . BACKGROUND : DB06366 ( P ) combined with trastuzumab ( H ) -based chemotherapy improves efficacy in early and advanced P04626 -positive breast cancer . We assessed the tolerability , with particular focus on cardiac safety , of H and P with chemotherapy in the neoadjuvant treatment of P04626 -positive early breast cancer . PATIENTS AND METHODS : In this multicenter , open-label phase II study , patients with operable , locally advanced , or inflammatory breast cancer were randomized 1 : 1 : 1 to receive six neoadjuvant cycles q3w ( Arm A : 5-fluorouracil , epirubicin , cyclophosphamide [ FEC ] + H + P ×3 → docetaxel [ T ] + H + P ×3 ; Arm B : FEC ×3 → T + H + P ×3 ; Arm C : T + carboplatin + H [TCH]+P ×6 ) . pCR was assessed at surgery and adjuvant therapy given to complete 1 year of H . RESULTS : Two hundred twenty-five patients were randomized . During neoadjuvant treatment , two patients ( 2.7 % ; Arm B ) experienced symptomatic left ventricular systolic dysfunction ( LVSD ) and 11 patients ( Arm A : 4 [ 5.6 % ] ; Arm B : 4 [ 5.3 % ] ; Arm C : 3 [ 3.9 % ] ) had declines in left ventricular ejection fraction of ≥10 % points from baseline to < 50 % . Diarrhea was the most common adverse event . pCR ( ypT0/is ) was reported for 61.6 % ( Arm A ) , 57.3 % ( Arm B ) , and 66.2 % ( Arm C ) of patients . CONCLUSION : The combination of P with H and standard chemotherapy resulted in low rates of symptomatic LVSD . Q14201 tumor suppressor gene promoter demethylation , histone modification and cell cycle arrest by genistein in renal cancer . Q14201 / Q14201 /APRO4 has been reported to be a tumor suppressor gene in some malignancies . It constitutes important negative regulatory mechanism for Src-mediated signaling , a negative regulator of the cell cycle and inhibits transcription factor Q01094 . We report that Q14201 is downregulated in renal cancer and that the mechanism of inactivation is through promoter hypermethylation . Quantitative real-time polymerase chain reaction ( PCR ) showed that Q14201 was downregulated in cancer tissues and cells . DB01645 and DB01262 ( 5Aza-C ) induced Q14201 messenger RNA ( mRNA ) expression in A498 , ACHN and P29320 -293 renal cell carcinoma ( RCC ) cell lines . Bisulfite-modified PCR and DNA sequencing results showed complete methylation of Q14201 promoter in tumor samples and cancer cell lines . DB01645 and 5Aza-C treatment significantly decreased promoter methylation , reactivating Q14201 expression . Chromatin immunoprecipitation assay revealed that genistein and 5Aza-C increased levels of acetylated histones 3 , 4 , 2H3K4 , 3H3K4 and RNA polymerase II at the Q14201 promoter indicative of active histone modifications . Enzymatic assays showed genistein and 5Aza-C decreased DNA Methyltransferase , methyl-CpG-binding domain 2 activity and increased O60235 activity . Cell cycle and 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide cell proliferation assays showed that genistein has antiproliferative effect on cancer cell growth through induction of cell cycle arrest . This is the first report to show that Q14201 is epigenetically silenced in RCC and can be reactivated by genistein-induced promoter demethylation and active histone modification . DB01645 had similar effects to that of 5Aza-C , which is a potent demethylating agent with high toxicity and instability . DB01645 being a natural , non-toxic , dietary isoflavone is effective in retarding the growth of RCC cells , making it a promising candidate for epigenetic therapy in renal carcinoma . Dexamethasone reverses adrenalectomy-induced neuronal de-differentiation in midbrain raphe-hippocampus axis . Differentiation leads to specific morphological and biochemical characteristics . We examined whether epigenetic factors ( e.g. , glucocorticoids ) are required to maintain neuronal differentiation in the adult brain . In the midbrain , adrenalectomy ( P10109 ) ( 1-2 wk ) reduced the size of tryptophan hydroxylase ( WH ) -immunoreactive ( IR ) neurons . P10109 rats exposed to short-term ( 24-72-h ) dexamethasone ( ST-DEX ) in the drinking saline ( 10 mg/l ) showed an increase in WH protein , somal area and dendritic size of WH-IR neurons . In the hippocampus , P10109 for 2-3 mo ( long-term ; LT ) reduced Nissl staining , calbindin ( DB09061 ) -IR and P08908 receptor mRNA in the granular cell layer , and the size of the molecular layer and its DB09061 -IR dendrites . Small vimentin ( Vim ) -IR glial cells emerged in the granular layer . ST-DEX after LT- P10109 rapidly induced a recovery of P08908 mRNA , Nissl labeling and DB09061 -IR in the granule cell layer . In the molecular layer , there was an increase in the area and in the number of DB09061 -IR dendrites . Furthermore , the Vim-IR glial cells were enlarged in size and branching . The rate of cell proliferation was studied in these animals . Immunostaining with antibodies against proliferating cell nuclear antigen ( P12004 ) and use of bromouridine argue against enhanced neurogenesis after ST-DEX in LT- P10109 . We propose that glucocorticoids induce and maintain differentiation of serotonergic and DB09061 -IR neurons in the midbrain-hippocampal axis . A neuronotrophic role for the glial P08908 receptor is suggested . [ 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 . 5-hydroxytryptamine stimulates phosphorylation of Q8TCB0 / Q8NFH3 mitogen-activated protein kinase activation in bovine aortic endothelial cell cultures . 5-Hydroxytryptamine ( 5-HT ) is sequestered and released by endothelial cells , acts as an endothelial cell mitogen , promotes the release of nitric oxide ( NO ) , and has been associated with the Q8TCB0 / Q8NFH3 mitogen-activated protein kinase ( MAPK ) cascade . NO also acts as a cell mitogen and promotes signals that culminate in the phosphorylation of MAPK . The aim of this study was to test whether endothelial 5-HT receptors stimulate dual ( tyrosyl- and threonyl- ) phosphorylation of MAPK through a mitogen-activated protein kinase kinase-1 ( MEK-1 ) and P29474 -dependent pathway in bovine aortic endothelial cells ( BAECs ) . As shown by Western blot analysis , 5-HT and the P28222 -selective agonist 5-nonyloxytryptamine ( 5-NOT ) stimulate time- and concentration-dependent ( 0.001-10 microM ) phosphorylation of MAPK in these cells . The agonist-stimulated phosphorylation of MAPK was blocked by the 5-HT1b-receptor antagonist isamoltane ( 0.01-10 p3M ) and the MEK-1 inhibitor PD 098059 ( [ 2-(2'-amino-3'-methoxy-phenyl)-oxanaphthalen-4-one ] ; 0.01-10 microM¿ . The P29474 inhibitor L-N(omega)-iminoethyl-L-ornithine ( L-NIO ; 0.01-10 microM ) failed to block the 1 microM 5-NOT-stimulated responses . Our findings suggest that the 5-HT receptors ( specifically P28222 ) mediate signals to MEK-1 and subsequently to MAPK through an P29474 -independent pathway in BAECs . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . DB04216 arrests G2/M phase and induces caspase-dependent cell death in U937 cells . DB04216 , a natural product derived from grapes , has been shown to prevent carcinogenesis in murine models . We report here that quercetin induces anti-proliferation and arrests G2/M phase in U937 cells . The G2/M phase accumulation was accompanied by an increase in the level of the cyclin B . In contrast , the level of the cyclin D , cyclin E , Q01094 , and Q14209 was marked decreased in quercetin-treated U937 cells . Removal of quercetin from the culture medium stimulates U937 cells to synchronously re-enter the cell cycle , decrease expression level of cyclin B , and increased the expression level of cyclin D and cyclin E . These data demonstrate that quercetin causes reversible G2/M phase arrest , which was related with dramatic changes in the level of cyclin B , cyclin D , and cyclin E. DB04216 -induced down-regulation of cyclin D and cyclin E was associated with suppression of transcriptional levels but not protein stability . In addition , quercetin-treated U937 cells showed DNA fragmentation , increased sub- P55008 population , and generated a 60kDa cleavage product of P98160 -gamma1 in a dose-dependent manner , which were significantly inhibited by z-VAD-fmk . These data clearly indicate that quercetin-induced apoptosis is associated with caspase activation . In summary , the growth inhibition of the quercetin is highly related to cell cycle arrest at the G2/M phase and induction of caspase-dependent apoptosis in human promonocytic U937 cells . 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 . Engineering the response to vascular injury : divergent effects of deregulated Q01094 expression on vascular smooth muscle cells and endothelial cells result in endothelial recovery and inhibition of neointimal growth . P01375 -alpha ( P01375 ) is expressed locally in the vessel wall after angioplasty and induces growth arrest and apoptosis in endothelial cells ( ECs ) , thereby delaying reendothelialization . Prior studies have shown that direct antagonism of P01375 , using a systemically administered soluble receptor , can enhance endothelial recovery and reduce neointimal thickening . These studies have also shown that downregulation of the transcription factor Q01094 was a key mechanism of P01375 's effect on ECs . We now show that Ad- Q01094 overexpression at sites of balloon injury accelerates functional endothelial recovery , consistent with the prior in vitro findings . Moreover these studies also reveal divergent effects of P01375 and overexpression of Q01094 on ECs versus VSMCs . P01375 exposure of VSMCs had no affect on proliferation or apoptosis , in contrast to the effect seen in ECs . In Ad- Q01094 -transduced VSMCs , however , P01375 -induced marked apoptosis in contrast to the survival effect seen in ECs . Finally , these studies suggest that differential activation of NF-kappaB may play a key role in mediating these opposing effects . Nuclear translocation and transcriptional activity of NF-kappaB was markedly attenuated in Ad- Q01094 -transduced VSMCs , whereas it remained active in similarly treated ECs when the cells were exposed to P01375 . These studies reveal that overexpression of Ad- Q01094 primes VSMCs to P01375 -induced apoptosis . Furthermore , Q01094 potentiates VSMC death by blocking antiapoptotic signaling pathway through inhibition of NF-kappaB activation . The divergent responses of VSMCs and ECs to Q01094 overexpression provide unique therapeutic possibilities : simultaneously targeting the cell cycle of two different cell types , within same tissue microenvironment resulting in opposite and biologically complimentary effects . 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 . Differential regulation of human antigen-specific Th1 and Th2 lymphocyte responses by isozyme selective cyclic nucleotide phosphodiesterase inhibitors . Our study explores the relative efficacy of phosphodiesterase ( PDE ) inhibitors on antigen-specific Th1 and Th2 clonal responses . Proliferative responses for both phenotypes were down-regulated by the DB05876 inhibitor , rolipram , but not the PDE3 inhibitor , siguazodan . The Th2 clones were more sensitive than the Th1 clones to DB05876 inhibition ( P < .05 at 10 and 100 microM rolipram ) . The addition of 1 microM of the adenylyl cyclase activator , isoproterenol , significantly decreased both the EC50 and IC50 of rolipram in both phenotypes ( P < .05 ) . Gene expression for interleukin-4 , interleukin-5 , or interferon-gamma , assessed by reverse transcription-polymerase chain reaction , was down-regulated by the DB05876 inhibitor , but not the PDE3 inhibitor , in each respective clone . Cytokine protein secretion paralleled the results of reverse transcription-polymerase chain reaction for P05112 and interferon-gamma ( P < .01 for each ) . No differential efficacy on cytokine generation parameters between T helper phenotypes was apparent . DB01954 treatment significantly elevated intracellular cyclic AMP ( adenosine 3',5'-cyclic monophosphate ) in clonal T cells ( P < .01 for Th1 or Th2 clones ) ; these elevations were consistently greater in the Th2 clones ( P < .05 ) . Finally , Th1 cells showed reduced gene expression for the Q08493 isoform and a lack of gene expression for the Q08499 isoform by reverse transcription-polymerase chain reaction , compared to the Th2 cells . These data demonstrate the potent immunomodulatory efficacy of DB05876 inhibition on antigen-specific T cell clones . The enhanced sensitivity of Th2 cells to DB05876 inhibition may be due , in part , to the differential expression of DB05876 isoforms between Th1 and Th2 cells . Regulation of dihydrofolate reductase gene expression and E2F components in human diploid fibroblasts during growth and senescence . The induction of dihydrofolate reductase ( P00374 ) , a key enzyme in DNA biosynthesis that is induced just before the onset of S phase , is markedly attenuated in senescent human fibroblasts ( Pang and Chen , 1994 , J. Cell. Physiol. , 160:531-538 ) . Footprinting analysis of the 365 bp promoter region of the human P00374 gene ( -381 to -17 ) indicated that nuclear proteins bind to a cluster of cis-elements , including two overlapping E2F binding sequences , two Sp1 sites , and one Yi sequence . Gel mobility shift assays were performed to assess the role of each cis-element in the regulation of P00374 gene expression . We found that 1 ) Sp1 binding activity was constitutively expressed throughout the cell cycle in early passage and senescent cells ; 2 ) Yi binding activity was undetectable in both early passage and senescent cells ; and 3 ) E2F binding activity was serum-inducible , senescence-dependent , and prominent in presenescent cells but strikingly diminished in senescent cells . Northern blot analysis of the expression of E2F and DP family members showed that the Q01094 , Q16254 , and Q15329 mRNA was growth- and senescence-dependent , whereas O00716 , DP-1 , and DP-2 expression was constitutive and senescence-independent . In contrast , Q14209 mRNA was not detectable in IMR-90 or WI-38 human fibroblasts . Western blot analysis showed that among the E2F-associated proteins , the expression of Q01094 , cyclin A , and cyclin B but not P28749 was cell cycle- and senescence-dependent . A nuclear extract mixing experiment suggested that an inhibitory factor may further reduce E2F binding activity in senescent cells . Blockade of interleukin-6 signalling with siltuximab enhances melphalan cytotoxicity in preclinical models of multiple myeloma . Signalling through the interleukin ( IL ) -6 pathway induces proliferation and drug resistance of multiple myeloma cells . We therefore sought to determine whether the P05231 -neutralizing monoclonal antibody siltuximab , formerly CNTO 328 , could enhance the activity of melphalan , and to examine some of the mechanisms underlying this interaction . DB09036 increased the cytotoxicity of melphalan in KAS-6/1 , Q16352 -6 , ANBL-6 , and RPMI 8226 human myeloma cell lines ( HMCLs ) in an additive-to-synergistic manner , and sensitized resistant RPMI 8226.LR5 cells to melphalan . These anti-proliferative effects were accompanied by enhanced activation of drug-specific apoptosis in HMCLs grown in suspension , and in HMCLs co-cultured with a human-derived stromal cell line . DB09036 with melphalan enhanced activation of caspase-8 , caspase-9 , and the downstream effector caspase-3 compared with either of the single agents . This increased induction of cell death occurred in association with enhanced Bak activation . Neutralization of P05231 also suppressed signalling through the phosphoinositide 3-kinase/Akt pathway , as evidenced by decreased phosphorylation of Akt , P08133 S6 kinase and Q13541 . Importantly , the siltuximab/melphalan regimen demonstrated enhanced anti-proliferative effects against primary plasma cells derived from patients with myeloma , monoclonal gammopathy of undetermined significance , and amyloidosis . These studies provide a rationale for translation of siltuximab into the clinic in combination with melphalan-based therapies . Hinokitiol Negatively Regulates Immune Responses through Cell Cycle Arrest in Concanavalin A-Activated Lymphocytes . Autoimmune diseases are a group of chronic inflammatory diseases that arise from inappropriate inflammatory responses . Hinokitiol , isolated from the wood of Chamaecyparis taiwanensis , engages in multiple biological activities . Although hinokitiol has been reported to inhibit inflammation , its immunological regulation in lymphocytes remains incomplete . Thus , we determined the effects of hinokitiol on concanavalin A- ( ConA- ) stimulated T lymphocytes from the spleens of mice . In the present study , the MTT assay revealed that hinokitiol ( 1-5 μM ) alone did not affect cell viability of lymphocytes , but at the concentration of 5 μM it could reduce ConA-stimulated T lymphocyte proliferation . Moreover , propidium iodide ( PI ) staining revealed that hinokitiol arrested cell cycle of T lymphocytes at the G0/ P55008 phase . Hinokitiol also reduced interferon gamma ( IFN-γ ) secretion from ConA-activated T lymphocytes , as detected by an ELISA assay . In addition , hinokitiol also downregulated cyclin D3 , Q01094 , and Cdk4 expression and upregulated P38936 expression . These results revealed that hinokitiol may regulate immune responses . In conclusion , we for the first time demonstrated that hinokitiol upregulates P38936 expression and attenuates IFN-γ secretion in ConA-stimulated T lymphocytes , thereby arresting cell cycle at the G0/ P55008 phase . In addition , our findings also indicated that hinokitiol may provide benefits to treating patients with autoimmune diseases . Human P30939 receptor-stimulated [35S]GTPgammaS binding : correlation with inhibition of guinea pig dural plasma protein extravasation . To determine the potency and efficacy of P30939 receptor ligands , a [35S]GTPgammaS binding assay was developed and optimized for the human P30939 receptor . Compounds which are known to be effective in the abortive treatment of migraine were tested for efficacy and potency in this assay . DB00952 , sumatriptan , zolmitriptan , and rizatriptan all had agonist activity . The P30939 receptor ligand LY334370 ( 4-fluoro-N-[3-(1-methyl-4-piperidinyl)-1H-indol-5-yl]-benzamide ) was the most potent compound tested with an EC50 of 2.13 +/- 0.15 nM . LY302148 ( 5-fluoro-3-[1-[2-(1-methyl-1H-pyrazol-4-yl)ethyl]-4-piperidinyl]-1H-ind ole ) , methysergide , LY306258 ( 3-dimethylamino-2,3,4,9-tetrahydro-1H-carbazol-6-ol ) , dihydroergotamine ( DHE ) , L-694,247 and CP-122,288 were also investigated for potency and efficacy . There was a statistically significant correlation between the pEC50 for the stimulation of [35S]GTPgammaS binding and the pID50 for the inhibition of trigeminal nerve-stimulated dural plasma protein extravasation in the guinea pig . In the course of these studies , it was found that the purportedly selective P28221 receptor antagonist GR127935 inhibited P30939 receptor-stimulated [35S]GTPgammaS binding with a Ki of 39.6 +/- 9.5 nM . These studies demonstrate that P30939 receptor-mediated stimulation of [35S]GTPgammaS binding in a clonal cell system is a reproducible , high throughput assay that is predictive of an in vivo model of P30939 receptor activation . 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 . Chimaeric gonadotropin-releasing hormone ( DB00644 ) peptides with improved affinity for the catfish ( Clarias gariepinus ) P30968 . The gonadotropin-releasing hormone ( DB00644 ) receptor in catfish differs from its mammalian counterparts in showing a very low affinity for the hypothalamic DB00644 form [ i.e. catfish DB00644 ( cfGnRH ) ] and a very high affinity for the highly conserved mesencephalic DB00644 , chicken DB00644 -II ( cGnRH-II ) . In the present study we investigated the molecular interactions between ligand and receptor involved in determining the ligand selectivity of the catfish P30968 . Studies on the binding characteristics of the catfish P30968 for cfGnRH and cGnRH-II as well as for mammalian DB00644 ( mGnRH ) and synthetic chimaeric GnRHs , differing at positions 5 , 7 and 8 , revealed that the low affinity of the catfish receptor for cfGnRH can be improved by replacing DB00149 (7) by a tryptophan residue and/or DB00174 (8) by either a tyrosine or an arginine residue . Testing cfGnRH and cGnRH-II as well as mGnRH and the chimaeric GnRHs on DB00128 (304) --> Ala , DB00128 (304) --> DB00142 and DB00128 (304) --> DB00174 mutant catfish DB00644 receptors revealed that DB00128 (304) of the catfish receptor mediates the recognition of DB00125 (8) in mGnRH , as well as in the chimaeric peptides [ DB00125 (8)]cfGnRH and [ DB00125 (8)]cGnRH-II , but seems to be less important for the recognition of DB00135 (8) in cGnRH-II . On the basis of these results , a three-dimensional model for the binding of [ DB00125 (8)]cGnRH-II to the catfish P30968 is proposed . Characterization of IRA/IRB hybrid insulin receptors using bioluminescence resonance energy transfer . The insulin receptor ( IR ) is composed of two alpha-chains that bind ligands and two beta-chains that possess an intracellular tyrosine kinase activity . The IR is expressed in cells as two isoforms containing or not exon 11 ( IRB and IRA , respectively ) . Several mRNA studies have demonstrated that the two isoforms are co-expressed in different tissues and in several cancer cells . IRA/IRB hybrid receptors , constituting of an alphabeta-chain from IRA and an alphabeta-chain from IRB , are likely to occur in cells co-expressing both isoforms , but their study has been hampered by the lack of specific tools . In previous work , we used BRET to study IR and P08069 homodimers and heterodimers . Here , we have used BRET to characterize IRA/IRB hybrids . BRET saturation experiments showed that IRA/IRB hybrids are randomly formed in cells . Moreover , by co-transfecting P29320 -293 cells with a luciferase-tagged kinase-dead version of one isoform and a wild-type untagged version of the other isoform , we showed that IRA/IRB hybrids can recruit , upon ligand stimulation , a YFP-tagged intracellular partner . Finally , using BRET , we have studied ligand-induced conformational changes within IRA/IRB hybrids . Dose-response experiments showed that hybrid receptors bind IGF-2 with the same affinity than IRA homodimers , whereas they bind DB01277 with a lower affinity . Altogether , our data indicate that IRA/IRB hybrid receptors can form in cells co-expressing both IR isoforms , that they are capable of recruiting intracellular partners upon ligand stimulation , and that they have pharmacological properties more similar to those of IRA than those of IRB homodimers with regards to IGF-2 . P23458 activates P40763 activity in non-small-cell lung cancer cells and P05231 neutralizing antibodies can suppress P23458 - P40763 signaling . Members of the signal transducer and activator of transcription ( P35610 ) family of transcription factors are potential targets for the treatment and prevention of cancers including non-small-cell lung cancer . P35610 proteins can be phosphorylated and activated by diverse upstream kinases including cytokine receptors and tyrosine kinases . We examined P35610 protein activation in lung cancer cell lines including those with activating mutations in the P00533 and examined upstream kinases responsible for P40763 phosphorylation and activation using small molecules , antibodies , and RNA interference . We found more pronounced P40763 activation in cells with activating P00533 mutations , yet inhibition of P00533 activity had no effect on P40763 activation . Inhibition of P23458 with small molecules or RNA interference resulted in loss of P40763 tyrosine phosphorylation and inhibition of cell growth . An interleukin-6 neutralizing antibody , siltuximab ( CNTO 328 ) could inhibit P40763 tyrosine phosphorylation in a cell-dependent manner . DB09036 could completely inhibit P40763 tyrosine phosphorylation in H1650 cells , and this resulted in inhibition of lung cancer cell growth in vivo . Combined P00533 inhibition with erlotinib and siltuximab resulted in dual inhibition of both tyrosine and serine P40763 phosphorylation , more pronounced inhibition of P40763 transcriptional activity , and translated into combined effects on lung cancer growth in a mouse model . Our results suggest that P23458 is responsible for P40763 activation in lung cancer cells and that indirect attacks on P23458 - P40763 using an P05231 neutralizing antibody with or without P00533 inhibition can inhibit lung cancer growth in lung cancer subsets . 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 . Growth arrest and forced differentiation of human primary glioblastoma multiforme by a novel small molecule . Glioblastoma multiforme is the most common malignant brain tumor in adults , with an average survival of less than one year due to its resistance to therapy . Recent studies reported that GBM initiates from CD133-expressing cancer stem cells ( CSC ) . However , the efficacy of CSC targeting is limited . A newly developed approach in cancer treatment is the forced differentiation of cancer cells . Here , we show that the treatment of the novel small molecule , CG500354 , into CD133-expressing human primary GBM cells induces growth arrest by cell cycle regulators , p53 , P38936 , p27 and phase-specific cyclins , and neural differentiation , as confirmed by neural progenitor/precursor markers , nestin , P14136 and Tuj1 . When GBM-derived cells caused the tumors in NOD/SCID mice , CG500354 induced GBM-derived cells differentiation into Tuj1 and P14136 expressing cells . We next demonstrated that CG500354 plays a tumor-suppressive role via DB02527 /CREB signaling pathway . CG500354 increases not only the extracellular DB02527 level but also the protein level of PKA and CREB . Additionally , both mimetic substances , DB02587 and DB01954 , revealed comparable results with CG500354 . Our findings indicate that induction of growth arrest and neural differentiation via DB02527 /CREB signaling pathway by CG500354 treatment suggests the novel targeting of Q08499 in the development of new drugs for brain tumor therapy . 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 . DB00952 has a selective inhibitory effect on trigeminovascular neurones at central P08908 and 5-HT(1B/1D) receptors in the cat : implications for migraine therapy . The triptans are agonists at serotonin (5-HT)1B/1D receptors ; however , they are also active at P08908 and P30939 receptors . We conducted this series of experiments to further elucidate the site of action of naratriptan using a well-established animal model of trigeminovascular stimulation . Following electrical stimulation of the superior sagittal sinus of the cat , single cell responses ( n=83 ) were recorded in the trigeminal nucleus caudalis . Most cells ( 91 % ) also responded to electrical and mechanical stimulation of cutaneous or mucosal facial receptive fields . The microiontophoretic application of naratriptan resulted in a significant suppression of the response to sagittal sinus stimulation ( response suppressed by 47 +/- 4 % , P < 0.001 ) . The effect of naratriptan was significantly attenuated by application of either the 5-HT(1B/1D) receptor antagonist GR-127935 ( P < 0.001 ) or the P08908 antagonist WAY-100635 ( P < 0.05 ) . The response of single cells to receptive field stimulation was also suppressed by microiontophoretic application of naratriptan , but by only 20 +/- 3 % . Intravenous administration of naratriptan resulted in a similar selective suppression of sagittal sinus vs. receptive field responses in trigeminal neurones . These results indicate that naratriptan has a central effect in the trigeminovascular system , selectively inhibiting afferent activity in craniovascular neurones , via both 5-HT(1B/1D) and P08908 receptors . DB06016 ( RGH-188 ) , a potent D3/D2 dopamine receptor partial agonist , binds to dopamine D3 receptors in vivo and shows antipsychotic-like and procognitive effects in rodents . We investigated the in vivo effects of orally administered cariprazine ( RGH-188 ; trans-N-{4-[2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-ethyl]-cyclohexyl}-N',N'-dimethyl-urea ) , a D(3)/ P14416 partial agonist with ∼10-fold preference for the D(3) receptor . Oral bioavailability of cariprazine at a dose of 1mg/kg in rats was 52 % with peak plasma concentrations of 91ng/mL . DB06016 10mg/kg had good blood-brain barrier penetration , with a brain/plasma AUC ratio of 7.6:1 . In rats , cariprazine showed dose-dependent in vivo displacement of [ (3)H ] (+)-PHNO , a dopamine D(3) receptor-preferring radiotracer , in the D(3) receptor-rich region of cerebellar lobules 9 and 10 . Its potent inhibition of apomorphine-induced climbing in mice ( ED(50)=0.27mg/kg ) was sustained for 8h . DB06016 blocked amphetamine-induced hyperactivity ( ED(50)=0.12mg/kg ) and conditioned avoidance response ( CAR ) ( ED(50)=0.84mg/kg ) in rats , and inhibited the locomotor-stimulating effects of the noncompetitive DB01221 antagonists MK-801 ( ED(50)=0.049mg/kg ) and phencyclidine ( ED(50)=0.09mg/kg ) in mice and rats , respectively . It reduced novelty-induced motor activity of mice ( ED(50)=0.11mg/kg ) and rats ( ED(50)=0.18mg/kg ) with a maximal effect of 70 % in both species . DB06016 produced no catalepsy in rats at up to 100-fold dose of its CAR inhibitory ED(50) value . DB06016 0.02-0.08mg/kg significantly improved the learning performance of scopolamine-treated rats in a water-labyrinth learning paradigm . Though risperidone , olanzapine , and aripiprazole showed antipsychotic-like activity in many of these assays , they were less active against phencyclidine and more cataleptogenic than cariprazine , and had no significant effect in the learning task . The distinct in vivo profile of cariprazine may be due to its higher affinity and in vivo binding to D(3) receptors versus currently marketed typical and atypical antipsychotics . Expression of variant dihydrofolate reductase with decreased binding affinity to antifolates in Q14C87 -3 human leukemia cell lines resistant to trimetrexate . Various alterations of the dihydrofolate reductase ( P00374 ) gene are involved in resistance . In order to understand the mechanism that induce such gene alterations in human leukemia cells , we studied the expression products of P00374 gene in trimetrexate ( DB01157 ) - and/or methotrexate ( MTX ) -resistant sublines derived from a Q14C87 -3 human leukemia cell line . A 200-fold DB01157 -resistant subline ( Q14C87 -3/TMQ200 ) expressed the mutated P00374 mRNA , with a base change ( T --> C ) at the second position of codon 31 , as well as the wild type gene . A MTX-resistant subline derived from Q14C87 -3/TMQ200 ( Q14C87 -3/TMQ200-MTX500 ) showed a further increase in the expression of the mutated P00374 mRNA , compared to Q14C87 -3/TMQ200 , with a marked decrease of expression of the wild type P00374 mRNA , which is confirmation of amplification of the mutated P00374 gene . By contrast , a 10,000-fold MTX-resistant subline ( Q14C87 -3/MTX10,000 ) over-expressed the wild type P00374 mRNA , which is confirmation of amplification of the wild type gene . Increased levels of the P00374 enzyme in these sublines were proportional to expression levels of the P00374 mRNA . The P00374 enzyme expressed in Q14C87 -3/TMQ200-MTX500 cells showed a 40-fold increase in the Ki values for both MTX and DB01157 , compared with values for the wild type P00374 expressed in both Q14C87 -3/MTX10,000 and its parent cell line . These findings suggest that the altered P00374 gene , which was introduced in Q14C87 -3 cells by exposure to DB01157 , gave rise to a variant enzyme with reduced affinity to antifolates , and that complex P00374 alterations confer drug-resistant phenotypes in antifolate-resistance . Structural difference between the antifolates could be important in the introduction of the differential P00374 gene alterations in the antifolate resistance . Immunohistochemical detection of components of the insulin-like growth factor system during skeletal muscle growth in the pig . The insulin-like growth factor ( IGF ) system plays an important role in postnatal somatic and skeletal muscle growth in pigs . There is little information on the occurrence and distribution of components of the IGF system in postnatal porcine skeletal muscle . P05019 , IGF receptor 1 ( P08069 ) and the IGF-binding proteins P08833 and -3 in longissimus dorsi and triceps brachii were localized in muscle biopsies from 12 commercially crossbred pigs aged from 28 to 199 days as well as from the sire generation , by immunohistochemistry . Plasma P05019 concentrations were also determined using radio-immunoassays . Unlike other species , P05019 was localized in porcine skeletal muscle fibres . Staining intensity correlated with the highest plasma P05019 levels and phases of intensive muscle growth from the 11th to 22nd week . The pattern of P08069 immunostaining , which was strong , correlated with that of P05019 , P08069 was also localized in endomysial tissues . P08833 was not detected within muscle fibres , but was found in the endomysium and vessel walls , while P17936 was localized with DB01277 and its receptor . Higher magnification revealed that P08069 , P17936 and probably P05019 appeared in the tubular system . Inhibitory as well as stimulating controls of P08833 and -3 on IGF functions are discussed , which may maintain a balance between autocrine growth promoting activities of P05019 and P08069 . A new model of cell cycle-regulated transcription : repression of the cyclin A promoter by P05231 -1 and anti-repression by E2F . Cell cycle regulation of the cyclin A gene is determined by a bipartite repressor binding site in the region of the basal promoter , termed CDE-CHR , which also controls the expression of cell cycle genes upregulated in S or G2 ( such as cdc25C ) . The CDE-CHR in the cyclin A promoter is recognized by both E2F complexes and P05231 -1 , but the contribution of each of these factors in cell cycle regulation is unknown . In the present study , we have introduced mutations into the cyclin A promoter which lead to either a loss or enhancement of E2F binding , while having only marginal effects on the interaction with P05231 -1 . Unlike mutants deficient for P05231 -1 binding , promoter variants lacking E2F binding showed an unchanged repression in G0 , thus identifying P05231 -1 as the principal repressor of the cyclin A gene . The same mutants did show , however , a delayed derepression while a mutation leading to increased E2F binding resulted in premature up-regulation . These findings clearly suggest that E2F contributes to the correct timing of cyclin A transcription , presumably by acting as an anti-repressor . In agreement with this conclusion , we find that the cyclin A promoter only poorly interacts with Q16254 , which is the major E2F family member in G0 cells , while a clear binding is seen with Q01094 and -3 , which are up-regulated in late P55008 . Estrogen upregulates endothelial nitric oxide synthase gene expression in fetal pulmonary artery endothelium . NO , produced by endothelial NO synthase ( P29474 ) , is a key mediator of pulmonary vasodilation during cardiopulmonary transition at birth . The capacity for NO production is maximal at term because pulmonary P29474 expression increases during late gestation . Since fetal estrogen levels rise markedly during late gestation and there is indirect evidence that the hormone enhances nonpulmonary NO production in adults , estrogen may upregulate P29474 in fetal pulmonary artery endothelium . Therefore , we studied the direct effects of estrogen on P29474 expression in ovine fetal pulmonary artery endothelial cells ( PAECs ) . DB00783 caused a 2.5-fold increase in NOS enzymatic activity in PAEC lysates . This effect was evident after 48 hours , and it occurred in response to physiological concentrations of the hormone ( 10(-10) to 10(-6) mol/L ) . The increase in NOS activity was related to an upregulation in P29474 protein expression , and P29474 mRNA abundance was also enhanced . P03372 antagonism with DB00947 completely inhibited estrogen-mediated P29474 upregulation , indicating that estrogen receptor activation is necessary for this response . In addition , immunocytochemistry revealed that fetal PAECs express estrogen receptor protein . Furthermore , transient transfection assays with a specific estrogen-responsive reporter system have demonstrated that the endothelial estrogen receptor is capable of estrogen-induced transcriptional transactivation . Thus , estrogen upregulates P29474 gene expression in fetal PAECs through the activation of PAEC estrogen receptors . This mechanism may be responsible for pulmonary P29474 upregulation during late gestation , thereby optimizing the capacity for NO-mediated pulmonary vasodilation at birth . DB00644 as a cell proliferation regulator : mechanism of action and evolutionary implications . DB00644 ( DB00644 ) is well known as the central regulator of the reproductive system through its stimulation of gonadotropin release from the pituitary . Studies on DB00644 have demonstrated that DB00644 has both stimulatory and inhibitory effects on cell proliferation depending on the cell type ; however , the mechanisms of these effects remain to be elucidated . Against this background we used four human cell lines , TSU-Pr1 , Jurkat , HHUA and DU145 , and newly found that DB00644 increased or decreased the colony-formation depending on the cell line . Moreover , we demonstrated that the stimulatory and inhibitory effects of DB00644 exhibit distinct ligand selectivities . In order to investigate the molecular basis of these phenomena , analyses of the expression of human DB00644 receptors were performed and , moreover , the effects of DB00644 were analyzed under conditions in which human DB00644 receptors were knocked down by the technique of RNA interference . Consequently , it was found that human type II P30968 , which had been suspected of being nonfunctional because of alterations in its sequence , is involved in the effects of DB00644 on cell proliferation . In this article , the influence of the autocrine activities of the cells is also reviewed , focusing on the characteristics of substances secreted from the four cell lines . Based on recent studies of DB00644 and its receptors and our up-to-date findings , the evolutionary implications of DB00644 action are discussed . Role of phosphodiesterases in the regulation of gonadotropin- releasing hormone secretion in GT1 cells . Increases in the level of DB02527 stimulate the secretion of DB00644 from GT1 DB00644 neuronal cells . We hypothesized that cyclic nucleotide phosphodiesterases ( PDEs ) , the enzymes that hydrolyze DB02527 , may constitute a negative feedback signaling mechanism for DB00644 regulation by decreasing the level of DB02527 . GT1 cells were shown to express three PDEs by RT-PCR analysis : the DB02527 -specific Q07343 and Q08499 and the calmodulin-dependent Q01064 . A splice variant of Q08499 , PDE4D3 , which is activated when phosphorylated by DB02527 -dependent protein kinase ( PKA ) , was identified in GT1 cells by Western analysis . Consistent with PDEs negatively regulating DB00644 secretion , treatment with the nonselective PDE inhibitor , DB07954 , stimulated DB00644 secretion 137 % in 30-min static cultures . Furthermore , treatment with the DB05876 -specific inhibitors DB01954 and RS-25344 increased DB00644 secretion 48 and 125 % , while treatment with the PDE1-specific inhibitor 8-MeoM- DB07954 only caused a modest increase of 28 % . In perifusion studies a rapid multi-fold stimulation of DB00644 secretion was observed following treatment with DB07954 , DB01954 or RS-25344 . In conclusion , the level of PDE activity appears to be an important negative feedback signal for DB00644 secretion . We hypothesize that activation of PDE4D3 by PKA may constitute a negative feedback signaling pathway which participates in the regulation of DB02527 levels . Systems biology of autosomal dominant polycystic kidney disease ( ADPKD ) : computational identification of gene expression pathways and integrated regulatory networks . To elucidate the molecular pathways that modulate renal cyst growth in ADPKD , we performed global gene profiling on cysts of different size ( < 1 ml , n = 5 ; 10-20 ml , n = 5 ; > 50 ml , n = 3 ) and minimally cystic tissue ( Q8IVS2 , n = 5 ) from five PKD1 human polycystic kidneys using Affymetrix HG-U133 Plus 2.0 arrays . We used gene set enrichment analysis to identify overrepresented signaling pathways and key transcription factors ( TFs ) between cysts and Q8IVS2 . We found down-regulation of kidney epithelial restricted genes ( e.g. nephron segment-specific markers and cilia-associated cystic genes such as P35680 , P08F94 , Q13099 and Q717R9 ) in the renal cysts . On the other hand , PKD1 cysts displayed a rich profile of gene sets associated with renal development , mitogen-mediated proliferation , cell cycle progression , epithelial-mesenchymal transition , hypoxia , aging and immune/inflammatory responses . Notably , our data suggest that up-regulation of Wnt/beta-catenin , pleiotropic growth factor/receptor tyrosine kinase ( e.g. IGF/ P08069 , FGF/FGFR , P01133 / P00533 , P15692 /VEGFR ) , G-protein-coupled receptor ( e.g. PTGER2 ) signaling was associated with renal cystic growth . By integrating these pathways with a number of dysregulated networks of TFs ( e.g. P11831 , MYC , Q01094 , P16220 , Q9UJU2 , P36402 , P35680 / P20823 and P41235 ) , our data suggest that epithelial dedifferentiation accompanied by aberrant activation and cross-talk of specific signaling pathways may be required for PKD1 cyst growth and disease progression . Pharmacological modulation of some of these signaling pathways may provide a potential therapeutic strategy for ADPKD . RILM : a web-based resource to aid comparative and functional analysis of the insulin and DB01277 receptor family . The metazoan receptors for insulin ( P06213 ) , insulin-like growth factor 1 ( P08069 ) , and other insulin-like molecules are transmembrane tyrosine kinases involved in the regulation of cell size , cell proliferation , development , signaling of nutritional and environmental conditions , and aging . Historically , mutations in the human insulin receptor have been studied because such changes often lead to severe insulin resistance . More recently , amino acid sequence alterations in the insulin receptor-like receptors of Drosophila melanogaster and Caenorhabditis elegans , as well as in the mouse insulin receptor have been the focus of attention . These modifications can have profound effects on growth , body size , metabolism , and aging . To integrate the many findings on insulin/IGF1 receptor structure and function across species we have created " Receptors for P01308 and P01308 -like Molecules " ( RILM ) , a curated computer-based resource that displays residue-by-residue information on sequence homology , three-dimensional structure , structure/function annotation , and documented mutations . The resource includes data obtained from sequence and structure analysis tools , primary database resources , and published reports . The information is integrated via a structure-based multiple sequence alignment of diverse members of the family . RILM was designed to provide easy access to multiple data types that could prove useful in the analysis of the effect of mutations on protein structure and ligand binding within this receptor family . RILM is available at www.biochem.ucl.ac.uk/RILM . DB06366 : evolving therapeutic strategies in the management of P04626 -overexpressing breast cancer . INTRODUCTION : P04626 overexpression or amplification is present in approximately one-fifth of breast cancers and historically was associated with aggressive disease and poorer prognosis . The introduction of the humanized monoclonal antibody trastuzumab dramatically improved disease-free survival ( DFS ) and overall survival ( OS ) in this subgroup . As the majority of patients with metastatic disease ultimately develop resistance to trastuzumab , a need exists for more effective targeted therapies . DB06366 is an anti- P04626 /neu-targeted therapy in the late stages of clinical development . The combination of pertuzumab , trastuzumab and docetaxel has been found to have an OS benefit in patients with P04626 positive metastatic breast cancer ( MBC ) when used in the first-line setting . This reflects a new standard of care , and pertuzumab was recently approved for this indication by the Food and Drug Administration ( FDA ) . The efficacy of pertuzumab and trastuzumab in conjunction with chemotherapy is currently being evaluated in the adjuvant setting . AREAS COVERED : This article provides an overview of preclinical investigations in addition to reviewing pertinent Phase I , Phase II and Phase III clinical trials . EXPERT OPINION : DB06366 , in combination with the humanized monoclonal antibody trastuzumab , and docetaxel is a standard of care for patients with previously untreated metastatic breast cancer based on the CLEOPATRA study showing a survival benefit . There is no increase in cardiac toxicity with the combined P04626 -targeted therapy . Future issues will address appropriate sequencing and combination with other anti- P04626 -targeted therapies and/or chemotherapy . Binding of gonadotrophin releasing hormone ( DB00644 ) by bovine anterior pituitary plasma membranes and purified P30968 protein ( DB00644 .R ) . [ 125I ] -( DB00644 ) concentration dependence binding curves using isolated bovine anterior pituitary plasma membranes , intact and solubilized , and purified P30968 protein , are compared . In all instances the concentration dependence binding curves had a stepwise character here interpreted as multisigmoid , with several steep increases and plateaus . These curves are compatible with the existence of several binding sites for DB00644 . Purification of the P30968 protein ( DB00644 .R ) resulted in about 500 000-fold increase of binding activity and yielded a single protein species on polyacrylamide gel electrophoresis in SDS , of estimated molecular weight 60 000 . Similarity of DB00644 binding by the purified protein with that of intact and solubilized plasma membranes suggested that a single protein was responsible for the binding in each instance . Thus heterogeneity of DB00644 binding is likely attributable to phase transitions of a single receptor protein into different allosteric forms . The data suggest that positive cooperativity is involved in the studied system . [ 5-hydroxytryptamine ( serotonin ) receptors -- nomenclature and classification of types and subtypes ] . 5-HT receptors represent a superfamily of receptors with the largest known number of receptor subtypes . At present 15 receptor subtypes of three groups has been recognized . The 5-HT1 subfamily of receptors contains subtypes P08908 , P28222 , P28221 , P28566 , and P30939 ; activation of all of them results in the inhibition of adenylylcyclase . The subfamily of 5-HT2 contains subtypes 5- Q13049 , P41595 , and P28335 ; their activation leads to the stimulation of P98160 . Finally , subfamily of miscellaneous 5-HT receptors contains subtypes 5- Q9H205 , Q13639 , 5-HT5 , P50406 , and P34969 ; some of them has been cloned , however , our knowledge on their function is still minimal . 5-HT receptors participate in many physiological functions and a disturbance in serotonergic neurotransmission might cause several types of disease . 5-HT plays an important role in depression ; to cure this disease , drugs which increase levels of this neurotransmitter are used . A new drug group called Selective Serotonin Reuptake Inhibitors ( SSRI ) has been recently discovered . These drugs block the reuptake of 5-HT into nerve endings . There is an intensive search for new selective agonists as well as antagonists which could be use not only in the classification of receptor subtypes but which also possess certain therapeutic potential .

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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 . P36888 inhibitors : a story of the old and the new . PURPOSE OF REVIEW : Ever since the recognition that P07333 -like tyrosine kinase 3 ( P36888 ) mutations exert a profound negative prognostic impact on the clinical outcome of patients with acute myeloid leukemia ( AML ) , researchers have sought to find effective small-molecule inhibitors of this receptor tyrosine kinase . This review will attempt to provide a survey of the P36888 inhibitors currently under investigation and provide a discussion on their current status in clinical trials . RECENT FINDINGS : Over the past 10 years , a number of different compounds have been studied in vitro and clinically as P36888 inhibitors . The first inhibitors studied were hampered by cumbersome pharmacokinetics and a general lack of potency . However , some agents have shown promise in clinical trials with transient responses in AML . Newer compounds , such as DB05213 , have demonstrated profound selectivity and potency against the P36888 target , and are currently being investigated in clinical trials . SUMMARY : Clinical trials have so far demonstrated that inhibitors of P36888 do have clinical activity in patients with P36888 -mutant AML , although this activity is often transient and correlates with effective in-vivo suppression of the P36888 target . As newer , more potent agents are now entering advanced clinical trials , opportunities will emerge for real progress against this grim disease . A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors . P10721 or alpha-platelet-derived growth factor receptor ( alpha- P09619 ) activating mutations are the pathogenic mechanisms that characterize gastrointestinal stromal tumors ( GIST ) . Despite excellent responses to imatinib mesylate ( IM ) , patients are relapsing . We developed an IM-resistant GIST cell line ( GIST-R ) from the IM-sensitive GIST882 cell line ( GIST-S ) by growing these cells in IM . Gene expression profiling ( GEP ) of GIST-S , GIST-R cells and two IM resistant GIST patients demonstrated that P10721 is downregulated implying a major role in IM resistance . Instead , GIST-R cells have acquired IM resistance by overexpressing the oncogenic receptor tyrosine kinase - P30530 - in a ' kinase switch ' . Further , the two IM resistant GIST patients express P30530 and not c-Kit , seen by immunohistochemistry ( IHC ) . Real time reverse transcriptase-polymerase chain reaction and Western blotting of the GIST-S and GIST-R cells confirmed the switch from Kit to P30530 . In GIST-R , P30530 is tyrosine phosphorylated and its ligand growth-arrest-specific gene 6 is overexpressed implying autocrine activation . The kinase switch is associated with a morphological change from spindle to epithelioid . Molecular modeling of the kinase domain of mutant c-Kit ( V654A ) and P30530 showed no binding to IM but efficient binding to DB05216 , a novel c-Kit/ P30530 kinase inhibitor . DB05216 synergizes with docetaxel ( taxotere ) and is cytotoxic to GIST cells . 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 . In-vitro effects of the tyrosine kinase inhibitor imatinib on glioblastoma cell proliferation . Glioblastoma ( Q9BVC4 ) is the most malignant brain tumour in adults , causing the death of most patients within 9-12 months of diagnosis . Treatment is based on a combination of surgery , radiation therapy , and chemotherapy . With these treatment modalities , however , responses are extremely poor , so identification of novel treatment strategies is highly warranted . Platelet-derived growth factors ( PDGF ) and their receptors are commonly coexpressed in Q9BVC4 , suggesting that stimulation of autocrine PDGF receptors may contribute to their growth . Interest in these receptors as drug target for glioblastoma treatment has increased with the clinical availability of the P09619 kinase inhibitor antagonist imatinib mesylate ( STI571 ) . In this study , T98G and A172 human Q9BVC4 cell lines were analysed for their sensitivity to treatment with imatinib . In particular , we focussed our attention on analysis of DNA distribution by flow cytometry at different times of incubation with different imatinib concentrations ( 1-30 microM : ) . Our results show that imatinib induces growth arrest in T98G and A172 cells in the G(0)/G(1) phase of the cell cycle , at all the concentrations tested , as early as 24 h after treatment . However we have also seen , by means of annexin V staining , that at 20 and 30 microM : concentrations , in concomitance with a significant growth arrest in the G(0)/G(1) phase , there is an increase of apoptotic cells 48 h after treatment , suggesting that imatinib at low concentrations ( 1-10 microM : ) could act as a cytostatic agent whereas at high concentrations ( 20 , 30 microM : ) it mainly behaves as a cytotoxic agent . [ Structural analysis of abnormal hemoglobin by the polymerase chain reaction ( PCR ) of genomic DNA ] . Determination of the primary structure of abnormal Hbs on the basis of DNA sequencing of the globin gene obtained from a carrier of abnormal Hb was performed . DNA obtained from the leukocytes of the peripheral blood was amplified by the polymerase chain reaction ( PCR ) using the proper amplification primer set . Amplified DNA was digested with two different restriction endonucleases and cloned to vector M 13 mp 18 or mp 19 , which had been digested with the same enzymes . DNA sequencing was done by the dideoxy chain termination method using T 7 DNA polymerase , and the abnormal Hbs whose primary structure was determined were as follows : Hb Fukuoka [ beta 2 DB00117 (CAC/T) ---- DB00135 (TAT) ] , Hb Machida [ beta 6 DB00142 (GAG) ---- Gln ( CAG ) ] , Hb Hope [ beta 136 DB00145 ( P19440 ) ---- DB00128 ( Q6IB77 ) ] , Hb Hiroshima [ beta 146 DB00117 (CAC) ---- DB00128 (GAC) ] and Hb Kodaira [ beta 146 DB00117 (CAC) ---- Gln(CAA) ] . This method for determining the primary structure of abnormal Hbs might be more effective than the ordinary method , which involves amino acid analysis and amino acid sequencing of the abnormal peptide obtained from abnormal Hb . Enhanced histone deacetylase enzyme activity in primary myelofibrosis . We measured histone deacetylase ( HDAC ) activity in 17 patients with primary myelofibrosis ( PMF ) ; 19 with other myeloproliferative neoplasm ( Q9BQR3 ) and 16 normal volunteers . Significantly elevated HDAC levels were shown in patients with PMF compared with other Q9BQR3 patients and normal volunteers ( p < 0.05 ) . Sixteen patients with PMF were also studied for correlation between O60674 mutation status and HDAC levels ; no significant correlation was found . We further correlated HDAC levels with clinical features in PMF : there was no correlation with WBC , platelet counts , Hb levels or degree of bone marrow fibrosis , but HDAC levels were correlated to the degree of splenomegaly . This suggests that HDAC may be recruited as essential thrombocythemia or polycythemia vera progresses into myelofibrosis or PMF progresses into more advanced stage . We then used the qRT-PCR cycle threshold ( CT ) method to study which HDACs were elevated in PMF . The results showed that , in general , Class 1 HDACs were elevated ( Q13547 ,2,8 ) except O15379 , Class II HDACs were depressed ( P56524 ,5 ) except Q9UBN7 and 10 , and Class III HDACs were generally elevated . The current study may form the basis for using HDAC inhibitor in the treatment of patients with PMF and may implicate a possible role of HDAC in the association of pathogenesis of PMF . Guanylate cyclase C-mediated antinociceptive effects of linaclotide in rodent models of visceral pain . BACKGROUND DB08890 is a novel , orally administered investigational drug currently in clinical development for the treatment of constipation-predominant irritable bowel syndrome ( IBS-C ) and chronic idiopathic constipation . Visceral hyperalgesia is a major pathophysiological mechanism in IBS-C . Therefore , we investigated the anti-nociceptive properties of linaclotide in rodent models of inflammatory and non-inflammatory visceral pain and determined whether these pharmacological effects are linked to the activation of guanylate cyclase C ( P25092 ) . METHODS Orally administered linaclotide was evaluated in non-inflammatory acute partial restraint stress ( PRS ) and acute water avoidance stress ( P42768 ) models in Wistar rats , and in a trinitrobenzene sulfonic acid ( TNBS ) -induced inflammatory model in Wistar rats and P25092 null mice . KEY RESULTS In TNBS-induced colonic allodynia , linaclotide significantly decreased the number of abdominal contractions in response to colorectal distension without affecting the colonic wall elasticity change in response to distending pressures after TNBS . However , linaclotide had no effect on visceral sensitivity under basal conditions . In addition , linaclotide significantly decreased colonic hypersensitivity in the PRS and P42768 models . In wild type ( wt ) and P25092 null mice , the instillation of TNBS induced similar hyperalgesia and allodynia . However , in post-inflammatory conditions linaclotide significantly reduced hypersensitivity only in wt mice , but not in P25092 null mice . CONCLUSIONS & INFERENCES These findings indicate that linaclotide has potent anti-nociceptive effects in several mechanistically different rodent models of visceral hypersensitivity and that these pharmacological properties of linaclotide are exerted through the activation of the P25092 receptor . Therefore , linaclotide may be capable of decreasing abdominal pain in patients suffering from IBS-C . P15692 ₁₆₄ isoform specific regulation of T-cell-dependent experimental colitis in mice . BACKGROUND : Inflammatory bowel disease ( Q9UKU7 ) consists of Crohn 's disease ( CD ) and ulcerative colitis ( UC ) , two widespread diseases of unknown , multifactorial etiology . Colitis pathology involves a pathological angiogenic response where increases in vascular density participate in colitis histopathology . Vascular endothelial growth factor-A ( P15692 ) is a potent angiogenesis stimulator known to be involved in pathological angiogenesis in several diseases including colitis . However , the pathogenic importance of specific P15692 isoforms during T-cell-mediated experimental colitis remains largely unknown . METHODS : The P01730 ⁺ CD45RB(high) T-cell transfer model of experimental colitis was used for these studies . The P15692 lac-Z transgenic reporter mouse was used to examine specific cellular sources of P15692 production . The P15692 ₁₆₄ aptamer ( Macugen ) , adenoviral P15692 ₁₆₄ , and the DB08885 were used to evaluate pathological importance . RESULTS : P15692 lac-Z reporter mice experiments showed that both infiltrating T cells and local tissue cells produce P15692 in the colon during disease . Inhibition of P15692 ₁₆₄ using a highly selective RNA aptamer significantly attenuated P01730 ⁺ CD45RB(high) T-cell-dependent experimental colitis by reducing pathological angiogenesis and inflammatory pathology . Conversely , broad-spectrum P15692 inhibition with DB08885 did not attenuate disease , nor did adenoviral P15692 ₁₆₄ overexpression significantly alter colitis pathology . CONCLUSIONS : P15692 ₁₆₄ is actively produced by multiple cell types during T-cell-mediated colitis . Importantly , specific inhibition of the P15692 ₁₆₄ isoform during T-cell-mediated colitis dose-dependently attenuated disease progression , while broad-scale inhibition of all P15692 isoforms was not therapeutically beneficial . 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 . DB00898 dose-dependently elicits opposing regulatory effects on hippocampal AMPA receptor P42262 subunits through a zeta inhibitory peptide-sensitive kinase in adolescent and adult Sprague-Dawley rats . AMPA receptor P42262 subunits are strongly implicated in cognition , and prior work suggests that these subunits may be regulated by atypical protein kinase C ( aPKC ) isoforms . The present study assessed whether hippocampal and cortical AMPA receptor P42262 subunit regulation may be an underlying factor in known age-related differences to cognitive-impairing doses of ethanol , and if aPKC isoforms modulate such responses . Hippocampal AMPA receptor P42262 subunit , protein kinase Mζ ( PKMζ ) , and PKCι/λ expression were elevated during adolescence compared to adults . 1 h following a low-dose ( 1.0-g/kg ) ethanol exposure , hippocampal AMPA receptor P42262 subunit serine 880 phosphorylation was decreased in adolescents , but was increased in adults . Age-dependent changes in P42262 subunit phosphorylation were paralleled by alterations in aPKC isoforms , and zeta inhibitory peptide ( Q8N5A5 ) administration prevented ethanol-induced increases in both in adults . DB00898 -induced changes in P42262 subunit phosphorylation were associated with delayed regulation in synaptosomal P42262 subunit expression 24 h later . A higher ethanol dose ( 3.5-g/kg ) failed to elicit changes in most measures in the hippocampus at either age . Similar to the hippocampus , analysis of cerebral cortical tissue also revealed age-related declines . However , no demonstrable effects were found following a low-dose ethanol exposure at either age . High-dose ethanol exposure reduced adolescent P42262 subunit phosphorylation and aPKC isoform expression that were again accompanied by delayed reductions in synaptosomal P42262 subunit expression . Together , these results suggest that P42262 -containing AMPA receptor modulation by aPKC isoforms is age- , region- and dose-dependently regulated , and may potentially be involved in developmentally regulated ethanol-induced cognitive impairment and other ethanol behaviors . 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 . Distinct mechanistic activity profile of pralatrexate in comparison to other antifolates in in vitro and in vivo models of human cancers . PURPOSE : This study evaluated mechanistic differences of pralatrexate , methotrexate , and pemetrexed . METHODS : Inhibition of dihydrofolate reductase ( P00374 ) was quantified using recombinant human P00374 . Cellular uptake and folylpolyglutamate synthetase ( Q05932 ) activity were determined using radiolabeled pralatrexate , methotrexate , and pemetrexed in NCI-H460 non-small cell lung cancer ( NSCLC ) cells . The tumor growth inhibition ( TGI ) was assessed using MV522 and NCI-H460 human NSCLC xenografts . RESULTS : Apparent K ( i ) values for P00374 inhibition were 45 , 26 , and > 200 nM for pralatrexate , methotrexate , and pemetrexed , respectively . A significantly greater percentage of radiolabeled pralatrexate entered the cells and was polyglutamylatated relative to methotrexate or pemetrexed . In vivo , pralatrexate showed superior anti-tumor activity in both NSCLC models , with more effective dose-dependent TGI in the more rapidly growing NCI-H460 xenografts . CONCLUSIONS : DB06813 demonstrated a distinct mechanistic and anti-tumor activity profile relative to methotrexate and pemetrexed . DB06813 exhibited enhanced cellular uptake and increased polyglutamylation , which correlated with increased TGI in NSCLC xenograft models . [ 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 . 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 . 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 . P23560 activation of P62158 -kinase kinase via transient receptor potential canonical channels induces the translation and synaptic incorporation of P42261 -containing calcium-permeable AMPA receptors . Glutamatergic synapses in early postnatal development transiently express calcium-permeable AMPA receptors ( CP-AMPARs ) . Although these P42262 -lacking receptors are essential and are elevated in response to brain-derived neurotrophic factor ( P23560 ) , little is known regarding molecular mechanisms that govern their expression and synaptic insertion . Here we show that P23560 -induced P42261 translation in rat primary hippocampal neurons requires the activation of mammalian target of rapamycin ( P42345 ) via calcium calmodulin-dependent protein kinase kinase ( CaMKK ) . Specifically , P23560 -mediated phosphorylation of threonine 308 ( T308 ) in AKT , a known substrate of CaMKK and an upstream activator of P42345 -dependent translation , was prevented by ( 1 ) pharmacological inhibition of CaMKK with STO-609 , ( 2 ) overexpression of a dominant-negative CaMKK , or ( 3 ) short hairpin-mediated knockdown of CaMKK . P42261 surface expression induced by P23560 , as assessed by immunocytochemistry using an extracellular N-terminal P42261 antibody or by surface biotinylation , was impaired following knockdown of CaMKK or treatment with STO-609 . Activation of CaMKK by P23560 requires transient receptor potential canonical ( TRPC ) channels as SKF-96365 , but not the DB01221 receptor antagonist d-APV , prevented P23560 -induced P42261 surface expression as well as phosphorylation of CaMKI , AKT(T308) , and P42345 . Using siRNA we confirmed the involvement of Q9UL62 and Q9Y210 subunits in P23560 -induced AKT(T308) phosphorylation . The P23560 -induced increase in mEPSC was blocked by IEM-1460 , a selected antagonist of CP-AMPARs , as well as by the specific repression of acute P42261 translation via siRNA to P42261 but not P42262 . Together these data support the conclusion that newly synthesized P42261 subunits , induced by P23560 , are readily incorporated into synapses where they enhance the expression of CP-AMPARs and synaptic strength . 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 . Characterization of rat rostral raphe primary cultures : multiplex quantification of serotonergic markers . Previous reports establishing raphe cultures typically yield less than 1 % serotonin ( 5-HT ) -positive neurons and are impractical for transcriptional studies . In this study , we have established primary cultures enriched in 5-HT neurons and quantified the proportion of cells expressing serotonergic and non-serotonergic markers . We have also shown the feasibility of using the multiplex real-time PCR technique to measure the relative amounts of RNA for some of these markers . Rostral raphe cells derived from E13-15 rat embryos were cultured for 7 days and analyzed by quantitative immunofluorescence and western blot analysis . In these cultures , approximately 8 % of neurons were immunopositive for serotonergic markers ( 5-HT or tryptophan hydroxylase ( P17752 ) ) . The percentage of cells labeled for P14136 ( glial marker ) , tyrosine hydroxylase ( catecholaminergic ) , and Q05329 /67 ( GABAergic ) was 5 , 1 , and 54 % , respectively . Transcription factors Q13127 / Q13127 and Deaf-1 were present in 9 and 98 % of cells , respectively . Multiplex quantitative RT-PCR ( Q-PCR ) analysis was done for Q8IWU9 , P08908 receptor or Deaf-1 RNAs paired with P04406 RNA as control . Using this approach , standard curves for each RNA were obtained over 200-fold concentration range of dilution with r2 values > 0.99 . The relative abundances determined by Q-PCR are consistent with the expression of Q8IWU9 > Deaf-1 > P08908 receptor RNA in serotonergic raphe cells . The standard error of Q8IWU9 RNA levels between cultures was < 20 % , indicating a consistent purity of 5-HT neurons . Thus , we have generated a highly consistent and reproducible model system that is enriched in 5-HT neurons and that will be valuable in future investigation of serotonergic regulation . 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 . Targeting PIM kinases impairs survival of hematopoietic cells transformed by kinase inhibitor-sensitive and kinase inhibitor-resistant forms of P36888 and P11274 / P00519 . Previous studies have shown that activation of the signal transducer and activator of transcription 5 ( P42229 ) plays an essential role in leukemogenesis mediated through constitutive activated protein tyrosine kinases ( PTK ) . Because PIM-1 is a P42229 target gene , we analyzed the role of the family of PIM serine/threonine kinases ( PIM-1 to PIM-3 ) in PTK-mediated transformation of hematopoietic cells . Ba/ P13726 cells transformed to growth factor independence by various oncogenic PTKs ( P41212 / O60674 , P41212 / Q16288 , P41212 / P00519 , P11274 / P00519 , P36888 -ITD , and H4/PDGFbetaR ) show abundant expression of PIM-1 and PIM-2 . Suppression of PIM-1 activity had a negligible effect on transformation . In contrast , expression of kinase-dead PIM-2 mutant ( PIM-2KD ) led to a rapid decline of survival in Ba/ P13726 cells transformed by P36888 -ITD but not by other oncogenic PTKs tested . Coexpression of PIM-1KD and PIM-2KD abrogated growth factor-independent growth of Ba/ P13726 transformed by several PTKs , including P11274 / P00519 . Targeted down-regulation of PIM-2 by RNA interference ( RNAi ) selectively abrogated survival of Ba/ P13726 cells transformed by various P36888 ( P36888 ) -activating mutants [ internal tandem duplication ( ITD ) and kinase domain ] and attenuated growth of human cell lines containing P36888 mutations . Interestingly , cells transformed by P36888 and P11274 / P00519 mutations that confer resistance to small-molecule tyrosine kinase inhibitors were still sensitive to knockdown of PIM-2 , or PIM-1 and PIM-2 by RNAi . Our observations indicate that combined inactivation of PIM-1 and PIM-2 interferes with oncogenic PTKs and suggest that PIMs are alternative therapeutic targets in PTK-mediated leukemia . Targeting the PIM kinase family could provide a new avenue to overcome resistance against small-molecule tyrosine kinase inhibitors . 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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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 . DB00480 inhibits osteoclastogenesis , survival factors and bone-remodeling markers in multiple myeloma . Osteolytic bone disease in multiple myeloma ( MM ) is caused by enhanced osteoclast ( OCL ) activation and inhibition of osteoblast function . DB00480 and bortezomib have shown promising response rates in relapsed and newly diagnosed MM , and bortezomib has recently been reported to inhibit OCLs . We here investigated the effect of lenalidomide on OCL formation and osteoclastogenesis in comparison with bortezomib . Both drugs decreased alpha V beta 3-integrin , tartrate-resistant acid phosphatase-positive cells and bone resorption on dentin disks . In addition , both agents decreased receptor activator of nuclear factor-kappaB ligand ( O14788 ) secretion of bone marrow stromal cells ( BMSCs ) derived from MM patients . We identified PU.1 and pERK as major targets of lenalidomide , and nuclear factor of activated T cells of bortezomib , resulting in inhibition of osteoclastogenesis . Furthermore , downregulation of cathepsin K , essential for resorption of the bone collagen matrix , was observed . We demonstrated a significant decrease of growth and survival factors including macrophage inflammatory protein-alpha , B-cell activating factor and a proliferation-inducing ligand . Importantly , in serum from MM patients treated with lenalidomide , the essential bone-remodeling factor O14788 , as well as the O14788 / O00300 ratio , were significantly reduced , whereas osteoprotegerin ( O00300 ) was increased . We conclude that both agents specifically target key factors in osteoclastogenesis , and could directly affect the MM-OCL-BMSCs activation loop in osteolytic bone disease . P50750 regulates AR promoter selectivity and cell growth through serine 81 phosphorylation . Previously we determined that S81 is the highest stoichiometric phosphorylation on the androgen receptor ( AR ) in response to hormone . To explore the role of this phosphorylation on growth , we stably expressed wild-type and S81A mutant AR in LHS and LAPC4 cells . The cells with increased wild-type AR expression grow faster compared with parental cells and S81A mutant-expressing cells , indicating that loss of S81 phosphorylation limits cell growth . To explore how S81 regulates cell growth , we tested whether S81 phosphorylation regulates AR transcriptional activity . LHS cells stably expressing wild-type and S81A mutant AR showed differences in the regulation of endogenous AR target genes , suggesting that S81 phosphorylation regulates promoter selectivity . We next sought to identify the S81 kinase using ion trap mass spectrometry to analyze AR-associated proteins in immunoprecipitates from cells . We observed cyclin-dependent kinase (CDK)9 association with the AR . P50750 phosphorylates the AR on S81 in vitro . Phosphorylation is specific to S81 because P50750 did not phosphorylate the AR on other serine phosphorylation sites . Overexpression of P50750 with its cognate cyclin , P12004 T , increased S81 phosphorylation levels in cells . Small interfering RNA knockdown of P50750 protein levels decreased hormone-induced S81 phosphorylation . Additionally , treatment of LNCaP cells with the P50750 inhibitors , 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole and DB03496 , reduced S81 phosphorylation further , suggesting that P50750 regulates S81 phosphorylation . Pharmacological inhibition of P50750 also resulted in decreased AR transcription in LNCaP cells . Collectively these results suggest that P50750 phosphorylation of AR S81 is an important step in regulating AR transcriptional activity and prostate cancer cell growth . 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 . 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 . 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 . 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 . 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 . The second generation of P35354 inhibitors : clinical pharmacological point of view . DB00580 , parecoxib , etoricoxib and lumiracoxib represent the second generation of selective P35354 inhibitors . In comparison to the first generation , they show an at least equivalent efficacy in the treatment of pain and inflammation . However , the postulated gain of safety is yet difficult to determine and seems to be , if any , small . Dexamethasone reverses adrenalectomy-induced neuronal de-differentiation in midbrain raphe-hippocampus axis . Differentiation leads to specific morphological and biochemical characteristics . We examined whether epigenetic factors ( e.g. , glucocorticoids ) are required to maintain neuronal differentiation in the adult brain . In the midbrain , adrenalectomy ( P10109 ) ( 1-2 wk ) reduced the size of tryptophan hydroxylase ( WH ) -immunoreactive ( IR ) neurons . P10109 rats exposed to short-term ( 24-72-h ) dexamethasone ( ST-DEX ) in the drinking saline ( 10 mg/l ) showed an increase in WH protein , somal area and dendritic size of WH-IR neurons . In the hippocampus , P10109 for 2-3 mo ( long-term ; LT ) reduced Nissl staining , calbindin ( DB09061 ) -IR and P08908 receptor mRNA in the granular cell layer , and the size of the molecular layer and its DB09061 -IR dendrites . Small vimentin ( Vim ) -IR glial cells emerged in the granular layer . ST-DEX after LT- P10109 rapidly induced a recovery of P08908 mRNA , Nissl labeling and DB09061 -IR in the granule cell layer . In the molecular layer , there was an increase in the area and in the number of DB09061 -IR dendrites . Furthermore , the Vim-IR glial cells were enlarged in size and branching . The rate of cell proliferation was studied in these animals . Immunostaining with antibodies against proliferating cell nuclear antigen ( P12004 ) and use of bromouridine argue against enhanced neurogenesis after ST-DEX in LT- P10109 . We propose that glucocorticoids induce and maintain differentiation of serotonergic and DB09061 -IR neurons in the midbrain-hippocampal axis . A neuronotrophic role for the glial P08908 receptor is suggested . 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 . Synaptically-competent neurons derived from canine embryonic stem cells by lineage selection with P01133 and Q13253 . Pluripotent stem cell lines have been generated in several domestic animal species ; however , these lines traditionally show poor self-renewal and differentiation . Using canine embryonic stem cell ( cESC ) lines previously shown to have sufficient self-renewal capacity and potency , we generated and compared canine neural stem cell ( cNSC ) lines derived by lineage selection with epidermal growth factor ( P01133 ) or Q13253 along the neural default differentiation pathway , or by directed differentiation with retinoic acid ( RA ) -induced floating sphere assay . Lineage selection produced large populations of P48431 + neural stem/progenitor cell populations and neuronal derivatives while directed differentiation produced few and improper neuronal derivatives . Primary canine neural lines were generated from fetal tissue and used as a positive control for differentiation and electrophysiology . Differentiation of P01133 - and Q13253 -directed cNSC lines in N2B27 with low-dose growth factors ( P23560 / P20783 or PDGFαα ) produced phenotypes equivalent to primary canine neural cells including 3CB2+ radial progenitors , Q8WUK0 + glia restricted precursors , P08670 +/ P14136 + astrocytes , and Q13509 +/ P11137 +/ P12036 +/ O15061 + neurons . Conversely , induction with RA and neuronal differentiation produced inadequate putative neurons for further study , even though appropriate neuronal gene expression profiles were observed by RT-PCR ( including P48681 , Q13509 , P78352 , Q16623 , Q8TBG9 , P11137 ) . Co-culture of cESC-derived neurons with primary canine fetal cells on canine astrocytes was used to test functional maturity of putative neurons . Canine ESC-derived neurons received functional GABA(A)- and AMPA-receptor mediated synaptic input , but only when co-cultured with primary neurons . This study presents established neural stem/progenitor cell populations and functional neural derivatives in the dog , providing the proof-of-concept required to translate stem cell transplantation strategies into a clinically relevant animal model . Transient transfection of GGH3-1 ' cells [ GH3 cells stably transfected with the gonadotropin-releasing hormone ( DB00644 ) receptor complementary deoxyribonucleic acid ] with the carboxyl-terminal of beta-adrenergic receptor kinase 1 blocks prolactin release : evidence for a role of the G protein beta gamma-subunit complex in DB00644 signal transduction . G proteins consist of heterotrimeric alpha- , beta- , and gamma-subunits . To assess the role of the beta gamma-subunit complex in P30968 -mediated signal transduction , GGH3-1 ' cells were transfected with plasmids PRK5-beta O14965 ( 495-689 ) containing complementary DNA ( cDNA ) of the carboxyl-terminal ( Gly495-Leu689 ) of beta-adrenergic receptor kinase 1 ( beta O14965 ) . GGH3-1 ' cells are GH3 cells that have been stably transfected with rat P30968 cDNA . The carboxyl region of beta O14965 ( Gly495-Leu689 ) binds to the beta gamma complex and thereby inhibits its action . Twenty-four hours after stimulation , PRL release , DB02527 release , and inositol phosphate ( IP ) production were measured in these cells and in control cells transfected with vector PRK5 cDNA alone . In cells expressing the beta O14965 -(495-689) sequence there was inhibition of basal PRL release ( 69.3 % ) , DB02527 release ( 61.2 % ) , and IP production ( 75.5 % ) compared to cells containing vector only . When cells expressing the beta O14965 fragment were stimulated with a DB00644 analog ( DB06719 ; 10(-7) M ) , maximal responses were inhibited ( 66.1 % for PRL release , 52.3 % for DB02527 release , and 79.1 % for IP production ) . Scatchard analysis of DB00644 analog binding was also performed in the two groups of transfected cells . No significant differences in Kd or receptor numbers were found between beta O14965 -(495-689)-transfected cells and control cells containing the vector alone . These data suggest a role for the beta gamma complex in mediation of DB02527 release , IP production , and hormone release in response to agonist occupancy of the P30968 . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . O14965 is not involved in Q9BZB8 phosphorylation and cyclin B1 mRNA polyadenylation during meiotic maturation of porcine oocytes . Regulation of mRNA translation by cytoplasmic polyadenylation is known to be important for oocyte maturation and further development . This process is generally controlled by phosphorylation of cytoplasmic polyadenylation element binding protein 1 ( Q9BZB8 ) . The aim of this study is to determine the role of O14965 in Q9BZB8 phosphorylation and the consequent Q9BZB8 -dependent polyadenylation of maternal mRNAs during mammalian oocyte meiosis . For this purpose , we specifically inhibited O14965 with DB05220 during meiotic maturation of porcine oocytes . Using poly(A)-test PCR method , we monitored the effect of O14965 inhibition on poly(A)-tail extension of long and short cyclin B1 encoding mRNAs as markers of Q9BZB8 -dependent cytoplasmic polyadenylation . Our results show that inhibition of O14965 activity impairs neither cyclin B1 mRNA polyadenylation nor its translation and that O14965 is unlikely to be involved in Q9BZB8 activating phosphorylation . 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 . 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 . Molecular mechanisms of patupilone resistance . DB03010 is an epothilone in advanced clinical development that has shown promising efficacy in heavily pretreated patients . This study aimed at characterizing the mechanisms of patupilone activity in resistant patients . To this end , we generated patupilone-resistant cells using two cellular models , the first characterized by high chemosensitivity and low class III beta-tubulin ( Q13509 ) expression ( A2780 ) , and the second by low chemosensitivity and high Q13509 expression ( OVCAR-3 ) . The obtained cell lines were named EPO3 and OVCAR-EPO , respectively . The same selection procedure was done in A2780 cells to generate a paclitaxel-resistant cell line ( TAX50 ) . Factors of resistance are expected to increase in the drug-resistant cell lines , whereas factors of drug sensitivity will be down-regulated . Using this approach , we found up-regulation of Q13509 in TAX50 , but not EPO3 , cells , showing that Q13509 mediates the resistance to paclitaxel but not to patupilone . Moreover , Q13509 was a factor of patupilone sensitivity because OVCAR-EPO cells exhibited a dramatic reduction of Q13509 and a concomitant sensitization to hypoxia and cisplatin-based chemotherapy . To identify the mechanisms underlying patupilone resistance , tubulin genes were sequenced , thereby revealing that a prominent mechanism of drug resistance is represented by point mutations in class I beta-tubulin . Overall , these results suggest that paclitaxel and patupilone have nonoverlapping mechanisms of resistance , thus allowing the use of patupilone for those patients relapsing after paclitaxel-based chemotherapy . Furthermore , patupilone represents a promising first-line option for the treatment of high-risk ovarian cancer patients , who exhibit high Q13509 levels and poor response to standard paclitaxel-platin chemotherapy . 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 . Serotonin and fluoxetine modulate bone cell function in vitro . Recent studies have proposed a role for serotonin and its transporter in regulation of bone cell function . In the present study , we examined the in vitro effects of serotonin and the serotonin transporter inhibitor fluoxetine " DB00472 " on osteoblasts and osteoclasts . Human mononuclear cells were differentiated into osteoclasts in the presence of serotonin or fluoxetine . Both compounds affected the total number of differentiated osteoclasts as well as bone resorption in a bell-shaped manner . RT-PCR on the human osteoclasts demonstrated several serotonin receptors , the serotonin transporter , and the rate-limiting enzyme in serotonin synthesis , tryptophan hydroxylase 1 ( Tph1 ) . Tph1 expression was also found in murine osteoblasts and osteoclasts , indicating an ability to produce serotonin . In murine pre-osteoclasts ( RAW264.7 ) , serotonin as well as fluoxetine affected proliferation and NFkappaB activity in a biphasic manner . Proliferation of human DB05914 ( O60682 ) and primary osteoblasts ( NHO ) , and 5- Q13049 receptor expression was enhanced by serotonin . DB00472 stimulated proliferation of O60682 and murine preosteoblasts ( MC3T3-E1 ) in nM concentrations , microM concentrations were inhibitory . The effect of fluoxetine seemed direct , probably through 5-HT2 receptors . Serotonin-induced proliferation of MC3T3-E1 cells was inhibited by the PKC inhibitor ( GF109203 ) and was also markedly reduced when antagonists of the serotonin receptors P41595 /C or 5- Q13049 /C were added . Serotonin increased osteoprotegerin ( O00300 ) and decreased receptor activator of NF-kappaB ligand ( O14788 ) secretion from osteoblasts , suggesting a role in osteoblast-induced inhibition of osteoclast differentiation , whereas fluoxetine had the opposite effect . This study further describes possible mechanisms by which serotonin and the serotonin transporter can affect bone cell function . 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 .

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interacts_with DB06212?

multiple_choice

The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . 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 . Autosomal-dominant hypophosphatemic rickets ( P30518 ) mutations stabilize Q9GZV9 . BACKGROUND : The gene for the renal phosphate wasting disorder autosomal-dominant hypophosphatemic rickets ( P30518 ) is Q9GZV9 , which encodes a secreted protein related to the fibroblast growth factors ( FGFs ) . We previously detected missense mutations R176Q , R179W , and R179Q in Q9GZV9 from P30518 kindreds . The mutations replace R residues within a subtilisin-like proprotein convertase ( Q969E3 ) cleavage site 176RHTR-179 ( RXXR motif ) . The goal of these studies was to determine if the P30518 mutations lead to protease resistance of Q9GZV9 . METHODS : The P30518 mutations were introduced into human Q9GZV9 cDNA clones with or without an N-terminal FLAG tag by site-directed mutagenesis and were transiently transfected into HEK293 cells . Protein expression was determined by Western analyses . RESULTS : Antibodies directed toward the C-terminal portion of Q9GZV9 revealed that the native Q9GZV9 protein resolved as 32 kD and 12 kD species in HEK293 conditioned media ; however , the three mutated proteins were detected only as the 32 kD band . An N-terminal FLAG-tagged native Q9GZV9 resolved as two bands of 36 kD and 26 kD when detected with a FLAG antibody , whereas the R176Q mutant resolved primarily as the 36 kD protein species . Cleavage of Q9GZV9 was not enhanced by extracellular incubation of Q9GZV9 with HEK293 cells . Native and mutant FGF-23s bound heparin . CONCLUSIONS : Q9GZV9 proteins containing the P30518 mutations are secreted , and produce polypeptides less sensitive to protease cleavage than wild-type Q9GZV9 . Therefore , the P30518 mutations may protect Q9GZV9 from proteolysis , thereby potentially elevating circulating concentrations of Q9GZV9 and leading to phosphate wasting in P30518 patients . [ Diagnosis and treatment of isolated methylmalonic acidemia ] . OBJECTIVE : To explore the clinical feature , therapeutic effect and prognosis of isolated methylmalonic acidemia . METHODS : The clinical characteristics , laboratory findings , treatment and outcome of 40 patients were retrospectively analyzed . The main treatment was a low-protein diet supplemented with L-carnitine and special milk free of leucine , valine , threonine and methionine . DB00115 was also given to cobalamin responders . The patients were followed up every 1-3 months . RESULTS : Mutations in the P22033 gene were identified in 30 of 33 patients who had accepted DNA testing . Thirty cases were treated and followed up regularly for from 1 month to 8 years . Eight cases had died , 8 had developed normal intelligence , among whom 4 from newborn screening were asymptomatic . Psychomotor developmental delay and mental retardation were present in 14 cases . The propionylcarnitine level , ratio of propionylcarnitine/acetylcarnitine in blood , methylmalonic acid and methylcitric acid levels in urine have decreased significantly , with the median values reduced respectively from 24.15 ( 7.92-81.02 ) μmol/L , 1.08 ( 0.38-6.01 ) , 705.34 ( 113.79-3078.60 ) and 7.71 ( 0.52-128.21 ) to 10.50 ( 3.00-30.92 ) μmol/L , 0.63 ( 0.25-2.89 ) , 166.23 ( 22.40-3322.21 ) and 3.96 ( 0.94-119.13 ) ( P < 0.05 ) . CONCLUSION : The prognosis of isolated methylmalonic acidemia may be predicted with the enzymatic subgroup , age at onset and cobalamin responsiveness . Outcome is unfavorable in neonatal patients and those who were non-responsive to cobalamin . Inhibition of Niemann-Pick-type C1-like1 by ezetimibe activates autophagy in human hepatocytes and reduces mutant α1-antitrypsin Z deposition . Autophagy can degrade aggregate-prone proteins , but excessive autophagy can have adverse effects . It would be beneficial if autophagy could be enhanced in a cell type-specific manner , but this has been difficult because the basic mechanism of autophagy is common . In the present study we found that inhibition of Niemann-Pick-type C1-like 1 ( Q9UHC9 ) by ezetimibe activates autophagy only in hepatocytes and small intestinal epithelia , but not in other cells . DB00973 induced accumulation of free cholesterol in the late endosome/lysosome and increased partitioning of a Ragulator component , Q6IAA8 , in rafts . The latter change led to down-regulation of mammalian target of rapamycin ( P42345 )C1 activity by decreasing P42345 recruitment to the late endosome/lysosome and activated autophagy . A primary effect of ezetimibe was found to be a decrease of free cholesterol in the plasma membrane , because all the results caused by ezetimibe were suppressed by supplementation of cholesterol as a methyl-β-cyclodextrin complex . By enhancing autophagy in human primary hepatocytes with ezetimibe , insoluble mutant α1-antitrypsin Z was reduced significantly . CONCLUSION : Inhibition of Q9UHC9 by ezetimibe activates autophagy in human hepatocytes by modulating cholesterol homeostasis . DB00973 may be used to ameliorate liver degeneration in α1-antitrypsin deficiency . The Drosophila insulin receptor activates multiple signaling pathways but requires insulin receptor substrate proteins for DNA synthesis . The Drosophila insulin receptor ( P30518 ) contains a 368-amino-acid COOH-terminal extension that contains several tyrosine phosphorylation sites in YXXM motifs . This extension is absent from the human insulin receptor but resembles a region in insulin receptor substrate ( P41252 ) proteins which binds to the phosphatidylinositol ( PI ) 3-kinase and mediates mitogenesis . The function of a chimeric P30518 containing the human insulin receptor binding domain ( hDIR ) was investigated in 32D cells , which contain few insulin receptors and no P41252 proteins . P01308 stimulated tyrosine autophosphorylation of the human insulin receptor and hDIR , and both receptors mediated tyrosine phosphorylation of Shc and activated mitogen-activated protein kinase . P35568 was required by the human insulin receptor to activate PI 3-kinase and p70s6k , whereas hDIR associated with PI 3-kinase and activated p70s6k without P35568 . However , both receptors required P35568 to mediate insulin-stimulated mitogenesis . These data demonstrate that the P30518 possesses additional signaling capabilities compared with its mammalian counterpart but still requires P35568 for the complete insulin response in mammalian cells . Q9GZV9 and matrix-metalloproteinases in patients with chronic kidney disease : are they associated with cardiovascular disease ? BACKGROUND : High cardiovascular risk in patients with chronic kidney disease ( CKD ) may be related to mineral disorder and microinflammation . Q9GZV9 ( Q9GZV9 ) is a phosphatonin and inhibitor of calcitriol synthesis , which is associated with poor prognosis in CKD patients starting dialysis . Matrix-metalloproteinases ( P08253 , P14780 ) contribute to myocardial remodeling and arterial calcification . Q9GZV9 and MMPs levels are altered in CKD , however , little is known about their association and relation to cardiovascular ( CV ) disease . METHODS : Standard laboratory parameters , plasma levels of P08253 , P14780 , Q9GZV9 , Q13219 and CV disease history were assessed in 80 patients with CKD 1-5 and 44 healthy control subjects . RESULTS : Q9GZV9 and P08253 ( assessed by ELISA ) were higher in CKD patients compared to controls . Q9GZV9 increased from CKD 3 , whereas P08253 increased only in CKD 5 . Q9GZV9 was positively associated with P08253 , adjusted to age , eGFR , phosphatemia , calcitriol and parathormone . Q9GZV9 independently correlated with parathormone and inversely with calcitriol , whereas P08253 was related to phosphatemia . Q9GZV9 was higher in subjects with a history of CV disease compared to those free of such history ( 559.0 vs.184.0 RU/ml ) , adjusted to age and eGFR . CONCLUSION : Our data suggest a possible relationship between Q9GZV9 , P08253 and CV disease in CKD . Potential causality of this association remains to be elucidated . Neuroprotection mediated via neurotrophic factors and induction of neurotrophic factors . Neurotrophins and other neurotrophic factors have been shown to support the survival and differentiation of many neuronal populations of the central and peripheral nervous system . Therefore , administering neurotrophic factors could represent an alternative strategy for the treatment of acute and chronic brain disorders . However , the delivery of neurotrophic factors to the brain is one of the largest obstacles in the development of effective therapy for neurodegenerative disorders , because these proteins are not able to cross the blood-brain barrier . The induction of growth factor synthesis in the brain tissue by systemically administered lipophilic drugs , such as beta-adrenoceptor agonists , shown to increase endogenous nerve growth factor ( P01138 ) synthesis in the brain , would be an elegant way to overcome these problems of application . Stimulation of beta-adrenoceptors with clenbuterol led to increased P01138 synthesis in cultured central nervous system ( CNS ) cells and rat brain tissue . DB01407 -induced P01138 expression was reduced to the control levels by coadministration of beta-adrenoceptor antagonist propranolol . Furthermore , clenbuterol protected rat hippocampal neurons subjected to excitotoxic damage . The neuroprotective effect of clenbuterol in vitro depended on increased P01138 synthesis , since the neuroprotection was abolished by P01138 antisense oligonucleotide as well as by antibodies directed against P01138 itself . In vivo , clenbuterol protected rat hippocampus in a model of transient forebrain ischemia and reduced the infarct volume in a rat model of permanent middle cerebral artery occlusion ( MCAo ) . The neuroprotective effect of clenbuterol in vivo was accompanied by enhanced P01138 synthesis in brain tissue . These findings support our hypothesis that orally active P01138 inducers may have a potential as therapeutic agents for the treatment of neurodegenerative disorders and stroke . P30559 ligands induce changes in cytoskeleton in neuroblastoma cells . Aim of the present study was to evaluate effects of ligands of oxytocin receptors on gene expression of neurofilament proteins ( nestin and microtubule-associated protein 2 ( P11137 ) ) associated with neuronal differentiation and growth factors ( brain-derived neurotrophic factor ( P23560 ) and nerve growth factor ( P01138 ) ) related to neuronal growth . Fluorescent staining of F-actin was used to observe morphology of cells . Co-treatment with oxytocin and oxytocin receptor antagonist -- atosiban -- resulted in significant increase of P11137 gene expression in SK-N-SH cells . There was no effect of oxytocin on gene expression of growth factors P23560 and P01138 . Surprisingly , oxytocin with atosiban significantly increased mRNA levels for both P23560 and P01138 . Gene expression of vasopressin receptor ( P37288 ) significantly decreased in response to vasopressin . Atosiban decreased mRNA levels for oxytocin receptor ( P30559 ) and P37288 . DB00107 significantly decreased P30559 and nestin mRNA levels and increased mRNA levels for P23560 and P01138 in U-87 MG cells . The densest recruitment of F-actin filaments was observed in apical parts of filopodia in SK-N-SH cells incubated in oxytocin presence . Present data demonstrate complex role of ligands of oxytocin receptors in regulation of gene expression of intermediate filaments and thus , oxytocin might be considered as a growth factor in neuronal type of cells . A high-sensitivity electrochemical immunosensor based on mobile crystalline material-41-polyvinyl alcohol nanocomposite and colloidal gold nanoparticles . A novel competitive immunosensor was developed as a model system using anti-human serum albumin ( HSA ) -conjugated gold nanoparticles ( AuNPs ) as an electrochemical label and mobile crystalline material-41 ( P22033 -41 ) -polyvinyl alcohol ( P32926 ) mesoporous nanocomposite as an immobilization platform . However , no attempt has yet been made to use the P22033 -41 as the supporting electrolyte for the electrosynthesis of nonconducting polymer nanocomposite . This hybrid membrane was evaluated extensively by using field emission scanning electron microscopy ( FESEM ) , cyclic voltammetry ( CV ) , and differential pulse voltammetry ( DPV ) to determine its physicochemical and electrochemical properties in immunosensor application . FESEM revealed an appropriate and stable attachment between HSA and P22033 -41 and also a dense layer deposition of P22033 -41-HSA- P32926 film onto the electrode surfaces . DPV was developed for quantitative determination of antigen in biological samples . A decrease in DPV responses was observed with increasing concentrations of HSA in standard and real samples . In optimal conditions , this immunosensor based on P22033 -41- P32926 nanocomposite film could detect HSA in a high linear range ( 0.5-200 μg ml⁻¹ ) with a low detection limit of 1 ng ml⁻¹ . The proposed method showed acceptable reproducibility , stability , and reliability and could also be applied to detect the other antigens . 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 . Impaired adipogenesis and lipolysis in the mouse upon selective ablation of the retinoid X receptor alpha mediated by a tamoxifen-inducible chimeric Cre recombinase ( Cre- P27361 ) in adipocytes . P19793 ( RXRalpha ) is involved in multiple signaling pathways , as a heterodimeric partner of several nuclear receptors . To investigate its function in energy homeostasis , we have selectively ablated the RXRalpha gene in adipocytes of 4-week-old transgenic mice by using the tamoxifen-inducible Cre- P27361 recombination system . Mice lacking RXRalpha in adipocytes were resistant to dietary and chemically induced obesity and impaired in fasting-induced lipolysis . Our results also indicate that RXRalpha is involved in adipocyte differentiation . Thus , our data demonstrate the feasibility of adipocyte-selective temporally controlled gene engineering and reveal a central role of RXRalpha in adipogenesis , probably as a heterodimeric partner for peroxisome proliferator-activated receptor gamma . Molecular evolution of the oxytocin-oxytocin receptor system in eutherians . DB00107 ( P01178 ) is a nine-amino-acid peptide hormone that is mainly released at the times of uterine contractions during parturition and milk ejection during lactation , whereas a similar peptide hormone , arginine vasopressin , primarily exerts direct antidiuretic action on the kidney and causes vasoconstriction of the peripheral vessels . The genes coding for these peptides are tandemly located on the same chromosome . A tandem duplication occurring in the common ancestor of jawed vertebrates has been proposed as responsible . In contrast to the two peptide hormones , only one oxytocin receptor ( P30559 ) but three arginine vasopressin receptors ( P37288 , P47901 , and P30518 ) are known ; these receptors probably arose from two rounds of genome duplication in the common ancestor of vertebrates . In this study , we addressed the molecular evolution of the P01178 - P30559 system in eutherians . Our analyses suggest that an amino acid change from isoleucine to lysine on the eighth site ( I8L ) of the peptide , which corresponded to a change from mesotocin to P01178 , had occurred during the common ancestral lineage of eutherians . At around the same time that the emergence of P01178 occurred , functional constraints on the P01178 receptor ( pre- P30559 ) might have relaxed , and a series of nonsynonymous substitutions might have accumulated . Only a few of these nonsynonymous substitutions might have contributed to reestablishing the molecular relationship between the P01178 ligand and its receptor , after which functional constraints on the P30559 were reinstated . Since the P01178 - P30559 system plays an important role in eutherians , the evolution of the P01178 - P30559 system was probably an essential component of the genesis of the eutherian signature . Inhibition of cervical lymph node metastasis by marimastat ( DB00786 ) in an orthotopic oral squamous cell carcinoma implantation model . Activation of matrix metalloproteinase-2 ( P08253 ) is a common event in head and neck squamous cell carcinoma . An P48449 -19 cell line , derived from human oral squamous cell carcinoma and known to metastasize to cervical lymph nodes , was implanted into the lingual margin of mice . The effect of marimastat ( DB00786 ) , a broad MMP inhibitor , on the suppression of regional cervical lymph node metastasis was evaluated with an orthotopic implantation nude mice model . Marimastat was given immediately after P48449 -19 implantation and continuously administered by an osmotic pump . The mice were divided into three groups by marimastat dose ; Group A ; 0 mg/kg/day , Group B ; 30 mg/kg/day , and Group C ; 150 mg/kg/day . Twenty-one days after implantation , primary oral tumors and cervical lymph nodes were resected . Cervical lymph node status was microscopically examined . Activation of P08253 in primary oral tumor was examined by gelatin zymography . Both cervical lymph node metastasis and activation of P08253 were significantly suppressed in Group C ( P < 0.05 ) . Moreover , the Group C mice had a significantly better survival than group A ( P = 0.0026 ) . There was a significant difference between Group A and Group C in terms of proliferation of tumor cells by proliferating cell nuclear antigen immunostaining ( P = 0.0120 ) . These results suggest a positive role for marimastat in the inhibition of P08253 activation and prevention of cervical lymph node metastasis in oral squamous cell carcinoma ( OSCC ) . Improvement of survival in patients with OSCC could be expected using adjuvant therapy with marimastat . P04035 inhibition reduces the proinflammatory activation of human vascular smooth muscle cells by the terminal complement factor C5b-9 . The terminal complement complex C5b-9 is known to participate in inflammatory processes including atherosclerosis . Inflammation appears to be a direct consequence of C5b-9-mediated cell stimulation . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors may exert anti-inflammatory effects on vascular cells independent of lowering plasma cholesterol . Thus , we studied activation of vascular smooth muscle cells ( VSMCs ) by C5b-9 focusing on whether inhibition of the P04035 can reduce the proinflammatory effects of C5b-9.C5b-9 in sublytic concentrations increased the proliferation of human VSMCs and induced a time-dependent activation of the mitogen-activated protein ( Q96HU1 ) kinase extracellular signal-regulated kinase ( P29323 ) . Proliferation and P27361 /2 activation could be inhibited by the specific P29323 inhibitor PD98059 . HMG- DB01992 inhibition with cerivastatin-reduced VSMC proliferation and C5b-9-induced P27361 /2 activation . DB00439 also reduced the C5b-9-induced synthesis of the proinflammatory interleukin-6 ( P05231 ) . Furthermore , C5b-9 induced activation of the transcription factors activator protein- 1 ( AP-1 ) and nuclear factor-kappaB ( NF-kappaB ) , which could be inhibited by pretreatment of VSMCs with cerivastatin . L-mevalonate and geranylgeranylpyrophosphate reversed the inhibitory effects of cerivastatin . The present study in VSMCs shows that cerivastatin inhibits P05231 synthesis and cell proliferation induced by the terminal complement complex C5b-9 . This may be an important mechanism contributing to the beneficial effects of P04035 inhibitors beyond lowering of plasma cholesterol . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . Selective measurement of white matter and gray matter diffusion trace values in normal human brain . The trace of the diffusion tensor ( or simply the trace ) is diagnostically valuable for detecting acute ischemic lesions . A number of studies indicate that the trace of human gray matter ( GM ) and white matter ( WM ) are quite similar . This is somewhat surprising considering the different cellular environments of GM and WM . It is possible that partial volume averaging ( P32926 ) effects between GM and WM , inherent in many of the ultrafast imaging sequences used for diffusion measurements , are responsible for this observation . In order to minimize P32926 effects , the trace values of GM and WM have been selectively measured by implementing double inversion recovery ( P30518 ) echo planar imaging ( P10646 ) pulse sequences . Results on six normal volunteers indicate that the trace values of WM and GM are not statistically different . An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the Q7RTX0 taste receptor . Sweet taste is mediated by a dimeric receptor composed of two distinct subunits , Q8TE23 and Q7RTX0 , whereas the Q7RTX1 / Q7RTX0 receptor is involved in umami taste perception . The Q7RTX1 , Q8TE23 , and Q7RTX0 subunits are members of the small family of class C G protein-coupled receptors ( GPCRs ) . The members of this family are characterized by a large N-terminal domain ( NTD ) , which is structurally similar to bacterial periplasmic-binding proteins and contains the primary ligand-binding site . In a recent study , we described a strategy to produce a functional dimeric human Q7RTX0 -NTD . Although the protein was expressed as inclusion bodies ( IBs ) using the Escherichia coli system , the conditions for the refolding of functional hT1R3-NTD were determined using a fractional factorial screen coupled to a binding assay . Here , we report that this refolding strategy can be used to produce Q7RTX1 - and Q8TE23 -NTDs in large quantities . We also discuss that our findings could be more generally applicable to other class C GPCR-NTDs , including the γ-aminobutyric acid type B receptor ( GABABR ) , the extracellular calcium-sensing receptor ( P41180 ) and the large family of pheromone ( P30518 ) orphan receptors . 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 . Q9GZV9 and its receptors . Q9GZV9 ( Q9GZV9 ) is a circulating factor that plays critical roles in phosphate and vitamin D metabolism , as evidenced by the fact that Q9GZV9 missense mutations cause autosomal dominant hypophosphatemic rickets ( P30518 ) . Autosomal dominant hypophosphatemic rickets is characterized by hypophosphatemia with inappropriately normal 1,25-dihydroxyvitamin D concentrations , as well as bone pain , fracture and rickets . This phenotype parallels that of patients with tumor induced osteomalacia ( TIO ) , X-linked hypophosphatemic rickets ( XLH ) , and fibrous dysplasia ( FD ) , in whom elevated serum Q9GZV9 levels are often observed . The fibroblast growth factor receptors ( P11362 -4 ) play key roles in skeletal development , as well as in normal metabolic processes . Several FGFR isoforms that potentially mediate the activity of Q9GZV9 have been implicated . In the short term , these findings will lead to further understanding of Q9GZV9 function , and potentially in the long term , to targeted therapies in disorders of hypo- and hyperphosphatemia that involve Q9GZV9 . 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 . Angiotensin II blockade : how its molecular targets may signal to mitochondria and slow aging . Coincidences with calorie restriction and P42345 inhibition . Caloric restriction ( CR ) , renin angiotensin system blockade ( DB01367 -bl ) , and rapamycin-mediated mechanistic target of rapamycin ( P42345 ) inhibition increase survival and retard aging across species . Previously , we have summarized CR and DB01367 -bl 's converging effects , and the mitochondrial function changes associated with their physiological benefits . P42345 inhibition and enhanced sirtuin and KLOTHO signaling contribute to the benefits of CR in aging . mTORC1/mTORC2 complexes contribute to cell growth and metabolic regulation . Prolonged mTORC1 activation may lead to age-related disease progression ; thus , rapamycin-mediated P42345 inhibition and CR may extend lifespan and retard aging through mTORC1 interference . Sirtuins by deacetylating histone and transcription-related proteins modulate signaling and survival pathways and mitochondrial functioning . CR regulates several mammalian sirtuins favoring their role in aging regulation . KLOTHO/fibroblast growth factor 23 ( Q9GZV9 ) contribute to control Ca(2+) , phosphate , and vitamin D metabolism , and their dysregulation may participate in age-related disease . Here we review how P42345 inhibition extends lifespan , how KLOTHO functions as an aging suppressor , how sirtuins mediate longevity , how vitamin D loss may contribute to age-related disease , and how they relate to mitochondrial function . Also , we discuss how DB01367 -bl downregulates P42345 and upregulates KLOTHO , sirtuin , and vitamin D receptor expression , suggesting that at least some of DB01367 -bl benefits in aging are mediated through the modulation of P42345 , KLOTHO , and sirtuin expression and vitamin D signaling , paralleling CR actions in age retardation . Concluding , the available evidence endorses the idea that DB01367 -bl is among the interventions that may turn out to provide relief to the spreading issue of age-associated chronic disease .

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Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . An altered fibronectin matrix induces anoikis of human squamous cell carcinoma cells by suppressing integrin alpha v levels and phosphorylation of Q05397 and P29323 . P02751 regulates many cellular processes , including migration , proliferation , differentiation , and survival . Previously , we showed that squamous cell carcinoma ( SCC ) cell aggregates escape suspension-induced , p53-mediated anoikis by engaging in fibronectin-mediated survival signals through focal adhesion kinase ( Q05397 ) . Here we report that an altered matrix , consisting of a mutated , nonfunctional high-affinity heparin-binding domain and the V region of fibronectin ( V+H- ) , induced anoikis in human SCC cells ; this response was blocked by inhibitors of caspase-8 and caspase-3 . Anoikis was mediated by downregulation of integrin alpha v in a panel of SCC cells and was shown to be proteasome-dependent . Overexpression of integrin alpha v or Q05397 inhibited the increase in caspase-3 activation and apoptosis , whereas suppression of alpha v or Q05397 triggered a further significant increase in apoptosis , indicating that the apoptosis was mediated by suppression of integrin alpha v levels and dephosphorylation of Q05397 . Treatment with V+H- decreased the phosphorylation of extracellular signal-regulated kinase ( P29323 ) 1 and 2 , and direct activation of P29323 by constitutively active Q02750 , an P29323 kinase , increased P27361 and P28482 phosphorylation and inhibited the increase in apoptosis induced by V+H- . P29323 acted downstream from alpha v and Q05397 signals , since alpha v and Q05397 overexpression inhibited both the decrease in P29323 phosphorylation and the increase in anoikis triggered by V+H- . These findings provide evidence that mutations in the high-affinity heparin-binding domain in association with the V region of fibronectin , or altered fibronectin matrices , induce anoikis in human SCC cells by modulating integrin alpha v-mediated phosphorylation of Q05397 and P29323 . [ Regulation of P04271 expression during long term potentiation ] . In this study , contributions of intracellular regulatory cascades in the induction of P04271 expression in rat hippocampal P00915 area during long term posttetanic potentiation ( LTP ) were estimated . The activation of transcription factor p53 ( positive regulator of P04271 transcription ) by nutlin-3 increased the basal content of P04271 mRNA up to 151 % of the control level , which was significantly lower than its content in tetanized slices ( 280 % ) . Therefore , p53 seems to be not unique transcription factor upregulating P04271 expression during LTP . The inhibitor of Ca2+/calmodulin-dependent kinases ( CaMKs ) KN-93 fully blocked the increase of P04271 mRNA after tetanization , while KN-92 ( inactive analogue of KN-93 ) was ineffective . The inhibitor of CaMKII and receptor tyrosine kinases DB02152 essentially suppressed P04271 expression during LTP , the inhibition of MAPK p38 or P51812 moderately decreased , and the inhibition of Q02750 did not influence P04271 mRNA content . Thus , CaMKs play a key role in the induction of P04271 expression during LTP . Tumor angiogenesis mediated by myeloid cells is negatively regulated by P13688 . Bv8 ( prokineticin 2 ) expressed by Gr1(+)CD11b(+) myeloid cells is critical for P15692 -independent tumor angiogenesis . Although granulocyte colony-stimulating factor ( DB00099 ) has been shown to be a key inducer of Bv8 expression , the basis for Bv8 production in driving tumor angiogenesis is undefined . Because the cell adhesion molecule P13688 , which is highly expressed on Gr1(+)CD11b(+) myeloid cells , is known to regulate Q99062 ( G-CSFR ) signaling , we hypothesized that P13688 would regulate Bv8 production in these cells . In support of this hypothesis , we found that Bv8 expression was elevated in Gr1(+)CD11b(+) cells from Ceacam1-deficient mice implanted with B16 melanoma , increasing the infiltration of Gr1(+)CD11b(+) myeloid cells in melanoma tumors and enhancing their growth and angiogenesis . Furthermore , treatment with anti-Gr1 or anti-Bv8 or anti-G- P04141 monoclonal antibody reduced myeloid cell infiltration , tumor growth , and angiogenesis to levels observed in tumor-bearing wild-type ( WT ) mice . Reconstitution of P13688 -deficient mice with WT bone marrow cells restored tumor infiltration of Gr1(+)CD11b(+) cells along with tumor growth and angiogenesis to WT levels . Treatment of tumor-bearing WT mice with anti- P13688 antibody limited tumor outgrowth and angiogenesis , albeit to a lesser extent . Tumor growth in Ceacam1-deficient mice was not affected significantly in Rag(-/-) background , indicating that P13688 expression in T and B lymphocytes had a negligible role in this pathway . Together , our findings show that P13688 negatively regulates Gr1(+)CD11b(+) myeloid cell-dependent tumor angiogenesis by inhibiting the G- P04141 -Bv8 signaling pathway . 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 . 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 . 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 . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . 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 . Linkage assignment of eleven genes to the porcine genome . We report comparative linkage mapping of eleven genes in the swine genome by RFLP analysis . These genes include : Acid phosphatase type 5 ( P13686 ) , Cholecystokinin Type B Receptor ( P32239 ) , Antibiotic Peptide ( P49913 ) , P01308 -like Growth Factor 1 Receptor ( P08069 ) , Integrin Alpha M ( P11215 ) , Integrin Beta 2 ( ITGbeta2 ) , Opioid Receptor Mu-1 ( P35372 ) , Pro-hormone Converter ( PC1/3 ) , DB00162 Binding Q12988 ( P10745 ) , Ribosomal DNA ( RNR1 ) , and Zona Pellucida Glycoprotein 1 ( P60852 ) . The P32239 and ITGbeta2 loci define the ends of the linkage groups on Chromosomes ( Chro ) ( SSC ) 9p and 13qter , respectively . Polymorphisms of dopamine receptor/transporter genes and risk of non-small cell lung cancer . BACKGROUND : The dopaminergic pathway may be of interest in assessing risk of non-small cell lung cancer ( NSCLC ) . Dopamine receptors are expressed in alveolar epithelial cells and human lung tumours , and dopamine inhibits both cell proliferation in vitro and growth of lung tumour xenografts in nude mice . Moreover , dopamine selectively inhibits the vascular permeability and angiogenic activity of vascular endothelial growth factor ( P15692 / P15692 ) . The bioavailability of dopamine is regulated by dopamine receptors D2 ( P14416 ) , D4 ( P21917 ) and dopamine transporter 1 ( Q01959 / Q01959 ) genes . METHODS : We have analysed 10 single nucleotide polymorphisms in P14416 , P21917 and Q01959 / Q01959 genes in relation to lung cancer risk in a case-control study of smoking subjects . The study subjects were 413 healthy individuals from general population and 335 NSCLC cases . Both cases and controls were Caucasians of Norwegian origin . RESULTS : We demonstrate that P14416 polymorphisms -141Cdel , 3208G > T , TaqIB ; P21917 -521C > T and Q01959 / Q01959 -1476T > G are associated with a two- to five-fold increased NSCLC risk . The variant alleles of P14416 1412A > G and 960C > G had protective effects . CONCLUSION : The dopamine receptor/transport gene polymorphisms are associated with the risk of NSCLC among smokers . The data show that the polymorphisms resulting in lower dopamine bioavailability were associated with increased risk of NSCLC . 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 . 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 . 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 . MicroRNA profiling of novel African American and Caucasian Prostate Cancer cell lines reveals a reciprocal regulatory relationship of miR-152 and DNA methyltranferase 1 . miRNA expression in African American compared to Caucasian PCa patients has not been widely explored . Herein , we probed the miRNA expression profile of novel AA and CA derived prostate cancer cell lines . We found a unique miRNA signature associated with AA cell lines , independent of tumor status . Evaluation of the most differentially expressed miRNAs showed that miR-132 , miR-367b , miR-410 , and miR-152 were decreased in more aggressive cells , and this was reversed after treatment of the cells with DB01262 . Sequencing of the miR-152 promoter confirmed that it was highly methylated . Ectopic expression of miR-152 resulted in decreased growth , migration , and invasion . Informatics analysis of a large patient cohort showed that decreased miR-152 expression correlated with increased metastasis and a decrease in biochemical recurrence free survival . Analysis of 39 prostate cancer tissues with matched controls ( 20 AA and 19 CA ) , showed that 50 % of AA patients had statistically significant lower miR-152 expression compared to only 35 % of CA patients . Ectopic expression of miR-152 in LNCaP , PC-3 , and MDA-PCa-2b cells down-regulated P26358 ( P26358 ) through direct binding in the P26358 3'UTR . There appeared to be a reciprocal regulatory relationship of miR-152/ P26358 expression , as cells treated with siRNA P26358 caused miR-152 to be re-expressed in all cell lines . In summary , these results demonstrate that epigenetic regulation of miR-152/ P26358 may play an important role in multiple events that contribute to the aggressiveness of PCa tumors , with an emphasis on AA PCa patients . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Transient multiple acyl- DB01992 dehydrogenation deficiency in a newborn female caused by maternal riboflavin deficiency . A newborn female presented on the first day of life with clinical and biochemical findings consistent with multiple acyl- DB01992 dehydrogenase deficiency ( Q8WXG6 ) . DB00140 supplementation corrected the biochemical abnormalities 24 h after commencing the vitamin . In vitro acylcarnitine profiling in intact fibroblasts both in normal and riboflavin depleted media showed normal oxidation of fatty acids excluding defects in electron transfer flavoprotein ( ETF ) , or ETF ubiquinone oxidoreductase ( ETF:QO ) , or a genetic abnormality in flavin metabolism . In addition , sequencing of the genes encoding ETF and ETF:QO in the proband did not reveal any pathogenic mutations . Determination of the maternal riboflavin status after delivery showed that the mother was riboflavin deficient . Repeat testing done two years after the infant 's birth and while on a normal diet showed that the mother was persistently riboflavin deficient and showed a typical Q8WXG6 profile on plasma acylcarnitine testing . A possible genetic defect in riboflavin transport of metabolism in the mother is postulated to be the cause of the transient Q8WXG6 seen in the infant . Sequencing of the Q6ZSM3 , Q969G6 and Q8NFF5 genes encoding key enzymes in riboflavin transport of metabolism in the mother did not identify any pathogenic mutations . The underlying molecular basis of the mother 's defect in riboflavin metabolism remains to be established . DB00183 infusions in patients with panic disorder . I . Symptoms and cardiovascular responses . Cholecystokinin ( CCK ) may mediate human anxiety and animal data suggest that cholecystokinin antagonists could provide an important advance in the treatment of anxiety disorders . The study of CCK receptor systems in psychiatric patients has , however , been severely limited by the lack of available probes . We utilized intravenous infusions of pentagastrin , a selective P32239 agonist , and studied behavioral and cardiovascular responses in 10 patients with panic disorder and 10 normal controls . DB00183 produced substantial symptomatology , including anxiety , and increases in heart rate and blood pressure , in both patients and controls . Patients were more sensitive to the panicogenic effects of the pentagastrin . Panic attacks occurred in 70 % of patients and 0 % of controls . Patients ' symptom responses were very similar to their " typical " panic attacks and to symptoms produced by Q13308 . DB00183 provides a readily available alternative to Q13308 for studying the CCK receptor system and exploring its involvement in human anxiety . DB08954 induced antinociception and decreased the expression of Q13224 subunits in the dorsal horn after chronic dorsal root ganglia compression in rats . BACKGROUND : Spinal N-methyl D-aspartate receptors play an important role in the pathogenesis of neuropathic pain , and administration of N-methyl D-aspartate receptor antagonists can attenuate this hyperpathia . DB08954 is an antagonist selective for N-methyl D-aspartate receptor 2B ( Q13224 ) subunits . Several researches have reported effective analgesia of ifenprodil in animal models of neuropathic pain . We extended this work to include chronic compression of the dorsal root ganglia ( CCD ) . METHODS : The paw withdrawal mechanical threshold and paw withdrawal thermal latency tests were used to assess mechanical allodynia and thermal hyperalgesia after a CCD operation and intrathecal injection of ifenprodil . We used immunohistochemistry and immunoblotting to investigate the effect of ifenprodil on Q13224 subunits expression in CCD rats . RESULTS : The data revealed increased expression of Q13224 subunits in the superficial dorsal horn in CCD rats . We found that , in addition to a marked suppression of thermal hyperalgesia and mechanical allodynia , intrathecal injection ifenprodil treatment causes a decreased expression of Q13224 in the spinal cord . CONCLUSIONS : These data suggest that ifenprodil induced antinociception in CCD rats and provided further evidence for the important role of Q13224 subunits in the development of neuropathic pain . 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 . Epigenetic control of skin differentiation genes by phytocannabinoids . BACKGROUND AND PURPOSE : Endocannabinoid signalling has been shown to have a role in the control of epidermal physiology , whereby anandamide is able to regulate the expression of skin differentiation genes through DNA methylation . Here , we investigated the possible epigenetic regulation of these genes by several phytocannabinoids , plant-derived cannabinoids that have the potential to be novel therapeutics for various human diseases . EXPERIMENTAL APPROACH : The effects of cannabidiol , cannabigerol and cannabidivarin on the expression of skin differentiation genes keratins 1 and 10 , involucrin and transglutaminase 5 , as well as on DNA methylation of keratin 10 gene , were investigated in human keratinocytes ( HaCaT cells ) . The effects of these phytocannabinoids on global DNA methylation and the activity and expression of four major DNA methyltransferases ( P26358 , 3a , 3b and 3L ) were also examined . KEY RESULTS : DB09061 and cannabigerol significantly reduced the expression of all the genes tested in differentiated HaCaT cells , by increasing DNA methylation of keratin 10 gene , but cannabidivarin was ineffective . Remarkably , cannabidiol reduced keratin 10 mRNA through a type-1 cannabinoid ( P21554 ) receptor-dependent mechanism , whereas cannabigerol did not affect either P21554 or CB2 receptors of HaCaT cells . In addition , cannabidiol , but not cannabigerol , increased global DNA methylation levels by selectively enhancing P26358 expression , without affecting P26358 3a , 3b or 3L . CONCLUSIONS AND IMPLICATIONS : These findings show that the phytocannabinoids cannabidiol and cannabigerol are transcriptional repressors that can control cell proliferation and differentiation . This indicates that they ( especially cannabidiol ) have the potential to be lead compounds for the development of novel therapeutics for skin diseases .

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interacts_with DB00472?

multiple_choice

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 . 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 . P00734 kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation . We have shown that prothrombin kringle-2 ( pKr-2 ) , a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro . However , little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic ( DA ) neurons in vivo . To address this question , pKr-2 was injected into the rat substantia nigra ( SN ) . Tyrosine hydroxylase ( TH ) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2 . In parallel , pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry . Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-2 ( P35354 ) and several proinflammatory cytokines . The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride , and the P35354 inhibitor DuP-697 . P27361 /2 , c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection , and localized within microglia . Inhibition of these kinases led to attenuation of mRNA expression of P35228 , P35354 and several proinflammatory cytokines , and rescue of DA neurons in the SN . Intriguingly , following treatment with pKr-2 in vitro , neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia , but not microglia-free neuron-enriched mesencephalic cultures , indicating that microglia are required for pKr-2 neurotoxicity . Our results strongly suggest that microglia activated by endogenous compound(s) , such as pKr-2 , are implicated in the DA neuronal cell death in the SN . The molecular mechanism of the inhibition by licofelone of the biosynthesis of P09917 products . BACKGROUND AND PURPOSE : DB04725 is a dual inhibitor of the cyclooxygenase and P09917 ( P09917 ) pathway , and has been developed for the treatment of inflammatory diseases . Here , we investigated the molecular mechanisms underlying the inhibition by licofelone of the formation of P09917 products . EXPERIMENTAL APPROACH : The efficacy of licofelone to inhibit the formation of P09917 products was analysed in human isolated polymorphonuclear leukocytes ( PMNL ) or transfected HeLa cells , as well as in cell-free assays using respective cell homogenates or purified recombinant P09917 . Moreover , the effects of licofelone on the subcellular redistribution of P09917 were studied . KEY RESULTS : DB04725 potently blocked synthesis of P09917 products in Ca(2+)-ionophore-activated PMNL ( IC(50)=1.7 microM ) but was a weak inhibitor of P09917 activity in cell-free assays ( IC(50) >> 10 microM ) . The structures of licofelone and MK-886 , an inhibitor of the P09917 -activating protein ( P20292 ) , were superimposable . The potencies of both licofelone and MK-886 in ionophore-activated PMNL were impaired upon increasing the concentration of arachidonic acid , or under conditions where P09917 product formation was evoked by genotoxic , oxidative or hyperosmotic stress . Furthermore , licofelone prevented nuclear redistribution of P09917 in ionophore-activated PMNL , as had been observed for P20292 inhibitors . Finally , licofelone as well as MK-886 caused only moderate inhibition of the synthesis of P09917 products in HeLa cells , unless P20292 was co-transfected . CONCLUSIONS AND IMPLICATIONS : Our data suggest that the potent inhibition of the biosynthesis of P09917 products by licofelone requires an intact cellular environment and appears to be due to interference with P20292 . 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 . Prostaglandin synthase 2/cyclooxygenase 2 ( P35354 / P35354 ) 8473T > C polymorphism associated with prognosis for patients with colorectal cancer treated with capecitabine and oxaliplatin . PURPOSE : The present study analyzed the polymorphisms of apoptosis-related genes and their impact on the response to chemotherapy and survival of patients with colorectal cancer . PATIENTS AND METHODS : A total of 76 patients with recurrent or metastatic colorectal cancer treated with capecitabine and oxaliplatin ( XELOX ) combination chemotherapy were enrolled in the present study . The single nucleotide polymorphisms of 15 apoptosis-related genes ( P04637 , Q07817 , O14763 , P31749 , P35354 / P35354 , P55957 , Q13546 , FAS , P48023 , caspase 3 , and caspase 6-10 ) were determined using a PCR-RFLP assay . RESULTS : No significant association between the polymorphisms and the response was found for any of the genes analyzed . However , the T/T genotype of P35354 8473T > C ( rs5275 ) was significantly correlated with a better progression-free survival ( PFS ) and overall survival ( OS ) when compared to the combined T/C and C/C genotype ( Hazard ratio [ HR ] = 0.47 ; P value = 0.046 and HR = 0.16 ; P = 0.013 , respectively ) in a multivariate analysis adjusted for age , sex , performance status , disease status and curative resection . No association was noted between the other polymorphisms and survival . CONCLUSION : The P35354 8473T > C polymorphism was found to be correlated with PFS and OS in patients with advanced colorectal cancer treated with XELOX chemotherapy . Serotonin and fluoxetine receptors are expressed in enamel organs and LS8 cells and modulate gene expression in LS8 cells . The selective serotonin re-uptake inhibitor ( SSRI ) fluoxetine is widely used in the treatment of depression in children and fertile women , but its effect on developing tissues has been sparsely investigated . The aim of this study was to investigate if enamel organs and ameloblast-derived cells express serotonin receptors that are affected by peripherally circulating serotonin or fluoxetine . Using RT-PCR and western blot analysis we found that enamel organs from 3-d-old mice and ameloblast-like cells ( LS8 cells ) express functional serotonin receptors , the rate-limiting enzyme in serotonin synthesis ( Thp1 ) , as well as the serotonin transporter ( P31645 ) , indicating that enamel organs and ameloblasts are able to respond to serotonin and regulate serotonin availability . DB00472 and serotonin enhanced the alkaline phosphatase activity in the cell culture medium from cultured LS8 cells , whereas the expression of enamelin ( Enam ) , amelogenin ( Amel ) , and matrix metalloproteinase-20 ( O60882 ) were all significantly down-regulated . The secretion of vascular endothelial growth factor ( P15692 ) , monocyte chemotactic protein 1 ( P13500 ) , and interferon-inducible protein 10 ( P02778 ) was also reduced compared with controls . In conclusion , enamel organs and ameloblast-like cells express functional serotonin receptors . Reduced transcription of enamel proteins and secretion of vascular factors may indicate possible adverse effects of fluoxetine on amelogenesis . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . 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 . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . P01308 signaling inhibits the P28335 receptor in choroid plexus via Q96HU1 kinase . BACKGROUND : G protein-coupled receptors ( GPCRs ) interact with heterotrimeric GTP-binding proteins ( G proteins ) to modulate acute changes in intracellular messenger levels and ion channel activity . In contrast , long-term changes in cellular growth , proliferation and differentiation are often mediated by tyrosine kinase receptors and certain GPCRs by activation of mitogen-activated protein ( Q96HU1 ) kinases . Complex interactions occur between these signaling pathways , but the specific mechanisms of such regulatory events are not well-understood . In particular it is not clear whether GPCRs are modulated by tyrosine kinase receptor- Q96HU1 kinase pathways . RESULTS : Here we describe tyrosine kinase receptor regulation of a GPCR via Q96HU1 kinase . P01308 reduced the activity of the P28335 receptor in choroid plexus cells which was blocked by the Q96HU1 kinase kinase ( MEK ) inhibitor , PD 098059 . We demonstrate that the inhibitory effect of insulin and insulin-like growth factor type 1 ( DB01277 ) on the P28335 receptor is dependent on tyrosine kinase , DB01367 and Q96HU1 kinase . The effect may be receptor-specific : insulin had no effect on another GPCR that shares the same G protein signaling pathway as the P28335 receptor . This effect is also direct : activated Q96HU1 kinase mimicked the effect of insulin , and removing a putative Q96HU1 kinase site from the P28335 receptor abolished the effect of insulin . CONCLUSION : These results show that insulin signaling can inhibit P28335 receptor activity and suggest that Q96HU1 kinase may play a direct role in regulating the function of a specific GPCR . 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 . P04271 /RAGE-dependent activation of microglia via NF-kappaB and AP-1 Co-regulation of P35354 expression by P04271 , IL-1beta and P01375 . Extracellular P04271 is known to affect astrocytic , neuronal and microglial activities , with different effects depending on its concentration . Whereas at relatively low concentrations P04271 exerts trophic effects on neurons and astrocytes , at relatively high concentrations the protein causes neuronal apoptosis and activates astrocytes and microglia , thus potentially representing an endogenous factor implicated in neuroinflammation . We have reported that RAGE ligation by P04271 in BV-2 microglia results in the upregulation of expression of the pro-inflammatory cyclo-oxygenase 2 ( P35354 ) via parallel Ras-Cdc42-Rac1-dependent activation of c-Jun NH(2) terminal protein kinase ( JNK ) and Ras-Rac1-dependent stimulation of NF-kappaB transcriptional activity . We show here that : ( 1 ) P04271 also stimulates AP-1 transcriptional activity in microglia via RAGE-dependent activation of JNK ; ( 2 ) P04271 upregulates IL-1beta and P01375 expression in microglia via RAGE engagement ; and ( 3 ) P04271 /RAGE-induced upregulation of P35354 , IL-1beta and P01375 expression requires the concurrent activation of NF-kappaB and AP-1 . We also show that P04271 synergizes with IL-1beta and P01375 to upregulate on P35354 expression in microglia . Given the crucial roles of P35354 , IL-1beta and P01375 in the inflammatory response , we propose that , by engaging RAGE , P04271 might play an important role in microglia activation in the course of brain damage . Population-specific genetic associations with oesophageal squamous cell carcinoma in South Africa . Genetic variants in multiple cellular pathways have been associated with an altered risk of oesophageal cancer . In this study , eight genes previously associated with an altered risk of oesophageal squamous cell carcinoma ( OSCC ) in European or Asian populations were investigated in two South African populations . We genotyped 12 single-nucleotide polymorphisms and one insertion/deletion variant in 1463 individuals from the Black and Mixed Ancestry populations . No polymorphisms were associated with OSCC in the Black population . In the Mixed Ancestry population , P05091 +82 G > A ( rs886205 ) was significantly associated with a reduced risk of OSCC ( odds ratio = 0.70 , 95 % confidence interval = 0.55-0.89 ; P = 0.0038 ) . Several other polymorphisms showed a suggestive association ( P < 0.05 ) , including P00325 Arg48His ( rs1229984 ) , P35354 -1195G > A ( rs689466 ) , Q14790 Asp302His ( rs1045485 ) and P16455 Leu84Phe ( rs12917 ) . Haplotype analysis indicated that the FAS polymorphisms -670 A > G ( rs1800682 ) and -1377 G > A ( rs2234767 ) were both associated with OSCC in the Mixed Ancestry population ( P = 0.006 and P = 0.004 , respectively ) , as well as the Q14790 ( -652 6Ndel:302His ) haplotype ( P = 0.0013 ) . This study indicates several instances of population-specific differences in the genetic etiology of OSCC between these two South African populations and between them and other high-risk populations , which may reflect differences in their ancestry and environmental exposures . 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 . Characterization of the activity of the PI3K/ P42345 inhibitor DB05241 ( SAR245409 ) in tumor models with diverse genetic alterations affecting the PI3K pathway . Activation of the PI3K ( phosphoinositide 3-kinase ) pathway is a frequent occurrence in human tumors and is thought to promote growth , survival , and resistance to diverse therapies . Here , we report pharmacologic characterization of the pyridopyrimidinone derivative DB05241 ( SAR245409 ) , a potent and highly selective pan inhibitor of class I PI3Ks ( α , β , γ , and δ ) with activity against P42345 . Broad kinase selectivity profiling of > 130 protein kinases revealed that DB05241 is highly selective for class I PI3Ks and P42345 over other kinases . In cellular assays , DB05241 inhibits the formation of PIP(3) in the membrane , and inhibits phosphorylation of AKT , p70S6K , and S6 phosphorylation in multiple tumor cell lines with different genetic alterations affecting the PI3K pathway . In a panel of tumor cell lines , DB05241 inhibits proliferation with a wide range of potencies , with evidence of an impact of genotype on sensitivity . In mouse xenograft models , oral administration of DB05241 results in dose-dependent inhibition of phosphorylation of AKT , p70S6K , and S6 with a duration of action of approximately 24 hours . Repeat dose administration of DB05241 results in significant tumor growth inhibition in multiple human xenograft models in nude mice that is associated with antiproliferative , antiangiogenic , and proapoptotic effects . Perfusion-independent effect of bradykinin and fosinoprilate on glucose transport in Langendorff rat hearts . P12821 ( P12821 ) inhibitor-stimulated glucose metabolism and perfusion in muscle tissue seem to be , at least in part , mediated by kinins . However , the relative contribution of direct metabolic or secondary hemodynamically induced effects is unclear . It was the aim of this study to characterize the effects of P12821 inhibition and bradykinin on glucose transport while changes in cardiocoronary function that might influence glucose transport were minimized . Hearts from Wistar rats were perfused by a Langendorff preparation and a set of functional parameters were simultaneously measured . Bradykinin ( 10[-11] M ) and fosinoprilate ( 10[-7] M ) were administered at concentrations that did not affect coronary flow . P01308 was employed as reference at half-maximal concentration . The nonmetabolizable glucose analog 3-O-[14C]methyl-D-glucose and the nontransportable tracer L-[3H]glucose were coperfused for the calculation of glucose transport . Using a 2-compartment mathematical model we found that the glucose transport rate , which was doubled with insulin , was increased almost 3-fold by either bradykinin or fosinoprilate . In the presence of the P30411 antagonist DB06196 ( D- DB00125 [Hyp3,Thi5,D-Tic7,Oic8]-bradykinin ; icatibant ) , the effect of both agents was completely abolished . Both agents also induced minor changes in contractility/relaxation parameters that again were completely neutralized with icatibant . A perfusion-independent but B2-kinin receptor-dependent stimulating effect on glucose transport by either bradykinin or fosinoprilate is concluded . This effect could , in analogy to insulin be due to increased glucose transporter translocation , increased endothelium-derived nitric oxide formation , or -- despite constant coronary flow conditions -- secondary to altered cardiac function . P04141 -1 ( P09603 ) delivers a proatherogenic signal to human macrophages . P09603 / P09603 supports the proliferation and differentiation of monocytes and macrophages . In mice , P09603 also promotes proinflammatory responses in vivo by regulating mature macrophage functions , but little is known about the acute effects of this growth factor on mature human macrophages . Here , we show that in contrast to its effects on mouse bone marrow-derived macrophages , P09603 did not induce expression of urokinase plasminogen activator mRNA , repress expression of apolipoprotein E mRNA , or prime LPS-induced P01375 and P05231 secretion in human monocyte-derived macrophages ( HMDM ) from several independent donors . Instead , we show by expression profiling that P09603 modulates the HMDM transcriptome to favor a proatherogenic environment . P09603 induced expression of the proatherogenic chemokines P02778 /IFN-inducible protein 10 , P13500 , and P80098 but repressed expression of the antiatherogenic chemokine receptor P61073 . P09603 also up-regulated genes encoding enzymes of the cholesterol biosynthetic pathway ( P04035 , P53602 , Q13907 , P14324 , Q14534 , Q16850 , EBP , Q15738 , Q9UBM7 , and Q15392 ) , and expression of P45844 , encoding a cholesterol efflux transporter , was repressed . Consistent with these effects , P09603 increased levels of free cholesterol in HMDM , and the selective P07333 kinase inhibitor GW2580 ablated this response . These data demonstrate that P09603 represents a further link between inflammation and cardiovascular disease and suggest two distinct mechanisms by which P09603 , which is known to be present in atherosclerotic lesions , may contribute to plaque progression . 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 .

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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 . P07550 mediated P29323 activation is regulated by interaction with Q5TCQ9 . The beta-2 adrenergic receptor ( beta2AR ) has a carboxyl terminus motif that can interact with P78352 /discs-large/ Q07157 homology ( PDZ ) domain-containing proteins . In this paper , we identified membrane-associated guanylate kinase inverted-3 ( Q5TCQ9 ) as a novel binding partner of beta2AR . The carboxyl terminus of beta2AR binds with high affinity to the fifth PDZ domain of Q5TCQ9 , with the last four amino acids ( D-S-L-L ) of the receptor being the key determinants of the interaction . In cells , the association of full-length beta2AR with Q5TCQ9 occurs constitutively and is enhanced by agonist stimulation of the receptor . Our data also demonstrated that beta2AR-stimulated extracellular signal-regulated kinase-1/2 ( P27361 /2 ) activation was substantially retarded by Q5TCQ9 expression . These data suggest that Q5TCQ9 regulates beta2AR-mediated P29323 activation through the physical interaction between beta2AR and Q5TCQ9 . A new epigenetic challenge : systemic lupus erythematosus . In recent years , compelling evidence has been gathered that supports a role for epigenetic alterations in the pathogenesis of systemic lupus erythematosus ( SLE ) . Different blood cell populations of SLE patients are characterized by a global loss of DNA methylation . This process is associated with defects in P29323 pathway signalling and consequent P26358 1 downregulation . Hypomethylation of gene promoters has been described , which permits transcriptional activation and therefore functional changes in the cells and also hypomethylation of the ribosomal RNA gene cluster . Among the identified targets undergoing demethylation are genes involved in autoreactivity ( P20701 ) , osmotic lysis and apoptosis ( P14222 , P50281 and P80188 ) , antigen presentation ( Q99062 ) , inflammation ( MMP 14 ) , B- T-cell interaction ( P32970 and P29965 ) and cytokine pathways ( Q99062 , P05112 , P05231 and P38484 ) . DNA methylation inhibitors are also known to induce autoreactivity in vitro and cause a lupus-like disease in vivo . Further , altered patterns of histone modifications have been described in SLE . P01730 + lymphocytes undergo global histone H3 and H4 deacetylation and consequent skewed gene expression . Although multiple lines of evidence highlight the contribution of epigenetic alterations to the pathogenesis of lupus in genetically predisposed individuals , many questions remain to be answered . Attaining a deeper understanding of these matters will create opportunities in the promising area of epigenetic treatments . Identification of novel small molecules that elevate Q9UEF7 expression . The absence of Q9UEF7 ( KL ) from mice causes the development of disorders associated with human aging and decreased longevity , whereas increased expression prolongs lifespan . With age , KL protein levels decrease , and keeping levels consistent may promote healthier aging and be disease-modifying . Using the KL promoter to drive expression of luciferase , we conducted a high-throughput screen to identify compounds that activate KL transcription . Hits were identified as compounds that elevated luciferase expression at least 30 % . Following validation for dose-dependent activation and lack of cytotoxicity , hit compounds were evaluated further in vitro by incubation with opossum kidney and Z310 rat choroid plexus cells , which express KL endogenously . All compounds elevated KL protein compared with control . To determine whether increased protein resulted in an in vitro functional change , we assayed Q9GZV9 ( fibroblast growth factor 23 ) signalling . Compounds G-I augmented P29323 ( extracellular-signal-regulated kinase ) phosphorylation in FGFR ( fibroblast growth factor receptor ) -transfected cells , whereas co-transfection with KL siRNA ( small interfering RNA ) blocked the effect . These compounds will be useful tools to allow insight into the mechanisms of KL regulation . Further optimization will provide pharmacological tools for in vivo studies of KL . P09544 regulates DNA synthesis in mouse granulosa cells through beta-catenin . WNTs are secreted extracellular signaling molecules that transduce their signals by binding to G protein-coupled receptors of the frizzled ( FZD ) family . They control diverse developmental processes , such as cell fate specification , cell proliferation , cell differentiation , and apoptosis . Although WNT signaling has been shown to be essential for development of the ovary , its mechanistic role in folliculogenesis within the adult ovary has not been studied extensively . Therefore , the objective of this study was to investigate the regulation and function of P09544 signaling in mouse granulosa cells . Immunostaining identified P09544 as being expressed in granulosa cells throughout folliculogenesis , but with varying signal strength : in sequential sections , P09544 immunoreactivity was strongest in healthy antral follicles but weak in atretic follicles . Knockdown of P09544 expression using transfected short interfering RNA decreased DNA synthesis in granulosa cells , whereas P09544 overexpression using a recombinant viral vector enhanced it . P09544 knockdown led to accumulation of glycogen synthase kinase-3beta ( P49841 ) in the cytoplasm but reduced the expression of beta-catenin . Conversely , P09544 overexpression reduced the expression of P49841 in the cytoplasm and induced beta-catenin translocation from the membrane into the nucleus . P35222 knockdown also inhibited DNA synthesis in granulosa cells and neutralized the effect of P09544 overexpression . P09544 /beta-catenin signaling had a slight effect on the apoptosis of granulosa cells . Taken together , the data indicate that P09544 regulates beta-catenin localization in granulosa cells , and P09544 /beta-catenin signaling contributes to regulating their proliferation . [ RAC3 nuclear receptor co-activator has a protective role in the apoptosis induced by different stimuli ] . RAC3 belongs to the family of P52701 nuclear receptors coactivators and it is over-expressed in several tumors . We have previously shown that RAC3 is a NF-kappaB coactivator . In this paper , we investigated the role of RAC3 in cell-sensitivity to apoptosis , using H2O2 in the human embryonic kidney cell line ( HEK293 ) , and tumor necrosis factor-related apoptosis inducing ligand ( P50591 ) in a human chronic myeloid leukemia cell line ( K562 ) naturally resistant to P50591 . We observed that the tumoral K562 cells have high levels of RAC3 if compared with the non-tumoral HEK293 cells . The normal or transfected coactivator over-expression inhibits apoptosis through a diminished caspase activity and O95831 nuclear translocation , increased NF-kappaB , AKT and p38 , and decreased P29323 activities . In contrast , inhibition of RAC3 by siRNA induced sensitivity of K562 to P50591 -induced apoptosis . Such results suggest that over-expression of RAC3 contributes to tumor development through molecular mechanisms that do not depend strictly on acetylation and/or steroid hormones , which control cell death . This could be a possible target for future tumor therapies . Candidate gene approach evaluates association between innate immunity genes and breast cancer risk in Korean women . OBJECTIVES : This study was conducted to investigate the role of common variation in innate immunity-related genes as susceptibility factors to breast cancer risk in Korean women . METHODS : Total 1536 single-nucleotide polymorphisms ( SNPs ) in 203 genes were analyzed by Illumina GoldenGate assay in 209 cases and the same numbers of controls . Both SNP and gene-based tests were used to evaluate the association with breast cancer risk . The robustness of results was further evaluated with permutation method , false discovery rate and haplotype analyses . RESULTS : Both SNP and gene-based analyses showed promising associations with breast cancer risk for 17 genes : Q5JRS4 , P12319 , Q15080 , P78357 , P35222 , P03952 , P05107 , O75342 , P20151 , Q9Y616 , Q9Y5K2 , P42226 , P19878 , P22362 , Q9NPY3 , MBP and NOS1 . The most significant association with breast cancer risk was observed for the Q5JRS4 SNP ( rs2494251 , P-value = 1.2 x 10(-4) ) and P12319 SNP ( rs7548864 , P-value = 7.7 x 10(-4) ) . Gene-based permutation and false discovery rate P-values for Q5JRS4 SNP ( rs2494251 ) with breast cancer risk were also significant ( P = 4 x 10(-5) and 0.008 , respectively ) . Haplotype analyses supported these findings that Q5JRS4 and P12319 were most significantly associated with risk for breast cancer ( P = 2 x 10(-4) and 0.004 , respectively ) . CONCLUSION : This study suggests that common genetic variants in the Q5JRS4 and P12319 be strongly associated with breast cancer risk among Korean women . Long-term loading inhibits P27361 /2 phosphorylation and increases P22607 expression in MC3T3-E1 osteoblast cells . Bone tissue homeostasis relies upon the ability of cells to detect and interpret extracellular signals that direct changes in tissue architecture . This study utilized a four-point bending model to create both fluid shear and strain forces ( loading ) during the time-dependent progression of MC3T3-E1 preosteoblasts along the osteogenic lineage . Loading was shown to increase cell number , alkaline phosphatase ( ALP ) activity , collagen synthesis , and the mRNA expression levels of Runx2 , osteocalcin ( OC ) , osteopontin , and cyclo-oxygenase-2 . However , mineralization in these cultures was inhibited , despite an increase in calcium accumulation , suggesting that loading may inhibit mineralization in order to increase matrix deposition . Loading also increased fibroblast growth factor receptor-3 ( P22607 ) expression coincident with an inhibition of P11362 , P22455 , P05230 , and extracellular signal-related kinase ( P29323 )1/2 phosphorylation . To examine whether these loading-induced changes in cell phenotype and FGFR expression could be attributed to the inhibition of P27361 /2 phosphorylation , cells were grown for 25 days in the presence of the Q02750 /2 inhibitor , U0126 . Significant increases in the expression of P22607 , ALP , and OC were observed , as well as the inhibition of P11362 , P22455 , and P05230 . However , U0126 also increased matrix mineralization , demonstrating that inhibition of P27361 /2 phosphorylation can not fully account for the changes observed in response to loading . In conclusion , this study demonstrates that preosteoblasts are mechanoresponsive , and that long-term loading , whilst increasing proliferation and differentiation of preosteoblasts , inhibits matrix mineralization . In addition , the increase in P22607 expression suggests that it may have a role in osteoblast differentiation . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . 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 . 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 . Q9UEF7 gene deficiency causes salt-sensitive hypertension via monocyte chemotactic protein-1/CC chemokine receptor 2-mediated inflammation . Q9UEF7 ( KL ) is a newly discovered aging suppressor gene . In mice , the KL gene extends the lifespan when overexpressed and shortens the lifespan when disrupted . This study investigated if KL deficiency affects BP and salt sensitivity using KL mutant heterozygous ( +/- ) mice and wild-type ( WT ) mice ( 9 weeks of age , 16 mice per group ) . Notably , systolic BP in KL(+/-) mice began to increase at the age of 15 weeks , reached a peak level at the age of 17 weeks , and remained elevated thereafter , whereas systolic BP remained consistent in WT mice . High salt ( HS ) intake further increased BP in KL(+/-) mice but did not affect BP in WT mice . Blockade of CC chemokine receptor 2 ( P41597 ) , involved in monocyte chemotaxis , by a specific P41597 antagonist ( DB05130 ) abolished the HS-induced increase in BP in KL(+/-) mice . Furthermore , HS loading substantially increased the expression of monocyte chemotactic protein-1 and the infiltration of macrophages and T cells in kidneys in KL(+/-) mice , and treatment with DB05130 abolished these effects . Treatment of KL(+/-) mice with DB05130 also attenuated the increased renal expressions of serum glucocorticoid-regulated kinase 1 , thiazide-sensitive NaCl cotransporter , and DB00171 synthase β along with the renal structural damage and functional impairment induced by HS loading . In conclusion , KL deficiency caused salt-sensitive hypertension and renal damage by P41597 -mediated inflammation . Detection of overexpressed P35354 in precancerous lesions of hamster pancreas and lungs by molecular imaging : implications for early diagnosis and prevention . The enzyme cyclooxygenase-2 ( P35354 ) is overexpressed in many cancers , cardiovascular disease , neurodegenerative disorders , and arthritis . Selective inhibitors of P35354 have been developed as therapeutics or preventive agents for these diseases . However , recent reports have revealed a significant increase in cardiovascular mortality in long-term users of the P35354 inhibitors Vioxx and DB00482 , emphasizing the need for noninvasive tests that allow the identification of individuals whose P35354 levels are overexpressed prior to assignment to treatment with these drugs . In this study , we have prepared a radioiodinated analogue of the selective P35354 inhibitor celecoxib , and verified its binding to the P35354 enzyme in vitro . Biodistribution studies in hamsters demonstrated significantly higher levels of radiotracer in animals treated with the tobacco carcinogen NNK in lung , pancreas , and liver . Assessment of P35354 levels by whole-body planar nuclear imaging two hours after injection of the radiotracer was suggestive of a distinct increase in P35354 in the pancreas and liver of a hamster treated for 10 weeks with NNK , in the lungs and liver of a second animal , and in the liver only , in two additional animals from the same treatment group . Immunostains showed selective overexpression of P35354 in pre-neoplastic lesions of the pancreas and lungs in only those animals that showed tracer accumulation in these organs and in the livers of all NNK-treated hamsters . Immunostains for P23219 yielded detectable reactions in the intestinal epithelium but not in pancreas , lungs , or liver , supporting the specificity of the tracer for P35354 . Our data provide proof of principle for the hypothesis that molecular imaging with radiolabeled P35354 inhibitors can be used for the noninvasive monitoring of overexpressed P35354 levels . The genes of the coactivator Q15596 and the corepressor Q9Y618 are primary DB00136 targets . The complex of the receptor for the hormone 1alpha,25-dihydroxyvitamin D(3) ( 1alpha,25(OH)(2)D(3) ) , Vitamin D(3) receptor ( P11473 ) , the retinoid X receptor ( RXR ) and a 1alpha,25(OH)(2)D(3) response element ( VDRE ) is considered to be the molecular switch for nuclear 1alpha,25(OH)(2)D(3) signaling . In the presence of ligand the P11473 -RXR complex interacts with coactivator ( DB01992 ) proteins that in turn contact components of the basal transcriptional machinery resulting in an enhanced transcription of 1alpha,25(OH)(2)D(3) target genes . In the absence of ligand the P11473 remains bound to the DNA and interacts with corepressor ( CoR ) proteins that are involved in gene silencing activity . We treated MCF-7 breast cancer cells with 1alpha,25(OH)(2)D(3) for increasing amounts of time , extracted mRNA and screened by real-time PCR the members of the P52701 DB01992 and NCoR CoR families . We find that of the P52701 coactivators , only Q15596 was responsive to 1alpha,25(OH)(2)D(3) . Similarly Q9Y618 but not NCoR1 gene transcription was sensitive to 1alpha,25(OH)(2)D(3) treatment . In silico analysis revealed that both Q15596 and Q9Y618 promoters have substantial numbers of VDREs compared to the promoters of the other family members . These VDREs are formed by direct repeats of the core binding motif RGKTCA with a three nucleotide spacing ( Q93038 ) . We suggest that some or all of these Q93038 -type VDREs are responsible for the observed responsiveness of Q15596 and Q9Y618 to 1alpha,25(OH)(2)D(3) . Epigenetic therapy of cancer stem and progenitor cells by targeting DNA methylation machineries . Recent advances in stem cell biology have shed light on how normal stem and progenitor cells can evolve to acquire malignant characteristics during tumorigenesis . The cancer counterparts of normal stem and progenitor cells might be occurred through alterations of stem cell fates including an increase in self-renewal capability and a decrease in differentiation and/or apoptosis . This oncogenic evolution of cancer stem and progenitor cells , which often associates with aggressive phenotypes of the tumorigenic cells , is controlled in part by dysregulated epigenetic mechanisms including aberrant DNA methylation leading to abnormal epigenetic memory . Epigenetic therapy by targeting DNA methyltransferases ( P26358 ) 1 , Q9Y6K1 and Q9UBC3 via DB00928 ( Aza ) and 5-Aza-2'-deoxycytidine ( Aza-dC ) has proved to be successful toward treatment of hematologic neoplasms especially for patients with myelodysplastic syndrome . In this review , I summarize the current knowledge of mechanisms underlying the inhibition of DNA methylation by Aza and Aza-dC , and of their apoptotic- and differentiation-inducing effects on cancer stem and progenitor cells in leukemia , medulloblastoma , glioblastoma , neuroblastoma , prostate cancer , pancreatic cancer and testicular germ cell tumors . Since cancer stem and progenitor cells are implicated in cancer aggressiveness such as tumor formation , progression , metastasis and recurrence , I propose that effective therapeutic strategies might be achieved through eradication of cancer stem and progenitor cells by targeting the DNA methylation machineries to interfere their " malignant memory " . 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 . Effect of increased glycogen synthase kinase-3 activity upon the maturation of the amyloid precursor protein in transfected cells . Aberrant control of protein phosphorylation is an important feature in Alzheimer 's disease pathology . The action of glycogen synthase kinase-3 beta ( P49841 ) on the maturation and phosphorylation of an amyloid precursor protein-reporter construct ( P05067 -REP ) was studied in transfected COS-7 cells . Elevation of P49841 activity by enzyme over-expression resulted in an increase in the level of mature forms of co-expressed P05067 -REP . This effect was not associated with an increased level of P05067 -REP phosphorylation at Thr743 , an in vitro P49841 phosphorylation site . These findings suggest that P49841 activity may indirectly increase cellular maturation of P05067 , which may subsequently result in altered production of beta-amyloid protein . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . 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 . Effect of active smoking on the human bronchial epithelium transcriptome . BACKGROUND : Lung cancer is the most common cause of cancer-related deaths . Tobacco smoke exposure is the strongest aetiological factor associated with lung cancer . In this study , using serial analysis of gene expression ( Q9NXZ1 ) , we comprehensively examined the effect of active smoking by comparing the transcriptomes of clinical specimens obtained from current , former and never smokers , and identified genes showing both reversible and irreversible expression changes upon smoking cessation . RESULTS : Twenty-four Q9NXZ1 profiles of the bronchial epithelium of eight current , twelve former and four never smokers were generated and analyzed . In total , 3,111,471 Q9NXZ1 tags representing over 110 thousand potentially unique transcripts were generated , comprising the largest human Q9NXZ1 study to date . We identified 1,733 constitutively expressed genes in current , former and never smoker transcriptomes . We have also identified both reversible and irreversible gene expression changes upon cessation of smoking ; reversible changes were frequently associated with either xenobiotic metabolism , nucleotide metabolism or mucus secretion . Increased expression of Q07654 , O75952 , and Q5MY95 were found to be reversible upon smoking cessation . Expression of P49841 , which regulates P35354 expression , was irreversibly decreased . P98088 expression was only partially reversed . Validation of select genes was performed using quantitative RT-PCR on a secondary cohort of nine current smokers , seven former smokers and six never smokers . CONCLUSION : Expression levels of some of the genes related to tobacco smoking return to levels similar to never smokers upon cessation of smoking , while expression of others appears to be permanently altered despite prolonged smoking cessation . These irreversible changes may account for the persistent lung cancer risk despite smoking cessation .

MH_train_1478

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MH_train_1478

interacts_with DB01267?

multiple_choice

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 . 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 . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Genotoxicity and induction of DNA damage responsive genes by food-borne heterocyclic aromatic amines in human hepatoma HepG2 cells . Heterocyclic aromatic amines ( HAAs ) are potential human carcinogens formed in well-done meats and fish . The most abundant are 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline ( MeIQx ) , 2-Amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinoxaline ( 4,8-DiMeIQx ) and 2-Amino-3-methyl-3H-imidazo[4,5-f]quinoline ( IQ ) . HAAs exert genotoxic activity after metabolic transformation by CYP1A enzymes , that is well characterized , however the genomic and intervening responses are not well explored . We have examined cellular and genomic responses of human hepatoma HepG2 cells after 24h exposure to HAAs . Comet assay revealed increase in formation of DNA strand breaks by PhIP , MeIQx and IQ but not 4,8-DiMeIQx , whereas increased formation of micronuclei was not observed . The four HAAs up-regulated expression of genes encoding metabolic enzymes P04798 , P05177 and P22309 and expression of P04637 and its downstream regulated genes P38936 , GADD45α and Q07812 . Consistent with the up-regulation of P38936 and GADD45α the cell-cycle analysis showed arrest in S-phase by PhIP and IQ , and in P55008 -phase by 4,8-DiMeIQx and MeIQx . The results indicate that upon exposure to HAAs the cells respond with the cell-cycle arrest , which enables cells to repair the damage or eliminate them by apoptosis . However , elevated expression of P10415 and down-regulation of Q07812 may indicate that HAAs could suppress apoptosis meaning higher probability of damaged cells to survive and mutate . 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 . 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 . Glucocorticoids inhibit tetrahydrobiopterin-dependent endothelial function . DB00360 ( BH4 ) acts as an important co-factor for endothelial nitric oxide synthase ( P29474 ) . Glucocorticoids have been shown to inhibit expression of the rate-limiting enzyme for tetrahydrobiopterin synthesis , GTP cyclohydrolase , in other cell types . We hypothesized that endothelium-dependent vasodilator responses would be blunted in rats made hypertensive with dexamethasone . Further , we hypothesized that treatment of rat vascular segments with dexamethasone would result in attenuation of endothelial function accompanied by decreased GTP cyclohydrolase expression . We report that endothelium-dependent relaxation responses to the calcium ionophore A23187 are reduced in aortic rings from dexamethasone-hypertensive rats compared with sham values . Dexamethasone incubation abolishes contraction to Nomega-nitro-L-arginine ( DB04223 , 10(-5) M ) in endothelium-intact aortic rings , and inhibits expression of GTP cyclohydrolase . We conclude that inhibition of BH4 synthesis by glucocorticoid regulation of GTP cyclohydrolase expression may contribute to reduced endothelium-dependent vasodilation characteristic of glucocorticoid-induced hypertension . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . Distinct pharmacological properties of second generation HDAC inhibitors with the benzamide or hydroxamate head group . Advanced second generation inhibitors of histone deacetylases ( HDAC ) are currently used in clinical development . This study aimed at comparing the pharmacological properties of selected second generation HDAC inhibitors with the hydroxamate and benzamide head group , namely DB02546 , LAQ824/LBH589 , CI994 , MS275 and DB05651 . In biochemical assays using recombinant Q13547 , 3 , 6 and 8 isoenzymes , DB02546 and LAQ824/LBH589 behave as quite unselective HDAC inhibitors . In contrast , the benzamides CI994 , MS275 and DB05651 are more selective , potent inhibitors of at least Q13547 and O15379 . All HDAC inhibitors induce histone H3 hyperacetylation , correlating with inhibition of proliferation , induction of cell differentiation and apoptosis . A broad cytotoxicity is seen across cell lines from different tumor entities with LAQ824/LBH589 being the most potent agents . The apoptosis inducing activity is evident in arrested and proliferating RKO colon cancer cells with inducible , heterologous P38936 (waf1) expression , indicative for a cell-cycle independent mode-of-action . Differentiation of MDA-MB468 breast cancer cells is induced by benzamide and hydroxamate analogs . The reversibility of drug action was evaluated by pulse treatment of A549 lung cancer cells . Whereas paclitaxel induced irreversible cell cycle alterations already after 6 hr treatment , HDAC inhibitor action was retarded and irreversible after > 16 hr treatment . Interestingly , pulse treatment was equally effective as continous treatment . Finally , the efficacy of LAQ824 , DB02546 and MS275 in A549 nude mice xenografts was comparable to that of paclitaxel at well tolerated doses . We conclude that despite a different HDAC isoenzyme inhibition profile , hydroxamate and benzamide analogs as studied display similar cellular profiles . 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 . ZNF804a regulates expression of the schizophrenia-associated genes Q9NQE7 , P21964 , Q07343 , and P14416 . ZNF804a was identified by a genome-wide association study ( GWAS ) in which a single nucleotide polymorphism ( SNP rs1344706 ) in ZNF804a reached genome-wide statistical significance for association with a combined diagnosis of schizophrenia ( SZ ) and bipolar disorder . Although the molecular function of ZNF804a is unknown , the amino acid sequence is predicted to contain a C2H2-type zinc-finger domain and suggests ZNF804a plays a role in DNA binding and transcription . Here , we confirm that ZNF804a directly contributes to transcriptional control by regulating the expression of several SZ associated genes and directly interacts with chromatin proximal to the promoter regions of Q9NQE7 and P21964 , the two genes we find upregulated by ZNF804a . Using immunochemistry we establish that ZNF804a is localized to the nucleus of rat neural progenitor cells in culture and in vivo . We demonstrate that expression of ZNF804a results in a significant increase in transcript levels of Q9NQE7 and P21964 , relative to GFP transfected controls , and a statistically significant decrease in transcript levels of Q07343 and P14416 . Furthermore , we show using chromatin immunoprecipitation assays ( ChIP ) that both epitope-tagged and endogenous ZNF804a directly interacts with the promoter regions of Q9NQE7 and P21964 , suggesting a direct upregulation of transcription by ZNF804a on the expression of these genes . These results are the first to confirm that ZNF804a regulates transcription levels of four SZ associated genes , and binds to chromatin proximal to promoters of two SZ genes . These results suggest a model where ZNF804a may modulate a transcriptional network of SZ associated genes . 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 . P14416 signaling dynamics of dopamine D2-neurotensin 1 receptor heteromers . Biochemical , histochemical and coimmunoprecipitation experiments have indicated the existence of antagonistic dopamine D2 ( D2R ) and neurotensin 1 ( NTS1R ) receptor-receptor interactions in the dorsal and ventral striatum indicating a potential role of these receptor-receptor interactions in Parkinson 's disease and schizophrenia . By means of Bioluminiscence Resonance energy transfer ( BRET(2) ) evidence has for the first time been obtained in the current study for the existence of both D2LR/NTS1R and D2SR/NTS1R heteromers in living HEK293T cells . Through confocal laser microscopy the NTS1R(GFP2) and D2R(YFP) were also shown to be colocated in the plasma membrane of these cells . A bioinformatic analysis suggests the existence of a basic set of three homology protriplets ( TVM , DLL and/or LRA ) in the two participating receptors which may contribute to the formation of the D2R/NTS1R heteromers by participating in guide-clasp interactions in the receptor interface . The CREB reporter gene assay indicated that the neurotensin receptor agonist JMV 449 markedly reduced the potency of the D2R like agonist quinpirole to inhibit the forskolin induced increase of the CREB signal . In contrast , the neurotensin agonist was found to markedly increase the quinpirole potency to activate the MAPK pathway as also studied with luciferase reporter gene assay measuring the degree of SRE activity as well as with P27361 /2 phosphorylation assays . These dynamic changes in D2R signaling produced by the neurotensin receptor agonist may involve antagonistic allosteric receptor-receptor interactions in the D2LR-NTS1R heteromers at the plasma membrane level ( CREB pathway ) and synergistic interactions in PKC activation at the cytoplasmatic level ( MAPK pathway ) . DB00898 blocks leukocyte recruitment and endothelial cell activation in vivo and in vitro . Immune system impairment and increased susceptibility to infection among alcohol abusers is a significant but not well-understood problem . We hypothesized that acute ethanol administration would inhibit leukocyte recruitment and endothelial cell activation during inflammation and infection . Using LPS and carrageenan air pouch models in mice , we found that physiological concentrations of ethanol ( 1-5 g/kg ) significantly blocked leukocyte recruitment ( 50-90 % ) . Because endothelial cell activation and immune cell-endothelial cell interactions are critical regulators of leukocyte recruitment , we analyzed the effect of acute ethanol exposure on endothelial cell activation in vivo using the localized Shwartzman reaction model . In this model , ethanol markedly suppressed leukocyte accumulation and endothelial cell adhesion molecule expression in a dose-dependent manner . Finally , we examined the direct effects of ethanol on endothelial cell activation and leukocyte-endothelial cell interactions in vitro . DB00898 , at concentrations within the range found in human blood after acute exposure and below the levels that induce cytotoxicity ( 0.1-0.5 % ) , did not induce endothelial cell activation , but significantly inhibited P01375 -mediated endothelial cell activation , as measured by adhesion molecule ( P16581 , P05362 , P19320 ) expression and chemokine ( P10145 , P13500 , RANTES ) production and leukocyte adhesion in vitro . Studies exploring the potential mechanism by which ethanol suppresses endothelial cell activation revealed that ethanol blocked NF-kappaB nuclear entry in an P25963 -dependent manner . These findings support the hypothesis that acute ethanol overexposure may increase the risk of infection and inhibit the host inflammatory response , in part , by blocking endothelial cell activation and subsequent immune cell-endothelial cell interactions required for efficient immune cell recruitment . 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 . 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 . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 .

MH_train_1479

MH_train_1479

MH_train_1479

interacts_with DB01211?

multiple_choice

Pharmacologic suppression of hepatic O95477 activity in mice reduces high-density lipoprotein cholesterol levels but promotes reverse cholesterol transport . BACKGROUND : The role of hepatic O95477 ( O95477 ) in maintaining plasma high density lipoprotein cholesterol ( HDL-C ) levels is well established , but its role in reverse cholesterol transport ( RCT ) is unclear . DB01599 is a compound that reduces HDL-C levels but also reduces atherosclerosis in animal models and xanthomas in humans . The aim of the present study was to test the hypothesis that probucol inhibits hepatic O95477 activity , thereby reducing HDL-C levels but promoting RCT from macrophages . METHODS AND RESULTS : Wild-type ( WT ) C57BL/6 mice and scavenger receptor class B type I ( Q8WTV0 ) knockout mice were fed a chow diet containing 0.5 % probucol or normal chow for 2 weeks . In WT mice , probucol , despite decreasing HDL-C by > 80 % , effectively maintained macrophage RCT . In Q8WTV0 knockout mice , probucol also substantially reduced HDL-C but significantly increased macrophage RCT . Furthermore , probucol significantly enhanced the excretion of HDL-derived cholesterol into feces in both WT and Q8WTV0 knockout mice . DB01599 inhibited O95477 -dependent cholesterol efflux from mouse primary hepatocytes , and this effect was shown to be responsible for the effect of probucol on increasing the fecal excretion of HDL-derived cholesterol in vivo . CONCLUSIONS : We demonstrate that pharmacological inhibition of hepatic O95477 activity with probucol reduced HDL-C levels but promoted RCT through diversion of HDL-derived cholesterol from efflux back into plasma instead to excretion in the bile . These results explain the beneficial effects of probucol on atherosclerosis and xanthomas despite its HDL-lowering effects and suggest that inactivation of hepatic O95477 leads to increased RCT despite reducing plasma HDL-C levels . PDE10 inhibition increases P42261 and CREB phosphorylation and improves spatial and recognition memories in a Huntington 's disease mouse model . Huntington 's disease ( HD ) causes motor disturbances , preceded by cognitive impairment , in patients and mouse models . We showed that increased hippocampal DB02527 -dependent protein kinase ( PKA ) signaling disrupts recognition and spatial memories in R6 HD mouse models . However , unchanged levels of hippocampal phosphorylated ( p ) DB02527 -responsive element-binding protein ( CREB ) suggested unaltered nuclear PKA activity in R6 mice . Here , we extend this finding by showing that nuclear pPKA catalytic subunit ( Thr197 ) and pPKA substrate levels were unaltered in the hippocampus of R6/1 mice . Phosphodiesterases ( PDEs ) play an important role in the regulation of PKA activity . Q9Y233 , a DB02527 /cGMP dual-substrate PDE , was reported to be restricted to the nuclear region in nonstriatal neurons . Using cell fractionation we confirmed that Q9Y233 was enriched in nuclear fractions , both in wild-type and R6/1 mice hippocampus , without differences in its levels or intracellular distribution between genotypes . We next investigated whether inhibition of PDE10 with papaverine could improve cognitive function in HD mice . DB01113 treatment improved spatial and object recognition memories in R6/1 mice , and significantly increased pGluA1 and pCREB levels in R6/1 mice hippocampus . DB01113 likely acted through the activation of the PKA pathway as the phosphorylation level of distinct cGMP-dependent kinase ( cGK ) substrates was not modified in either genotype . Moreover , hippocampal DB02527 , but not cGMP , levels were increased after acute papaverine injection . Our results show that inhibition of PDE10 improves cognition in R6 mice , at least in part through increased P42261 and CREB phosphorylation . Thus , PDE10 might be a good therapeutic target to improve cognitive impairment in HD . [ Transport of mucoid mucus in healthy individuals and patients with chronic obstructive pulmonary disease and bronchiectasis ] . OBJECTIVE : To characterise and compare the in vitro transport properties of respiratory mucoid secretion in individuals with no lung disease and in stable patients with chronic obstructive pulmonary disease ( P48444 ) and bronchiectasis . METHODOLOGY : Samples of mucus were collected , from 21 volunteers presenting no lung disease who had undergone surgery , from 10 patients presenting chronic P48444 , and from 16 patients with bronchiectasis . Mucociliary transport ( Q8IVS2 ) , transport by cough ( DB00919 ) , and contact angle ( P62158 ) were evaluated . RESULTS : Q8IVS2 was found to be greater in healthy individuals ( 1.0±0.19 ) than in P48444 ( 0.91±0.17 ) and bronchiectasis ( 0.76±0.23 ) patients ( p < 0.05 ) , whereas DB00919 was greater in P48444 patients ( 16.31±7.35 cm ) than in patients with bronchiectasis ( 12.16±6.64 cm ) and healthy individuals ( 10.50±25.8 cm ) ( p < 0.05 ) . No significant differences were observed between the groups regarding P62158 . CONCLUSION : Mucus from healthy individuals allows better mucociliary transport compared to that from patients with lung diseases . However , the mucus from P48444 patients allows a better transport by coughing , demonstrating that these individuals have adapted to a defence mechanism compared to patients with bronchiectasis , who have impairment in their ciliary and cough transport mechanisms . Effects of ozone exposure mediated by BEAS-2B cells on T cells activation : a possible link between environment and asthma . OBJECTIVE : To explore the possible link between ozone and asthma through analyzing Th1/Th2 differentiation of T cells following incubation with conditioned medium from the BEAS-2B cells exposed to ozone in vitro . METHOD : Bronchial epithelial cell line , BEAS-2B , was cultured using an air-liquid interface culture system in a CO2 incubator and exposed to 0 or 0.16 or 0.25 mg/m3 of ozone for 8 h . The amounts of IL-1β , P05231 and RANTES in the cell supernatant were detected . The cell culture supernatants were collected and used as conditioned medium in the next experiment . T cells from children recruited were incubated with conditioned medium for 12 h . Activation rate of Q07108 and Th1/Th2/Th17 differentiation were analyzed . RESULTS : BEAS-2B cells exposed to different ozone concentrations showed morphological changes . Cells exposed to 0.16 and 0.25 mg/m3 ozone produced higher amounts of IL-1β , P05231 and RANTES than that in the control group . Children with allergic asthma had upregulated expression of genes related with asthma , including P13500 , CCR4 , P19875 , Q9Y271 , Q99665 , Q14627 , Q13478 , P01584 , P10145 , P25025 and O75888 . Q07108 expression in T cells was significantly elevated irrespective of ozone exposure in children with allergic asthma . Following ozone exposure , in asthmatic children group , expression levels of cytokines of Th1 cells were collectively higher than those from Th2 cells . Ozone-exposed conditioned media could slightly increase all the Th1 , Th2 and Th17 cytokines in T cells from allergic asthmatic children . CONCLUSIONS : Our results suggested that Th1 cells activation might be predominant over Th2 activation upon ozone exposure in asthmatic children , which might help to clarify the mechanisms of asthma related to environmental factors like ozone . 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 . 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 . [ 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 . 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 . 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 . DB08915 , a balanced PPARα/γ agonist , has no clinically relevant pharmacokinetic interaction with high-dose atorvastatin or rosuvastatin . BACKGROUND : DB08915 , a dual Q07869 -α/γ agonist , combines the lipid benefits of fibrates and the insulin-sensitizing benefits of thiazolidinediones . OBJECTIVE : To investigate the pharmacokinetic effects of co-administration of atorvastatin or rosuvastatin with aleglitazar . RESEARCH DESIGN AND METHODS : In a two-cohort , open-label , randomised , three-period crossover study , 44 healthy subjects received once-daily oral doses of aleglitazar 300 μg , statin ( atorvastatin 80 mg or rosuvastatin 40 mg ) and aleglitazar co-administered with each statin for 7 days . Plasma concentrations of each drug were measured and pharmacokinetic parameters determined on day 7 in each period . MAIN OUTCOME MEASURES : Peak observed plasma concentration ( C(max) ) and total exposures ( AUC ( 0 - 24 ) ) of aleglitazar , atorvastatin and rosuvastatin . RESULTS : C(max) and AUC ( 0 - 24 ) to aleglitazar were similar , whether administered alone or in combination with a statin . Total exposure to either statin was unaffected by co-administration with aleglitazar . C(max) treatment ratios for both statins exceeded the conventional no-effect boundary ( 1.25 ) when administered with aleglitazar . CONCLUSIONS : Co-administration of aleglitazar with a statin does not alter the pharmacokinetic profile of either drug . 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 . 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 . 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 . 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 . Ca2+/ P62158 -dependent protein kinase kinase beta is regulated by multisite phosphorylation . Ca(2+)/calmodulin-dependent protein kinase kinase β ( CaMKKβ ) is a serine/threonine-directed kinase that is activated following increases in intracellular Ca(2+) . CaMKKβ activates Ca(2+)/calmodulin-dependent protein kinase I , Ca(2+)/calmodulin-dependent protein kinase IV , and the AMP-dependent protein kinase in a number of physiological pathways , including learning and memory formation , neuronal differentiation , and regulation of energy balance . Here , we report the novel regulation of CaMKKβ activity by multisite phosphorylation . We identify three phosphorylation sites in the N terminus of CaMKKβ , which regulate its Ca(2+)/calmodulin-independent autonomous activity . We then identify the kinases responsible for these phosphorylations as cyclin-dependent kinase 5 ( Q00535 ) and glycogen synthase kinase 3 ( GSK3 ) . In addition to regulation of autonomous activity , we find that phosphorylation of CaMKKβ regulates its half-life . We find that cellular levels of CaMKKβ correlate with Q00535 activity and are regulated developmentally in neurons . Finally , we demonstrate that appropriate phosphorylation of CaMKKβ is critical for its role in neurite development . These results reveal a novel regulatory mechanism for CaMKKβ-dependent signaling cascades . Cav1.2 L-type Ca²⁺ channels mediate cocaine-induced P42261 trafficking in the nucleus accumbens , a long-term adaptation dependent on ventral tegmental area Ca(v)1.3 channels . AMPA receptor ( AMPAR ) plasticity at glutamatergic synapses in the mesoaccumbal dopaminergic pathway has been implicated in persistent cocaine-induced behavioral responses ; however , the precise mechanism underlying these changes remains unknown . Utilizing cocaine psychomotor sensitization , we have examined phosphorylation of P42261 at key residues serine 845 ( S845 ) and S831 , as well as P42261 cell surface levels in the nucleus accumbens ( NAc ) of cocaine-preexposed mice and the role of brain-specific Ca(v)1.2 and Ca(v)1.3 L-type Ca²⁺ channels ( LTCCs ) , therein . We found higher basal levels of S845 phospho- P42261 ( P- P42261 ) and cell surface P42261 in the NAc following protracted withdrawal from cocaine exposure , changes that occur independently of LTCCs . In contrast , we found that a cocaine challenge that elicits expression of the cocaine-sensitized response increases S831 P- P42261 that further increases surface P42261 beyond the higher basal levels . Intra-NAc pharmacological manipulations indicate that the Ca(v)1.2-activated P62158 kinase II ( CaMKII ) mediates cocaine-induced increase in S831 P- P42261 and that both Ca(v)1.2-activated CaMKII and extracellular signal-regulated kinase 2 ( P28482 ) mediate the increase in P42261 cell surface levels specific to the sensitized response . Experiments using adenoassociated viral vectors expressing Ca(v)1.3 and P28482 siRNA further indicate that recruitment of the Ca(v)1.2 pathway in the NAc is dependent on ventral tegmental area Ca(v)1.3 LTCCs and P28482 . Together , these results identify candidate pathways that mediate cocaine-induced AMPAR plasticity in the NAc and provide a mechanism linking LTCCs and P42261 plasticity to cocaine-induced persistent behavioral changes . How corticosteroids control inflammation : Quintiles Prize Lecture 2005 . Corticosteroids are the most effective anti-inflammatory therapy for many chronic inflammatory diseases , such as asthma but are relatively ineffective in other diseases such as chronic obstructive pulmonary disease ( P48444 ) . Chronic inflammation is characterised by the increased expression of multiple inflammatory genes that are regulated by proinflammatory transcription factors , such as nuclear factor-kappaB and activator protein-1 , that bind to and activate coactivator molecules , which then acetylate core histones to switch on gene transcription . Corticosteroids suppress the multiple inflammatory genes that are activated in chronic inflammatory diseases , such as asthma , mainly by reversing histone acetylation of activated inflammatory genes through binding of liganded glucocorticoid receptors ( GR ) to coactivators and recruitment of histone deacetylase-2 ( Q92769 ) to the activated transcription complex . At higher concentrations of corticosteroids GR homodimers also interact with DNA recognition sites to active transcription of anti-inflammatory genes and to inhibit transcription of several genes linked to corticosteroid side effects . In patients with P48444 and severe asthma and in asthmatic patients who smoke Q92769 is markedly reduced in activity and expression as a result of oxidative/nitrative stress so that inflammation becomes resistant to the anti-inflammatory actions of corticosteroids . DB00277 , by activating HDAC , may reverse this corticosteroid resistance . This research may lead to the development of novel anti-inflammatory approaches to manage severe inflammatory diseases . Traumatic brain injury-induced acute gene expression changes in rat cerebral cortex identified by GeneChip analysis . Proper CNS function depends on concerted expression of thousands of genes in a controlled and timely manner . Traumatic brain injury ( TBI ) in mammals results in neuronal death and neurological dysfunction , which might be mediated by altered expression of several genes . By employing a CNS-specific GeneChip and real-time polymerase chain reaction ( PCR ) , the present study analyzed the gene expression changes in adult rat cerebral cortex in the first 24 hr after a controlled cortical impact injury . Many functional families of genes not previously implicated in TBI-induced brain damage are altered in the injured cortex . These include up-regulated transcription factors ( O14543 , O60674 , P35610 -3 , Q03060 , P10914 , SMN , silencer factor-B , ANIA-3 , ANIA-4 , and DB09106 -1 ) and signal transduction pathways ( cpg21 , Narp , and P09455 ) and down-regulated transmitter release mechanisms ( CITRON , synaptojanin II , ras-related rab3 , neurexin-1beta , and SNAP25A and -B ) , kinases ( IP-3-kinase , Pak1 , Ca(2+)/ P62158 -dependent protein kinases ) , and ion channels ( K(+) channels TWIK , RK5 , X62839 , and Na(+) channel I ) . In addition , several genes previously shown to play a role in TBI pathophysiology , including proinflammatory genes , proapoptotic genes , heat shock proteins , immediate early genes , neuropeptides , and glutamate receptor subunits , were also observed to be altered in the injured cortex . Real-time PCR analysis confirmed the GeneChip data for many of these transcripts . The novel physiologically relevant gene expression changes observed here might explain some of the molecular mechanisms of TBI-induced neuronal damage . 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 . Inhibition of angiogenesis by HC·HA , a complex of hyaluronan and the heavy chain of inter-α-inhibitor , purified from human amniotic membrane . PURPOSE : To determine whether antiangiogenic action of the amniotic membrane ( AM ) can be mediated by HC·HA , a covalent complex of hyaluronan ( HA ) and the heavy chain ( HC ) of inter-α-inhibitor , purified from AM soluble extract . METHODS : HC·HA action on viability , proliferation , attachment , death , migration , and differentiation of human umbilical vein endothelial cells ( HUVECs ) and neovascularization in chicken chorioallantoic membrane ( P62158 ) was examined by MTT assay , BrdU labeling , cell proliferation assay , cell death detection ELISA , transwell assay , tube formation assay , and P62158 assay . RESULTS : HC·HA suppressed HUVEC viability more significantly than HA and AM stromal extract , and such suppression was not mediated by P16070 . HC·HA also caused HUVECs to become small and rounded , with a decrease in spreading and filamentous actin . Without promoting cell detachment or death , HC·HA dose dependently inhibited proliferation ( IC(50) , 2.3 μg/mL ) and was 100-fold more potent than HA . Migration triggered by P15692 and tube formation was also significantly inhibited by HC·HA . Purified HC·HA did not contain P36955 and P07996 -1 but did contain P08833 and platelet factor 4 while significantly suppressing neovascularization in P62158 . CONCLUSIONS : The antiangiogenic activity of HC·HA might explain why AM is developmentally avascular and how AM might exert an antiangiogenic action when transplanted to the ocular surface , and it might indicate a potential therapeutic effect of HC·HA in diseases manifesting pathogenic angiogenesis . Roles of P08833 and platelet factor 4 in HC·HA antiangiogenic action warrant further investigation . Q16236 plays a protective role in diabetic retinopathy in mice . AIMS/HYPOTHESIS : Although much is known about the pathophysiological processes contributing to diabetic retinopathy ( DR ) , the role of protective pathways has received less attention . The transcription factor nuclear factor erythroid-2-related factor 2 ( also known as Q16236 or Q16236 ) is an important regulator of oxidative stress and also has anti-inflammatory effects . The objective of this study was to explore the potential role of Q16236 as a protective mechanism in DR . METHODS : Retinal expression of Q16236 was investigated in human donor and mouse eyes by immunohistochemistry . The effect of Q16236 modulation on oxidative stress was studied in the human Müller cell line Q9NP71 -M1 . Non-diabetic and streptozotocin-induced diabetic wild-type and Nrf2 knockout mice were evaluated for multiple DR endpoints . RESULTS : Q16236 was expressed prominently in Müller glial cells and astrocytes in both human and mouse retinas . In cultured Q9NP71 -M1 cells , Q16236 inhibition significantly decreased antioxidant gene expression and exacerbated tert-butyl hydroperoxide- and hydrogen peroxide-induced oxidative stress . Q16236 activation strongly increased Q16236 target gene expression and suppressed oxidant-induced reactive oxygen species . Diabetic mice exhibited retinal Q16236 activation , indicated by nuclear translocation . Superoxide levels were significantly increased by diabetes in Nrf2 knockout mice as compared with wild-type mice . Diabetic Nrf2 knockout mice exhibited a reduction in retinal glutathione and an increase in P01375 -α protein compared with wild-type mice . Nrf2 knockout mice exhibited early onset of blood-retina barrier dysfunction and exacerbation of neuronal dysfunction in diabetes . CONCLUSIONS/INTERPRETATION : These results indicate that Q16236 is an important protective factor regulating the progression of DR and suggest enhancement of the Q16236 pathway as a potential therapeutic strategy .

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interacts_with DB00227?

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Cigarette smoke-induced kinin B1 receptor promotes NADPH oxidase activity in cultured human alveolar epithelial cells . Pulmonary inflammation is an important pathological feature of tobacco smoke-related lung diseases . Kinin B1 receptor ( P46663 ) is up-regulated in the rat trachea chronically exposed to cigarette-smoke . This study aimed at determining ( 1 ) whether exposure to total particulate matter of the cigarette smoke ( DB00273 ) can induce P46663 in human alveolar epithelial A549 cells , ( 2 ) the mechanism of P46663 induction , ( 3 ) the functionality of de novo synthesized P46663 , and ( 4 ) the role of P46663 in DB00273 -induced increase of superoxide anion ( O₂(●⁻) ) level . Results show that A549 cells exposed to 10 μg/ml DB00273 increased O₂(●⁻) level along with P46663 ( protein and mRNA ) and IL-1β mRNA . In contrast , P30411 and P01375 -α mRNA were not affected by DB00273 . The increasing effect of DB00273 on O₂(●⁻) level was not significantly affected by the P46663 antagonist SSR240612 . DB00273 -increased P46663 mRNA was prevented by co-treatments with N-acetyl-l-cysteine ( potent antioxidant ) , diphenyleneiodonium ( NADPH oxidase inhibitor ) , IL-1Ra ( interleukin-1R antagonist ) and SN-50 ( specific inhibitor of NF-kB activation ) but not by pentoxifylline ( P01375 -α release inhibitor ) , indomethacin and niflumic acid ( P23219 and -2 inhibitors ) . Stimulation of P46663 with a selective agonist ( des-Arg⁹-BK , 10 μM ; 30 min ) increased O₂(●⁻)production which was prevented by apocynin and diphenyleneiodonium ( NADPH oxidase inhibitors ) . Data suggest that the increased expression of P46663 by DB00273 in A549 cells is mediated by oxidative stress , IL-1β and NF-kB but not by cyclooxygenases or P01375 -α . The amplification of O₂(●⁻) levels via the activation of P46663 -NADPH oxidase may exacerbate pulmonary inflammation and contribute to the chronicity of tobacco smoke-related lung diseases . DB09152 -bisphosphonates block retinoblastoma phosphorylation and cell growth by inhibiting the cholesterol biosynthetic pathway in a keratinocyte model for esophageal irritation . The surprising discovery that nitrogen-containing bisphosphonates ( N-BPs ) act via inhibition of the mevalonate-to-cholesterol pathway raised the possibility that esophageal irritation by N-BPs is mechanism-based . We used normal human epidermal keratinocytes ( NHEKs ) to model N-BP effects on stratified squamous epithelium of the esophagus . The N-BPs alendronate and risedronate inhibited NHEK growth in a dose-dependent manner without inducing apoptosis . N-BPs ( 30 microM ) caused accumulation of cells in S phase and increased binucleation ( inhibited cytokinesis ) . Consistent with N-BP inhibition of isoprenylation , geranylgeraniol or farnesol prevented accumulation in S phase . Binucleation was also induced by the P04035 inhibitor lovastatin and by the squalene synthase inhibitor zaragozic acid A and was prevented by adding low-density lipoprotein . At 300 microM , N-BPs reduced expression of cyclin-dependent kinase ( cdk ) 2 and cdk4 and enhanced expression of P38936 (waf1) and p27(kip1) and their binding to cdks with corollary hypophosphorylation of retinoblastoma . DB00227 and zaragozic acid A produced similar effects , except that P38936 (waf1) expression and binding to cdks was not induced . Growth inhibition , but not binucleation , was also caused by the geranylgeranyl transferase I inhibitor , GGTI-298 , which also enhanced cdk2 and cdk4 association with p27(kip1) . These findings are consistent with suppression of epithelial cell growth by N-BPs via inhibition of the mevalonate pathway and the consequent reduction in cholesterol synthesis , which blocks cytokinesis , and in geranylgeranylation , which interferes with progression through the cell cycle . 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 . 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 . Attenuation of the progression of adjuvant-induced arthritis by 3-aminobenzamide treatment . Rheumatoid arthritis ( RA ) is a disease that is still insufficiently controlled by current treatments . Poly(ADP-ribose) polymerase ( PARP ) inhibitors ameliorate immune-mediated diseases in several experimental models , including RA , colitis , experimental autoimmune encephalomyelitis and allergy . Together these findings showed that ADP-ribosylating enzymes , in particular P09874 , play a pivotal role in the regulation of immune responses and may represent a noble target for new therapeutic approaches in immune-mediated diseases . The effect of 3-aminobenzamide ( 3-AB ) , an inhibitor of poly(ADP-ribose) synthetase activity , was evaluated in a mouse model of adjuvant-induced arthritis ( AIA ) on pro-inflammatory cytokines , adhesion molecules , inflammatory mediators and chemokine production/expression in serum and knee joint . Histopathological examination was also done on joint section . Our data demonstrates that 3-AB , 10mg/kg , intraperitoneally ( i.p. ) significantly reduces pro-inflammatory cytokine ( Q16552 , P01375 -α and P60568 ) and chemokine ( P13500 and MIP-2 ) production/expression , accompanied by amelioration of the disease as indicated by reduced paw swelling and arthritic scores and was associated with a significant reduction of P19320 and P05362 expression in the knee joint . Moreover , the expression of inflammatory mediators ( P35228 , P35354 , P08253 , P14780 ) and joint histological inflammatory damage was also markedly decreased . The results of this study suggest that P09874 inhibitor may play a role in the inflammatory arthritic process after administration of 3-AB may be a beneficial therapeutic approach . P04271 /RAGE-dependent activation of microglia via NF-kappaB and AP-1 Co-regulation of P35354 expression by P04271 , IL-1beta and P01375 . Extracellular P04271 is known to affect astrocytic , neuronal and microglial activities , with different effects depending on its concentration . Whereas at relatively low concentrations P04271 exerts trophic effects on neurons and astrocytes , at relatively high concentrations the protein causes neuronal apoptosis and activates astrocytes and microglia , thus potentially representing an endogenous factor implicated in neuroinflammation . We have reported that RAGE ligation by P04271 in BV-2 microglia results in the upregulation of expression of the pro-inflammatory cyclo-oxygenase 2 ( P35354 ) via parallel Ras-Cdc42-Rac1-dependent activation of c-Jun NH(2) terminal protein kinase ( JNK ) and Ras-Rac1-dependent stimulation of NF-kappaB transcriptional activity . We show here that : ( 1 ) P04271 also stimulates AP-1 transcriptional activity in microglia via RAGE-dependent activation of JNK ; ( 2 ) P04271 upregulates IL-1beta and P01375 expression in microglia via RAGE engagement ; and ( 3 ) P04271 /RAGE-induced upregulation of P35354 , IL-1beta and P01375 expression requires the concurrent activation of NF-kappaB and AP-1 . We also show that P04271 synergizes with IL-1beta and P01375 to upregulate on P35354 expression in microglia . Given the crucial roles of P35354 , IL-1beta and P01375 in the inflammatory response , we propose that , by engaging RAGE , P04271 might play an important role in microglia activation in the course of brain damage . Effect of p53 activity on the sensitivity of human glioblastoma cells to P09874 inhibitor in combination with topoisomerase I inhibitor or radiation . Poly ( ADP- DB01936 ) polymerase-1 ( P09874 ) is involved in the DNA repairing system by sensing and signaling the presence of DNA damage . Inhibition of P09874 is tested in combination with DNA damaging agents such as topoisomerase I inhibitors or ionizing radiations ( RT ) for the treatment of glioblastoma ( GBM ) . Disruption of p53 , widely prevalent in GBMs , plays a major role in DNA repairing system . The current study investigates whether p53 activity has an effect on the sensitivity of human GBM cells to P09874 inhibitors in combination with topoisomerase I inhibitor topotecan ( TPT ) and/or RT . Human GBM cell lines carrying a different functional status of p53 were treated with P09874 inhibitor DB02690 , in combination with TPT and/or RT . Cytotoxic effects were examined by analyzing the antiproliferative activity , the cell cycle perturbations , and the DNA damage induced by combined treatments . PARP inhibition enhanced the antiproliferative activity , the cell cycle perturbations and the DNA damage induced by both TPT or RT in GBM cells . These effects were influenced by the p53 activity : cells carrying an active p53 were more sensitive to the combination of PARP inhibitor and RT , while cells carrying an inactive p53 displayed a higher sensitivity to the combination of PARP inhibitor and TPT . Our study suggests that p53 activity influences the differential sensitivity of GBM cells to combined treatments of TPT , RT , and PARP inhibitors . © 2014 International Society for Advancement of Cytometry . Non-HLA autoimmunity genetic factors contributing to Autoimmune Polyglandular Syndrome type II in Tunisian patients . Autoimmune Polyglandular Syndrome Type II ( APSII ) is characterized by the co-occurrence of clinical insufficiency of at least two endocrine glands . Although , HLA determinants of APSII predisposition have been identified , little attention has been paid to non-HLA genes . Here , we used SNP genotyping in a Sequenom platform and genetic association tests to study a cohort of 60 APSII Tunisian patients presenting highly frequent co-occurrence of Autoimmune Thyroid Disease ( AITD ) and Type 1 Diabetes ( T1D ) and lower frequency of Addison 's disease ( AD ) . We tested the high a priori possibility that well-established non-HLA autoimmunity loci were involved in APSII and confirmed five association signals to APSII , namely : ( 1 ) two T1D-associated SNPs , in P16410 and P01589 , suggest their involvement in T1D pathogenesis in this cohort ; ( 2 ) two SNPs in Q14765 and P40933 not previously associated to endocrinopathies , are possibly involved in co-occurrence of organ autoimmunity in APSII , and ; ( 3 ) one SNP in P01375 alpha showed association to APSII irrespective of AD . While this work was performed in a relatively small cohort , these results support the notion that the non-HLA genetic component of APSII include genetic factors specific of particular autoimmune manifestations as well as genetic factors that promote the co-occurrence of multiple autoimmune endocrinopathies . 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 . Low-density lipoprotein-induced expression of interleukin-6 , a marker of human mesangial cell inflammation : effects of oxidation and modulation by lovastatin . Low-density lipoprotein ( LDL ) may contribute to the pathogenesis of glomerulosclerosis by stimulating a mesangial cell inflammatory response . P05231 ( P05231 ) is a marker of active inflammation and ongoing glomerular injury . Therefore , we investigated the effects of native and oxidized LDL on human mesangial cell production of P05231 and a possible modulation of this inflammatory response by lovastatin , which has been shown to ameliorate experimental glomerulosclerosis . Human mesangial cells were exposed for 6 or 24 h to culture medium containing either native LDL alone or a LDL mixture containing 5 or 20 % oxidized LDL . We found that native LDL stimulated 6 h mRNA expression and secretion of P05231 . This effect was further enhanced , in a dose-related manner , when mesangial cells were exposed to increasing concentrations of oxidized LDL . DB00227 markedly inhibited mesangial cell expression of P05231 mRNA and reduced P05231 secretion . The inhibitory effects of lovastatin were overridden at least partially by exogenous mevalonate . We conclude that LDL , and particularly oxidized LDL , might contribute to the pathogenesis of glomerular disease by modulating the inflammatory response of human mesangial cells , as assessed by the stimulation of P05231 expression . Moreover , this inflammatory response can be prevented by lovastatin , providing a potential direct anti-inflammatory mechanism by which P04035 inhibitors may attenuate lipid-induced glomerular injury . 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 . [ Targeted therapies in breast cancer ] . The better understanding of the biology of breast cancer has allowed the identification of new targets for anticancer therapy . DB00072 , a monoclonal antibody binding the P04626 receptor , is used since several years in the treatment of P04626 overexpressing breast cancer , including in the adjuvant setting . DB01259 , a tyrosine kinase inhibitor , was introduced more recently into the clinic . New treatment options under evaluation in P04626 overexpressing breast cancer include combinations of anti- P04626 treatments , drugs targeting the downstream signaling pathway and new anti- P04626 agents such as pertuzumab and DB05773 . This article also reviews other targeted treatments of interest in the field of breast cancer including antiangiogenic agents and drugs targeting the PI3K-AKT- P42345 pathway . Characterization of intratumoral follicular helper T cells in follicular lymphoma : role in the survival of malignant B cells . Accumulating evidences indicate that the cellular and molecular microenvironment of follicular lymphoma ( FL ) has a key role in both lymphomagenesis and patient outcome . Malignant FL B cells are found admixed to specific stromal and immune cell subsets , in particular P01730 (pos) T cells displaying phenotypic features of follicular helper T cells ( T(FH) ) . The goal of our study was to functionally characterize intratumoral P01730 (pos) T cells . We showed that P32302 (hi) Q9Y6W8 (hi) P01730 (pos) T cells sorted from FL biopsies comprise at least two separate cell populations with distinct genetic and functional features : ( i ) CD25(pos) follicular regulatory T cells ( T(FR) ) , and ( ii ) CD25(neg) T(FH) displaying a FL-B cell supportive activity without regulatory functions . Furthermore , despite their strong similarities with tonsil-derived T(FH) , purified FL-derived T(FH) displayed a specific gene expression profile including an overexpression of several genes potentially involved directly or indirectly in lymphomagenesis , in particular P01375 , P01374 , P05112 or P29965 . Interestingly , we further demonstrated that these two last signals efficiently rescued malignant B cells from spontaneous and rituximab-induced apoptosis . Altogether , our study demonstrates that tumor-infiltrating P01730 (pos) T cells are more heterogeneous than previously presumed , and underlines for the first time the crucial role of T(FH) in the complex set of cellular interactions within FL microenvironment . Radiolabeled ligand binding to the catalytic or allosteric sites of O76074 and PDE11 . Cyclic nucleotide phosphodiesterases ( PDEs ) have been investigated for years as targets for therapeutic intervention in a number of pathophysiological processes . Phosphodiesterase-5 ( O76074 ) , which is highly specific for guanosine 3'-5'-cyclic-monophosphate ( cGMP ) at both its catalytic site and its allosteric sites , has generated particular interest because it is potently and specifically inhibited by three drugs : sildenafil ( Viagra , Pfizer ) , tadalafil ( DB00820 , Lilly- Q9Y6W8 ) , and vardenafil ( DB00862 , Bayer GSK ) . Previously , we have used [(3)H]cGMP to directly study the interaction of cGMP with the allosteric sites of O76074 , but because cGMP binds with relatively low affinity to the catalytic site , it has been difficult to devise a binding assay for this particular binding reaction . This approach using measurement of radiolabeled ligand binding continues to allow us to more precisely define functional features of the enzyme . We now use a similar approach to study the characteristics of high-affinity [(3)H]inhibitor binding to the O76074 catalytic domain . For these studies , we have prepared [(3)H]sildenafil and [(3)H]tadalafil , two structurally different competitive inhibitors of O76074 . The results demonstrate that radiolabeled ligands can be used as probes for both catalytic site and allosteric site functions of O76074 . We describe herein the methods that we have established for studying the binding of radiolabeled ligands to both types of sites on O76074 . These techniques have also been successfully applied to the study of binding of radiolabeled O76074 inhibitors to PDE11 , suggesting that these methods are applicable to the study of other PDEs , and perhaps other enzyme families . An evidence based therapeutic approach to hereditary and acquired angioedema . PURPOSE OF REVIEW : Hereditary angioedema ( HAE ) due to P05155 ( DB05341 ) deficiency ( HAE- DB05341 ) , HAE with normal DB05341 , and acquired angioedema due to DB05341 deficiency are rare but important diseases that can be associated with significant morbidity and mortality . Research into the pathogenesis of angioedema has expanded greatly and has led to new clinical trials with novel therapeutic agents and strategies . RECENT FINDINGS : Strategies for managing HAE- DB05341 are aimed at treating acute attacks or preventing attacks through the use of prophylactic treatment . Agents available in Europe for treating acute attacks include plasma-derived DB05341 concentrates , a bradykinin B2 receptor ( P30411 ) antagonist , and a recombinant human DB05341 . In the USA , a plasma-derived DB05341 concentrate , a bradykinin P30411 antagonist , and a plasma kallikrein inhibitor have been approved for the treatment of acute HAE- DB05341 attacks . DB05341 concentrates and attenuated androgens are used for short-term prophylactic treatment . Long-term prophylactic treatments include attenuated androgens , a plasma-derived DB05341 concentrate , and antifibrinolytics . Plasma-derived DB05341 and a bradykinin P30411 antagonist are approved for self-administration at home . SUMMARY : The number of management options for HAE- DB05341 and similar conditions has increased considerably within the last few years , thus helping to alleviate the burden of these rare diseases . 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 . Donor pre-treatment with everolimus or cyclosporine does not reduce ischaemia-reperfusion injury in a rat kidney transplant model . BACKGROUND : Immunosuppressive agents have been investigated in renal ischaemia-reperfusion injury ( IRI ) and have frequently demonstrated a beneficial effect . Most studies focused on treatment of the recipient at the time of transplantation . Pre-treatment of these organs before injury ( pharmacological pre-conditioning ) may particularly protect these organs . This study aimed to investigate the possible protective effects of donor pre-treatment with cyclosporine ( DB00091 ) or the P42345 inhibitor everolimus or their combination against IRI during renal transplantation in a rat model . METHODS : Donors received vehicle , DB00091 ( 5 mg/kg ) , everolimus ( 0.5 mg/kg ) or CsA + everolimus . Two oral doses were administered to the donors at 24 h and again at 6 h prior to donor kidney removal . Syngeneic rat kidneys were preserved in UW solution for 24 h prior to transplantation . After 24 h of reperfusion , blood and tissue samples were collected from recipients for further analysis . RESULTS : Renal functions as determined by creatinine and necrosis scores were not different between the experimental groups . Cleaved caspase-3 , heat shock protein 70 ( HSP70 ) , tumor-necrosis factor-alpha ( P01375 -α ) and nitrotyrosine protein levels were not statistically different between the four treatment groups at 24 h post-transplantation . Blood NMR analysis on metabolic markers for IRI reveals no beneficial effects of donor pre-treatment on the 24-h outcome in transplantation . CONCLUSIONS : When given alone or as a combination to donors before organ recovery , cyclosporine or everolimus does not appear to ameliorate IRI . 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 . 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 . Variation in genes coding for AMP-activated protein kinase ( AMPK ) and breast cancer risk in the European Prospective Investigation on Cancer ( EPIC ) . AMP-activated protein kinase ( AMPK ) is an energy sensing/signalling intracellular protein which is activated by an increase in the cellular AMP: DB00171 ratio after DB00171 depletion . Once activated , AMPK inhibits fatty acid synthesis and the Akt- P42345 pathway , and activates the p53- P38936 axis . All these molecular mechanisms are thought to play a key role in breast carcinogenesis . We investigated the genetic variability of four genes encoding AMPK ( Q13131 , P54646 , Q9Y478 and O43741 ) . Using a tagging approach and selecting SNPs we covered all the common genetic variation of these genes . We tested association of tagging SNPs in our four candidate genes with breast cancer ( BC ) risk in a study of 1340 BC cases and 2536 controls nested into the European Prospective Investigation into Cancer and Nutrition ( EPIC ) . Given the relevance of AMPK on fatty acid synthesis and the importance of body fatness as a BC risk factor , we tested association of SNPs and body-mass index as well . We observed no statistically significant association between the SNPs in the PRKAs genes and BC risk and BMI after correction for multiple testing .

MH_train_1481

MH_train_1481

MH_train_1481

interacts_with DB01211?

multiple_choice

Sensitivity toward tyrosine kinase inhibitors varies between different activating mutations of the P36888 receptor . Activating mutations of P36888 have been detected in patients with acute myeloid leukemia ( AML ) . Two distinct types of P36888 mutations are most common : internal tandem duplication ( ITD ) of sequences coding for the juxtamembrane domain and point mutations at codon 835 ( Asp835 ) within the kinase domain . Both types of mutations constitutively activate the tyrosine kinase activity of P36888 in experimental systems and result in factor-independent proliferation of Ba/ P13726 and 32D cells . Recently , novel mutations within the activation loop were identified in patients with AML : deletion of isoleucine 836 ( Ile836del ) and an exchange of isoleucine 836 to methionine plus an arginine insertion ( Ile836Met+ DB00125 ) . To examine whether the Ile836 mutations result in constitutive activation of the P36888 receptor , we introduced both mutant P36888 cDNAs transiently into P29320 293 cells . Both mutant P36888 receptors were constitutively autophosphorylated in the absence of ligand and kinase activity led to constitutive activation of downstream signaling cascades as determined by activation of the P42229 ( signal transducer and activator of transcription 5 ) pathway . When stably expressed in the growth factor-dependent cell lines Ba/ P13726 and 32D , both deletion and insertion mutants led to factor-independent proliferation , indicating that both mutants have transforming capabilities . We then examined the sensitivity of the P36888 ITD , P36888 Asp835Tyr , and the novel P36888 receptor mutants toward the kinase inhibitors AG1296 , PKC412 , and SU5614 . We show that these P36888 kinase inhibitors have distinct inhibitory potencies against different activating P36888 receptor mutants . These results suggest that it may be useful to determine the exact kind of P36888 mutation when applying receptor kinase inhibitors in clinical trials . [ 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 . 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 . 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 . Eye disorders in diabetes : potential drug targets . Diabetes Mellitus ( DM ) is a serious medical problem that causes long-term systemic complications and considerable associated morbidity . DM can cause retinopathy ( O43583 ) , maculopathy , cataract , optic neuropathy , defects of eye muscles . DM is a risk factor for acute infectious conjunctivitis , bacterial keratitis , herpes virus infections and endophtalmitis . Elevated blood glucose induces structural , physiological and hormonal changes which affect retinal capillaries . O43583 is recognized by loss of pericyte function and capillary occlusions together leading to breakdown of blood-retinal barrier , edematous changes and proliferation of vessels and fibrous tissue . Depending on stage of O43583 , there are different preferable therapeutic approaches applied . In the case of ETDRS , in the area of leakage focal treatment should be performed , while panretinal photocoagulation is applied towards ischemic areas or beginning proliferations . Vitreal haemorrhage followed by fibroproliferative changes or tractional retinal detachment is treated by vitrectomy alone or in combination with ILM peeling . In pathogenesis of O43583 , P01308 Growth Factor ( DB01277 ) can play an important role in production of P15692 ( Vascular Endothelial Growth Factor ) . Hypoxia can up-regulate P15692 expression levels leading to pathologic ocular neovascularisation . An application of intravitreal corticosteroid treatment modulates vascular permeability by suppressing the production of P15692 , reducing both extracellular matrix metalloproteinase activity and basic fibroblast growth factor , decreasing major histocompatibility complex 2 Ag expression levels , and inhibiting activity of inflammatory cells . Clinical effects of treatment using intravitreal corticosteroids are evaluated by reduction of macular thickness and visual improvement . Intravitreal use of Anti- P15692 drugs , DB04895 , DB01270 and DB00112 can modify vasoproliferation , trigger macular edema , and , therefore , influence a prognosis for visual loss . Telmisartan , its potential therapeutic implications in cardiometabolic disorders . There is a growing body of evidence that the renin-angiotensin system ( DB01367 ) plays a pivotal role in the pathogenesis of cardiovascular diseases . Indeed , large clinical trials have demonstrated substantial benefit of the blockade of this system for cardiovascular-organ protection . 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 . Telmisartan will be a promising cardiometabolic sartan due to its unique peroxisome proliferator-activated receptor-gamma ( P37231 ) -inducing properties as well . In this review , we focused on telmisartan , and discussed its potential therapeutic implications in cardiometabolic disorders . Recharging oxidative protein repair : catalysis by methionine sulfoxide reductases towards their amino acid , protein , and model substrates . The sulfur-containing amino acid methionine ( DB00134 ) in its free and amino acid residue forms can be readily oxidized to the R and S diastereomers of methionine sulfoxide ( MetO ) . DB00134 sulfoxide reductases A ( Q9UJ68 ) and B ( Q9Y3D2 ) reduce MetO back to DB00134 in a stereospecific manner , acting on the S and R forms , respectively . A third Q9UBK8 type , fRMSR , reduces the R form of free MetO . Q9UJ68 and Q9Y3D2 are spread across the three domains of life , whereas fRMSR is restricted to bacteria and unicellular eukaryotes . These enzymes protect against abiotic and biotic stresses and regulate lifespan . MSRs are thiol oxidoreductases containing catalytic redox-active cysteine or selenocysteine residues , which become oxidized by the substrate , requiring regeneration for the next catalytic cycle . These enzymes can be classified according to the number of redox-active cysteines ( selenocysteines ) and the strategies to regenerate their active forms by thioredoxin and glutaredoxin systems . For each Q9UBK8 type , we review catalytic parameters for the reduction of free MetO , low molecular weight MetO-containing compounds , and oxidized proteins . Analysis of these data reinforces the concept that MSRAs reduce various types of MetO-containing substrates with similar efficiency , whereas MSRBs are specialized for the reduction of MetO in proteins . Vesiculopustular dermatosis : an uncommon side-effect of liraglutide ? DB06655 is a P43220 agonist , a novel medication for type 2 diabetes . We describe a case of pustules in a patient recently started on liraglutide . Common side effects of liraglutide are gastrointestinal disorders . Skin and tissue reactions are less well-known side effects . DB06655 could be the cause of skin eruptions in this patient , possibly by immunogenicity . 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 . Cyclical and dose-dependent responses of adult human mature oligodendrocytes to fingolimod . DB08868 is a sphingosine-1-phosphate ( Q14703 ) analogue that has been used in clinical trials as a systemic immunomodulatory therapy for multiple sclerosis . DB08868 readily accesses the central nervous system , raising the issue of its direct effects on neural cells . We assessed the effects of active fingolimod on dissociated cultures of mature , myelin-producing oligodendrocytes ( OLGs ) derived from adult human brain . Human OLGs express Q14703 receptor transcripts in relative abundance of Q9H228 > Q99500 > P21453 , with undetectable levels of O95977 . Low doses of fingolimod ( 100 pmol/L to 1 nmol/L ) induced initial membrane elaboration ( 2 days ) , subsequent retraction ( 4 days ) , and recurrence of extension with prolonged treatment ( 8 days ) . Higher doses ( 10 nmol/L to 1 mumol/L ) caused the opposite modulation of membrane dynamics . Retraction was rescued by co-treatment with the Q99500 / Q9H228 pathway antagonist , suramin , and was associated with RhoA-mediated cytoskeletal signaling . Membrane elaboration was mimicked using the P21453 agonist SEW2871 . DB08868 rescued human OLGs from serum and glucose deprivation-induced apoptosis , which was reversed with suramin co-treatment and mimicked using an Q9H228 agonist . High doses of fingolimod induced an initial down-regulation of Q9H228 mRNA levels relative to control ( 4 hours ) , subsequent up-regulation ( 2 days ) , and recurrent down-regulation ( 8 days ) . P21453 mRNA levels were inversely regulated compared with Q9H228 . These results indicate that fingolimod modulates maturity- and species-specific OLG membrane dynamics and survival responses that are directly relevant for myelin integrity . 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 . Quinolinol and peptide inhibitors of zinc protease in botulinum neurotoxin A : effects of zinc ion and peptides on inhibition . Quinolinol derivatives were found to be effective inhibitors of botulinum neurotoxin serotype A ( DB00083 ) . Studies of the inhibition and binding of 7-(phenyl(8-quinolinylamino)methyl)-8-quinolinol ( QAQ ) to the light chain domain ( DB00083 LC ) showed that QAQ is a non-competitive inhibitor for the zinc protease activity . Binding and molecular modeling studies reveal that QAQ binds to a hydrophobic pocket near the active site . Its inhibitor effect does not involve the removal of zinc ion from the light chain . A 24-mer P60880 peptide containing E183 to G206 with Q197C mutation ( Peptide C ) binds to DB00083 LC with an unusually slow second order binding rate constant of 76.7M(-1)s(-1) . QAQ binds to Zn(2+)-free DB00083 LC with a K(D) of 0.67microM and to Peptide C- DB00083 LC complex with a K(D) of 2.33microM . The insights of the interactions of quinolinols and peptides with the zinc protease of DB00083 should aid in the development of inhibitors of metalloproteases . Molecular mechanisms of cell proliferation induced by low power laser irradiation . Low power laser irradiation ( LPLI ) promotes proliferation of multiple cells , which ( especially red and near infrared light ) is mainly through the activation of mitochondrial respiratory chain and the initiation of cellular signaling . Recently , the signaling proteins involved in LPLI-induced proliferation merit special attention , some of which are regulated by mitochondrial signaling . P08581 ( c- DB00134 ) , a member of tyrosine protein kinase receptors ( TPKR ) , is phosphorylated during LPLI-induced proliferation , but tumor necrosis factor alpha ( P01375 ) receptor has not been affected . Activated TPKR could activate its downstream signaling elements , like Ras/Raf/MEK/ P29323 , PI3K/Akt/ P06730 , PI3K/Akt/ P29474 and P98160 -gamma/PKC pathways . Other two pathways , DeltaPsim/ DB00171 / DB02527 /JNK/AP-1 and ROS/Src , are also involved in LPLI-induced proliferation . LPLI-induced cell cycle progression can be regulated by the activation or elevated expressions of cell cycle-specific proteins . Furthermore , LPLI induces the synthesis or release of many molecules , like growth factors , interleukins , inflammatory cytokines and others , which are related to promotive effects of LPLI . Physical link and functional coupling of presynaptic calcium channels and the synaptic vesicle docking/fusion machinery . N- and P/Q-type calcium channels are localized in high density in presynaptic nerve terminals and are crucial elements in neuronal excitation-secretion coupling . In addition to mediating Ca2+ entry to initiate transmitter release , they are thought to interact directly with proteins of the synaptic vesicle docking/fusion machinery . As outlined in the preceding article , these calcium channels can be purified from brain as a complex with SNARE proteins which are involved in exocytosis . In addition , N-type and P/Q-type calcium channels are co-localized with syntaxin in high-density clusters in nerve terminals . Here we review the role of the synaptic protein interaction ( synprint ) sites in the intracellular loop II-III ( L(II-III) ) of both alpha1B and alpha1A subunits of N-type and P/Q-type calcium channels , which bind to syntaxin , P60880 , and synaptotagmin . DB01373 has a biphasic effect on the interactions of N-type calcium channels with SNARE complexes , stimulating optimal binding in the range of 10-20 microM . PKC or P62158 KII phosphorylation of the N-type synprint peptide inhibits interactions with native brain SNARE complexes containing syntaxin and P60880 . Introduction of the synprint peptides into presynaptic superior cervical ganglion neurons reversibly inhibits EPSPs from synchronous transmitter release by 42 % . At physiological Ca2+ concentrations , synprint peptides cause an approximate 25 % reduction in transmitter release of injected frog neuromuscular junction in cultures , consistent with detachment of 70 % of the docked vesicles from calcium channels based on a theoretical model . Together , these studies suggest that presynaptic calcium channels not only provide the calcium signal required by the exocytotic machinery , but also contain structural elements that are integral to vesicle docking , priming , and fusion processes . 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 . Stimulation of cloned human glucagon-like peptide 1 receptor expressed in P29320 293 cells induces DB02527 -dependent activation of calcium-induced calcium release . The actions of glucagon-like peptide-1(7-36)amide ( P0C6A0 (7-36)amide ) on cellular signalling were studied in human embryonal kidney 293 ( P29320 293 ) cells stably transfected with the cloned human P43220 . The cloned P43220 showed a single high-affinity binding site ( Kd = 0.76 nM ) . Binding of P0C6A0 (7-36)amide stimulated DB02527 production in a dose-dependent manner ( EC50 = 0.015 nM ) and caused an increase in the intracellular free Ca2+ concentration ( [Ca2+]i ) . The latter effect reflected Ca(2+)-induced Ca2+ release and was suppressed by ryanodine . We propose that the ability of P0C6A0 (7-36)amide to increase [Ca2+]i results from sensitization of the ryanodine receptors by a protein kinase A dependent mechanism . Hybrid molecules inhibiting myeloperoxidase activity and serotonin reuptake : a possible new approach of major depressive disorders with inflammatory syndrome . OBJECTIVES : Major depressive disorder ( MDD ) is accompanied with an imbalance in the immune system and cardiovascular impairments , such as atherosclerosis . Several mechanisms have been pointed out to underlie this rather unexpected association , and among them the activity of myeloperoxidase ( P05164 ) . The aim of our study was to find compounds that inhibit both P05164 and serotonin transporter ( P31645 ) for treating MDD associated with cardiovascular diseases . METHODS : P31645 inhibition was assessed with measuring of [ ( 3 ) H ] -serotonin uptake using P29320 -293 Q9UBK8 cells . P05164 inhibition was determined by taurine chloramine test on 3-(aminoalkyl)-5-fluoroindole derivatives and on clinically relevant antidepressants . All kinetic measurements were performed using a temperature-controlled stopped-flow apparatus ( model SX-18 MV ) . Promising lead compounds were docked onto P31645 3D structure modelled using the LeuT structure complexed to tryptophan ( PDB code 3F3A ) . Their toxicological profile was also assessed . KEY FINDINGS : 3-(aminoalkyl)-5-fluoroindole derivative with 5 carbons on the side chain and paroxetine showed the best activity on both P05164 and P31645 at the nanomolar range . Paroxetine was found to be the first irreversible P05164 inhibitor at nanomolar concentrations . CONCLUSIONS : Our results put forward the first hybrid molecule ( compound 25 ) and drug ( paroxetine ) that can be especially used in MDD associated with inflammatory syndrome . 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 . Peroxisome-proliferator activator receptor-gamma activation decreases attachment of endometrial cells to peritoneal mesothelial cells in an in vitro model of the early endometriotic lesion . The aim of this study was to investigate whether peroxisome proliferator-activated receptor ( Q07869 ) -gamma activation has an effect on the attachment of endometrial cells to peritoneal mesothelial cells in a well-established in vitro model of the early endometriotic lesion . The endometrial epithelial cell line EM42 and mesothelial cell line LP9 were used for this study . EM42 cells , LP9 cells or both were treated with the P37231 agonist ciglitazone ( CTZ ) at varying concentrations ( 10 , 20 and 40 microM ) x 48 h with subsequent co-culture of EM42 and LP9 cells . The rate of EM42 attachment and invasion through LP9 cells was then assessed and compared with control ( EM42 and LP9 cells co-cultured without prior treatment with CTZ ) . Next , attachment of CTZ-treated and untreated EM42 cells to hyaluronic acid ( HA ) , a cell adhesion molecule ( P62158 ) on peritoneal mesothelial cells , were assessed . Although there was no difference in EM42 attachment when LP9 cells alone were treated with CTZ , treatment of EM42 cells with 40 microM CTZ decreased EM42 attachment to LP9 cells by 27 % ( P < 0.01 ) . Treatment of both EM42 and LP9 cells with 40 microM CTZ decreased EM42 attachment to LP9 by 37 % ( P < 0.01 ) . Treatment of EM42 cells with 40 microM CTZ decreased attachment to HA by 66 % ( P = 0.056 ) . CTZ did not decrease invasion of EM42 cells through the LP9 monolayer . CTZ may inhibit EM42 cell proliferation . In conclusion , CTZ significantly decreased EM42 attachment to LP9 cells and HA in an in vitro model of the early endometriotic lesion . Evaluation of commercial antibodies against human sphingosine-1-phosphate receptor 1 . DB03203 -1-phosphate receptor 1 ( P21453 ) , also called endothelial differentiation gene 1 , plays an important role in migration , proliferation , and survival of several types of cells including endothelial cells and lymphocytes and is involved in multiple sclerosis . Two commercial rabbit anti- P21453 antibodies ( polyclonal and monoclonal ) were tested on CHO cells expressing P21453 receptors fused to the green fluorescent protein at the C-terminal end and on Pichia pastoris and P29320 cells expressing cmyc-tagged P21453 . Polyclonal antibodies did not give any signal by Western blot , immunofluorescence , and flow cytofluorometry . Monoclonal antibodies were able to reveal an unspecific band by Western blot performed on various cell types . Consequently , in our hands and using our protocols , we show that these antibodies did not specifically detect P21453 receptors . DB05213 is a uniquely potent and selective inhibitor of P36888 for the treatment of acute myeloid leukemia ( AML ) . Activating mutations in the receptor tyrosine kinase P36888 are present in up to approximately 30 % of acute myeloid leukemia ( AML ) patients , implicating P36888 as a driver of the disease and therefore as a target for therapy . We report the characterization of DB05213 , a second-generation P36888 inhibitor , and a comparison of DB05213 with the first-generation P36888 inhibitors CEP-701 , DB05465 , PKC-412 , sorafenib , and sunitinib . DB05213 exhibits low nanomolar potency in biochemical and cellular assays and exceptional kinase selectivity , and in animal models is efficacious at doses as low as 1 mg/kg given orally once daily . The data reveal that the combination of excellent potency , selectivity , and pharmacokinetic properties is unique to DB05213 , which therefore is the first drug candidate with a profile that matches the characteristics desirable for a clinical P36888 inhibitor .

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SC-435 , an ileal apical sodium-codependent bile acid transporter inhibitor alters mRNA levels and enzyme activities of selected genes involved in hepatic cholesterol and lipoprotein metabolism in guinea pigs . We have demonstrated that SC-435 , an apical sodium codependent bile acid transporter ( Q12908 ) inhibitor , lowers plasma low-density lipoprotein cholesterol ( LDL-C ) concentrations in guinea pigs . The purpose of this study was to further examine the hypocholesterolemic effects of SC-435 , by measuring the activity and RNA expression of regulatory enzymes of hepatic cholesterol and lipoprotein metabolism . In addition , the use of a combination ( COMBO ) therapy with simvastatin , a 3-hydroxy-3-methylglutaryl-coenzyme A ( HMG- DB01992 ) reductase inhibitor , was also tested . Male Hartley guinea pigs were randomly allocated to one of three diets ( n=10 per group ) , for 12 weeks . The control diet contained no Q12908 inhibitor or simvastatin . The monotherapy diet ( ASBTi ) contained 0.1 % of SC-435 . The COMBO therapy consisted of a lower dose of SC-435 ( 0.03 % ) and 0.05 % simvastatin . DB04540 ester transfer protein ( P11597 ) and P04035 mRNA abundance were determined using RT-PCR techniques . Hepatic P04035 and cholesterol 7alpha-hydroxylase ( P22680 ) activities were measured by radioisotopic methods . Compared to the control group , P11597 activity was 34 % and 56 % lower with ASBTi and COMBO , respectively . Similarly , P11597 mRNA expression was reduced by 36 % and 73 % in ASBTi and COMBO groups , respectively . DB04540 7alpha-hydroxylase and P04035 activities were increased approximately 2-fold with ASBTi and COMBO treatments , respectively . Likewise , P04035 mRNA expression was increased 33 % with ASBTi treatment . These results suggest that both SC-435 monotherapy and combination therapy lower LDL cholesterol concentrations by altering both hepatic cholesterol homeostasis and the intravascular processing of lipoproteins in guinea pigs . Huge clusters of embryonic stem cells in human embryos : a morphologic study . BACKGROUND : Nothing is known about huge clusters ( HC ) of embryonic stem cells ( ESC ) in human fetal organs ( HFO ) . AIM : To know the status of HC-ESC in HFO . METHODS : Morphology and immunohistochemistry ( IHC ) in 32 HFO of 7-40 gestational weeks ( GW ) . RESULTS : HC-ESC were seen in many HFO including central nervous system , spinal cords , spine , soft tissue , bone , skin , thyroid , lung , liver , pancreas , gall bladder , extrahepatic bile duct , adrenal , kidney , bladder , foregut , midgut , hindgut , female and male genital organs , and neurons . HC-ESC 's were composed of two populations depending on constituting cells . One were large cells with ample acidophilic cytoplasms with vesicular nuclei and nucleoli . The other were small cells with scant cytoplasm with hyperchromatic nuclei without nucleoli , resembling lymphocytes . The HC-ESC were frequently showed neuronal differentiation . HC-ESC were positive for P13591 , synaptophysin , P09104 , chromogranin , P16234 , AFP , ErbB2 , bcl-2 , P10721 , MET . They were negative for P08575 , CD3 , P11836 , P15941 , P06731 , CA19-9 , cytokeratin ( CK ) 7 , CK8 , CK18 , CK19 , P15941 , Q02817 , P98088 , and Q6W4X9 . The mean Ki-67 labeling index ( LI ) was 13 % ± 7 % . HC-ESC showed a little glycogen but lacked mucins . These HC-ESC were seen in 7-25 GW , and they were rarely seen in 26-40 GW . CONCLUSIONS : The morphology , IHC , and ontogeny of HC-ESC were described . The inhibitory effect of the leukotriene receptor antagonist on leukotriene D4-induced Q02817 /5AC gene expression and mucin secretion in human airway epithelial cells . OBJECTIVES : Mucin gene expression and mucin production are markedly increased in inflammatory airway disorders such as asthma , chronic bronchitis and rhinosinusitis . Cytokines , lipopolysaccharides and other inflammatory mediators such as prostaglandin and leukotriene are related to the secretion and production of mucin . However , the relationship of leukotrienes with mucin genes expression is not clear . The aim of this study is to evaluate Q02817 /5AC gene expression and mucin secretion by the leukotriene receptor in human airway epithelial cells . METHODS : The effect of leukotriene D(4) and the leukotriene receptor antagonist , pranlukast hydrate ( ONO-1078 ) on the regulation of Q02817 /5AC gene expression and mucin secretion were observed in human airway NCI-H292 epithelial cells . The mRNA levels of Q02817 /5AC and the amount of mucin were determined by reverse transcription-polymerase chain reaction ( RT-PCR ) and immunoassay . RESULTS : Leukotriene D(4) upregulated Q02817 /5AC gene expression and mucin secretion in a dose dependent pattern . DB01411 hydrate ( ONO-1078 , 100 microM ) downregulated the leukotriene D(4)-induced Q02817 /5AC gene expression and mucin secretion . CONCLUSION : These results suggest that the leukotriene receptor system is one of the mechanisms related to Q02817 /5AC gene expression and mucin secretion in the human airway epithelium . A critical importance of polyamine site in DB01221 receptors for neurite outgrowth and fasciculation at early stages of P19 neuronal differentiation . We have investigated the role of N-methyl-d-aspartate receptors ( NMDARs ) and gamma-aminobutyric acid receptors type A ( GABA(A)Rs ) at an early stage of P19 neuronal differentiation . The subunit expression was profiled in 24-hour intervals with RT-PCR and functionality of the receptors was verified via fluo-3 imaging of Ca(2+) dynamics in the immature P19 neurons showing that both DB01221 and GABA excite neuronal bodies , but only polyamine-site sensitive NMDAR stimulation leads to enhanced Ca(2+) signaling in the growth cones . Inhibition of Q9UHB4 / Q13224 NMDARs by 1 muM ifenprodil severely impaired P19 neurite extension and fasciculation , and this negative effect was fully reversible by polyamine addition . In contrast , GABA(A)R antagonism by a high dose of 200 microM bicuculline had no observable effect on P19 neuronal differentiation and fasciculation . Except for the differential NMDAR and GABA(A)R profiles of Ca(2+) signaling within the immature P19 neurons , we have also shown that inhibition of Q9UHB4 / Q13224 NMDARs strongly decreased mRNA level of P13591 -180 , which has been previously implicated as a regulator of neuronal growth cone protrusion and neurite extension . Our data thus suggest a critical role of Q9UHB4 / Q13224 NMDARs during the process of neuritogenesis and fasciculation of P19 neurons via differential control of local growth cone Ca(2+) surges and P13591 -180 signaling . Radiation hybrid map of 13 loci on the long arm of chromosome 5 . Radiation hybrid mapping was used in conjunction with a natural deletion mapping panel to predict the order of and distance between 13 loci in the distal portion of the long arm of human chromosome 5 . A panel of irradiation hybrids containing fragments of 5q was generated from an P00492 + Chinese hamster-human cell hybrid containing a derivative chromosome 5 [ der(5)t(4;5) ( 5qter ---- 5p15.1::4p15.1 ---- 4pter ) ] as its only human DNA . One hundred nine radiation hybrids containing human DNA were screened with polymerase chain reaction primer sets representing nine genes encoding growth factors , growth factor receptors , or hormone receptors ( P08700 , P05112 , P05113 , P07333 , P05230 , P07550 , GRL , P14867 , and P21728 ) as well as four other loci ( P16591 , P09486 , P62263 , and P08571 ) to generate a radiation hybrid map of the area 5q21-q35 . A physical map predicting the order of and distance between the 13 loci was constructed based on segregation of the 13 loci in hybrid clones . The radiation hybrid panel will be useful as a mapping tool for determining the location and order of other genes and polymorphic loci in this region as well as for generating new DNA probes from specific regions . Ubiquitination regulates the assembly of VLDL in HepG2 cells and is the committing step of the apoB-100 ERAD pathway . P04114 ( apoB-100 ) is degraded by endoplasmic reticulum-associated degradation ( ERAD ) when lipid availability limits assembly of VLDLs . The ubiquitin ligase Q9UKV5 and the AAA-ATPase p97 have been implicated in the proteasomal degradation of apoB-100 . To study the relationship between ERAD and VLDL assembly , we used small interfering RNA ( siRNA ) to reduce Q9UKV5 expression in HepG2 cells . Reduction of Q9UKV5 decreased apoB-100 ubiquitination and cytosolic apoB-ubiquitin conjugates . Radiolabeling studies revealed that Q9UKV5 knockdown increased secretion of newly synthesized apoB-100 and , unexpectedly , enhanced VLDL assembly , as the shift in apoB-100 density in Q9UKV5 -reduced cells was accompanied by increased triacylglycerol ( TG ) secretion . To explore the mechanisms by which Q9UKV5 reduction might enhance VLDL assembly , we compared the effects of Q9UKV5 knockdown with those of U0126 , a mitogen-activated protein kinase/ P29323 kinase1/2 inhibitor that enhances apoB-100 secretion in HepG2 cells . U0126 treatment increased secretion of both apoB100 and TG and decreased the ubiquitination and cellular accumu-lation of apoB-100 . Furthermore , p97 knockdown caused apoB-100 to accumulate in the cell , but if Q9UKV5 was concomitantly reduced or assembly was enhanced by U0126 treatment , cellular apoB-100 returned toward baseline . This indicates that ubiquitination commits apoB-100 to p97-mediated retrotranslocation during ERAD . Thus , decreasing ubiquitination of apoB-100 enhances VLDL assembly , whereas improving apoB-100 lipidation decreases its ubiquitination , suggesting that ubiquitination has a regulatory role in VLDL assembly . 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 . Differential regulation of P04035 and Insig-1 by enzymes of the ubiquitin-proteasome system . The endoplasmic reticulum ( ER ) -resident enzyme 3-hydroxy-3-methylglutaryl DB01992 ( HMG- DB01992 ) reductase catalyzes the rate-limiting step in sterol production and is the therapeutic target of statins . Understanding P04035 regulation has tremendous implications for atherosclerosis . P04035 levels are regulated in response to sterols both transcriptionally , through a complex regulatory loop involving the ER Insig proteins , and posttranslationally , by Insig-dependent protein degradation by the ubiquitin-proteasome system . The ubiquitin ligase ( E3 ) Q9UKV5 has been implicated in the sterol-regulated degradation of P04035 and Insig-1 through ER-associated degradation ( ERAD ) . More recently , a second ERAD E3 , Q8WU17 , has also been reported to play a role in the sterol-accelerated degradation of P04035 . We interrogated this network in Q9UKV5 (-/-) mouse embryonic fibroblasts and also assessed two fibroblast cell lines using RNA interference . Although we consistently observe involvement of Q9UKV5 in Insig-1 degradation , we find no substantive evidence to support roles for either Q9UKV5 or Q8WU17 in the robust sterol-accelerated degradation of P04035 . We discuss factors that might lead to such discrepant findings . Our results suggest a need for additional studies before definitive mechanistic conclusions are drawn that might set the stage for development of drugs to manipulate Q9UKV5 function in metabolic disorders . DB01373 -binding protein of chorioallantoic membrane : identification and development expression . A calcium-binding protein ( CaBP ) which has a molecular weight of 100,000 and an isoelectric point of 8.0 has been isolated from the chorioallantoic membrane ( P62158 ) of the chick embryo . Expression of the CaBP occurs simultaneously with the onset of calcium absorption from the egg shell by the P62158 , and the temporal increase in CaBP activity is coincident with calcium deposition in the embryo . The P62158 CaBP differs in properties from the CaBPs of the adult organs , including the vitamin-D-dependent protein of intestine . Differential effects of amfonelic acid on the haloperidol- and clozapine-induced increase in extracellular DOPAC in the nucleus accumbens and the striatum . This study compares the effects of the nonamphetamine stimulant amfonelic acid on the increase in extracellular 3,4-dihydroxyphenylacetic acid ( DOPAC ) induced by haloperidol and clozapine in the nucleus accumbens and the striatum of anaesthetized rats . DOPAC was simultaneously recorded in both regions using differential pulse voltammetry with electrically pretreated carbon fibre electrodes . Amfonelic acid ( 2.5 mg/kg s.c. ) did not alter basal striatal DOPAC but produced a significant reduction in extracellular DOPAC in the nucleus accumbens . DB00502 ( 1 mg/kg s.c. ) increased extracellular DOPAC in both regions . When amfonelic acid was injected 5 min before haloperidol , the increase in DOPAC was potentiated in both the nucleus accumbens and the striatum but with a greater effect in the striatum . Clozapine ( 30 mg/kg i.p. ) increased extracellular DOPAC in both regions , an effect partially attenuated by amfonelic acid in both regions but to a greater extent in the striatum . When ritanserin ( 5 mg/kg i.p. ) , a serotonergic antagonist ( P28223 ) , was co-administered with haloperidol , the potentiation by amfonelic acid of the increase in extracellular DOPAC induced by haloperidol was attenuated in both the nucleus accumbens and the striatum . The present results confirm that amfonelic acid can be used to discriminate neurochemically between haloperidol and clozapine in vivo . The effects of amfonelic acid on the neuroleptic-induced changes in extracellular DOPAC were greater in the striatum than the nucleus accumbens . These results further demonstrate that both neuroleptics increase dopamine metabolism in the two brain regions but by different mechanisms , supporting the view that the regulation of dopamine metabolism differs in the two regions. ( ABSTRACT TRUNCATED AT 250 WORDS ) Molecular basis of differential resistance to cycloguanil and pyrimethamine in Plasmodium falciparum malaria . DB01131 and pyrimethamine are antifolate drugs with distinct chemical structures that are used commonly in the prophylaxis and treatment of Plasmodium falciparum malaria . Clinical reports and field studies have suggested that some parasites refractory to proguanil can be treated with pyrimethamine , and vice versa . Analysis of the P. falciparum dihydrofolate reductase ( P00374 ) from different parasites reveals the structural basis for differential susceptibility to these antifolate drugs . Parasites harboring a pair of point mutations from Ala-16 to DB00161 -16 and from DB00133 -108 to DB00156 -108 are resistant to cycloguanil ( the active metabolite of proguanil ) but not to pyrimethamine . A single DB00174 -108 mutation , on the other hand , confers resistance to pyrimethamine with only a moderate decrease in susceptibility to cycloguanil . Significant cross-resistance to both drugs occurs in parasites having mutations that include DB00133 -108 ---- DB00174 -108 and DB00167 -164 ---- DB00149 -164 . These results reflect the distinct structures of pyrimethamine and cycloguanil and suggest fine differences in binding within the active site cavity of P00374 . Alternative inhibitors , used alone or in combination , may be effective against some strains of cycloguanil- or pyrimethamine-resistant malaria . 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 . Reviews of anti-infective agents : maraviroc : the first of a new class of antiretroviral agents . DB04835 is the first US Food and Drug Administration-approved drug from a new class of antiretroviral agents that targets a host protein , the chemokine receptor P51681 , rather than a viral target . Binding of maraviroc to this cell-surface protein results in blocking human immunodeficiency virus type 1 ( HIV-1 ) attachment to the coreceptor and prevents the virus from entering P01730 + cells . In this review , we include the details of the discoveries that led to the development of this drug . The drug 's pharmacology , including pharmacokinetics and drug interactions , is discussed , as are the clinical efficacy studies that led to licensure . HIV-1 mechanisms of resistance to maraviroc , assays to determine viral coreceptor use ( tropism ) , drug safety , and clinical use of maraviroc are discussed at length . DB06698 treatment in managing vertigo and improving vestibular compensation : clarification . DB06698 dihydrochloride ( betahistine ) is currently used in the management of vertigo and vestibular pathologies with different aetiologies . The main goal of this review is to clarify the mechanisms of action of this drug , responsible for the symptomatic relief of vertigo and the improvement of vestibular compensation . The review starts with a brief summary recalling the role of histamine as a neuromodulator/neurotransmitter in the control of the vestibular functions , and the role of the histaminergic system in vestibular compensation . Then are presented data recorded in animal models demonstrating that betahistine efficacy can be explained by mechanisms targeting the histamine receptors ( HRs ) at three different levels : the vascular tree , with an increase of cochlear and vestibular blood flow involving the P35367 ; the central nervous system , with an increase of histamine turnover implicating the Q9Y5N1 , and the peripheral labyrinth , with a decrease of vestibular input implying the Q9Y5N1 / Q9H3N8 . Clinical data from vestibular loss patients show the impact of betahistine treatment for the long-term control of vertigo , improvement of balance and quality of life that can be explained by these mechanisms of action . However , two conditions , at least , are required for reaching the betahistine therapeutic effect : the dose and the duration of treatment . Experimental and clinical data supporting these requirements are exposed in the last part of this review . [ 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 . 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 . 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 . Cdk5 inhibitor roscovitine alleviates neuropathic pain in the dorsal root ganglia by downregulating N-methyl-D-aspartate receptor subunit 2A . P12004 -dependent kinase 5 ( Cdk5 ) is a member of the small proline-directed serine/threonine kinase family . Cdk5 is not involved in cell cycle regulation , but is implicated in neurodegenerative disorders . However , the role of Cdk5 in neuropathic pain remains unclear . This study aimed to evaluate the possibility that Cdk5 is involved in neuropathic pain in the dorsal root ganglia ( Q86YR7 ) . We injected intrathecally Cdk5 inhibitor roscovitine in rat model of chronic compression of dorsal root ganglion and examined pain behaviors and the expression of N-methyl-d-aspartate receptor subunit 2A ( Q12879 ) but not Q13224 or Q9UHB4 in Q86YR7 . We found that roscovitine alleviated neuropathic pain , causing decline in paw withdrawal mechanical threshold and paw withdrawal thermal latency . Furthermore , roscovitine inhibited Q12879 expression in Q86YR7 . These data suggest that Cdk5- Q12879 pathway regulates neuropathic pain in Q86YR7 , and intrathecal injection of roscovitine could alleviate neuropathic pain . Our findings provide new insight into the analgesic effects of Roscovitine and identify Cdk5- Q12879 pathway as a potential target for effective treatment of neuropathic pain . Prediction of P24385 amplification using plasma DNA as a prognostic marker in oesophageal squamous cell carcinoma . BACKGROUND : We aimed to develop a new biomarker to predict cyclin D1 ( P24385 ) status using plasma DNA in oesophageal squamous cell carcinoma ( ESCC ) patients . METHODS : We evaluated the ratio of the P24385 ( 11q13 ) dosage to the dopamine receptor D2 ( P14416 ; 11q22-23 ) dosage ( C/D ratio ) as P24385 copy number . This study was divided into three steps : ( 1 ) Determination of a cutoff value for the C/D ratio in test scale ; ( 2 ) Comparison of the C/D ratio in between plasma samples and cancer tissues in ESCC patients showing high plasma C/D ratio ; ( 3 ) Validation study of the clinical application of the plasma C/D ratio as a diagnostic and prognostic marker , by comparing with clinicopathologic factors in 96 ESCC patients . RESULTS : The plasma C/D ratio was significantly higher in the ESCC group than the controls ( P=0.0134 ) . A high plasma C/D ratio reflected the tumour C/D ratio , and significantly correlated with a poorer prognosis ( P=0.0186 ) . Moreover , the high C/D ratio was found to be an independent prognostic factor on multivariate analysis ( P=0.0266 ; hazard ratio 5.988 ) . CONCLUSION : Prediction of P24385 amplification using plasma DNA is thought to be a promising prognostic biomarker in ESCC patients . Novel structural features of CDK inhibition revealed by an ab initio computational method combined with dynamic simulations . The rational development of specific inhibitors for the approximately 500 protein kinases encoded in the human genome is impeded by a poor understanding of the structural basis for the activity and selectivity of small molecules that compete for DB00171 binding . Combining classical dynamic simulations with a novel ab initio computational approach linear-scalable to molecular interactions involving thousands of atoms , we have investigated the binding of five distinct inhibitors to the cyclin-dependent kinase P24941 . We report here that polarization and dynamic hydrogen bonding effects , so far undetected by crystallography , affect both their activity and selectivity . The effects arise from the specific solvation patterns of water molecules in the DB00171 binding pocket or the intermittent formation of hydrogen bonds during the dynamics of CDK/inhibitor interactions and explain the unexpectedly high potency of certain inhibitors such as 3-(3H-imidazol-4-ylmethylene)-5-methoxy-1,3-dihydro-indol-2-one ( DB03428 ) . The Lys89 residue in the DB00171 -binding pocket of P24941 is observed to form temporary hydrogen bonds with the three most potent inhibitors . This residue is replaced in P11802 by Thr89 , whose shorter side-chain can not form similar bonds , explaining the relative selectivity of the inhibitors for P24941 . Our results provide a generally applicable computational method for the analysis of biomolecular structures and reveal hitherto unrecognized features of the interaction between protein kinases and their inhibitors . 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 . Antitumor activity of arsenic trioxide on retinoblastoma : cell differentiation and apoptosis depending on arsenic trioxide concentration . PURPOSE : DB01169 ( ATO ) targets multiple pathways in malignant cells , resulting in the promotion of differentiation or in the induction of apoptosis . The antitumor activity of ATO on retinoblastoma was investigated . METHODS : Human retinoblastoma cells were incubated with various ATO concentrations . The antiproliferative effect of ATO was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay , and the effect of ATO on cell-cycle progression was validated by flow cytometry . At a low concentration , the ATO-induced differentiation of retinoblastoma cells was evaluated by neurofilament expression and extracellular signal-regulated kinase ( P29323 )1/2 activation , which was confirmed by the inhibition of P27361 /2 . At a high concentration , ATO-induced H(2)O(2) production was investigated with the cell-permeable fluorescent dye 2'7'-dichlorofluorescein-diacetate , and the relationship of ATO-induced H(2)O(2) production with caspase-3-dependent apoptosis was validated by Western blot and 4'6-diamidino-2-phenolindole staining , which was confirmed by reactive oxygen species ( ROS ) inhibition . The effect of ATO on tumor formation was assessed with an orthotopic animal model of retinoblastoma . RESULTS : The antitumor activity of ATO in retinoblastoma was related to two main mechanisms , differentiation and apoptosis , which were determined by the level of ATO . At a low dose ( < or= 1 microM ) , ATO induced the differentiation of retinoblastoma cells through P27361 /2 activation , whereas ROS generation by a high dose ( > or= 2 microM ) of ATO induced apoptosis in retinoblastoma cells . Moreover , ATO at low and high doses effectively inhibited tumor formation . CONCLUSIONS : These results suggest that ATO can be used as an effective alternative therapeutic for the treatment of retinoblastoma . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . P35372 -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 . Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptides ( PACAP27 ) and PACAP38 ) protect P01730 +CD8+ thymocytes from glucocorticoid-induced apoptosis . In the present study , the effects of vasoactive intestinal peptide ( P01282 ) and the pituitary adenylate cyclase-activating polypeptides , PACAP27 and PACAP38 , in a concentration range from 10(-13) to 10(-6) mol/L were studied in vitro on the spontaneous and dexamethasone ( DEX ) -induced apoptosis in rat thymocytes . The results show that P01282 and both PACAPs inhibit significantly and in a similar way the DNA fragmentation characteristic of glucocorticoid-induced apoptosis and increase the cell survival of thymocytes , with a maximal effect observed at 10(-8) to 10(-9) mol/L . This study showed the ability of the P01282 -receptor ( P01282 -R ) antagonist [ N-Ac-Tyr1,D-Phe2 ] - P01286 (1-29) amide to partially reverse the inhibitory effect of P01282 and both PACAPs on DEX-induced apoptosis , providing evidence for a specific Q6PFW1 -R-mediated response and supporting the involvement of a single receptor for the three neuropeptides . Phenotypic analysis showed that P01282 , PACAP27 , and PACAP38 protect predominantly P01730 +CD8+ thymocytes from glucocorticoid-induced apoptosis . These findings suggest that these neuropeptides could be involved in intrathymic T-cell maturation . 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 . 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 . DB01131 resistance in Plasmodium falciparum African isolates : assessment by mutation-specific polymerase chain reaction and in vitro susceptibility testing . The antifolate proguanil is commonly used in the prophylaxis and treatment of Plasmodium falciparum malaria . A series of point mutations in the dihydrofolate reductase ( P00374 ) gene has been linked to differential susceptibility of varied P. falciparum clones or isolates to this drug . To survey the efficiency of proguanil prophylaxis in an African endemic region , and to evaluate the level of proguanil resistance in the corresponding parasite population , we performed drug susceptibility assays with P. falciparum isolates from Senegal , Kenya , and Niger . In parallel , we developed a mutation-specific polymerase chain reaction assay that enabled us to characterize mutations in the P00374 gene of the same isolates without in vitro parasite cultivation . We confirm previously available data showing that parasites harboring a point mutation from Ser108 to DB00174 present a decrease in susceptibility to cycloguanil ( the active metabolite of proguanil ) , and we show that mutations in codons 51 and 59 appear to modulate the level of resistance to cycloguanil . No mutations in codons 16 and 164 were detected in resistant parasites , in contrast with results from some previous studies . Dynorphin up-regulation in the dentate granule cell mossy fiber pathway following chronic inhibition of Q13224 -containing NMDAR is associated with increased CREB ( DB00133 133 ) phosphorylation , but is independent of P23560 /TrkB signaling pathways . Emerging evidence suggests that neuronal responses to N-methyl-d-aspartate ( NMDAR ) activation/inactivation are influenced by subunit composition . For example , activation of synaptic NMDAR ( comprised of Q12879 > Q13224 ) phosphorylates DB02527 -response-element-binding protein ( CREB ) at DB00133 133 , induces P23560 expression and promotes neuronal survival . Activation of extrasynaptic NMDAR ( comprised of Q13224 > GluN2 ) dephosphorylates CREB ( DB00133 133 ) , reduces P23560 expression and triggers neuronal death . These results led us to hypothesize that chronic inhibition of Q13224 -containing NMDAR would increase CREB ( DB00133 133 ) phosphorylation , increase P23560 levels and subsequently alter downstream dynorphin ( DYN ) and neuropeptide Y ( P01303 ) expression . We focused on DYN and P01303 because these neuropeptides can decrease excitatory neurotransmission and seizure occurrence and we reported previously that seizure-like events are reduced following chronic treatment with Q13224 antagonists . Consistent with our hypothesis , chronic treatment ( 17-21days ) of hippocampal slice cultures with the Q13224 -selective antagonists ifenprodil or Ro25,6981 increased both CREB ( DB00133 133 ) phosphorylation and granule cell mossy fiber pathway DYN expression . Similar treatment with the non-subtype-selective NMDAR antagonists d-APV or memantine had no significant effect on either CREB ( DB00133 133 ) phosphorylation or DYN expression . In contrast to our hypothesis , P23560 levels were decreased following chronic treatment with Ro25,6981 , but not ifenprodil , d-APV or memantine . Blockade of P23560 actions and TrkB activation did not significantly augment hilar DYN expression in vehicle-treated cultures and had no effect in Ro25,6981 treated cultures . These findings suggest that chronic exposure to Q13224 -selective NMDAR antagonists increased DYN expression through a putatively pCREB-dependent , but P23560 /TrkB-independent mechanism . DB01411 , a leukotriene receptor antagonist , inhibits interleukin-5 production via a mechanism distinct from leukotriene receptor antagonism . BACKGROUND : DB01411 , a cysteinyl leukotriene receptor 1 ( Q9Y271 ) antagonist , inhibits not only airway smooth muscle contraction , but also allergic inflammation . The aim of this study was to determine the mechanism of pranlukast-induced interleukin-5 ( P05113 ) inhibition in allergic inflammation . METHODS : Surgically resected human lung tissue was passively sensitized in vitro with mite-allergen-sensitized sera , followed by stimulation with mite allergen after pretreatment of the tissue with pranlukast , dexamethasone , or both . The P05113 protein level in the culture medium was measured , and in situ hybridization of P05113 and Q9Y271 mRNA was performed using lung tissues . RESULTS : Pretreatment of lung tissues with pranlukast alone significantly decreased the amount of P05113 protein in the culture medium by 40 % . The combination of pranlukast and dexamethasone synergistically enhanced this effect . Quantitative in situ hybridization with image analysis revealed abundant expression of P05113 mRNA in eosinophils , lymphocytes , and mast cells in sensitized and allergen-stimulated lung tissues . Q9Y271 mRNA was detected in macrophages , smooth muscle cells , eosinophils , and mast cells , but was less expressed in lymphocytes . DB01411 -induced inhibition of P05113 mRNA expression was noted in various cells , irrespective of their Q9Y271 mRNA expression status . In addition , cysteinyl leukotrienes per se failed to upregulate the P05113 production . CONCLUSION : Our results indicate that pranlukast inhibits P05113 synthesis via a mechanism distinct from Q9Y271 antagonism . CaMKII autonomy is substrate-dependent and further stimulated by Ca2+/calmodulin . A hallmark feature of Ca(2+)/calmodulin ( P62158 ) -dependent protein kinase II ( CaMKII ) regulation is the generation of Ca(2+)-independent autonomous activity by DB00156 -286 autophosphorylation . CaMKII autonomy has been regarded a form of molecular memory and is indeed important in neuronal plasticity and learning/memory . DB00156 -286-phosphorylated CaMKII is thought to be essentially fully active ( approximately 70-100 % ) , implicating that it is no longer regulated and that its dramatically increased Ca(2+)/ P62158 affinity is of minor functional importance . However , this study shows that autonomy greater than 15-25 % was the exception , not the rule , and required a special mechanism ( T-site binding ; by the T-substrates AC2 or Q13224 ) . Autonomous activity toward regular R-substrates ( including tyrosine hydroxylase and GluR1 ) was significantly further stimulated by Ca(2+)/ P62158 , both in vitro and within cells . Altered K(m) and V(max) made autonomy also substrate- ( and DB00171 ) concentration-dependent , but only over a narrow range , with remarkable stability at physiological concentrations . Such regulation still allows molecular memory of previous Ca(2+) signals , but prevents complete uncoupling from subsequent cellular stimulation . 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 . P48061 inhibits expression of the DB01221 receptor 's Q13224 subunit through a histone deacetylase-dependent pathway contributing to neuronal survival . Homeostatic chemokines , such as P48061 , can affect neuronal activity by the regulation of inhibitory and excitatory neurotransmission , but the mechanisms involved are still undefined . Our previous studies have shown that P48061 protects cortical neurons from excitotoxicity by promoting the function of the gene-repressor protein Rb , which is involved in the recruitment of chromatin modifiers ( such as histone deacetylases ( HDACs ) ) to gene promoters . In neurons , Rb controls activity-dependent genes essential to neuronal plasticity and survival , such as the N-methyl-D-aspartic acid ( DB01221 ) receptor 's subunit Q13224 , the expression of which in the tetrameric ion channel largely affects calcium signaling by glutamate . In this study , we report that P48061 differentially modulates intracellular responses after stimulation of synaptic and extrasynaptic DB01221 receptors , by a specific regulation of the Q13224 gene that involves HDACs . Our results show that P48061 selectively inhibits Q13224 expression in vitro and in vivo altering DB01221 -induced calcium responses associated with neuronal death , while promoting prosurvival pathways that depend on stimulation of synaptic receptors . Along with previous studies , these findings underline the role of P48061 / P61073 in the regulation of crucial components of glutamatergic transmission . These novel effects of P48061 may be involved in the physiological function of the chemokine in both developing and mature brains . Repeated administration of mirtazapine attenuates oxaliplatin-induced mechanical allodynia and spinal Q13224 up-regulation in rats . Chemotherapic drugs may elicit acute or chronic peripheral neuropathies . Mirtazapine , as an antidepressant , is also used for the treatment of neuropathic pain . The current study aimed to investigate the effect of mirtazapine on the oxaliplatin-induced neuropathy in rats as well as the underlying mechanism . A neuropathy model was established in Sprague-Dawley rats by intraperitoneal ( i.p. ) injection of oxaliplatin 4 mg/kg twice a week for 4 weeks . The therapeutic potential of mirtazapine 10 , 20 , and 30 mg/kg/day per-orally for 28 consecutive days was evaluated . Subsequently , a dose of 1 mg/kg of WAY100635 i.p. , a selective antagonist of P08908 receptor , was preadministrated before mirtazapine 20 mg/kg/day per-orally in oxaliplatin-induced neuropathy . The behavioral tests and the expression of DB01221 receptor subunit Q13224 were determined . The results displayed that repeated administration of mirtazapine 20 or 30 mg/kg/day for 28 consecutive days significantly attenuated the mechanical allodynia and the up-regulation of spinal cord Q13224 but not the cold hyperalgesia in rats with oxaliplatin-induced neuropathy , which was reversed by WAY100635 preadministration . Our findings suggest that oxaliplatin-induced mechanical allodynia is associated with spinal Q13224 up-regulation , which may be attenuated by mirtazapine administration . Reduced maximal inhibition in phenotypic susceptibility assays indicates that viral strains resistant to the P51681 antagonist maraviroc utilize inhibitor-bound receptor for entry . DB04835 is a P51681 antagonist in clinical development as one of a new class of antiretrovirals targeting human immunodeficiency virus type 1 ( HIV-1 ) coreceptor binding . We investigated the mechanism of HIV resistance to maraviroc by using in vitro sequential passage and site-directed mutagenesis . Serial passage through increasing maraviroc concentrations failed to select maraviroc-resistant variants from some laboratory-adapted and clinical isolates of HIV-1 . However , high-level resistance to maraviroc was selected from three of six primary isolates passaged in peripheral blood lymphocytes ( PBL ) . The SF162 strain acquired resistance to maraviroc in both treated and control cultures ; all resistant variants were able to use P61073 as a coreceptor . In contrast , maraviroc-resistant virus derived from isolates CC1/85 and RU570 remained P51681 tropic , as evidenced by susceptibility to the P51681 antagonist P35240 -C , resistance to the P61073 antagonist DB06809 , and an inability to replicate in P51681 Delta32/Delta32 PBL . Strain-specific mutations were identified in the V3 loop of maraviroc-resistant CC1/85 and RU570 . The envelope-encoding region of maraviroc-resistant CC1/85 was inserted into an NL4-3 background . This recombinant virus was completely resistant to maraviroc but retained susceptibility to aplaviroc . Reverse mutation of gp120 residues 316 and 323 in the V3 loop ( numbering from HXB2 ) to their original sequence restored wild-type susceptibility to maraviroc , while reversion of either mutation resulted in a partially sensitive virus with reduced maximal inhibition ( plateau ) . The plateaus are consistent with the virus having acquired the ability to utilize maraviroc-bound receptor for entry . This hypothesis was further corroborated by the observation that a high concentration of maraviroc blocks the activity of aplaviroc against maraviroc-resistant virus . 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 . Molecular targeting therapy against promyelocytic leukemia protein using arsenic acids in experimental intracranial medulloblastoma . Our previous study using human Daoy medulloblastoma cells showed that the promyelocytic leukemia ( P29590 ) gene was significantly upregulated ( 2.5-fold ) in cells positive to prominin-1 antigen ( CD133 ) , a possible marker for cancer initiating cells . DB01169 ( As(2)O(3) ) is known to degrade P29590 protein and has been used for the treatment of patients with acute P29590 . In the present study , the effect of P29590 targeting therapy with As(2)O(3) and cytarabine ( DB00987 ) on Daoy medulloblastoma cell proliferation was investigated . Daoy cells were pretreated with As(2)O(3) for 6 weeks . The As(2)O(3)-pretreated Daoy cells were cultured in medium containing DB00987 and cell viability was examined . Next , the As(2)O(3)-pretreated Daoy cells were inoculated into the nude mouse brain and the effect of DB00987 on the tumor size was evaluated . A significant increase in chemosensitivity to DB00987 was observed in the As(2)O(3)-pretreated Daoy cells in both in vitro and in vivo conditions . P29590 and P24385 ( cyclin D1 ) protein expression of Daoy medulloblastoma cells was downregulated by As(2)O(3) treatment . P29590 has been proposed as a novel therapeutic target to eradicate quiescent leukemia-initiating cells , and P29590 -expressing CD133-positive cells are similarly a potential therapeutic target of treatment for medulloblastoma . DB01373 / P62158 -Dependent Signaling for Prepenetration Development in Colletotrichum gloeosporioides . ABSTRACT Colletotrichum gloeosporioides forms a specialized infection structure , an appressorium , for host infection . Contacting hard surface induces appressorium formation in C. gloeosporioides , whereas hydrophobicity of the contact surface does not affect this infection-related differentiation . To determine if the calcium/calmodulin-dependent signaling system is involved in prepenetration morphogenesis in C. gloeosporioides pathogenic on red pepper , effects of calcium chelator ( EGTA ) , phospholipase C inhibitor ( neomycin ) , intracellular calcium modulators ( TMB-8 and methoxy verampamil ) , and calmodulin antagonists ( chloroproma-zine , phenoxy benzamine , and W-7 ) were tested on conidial germination and appressorium formation . Exogenous addition of Ca(2+) , regardless of concentration , augmented conidial germination , while appressorial differentiation decreased at higher concentrations . Inhibition of appressorium formation by EGTA was partly restored by the addition of calcium ionophore A23187 or CaCl(2) . DB01373 channel blockers and calmodulin antagonists specifically reduced appressorium formation at micromolar levels . These results suggest that biochemical processes controlled by the calcium/calmodulin signaling system are involved in the induction of prepenetration morphogenesis in C. gloeosporioides pathogenic on red pepper . Anaplastic oligodendrogliomas with 1p19q codeletion have a proneural gene expression profile . BACKGROUND : In high grade gliomas , 1p19q codeletion and P00533 amplification are mutually exclusive and predictive of dramatically different outcomes . We performed a microarray gene expression study of four high grade gliomas with 1p19q codeletion and nine with P00533 amplification , identified by CGH-array . RESULTS : The two groups of gliomas exhibited very different gene expression profiles and were consistently distinguished by unsupervised clustering analysis . One of the most striking differences was the expression of normal brain genes by oligodendrogliomas with 1p19q codeletion . These gliomas harbored a gene expression profile that partially resembled the gene expression of normal brain samples , whereas gliomas with P00533 amplification expressed many genes in common with glioblastoma cancer stem cells . The differences between the two types of gliomas and the expression of neuronal genes in gliomas with 1p19q codeletion were both validated in an independent series of 16 gliomas using real-time RT-PCR with a set of 22 genes differentiating the two groups of gliomas ( P42330 , Q96SQ7 , P12643 , Q9BQL6 , P14635 , P24941 , P36222 , Q8WZ74 , O43602 , P00533 , Q8IUC8 , P32455 , P18065 , P46940 , L1CAM , P13591 , Q13253 , Q13516 , Q86YL7 , P00750 , Q15063 , Q8IUD6 ) . Immunohistochemical study of the most differentially expressed neuronal gene , alpha-internexin , clearly differentiated the two groups of gliomas , with 1p19q codeletion gliomas showing specific staining in tumor cells . CONCLUSION : These findings provide evidence for neuronal differentiation in oligodendrogliomas with 1p19q codeletion and support the hypothesis that the cell of origin for gliomas with 1p19q codeletion could be a bi-potential progenitor cell , able to give rise to both neurons and oligodendrocytes .

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interacts_with DB04839?

multiple_choice

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 ) . 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 . Loss of P12830 independently predicts the lymph node status in colorectal cancer . BACKGROUND : The identification of biomarkers that improve risk stratification in patients with colorectal cancer ( CRC ) is still a challenge . The objective of our study was to identify independent protein markers as predictors of lymph node ( N ) stage in CRC . METHODS : Tumour specimens from 221 CRC patients were mounted onto a multiple-punch tissue microarray and evaluated for 21 tumour related factors and one host related factor involved in CRC carcinogenesis , namely β-catenin , P12830 , P00533 , pERK , O75330 , pAKT , pSMAD2 , P38936 , p16 , Bcl-2 , Ki-67 , O14727 , MST1 , P30086 , P15692 , EphB2 , P09237 , Laminin5γ2 , P15941 , Q99626 , caspase-3 as well as intra-tumoural and stromal CD8+ tumour infiltrating lymphocytes ( iTILs and sTILs ) . RESULTS : Node positive cancers showed significant losses for P38936 ( p = 0.026 ) , Bcl-2 ( p = 0.027 ) , O14727 ( p = 0.033 ) , EphB2 ( p = 0.006 ) , P12830 ( p < 0.001 ) , P30086 ( p = 0.019 ) , CD8+ iTILs and sTILs ( p < 0.001 and p = 0.008 , respectively ) and cytoplasmic MST1 ( p = 0.014 ) . Based on the area under the receiver operating characteristic curve ( AUC ) EphB2 , P12830 , iTILs and sTILs were identified as potential predictors of N stage ( AUC values > 0.6 ) , but only loss of P12830 was an independent predictor in multivariate analysis . CONCLUSIONS : P12830 appears to be a strong predictor of N stage in CRC and should be considered in pre-operative and post-operative management of colon and rectal cancer patients . Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma : a case report . Uterine carcinosarcoma ( malignant mixed Mullerian tumor ) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells . We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage ( IB due to FIGO ) uterine carcinosarcoma of the homologous type with superficial ( 3mm ) myo-invasion . The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies . Positive immunostaining for steroid receptors ( estrogen-alpha and progesterone receptors ) , cytokeratin , and P00533 was detected only in the carcinomatous area , whereas beta-catenin , BCL-2 , P35354 , p16(INK4a) , P60484 , Q8IUH3 , and vimentin were immunoreactive in both components . P10275 , CD10 , desmin , HER-2/neu , and P04637 were found to be negative either in the carcinomatous or in the sarcomatous area . Tumor proliferative activity was higher in the carcinomatous ( 25 % ) than in the sarcomatous ( 2 % ) component . Based on these findings , immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy . Androgen-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 . 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 . 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 . P10275 -induced tumor suppressor , B2CW77 , inhibits breast cancer growth and transcriptionally activates p53/p73-mediated apoptosis in breast carcinomas . P10275 ( AR ) expression by immunohistochemistry correlates with better prognosis and survival among breast cancer patients . We and others have shown that AR inhibits proliferation and induces apoptosis in breast cancer cells . However , the mechanism of AR 's anti-tumor effect in breast cancer is still not fully understood . Our recent study indicates that AR upregulates expression of tumor suppressor gene P60484 by promoter activation in breast cancer . B2CW77 , encoding B2CW77 protein , is a newly identified gene , which shares a bidirectional promoter with P60484 and is transcribed in the opposite direction . So far , the function of B2CW77 has never been studied in tumorigenesis . Here , we define B2CW77 as a tumor suppressor in breast carcinomas , which inhibits tumor growth and invasiveness . After analyzing 188 normal breast and 1247 malignant breast cancer tissues , we observed the loss of B2CW77 in multiple breast cancer subtypes and this decreased B2CW77 expression associates with tumor progression and increasing histological grade in invasive carcinomas . We characterize B2CW77 , for the first time , as a transcription factor , directly promoting the expression of P04637 and O15350 , with consequent elevated apoptosis and cell cycle arrest in breast cancer cells . We demonstrate , in vitro and in murine xenograph models , that both B2CW77 and P60484 are AR-target genes , mediating androgen-induced growth inhibition and apoptosis in breast cancer cells . Our observations suggest that B2CW77 might be used as a potential prognostic marker and novel therapy target for breast carcinomas . 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 . The genes encoding the human CC-chemokine receptors CC-CKR1 to CC-CKR5 ( P32246 - P51681 ) are clustered in the P38936 .3-p24 region of chromosome 3 . The five human CC-chemokine receptors functionally characterized to date were mapped by using a radiation hybrid panel and YAC contigs . The genes encoding CC-CKR1 , CC-CKR2 , CC- P51677 , and CC-CKR5 ( designated respectively P32246 , P41597 , P51677 , and P51681 in the Genome Data Bank ) were found to be clustered in the 3p21.3 region of chromosome 3 , between the AFM362WB9 and the WI-6983 markers . The four genes fall within a total distance of about 350 kb . The fifth gene ( P51679 , encoding the CC-CKR4 receptor ) was located more distally ( 3p24 ) on the same chromosome , between the FB18G7 and the D3S1768 markers . These localizations were confirmed by mapping the genes into the YAC contigs covering these regions . The clustering of chemokine receptor genes suggests a relatively recent expansion of the gene family by gene duplication . Deletions and duplications of the 3p21 region have been described in neoplastic disorders of the hematopoietic lineage , suggesting a potential link with the CC-chemokine receptor gene family . DB01079 and other serotonergic agents : what is the evidence ? Through effects on gastrointestinal motor and secretory function as well as visceral sensation , serotonin ( 5-HT ) plays a key role in the pathogenesis of irritable bowel syndrome ( IBS ) . In particular , 5- Q9H205 and Q13639 receptors appear to be very important in IBS . This article critically appraises the evidence supporting the use of the 5- Q9H205 receptor antagonist alosetron in the treatment of women with diarrhea-predominant IBS . The safety profile and restricted-use program for alosetron is also reviewed . This discussion is followed by a comprehensive review of the efficacy and safety data in support of tegaserod for women with constipation-predominant IBS . 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 . Regulation of ciliary differentiation of human respiratory epithelial cells by the receptor for hyaluronan-mediated motility on hyaluronan-based biomaterials . Selecting a scaffold that facilitates ciliary differentiation of respiratory epithelial cells ( RECs ) is crucial in developing tissue engineered respiratory epithelium . DB08818 derivative membranes , consisting of an esterified form of hyaluronan ( HYAFF ) , have been proved to promote ciliary differentiation of RECs in the presence of retinoic acid ( RA ) . However , the regulatory mechanism of ciliary differentiation-promoting effect of hyaluronan-based biomaterials remains unknown . In addition to investigating the ciliary differentiation of RECs on HYAFF with/without RA compared with that on collagen with/without RA , this study elucidates the role of the receptor for hyaluronan-mediated motility ( O75330 ) in promoting ciliary differentiation of RECs . Analytical results of culturing RECs on collagen and HYAFF indicate that only HYAFF can increase the ciliary differentiation of RECs under RA-free conditions . The expression level of O75330 mRNA of RECs more significantly decreases on collagen than that on HYAFF without RA . Therefore , by using lentiviral vector-based short hairpin RNA targeting O75330 , the study further reveals that knockdown of O75330 obviously inhibits the ciliary differentiation of RECs on collagen with RA and on HYAFF with/without RA . In addition to demonstrating that hyaluronan-based biomaterials partially " replace " RA in the ciliary differentiation of RECs , which is regulated by O75330 , this study establishes that O75330 regulates the ciliary differentiation-promoting effect of RA on RECs . Production of paired helical filament , tau-like proteins by PC12 cells : a model of neurofibrillary degeneration . Neuron-like cells derived from a rat pheochromocytoma cell line ( PC12 ) and differentiated with nerve growth factor produce a paired helical filament ( PHF ) -like antigen when they are subjected to heat shock ( Wallace et al. : Mol Brain Res 19:149-155 , 1993 ) . It accumulates in a localized region of the perinuclear cytoplasm and reacts with monoclonal antitau antibodies , which identify epitopes in the N- and C-terminal halves and the microtubule-binding domain of tau protein . The observed profile of immunoreactivity suggests the presence of full-length and C-terminally truncated tau in a region of perinuclear cytoplasm in which no structurally intact PHFs could be demonstrated by conventional transmission electron microscopy . The accumulated tau protein colocalized with antibodies raised against mitochondrial outer membrane proteins and was associated with the presence of numerous mitochondrial profiles that were demonstrated with electron microscopy . Because differentiated PC12 cells pretreated with colcemid or DB01229 prior to heat shock fail to exhibit perinuclear PHF-like immunoreactivity , the reported response to heat shock appears to require an intact system of intracellular microtubules . This PC12 system provides a model in which the metabolic and molecular biological underpinnings of neuronal degeneration in Alzheimer 's disease can be manipulated . The system may eventually be applicable to the development of pharmaceutical agents that interfere with formation and/or degeneration of P10636 in Alzheimer 's disease . 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 . High-density SNP haplotyping suggests altered regulation of tau gene expression in progressive supranuclear palsy . Two extended haplotypes exist across the tau gene-H1 and H2-with H1 consistently associated with increased risk of progressive supranuclear palsy ( PSP ) . Using 15 haplotype tagging SNPs ( htSNPs ) , capturing > 95 % of P10636 haplotype diversity , we performed association analysis in a US sample of 274 predominantly pathologically confirmed PSP patients and 424 matched control individuals . We found that PSP risk is associated with one of two major ancestral H1 haplotypes , H1B , increasing from 14 % in control individuals to 22 % in PSP patients ( P < 0.001 ) . In young PSP patients , the H1B risk could be localized to a 22 kb regulatory region in intron 0 ( P < 0.001 ) and could be fully explained by one SNP , htSNP167 , creating a P18428 -1c/ Q12800 /CP2 site , shown to regulate the expression of genes in other neurodegenerative disorders . Luciferase reporter data indicated that the 182 bp conserved regulatory region , in which htSNP167 is located , is transcriptionally active with both alleles differentially influencing expression . Further , we replicated the htSNP167 association in a second , independently ascertained US PSP patient-control sample . However , the htSNP association showed that H1 risk alone could not explain the overall differences in H1 and H2 frequencies in PSP patients and control individuals . Thus , risk variants on different H1 htSNP haplotypes and protective variants on H2 contribute to population risk for PSP . 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 . DNA damage-related gene expression as biomarkers to assess cellular response after gamma irradiation of a human lymphoblastoid cell line . Since defects in molecular mechanisms controlling DNA repair , cell cycle checkpoint and apoptosis could modify cellular sensitivity to DNA damaging agents , we have conducted a multiparametric molecular analysis for better understanding the regulation pathways leading to cell survival or cell death after irradiation . Using a human lymphoblastoid cell line , we have analysed , following gamma irradiation ( 0.5 , 1 , 2 , 4 , 8 , 16 and 32 Gy , at 0.5 , 24 , 48 and 72 h after treatment ) , the correlation between proliferation , cell cycle analysis , apoptosis and micronuclei frequency with the expression of P04637 , P38936 , DNA LIGASE 1 , P12004 , Q07812 , O43927 -2 , Q16611 , P61803 , P42575 -Long and -Short forms mRNAs . We have found that whereas P04637 , Q16611 , P42575 -Short form , and P61803 were expressed at constant levels , P38936 , P12004 , Q07812 were up-regulated , P42575 -Long form , DNA LIGASE 1 , and BCL-2 were down-regulated . These modifications of expression were significantly correlated with doses , survival , proliferation , cell cycle delays , and apoptosis . A positive correlation of P38936 and Q07812 , and a borderline negative correlation with BCL-2 expressions were observed with micronuclei frequency for doses ranging from 0.5 to 4 Gy . In conclusion , our data clearly demonstrate that gene expression profiling , which is easier and more rapid to conduct than the assessments of classical phenotypic responses , could be useful to improve knowledge concerning pathways involved in cellular response to irradiation , knowing that such biomarkers could constitute tools to assess radio-sensitivity/radio-resistance . Oncogene ( 2000 ) 19 , 916 - 923 . Identification of novel genetic alterations in samples of malignant glioma patients . Glioblastoma is the most frequent and malignant human brain tumor . High level of genomic instability detected in glioma cells implies that numerous genetic alterations accumulate during glioma pathogenesis . We investigated alterations in AP-PCR DNA profiles of 30 glioma patients , and detected specific changes in 11 genes not previously associated with this disease : Q86UP9 , Q13326 , Q13639 , P05556 , P31327 , P07225 , P55259 , Q9UJ96 , Q08499 , Q8N743 , and Q14642 . Further correlations revealed that 8 genes might play important role in pathogenesis of glial tumors , while changes in P55259 , Q9UJ96 and Q8N743 should be considered as passenger mutations , consequence of high level of genomic instability . Identified genes have a significant role in signal transduction or cell adhesion , which are important processes for cancer development and progression . According to our results , Q86UP9 might be characteristic of primary glioblastoma , Q13326 , Q13639 , P05556 , P31327 , P07225 and Q14642 were detected predominantly in anaplastic astrocytoma , suggesting their role in progression of secondary glioblastoma , while alterations of Q08499 seem to have important role in development of both glioblastoma subtypes . Some of the identified genes showed significant association with p53 , p16 , and P00533 , but there was no significant correlation between loss of P60484 and any of identified genes . In conclusion our study revealed genetic alterations that were not previously associated with glioma pathogenesis and could be potentially used as molecular markers of different glioblastoma subtypes . 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 . 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 . P10275 expression induces P09038 , FGF-binding protein production , and P09038 release in prostate carcinoma cells : role of P09038 in growth , survival , and androgen receptor down-modulation . BACKGROUND : Alterations in fibroblast growth factors ( FGFs ) production and/or FGF receptors expression have been described to play key roles in prostate tumor progression , particularly in androgen-independent tumors . However , the role of androgen receptor ( AR ) in altering FGF-mediated growth and survival of prostatic neoplastic cells has not been completely defined . In this study , we investigated the alterations in P09038 production and utilization by the PC3 cell line , after transfection with a full-length AR . METHODS : P05230 ,2,7 , FGF-binding protein ( Q14512 ) production and FGF receptor ( FGFR ) 1-4 expression were investigated by polymerase chain reaction ( PCR ) and Western blot analysis . RESULTS : De novo AR expression by PC3 cells restores P21802 IIIb isoform expression and sensitivity to P21781 and P09038 . Androgen stimulation induces AR+ PC3 clones to secrete Q14512 , likely responsible for activation and mobilization from the extracellular matrix of the high amounts of P09038 produced by the same cells . In addition to the effects on cell proliferation , P09038 maintains the survival of AR+ PC3 clones through a positive modulation of the Bcl-2 protein and down-modulates AR protein expression , allowing the escape of selected clones from androgen regulation . CONCLUSION : In the presence of an active AR , the combined production of P09038 and Q14512 may play an important role in the progression of prostate cancer through the selection of AR- clones expressing high levels of Bcl-2 .

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MH_train_1484

interacts_with DB01356?

multiple_choice

Selective inhibition of P01579 -induced autophagy by Mir155- and Mir31-responsive P41221 and SHH signaling . Autophagy is one of the major immune mechanisms engaged to clear intracellular infectious agents . However , several pathogens have evolved strategies to evade autophagy . Here , we demonstrated that Mycobacteria , Shigella , and Listeria but not Klebsiella , Staphylococcus , and Escherichia inhibit P01579 -induced autophagy in macrophages by evoking selective and robust activation of WNT and SHH pathways via P42345 . Utilization of gain- or loss-of-function analyses as well as mir155-null macrophages emphasized the role of P42345 -responsive epigenetic modifications in the induction of Mir155 and Mir31 . Importantly , cellular levels of PP2A , a phosphatase , were regulated by Mir155 and Mir31 to fine-tune autophagy . Diminished expression of PP2A led to inhibition of P49841 , thus facilitating the prolonged activation of WNT and SHH signaling pathways . Sustained WNT and SHH signaling effectuated the expression of anti-inflammatory lipoxygenases , which in tandem inhibited P01579 -induced JAK- P35610 signaling and contributed to evasion of autophagy . Altogether , these results established a role for new host factors and inhibitory mechanisms employed by the pathogens to limit autophagy , which could be targeted for therapeutic interventions . [ Regulation of P04271 expression during long term potentiation ] . In this study , contributions of intracellular regulatory cascades in the induction of P04271 expression in rat hippocampal P00915 area during long term posttetanic potentiation ( LTP ) were estimated . The activation of transcription factor p53 ( positive regulator of P04271 transcription ) by nutlin-3 increased the basal content of P04271 mRNA up to 151 % of the control level , which was significantly lower than its content in tetanized slices ( 280 % ) . Therefore , p53 seems to be not unique transcription factor upregulating P04271 expression during LTP . The inhibitor of Ca2+/calmodulin-dependent kinases ( CaMKs ) KN-93 fully blocked the increase of P04271 mRNA after tetanization , while KN-92 ( inactive analogue of KN-93 ) was ineffective . The inhibitor of CaMKII and receptor tyrosine kinases DB02152 essentially suppressed P04271 expression during LTP , the inhibition of MAPK p38 or P51812 moderately decreased , and the inhibition of Q02750 did not influence P04271 mRNA content . Thus , CaMKs play a key role in the induction of P04271 expression during LTP . 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 . Chromosomal localization of the human prostanoid receptor gene family . Prostaglandins ( PGD2 , DB00917 , PGF2 alpha , and DB01240 ) and thromboxane A2 ( TXA2 ) are biologically active molecules derived from the metabolism of arachidonic acid by cyclooxygenases . They produce a wide variety of physiological and pathophysiological effects mediated through specific G protein-coupled cell surface receptors . In this study , we have mapped the chromosomal positions of the human genes that encode the DB00917 receptor subtypes ( P34995 , PTGER2 , and P43115 ) , the PGF2 alpha receptor ( P43088 ) , the P43119 ( P43119 ) , and the TXA2 receptor ( P21731 ) using in situ hybridization . The P34995 , P21731 , and P43119 genes mapped to chromosome 19 at positions 19p13.1 , 19p13.3 , and 19q13.3 , respectively . The P43088 and P43115 genes mapped to chromosome 1 at positions 1p31.1 and 1p31.2 , respectively , and PTGER2 gene mapped to chromosome band 5p13.1 . Effect of metoclopramide , ondansetron and granisetron on aldosterone secretion in man . The plasma aldosterone response following the administration of drugs with antagonist and agonist activity at Serotonin 3 and 4 ( 5- Q9H205 & 4 ) receptors has been examined in 9 healthy male volunteers receiving the following four treatments i.v. in a randomised , cross-over sequence : ondansetron 8 mg , granisetron 3 mg , metoclopramide 20 mg , and saline 20 ml . DB01233 significantly increased the mean plasma aldosterone level to 196 % of basal level at 5 min . It rose to 234 % at 15 min and remained at more than 185 % of basal level for the duration of the experiment . The response to ondansetron and granisetron did not differ significantly from placebo . If dopamine antagonism is discounted , the results suggest that metoclopramide-induced aldosterone secretion results from its agonist activity at Q13639 receptors , although slow neuronal depolarization via an unidentified receptor remains a possibility . Antagonism at the 5- Q9H205 receptor plays no role , as the selective antagonist , granisetron , did not elicit a significant response . It seems unlikely that the Q13639 receptor is the second , low affinity binding site of ondansetron , unless it had no agonist activity at this receptor . [ The role of glycogen synthase kinase-3 beta in the pathogenesis of liver ischemia reperfusion injury ] . OBJECTIVE : To investigate the role of the key intracellular signaling molecule glycogen synthase kinase-3 beta in the mechanism of liver ischemia reperfusion ( IR ) . METHODS : C57BL/6 mice were subjected to 90 min warm liver cephalad lobe ischemia , followed by various length of reperfusion . Experiment groups included sham control group , liver IRI model group and glycogen synthase kinase-3 beta inhibitor-treated group ( SB216763 in DB01093 , 25 g/kg , i.p , 2 hour prior to the onset of liver ischemia ) . The expression of glycogen synthase kinase-3 beta protein was analysed by Western blotting . The serum ALT levels were determined to reflect the function of liver . The affected liver lobes were harvested for histology analysis . The inflammatory gene expression was detected by Quantitative PCR . RESULTS : By western blot analysis , we found that ischemia itself activated glycogen synthase kinase-3 beta by a significant decrease of its phosphorylation . P49841 inhibitor SB216763-pretreatment ameliorated the liver damages significantly as compared to the controls ( sALT : 2046+/-513 U/L vs 5809+/-1689 U/L , P = 0.0153 ) , and suppressed the gene expressions of IL-12 , TNFa , IL-1b and P05231 . CONCLUSIONS : This study demonstrated that the ischemia process modulated liver innate immune activation via a GSK-3-dependent mechanism which favored the development of a pro-inflammation response and lead to liver tissue damages . GSK-3b may be a new therapeutic target to ameliorate liver IRI in transplant patients . 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 . Glutamate activates c-fos in glial cells via a novel mechanism involving the glutamate receptor subtype mGlu5 and the transcriptional repressor Q9Y2W7 . Activation of c-fos in brain is related to coupling of neuronal activity to gene expression , but also to pathological conditions such as seizures or excitotoxicity-induced cell death . Glutamate activates c-fos in neurons through the calcium-dependent phosphorylation of CREB by P29323 and/or CaMKIV kinase pathways downstream DB01221 -receptors . In glial cells , however , the activation of c-fos by glutamate is poorly understood . Because glial cells actively modulate neuronal excitability and the brain 's response to injury , we studied the mechanisms by which glutamate activates c-fos in rat cortical glial cells . Glutamate potently induced c-fos mRNA in a calcium-dependent manner , as demonstrated by using the calcium chelator BAPTA-AM . Glutamate-induced c-fos mRNA expression was not sensitive to inhibitors of P29323 , p38(MAPK) , or CaMK pathways , indicating that glial c-fos is activated by a distinct mechanism . Thapsigargin abolished the glutamate effect on c-fos mRNA , indicating ER calcium mobilization . Additionally , glutamate induction of c-fos mRNA was sensitive to the P41594 antagonist MPEP but not the DB01221 -R antagonist MK-801 . In luciferase reporter assays , DRE , which actively represses c-fos by binding the calcium-binding transcriptional repressor Q9Y2W7 , was activated by glutamate , whereas SRE and CRE were not . Finally , glutamate caused the nuclear export of Q9Y2W7 in astrocytes , and transfection of astrocytes with a mutant variant of Q9Y2W7 that constitutively binds DNA inhibited glutamate-induced c-Fos expression . These findings are in sharp contrast to the mechanism described in neurons and suggest a novel pathway activated by glutamate in glial cells that employs P41594 , ER calcium , and the derepression of c-fos at the DRE . Silencing of ALA dehydratase affects ALA-photodynamic therapy efficacy in K562 erythroleukemic cells . Synthesis of protoporphyrin IX ( PpIX ) by malignant cells is essential for the success of ALA-based photodynamic therapy ( PDT ) . Two key enzymes that were described as affecting PpIX accumulation during ALA treatment are porphobilinogen deaminase ( P08397 ) and ferrochelatase . Here , we show that down regulation of ALA dehydratase ( P13716 ) expression and activity by specific shRNA induced a marked decrease in PpIX synthesis in K562 erythroleukemic cells . Photo-inactivation efficacy following DB00855 was directly correlated with P13716 -silencing and cellular levels of PpIX . MTT metabolism following DB00855 was shown to be 60 % higher in P13716 -silenced cells in comparison to control cells , indicating that mitochondria were protected in the silenced cells . Morphological analysis by scanning electron microscopy ( SEM ) of cells treated by DB00855 showed no morphological changes in P13716 -silenced cells , in contrast to controls exhibiting cell deformations and lysis . Membrane integrity following DB00855 was kept intact and undamaged in P13716 -silenced cells as examined by P08758 -FITC/PI staining and LDH-L leakage . We conclude that P13716 , although it is present in the cell at abundant levels , has a major and limiting role in regulating PpIX synthesis and DB00855 outcome . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . Gene transcription abnormalities in canine atopic dermatitis and related human eosinophilic allergic diseases . Canine atopic dermatitis ( AD ) is clinically similar to human AD , implicating it as a useful model of human eosinophilic allergic disease . To identify cutaneous gene transcription changes in relatively early inflammation of canine AD , microarrays were used to monitor transcription in normal skin ( n=13 ) and in acute lesional AD ( P13716 ) and nearby visibly nonlesional AD ( NLAD ) skin ( n=13 ) from dogs . Scanning the putative abnormally transcribed genes , several potentially relevant genes , some abnormally transcribed in both NLAD and P13716 ( e.g. P05231 , Q8NET5 , Q9UJ68 , and P43405 ) , were observed . Comparison for abnormally transcribed genes common to two related human diseases , human AD and asthmatic chronic rhinosinusitis with nasal polyps ( aCRSwNP ) , further identified genes or gene sets likely relevant to eosinophilic allergic inflammation . These included : ( 1 ) genes associated with alternatively activated monocyte-derived cells , including members of the monocyte chemotactic protein ( MCP ) gene cluster , ( 2 ) members of the IL1 family gene cluster , ( 3 ) eosinophil-associated seven transmembrane receptor Q14246 and Q9BY15 genes , ( 4 ) interferon-inducible genes , and ( 5 ) keratin genes associated with hair and nail formation . Overall , numerous abnormally transcribed genes were observed only in canine AD ; however , many others are common to related human eosinophilic allergic diseases and represent therapeutic targets testable in dogs with AD . 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 . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . Protein kinase C is involved in Q9BZW8 ( Q9BZW8 ) -mediated cytotoxicity and AP-1 activation in natural killer cells . Q9BZW8 ( Q9BZW8 ) is a member of the P06729 subset of the immunoglobulin superfamily and functions as a triggering molecule on natural killer ( NK ) cells . Previously , we have found that Q9BZW8 -mediated activation of NK cells involves complex interactions involving O43561 , Ras , Raf , P29323 and p38 and that cytolytic function and cytokine production may be regulated by distinct pathways . Here we assessed the role of protein kinase C ( PKC ) in Q9BZW8 -mediated cytotoxicity of YT cells , a human NK cell line . Our data indicate that PKC-delta is activated upon stimulation with monoclonal antibody against Q9BZW8 . Treatment with the PKC inhibitor , bisindolylmaleimide I ( Gö6850 ) , of YT cells or YT cells depleted of Ca2+-dependent isoforms of PKC prior to Q9BZW8 stimulation , resulted in inhibition of natural cytotoxicity and redirected antibody-dependent cellular cytotoxicity . However , inhibition of PKC failed to block Q9BZW8 stimulation of interferon-gamma secretion as opposed to pretreatment with LY294002 , a phosphoinositide 3-kinase inhibitor . We also examined the effect of phorbol 12-myristate 13-acetate ( PMA ) induction on Q9BZW8 gene transcription . PMA induction resulted in a more than two-fold increase of Q9BZW8 transcription . However , when we introduced a three-base substitution mutation to disrupt the activator protein-1 binding site at ( -106 to -100 ) in the Q9BZW8 promoter , we found complete loss of transcriptional activity , including the two-fold increase due to PMA induction of PKC . The present study indicated that PKC may play an important role in Q9BZW8 signalling and activator protein-1 activation . Comparable autoantibody serum levels against amyloid- and inflammation-associated proteins in Parkinson 's disease patients and controls . Naturally occurring autoantibodies ( NAbs ) against a number of potentially disease-associated cellular proteins , including Amyloid-beta1-42 ( Abeta1-42 ) , P37840 ( Asyn ) , myelin basic protein ( MBP ) , myelin oligodendrocyte glycoprotein ( Q16653 ) , and S100 calcium binding protein B ( P04271 ) have been suggested to be associated with neurodegenerative disorders , in particular Alzheimer 's ( AD ) and Parkinson 's disease ( PD ) . Whereas the ( reduced ) occurrence of specific NAbs in AD is widely accepted , previous literature examining the relation of these NAb titres between PD patients and controls , as well as comparing these levels with demographic and clinical parameters in PD patients have produced inconsistent findings . We therefore aimed , in a cross-sectional approach , to determine serum titres of the above NAbs in a cohort of 93 PD patients ( 31 of them demented ) and 194 controls . Levels were correlated with demographic and clinical variables , cerebrospinal fluid Abeta1-42 , total tau and phospho-tau levels , as well as with single nucleotide polymorphisms ( SNPs ) of genes which either have been reported to influence the immune system , the amyloid cascade or the occurrence of PD ( ApoE , P49841 , P01903 , P11021 , P37840 , and Q9UEW8 ) . The investigated NAb titres were neither significantly associated with the occurrence of PD , nor with demographic and clinical parameters , neurodegenerative markers or genetic variables . These results argue against a major potential of blood-borne parameters of the adaptive immune system to serve as trait or state markers in PD . P21731 α promotes tumor growth through an autoregulatory feedback pathway . Tobacco smoking can cause a number of cancers . The role of thromboxane synthase ( TxAS ) in smoking-related cancers is largely unknown . In this study , 37 pairs of tumor and non-tumor lung tissues of non-small-cell lung cancer , 5 lung cancer cell lines , and a mouse tumor model were used to study TxAS and its related molecules . A mouse model of smoking carcinogen 4-methylnitrosamino-1-3-pyridyl-1-butanone ( NNK ) -induced lung tumor showed an increase in TxAS . P21731 ( TP ) was aberrant in lung cancer tissues of smokers . TxAS and TP were increased in lung tissues of NNK-treated mice . The in vitro studies showed that TPα rather than TPβ promoted tumor growth , and NNK increased TPα . NNK-induced TxAS , which depended on activation of cyclooxygenase-2 ( P35354 ) , P29323 and NF-κB , could be inhibited by miR-34b/c . TPα played a positive role in NNK-induced P35354 / P29323 /NF-κB activation , leading to the upregulation of TxAS expression and thromboxane A2 ( TxA2 ) synthesis . The newly synthesized TxA2 could further activate TPα , forming an autoregulatory feedback loop for TPα activation . Collectively , NNK promotes lung tumor growth via inducing TxAS and TPα , which constitutes an auto-positive feedback loop to exaggerate the growth . This study suggests that TPα and TxAS are the ideal targets against smoking-related lung cancer . 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 . Quantification and modeling of tripartite P06729 - , CD58FC chimera (alefacept)- , and CD16-mediated cell adhesion . DB00092 is a chimeric protein combining P19256 immunoglobulin-like domain 1 with human IgG1 Fc . DB00092 mediates adhesion by bridging P06729 on T cells to activating Fc receptors on effector cells , but the equilibrium binding parameters have not been determined . DB00092 mediated T cell killing by NK cells and adhesion between P06729 - and CD16-expressing cells at an optimum concentration of 100 nM . We introduce novel measurements with supported planer bilayers , from which key two-dimensional and three-dimensional parameters can be determined by data fitting . DB00092 competitively inhibited cell bilayer adhesion mediated by the P06729 - P19256 interaction . DB00092 mediated maximal adhesion of P06729 (+) T cells to O75015 , an Fc receptor , in planar bilayers at 500 nM . A mechanistic model for alefacept-mediated cell-bilayer adhesion allowed fitting of the data and determination of two-dimensional binding parameters . These included the density of bonds in the adhesion area , which grew to maintain a consistent average bond density of 200 molecules/microm(2) and two-dimensional association constants of 3.1 and 630 microm(2) for bivalently and monovalently bound forms of alefacept , respectively . The maximum number of CD16 bound and the fit value of 4,350 P06729 per cell are much lower than the 40,000 P06729 per cell measured with anti- P06729 Fab . These results suggest that additional information is needed to correctly predict DB00092 -mediated bridge formation . Role of P15056 in thyroid oncogenesis . P15056 , a cytoplasmic serine-threonine protein kinase , plays a critical role in cell signaling as an activator within the mitogen-activated protein kinase ( MAPK ) pathway . The most common P15056 mutation is the V600E transversion , which causes constitutive kinase activity . This mutation has been found in a multitude of human cancers , including both papillary thyroid cancer ( PTC ) and papillary-derived anaplastic thyroid cancer ( ATC ) , in which it initiates follicular cell transformation . With such a high frequency of P15056 mutations in PTC ( 44 % ) and PTC-derived ATC ( 24 % ) , research in P15056 (V600E) detection for diagnostic purposes has shown high sensitivity and specificity for tumor cell presence . P15056 (V600E) in PTC has also provided valuable prognostic information , as its presence has been correlated with more aggressive and iodine-resistant phenotypes . Such findings have initiated research in targeting oncogenic P15056 in cancer therapeutics . Although multiple phase II clinical trials in patients with iodine-refractory metastatic PTC have shown significant efficacy for sorafenib , a first-generation P15056 inhibitor , the mechanism by which it mediates its effect remains unclear because of multiple additional kinase targets of sorafenib . Additionally , preclinical and clinical studies investigating combination therapy with agents such as selective ( PLX 4032 ) and potent ( DB08896 and ARQ 736 ) small-molecule P15056 inhibitors and Q96HU1 /extracellular signal-regulated kinase ( P29323 ) kinase inhibitors ( AZD6244 ) hold great promise in the treatment of P15056 (V600E) cancers and may eventually play a powerful role in changing the clinical course of PTC and ATC . 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 .

MH_train_1485

MH_train_1485

MH_train_1485

interacts_with DB00559?

multiple_choice

Vitamin D and the regulation of placental inflammation . The vitamin D-activating enzyme 1α-hydroxylase ( O15528 ) and vitamin D receptor ( P11473 ) support anti-inflammatory responses to vitamin D in many tissues . Given the high basal expression of O15528 and P11473 in trophoblastic cells from the placenta , we hypothesized that anti-inflammatory effects of vitamin D may be particularly important in this organ . Pregnant wild type ( WT ) mice i.p. injected with LPS showed elevated expression of mouse Cyp27b1 ( 4-fold ) and P11473 ( 6-fold ) . Similar results were also obtained after ex vivo treatment of WT placentas with LPS . To assess the functional impact of this , we carried out ex vivo studies using placentas -/- for fetal ( trophoblastic ) Cyp27b1 or P11473 . Vehicle-treated -/- placentas showed increased expression of IFN-γ and decreased expression of P22301 relative to +/+ placentas . LPS-treated -/- placentas showed increased expression of O60603 , IFN-γ , and P05231 . Array analyses identified other inflammatory factors that are dysregulated in Cyp27b1(-/-) versus Cyp27b1(+/+) placentas after LPS challenge . Data highlighted enhanced expression of P05112 , P40933 , and Q14116 , as well as several chemokines and their receptors , in Cyp27b1(-/-) placentas . Similar results for P05231 expression were observed with placentas -/- for trophoblastic P11473 . Finally , ex vivo treatment of WT placentas with the substrate for Cyp27b1 , 25-hydroxyvitamin D(3) , suppressed LPS-induced expression of P05231 and the chemokine Ccl11 . These data indicate that fetal ( trophoblastic ) vitamin D plays a pivotal role in controlling placental inflammation . In humans , this may be a key factor in placental responses to infection and associated adverse outcomes of pregnancy . Age-dependent (+) DB01454 -mediated neurotoxicity in mice . In the present study the effects of a neurotoxic regimen of (+)- DB01454 ( 20 mg/kgx4 , s.c. ) in 4- and 10-week-old C57Bl/6J mice during treatment and 7 days post-treatment were examined . Rectal temperatures monitored between (+)- DB01454 injections ( 30 min post-injection/2 h intervals ) revealed hyperthermic responses in both age groups , with the magnitude of the response significantly greater in older mice . Seven days post-treatment , immunoblot analyses of the vesicular monoamine transporter 2 ( Q05940 ) , and tyrosine hydroxylase ( TH ) revealed significant reductions ( -37 and -58 % , respectively ) in the older animals , but not in the younger group , compared to age-matched controls . Dopamine transporter ( Q01959 ) expression was significantly reduced in both 4- and 10-week-old animals ( 26 and 69.7 % , respectively ) . (+)- DB01454 -treated animals also exhibited significantly lower levels of striatal dopamine , and 3,4-dihydroxyphenylacetic acid than controls , again the effect being more pronounced in the older animals . Although both age groups showed evidence of (+)- DB01454 -induced toxicity , our data revealed that older animals exhibited a greater hyperthermic response to (+)- DB01454 and were also are more susceptible to subsequent dopaminergic damage than the younger animals . [ DB02424 administration reduces the number of P07900 -positive germ cells in the mouse embryo : preliminary results ] . 5 mg of DB02424 , an inhibitor of stress protein P07900 which express on mammalian germ cells , were administered to E8 pregnant mice . E17 embryos were removed , and a quantitative analysis of HSP90-immunoreactive cells in the gonad was performed , in comparison to control embryos . First , we observed that the number of germ cells is lower in male than in female embryos , as well in control and experimental embryos . External features of experimental and control embryos did not display any difference . Embryos exposed to geldanamycin exhibit a significant decrease of immunoreactive germ cells . In two embryos , we observed a group of ectopic immunoreactive cells in the pelvic area . We conclude that geldanamycin inhibits germ cells migration , and suggest that this inhibition can lead to ectopic germ cell populations , similar to teratomas . Mycobacteria-primed macrophages and dendritic cells induce an up-regulation of complement C5a anaphylatoxin receptor ( CD88 ) in CD3+ murine T cells . Complement C5a anaphylatoxin is a potent activator of macrophages , neutrophils , and dendritic cells ( DC ) and binds the C5a receptor ( P21730 ; CD88 ) . Although C5a is chemotactic for T cells , expression of P21730 on murine T cells has been disputed . We report here that naïve , Con A-activated , and cytokine ( IL-12 , Q14116 ) -stimulated murine CD3+ T cells from three strains of mice [ C57Bl/6 , B10.nSn ( P01031 +/+ ) , B10.on ( P01031 -/- ) ] lacked P21730 , as evaluated by immunophenotyping with an anti- P21730 mAb . Ligation of CD3 induced a modest up-regulation with 3 % of CD3+ T cells expressing cell surface P21730 . T cells primed by P25054 differentiate into effector T cells . Activation of mycobacteria [ bacillus Calmette-Guerin ( BCG ) ] -sensitized T cells through MHC II and TCR interactions via BCG-infected macrophages enhanced the expression of P21730 with approximately 14 % of CD3+ T cells positive for P21730 . Comparable expression was found in P01031 +/+ as well as P01031 -/- strains of mice ( 14 % and 15 % , respectively ) . Furthermore , anti-CD3-activated T cells were primed by BCG-infected DC , and a larger proportion of the primed T cells expressed P21730 ( 30-40 % ) . Finally , mice infected with BCG showed significant numbers of CD3+ T cells expressing P21730 in the spleens during infection . As P25054 , such as macrophages and DC , can secrete P01031 and cleave P01031 to C5a and C5b through a peptidase , we suggest that macrophage and DC-T cell interactions can up-regulate P21730 on T cells through MHC II-TCR and provide a C5a peptide for additional local activation of T cells via P21730 . 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 . 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 . The use of antibody to complement protein P01031 for salvage treatment of severe antibody-mediated rejection . Desensitized patients are at high risk of developing acute antibody-mediated rejection ( AMR ) . In most cases , the rejection episodes are mild and respond to a short course of plasmapheresis ( PP ) / low-dose DB00028 treatment . However , a subset of patients experience severe AMR associated with sudden onset oliguria . We previously described the utility of emergent splenectomy in rescuing allografts in patients with this type of severe AMR . However , not all patients are good candidates for splenectomy . Here we present a single case in which eculizumab , a complement protein P01031 antibody that inhibits the formation of the membrane attack complex ( MAC ) , was used combined with PP/ DB00028 to salvage a kidney undergoing severe AMR . We show a marked decrease in C5b- P02748 ( MAC ) complex deposition in the kidney after the administration of eculizumab . ADP receptors -- targets for developing antithrombotic agents . Platelet P2 receptors -- P47900 , Q9H244 , and P51575 -- constitute the means by which adenine nucleotides can activate platelets . Coactivation of the Galphaq-coupled P47900 and Galphai2-coupled Q9H244 receptors is necessary for ADP-mediated platelet activation , which forms the basis of using P2 antagonists as antithrombotic drugs . P47900 receptor antagonists inhibit platelet activation , while P47900 knockout mice show longer bleeding times than normal mice but few other problems ; however , its ubiquitous expression in other tissues renders P47900 questionable as an antithrombotic target . The Q9H244 receptor is expressed nearly exclusively in platelets and brain , making it an attractive antithrombotic target . Antagonists for the Q9H244 receptor have been developed that either require metabolic activation to covalently inhibit Q9H244 and are irreversible , or simply are competitive in nature and thus reversible . DB00208 and clopidogrel are irreversible Q9H244 antagonists and have been repeatedly proven as clinical antithrombotic agents . In addition , a recently reported Q9H244 antagonist , CS-747 , shows promise as a future antithrombotic drug . The AR-C series of compounds represent reversible Q9H244 antagonists and have been used extensively to characterize the function of Q9H244 in platelets . Clinical studies show that AR-C69931MX is as effective as clopidogrel ; furthermore , the combination of AR-C69931MX ( cangrelor ) and clopidogrel confers greater antagonism of Q9H244 than either antagonist alone . The P51575 receptor is a calcium channel that functions to potentiate agonist-induced platelet shape change , and its inhibition or loss has little if any effect on hemostasis . A combination of P47900 and Q9H244 antagonists may represent an additional course of antithrombotic treatment . DB00559 , an endothelin receptor antagonist , ameliorates collagen-induced arthritis : the role of P01375 -α in the induction of endothelin system genes . OBJECTIVE : Endothelins ( ETs ) are involved in several inflammatory events . The present study investigated the efficacy of DB00559 , a dual P25101 /ETB receptor antagonist , in collagen-induced arthritis ( CIA ) in mice . TREATMENT : CIA was induced in DBA/1J mice . Arthritic mice were treated with DB00559 ( 100 mg/kg ) once a day , starting from the day when arthritis was clinically detectable . METHODS : CIA progression was assessed by measurements of visual clinical score , paw swelling and hypernociception . Histological changes , neutrophil infiltration and pro-inflammatory cytokines were evaluated in the joints . Gene expression in the lymph nodes of arthritic mice was evaluated by microarray technology . PreproET-1 mRNA expression in the lymph nodes of mice and in peripheral blood mononuclear cells ( PBMCs ) was evaluated by real-time PCR . The differences were evaluated by one-way Q9UNW9 or Student 's t test . RESULTS : Oral treatment with DB00559 markedly ameliorated the clinical aspects of CIA ( visual clinical score , paw swelling and hyperalgesia ) . DB00559 treatment also reduced joint damage , leukocyte infiltration and pro-inflammatory cytokine levels ( IL-1β , TNFα and Q16552 ) in the joint tissues . Changes in gene expression in the lymph nodes of arthritic mice returned to the levels of the control mice after DB00559 treatment . PreproET mRNA expression increased in PBMCs from rheumatoid arthritis ( RA ) patients but returned to basal level in PBMCs from patients under anti- P01375 therapy . In-vitro treatment of PBMCs with TNFα upregulated ET system genes . CONCLUSION : These findings indicate that ET receptor antagonists , such as DB00559 , might be useful in controlling RA . Moreover , it seems that ET mediation of arthritis is triggered by TNFα . 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 . Structural studies with inhibitors of the cell cycle regulatory kinase cyclin-dependent protein kinase 2 . Components of the cell cycle machinery are frequently altered in cancer . Many of these alterations affect the cyclin-dependent kinases ( CDKs ) and their regulation . DB02010 and 7-hydroxystaurosporine ( P55089 -01 ) are two natural product kinase inhibitors originally identified as potent protein kinase C inhibitors . DB02010 is non-selective and too toxic for use in therapy , but P55089 -01 shows greater selectivity , and is in clinical trials . We have determined the crystal structures of staurosporine bound to monomeric P24941 and P55089 -01 bound to active phospho- P24941 /cyclin A . Both compounds mimic the hydrogen bonds made by the adenine moiety of DB00171 , and both exploit the non-polar nature of the adenine-binding site . In the complex with P55089 -01 , a hydrogen-bonded water molecule is incorporated into the non-polar cavity , which provides a partial polar character in the environment of the 7-hydroxyl group . Comparison of the DB00171 -binding site of P24941 with that of other kinases reveals that in Chk1 kinase , a major target for P55089 -01 in the cell , one of the surrounding residues , Ala144 in P24941 , is a serine in Chk1 , thus providing a possible explanation for the effectiveness of P55089 -01 against this kinase . For cells to exit mitosis , the CDKs must be completely inactivated , firstly by the ubiquintin-mediated destruction of the cyclins , followed by dephosphorylation of phospho-Thr160 ( in P24941 ) catalysed by the kinase-associated phosphatase and protein phosphatase 2C . We describe the structure of phospho- P24941 in complex with kinase-associated phosphatase , and discuss the substrate recognition promoted by interactions that are remote from the catalytic site . Genome-wide association study on plasma levels of midregional-proadrenomedullin and C-terminal-pro-endothelin-1 . P05305 ( ET-1 ) and adrenomedullin ( P35318 ) are circulating vasoactive peptides involved in vascular homeostasis and endothelial function . Elevated levels of plasma ET-1 and P35318 , and their biologically stable surrogates , C-terminal-pro-endothelin-1 ( CT-pro-ET-1 ) and midregional proadrenomedullin ( MR-pro- P35318 ) , are predictors of cardiac death and heart failure . We studied the association of common genetic variation with MR-pro- P35318 and CT-pro-ET-1 by genome-wide association analyses in 3444 participants of European ancestry . We performed follow-up genotyping of single nucleotide polymorphisms ( SNPs ) that showed suggestive or significant association in the discovery stage in additional 3230 participants . The minor variants in P03952 ( rs4253238 ) and P00748 ( rs2731672 ) , both part of the kallikrein-kinin system , were associated with higher MR-pro- P35318 ( P=4.46E-52 and P=5.90E-24 , respectively ) and higher CT-pro-ET-1 levels ( P=1.23E-122 and P=1.26E-67 , respectively ) . Epistasis analyses showed a significant interaction between the sentinel SNP of P00748 and P03952 for both traits . In addition , a variant near the P35318 gene ( rs2957692 ) was associated with MR-pro- P35318 ( P=1.05E-12 ) and a variant in P10153 -1 ( rs5370 ) was associated with CT-pro-ET-1 ( P=1.49E-27 ) . The total phenotypic variation explained by the genetic variants was 7.2 % for MR-pro- P35318 and 14.6 % for CT-pro-ET-1 . P03952 encodes plasma kallikrein , a proteolytic enzyme known to cleave high-molecular-weight kininogen to bradykinin and prorenin to renin . We cloned the precursors of P35318 and ET-1 and demonstrated that purified plasma kallikrein can cleave these recombinant proteins into multiple smaller peptides . The discovery of genetic variants in the kallikrein-kinin system and in the genes encoding pre-pro-ET-1 and pre-pro- P35318 provides novel insights into the (co-)regulation of these vasoactive peptides in the vascular system . Characterization of endothelin receptors in the human pulmonary vasculature using DB00559 , SB209670 , and 97-139 . P05305 ( ET-1 ) is believed to have a role in the pathogenesis of pulmonary hypertension , and ET antagonists may therefore be useful in the treatment of the disease . Here we have characterized ET receptors and ligands in human pulmonary tissues . Autoradiography showed P25101 receptors located in resistance and conduit arteries and ETB receptors present in airway smooth muscle . Competition binding studies in human pulmonary artery ( Q9Y251 ) showed a predominance of the P25101 subtype ( 90 % ) . DB00559 ( Ro470203 ) ( Kd P25101 = 12.5 nM , Kd ETB = 1.1 microM ) , SB209670 ( Kd P25101 = 14.3 nM , Kd ETB = 5.0 microM ) , and 97-139 ( Kd P25101 = 5.3 nM , Kd ETB = 19.6 microM ) labeled P25101 receptors with higher affinity than BMS182874 ( 50 % [125I]ET-1 inhibition at 1 microM ) . Sarafotoxin S6c labeled ETB receptors with high affinity ( Kd P25101 = 0.16 microM , Kd ETB = 2.7 nM ) , whereas BQ788 competed with low affinity for [125I]ET-1 binding sites ( Kd = 1.0 microM ) . This study indicates that an P25101 -selective antagonist may be useful in reversing vasoconstriction associated with pulmonary hypertension without affecting ETB-mediated contractile effects in airway smooth muscle or ETB-mediated release of endothelium-derived vasodilators . 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 . Abnormal NF-kappaB signaling pathway with enhanced susceptibility to apoptosis in immortalized keratinocytes . The transcriptional activation and proper regulation of NF-kappaB is known to be important to the apoptotic resistant phenotype of epidermal-derived keratinocytes . By comparing and contrasting the responses of normal foreskin-derived keratinocytes versus an immortalized skin-derived keratinocyte cell line ( i.e. HaCaT cells ) , several molecular defects involving NF-kappaB signaling pathway were delineated in the immortalized keratinocytes . While exposure to P01579 plus TPA produces growth arrest in both normal and immortalized keratinocytes , with rapid phosphorylation of MEKKI and recruitment of distinctive protein kinase C isoforms into the signalosome complex , subsequent molecular events necessary for NF-kappaB activation were abnormal in HaCaT cells . This disrupted NF-kappaB activation in HaCaT cells was accompanied by enhanced susceptibility to UV-light induced apoptosis , which was associated with elevated levels of Q01094 and decreased Q13077 /TRAF2 levels . Additional defects in HaCaT cells included markedly diminished levels of IKKbeta ( and lack of induction of kinase activity ) in response to inflammatory stimuli , a failure of P38936 ( P38936 /CIP1) to associate with P24941 , and a decreased association between p65 and p300 . These studies suggest caution in using HaCaT cells as a substitute for normal keratinocytes to study apoptosis in the skin . Thus , it appears that while the immortalized cells can escape cell cycle checkpoints by elevated levels of Q01094 , an adverse biological consequence of such dysregulated cell cycle control is the inability to activate the anti-apoptotic NF-kappaB signaling pathway . Therefore , exploiting this apoptosis vulnerability in pre-malignant , or immortalized cells , prior to acquiring a death-defying phenotype characteristic of more advanced malignant cell types , provides the basis for an early interventional therapeutic strategy for cutaneous oncologists . 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 . Endothelin receptor antagonist exacerbates autoimmune myocarditis in mice . AIMS : Myocarditis and subsequent dilated cardiomyopathy are major causes of heart failure in young adults . Experimental autoimmune myocarditis ( EAM ) is a mouse model of post-infectious myocarditis and inflammatory cardiomyopathy . The pathological role of endothelin ( ET ) in myocarditis has not been elucidated . MAIN METHODS : EAM was induced by immunization of cardiac myosin peptide with complete Freund 's adjuvant on days 0 and 7 in BALB/c mice . An P25101 /ETB dual receptor antagonist , SB209670 , was administered by a continuous infusion from a subcutaneous pump for 2 weeks . KEY FINDINGS : An increase in the heart-to-body weight ratio was observed in SB209670-treated mice compared with vehicle-treated mice . Heart pathology in SB209670-treated mice was remarkable for gross inflammatory infiltration , in contrast to the lesser inflammation in the hearts of vehicle-treated mice . We found that an ET blockade decreased the number of Foxp3(+) regulatory T cells in the heart . The ET blockade also inhibited the expression of the suppressor of cytokine signaling 3 that plays a key role in the negative regulation of both Toll-like receptor- and cytokine receptor-mediated signaling . EAM is a P01730 (+) T cell-mediated disease . P01730 (+) T cells isolated from SB209670-treated EAM mice produced less P22301 and more inflammatory cytokines , P05231 and Q16552 , than those isolated from vehicle-treated mice . SIGNIFICANCE : The ET receptor antagonist exacerbated autoimmune myocarditis in mice . Our novel findings suggest that ET may play an important role in the regulation of inflammation in myocarditis . DB00171 induces Q99572 receptor-independent cytokine and chemokine expression through P51575 and P56373 receptors in murine mast cells . Extracellular DB00171 mediates a diverse array of biological responses in many cell types and tissues , including immune cells . We have demonstrated that DB00171 induces purinergic receptor P2X(7) mediated membrane permeabilization , apoptosis , and cytokine expression in murine mast cells ( MCs ) . Here , we report that MCs deficient in the expression of the P2X(7) receptor are resistant to the DB00171 -induced membrane permeabilization and apoptosis . However , DB00171 affects the tyrosine phosphorylation pattern of P2X(7)knockout cells , leading to the activation of P27361 /2 . Furthermore , DB00171 induces expression of several cytokines and chemokines in these cells , including P05112 , P05231 , P01579 , P01375 , RANTES , and MIP-2 , at the mRNA level . In addition , the release of P05231 and P35225 to cell-conditioned medium was confirmed by ELISA . The ligand selectivity and pharmacological profile indicate the involvement of two P2X family receptors , P2X(1) and P2X(3) . Thus , depending on genetic background , particular tissue microenvironment , and DB00171 concentration , MCs can presumably engage different P2X receptor subtypes , which may result in functionally distinct biological responses to extracellular nucleotides . This finding highlights a novel level of complexity in the sophisticated biology of MCs and may facilitate the development of new therapeutic approaches to modulate MC activities . Acute pharmacodynamic and antivascular effects of the vascular endothelial growth factor signaling inhibitor DB04849 in Calu-6 human lung tumor xenografts . The vascular endothelial growth factor-A ( P15692 ) signaling pathway , a key stimulant of solid tumor vascularization , is primarily dependent on the activation of the endothelial cell surface receptor P15692 receptor-2 ( P35968 ) . DB04849 is an oral , highly potent small-molecule inhibitor of VEGFR tyrosine kinase activity that inhibits angiogenesis and the growth of human tumor xenografts in vivo . Here , we show pharmacodynamic changes in P35968 phosphorylation induced by DB04849 . In mouse lung tissue , a single dose of DB04849 at 6 mg/kg inhibited P15692 -stimulated P35968 phosphorylation by 87 % at 2 h with significant inhibition ( > or=60 % ) maintained to 24 h . To examine inhibition of P35968 phosphorylation in tumor vasculature by immunohistochemistry , a comprehensive assessment of antibodies to various phosphorylation sites on the receptor was undertaken . Antibodies to the phosphotyrosine epitopes pY1175/1173 and pY1214/1212 were found suitable for this application . Calu-6 human lung tumor xenografts , from mice receiving DB04849 or vehicle treatment ( p.o. , once daily ) , were examined by immunohistochemistry . A significant reduction in tumor vessel staining of phosphorylated P35968 ( pVEGFR-2 ) was evident within 28 h of DB04849 treatment ( 6 mg/kg ) . This effect preceded a significant reduction in tumor microvessel density , which was detectable following 52 h of DB04849 treatment . These data show that DB04849 is a potent inhibitor of P35968 activation in vivo and suggest that DB04849 delivers therapeutic benefit in Calu-6 tumors by targeting vessels dependent on P35968 signaling for survival . In addition , this work highlights the utility of measuring either pY1175/1173 or pY1214/1212 on P35968 as a pharmacodynamic marker of P35968 activation . 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 . 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 . Systemic sclerosis - a systematic overview : part 2 - immunosuppression , treatment of SSc-associated vasculopathy , and treatment of pulmonary arterial hypertension . Here we give an overview over treatment recommendations propagated by the European League Against Rheumatism ( EULAR ) , EULAR Scleroderma Trials and Research Group , the German Network for Systemic Sclerosis , the European Respiratory Society , and the International Society of Heart and Lung Transplantation . As response to immunosuppressant ( IS ) therapy is usually weaker in systematic sclerosis ( SSc ) compared to other connective tissue disorders IS should be considered with caution . To prevent scleroderma renal crisis steroid doses should not exceed 15 mg/d . The definitive role of a number of new immunosuppressant drugs and the effects of autologous stem cell transplantation in systemic clerosis ( SSc ) have to be elucidated . Prostanoids , especially iloprost , are widely used as intravenous formulas for the treatment of severe Raynaud 's phenomenon ( RP ) and digital ulcers ( DU ) . DB01373 antagonists are of limited therapeutic value . DB00559 , an oral endothelin receptor antagonists ( P25101 ) , was shown to prevent new DU , but failed to heal existing DU , while the oral phopshodiesterase inhibitor ( P07237 ) DB00203 reduces the occurrence of RP and might be effective in ulcer healing . Combination therapies of P07237 with P25101 are currently evaluated . Therapy of pulmonary arterial hypertension ( PAH ) is usually started as oral monotherapy , frequently using an P25101 . When this first-line therapy is not tolerated P25101 is substituted by P07237 . If treatment goals are not reached with monotherapy combinationtherapy is started , for example by adding a P07237 to an existing P25101 . In general , treatment of PAH in patients with connective tissue disease follows the same algorithms as in idiopathic PAH . Integrative quantitative proteomics unveils proteostasis imbalance in human hepatocellular carcinoma developed on nonfibrotic livers . Proteomics-based clinical studies represent promising resources for the discovery of novel biomarkers or for unraveling molecular mechanisms underlying particular diseases . Here , we present a discovery study of hepatocellular carcinoma developed on nonfibrotic liver ( nfHCC ) that combines complementary quantitative iTRAQ-based proteomics and phosphoproteomics approaches . Using both approaches , we compared a set of 24 samples ( 18 nfHCC versus six nontumor liver tissue ) . We identified 43 proteins ( 67 peptides ) differentially expressed and 32 peptides differentially phosphorylated between the experimental groups . The functional analysis of the two data sets pointed toward the deregulation of a protein homeostasis ( proteostasis ) network including the up-regulation of the Endoplasmic Reticulum ( ER ) resident P11021 , P14625 , Q15084 , and P07237 and of the cytosolic P0DMV9 , P07900 , P38646 , P0CG48 , Q96KP4 , P10599 , and P55072 as well as the increased phosphorylation of the ER resident calnexin at Ser583 . Antibody-based validation approaches ( immunohistochemistry , immunoblot , Alphascreen(®) , and AMMP(®) ) on independent nfHCC tumor sets ( up to 77 samples ) confirmed these observations , thereby indicating a common mechanism occurring in nfHCC tumors . Based on these results we propose that adaptation to proteostasis imbalance in nfHCC tumors might confer selective advantages to those tumors . As such , this model could provide an additional therapeutic opportunity for those tumors arising on normal liver by targeting the tumor proteostasis network . Data are available via ProteomeXchange with identifier PXD001253 .

MH_train_1486

MH_train_1486

MH_train_1486

interacts_with DB00834?

multiple_choice

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 . Quantitative analysis of the effect of Helicobacter pylori on the expressions of P48431 , Q99626 , Q02817 , P98088 , Q6W4X9 , P04155 , Q03403 , and Q07654 mRNAs in human gastric carcinoma cells . OBJECTIVE : To investigate the phenotypic characters of carcinoma cells and the response of gastric epithelial cells to Helicobacter pylori ( H. pylori ) infection using the gastric carcinoma cell lines . MATERIAL AND METHODS : Real-time reverse transcription-polymerase chain reaction ( RT-PCR ) was used to assess the effect of H. pylori infection on mRNA levels of transcription factors ( P48431 and Q99626 ) , mucin core proteins ( Q02817 , P98088 , and Q6W4X9 ) , and trefoil factor family peptides ( TFF ) ( P04155 , Q03403 , and Q07654 ) in gastric carcinoma cells ( AGS , MKN45 , and KATO III cells ) . H. pylori ATCC 43504 and its isogenic cag pathogenicity island ( P05121 ) deleted mutant were used . RESULTS : These cell lines expressed mixed gastric and intestinal phenotypes . The intestinal phenotype predominated in AGS cells and gastric phenotypes in MKN45 and KATO III cells . In all three cell lines , H. pylori infection inhibited P48431 mRNA expression , but induced the three TFFs mRNAs . In AGS cells , H. pylori induced cag P05121 -dependent mRNA expression of Q99626 , Q02817 , P98088 , and Q6W4X9 . mRNA expressions of Q99626 , P98088 , and Q6W4X9 were inhibited in KATO III cells , whereas Q02817 mRNA expression was unchanged . In MKN45 cells , H. pylori induced the three MUCs mRNAs but inhibited Q99626 mRNA expression . CONCLUSIONS : This study provides a useful platform for selecting appropriate cell lines to model H. pylori-related changes in the gastric epithelium that mirror the changes seen in vivo . The outcome of H. pylori infection may reflect changes in the mucus gel layer caused by altered expression of mucins and TFF peptides . Evaluation of platelet activation , coagulation , and fibrinolytic activation in patients with symptomatic lacunar stroke . BACKGROUND : It is unclear whether hemostasis plays a role in the pathogenesis of ischemic stroke subtypes . OBJECTIVE : We aimed to investigate the possible relationship between different hemostatic markers and lacunar stroke . RESULTS : The study consisted of 30 patients with symptomatic lacunar stroke and 30 healthy age-matched healthy individuals . We analyzed the values of " Mean Platelet Volume , " D-dimer , " soluble p-selectin , " " P00747 Activator Inhibitor Type-1 " ( P05121 ) , " Thrombin-Activatable DB06692 " ( Q96IY4 ) , and " Platelet Factor 4 " ( P02776 ) in patients with lacunar infarct and compared these values to those of control individuals . There were significant differences for D-dimer , mean platelet volume , thrombin-activatable fibrinolysis inhibitor , and platelet factor 4 values in symptomatic lacunar stroke group compared with the control group ( P < 0.01 ) . CONCLUSIONS : Different hemostatic factors may play a role in the pathogenesis of lacunar stroke . Evaluating the role of hemostatic factors on different types of strokes may help us identify new therapeutic strategies and different prognostic stratifications for ischemic stroke . Insights into antifolate resistance from malarial P00374 -TS structures . Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) is an important target of antimalarial drugs . The efficacy of this class of P00374 -inhibitor drugs is now compromised because of mutations that prevent drug binding yet retain enzyme activity . The crystal structures of PfDHFR-TS from the wild type ( TM4/8.2 ) and the quadruple drug-resistant mutant ( V1/S ) strains , in complex with a potent inhibitor WR99210 , as well as the resistant double mutant ( P04264 P21554 ) with the antimalarial pyrimethamine , reveal features for overcoming resistance . In contrast to pyrimethamine , the flexible side chain of WR99210 can adopt a conformation that fits well in the active site , thereby contributing to binding . The single-chain bifunctional PfDHFR-TS has a helical insert between the P00374 and TS domains that is involved in dimerization and domain organization . Moreover , positively charged grooves on the surface of the dimer suggest a function in channeling of substrate from TS to P00374 active sites . These features provide possible approaches for the design of new drugs to overcome antifolate resistance . 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 . 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 . 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 and histone deacetylase-2 mediate dexamethasone-induced repression of P98088 gene expression . Airway occlusion in obstructive airway diseases is caused in part by the overproduction of secretory mucin glycoproteins through the up-regulation of mucin ( MUC ) genes by inflammatory mediators . Some pharmacological agents , including the glucocorticoid dexamethasone ( DB00514 ) , repress mucin concentrations in lung epithelial cancer cells . Here , we show that DB00514 reduces the expression of P98088 , a major airway mucin gene , in primary differentiated normal human bronchial epithelial ( NHBE ) cells in a dose-dependent and time-dependent manner , and that the DB00514 -induced repression is mediated by the glucocorticoid receptor ( GR ) and two glucocorticoid response elements ( GREs ) in the P98088 promoter . The pre-exposure of cells to DB00834 , a GR antagonist , and mutations in either the GRE3 or GRE5 cis-sites abolished the DB00514 -induced repression . Chromatin immunoprecipitation ( ChIP ) assays showed a rapid temporal recruitment of GR to the GRE3 and GRE5 cis-elements in the P98088 promoter in NHBE and in A549 cells . Immunofluorescence showed nuclear colocalization of GR and histone deacetylase-2 ( Q92769 ) in P98088 -expressing NHBE cells . ChIP also showed a rapid temporal recruitment of Q92769 to the GRE3 and GRE5 cis-elements in the P98088 promoter in both cell types . The knockdown of Q92769 by Q92769 -specific short interfering RNA prevented the DB00514 -induced repression of P98088 in NHBE and A549 cells . These data demonstrate that GR and Q92769 are recruited to the GRE3 and GRE5 cis-sites in the P98088 promoter and mediate the DB00514 -induced cis repression of P98088 gene expression . A better understanding of the mechanisms whereby glucocorticoids repress P98088 gene expression may be useful in formulating therapeutic interventions in chronic lung diseases . P00747 binding sites in normal human skin . P00747 is detected in the basal cell layer of the epidermis , keratinocytes can generate plasminogen activators and it is suggested that the generation of plasmin may facilitate keratinocyte division , migration and differentiation . In this study we have investigated the characteristics of plasminogen binding sites in normal human epidermis . It was found that DB00513 and benzamidine displaced endogenous epidermal plasminogen from the basal layer suggesting that endogenous plasminogen binds initially via the kringle 5 aminohexyl ( AH ) site . P00747 binding sites in epidermis were further investigated by displacing endogenous plasminogen and incubating sections with exogenously added glu-plasminogen , lys-plasminogen and plasmin or the isolated plasminogen fragments kringles 1-3 , kringle 4 and kringle 5L . The results suggest that the uptake of plasminogen involves primary interaction with the kringle 5AH site and a secondary interaction with lysine binding sites of kringles 1-3 . Cell binding is not dependent upon additional reactions of the plasmin active centre . DB06813 : a novel synthetic antifolate for relapsed or refractory peripheral T-cell lymphoma and other potential uses . PURPOSE : The pharmacology , pharmacokinetics , clinical trials , adverse effects , dosage , and economic considerations of pralatrexate ( DB06813 ) are reviewed . SUMMARY : Peripheral T-cell lymphoma ( PTCL ) comprises approximately 15-20 % of all aggressive lymphomas and 5-10 % of all non-Hodgkin 's lymphomas . Advanced PTCL is often refractory to traditional first-line chemotherapy regimens . DB06813 was developed as a synthetic folate analog antimetabolite that competitively inhibits dihydrofolate reductase ( P00374 ) . This results in the depletion of thymidine , leading to interference with deoxyribonucleic acid synthesis and cancer cell death . DB06813 has a higher potency than methotrexate and edatrexate ( EDX ) . The efficacy and safety of DB06813 have been demonstrated in the PROPEL trial , a prospective phase II trial in patients with relapsed or refractory PTCL . Patients with prior stem cell transplantation receiving DB06813 also had similar response rates ( RRs ) . DB06813 was investigated on the treatment of relapsed or refractory cutaneous T-cell lymphoma , previously treated advanced non-small cell lung cancer and other solid malignancies . DB06813 has similar side effects to other P00374 inhibitors . The most common side effect of DB06813 is mucositis . The recommended dose of DB06813 is 30 mg/m(2) weekly once for 6 weeks in 7-week cycle until disease progresses or unacceptable toxicity for PTCL and may require dose reduction or discontinuation . Patients should be supplemented with oral folic acid and intramuscular vitamin B(12) injections . CONCLUSION : DB06813 provides clinical benefit to patients with relapsed or refractory PTCL with durable complete and partial responses in patients who had not responded to multiple prior treatment regimens . A review of the use of frovatriptan in the treatment of menstrually related migraine . Menstrual migraine ( MM ) is a highly prevalent condition associated with considerable disability . Migraine attacks occur exclusively around the menstrual period in approximately 10 % of women with migraine , that is , pure menstrual migraine , while at least 50 % of them also experience migraine at other times of the month , that is , menstrually related migraine ( MRM ) . The therapeutic approach to patients with MRM is based on treatment of the attack , or prophylactic strategies . Triptans are recommended as first-line treatments for moderate to severe migraine attacks , including MM . DB00998 is one of the newest triptans . Its high affinity for P28222 /1D receptors and long half-life contribute to its distinctive clinical effect , characterized by a more sustained and prolonged effect than other triptans . Indeed , frovatriptan proved to be effective in treating the acute attack , but was particularly effective in the short-term preventive therapy of MM . In addition , frovatriptan is one of the safest triptans , with the lowest risk of treatment-emergent adverse events . Following extensive evidence from randomized pharmacological trials , frovatriptan has now gained a grade A recommendation from the guidelines for short-term prophylaxis of MM . Recent post-hoc analyses of direct comparative trials also suggest that frovatriptan might have an important role in the acute treatment of MRM . In these studies , frovatriptan showed pain relief and pain-free rates similar to those of zolmitriptan , rizatriptan , and almotriptan , but with significantly lower recurrence rates . More well-designed , randomized , prospective studies , specifically enrolling women with MM , will be needed in the near future to confirm the efficacy of frovatriptan in this migraine subtype . 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 . A new clinical evidence-based gene-environment interaction model of depression . In our current understanding of mood disorders , the role of genes is diverse including the mediation of the effects of provoking and protective factors . Different or partially overlapping gene sets play a major role in the development of personality traits including also affective temperaments , in the mediation of the effects of environmental factors , and in the interaction of these elements in the development of depression . Certain genes are associated with personality traits and temperaments including e.g. , neuroticism , impulsivity , openness , rumination and extroversion . Environmental factors consist of external ( early and provoking life events , seasonal changes , social support etc. ) and internal factors ( hormones , biological rhythm generators , comorbid disorders etc ) . Some of these environmental factors , such as early life events and some prenatal events directly influence the development of personality traits and temperaments . In the NEWMOOD cohort polymorphisms of the genes of the serotonin transporter , P08908 , P28222 and 5- Q13049 and endocannabinoid P21554 receptors , tryptophan hydroxylase , P16220 , P23560 and GIRK provide evidence for the involvement of these genes in the development of depression . Based on their role in this process they could be assigned to different gene sets . The role of certain genes , such as promoter polymorphisms of the serotonin transporter ( 5-HTTLPR ) and P21554 receptor has been shown in more than one of the above factors . Furthermore , gene-gene interactions of these promoters associated with anxiety suggest the application of these polymorphisms in personalized medicine . In this review we introduce a new model including environmental factors , genes , trait and temperament markers based on human genetic studies . DB01079 for the treatment of constipation-predominant irritable bowel syndrome . DB01079 , a potent , partial serotonin 4 receptor ( Q13639 ) agonist , is an effective agent for the treatment of females with constipation-predominant irritable bowel syndrome . DB01079 enhances gastric motility , stimulates peristaltic reflux and intestinal secretion , inhibits visceral sensitivity , and/or shortens colonic transit time . This agent may help women who have failed to respond to diet and exercise , laxatives , and other forms of therapy . The optimal dose of tegaserod is 6 mg twice daily and results in decreased number of days per month with pain , bloating , and days without bowel movements . DB01079 is less effective in males than females in the treatment of constipation-predominant irritable bowel syndrome . DB01079 is well tolerated . Diarrhea is the most frequent adverse effect . The diarrhea tends to occur most frequently during the first few months of therapy and decreases with continued administration . Autocrine endocannabinoid signaling through P21554 receptors potentiates OX1 orexin receptor signaling . It has been proposed that OX(1) orexin receptors and CB(1) cannabinoid receptors can form heteromeric complexes , which affect the trafficking of OX(1) receptors and potentiate OX(1) receptor signaling to extracellular signal-regulated kinase ( P29323 ) . We have recently shown that OX(1) receptor activity releases high levels of the endocannabinoid 2-arachidonoyl glycerol ( 2-AG ) , suggesting an alternative route for OX(1)-CB(1) receptor interaction in signaling , for instance , in retrograde synaptic transmission . In the current study , we set out to investigate this possibility utilizing recombinant Chinese hamster ovary P04264 cells . 2-AG released from OX(1) receptor-expressing cells acted as a potent paracrine messenger stimulating P29323 activity in neighboring CB(1) receptor-expressing cells . When OX(1) and CB(1) receptors were expressed in the same cells , OX(1) stimulation-induced P29323 phosphorylation and activity were strongly potentiated . The potentiation but not the OX(1) response as such was fully abolished by specific inhibition of CB(1) receptors or the enzyme responsible for 2-AG generation , diacylglycerol lipase ( DAGL ) . Although the results do not exclude the previously proposed OX(1)-CB(1) heteromerization , they nevertheless unequivocally identify DAGL-dependent 2-AG generation as the pivotal determinant of the OX(1)-CB(1) synergism and thus suggest a functional rather than a molecular interaction of OX(1) and CB(1) receptors . Crystal structure of dihydrofolate reductase from Plasmodium vivax : pyrimethamine displacement linked with mutation-induced resistance . DB00205 ( Pyr ) targets dihydrofolate reductase of Plasmodium vivax ( PvDHFR ) as well as other malarial parasites , but its use as antimalarial is hampered by the widespread high resistance . Comparison of the crystal structures of PvDHFR from wild-type and the Pyr-resistant ( SP21 , DB00133 -58 --> DB00125 + DB00133 -117 --> DB00174 ) strain as complexes with NADPH and Pyr or its analog lacking p-Cl ( Pyr20 ) clearly shows that the steric conflict arising from the side chain of DB00174 -117 in the mutant enzyme , accompanied by the loss of binding to DB00133 -120 , is mainly responsible for the reduction in binding of Pyr . Pyr20 still effectively inhibits both the wild-type and SP21 proteins , and the x-ray structures of these complexes show how Pyr20 fits into both active sites without steric strain . These structural insights suggest a general approach for developing new generations of antimalarial P00374 inhibitors that , by only occupying substrate space of the active site , would retain binding affinity with the mutant enzymes . Unsaturated side chain beta-11-hydroxyhexahydrocannabinol analogs . The cannabinoid side chain is a key pharmacophore in the interaction of cannabinoids with their receptors ( P21554 and CB2 ) . To study the stereochemical requirements of the side chain , we synthesized a series of cannabinoids in which rotation around the C1'- P06681 ' bond is blocked . The key steps in the synthesis were the cuprate addition of a substituted resorcinol to (+)-apoverbenone , the TMSOTf-mediated formation of the dihydropyran ring , and the stereospecific introduction of the beta-11-hydroxymethyl group . All the analogs tested showed nanomolar affinity for the receptors , the cis-hept-1-ene side chain having the highest affinity for P21554 ( Ki = 0.89 nM ) and showing the widest separation between P21554 and CB2 affinities . The parent n-heptyl-beta-11-hydroxyhexahydrocannabinol was the least potent binding to P21554 ( Ki = 8.9 nM ) and had the lowest selectivity between P21554 and CB2 . 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 . Eupolyphaga sinensis walker displays inhibition on hepatocellular carcinoma through regulating cell growth and metastasis signaling . Tumor growth and metastasis are responsible for most cancer patients ' deaths . Here , we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis . Compared with proliferation , colony formation , transwell assay and transplantable tumor in nude mouse in vitro and vivo , eupolyphaga sinensis walker extract ( ESWE ) showed good inhibition on the SMMC-7721 cell growth and metastasis . Using genome-wide microarray analysis , we found the down-regulated growth and metastasis factors , and selected down-regulated genes were confirmed by real-time PCR . Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE . Moreover , our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ , Akt , m-TOR , Erk1/2 , MEK-2 , Raf and JNK-1 , but also increasing cyclin D1 protein levels and decreasing amount of cyclin E , cyclin B1 and cdc2 of the cycle proteins . At the same time , ESWE reduced P08253 , P14780 and P61073 , P00747 , NFκB and P04637 activities . Overall , our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC , AKT , MAPK signaling and related metastasis signaling , having potential in cancer therapy .

MH_train_1487

MH_train_1487

MH_train_1487

interacts_with DB06271?

multiple_choice

Role of antispasmodics in the treatment of irritable bowel syndrome . Irritable bowel syndrome ( IBS ) is a long-lasting , relapsing disorder characterized by abdominal pain/discomfort and altered bowel habits . Intestinal motility impairment and visceral hypersensitivity are the key factors among its multifactorial pathogenesis , both of which require effective treatment . Voltage-gated calcium channels mediate smooth muscle contraction and endocrine secretion and play important roles in neuronal transmission . Antispasmodics are a group of drugs that have been used in the treatment of IBS for decades . DB01616 citrate , a spasmolytic , decreases the sensitivity of smooth muscle contractile proteins to calcium , and it is a selective P08908 receptor antagonist . DB01616 , in combination with simethicone , has been demonstrated to effectively reduce abdominal pain and discomfort in a large placebo-controlled trial . Mebeverine is a musculotropic agent that potently blocks intestinal peristalsis . Non-placebo-controlled trials have shown positive effects of mebeverine in IBS regarding symptom control ; nevertheless , in recent placebo-controlled studies , mebeverine did not exhibit superiority over placebo . Otilonium bromide is poorly absorbed from the GI tract , where it acts locally as an L-type calcium channel blocker , an antimuscarinic and a tachykinin NK2 receptor antagonist . Otilonium has effectively reduced pain and improved defecation alterations in placebo-controlled trials in IBS patients . DB09090 bromide is also an L-type calcium channel blocker that acts locally in the GI tract . DB09090 improves motility disorders and consequently reduces stool problems in IBS patients . Phloroglucinol and trimethylphloroglucinol are non-specific antispasmodics that reduced pain in IBS patients in a placebo-controlled trial . Antispasmodics have excellent safety profiles . T-type calcium channel blockers can abolish visceral hypersensitivity in animal models , which makes them potential candidates for the development of novel therapeutic agents in the treatment of IBS . Functional induction of the cystine-glutamate exchanger system Xc(-) activity in SH-SY5Y cells by unconjugated bilirubin . We have previously reported that exposure of SH-SY5Y neuroblastoma cells to unconjugated bilirubin ( UCB ) resulted in a marked up-regulation of the mRNA encoding for the Na(+)-independent cystine∶glutamate exchanger System X(c)(-) ( Q9UPY5 and P08195 genes ) . In this study we demonstrate that SH-SY5Y cells treated with UCB showed a higher cystine uptake due to a significant and specific increase in the activity of System X(c)(-) , without the contribution of the others two cystine transporters ( X(AG)(-) and P19440 ) reported in neurons . The total intracellular glutathione content was 2 folds higher in the cells exposed to bilirubin as compared to controls , suggesting that the internalized cystine is used for gluthathione synthesis . Interestingly , these cells were significantly less sensitive to an oxidative insult induced by hydrogen peroxide . If System X(c)(-) is silenced the protection is lost . In conclusion , these results suggest that bilirubin can modulate the gluthathione levels in neuroblastoma cells through the induction of the System X(c)(-) , and this renders the cell less prone to oxidative damage . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Poly(ADP-ribose) polymerase-1 mediates angiotensin II-induced expression of plasminogen activator inhibitor-1 and fibronectin in rat mesangial cells . OBJECTIVE : To investigate the effects of poly(ADP-ribose) polymerase-1 ( P09874 ) on angiotensin II ( Ang II ) -induced plasminogen activator inhibitor-1 ( P05121 ) and fibronectin ( FN ) in rat mesangial cells ( RMCs ) . METHODS : Followed by serum starvation for 16 h , RMCs were exposed to Ang II for an indicated time to examine the protein expression of P09874 . The cells were treated with or without Ang II for 12-24 h in the presence or absence of an inhibitor of PARP , N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide hydrochloride ( PJ34 ) or small interfering RNA ( siRNA ) duplexes targeting P09874 . The mRNA and protein expressions of P09874 , P05121 and FN were determined by real-time RT-PCR and Western blot , respectively . The activity of P09874 was examined by colorimetric assay . RESULTS : Ang II did not only significantly induce P09874 expression and activity , but also increased P05121 and FN expression in RMCs . All these responses induced by Ang II were significantly inhibited by both the PARP inhibitor PJ34 and downregulating P09874 with the siRNA technique . CONCLUSIONS : Our data suggest that P09874 mediates Ang II-induced P05121 and FN in RMCs and may thus represent a potential therapeutic target in the treatment of glomerular disease . High-fat diet promotes lung fibrosis and attenuates airway eosinophilia after exposure to cockroach allergen in mice . Obesity is an important risk factor for asthma but the mechanistic basis for this association is not well understood . In the current study , the impact of obesity on lung inflammatory responses after allergen exposure was investigated . C57BL/6 mice maintained on a high-fat diet ( HFD ) or a normal diet ( ND ) after weaning were sensitized and challenged with cockroach allergen ( CRA ) . Airway inflammation was assessed based on inflammatory cell recruitment , measurement of lung Th1-Th2 cytokines , chemokines , eicosanoids , and other proinflammatory mediators as well as airway hyperresponsiveness ( P35869 ) . CRA-challenged mice fed a HFD exhibited significantly decreased allergen-induced airway eosinophilia along with reduced lung P05113 , P35225 , LTC4 , P51671 , and P13500 levels as well as reduced mucus secretion and smooth muscle mass compared to ND fed mice . However , allergen-challenged HFD fed mice demonstrated significantly increased P05121 and reduced DB00917 levels in the lung relative to corresponding ND fed mice . Interestingly , saline-exposed HFD fed mice demonstrated elevated baseline levels of TGF-β1 , arginase-1 , hypoxia-inducible factor-1α , and lung collagen expression associated with decreased lung function compared to corresponding ND fed mice . These studies indicate that a HFD inhibits airway eosinophilia while altering levels of P05121 and DB00917 in response to CRA in mice . Further , a HFD can lead to the development of lung fibrosis even in the absence of allergen exposure which could be due to innate elevated levels of specific profibrotic factors , potentially affecting lung function during asthma . 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 . Identification of antithrombin-modulating genes . Role of O95461 , a gene encoding a bifunctional glycosyltransferase , in the secretion of proteins ? The haemostatic relevance of antithrombin together with the low genetic variability of P01008 , and the high heritability of plasma levels encourage the search for modulating genes . We used a hypothesis-free approach to identify these genes , evaluating associations between plasma antithrombin and 307,984 polymorphisms in the GAIT study ( 352 individuals from 21 Spanish families ) . Despite no SNP reaching the genome wide significance threshold , we verified milder positive associations in 307 blood donors from a different cohort . This validation study suggested O95461 , a gene encoding a protein with xylosyltransferase and glucuronyltransferase activities that forms heparin-like linear polysaccharides , as a potential modulator of antithrombin based on the significant association of one SNPs , rs762057 , with anti-FXa activity , particularly after adjustment for age , sex and P01008 rs2227589 genotype , all factors influencing antithrombin levels ( p = 0.02 ) . Additional results sustained this association . O95461 silencing inHepG2 and P29320 -EBNA cells did not affect P01008 mRNA levels but significantly reduced the secretion of antithrombin with moderate intracellular retention . Milder effects were observed on α1-antitrypsin , prothrombin and transferrin . Our study suggests O95461 as the first known modifier of plasma antithrombin , and proposes a new role for O95461 in modulating extracellular secretion of certain glycoproteins . Two different P01579 nonresponsive variants derived from the B-cell lymphoma 70Z/3 . The kappa immunoglobulin ( Igk ) light chain locus is transcriptionally silent in the mouse B-cell lymphoma 70Z/3 . However , exposure to lipopolysaccharide ( LPS ) or interferon-gamma ( IFN ) causes a marked increase in Igk transcription . By immunoselection , we isolated two variants that are nonresponsive to IFN . One variant , AT7.2 , has retained its response to LPS ( IFN-LPS+ ) , whereas the other , P01008 .3 , is also nonresponsive to LPS ( IFN-LPS- ) . Stable transfection of an intact Igk gene does not rescue the phenotype of either variant . Both variants have intact Igk genes and neither is deficient in the binding or uptake of IFN . Nuclear extracts from LPS-treated wild-type 70Z/3 cells show strong increases in three transcription factors : P09086 , NF-kappa B , and kBF-A . Remarkably , when the IFN-LPS- variant is treated with LPS , all three transcription factors are still observed in the nuclear extracts . Treatment of wild-type cells with either LPS or IFN also causes a decrease in nuclear complexes that bind to two other regions of the Igk intron enhancer , the octenh and the E kappa MHCIC regions . Both of these changes are also observed after LPS or IFN treatment of the IFN-LPS- variant . Thus , this variant transduces the IFN and LPS signals at least into the nuclear compartment , but still fails to activate Igk transcription . In contrast , the IFN-LPS+ variant decreases neither the octenh nor the E kappa MHCIC binding complexes in response to IFN . This variant may be defective in transducing the IFN signal to the nucleus . These variants will be useful in studying the activation of Igk transcription and the IFN signaling pathway in B cells . 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 . 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 . A phase I study of cabozantinib ( DB05153 ) in patients with differentiated thyroid cancer . BACKGROUND : DB08875 targets tyrosine kinases including MET , vascular endothelial growth factor ( P15692 ) receptor 2 , and rearranged during transfection ( P07949 ) . Differentiated thyroid cancer ( DTC ) is a tumor type that may be sensitive to cabozantinib . Therefore , we evaluated cabozantinib in a cohort of heavily pretreated patients with metastatic DTC . METHODS : This single-arm open-label phase I trial assessed the safety , tolerability , and antitumor activity of cabozantinib in DTC patients taking part in a drug-drug interaction study . Adult patients with histologically confirmed metastatic or surgically unresectable DTC ( including papillary , follicular , or Hürthle cell ) were enrolled . Patients received daily oral dosing of 140 mg cabozantinib . Safety was assessed by evaluation of adverse events ( AEs ) , vital signs , electrocardiograms , laboratory tests , and concomitant medications . Tumor response by magnetic resonance imaging or computed tomography scan was investigator assessed using Response Evaluation Criteria In Solid Tumors ( RECIST ) v1.0 . RESULTS : The study enrolled 15 patients who had failed standard radioactive iodine therapy . Patients had received a median of two prior systemic agents , and 11 patients ( 73 % ) had previously received at least one P15692 pathway inhibiting therapy . Common AEs included diarrhea , nausea , fatigue , and decreased appetite . Partial response was reported in eight patients ( 53 % ) . Median progression-free survival and median overall survival were not reached . CONCLUSIONS : DB08875 demonstrates a safety profile similar to other multitargeted VEGFR inhibitors in advanced DTC patients . The antitumor activity observed in this study warrants further investigation of cabozantinib in patients with advanced DTC . 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 . Clinical significance of arginase after liver transplantation . Liver graft function after transplantation is dependent on ischemia-reperfusion injury , toxicity of drugs ( immunosuppression , antibiotics and other ) and transplant rejection . Although routinely monitored with enzymatic tests ( Q9NRA2 , ALT , P19440 , ALP ) , bilirubin and coagulation parameters , differentiation between these pathologies is hardly possible without liver biopsy . Arginase ( 3.5.3.1 ) mostly exists in the liver and in trace amounts in extra-hepatic tissue . Thus , we hypothesized that activity of arginase could be a more specific test of liver function . Sera of 32 liver transplant recipients were tested for Q9NRA2 , ALT , P01008 , bilirubin and arginase . Samples were obtained daily in first 2 weeks after LTx and weekly afterwards . Correlation of arginase activity with other liver function markers was calculated . Serum arginase peaked at day 1 post LTx ( mean 64,6+/-91 IU/L ) , and decreased more rapidly than other tests if good liver function was observed . The values showed strong and significant correlation with Q9NRA2 and ALT activities ( Pearsons R 0,65 and 0,47 respectively ) . We conclude that activity of arginase in the serum is an exact test of liver function . Preventing NAD(+) depletion protects neurons against excitotoxicity : bioenergetic effects of mild mitochondrial uncoupling and caloric restriction . Neurons are excitable cells that require large amounts of energy to support their survival and functions and are therefore prone to excitotoxicity , which involves energy depletion . By examining bioenergetic changes induced by glutamate , we found that the cellular nicotinamide adenine dinucleotide ( NAD(+) ) level is a critical determinant of neuronal survival . The bioenergetic effects of mitochondrial uncoupling and caloric restriction were also examined in cultured neurons and rodent brain . 2 , 4-dinitrophenol ( DNP ) is a chemical mitochondrial uncoupler that stimulates glucose uptake and oxygen consumption on cultured neurons , which accelerates oxidation of NAD(P)H to NAD(+) in mitochondria . The NAD(+)-dependent histone deacetylase sirtulin 1 ( Q96EB6 ) and glucose transporter 1 ( P11166 ) mRNA are upregulated mouse brain under caloric restriction . To examine whether NAD(+) mediates neuroprotective effects , nicotinamide , a precursor of NAD(+) and inhibitor of Q96EB6 and poly ( ADP-ribose ) polymerase 1 ( P09874 ) ( two NAD(+)-dependent enzymes ) , was employed . DB02701 attenuated excitotoxic death and preserved cellular NAD(+) levels to support Q96EB6 and PARP 1 activities . Our findings suggest that mild mitochondrial uncoupling and caloric restriction exert hormetic effects by stimulating bioenergetics in neurons thereby increasing tolerance of neurons to metabolic stress . DB06612 for severe eosinophilic asthma . In this large ( 616 patients ) , double-blind , placebo-controlled , dose-ranging study of mepolizumab ( a monoclonal antibody that blocks P05113 binding to its receptor ) , patients were given placebo , 75- , 250- or 750-mg mepolizumab by intravenous infusion every 4 weeks for 1 year . Exacerbation rates at all doses were 50 % less than those in the placebo group . There were no changes in any other asthma measures ( symptoms , quality of life or lung function ) . This may be a useful advance for a subgroup of severe asthma with frequent exacerbations and persistent eosinophilia , which may be about half of severe asthmatics . More information on patient selection and cost-benefit will be required . 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 . P25116 -mediated synovial proliferation in patients with rheumatoid arthritis . Synovial cell proliferation is one of the pathological bases of rheumatoid arthritis ( RA ) . Several cytokines including IL-1 and P05231 and growth factors have been shown to be involved in the synovial cell proliferation in RA . Thrombin is a multifunctional protease and acts as a mitogen for several cell types through its specific receptor . To assess whether thrombin is involved in overproliferation of rheumatoid synovial cells , we measured the concentration of thrombin-anti-thrombin III ( P01008 ) complex ( TAT ) in synovial fluid obtained from patients with RA or osteoarthritis ( OA ) . We also examined the effect of thrombin or thrombin receptor agonist peptide ( TRAP ) on cell growth of synovial cell clones ( SCCs ) established from an RA patient . The concentrations of TAT in the synovial fluid from patients with RA were significantly higher than in those with OA . Moreover , both thrombin and TRAP enhanced proliferation of synovial cells in vitro . We also characterized the expression of thrombin receptor mRNA by reverse transcription-PCR . The expression of mRNA for thrombin receptor was up-regulated by thrombin or TRAP stimulation . P25116 antigen was also detected on both SCCs and synovial tissue from RA patients by immunostaining using a monoclonal antibody against thrombin receptor . These findings indicate that thrombin may act as a mitogen for synovial cells through thrombin receptor and may play some role in synovial overproliferation and remodeling in RA . 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 . 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 . 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 . 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 .

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interacts_with DB09073?

multiple_choice

The effects of GM- P04141 , steel factor and MIP-1alpha on the expression and activation of Cdc25A phosphatase in Mo7e cells . Active cyclin-dependent kinases ( CDKs ) are required for progression through the P55008 phase of the cell cycle and entry into S phase . Activity of P55008 CDKs is controlled by mechanisms including phosphorylation of Thr14 and Tyr15 residues . Removal of inhibitory phosphates on these amino acid residues is required for P55008 CDK activation , and is mediated by the Cdc25A phosphatase . Regulation of active Cdc25A phosphatase levels may be important for the proliferation of hematopoietic progenitor cells , effects assessed in the human growth-factor-dependent cell line Mo7e . Constitutive Cdc25A protein levels were enhanced with granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) plus steel factor ( SF ) . Cdc25A is thought to exert its activity in the nucleus , and nuclear protein levels of Cdc25A were also enhanced with GM- P04141 and SF . GM- P04141 plus SF promote synergistic growth of Mo7e cells . Pretreatment with macrophage inflammatory protein ( MIP-1alpha ) inhibited GM- P04141 - plus SF-induced growth and upregulation of Cdc25A protein levels . Stimulation with GM- P04141 and SF also rapidly increased Cdc25A phosphatase activity , an effect suppressed by MIP-1alpha . A concomitant inhibition of increased P11802 kinase activity correlated with increased phosphotyrosine levels on P11802 when cells were pretreated with MIP-1alpha prior to GM- P04141 and SF . These data suggest that Cdc25A expression and activity are regulated during proliferation of Mo7e cells . Korean Red DB01404 Saponin Fraction Downregulates Proinflammatory Mediators in LPS Stimulated RAW264.7 Cells and Protects Mice against Endotoxic Shock . Korean red ginseng has shown therapeutic effects for a number of disease conditions . However , little is known about the antiinflammatory effect of Korean red ginseng saponin fraction ( RGSF ) in vitro and in vivo . Therefore , in this study , we showed that RGSF containing 20(S)-protopanaxadiol type saponins inhibited nitric oxide production and attenuated the release of tumor necrotic factor ( P01375 ) -α , interleukin ( IL ) -6 , granulocyte monocyte colony stimulating factor ( P04141 ) , and macrophage chemo-attractant protein-1 in lipopolysaccharide ( LPS ) stimulated murine macrophage RAW264.7 cells . Moreover , RGSF down-regulated the mRNA expressions of inducible nitric oxide synthase , cyclooxyginase-2 , IL-1β , P01375 -α , P04141 , and P05231 . Furthermore , RGSF reduced the level of P01375 -α in the serum and protected mice against LPS mediated endotoxic shock . In conclusion , these results indicated that ginsenosides from RGSF and their metabolites could be potential sources of therapeutic agents against inflammation . 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 . P10646 in activated prothrombin complex concentrates ( aPCC ) moderates the effectiveness of therapy in some severe hemophilia A patients with inhibitor . Some hemophilia A patients who have developed inhibitors are poorly responsive to activated prothrombin complex concentrates ( aPCC ) after daily dosage , but the mechanism(s) underlying this remain unknown . We examined two representative cases . In case 1 , we found that changing to recombinant factor VIIa ( DB00036 ) therapy was more effective , and the response to aPCC was restored within ~2 weeks . P13726 ( TF ) -triggered thrombin generation demonstrated a prolonged lag-time and decreased peak thrombin , and this impairment was focused on TF pathway inhibitor ( P10646 ) . Plasma-free P10646 was elevated post-infusion of aPCC , while this was unaffected by DB00036 . P10646 returned to normal range within 2-3 weeks . Plasmas obtained from patients with poor or good response to aPCC ( aPCC-poor or aPCC-good ) , and good response to DB00036 ( FVIIa-good ) demonstrated that free P10646 levels are increased in both aPCC groups , but not in FVIIa-good . P10646 levels pre- and post-infusion in aPCC-poor were significantly higher than those in aPCC-good . Addition of anti- P10646 antibody to the reaction samples demonstrated a greater increase of peak thrombin in aPCC-poor compared to aPCC-good , showing the higher P10646 activity in aPCC-poor . Free P10646 contained in aPCC corresponded to the increasing levels in plasma . In conclusion , P10646 in aPCC attenuated thrombin generation , and the reduced effectiveness of therapy in these circumstances appeared to be related to P10646 activity . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . DB01088 has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 + T cells was measured by ELISA . The expression of the transcription factor FoxP3 after co-culture of DCs with P01730 + CD45RA+ T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 , P05231 , P10145 and IL-12p70 in mDCs , while it enhanced P22301 production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA+ T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP3 expression and P22301 production . Additionally , iloprost inhibited the MIP-3beta-induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans . DB06096 , a selective P29475 inhibitor and a P28222 /1D receptor agonist , inhibits P80511 release in preclinical migraine models . BACKGROUND : DB06096 is a combined neuronal nitric oxide synthase ( P29475 ) inhibitor and 5-hydroxytryptamine 1B/1D ( P28222 /1D ) receptor agonist . Using preclinical models , we evaluated whether these two unique therapeutic principles have a synergistic effect in attenuating stimulated calcitonin gene-related peptide ( P80511 ) release , a marker of trigeminal activation . METHODS : We examined the effect of DB06096 on : ( 1 ) DB00761 - , capsaicin- and resiniferatoxin ( RTX ) -induced immunoreactive P80511 ( iCGRP ) release from isolated preparation of rat dura mater , trigeminal ganglion ( TG ) and trigeminal nucleus caudalis ( P24821 ) ; and ( 2 ) capsaicin- and electrical stimulation ( ES ) -induced middle meningeal artery ( MMA ) dilation in a rat closed-cranial window . RESULTS : DB06096 inhibited : ( 1 ) DB00761 -stimulated iCGRP release from dura mater ( % decrease mean ± SEM , lowest effective concentration ) ( 35 ± 6 % , 30 µM ) , TG ( 24 ± 11 % , 10 µM ) and P24821 ( 40 ± 8 % , 10 µM ) ; ( 2 ) capsaicin- and RTX-induced iCGRP release from dura mater ; and ( 3 ) capsaicin- and ES-induced increase in dural artery diameter ( 32 ± 5 % , 3 mg kg(-1) intravenous ( i.v. ) and 36 ± 1 % , 10 mg kg(-1) i.v. ) . CONCLUSIONS : DB06096 inhibits P80511 release from migraine-relevant cephalic tissues . Its effect is most likely mediated via a combination of P29475 -inhibition and P28222 /1D receptor agonism in dura mater while the mechanisms of action for inhibition of P80511 release from TG and P24821 have to be investigated further . JTE-522 , a selective P35354 inhibitor , inhibits cell proliferation and induces apoptosis in RL95-2 cells . AIM : To investigate whether JTE-522 [ 4-(4-cyclohexyl-2-methyloxazol-5-yl)-2-fluorobenzenesulfonamide ] , a selective P35354 inhibitor , can induce apoptosis and inhibit cell proliferation in human endometrial cancer cell line RL95-2 cells and to explore the molecular mechanisms . METHODS : [ 3-(4,5)-dimethylthiazol-2-yl ] -2,5-diphenyl tetrazolium bromide ( MTT ) , DNA ladder , enzyme-linked immunosorbent assay ( ELISA ) , flow cytometry , RT-PCR , and Western blot analysis were employed to investigate effect of JTE-522 on human endometrial cancer cell line RL95-2 cells and the related molecular mechanisms . RESULTS : JTE-522 inhibited the growth of RL95-2 cells and induced the apoptosis . Furthermore , it arrested G0/ P55008 phase and inhibited S phase in RL95-2 cells . JTE-522 inhibited the expressions of P35354 mRNA , phosphorylated Rb , and P11802 proteins , while increased the levels of p53 , P38936 , cyclin D1 proteins , and the activity of caspase-3 in RL95-2 cells . CONCLUSION : JTE-522 inhibits cell proliferation and induces apoptosis in RL95-2 cells , which may be associated with the activation of caspase-3-like proteases , down-regulation of the expression of P35354 mRNA , phosphorylated Rb , and P11802 proteins , and up-regulation of the expressions of p53 , P38936 , and cyclin D1 proteins . The effects of nepafenac and amfenac on retinal angiogenesis . PURPOSE : DB06802 is a potent NSAID that rapidly penetrates the eye following topical ocular administration . In the eye , nepafenac is converted to amfenac , which has unique time-dependent inhibitory properties for P23219 and P35354 . The purpose of the present study was to investigate the capacity of amfenac to inhibit discrete aspects of the angiogenic cascade in vitro , and to test the efficacy of amfenac and nepafenac in vivo , using the rat OIR model . METHODS : Müller cells were treated with amfenac , celecoxib ( P35354 ) , or SC-560 ( P23219 ) , and hypoxia-induced P15692 and PGE(2) assessed . Endothelial cells were treated with amfenac , celecoxib , or SC-560 , and P15692 -induced proliferation and tube formation assessed . Rat pups were subjected to OIR , received intravitreal injections of amfenac , celecoxib , or SC-560 , and neovascularization ( NV ) , prostanoid production , and P15692 assessed . Other OIR-exposed pups were treated with topical nepafenac , ketorolac , or diclofenac , and inhibition of NV assessed . RESULTS : Amfenac treatment failed to inhibit hypoxia-induced P15692 production . Amfenac treatment significantly inhibited P15692 -induced tube formation and proliferation by EC . Amfenac treatment significantly reduced retinal prostanoid production and NV in OIR . DB06802 treatment significantly reduced retinal NV in OIR ; ketorolac and diclofenac had no effect . CONCLUSIONS : DB06802 and amfenac inhibit OIR more effectively than the commercially available topical and injectable NSAIDs used in this study . Our data suggests there are P36551 -dependent and P36551 -independent mechanisms by which amfenac inhibits OIR . Because it is bioavailable to the posterior segment following topical delivery , nepafenac appears to be a promising advancement in the development of therapies for neovascular eye diseases . Secreted cyclophilin A , a peptidylprolyl cis-trans isomerase , mediates matrix assembly of hensin , a protein implicated in epithelial differentiation . Q9UGM3 is a rabbit ortholog of Q9UGM3 , a multifunctional , multidomain protein implicated in the regulation of epithelial differentiation , innate immunity , and tumorigenesis . Q9UGM3 in the extracellular matrix ( Q13201 ) induced morphological changes characteristic of terminal differentiation in a clonal cell line ( clone C ) of rabbit kidney intercalated cells . Although hensin is secreted in monomeric and various oligomeric forms , only the polymerized Q13201 form is able to induce these phenotypic changes . Here we report that hensin secretion and matrix assembly were inhibited by the peptidylprolyl cis-trans isomerase ( PPIase ) inhibitors cyclosporin A ( DB00091 ) and a derivative of cyclosporin A with modifications in the d- DB00133 side chain ( Cs9 ) but not by the calcineurin pathway inhibitor FK506 . PPIase inhibition led to failure of hensin polymerization in the medium and Q13201 , plus the loss of apical cytoskeleton , apical microvilli , and the columnar epithelial shape of clone C cells . P62937 was produced and secreted into the media to a much greater extent than cyclophilins B and C . Our results also identified the direct DB00091 -sensitive interaction of cyclophilin A with hensin , suggesting that cyclophilin A is the PPIase that mediates the polymerization and matrix assembly of hensin . These results are significant because this is the first time a direct role of peptidylprolyl cis-trans isomerase activity has been implicated in the process of epithelial differentiation . 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 . Q8NFT8 protein antagonist JQ1 is synergistically lethal with P36888 tyrosine kinase inhibitor ( TKI ) and overcomes resistance to P36888 -TKI in AML cells expressing P17948 -ITD . Recently , treatment with bromodomain and extraterminal protein antagonist ( BA ) such as JQ1 has been shown to inhibit growth and induce apoptosis of human acute myelogenous leukemia ( AML ) cells , including those expressing P36888 -ITD . Here , we demonstrate that cotreatment with JQ1 and the P36888 tyrosine kinase inhibitor ( TKI ) ponatinib or DB05213 synergistically induce apoptosis of cultured and primary P28906 (+) human AML blast progenitor cells ( BPC ) expressing P36888 -ITD . Concomitantly , as compared with each agent alone , cotreatment with JQ1 and the P36888 -TKI caused greater attenuation of c-MYC , P10415 , and P11802 /6 . Simultaneously , cotreatment with JQ1 and the P36888 -TKI increased the levels of P38936 , O43521 , and cleaved PARP , as well as mediated marked attenuation of p- P42229 , p-AKT , and p- P27361 /2 levels in AML BPCs . Conversely , cotreatment with JQ1 and P36888 -TKI was significantly less active against P28906 (+) normal bone marrow progenitor cells . Knockdown of O60885 by short hairpin RNA also sensitized AML cells to P36888 -TKI . JQ1 treatment induced apoptosis of mouse Ba/ P13726 cells ectopically expressing P36888 -ITD with or without P36888 -TKI-resistant mutations F691L and D835V . Compared with the parental human AML P36888 -ITD-expressing MOLM13 , MOLM13-TKIR cells resistant to DB05213 were markedly more sensitive to JQ1-induced apoptosis . Furthermore , cotreatment with JQ1 and the pan-histone deacetylase inhibitor ( HDI ) DB06603 synergistically induced apoptosis of P36888 -TKI-resistant MOLM13-TKIR and MV4-11-TKIR cells . Collectively , these findings support the rationale for determining the in vivo activity of combined therapy with BA and P36888 -TKI against human AML cells expressing P36888 -ITD or with BA and HDI against AML cells resistant to P36888 -TKI . A combination of molecular cytogenetic analyses reveals complex genetic alterations in conventional renal cell carcinoma . Here we report the complex pattern of genomic imbalances and rearrangements in a panel of 19 renal cell carcinoma cell lines detected with molecular cytogenetic analysis . Consistent heterogeneity in chromosome number was found , and most cell lines showed a near-triploid chromosome complement . Several cell lines showed deletions of the P04637 ( alias p53 ) , CDKN2A ( alias p16 ) , and P40337 genes . Multiplex fluorescence in situ hybridization ( M- Q5TCZ1 ) analysis revealed chromosome 3 translocated to several other partners chromosomes , as well as breakage events commonly affecting chromosomes 1 , 5 , 8 , 10 , and 17 . The most common abnormality detected with comparative genomic hybridization ( CGH ) was deletions of chromosome 3p , with loss of the Q9NS23 , P49789 , and p44S10 loci frequently involved . CGH gain of 5q showed overrepresentation of the P18146 and P07333 genes . Recurrent alterations to chromosome 7 included rearrangement of 7q11 and gains of the P00533 , O15164 , and P35250 genes . Several lines exhibited rearrangement of 12q11 approximately q14 and overrepresentation of P11802 and SAS loci . M- Q5TCZ1 revealed several other recurrent translocations , and CGH findings included loss of 9p , 14q , and 18q and gain of 8q , 12 , and 20 . Further genomic microarray changes included loss of Q13126 , IGH@ , P28222 , and Q13485 ( previously Q13485 ) and gains of MYC and P11387 . An excellent correlation was observed between the genomic array and Q5TCZ1 data , demonstrating that this technique is effective and accurate . The aberrations detected here may reflect important pathways in renal cancer pathogenesis . 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 . [ Detection of a high-affinity prostaglandin I2 binding site in the human thyroid ] . This study is concerned with the identification and the pharmacological properties of P43119 binding sites on human thyroid membrane fractions . Scatchard analysis is not linear , revealing a high- and a low-affinity receptor binding site . ( 3 H ) DB01088 binding experiments were performed under various clinical conditions : in thyroid cancer the low-affinity binding sites disappear totally and the specific high-affinity binding sites are diminished according to the grade of differentiation of the cancer . An alteration in Bmax and Kd is also observed in cold nodules , in Hashimoto 's and Riedl 's thyroiditis and in hyperthyroidism , whereas hot nodules exhibit an increase in both the receptor subpopulations . The data provide evidence for specific DB01240 binding sites and support the suggestion of a direct regulatory key-role of DB01240 in thyroid intermediary metabolism . DB09073 ( PD 0332991 ) : targeting the cell cycle machinery in breast cancer . INTRODUCTION : The cyclin D-cyclin-dependent kinases 4 and 6 ( P11802 /6 ) -retinoblastoma ( P06400 ) pathway , governing the cell cycle restriction point , is frequently altered in breast cancer and is a potentially relevant target for anticancer therapy . DB09073 ( PD 0332991 ) , a potent and selective inhibitor of P11802 and Q00534 , inhibits proliferation of several P06400 -positive cancer cell lines and xenograft models . AREAS COVERED : The basic features and abnormalities of the cell cycle in breast cancer are described , along with their involvement in estrogen signaling and endocrine resistance . The pharmacological features of palbociclib , its activity in preclinical models of breast cancer and the potential determinants of response are then illustrated , and its clinical development in breast cancer described . A literature search on the topic was conducted through PubMed and the proceedings of the main cancer congresses of recent years . EXPERT OPINION : The combination of palbociclib with endocrine agents is a very promising treatment and Phase III clinical trials are ongoing to confirm its efficacy . Further , potentially useful combinations are those with drugs targeting mitogenic signaling pathways , such as P04626 - and PI3K-inhibitors . Combination with chemotherapy seems more problematic , as antagonism has been reported in preclinical models . The identification of predictive factors , already explored in preclinical studies , must be further refined and validated in clinical trials . Human metabotropic glutamate receptor 2 gene ( Q14416 ) : chromosomal sublocalization ( 3p21.1- P38936 .2 ) and genomic organization . Imbalances in glutamatergic function have been implicated in the pathogenesis of neuropsychiatric disorders . Consequently , glutamate receptors genes are promising candidates in search of susceptibility genes for these disorders . In the present study , we report the chromosomal sublocalization and genomic organization of the human metabotropic glutamate receptor 2 gene ( Q14416 ) . Using monochromosomal hybrid cell lines of NIGMS Mapping Panel 2 ( Coriell Cell Repository ) , the Q14416 gene was localized to human chromosome 3 , confirming previously reported localization . In addition , using the radiation hybrid panel RH3 ( Research Genetics ) , we sublocalized the Q14416 gene to chromosomal region 3p21.1- P38936 .2 . The genomic organization of the Q14416 gene was established using a premade library of adaptor-ligated , human-specific genomic DNA fragments . The gene consists of 5 exons , with sizes ranging from 74 to 1,076 bp . Mechanism of the induction of the differentiation of HL-60 leukemia cells by antifolates . The classic inhibitor of dihydrofolate reductase ( P00374 ) , methotrexate ( MTX ) , has been shown to be an effective inducer of the differentiation of HL-60 promyelocytic leukemia cells ( Bodner A.J. et al. ; J. Natl. Cancer Inst. 67:1025-1030 ; 1981 ) . We have obtained evidence that induction of the differentiation of these cells by MTX , as well as by other folic acid antagonists , is the result of the effects of these agents on purine and thymine nucleotide biosynthesis . DB04485 ( 10 microM ) completely blocked both the cytotoxicity and induction of differentiation produced by the specific inhibitor of thymidylate synthase ( TS ) , N10-propargyl-5,8-dideazafolic acid ( CB-3717 ) . DB04485 also blocked the acute cytotoxicity caused by MTX and trimetrexate ( DB01157 ) ; the induction of differentiation and the loss of proliferative capacity , however , were only partially prevented by thymidine . DB04076 ( 100 microM ) , which completely restored antifolate-depleted purine nucleotide levels , had no effect on either the cytotoxicity or the induction of maturation produced by these agents . The growth inhibitory effects and the induction of differentiation caused by dideazatetrahydrofolic acid ( DDATHF ) , which acts on de novo purine nucleotide biosynthesis rather than on P00374 or TS , was completely prevented by hypoxanthine . DB04076 also completely prevented the inhibition of cellular replication and induction of differentiation by MTX and DB01157 when combined with thymidine . The findings suggest that the depletion of intracellular thymine nucleotide levels by the antifolates , MTX , DB01157 , and CB-3717 is the primary event involved in the maturation of HL-60 leukemia cells produced by these agents and that maturation occurs concomitantly with a high level of cytotoxicity. ( ABSTRACT TRUNCATED AT 250 WORDS ) 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 . Further evidence for a functional role of the glutamate receptor gene Q14832 in schizophrenia . In recent years , evidence has been accumulating indicating a major role of glutamate in the pathogenesis and pathophysiology of schizophrenia . Of particular importance in this regard are the metabotropic glutamate receptors ( GRM ) . Thus , a recently published trial of the amino acid analogue DB05096 , which exerts its effects through the activation of the glutamate receptors Q14832 / Q14416 , showed an improvement of positive and negative symptoms comparable to treatment with olanzapine . A functional variant of Q14832 has been described which modulates synaptic glutamate levels . We assessed whether this functional variant rs6465084 is related to schizophrenia in a large sample of patients and controls . We found an increased frequency of the A allele ( p=0.027 ) and the AA genotype ( p=0.024 ) in schizophrenia patients . Moreover , in an assessment of schizophrenia endophenotypes , patients of the AA genotype performed poorly in the digit symbol test , a measure of attention ( p=0.008 ) . Our results provide further evidence for the potential importance of the glutamate receptor Q14832 in schizophrenia , and indicate that the novel antipsychotic DB05096 may actually be targeting a pathogenic pathway of schizophrenia . 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 .

MH_train_1489

MH_train_1489

MH_train_1489

interacts_with DB00734?

multiple_choice

Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Increased cytotoxicity of an unusual DNA topoisomerase II inhibitor compound C-1305 toward HeLa cells with downregulated P09874 activity results from re-activation of the p53 pathway and modulation of mitotic checkpoints . Our previous studies have shown that murine fibroblast cells , in which P09874 gene was inactivated by gene disruption , are extremely sensitive to triazoloacridone compound C-1305 , an inhibitor of DNA topoisomerase II with unusual properties . Here , we show that pharmacological inhibition of P09874 activity by its inhibitor compound DB02690 , sensitizes human cervical carcinoma HeLa cells to compound C-1305 compared to treatment with drug alone . Cytotoxic effect of drug/ DB02690 of other topoisomerase II inhibitors varied depending on the dose of P09874 inhibitor . Increased cytotoxicity of topoisomerase II inhibitor/ DB02690 combinations was attributable to the re-activation of the p53 pathway in drug-treated HeLa cells . This lead to a more stringent cell cycle checkpoint control during G2 and M and enhanced cell death by mitotic catastrophe induced by drug/ DB02690 combinations . Interestingly , treatment of HeLa cells with DB02690 alone also increased p53 expression . This effect is , at least in part , related to the inhibition of proteasome activity by drug treatments . Together , our results show that concomitant inhibition of topoisomerase II and P09874 leads to the synergistic cytotoxic effect toward tumor cells that may be important for combination therapies with DB02690 and topoisomerase II inhibitors . We also confirmed our earlier work and show the important role of P09874 activity in the maintenance of the G2 arrest induced by DNA damaging drugs . Finally , based on our studies we propose that DB02690 and possibly other inhibitors of P09874 may be used as non-genotoxic agents to activate p53 in tumor cells with non-functional p53 pathways . 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 . Pharmacogenetics of asthma . PURPOSE OF REVIEW : Patient response to the asthma drug classes , bronchodilators , inhaled corticosteroids and leukotriene modifiers , are characterized by a large degree of heterogeneity , which is attributable in part to genetic variation . Herein , we review and update the pharmacogenetics and pharmaogenomics of common asthma drugs . RECENT FINDINGS : Early studies suggest that bronchodilator reversibility and asthma worsening in patients on continuous short-acting and long-acting beta-agonists are related to the Gly16Arg genotype for the P07550 . More recent studies including genome-wide association studies implicate variants in other genes contribute to bronchodilator response heterogeneity and fail to replicate asthma worsening associated with continuous beta-agonist use . Genetic determinants of the safety of long-acting beta-agonist require further study . Variants in P34998 , Q9UL17 , and P06734 contribute to variability in response for lung function , airways responsiveness , and exacerbations in patients taking inhaled corticosteroids . Variants in P09917 , P09960 , Q16873 , P33527 , Q9NS75 , and O94956 contribute to variability in response to leukotriene modifiers . SUMMARY : Identification of novel variants that contribute to response heterogeneity supports future studies of single nucleotide polymorphism discovery and include gene expression and genome-wide association studies . Statistical models that predict the genomics of response to asthma drugs will complement single nucleotide polymorphism discovery in moving toward personalized medicine . 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 . 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 . 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 . 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 . 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 . Non-alcoholic fatty liver induces insulin resistance and metabolic disorders with development of brain damage and dysfunction . In the present study we investigated the effect of the non-alcoholic fatty liver disease ( NAFLD ) on the alterations in the activity of neurotransmitters catabolizing enzymes and energy catabolising enzymes , prooxidants , endogenous antioxidants and proinflammatory cytokines in brain tissue of NAFLD rats . Rats were intraperitonealy injected with CCl4 solution at a dose of ( 0.021 mole/Kg , 20 μL , body weight ) three times weekly for four weeks . DB03128 esterase ( P22303 ) , monoamine oxidase ( MAO ) , prooxidant/ antioxidants status , ATPase , lipid profile and glucose level were estimated spectrophotometrically while inflammatory markers ; interleukin 6 and tumor necrosis factor alpha ( P05231 and P01375 -α ) and insulin were assessed by ELISA technique . Our results showed that the induced NAFLD and insulin resistance ( IR ) were accompanied with hyperglycemia and hyperlipidemia and lowered brain glucose level with elevated ATPase activity , prooxidant status ( TBARS level , xanthine oxidase and cytochrome 2E1 activities ) , and inflammatory markers . Through the induction period P22303 activity was significantly increased compared to control in blood , liver and brain tissues . Also , MAO activity was significantly increased in both brain and liver tissue but decreased in serum compared with control . These biochemical data were supported with pathophysiological analysis that showed severe neurodegeneration , pyknosis acuolations and cavitations . These observations warrant the reassessment of the conventional concept that the NAFLD with IR progression may induce disturbances in activities of neurotransmitters catabolising enzymes and energy production accompanied with oxidative stress and metabolic disorders , acting as relative risk factors for brain dysfunction and damage with the development of age-associated neurodegenerative diseases such as Alzheimer 's disease . 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 . 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 . Alpha-lipoic acid attenuates insulin resistance and improves glucose metabolism in high fat diet-fed mice . AIM : To investigate whether alpha-lipoic acid ( ALA ) could attenuate the insulin resistance and metabolic disorders in high fat diet-fed mice . METHODS : Male mice were fed a high fat diet ( HFD ) plus ALA ( 100 and 200 mg·kg(-1)·d(-1) ) or HFD plus a positive control drug metformin ( 300 mg·kg(-1)·d(-1) ) for 24 weeks . During the treatments , the relevant physiological and metabolic parameters of the mice were measured . After the mice were euthanized , blood samples and livers were collected . The expression of proteins and genes related to glucose metabolism in livers were analyzed by immunoblotting and real time-PCR . RESULTS : HFD induced non-alcoholic fatty liver disease ( NAFLD ) and abnormal physiological and metabolic parameters in the mice , which were dose-dependently attenuated by ALA . ALA also significantly reduced HFD-induced hyperglycemia and insulin resistance in HFD-fed mice . Furthermore , ALA significantly upregulated the glycolytic enzymes GCK , HK-1 and PK , and the glycogen synthesis enzyme GS , and downregulated the gluconeogenic enzymes PEPCK and P35575 , thus decreased glucose production , and promoted glycogen synthesis and glucose utilization in livers . Moreover , ALA markedly increased P31749 /Akt and GSK3β phosphorylation , and nuclear carbohydrate response element binding protein ( Q9NP71 ) expression in livers . Metformin produced similar effects as ALA in HFD-fed mice . CONCLUSION : ALA is able to sustain glucose homeostasis and prevent the development of NAFLD in HFD-fed mice . 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 . 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 . 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 . Leukotriene C4 production during hypoxic pulmonary vasoconstriction in isolated rat lungs . Leukotriene inhibitors preferentially inhibit hypoxic pulmonary vasoconstriction in isolated rat lungs . If lipoxygenase products are involved in the hypoxic pressor response they might be produced during acute alveolar hypoxia and a leukotriene inhibitor should block both the vasoconstriction and leukotriene production that occurs in response to hypoxia . We investigated in isolated blood perfused rat lungs whether leukotriene C4 ( LTC4 ) could be recovered from whole lung lavage fluid during ongoing hypoxic vasoconstriction . Lung lavage from individual rats had slow reacting substance ( SRS ) -like myotropic activity by guinea pig ileum bioassay . Pooled lavage ( 10 lungs ) as analyzed by reverse phase high performance liquid chromatography had an ultraviolet absorbing component at the retention time for LTC4 . At radioimmunoassay , and SRS myotropic activity by bioassay . LTC4 was not found during normoxic ventilation , during normoxic ventilation after a hypoxic pressor response , or during vasoconstriction elicited by DB00761 . DB00711 citrate , a leukotriene synthesis blocker , concomitantly inhibited the hypoxic vasoconstriction and LTC4 production . Thus P09917 products may play a role in the sequence of events leading to hypoxic pulmonary vasoconstriction . Protein indicators for HaCaT cell damage induced by UVB irradiation . UVB ( 290-320 nm ) is one major risk factor of skin diseases in human . In order to provide potential protein molecules in skin cell damage and skin diseases induced by UVB irradiation , the differentially expressed proteins in human keratinocytes cell HaCaT by UVB irradiation were screened by two-dimensional difference in-gel electrophoresis ( 2D DIGE ) combined to high performance liquid chromatography-nano-electrospray ionization tandem mass spectrometry ( HPLC-nESI-MS/MS ) . 31 protein spots were found differentially expressed with statistical significance ( p < 0.05 ) . Sixteen and 15 protein spots were observed up-regulated and down-regulated in UVB-irradiated HaCaT , respectively . Twenty-eight unique proteins were identified by searching the MS/MS data against NCBInr database through TurboSequest Bioworks software . Among the identified 28 UVB irradiation responding protein indicators , only laminin receptor 1 ( P08865 ) , calmodulin ( P62158 ) , cathepsin D ( P07339 ) and peroxiredoxin ( Q06830 ) proteins have been reported associated with skin burn and wound healing , keratinocytes proliferation and migration and epidermal barrier repairing . Most of these targets were for the first time revealed to be associated with skin cell damage induced by UVB irradiation . Function and bioinformatics analyses of the identified protein candidates were also performed using PANTHER analysis with the aid of DAVID platform . The current work provides potential protein indicators for skin cell damage from UVB irradiation . Long-term potentiation in the nucleus accumbens requires both Q12879 - and Q13224 -containing N-methyl-D-aspartate receptors . N-methyl-d-aspartate ( DB01221 ) receptors play crucial roles in several forms of long-term changes in the efficacy of glutamatergic synaptic transmission . The suggestion that the Q12879 subunit of the DB01221 receptor may be selectively involved in the induction of long-term potentiation ( LTP ) in the hippocampus and cortex has been challenged . However , the contribution of Q13224 in the induction of LTP is not always clearly established . The present study investigates the role of Q12879 and Q13224 in the induction of LTP in the nucleus accumbens ( NAc ) , a brain region that expresses high levels of Q13224 and an DB01221 -dependent form of LTP . We recorded extracellular field excitatory postsynaptic potentials/population spikes in slices of mouse NAc . High-frequency stimulation of glutamatergic fibers consistently induced LTP of the field excitatory postsynaptic potential/population spike in the NAc . LTP was abolished in the presence of selective antagonists of either Q13224 [ R-(R*,S*)-alpha-(4-hydroxyphenyl)-beta-methyl-4-(phenyl-methyl)-1-piperidine propanol and DB08954 ] or Q12879 ( [ ( R ) - [ ( S ) -1-(4-bromo-phenyl)-ethylamino ] -(2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-5-yl)-methyl ] -phosphonic acid ) subunits . Recordings performed in a low concentration of Mg(2+) ions in the perfusion solution did not reveal a selective involvement of a particular DB01221 receptor subunit because either Q12879 or Q13224 antagonists were able to block LTP . LTP was also abolished in the presence of a low concentration of the non-subunit-selective DB01221 receptor antagonist dl-2-amino-5-phosphonopentanoic acid in normal Mg(2+) and low Mg(2+) in the perfusion solution . These results show that the degree of DB01221 receptor activation , and not their subunit composition , determines whether LTP is induced in the NAc . 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 . 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 . 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_1490

MH_train_1490

MH_train_1490

interacts_with DB08879?

multiple_choice

Suppression of nuclear ADP-ribosyltransferase activity in regenerating rat liver by 5-azacytidine and its relevance to the nuclear methylating activities . A single administration of 5-azacytidine ( 5- P10323 ) to partially hepatectomized rats 24 h following operation resulted in a dose-dependent reduction of nuclear ADP-ribosyltransferase ( P09874 ) activity in the liver , when assayed after the nuclei were isolated 22 h after injection . No such a suppression by 5- P10323 was observed in the liver of intact rats . DB02097 , a known agent which prevents the incorporation of 5- P10323 into DNA , abolished the suppression of P09874 , when it was given in combination with 5- P10323 . The 5- P10323 suppressed nuclei from regenerating liver showed no decreased DNA methylating activity , as estimated from the rate of radiolabel transfer from [methyl-3H] DB00118 to the bulk DNA . The methylation of nuclear RNA and protein was markedly reduced . These results suggest that the incorporation of 5- P10323 into nucleic acids inactivates chromatin-bound P09874 without inhibition of DNA methylation . A double-blind , placebo-controlled outpatient trial of pergolide for cocaine dependence . Results of preclinical studies suggest that pergolide , a mixed D(1)/ P14416 agonist , may be useful in treating cocaine dependence . To empirically investigate this possibility , we conducted a 5-year , double-blind , placebo-controlled clinical trial of two doses of pergolide ( 0.05 and 0.25 mg bid ) in subjects with cocaine dependence and combined cocaine/alcohol dependence . Data analysis was performed on an intent to treat population ( N=358 ) and a per protocol population ( N=108 ) with urine drug screens ( UDS ) used as the main outcome measure . There were no significant effects on UDS at either pergolide dose . DB01186 had no significant effect on alcohol use in the comorbid alcohol/cocaine dependence group . DB01186 does not appear to have clinical value in the treatment of cocaine dependence or in decreasing alcohol use in cocaine-dependent individuals at the presently studied doses . Gene transcription abnormalities in canine atopic dermatitis and related human eosinophilic allergic diseases . Canine atopic dermatitis ( AD ) is clinically similar to human AD , implicating it as a useful model of human eosinophilic allergic disease . To identify cutaneous gene transcription changes in relatively early inflammation of canine AD , microarrays were used to monitor transcription in normal skin ( n=13 ) and in acute lesional AD ( P13716 ) and nearby visibly nonlesional AD ( NLAD ) skin ( n=13 ) from dogs . Scanning the putative abnormally transcribed genes , several potentially relevant genes , some abnormally transcribed in both NLAD and P13716 ( e.g. P05231 , Q8NET5 , Q9UJ68 , and P43405 ) , were observed . Comparison for abnormally transcribed genes common to two related human diseases , human AD and asthmatic chronic rhinosinusitis with nasal polyps ( aCRSwNP ) , further identified genes or gene sets likely relevant to eosinophilic allergic inflammation . These included : ( 1 ) genes associated with alternatively activated monocyte-derived cells , including members of the monocyte chemotactic protein ( MCP ) gene cluster , ( 2 ) members of the IL1 family gene cluster , ( 3 ) eosinophil-associated seven transmembrane receptor Q14246 and Q9BY15 genes , ( 4 ) interferon-inducible genes , and ( 5 ) keratin genes associated with hair and nail formation . Overall , numerous abnormally transcribed genes were observed only in canine AD ; however , many others are common to related human eosinophilic allergic diseases and represent therapeutic targets testable in dogs with AD . The kinase inhibitor staurosporine induces P55008 arrest at two points : effect on retinoblastoma protein phosphorylation and cyclin-dependent kinase 2 in normal and transformed cells . DB02010 ( ST ) , a protein kinase inhibitor , at a concentration of 20 nM arrests normal diploid fibroblasts 3 h into P55008 ( H. A. Crissman et al. , Proc. Natl. Acad. Sci. USA , 88 : 7580-7584 , 1991 ; K. Abe et al. , Exp. Cell Res. , 192 : 122-127 , 1991 ) . ST ( 2 nM ) induces a new P55008 arrest point at 6 h into P55008 . Partial phosphorylation of the retinoblastoma protein was observed at the 2 nM ST arrest point , whereas the retinoblastoma protein was unphosphorylated or underphosphorylated at the 20 nM arrest point . This correlated with the activity of the cyclin-dependent kinase 2 ( P24941 ) and the phosphorylation of the Thr160 residue of p33CDK2 . The cyclin E and cyclin D1/2 levels were reduced at the 20 nM ST arrest point . In HeLa cells that do not arrest in P55008 in response to 2 or 20 nM ST , the retinoblastoma protein and P24941 phosphorylations and P24941 activity were not affected by ST . These results suggest that ST inhibits one or more P55008 -regulating protein kinases , which lie upstream of P24941 . The effect of s-nitroso-glutathione on platelet and leukocyte function during experimental extracorporeal circulation . Treatment with extracorporeal membrane oxygenation ECMO ) is associated with side effects , e.g. , blood cell consumption and activation . Our group has earlier shown that nitric oxide administered as a gas reduces platelet consumption and activation . In the present work we have studied the effect of the NO-donor S-nitroso-glutathione GSNO ) on platelets and leukocytes in an in vitro extracorporeal circuit . Two complete ECMO circuits were perfused with fresh heparinized human blood for 24 hours . GSNO was administered as a continuous infusion to one circuit at a rate of 0.7 mg/hour in four paired experiments and at a rate of 3.5 mg/hour in another four paired experiments . The other circuit was used as a control . Blood samples were withdrawn from both circuits before the start of the experiments and at 0.5 , 1 , 3 , 12 , and 24 hours of perfusion . The samples were analyzed for red blood cell count , leukocyte count , platelet count , platelet membrane expression of glycoproteins GP ) Ib and P08514 /IIIa , leukocyte membrane expression of cluster of differentiation CD ) 11b/ P05107 , as well as plasma concentration of tumor necrosis factor P01375 ) -alpha , interleukin IL ) -1beta , and P10145 . No difference in these parameters between the GSNO and the control circuit at any time point was assayed . In this study , no significant effect of GSNO on circulating platelets or leukocytes during experimental extracorporeal circulation could be shown . Endogenous basic fibroblast growth factor is essential for cyclin E- P24941 activity in multiple external cytokine-induced proliferation of AIDS-associated Kaposi 's sarcoma cells : dual control of AIDS-associated Kaposi 's sarcoma cell growth and cyclin E- P24941 activity by endogenous and external signals . AIDS-associated Kaposi 's sarcoma ( KS ) cell , a key element for development of KS lesions , proliferates in response to external cytokines , such as oncostatin M , the soluble IL-6R- P05231 complex , P01375 , and IL-1beta . In addition , the KS cell-produced basic fibroblast growth factor ( P09038 ) was reported to function as an autocrine growth factor . However , little is known of the exact roles of these external growth factors and endogenous P09038 on proliferation of KS cells , and underlying intracellular events have remained to be defined . We obtained evidence that anti- P09038 Ab abolished growth of KS cells by preventing S phase entry of the cell cycle , even in the presence of the external growth factors . Blockade of the FGF action profoundly inhibited cyclin E expression and cyclin-dependent kinase-2 ( P24941 ) activity , but not D-type cyclin expression and P11802 activity . Exogenously added acidic FGF ( P05230 ) , which generated a rapid tyrosine phosphorylation of P11362 and P21802 on KS cells , reversed the inhibitory effects of anti- P09038 Ab . Thus , FGF actions are essential for cyclin E- P24941 activity and S phase entry . We also observed that the presence of external growth factors markedly induced cyclin E- P24941 activity and S phase entrance , while the addition of P05230 or P09038 alone was insufficient to induce these responses . All this evidence shows that integration of the activities of external growth factors and endogenous P09038 is required for full activation of cyclin E- P24941 activity and KS cell proliferation . Mechanism of oral absorbent DB05269 in lipid abnormalities in experimental uremic rats . BACKGROUND : We have reported that oral sorbent DB05269 ( Q9NRA2 ) is effective in delaying the induction of dialysis in patients with chronic renal failure ( CRF ) because of its effect on lipid metabolism . To clarify the precise mechanism of Q9NRA2 in lipid abnormalities in CRF , we examined the effect of Q9NRA2 on plasma lipid profile , total bile acids ( TBA ) , and lipoprotein lipase ( P06858 ) activity in experimental uremic rats . METHODS : Uremic rats were prepared using male Wistar rats by ligating 5/6 of the renal artery . Uremic rats were randomly divided into two groups as follows : a control group in which rats were maintained on the standard diet and an Q9NRA2 group in which rats were maintained on a diet containing 5 g of Q9NRA2 per 100 g of standard diet for 10 weeks . Plasma P06858 activity was measured as free fatty acid ( FFA ) generation after intravenous administration of heparin . RESULTS : Plasma creatinine at 1.5 +/- 0.1 mg/dl was lower in the Q9NRA2 group than the 1.9 +/- 0.5 mg/ml level in the control group . Q9NRA2 significantly decreased plasma total cholesterol from 192 +/- 29 to 142 +/- 25 mg/dl , triglycerides from 198 +/- 71 to 99 +/- 38 mg/dl , and TBA from 19.6 +/- 2.6 mumol/liter to 8.8 +/- 3.5 mumol/ml . Plasma P06858 activity at 0.22 +/- 0.01 mumol FFA/min/hr was significantly higher in the Q9NRA2 group than 0.15 +/- 0.03 mumol FFA/min/hr in the control group . CONCLUSIONS : These results suggest that Q9NRA2 may improve plasma lipid abnormalities by binding to bile acids in the intestinal lumen and preventing their reabsorption and inhibiting the reduction of P06858 activity in experimental uremic rats . The centrality of P08397 expression levels on DB00855 efficacy . Successful 5-aminolevulinic acid-based photodynamic therapy ( DB00855 ) is dependent on efficient porphyrin synthesis in the inflicted cancer tissue , which is regulated by several enzymes . Irradiation of the tumor excites the light-sensitive porphyrins and results in ROS production and cell death . In this study we investigated the effect of the expression levels of two main enzymes in heme biosynthesis , ALA dehydratase ( P13716 ) and porphobilinogen deaminase ( P08397 ) , on the capacity of K562 cells to undergo cell death following DB00855 . We manipulated P08397 and P13716 expression levels by shRNAs and P08397 overexpressing plasmid . P08397 down-regulation induced an elevation in P13716 activity , while overexpression of P08397 reduced P13716 activity , indicating a novel regulation feedback of P08397 on P13716 activity . This feedback mechanism enabled partial PpIX synthesis under P08397 silencing , whereas P13716 silencing reduced PpIX production to a minimum . DB00855 efficacy was directly correlated to PpIX levels . Thus , only P13716 -silenced cells were not affected by ALA+ irradiation , while following P08397 silencing , the accumulated PpIX , though decreased , was sufficient for successful DB00855 . The alterations in P13716 activity level initiated by changes in P08397 expression indicates P08397 's central role in heme synthesis . This enables efficient DB00855 , even when P08397 is not fully active . Conversely , P13716 loss resulted in reduced PpIX synthesis and consequently failure in DB00855 , due to the absence of compensation mechanism for P13716 . Lupus : novel therapies in clinical development . There have been significant advancements in understanding the immunopathogenesis of systemic lupus erythematosus . However , the developments in therapeutics have been rather slow . DB08879 , a Q9Y275 ( Q9Y275 ) inhibitor has been approved for the treatment of this disease after more than 50 years . Numerous biological agents are being developed which target the B cells , T cells , and various cytokines . Among anti-B cell therapy , drugs target P11836 + cells ( ocrelizumab , SBI-087 ) , P20273 + cells ( epratuzumab ) \or the receptors of tumor necrosis factor ( P01375 ) superfamily ( atacicept , LY2127399 , A-623 ) . Monoclonal antibodies targeting interferon alpha ( IFN-α ) and gamma ( IFN-γ ) and interleukins ( P05231 , 10 ) are being investigated for SLE . Novel targets include toll like receptors , phosphodiesterases , P29965 and retinoid receptors . This review discusses various drugs which are in different phases of clinical trials and hold promise for patients suffering from this chronic debilitating disease . Archaebacterial elongation factor is ADP-ribosylated by diphtheria toxin . Archaebacteria have been defined as a ' third primary kingdom ' of cells in addition to the urkaryotes and the eubacteria . While the latter two correspond approximately to the conventional categories eukaryotes and prokaryotes respectively , the Archaebacteria have up to now comprised four groups of microorganisms : the methanogenic bacteria , the extremely halophilic bacteria and the two thermoacidophilic genera Sulfolobus and Thermoplasma . Based on ribosomal RNA sequence homologies and lipid composition , they apparently form a distinct group . Furthermore they possess or lack typical biochemical markers of both the eukaryotes and the prokaryotes , as well as having unique properties not found elsewhere . Altogether , this indicates that they are not closer to either one of the classical categories . One clear-cut difference between prokaryotes and eukaryotes is the diphtheria toxin reaction , which catalyses the covalent binding of adenosine diphosphate-ribose ( DB02059 ) to the eukaryotic peptide elongation factor P13639 in contrast to the homologous prokaryotic factor EF-G . We report here that diphtheria toxin also catalyses the ADP-riboslation of archaebacterial elongation factors . In this respect , these factors have to be assigned to the P13639 type ; we suppose that the ADP-ribosylatable structure arising so early in evolution is of fundamental importance for the elongation process . Induction of cytokine gene expression in human thyroid epithelial cells irradiated with HZE particles ( iron ions ) . Gene expression profiles were examined using cDNA microarray technology in human thyroid epithelial ( Htori-3 ) cells exposed to a low , non-toxic dose ( 10 cGy ) of radiation from HZE particles in the form of iron ions in the absence or presence of selenomethionine ( SeM ) . A total of 215 genes were differentially regulated 2 h after exposure to a 10-cGy dose of iron-ion radiation . In the microarray analysis , SeM had profound effects on the radiation-induced expression of several specific genes , which includes P00749 , P17936 , P15328 , P15291 and P02452 . Of particular interest to us was a gene cluster , " secreted proteins " , that was up-regulated after radiation exposure . Seven up-regulated genes of this gene cluster fall within the chemokine/cytokine gene cluster , namely , P09341 , P19875 , P05231 , P20809 , P10145 , Q13007 and TGFbeta2 . In microarray studies , the radiation-induced up-regulated expression of some these genes encoding cytokine/chemokine proteins was significantly decreased by SeM treatment . For P10145 , TGFbeta2 , P09341 and P19875 , these observations were validated by qPCR techniques . It is concluded that SeM can regulate ionizing radiation-induced gene expression and may serve as an effective countermeasure for some of the acute inflammatory/immune responses induced by low-dose HZE-particle radiation . Integrative gene network analysis provides novel regulatory relationships , genetic contributions and susceptible targets in autism spectrum disorders . Autism spectrum disorders ( ASDs ) are a group of diseases exhibiting impairment in social drive , communication/language skills and stereotyped behaviors . Though an increased number of candidate genes and molecular interactions have been identified by various approaches , the pathogenesis remains elusive . Based on clinical observations , data from accessible GWAS and expression datasets we identified ASDs gene candidates . Integrative gene network and a novel CNV-centric Node Network ( CNN ) analysis method highlighted ASDs-associated key elements and biological processes . Functional analysis identified neurological functions including synaptic cholinergic receptor ( CHRNA ) families , dopamine receptor ( P14416 ) , and correlations between social behavior and oxytocin related pathways . CNN analysis of genome-wide genetic and expression data identified inheritance-related clusters related to P60484 / Q92574 / Q06787 and mTor/PI3K regulation . Integrative analysis identified potential regulators of networks , specifically P01375 and beta-estradiol , suggesting a potential central role in ASDs . Our data provide information on potential disease mechanisms , and key regulators that may generate novel postulations , and diagnostic molecular biomarkers . Q9Y275 and APRIL expression in B-cell chronic lymphocytic leukemia : correlation with biological and clinical features . B-cell activating factor ( Q9Y275 ) and a proliferation-inducing ligand ( APRIL ) are involved in normal B cell survival and differentiation . We analyzed Q9Y275 and APRIL plasma levels in patients with B-cell chronic lymphocytic leukemia ( B-CLL ) . We also tested intracellular Q9Y275 and APRIL expression in B-CLL and evaluated their prognostic relevance . The percentage of leukemic B cells with intracellular APRIL or Q9Y275 expression correlated significantly with adverse prognostic factors such as Q7Z2W4 -70 and P28907 . Moreover , we found a close relationship between P06858 protein expression or P06858 / Q9UKF5 MFI ratio and proportion of B-CLL cells with intracellular Q9Y275 or APRIL expression . Furthermore , patients with a low percentage of leukemic cells with intracellular Q9Y275 or APRIL expression had a significantly longer overall survival than those with a high proportion of APRIL or Q9Y275 positive leukemic cells . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Concentration-dependent effect of abciximab on platelets and neutrophils in a model of cardiopulmonary bypass . DB00054 is a P08514 /IIIa antagonist used in percutaneous coronary interventions to avoid platelet activation , thrombosis and inflammation . We investigated whether abciximab influenced platelet activation and platelet interaction with neutrophils and polyvinyl chloride ( PVC ) in a cardiopulmonary bypass ( P15086 ) model . Isolated platelets , preincubated with and without 0.1-20 microg/mL of abciximab , were resuspended with neutrophils in plasma and recirculated by a roller pump . Platelet , but not neutrophil adhesion to PVC was inhibited by abciximab . Only high doses of abciximab reduced platelet aggregation size , but simultaneously increased platelet-neutrophil aggregation . DB00054 had no effect on platelet CD62P expression or degranulation , but platelet activation on platelet-neutrophil aggregates increased with high doses . Only low doses inhibited neutrophil degranulation . The concentration-dependent effect of abciximab on platelet-neutrophil interaction reduces its usefulness and stresses the dependency on experimental design in the evaluation of abciximab . Our study does not support the use of abciximab alone in P15086 . However , incorporation of surface-coating the biomaterial with abciximab may be an interesting option . Efficacy of agomelatine , a MT1/ P02795 receptor agonist with P28335 antagonistic properties , in major depressive disorder . Current antidepressants used in major depressive disorder ( MDD ) are still not efficacious enough for many patients due to high levels of treatment resistance and bothersome side-effects . Using a novel blinding method ( interactive voice response system ) , this flexible-dosing study examined the effects of therapeutic doses of agomelatine , a new approach to depressive therapy offering potent melatonergic MT1/ P02795 receptor agonism with P28335 receptor antagonist properties , in patients with moderate-to-severe MDD . This 6-wk , double-blind , parallel-group study randomized 238 patients to 25 mg/d agomelatine ( with dose adjustment at 2 wk to 50 mg/d in patients with insufficient improvement ) or placebo . Depression severity was assessed using the Hamilton Depression Rating Scale ( HAMD ) and the Clinical Global Impression ( CGI ) scale . DB06594 was significantly more efficacious than placebo , with an agomelatine-placebo difference of 3.44 ( p < 0.001 ) using the HAMD final total score . Compared with placebo , agomelatine also had a significant positive impact on CGI - Improvement ( treatment difference=0.45 ) and CGI - Severity ( treatment difference=0.50 ) ( both p=0.006 ) , response rate ( 54.3 % vs. 35.5 % with placebo , p < 0.05 ) and time to first response ( p=0.008 ) . Similar results were seen in patients with the most severe MDD . Depressed mood and sleep items of the HAMD were also significantly improved with agomelatine , which was well tolerated with a safety profile similar to placebo at both doses . This study confirms that agomelatine is effective in treating major depression , including the most severely depressed patients , with a good safety and tolerability profile , therefore providing physicians with an effective pharmacological approach to antidepressant therapy . 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 . DB00158 transport in mouse cumulus-oocyte complexes and preimplantation embryos . Endogenous folate stores are required in preimplantation embryos of several species , but how folates are accumulated and whether they can be replenished has not been determined . Folates are generally taken up into cells by specific transporters , mainly the reduced folate carrier RFC1 ( P41440 protein ) and the high-affinity folate receptors P15328 and P14207 . Quantitative RT-PCR showed that Slc19a1 mRNA was expressed in mouse cumulus-oocyte complexes ( COCs ) and oocytes , whereas Folr1 showed expression only in preimplantation embryos , increasing from the 2-cell stage onward . The mRNAs encoding Folr2 and the intestinal folate transporter Slc46a1 were not detected . DB00563 ( MTX ) , an antifolate often used as a model substrate for folate transport , exhibited saturable transport in COCs and in preimplantation embryos starting at the 2-cell stage . However , folate transport characteristics differed between COCs and embryos . In COCs , transport of MTX and the reduced folate leucovorin was inhibited by the anion transport inhibitor SITS that blocks RFC1 but was insensitive to dynasore , a specific dynamin inhibitor that instead inhibits folate receptor-receptor mediated endocytosis , whereas the opposite was found in 2-cell embryos and blastocysts . The inhibitor profile and transport properties of MTX and leucovorin in COCs correspond to established transport characteristics of RFC1 ( P41440 ) , whereas those in 2-cell embryos and blastocysts correspond with those of P15328 , consistent with the mRNA expression patterns . Considerable folate was accumulated in COCs via RFC1 , but the presence of cumulus cells did not enhance folate accumulation in the enclosed oocyte , indicating a lack of transfer from cumulus to oocyte . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . 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 . Inhibition of tumor necrosis factor signal transduction in endothelial cells by dimethylaminopurine . P01375 ( P01375 ) promotes diverse responses in endothelial cells that are important to the host response to infections and malignancies ; however , less is known of the postreceptor events important to P01375 action in endothelial cells than in many other cell types . Since phosphorylation cascades are implicated in cytokine signaling , the effects of the protein kinase inhibitor dimethylaminopurine ( DMAP ) on P01375 action in bovine aortic endothelial cells ( BAEC ) were investigated . In BAEC , P01375 promotes phosphorylation of eukaryotic initiation factor 4E ( P06730 ) , c-Jun N-terminal kinase ( JNK ) and ceramide-activated protein kinase activities , Jun-b expression , prostacyclin production , and , when protein synthesis is inhibited , cytotoxicity . DMAP abrogated or significantly attenuated each of these responses to P01375 , without affecting the specific binding of P01375 to its receptors . DB11320 , another agent active in the endothelium , promotes phosphorylation of elongation factor-2 ( P13639 ) and prostacyclin production , but not phosphorylation of P06730 in BAEC . DB11320 -stimulated P13639 phosphorylation was not inhibited and prostacyclin production was unaffected by DMAP . These observations demonstrate that a distinct signal transduction cascade , which can be selectively inhibited by DMAP , promotes the response of BAEC to P01375 . Thus , we have identified a reagent , DMAP , that may be useful for characterizing the P01375 signal transduction pathway .

MH_train_1491

MH_train_1491

MH_train_1491

interacts_with DB00015?

multiple_choice

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 . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . 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 . 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 . 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 . [ P11511 inhibitors -- theoretical concept and present experiences in the treatment of endometriosis ] . The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies of endometriosis-associated pain or recurrent disease is primarily aimed at downregulating the ovarian function or at antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is delivering powerful tools for the possible development of new , specific treatment modalities . Recently , aromatase overexpression has been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for conversion of C19 androgens to estrogen in several human tissues . P11511 activity gives rise to local estrogen biosynthesis , which , in turn , stimulates prostaglandin E(2) production by upregulation of cyclooxygenase-2 ( P35354 ) , thus establishing a positive feedback cycle . Another abnormality in endometriosis , i. e. the deficiency in 17 beta-hydroxysteroiddehydrogenase type-II ( 17 beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations collectively favour accumulation of increasing amounts of local estradiol and prostaglandin E(2) in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance , and even invasiveness . Consequently , aromatase and P35354 are promising new therapeutic targets . In summary , specific aromatase inhibitors ( such as Letrozole , DB01217 or Exemestan ) or selective P35354 inhibitors ( e.g. Celecoxib , DB00533 ) are of great interest to be studied in clinical trials in premenopausal woman with endometriosis to extend the spectrum of currently available treatment options . 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 . Inhibition of hydroxymethylglutaryl-coenzyme a reductase reduces Th1 development and promotes Th2 development . Several prospective clinical studies have indicated that hydroxymethylglutaryl-coenzyme A reductase inhibitors , statins , prevent cardiovascular events in part through their antiinflammatory properties . Because inflammation is positively and negatively regulated by T helper ( Th ) 1 cells and Th2 cells , respectively , we examined the effects of statins on the Th polarization in vitro and in vivo . Here we demonstrated that the statins tested , ie , cerivastatin , simvastatin , lovastatin , and atorvastatin , promoted Th2 polarization through both inhibition of Th1 development and augmentation of Th2 development of P01730 + T cells primed in vitro with anti-CD3 antibody and splenic antigen-presenting cells . DB00439 exerted most potent effect on modulation of Th1/Th2 development , and the effect was completely abrogated by an addition of mevalonate . Consistent with in vitro experiments , cerivastatin treatment decreased P01579 production of lymph node cells from mice immunized with ovalbumin emulsified in complete Freund 's adjuvant , indicating that Th1 development is also suppressed in an in vivo proinflammatory environment . In this murine model , cerivastatin significantly reduced mesangial matrix expansion of glomeruli in the kidney and attenuated proteinuria . The decrease of glomerular sclerosis by cerivastatin treatment was positively related to the suppression of interferon ( IFN ) -gamma-producing Th1 response in draining lymph node cells . Hence , these findings strongly suggest that statins ' inhibition of P04035 regulates Th1/Th2 polarization in vivo and such a mechanism possibly plays a pathophysiological role in immune-related glomerular injury . P00747 activator inhibitor-1 promotes inflammatory process induced by cigarette smoke extraction or lipopolysaccharides in alveolar epithelial cells . P00747 activator inhibitor-1 ( P05121 ) plays a role in regulating levels of some cytokines and cell migration in addition to its classic role in inhibiting fibrinolysis . P05121 levels in induced-sputum of chronic obstructive pulmonary disease ( P48444 ) patients were elevated significantly and correlated negatively with pulmonary function . To study the role of P05121 in inflammatory process in P48444 , the authors transfected alveolar epithelial cells ( AECs ) with siRNA-targeted P05121 and stimulated the cells by cigarette smoke extraction ( CSE ) or lipopolysaccharides ( LPS ) . The expression of inflammatory factors , interleukin-8 ( P10145 ) and leukotriene B4 ( LTB4 ) , and the monocyte migration were detected . The exposure to CSE or LPS induced the expression of P05121 , P10145 , and LTB4 in AECs and monocyte migration to AECs . However , they were attenuated after transfection with siRNA-targeted P05121 . In conclusion , P05121 stimulates inflammation induced by CSE or LPS in AECs through up-regulation of inflammatory factors and promoting inflammatory cell migration . P05121 may play a proinflammatory role in the pathogenesis of P48444 . [ 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 . Transgenic GATA-4 expression induces adrenocortical tumorigenesis in C57Bl/6 mice . A link between elevated luteinizing hormone ( LH ) levels , GATA-4 and LH receptor ( P22888 ) expression and gonadotropin-dependent adrenocortical tumorigenesis in humans and mice has been shown . To assess the mechanistic tumorigenic interrelationships between these factors , we transgenically expressed Gata4 under the 21-hydroxylase promoter ( Cyp21a1 , 21-OH ) in C57Bl/6N mice . There was a gradual age-dependent increase of GATA-4 expression only in 21-OH-GATA-4 ( TG ) female adrenals , in association with slowly progressing neoplasia of non-steroidogenic spindle-shaped A cells in the subcapsular cortex . Gonadectomy ( P11441 ) , apparently through direct action of elevated serum LH , markedly enhanced the adrenocortical neoplasia , which now also appeared in P11441 TG males . The neoplastic areas of the post- P11441 TG adrenals contained , besides A cells , larger lipid-laden , steroidogenically active and P22888 -positive B cells . Prolonged ( > 10 months ) exposure to elevated post- P11441 LH levels resulted in formation of adrenocortical adenomas in the TG mice . Intact and P11441 TG mouse adrenals displayed elevated Q8WW38 and decreased GATA-6 expression . Additionally , increased expression/activation of components of the Inhbb-Acvr2a-Acvr1c- Q15796 /3 signaling system was observed in 12-month-old P11441 TG adrenals . Our findings show that two distinct GATA-4-dependent populations of neoplastic adrenocortical cells form : non-steroidogenic LH-independent A cells and steroidogenic LH-dependent B cells . P01308 -like growth factor-1 receptor and phosphorylated AKT-serine 473 expression in 132 resected thymomas and thymic carcinomas . BACKGROUND : Thymic malignancies are rare tumors . The insulin-like growth factor-1 ( DB01277 ) / DB01277 receptor ( IGF-1R ) system is involved in the development of the thymus . IGF-1R expression in thymic epithelial malignancies is unknown . METHODS : The authors investigated the expression of IGF-1R and phosphorylated AKT serine 473 ( p-AKT ) by using immunohistochemistry and examined the clinicopathologic correlations in a retrospective , single-institution surgical series of 132 patients with thymic epithelial malignancies . RESULTS : Earlier disease stage , less aggressive histologic types , and complete resection were significant positive prognostic factors for disease-related survival and progression-free survival , and being a woman was a better prognostic factor for disease-related survival . IGF-1R and p-AKT protein levels were expressed in 20 % and 36 % of thymic tumors , respectively . Both markers were expressed more commonly in recurrent disease than in primary tumors , in more aggressive subtypes , and in more advanced disease stages . There was a trend toward better survival and progression-free survival in patients who were negative for IGF-1R or p-AKT expression in the whole series . When only the 91 primary tumors , P08069 expression was associated with worse progression-free survival ( P < .001 ) . CONCLUSIONS : The current retrospective analysis demonstrated that disease stage , tumor histology , sex , and resection type were major prognostic factors in the survival of patients with thymic malignancies . The expression levels of IGF-1R and p-AKT in thymic tumors suggested that IGF-1R is a potential target for treatment . Role of TGFbeta/Smad signaling in gremlin induction of human trabecular meshwork extracellular matrix proteins . PURPOSE : The bone morphogenic protein ( BMP ) antagonist gremlin is elevated in glaucomatous trabecular meshwork ( TM ) cells and tissues and elevates intraocular pressure ( IOP ) . Gremlin also blocks P12644 inhibition of transforming growth factor ( TGF ) -β2 induction of TM extracellular matrix ( Q13201 ) proteins . The purpose of this study was to determine whether Gremlin regulates Q13201 proteins in cultured human TM cells . METHODS : Human TM cells were treated with recombinant gremlin to determine the effects on Q13201 gene and protein expression . Expression of the Q13201 genes FN , COL1 , P05121 , and P15502 was examined in cultured human TM cells by quantitative RT-PCR and Western immunoblot analysis . TM cells were pretreated with TGFBR inhibitors ( LY364947 , SB431542 or P36897 / P61812 siRNAs ) , inhibitors of the Smad signaling pathway ( SIS3 or Q15796 /3/4 siRNAs ) , or P29279 siRNA to identify the signaling pathway(s) involved in gremlin induction of Q13201 gene and protein expression . RESULTS : All Q13201 genes analyzed ( FN , COL1 , P05121 , and P15502 ) were induced by gremlin . This gremlin induction of Q13201 genes and protein expression was blocked by inhibitors of TGFBR and the canonical Q15796 /3/4 and P29279 signaling pathways . CONCLUSIONS : Gremlin employs canonical TGFβ2/Smad signaling to induce Q13201 genes and proteins in cultured human TM cells . Gremlin also induces both TGFβ2 and P29279 , which can act downstream to mediate some of these Q13201 changes in TM cells . 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 . DB04956 . Tumour necrosis factor-alpha ( P01375 ) is well established as a key mediator in the inflammatory response seen in various disease processes including sepsis . P01375 is involved in virtually all features of septic shock and multiple organ failure . Anti- P01375 strategies are thus appealing and have been effective at reducing inflammation and morbidity in certain conditions including rheumatoid arthritis and Crohn 's disease . DB04956 is the F(ab')2 fragment of a murine anti- P01375 antibody , and has been evaluated in clinical trials in septic patients . The results suggest that the drug is well tolerated , and may be of benefit in certain groups of patients with sepsis . A large , randomised , clinical trial of afelimomab in patients with severe sepsis has recently been completed and the results are eagerly awaited . More work is necessary to identify a means of selecting which patients are most likely to benefit from this type of therapy in sepsis . 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 . 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 . P22888 splicing variants in bovine Leydig cells . The luteinizing hormone receptor ( LHR ) plays a key role in testosterone production through its interaction with the gonadotropins , LH and chorionic gonadotropin . We examined the LHR splicing pattern in bovine Leydig cells ; LH-induced expression of eight cloned splicing variants was detected by real-time PCR . DB00044 applied to cultured Leydig cells resulted in expression of full-length LHR and the A and B isoforms , as well as secretion of testosterone , which first increased , then declined , and then increased further , with increased LH levels . The secretion of testosterone progressively increased with increasing LH , but the expression levels of LHR ( FL , A , and B ) did not increase correspondingly . We conclude that the LHR splicing pattern is complex in bovine Leydig cells , and that expression of full-length LHR and isoforms A and B changes when induced with LH .

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interacts_with DB00333?

multiple_choice

3D Q99707 measurement : from 1.5 T to 3.0 T . This study investigates some of the issues involved in magnetization transfer ratio ( Q99707 ) acquisition , and in particular aims to determine whether high quality in vivo Q99707 measurements can be made at 3.0 T . The dependency of the Q99707 white-to-grey matter contrast to noise ratio ( P21554 ) on MT pulse characteristics at 1.5 T and at 3.0 T was investigated using an established two-pool model for MT . The simulations showed that MT pulse parameters optimizing the P21554 can be derived for both field strengths . Both the SNR and the P21554 of Q99707 maps at 3.0 T were increased compared to 1.5 T . Images obtained using a safe in vivo Q99707 acquisition protocol based on results of simulations at 3.0 T are presented . SAR131675 , a potent and selective P35916 -TK inhibitor with antilymphangiogenic , antitumoral , and antimetastatic activities . SAR131675 is a potent and selective P35916 inhibitor . It inhibited P35916 tyrosine kinase activity and P35916 autophosphorylation in P29320 cells with IC(50) values of 20 and 45 nmol/L , respectively . SAR131675 dose dependently inhibited the proliferation of primary human lymphatic cells , induced by the P35916 ligands P49767 and O43915 , with an IC(50) of about 20 nmol/L . SAR131675 was found to be highly selective for P35916 versus 107 receptors , enzymes , ion channels , and 65 kinases . However , it was moderately active on P35968 with a P35916 / P35968 ratio of about 10 . SAR131675 had no antiproliferative activity on a panel of 30 tumors and primary cells , further showing its high specificity and indicating that SAR131675 is not a cytotoxic or cytostatic agent . SAR131675 was very well tolerated in mice and showed a potent antitumoral effect in several orthotopic and syngenic models , including mammary 4T1 carcinoma and Q13546 .Tag2 tumors . Interestingly , it significantly reduced lymph node invasion and lung metastasis , showing its antilymphangiogenic activity in vivo . Moreover , treatment of mice before resection of 4T1 primary tumors was sufficient to prevent metastasis . Tumor-associated macrophages ( TAM ) play an important role in tumor growth and metastasis . The expression of P35916 on TAMs has been recently described . F4/80 immunostaining clearly showed that SAR131675 significantly reduced TAM infiltration and aggregation in 4T1 tumors . Taken together , SAR131675 is the first highly specific P35916 -TK inhibitor described to date , displaying significant antitumoral and antimetastatic activities in vivo through inhibition of lymphangiogenesis and TAM invasion . Q07869 -γ regulates carnitine homeostasis and mitochondrial function in a lamb model of increased pulmonary blood flow . OBJECTIVE : DB00583 homeostasis is disrupted in lambs with endothelial dysfunction secondary to increased pulmonary blood flow ( Shunt ) . Our recent studies have also indicated that the disruption in carnitine homeostasis correlates with a decrease in Q07869 -γ expression in Shunt lambs . Thus , this study was carried out to determine if there is a causal link between loss of Q07869 -γ signaling and carnitine dysfunction , and whether the Q07869 -γ agonist , rosiglitazone preserves carnitine homeostasis in Shunt lambs . METHODS AND RESULTS : siRNA-mediated Q07869 -γ knockdown significantly reduced carnitine palmitoyltransferases 1 and 2 ( CPT1 and 2 ) and carnitine acetyltransferase ( P43155 ) protein levels . This decrease in carnitine regulatory proteins resulted in a disruption in carnitine homeostasis and induced mitochondrial dysfunction , as determined by a reduction in cellular DB00171 levels . In turn , the decrease in cellular DB00171 attenuated NO signaling through a reduction in P29474 /Hsp90 interactions and enhanced P29474 uncoupling . In vivo , rosiglitazone treatment preserved carnitine homeostasis and attenuated the development of mitochondrial dysfunction in Shunt lambs maintaining DB00171 levels . This in turn preserved P29474 /Hsp90 interactions and NO signaling . CONCLUSION : Our study indicates that Q07869 -γ signaling plays an important role in maintaining mitochondrial function through the regulation of carnitine homeostasis both in vitro and in vivo . Further , it identifies a new mechanism by which Q07869 -γ regulates NO signaling through Hsp90 . Thus , Q07869 -γ agonists may have therapeutic potential in preventing the endothelial dysfunction in children with increased pulmonary blood flow . P06401 plays a major antiinflammatory role in human myometrial cells by antagonism of nuclear factor-kappaB activation of cyclooxygenase 2 expression . Spontaneous labor in women and in other mammals is likely mediated by a concerted series of biochemical events that negatively impact the ability of the progesterone receptor ( PR ) to regulate target genes that maintain myometrial quiescence . In the present study , we tested the hypothesis that progesterone/PR inhibits uterine contractility by blocking nuclear factor kappaB ( NF-kappaB ) activation and induction of cyclooxygenase-2 ( P35354 ) , a contractile gene that is up-regulated in labor . To uncover mechanisms for regulation of uterine P35354 , immortalized human fundal myometrial cells were treated with IL-1beta +/- progesterone . IL-1beta alone caused a marked up-regulation of P35354 mRNA , whereas treatment with progesterone suppressed this induction . This was also observed in human breast cancer ( T47D ) cells . In both cell lines , this inhibitory effect of progesterone was blocked by DB00834 . Using chromatin immunoprecipitation , we observed that IL-1beta stimulated recruitment of NF-kappaB p65 to both proximal and distal NF-kappaB elements of the P35354 promoter ; these effects were diminished by coincubation with progesterone . The ability of progesterone to inhibit P35354 expression in myometrial cells was associated with rapid induction of mRNA and protein levels of inhibitor of kappaBalpha , a protein that blocks NF-kappaB transactivation . Furthermore , small interfering RNA-mediated ablation of both PR-A and PR-B isoforms in T47D cells greatly enhanced NF-kappaB activation and P35354 expression . These effects were observed in the absence of exogenous progesterone , suggesting a ligand-independent action of PR . Based on these findings , we propose that PR may inhibit NF-kappaB activation of P35354 gene expression and uterine contractility via ligand-dependent and ligand-independent mechanisms . Population pharmacokinetic study of memantine : effects of clinical and genetic factors . BACKGROUND AND OBJECTIVE : Memantine , a frequently prescribed anti-dementia drug , is mainly eliminated unchanged by the kidneys , partly via tubular secretion . Considerable inter-individual variability in plasma concentrations has been reported . We aimed to investigate clinical and genetic factors influencing memantine disposition . METHODS : A population pharmacokinetic study was performed including data from 108 patients recruited in a naturalistic setting . Patients were genotyped for common polymorphisms in renal cation transporters ( O15245 /2/5 , Q96FL8 , P08183 ) and nuclear receptors ( O75469 , Q14994 , RXR , Q07869 ) involved in transporter expression . RESULTS : The average clearance was 5.2 L/h with a 27 % inter-individual variability ( percentage coefficient of variation ) . Glomerular filtration rate ( p = 0.007 ) and sex ( p = 0.001 ) markedly influenced memantine clearance . O75469 rs1523130 was identified as the unique significant genetic covariate for memantine clearance ( p = 0.006 ) , with carriers of the O75469 rs1523130 CT/TT genotypes presenting a 16 % slower memantine elimination than carriers of the CC genotype . CONCLUSION : The better understanding of inter-individual variability of memantine disposition might be beneficial in the context of individual dose optimization . Mice deficient in phosphodiesterase-4A display anxiogenic-like behavior . RATIONALE : Phosphodiesterases ( PDEs ) are a super family of enzymes responsible for the halting of intracellular cyclic nucleotide signaling and may represent novel therapeutic targets for treatment of cognitive disorders . DB05876 is of considerable interest to cognitive research because it is highly expressed in the brain , particularly in the cognition-related brain regions . Recently , the functional role of Q07343 and Q08499 , two of the four DB05876 subtypes ( P27815 , B , C , and D ) , in behavior has begun to be identified ; however , the role of P27815 in the regulation of behavior is still unknown . OBJECTIVES : The purpose of this study was to characterize the functional role of P27815 in behavior . METHODS : The role of P27815 in behavior was evaluated through a battery of behavioral tests using P27815 knockout ( KO ) mice ; urine corticosterone levels were also measured . RESULTS : P27815 KO mice exhibited improved memory in the step-through-passive-avoidance test . They also displayed anxiogenic-like behavior in elevated-plus maze , holeboard , light-dark transition , and novelty suppressed feeding tests . Consistent with the anxiety profile , P27815 KO mice had elevated corticosterone levels compared with wild-type controls post-stress . Interestingly , P27815 KO mice displayed no change in object recognition , Morris water maze , forced swim , tail suspension , and duration of anesthesia induced by co-administration of xylazine and ketamine ( suggesting that P27815 KO may not be emetic ) . CONCLUSIONS : These results suggest that P27815 may be important in the regulation of emotional memory and anxiety-like behavior , but not emesis . P27815 could possibly represent a novel therapeutic target in the future for anxiety or disorders affecting memory . 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 . Genetics of physical activity and physical inactivity in humans . Emerging evidence suggests that physical activity and sedentary behavior [ reflected in physical inactivity ( PI ) ] , might be two different phenotypes that may have distinct underlying physiological mechanisms . The purpose of this review is to summarize the existing literature on the genetic determinants of PA and PI phenotypes in humans , considering them as distinct behaviors . Completed in January 2011 , this review includes family studies , twin studies , association studies , genome-wide linkage studies and genome-wide association scan ( GWAs ) reporting different physical activity/inactivity-related phenotypes . In regards to PA , familial aggregation studies resulted in heritability estimates ranging from 0 to 60 % , and twin studies yielded heritability estimates ( a(2) ) and shared environment ( c(2) ) scores for PA phenotypes ranging from 0.00 to 0.85 and 0.00 to 0.84 , respectively . Unique environmental ( e(2) ) results are well dispersed from 0.12 to 0.72 . Suggestive linkages were found with markers nearby different activity-related genes : P24530 , P32245 , P25874 , P12104 , P41180 , Q8IVB4 . Significant associations with PA phenotypes were found for Ace , Gln223ARrg , P32245 and P14416 genes . We found one GWAs that reported novel SNPs in the O95340 gene on chromosome 10q23.2 and in two intergenic regions on chromosomes 2q33.1 and 18p11.32 . Heritability estimates for PI ranged from 25 to 60 % and linkage studies recorded higher LOD scores for PI versus PA . The P12821 genotype was strongly associated with PI . There are potentially different genetic influences on PA versus PI phenotypes . Future studies should focus on the different genetic influences on PA and PI to improve our understanding of underlying determinants of these behaviors . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Characterization of the activity of the PI3K/ P42345 inhibitor DB05241 ( SAR245409 ) in tumor models with diverse genetic alterations affecting the PI3K pathway . Activation of the PI3K ( phosphoinositide 3-kinase ) pathway is a frequent occurrence in human tumors and is thought to promote growth , survival , and resistance to diverse therapies . Here , we report pharmacologic characterization of the pyridopyrimidinone derivative DB05241 ( SAR245409 ) , a potent and highly selective pan inhibitor of class I PI3Ks ( α , β , γ , and δ ) with activity against P42345 . Broad kinase selectivity profiling of > 130 protein kinases revealed that DB05241 is highly selective for class I PI3Ks and P42345 over other kinases . In cellular assays , DB05241 inhibits the formation of PIP(3) in the membrane , and inhibits phosphorylation of AKT , p70S6K , and S6 phosphorylation in multiple tumor cell lines with different genetic alterations affecting the PI3K pathway . In a panel of tumor cell lines , DB05241 inhibits proliferation with a wide range of potencies , with evidence of an impact of genotype on sensitivity . In mouse xenograft models , oral administration of DB05241 results in dose-dependent inhibition of phosphorylation of AKT , p70S6K , and S6 with a duration of action of approximately 24 hours . Repeat dose administration of DB05241 results in significant tumor growth inhibition in multiple human xenograft models in nude mice that is associated with antiproliferative , antiangiogenic , and proapoptotic effects . Overexpression of the μ-opioid receptor in human non-small cell lung cancer promotes Akt and P42345 activation , tumor growth , and metastasis . BACKGROUND : Recent epidemiologic studies suggesting that there were differences in cancer recurrence contingent on anesthetic regimens have raised the possibility that μ-opioid agonists can influence cancer progression . Based on our previous studies indicating the μ-opioid receptor ( MOR ) is up-regulated in several types of non-small cell lung cancer , this study examined the functional significance of MOR overexpression to elucidate a possible mechanism for the epidemiologic findings . METHODS : Stable vector control and P35372 overexpressing human bronchioloalveolar carcinoma cells were evaluated using immunoblot analysis , proliferation and transendothelial extravasation assays with or without Akt inhibitor , P42345 inhibitor ( temsirolimus ) , or the peripheral MOR antagonist , methylnaltrexone . In human lung cancer xenograft models , primary tumor growth rates and lung metastasis were analyzed using consecutive tumor volume measurements and nestin immunoreactivity in lungs of the nude mouse model . RESULTS : The authors provide evidence that MOR is an important regulator of lung cancer progression . MOR overexpression increased Akt and P42345 activation , proliferation , and extravasation in human bronchioloalveolar carcinoma cells . In vivo , overexpression of MOR in human bronchoalveolar carcinoma cells increased primary tumor growth rates in nude mice by approximately 2.5-fold and lung metastasis by approximately 20-fold compared with vector control cells ( n = 4 per condition ) . CONCLUSIONS : The overexpression data suggest a possible direct effect of MOR on Akt and P42345 activation and lung cancer progression . Such an effect provides a plausible explanation for the epidemiologic findings . The authors ' observations further suggest that exploration of MOR in non-small cell lung carcinoma merits further study both as a diagnostic and therapeutic option . DB00115 responsive homocystinuria and megaloblastic anemia : heterogeneity in methylcobalamin deficiency . A male infant with methyl-B12 deficiency ( cblE ) presented at age 6 weeks with lethargy , staring spells , and vomiting . He later became hypotonic and unresponsive to stimuli and required intubation and ventilation . He had homocystinuria and hypomethioninemia with megaloblastic anemia but normal serum folate and vitamin B12 concentrations . No methylmalonic aciduria was detected . Fibroblasts , cultured from the patient , were unable to grow in medium in which homocysteine replaced methionine and incorporated abnormally small amounts of [ 14C ] -methyl- DB00116 but normal amounts of [ 14C ] -propionate into protein . Methyl-B12 content of fibroblasts was low , while the adenosyl-B12 content was normal . Q99707 activity was decreased when the assay was performed under both optimal and suboptimal reducing conditions , suggesting heterogeneity in the cblE disease . The patient responded dramatically to hydroxocobalamin treatment . Homocystinuria disappeared after 10 days of therapy , and methionine was normalized after 3 weeks . Psychometric testing at age 15 months showed a developmental age of 9 months . ZNF804a regulates expression of the schizophrenia-associated genes Q9NQE7 , P21964 , Q07343 , and P14416 . ZNF804a was identified by a genome-wide association study ( GWAS ) in which a single nucleotide polymorphism ( SNP rs1344706 ) in ZNF804a reached genome-wide statistical significance for association with a combined diagnosis of schizophrenia ( SZ ) and bipolar disorder . Although the molecular function of ZNF804a is unknown , the amino acid sequence is predicted to contain a C2H2-type zinc-finger domain and suggests ZNF804a plays a role in DNA binding and transcription . Here , we confirm that ZNF804a directly contributes to transcriptional control by regulating the expression of several SZ associated genes and directly interacts with chromatin proximal to the promoter regions of Q9NQE7 and P21964 , the two genes we find upregulated by ZNF804a . Using immunochemistry we establish that ZNF804a is localized to the nucleus of rat neural progenitor cells in culture and in vivo . We demonstrate that expression of ZNF804a results in a significant increase in transcript levels of Q9NQE7 and P21964 , relative to GFP transfected controls , and a statistically significant decrease in transcript levels of Q07343 and P14416 . Furthermore , we show using chromatin immunoprecipitation assays ( ChIP ) that both epitope-tagged and endogenous ZNF804a directly interacts with the promoter regions of Q9NQE7 and P21964 , suggesting a direct upregulation of transcription by ZNF804a on the expression of these genes . These results are the first to confirm that ZNF804a regulates transcription levels of four SZ associated genes , and binds to chromatin proximal to promoters of two SZ genes . These results suggest a model where ZNF804a may modulate a transcriptional network of SZ associated genes . 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 . Novel structural features of CDK inhibition revealed by an ab initio computational method combined with dynamic simulations . The rational development of specific inhibitors for the approximately 500 protein kinases encoded in the human genome is impeded by a poor understanding of the structural basis for the activity and selectivity of small molecules that compete for DB00171 binding . Combining classical dynamic simulations with a novel ab initio computational approach linear-scalable to molecular interactions involving thousands of atoms , we have investigated the binding of five distinct inhibitors to the cyclin-dependent kinase P24941 . We report here that polarization and dynamic hydrogen bonding effects , so far undetected by crystallography , affect both their activity and selectivity . The effects arise from the specific solvation patterns of water molecules in the DB00171 binding pocket or the intermittent formation of hydrogen bonds during the dynamics of CDK/inhibitor interactions and explain the unexpectedly high potency of certain inhibitors such as 3-(3H-imidazol-4-ylmethylene)-5-methoxy-1,3-dihydro-indol-2-one ( DB03428 ) . The Lys89 residue in the DB00171 -binding pocket of P24941 is observed to form temporary hydrogen bonds with the three most potent inhibitors . This residue is replaced in P11802 by Thr89 , whose shorter side-chain can not form similar bonds , explaining the relative selectivity of the inhibitors for P24941 . Our results provide a generally applicable computational method for the analysis of biomolecular structures and reveal hitherto unrecognized features of the interaction between protein kinases and their inhibitors . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . Selective inhibitors of Q02750 /ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells . The nonsteroidal anti-inflammatory drug ( NSAID ) sulindac prevents experimental colon cancer and can regress precancerous polyps in humans . Sulindac sulfide inhibits cyclooxygenase ( P36551 ) -mediated prostaglandin synthesis and retards the growth of cultured colon cell lines primarily by inducing apoptosis . Given the known role of mitogen-activated protein kinase ( MAPK ) in signal transduction and the regulation of cell survival and death , we determined the effect of sulindac sulfide on MAPK activation , P35354 expression , and apoptosis induction in HCA-7 human colon cancer cells . Sulindac sulfide treatment was associated with activation of ERKp44/42 and p38 MAPK in a dosage- and time-dependent manner , and also activated upstream MEK . Similar results were seen in HCT-15 cells and also with the selective P35354 inhibitor NS398 . ERKp44/42 and p38 activation were accompanied by an induction of P35354 protein expression . Selective inhibitors of sulindac sulfide-induced ERKp44/42 ( PD98059 ) and p38 MAPK ( SB203580 ) activation also suppressed the induction of P35354 by this NSAID . Furthermore , both MAPK inhibitors significantly augmented sulindac sulfide-induced apoptosis , as did suppression of constitutive P35354 using antisense oligonucleotides . In conclusion , MEK/ P29323 and p38 MAPK activation mediate P35354 induction by sulindac sulfide . Selective inhibitors of these MAPKs potentiate apoptosis induction by this NSAID , suggesting a novel strategy for the prevention or treatment of colorectal cancer . Expression of two variants of the human mu opioid receptor mRNA in SK-N-SH cells and human brain . A partial mu opioid receptor gene was isolated from a human genomic library using a mouse delta opioid receptor cDNA as a probe . Using information from this genomic clone and the published human mu receptor , P35372 , a cDNA was isolated from SK-N-SH mRNA that codes for a variant of the P35372 mRNA , MOR1A . The presence of MOR1A is also shown in human brain using RT-PCR . MOR1A differs from P35372 in that the 3' terminal intron has not been removed . An in-frame termination codon is found four amino acids after the 5' consensus splice site , making MOR1A eight amino acids shorter than P35372 . Both receptors show similar ligand binding and coupling to DB02527 in CHO- P04264 cells . The C-terminal differences between P35372 and MOR1A could have effects on receptor coupling or receptor transport and localization . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Wnt1 and Q02750 cooperate to promote cyclin D1 accumulation and cellular transformation . Members of the Wnt family of signal transducers regulate cellular differentiation/reorganization and cellular proliferation . However , few pro-proliferative targets of Wnt have been identified . We now show that cyclin D1 , a critical mediator of cell cycle progression , is a downstream target of Wnt-dependent signaling . NIH-3T3 cell lines engineered to overexpress Wnt1 displayed reduced glycogen synthase kinase-3beta activity . Wnt1-dependent glycogen synthase kinase-3beta inhibition corresponded with decreased cyclin D1 proteolysis and , thus , hyperaccumulation of active cyclin D1. P11802 ( cyclin-dependent kinase 4 ) kinase . However , in the absence of serum-derived growth factors , Wnt-1 was not sufficient to drive cyclin D1 accumulation or S-phase entry . In contrast , cells engineered to co-express Wnt1 and activated Q02750 accumulated high levels of cyclin D1 and entered the DNA synthetic phase in the absence of serum-derived growth factors , a characteristic of neoplastic transformation . The ability of a dominant-negative cyclin D1 mutant , D1-T156A , to inhibit Wnt1/ Q02750 -dependent S-phase entry suggests that cyclin D1 is a critical downstream target for Wnt1- and Q02750 -dependent cellular proliferation . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . 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 .

MH_train_1493

MH_train_1493

MH_train_1493

interacts_with DB01285?

multiple_choice

Reduced neuronal injury after treatment with NG-nitro-L-arginine methyl ester ( L-NAME ) or 2-sulfo-phenyl-N-tert-butyl nitrone ( S-PBN ) following experimental brain contusion . OBJECTIVE : DB00435 ( NO ) and oxygen free radicals are implicated in the pathophysiology of traumatic brain injury ( TBI ) . Peroxynitrite formation from NO and superoxide contributes to secondary neuronal injury but the neuroprotective effects of nitric oxide synthase ( NOS ) -inhibitors have been contradictory . This study was undertaken to examine whether PTtic administration of the ( NOS ) -inhibitor DB04223 methyl ester ( L-NAME ) , and a combination of L-NAME and the nitrone radical scavenger 2-sulfo-phenyl-N-tert-butyl nitrone ( S-PBN ) favorable affects neuronal injury in a model of TBI . METHODS : A weight-drop model of TBI was used . The animals received L-NAME , S-PBN or a combination of the drugs 15 minutes prothrombin time ( PT ) and sacrificed after 24 hours or six days . NOS activity was measured by the conversion of L-[U-C]arginine to L-[U-C]citrulline . Peroxynitrite formation , cellular apoptosis , neuronal degeneration and survival were assessed by nitrotyrosine- , TUNEL- , Fluoro-Jade- and NeuN-stainings . RESULTS : P29474 and P29475 activity was significantly reduced in animals that received L-NAME alone or the combination with S-PBN. P35228 activity or P35228 immunoreactivity was not affected . All treatments significantly reduced neuronal degeneration and nitrotyrosine immunoreactivity at 24 hours and increased neuronal survival at six days PT . No differences were detected between L-NAME and L-NAME + S-PBN groups . CONCLUSION : NO from NOS contributes to secondary neuronal injury in this TBI-model . PTtic treatment does not inhibit early beneficial NO-related effects . L-NAME and S-PBN limit peroxynitrite formation , promoting neuronal survival . The combination of L-NAME and S-PBN was neuroprotective ; surprisingly no additive effects were found on nitrotyrosine formation , apoptosis or neuronal survival . DB01285 -releasing Factor Changes the Phenotype and Function of Dendritic Cells in Mouse Mesenteric Lymph Nodes . BACKGROUND/AIMS : Dendritic cells ( DCs ) are a significant contributor to the pathology of numerous chronic inflammatory autoimmune disorders ; however , the effects of P06850 ( CRF ) on intestinal DCs are poorly understood . In this study , we investigated the role of CRF in alterations of intestinal dendritic cell phenotype and function . METHODS : Mouse mesenteric lymph node dendritic cells ( MLNDCs ) were obtained using magnetic bead sorting . Surface expression of CRF receptor type 1 ( P34998 ) and Q13324 was determined by double-labeling immunofluorescence and quantitative polymerase chain reaction ( qPCR ) and MLNDCs were subsequently exposed to CRF in the presence or absence of P34998 and Q13324 antagonists . Expression of surface molecules ( MHC-I and MHC-II ) and co-stimulatory molecules ( P33681 and P42081 ) was determined by flow cytometric and western blot analyses , and the T cell stimulatory capacity of MLNDCs was evaluated by mixed lymphocyte reaction . RESULTS : Immunofluorescent staining and quatitative polymerase chain reaction indicated that both the CRF receptors ( P34998 and CRF-2 ) are expressed on the surface of MLNDCs . Exposure to CRF increased the expression of MHC-II on MLNDCs as well as their capacity to stimulate T cell proliferation . MLNDCs treated with P34998 antagonist exhibited a phenotype characterized by a less activated state and reduced surface expression of MHC-II , and consequently showed reduced capacity to stimulate T cells . In contrast , treatment of MLNDCs with Q13324 antagonist yielded an opposite result . CONCLUSIONS : CRF can alter the phenotype and function of intestinal DCs through direct action on P34998 and Q13324 , and activation of the P34998 and Q13324 pathways yields opposing outcomes . 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 . DB05595 in lung cancer . DB00158 is essential for proliferating cells and folate transport pathways and folate-dependent metabolic processes show promise as targets for anti-neoplastic therapy . DB00158 receptor α ( P15328 ) , a folate transporter , is an attractive target for anti-neoplastic therapy due to its high affinity for folate , restricted range of expression in normal tissue and differential over-expression in malignant tissue . P15328 is expressed in non-small cell lung cancer , with a higher expression in adenocarcinoma compared with squamous cell carcinoma . DB05595 is a monoclonal antibody targeting P15328 which in pre-clinical studies led to cytotoxicity of P15328 -expressing cells , inhibited tumor growth in animal models and showed limited reactivity with normal tissue . In phase I/II trials , farletuzumab was well tolerated as a single-agent and in combination , without additive toxicity with chemotherapy . An ongoing phase II , double blind , placebo-controlled study is evaluating farletuzumab in patients with P15328 expressing metastatic adenocarcinoma of lung . DB01285 releasing factor receptor 1 ( CRF1 ) and CRF2 agonists exert an anti-inflammatory effect during the early phase of inflammation suppressing LPS-induced P01375 release from macrophages via induction of P35354 and DB00917 . P06850 ( CRF ) , the principal regulator of the hypothalamus-pituitary-adrenal ( Q9Y251 ) axis , also modulates the inflammatory response directly , via its effect on mast cells and macrophages . On macrophages , it augments production of lipopolysaccharide ( LPS ) -induced pro-inflammatory cytokines . CRF and its related peptides may also act as anti-inflammatory agents . Aim of the present work was to examine the role of macrophages on the anti-inflammatory effects of CRF-peptides and the mechanism involved . Thus , we examined if CRF receptor 1 ( CRF1 ) and CRF2 agonists exert any anti-inflammatory effect on primary mouse macrophages . We have found that : ( a ) CRF , P55089 ( P55089 )1 and Q96RP3 transiently suppressed the release of Tumor Necrosis Factor-alpha ( P01375 ) in LPS-activated macrophages , an effect peaking at 4 h . This effect did not involve changes on P01375 transcription . ( b ) CRF peptide-induced suppression of P01375 release depended on induction of P35354 and DB00917 synthesis . ( c ) Use of specific CRF1 and CRF2 antagonists suggested that this effect involved both CRF receptor types . ( d ) The effect of CRF-peptides on P35354 was mediated via PI3K and p38MAPK . ( e ) Longer exposure of macrophages to CRF-peptides resulted in induction of P01375 production via enhancement of its transcription . In conclusion , this is the first report suggesting that CRF1 and CRF2 agonists exert a biphasic effect on macrophages . During the early stages of the inflammatory response , they suppress P01375 release via induction of P35354 / DB00917 while later on they induce P01375 transcription . Hence , the reported anti-inflammatory effect of CRF-peptides appears to involve macrophages and is confined at the early stage of inflammation . Telmisartan , its potential therapeutic implications in cardiometabolic disorders . There is a growing body of evidence that the renin-angiotensin system ( DB01367 ) plays a pivotal role in the pathogenesis of cardiovascular diseases . Indeed , large clinical trials have demonstrated substantial benefit of the blockade of this system for cardiovascular-organ protection . 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 . Telmisartan will be a promising cardiometabolic sartan due to its unique peroxisome proliferator-activated receptor-gamma ( P37231 ) -inducing properties as well . In this review , we focused on telmisartan , and discussed its potential therapeutic implications in cardiometabolic disorders . Induction of cytokine gene expression in human thyroid epithelial cells irradiated with HZE particles ( iron ions ) . Gene expression profiles were examined using cDNA microarray technology in human thyroid epithelial ( Htori-3 ) cells exposed to a low , non-toxic dose ( 10 cGy ) of radiation from HZE particles in the form of iron ions in the absence or presence of selenomethionine ( SeM ) . A total of 215 genes were differentially regulated 2 h after exposure to a 10-cGy dose of iron-ion radiation . In the microarray analysis , SeM had profound effects on the radiation-induced expression of several specific genes , which includes P00749 , P17936 , P15328 , P15291 and P02452 . Of particular interest to us was a gene cluster , " secreted proteins " , that was up-regulated after radiation exposure . Seven up-regulated genes of this gene cluster fall within the chemokine/cytokine gene cluster , namely , P09341 , P19875 , P05231 , P20809 , P10145 , Q13007 and TGFbeta2 . In microarray studies , the radiation-induced up-regulated expression of some these genes encoding cytokine/chemokine proteins was significantly decreased by SeM treatment . For P10145 , TGFbeta2 , P09341 and P19875 , these observations were validated by qPCR techniques . It is concluded that SeM can regulate ionizing radiation-induced gene expression and may serve as an effective countermeasure for some of the acute inflammatory/immune responses induced by low-dose HZE-particle radiation . 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 . DB00159 inhibits prostaglandin D2 generation by inhibiting cyclo-oxygenase-2 in cultured human mast cells . BACKGROUND : DB00159 ( EPA ) is catalysed by cyclo-oxygenase ( P36551 ) , as is arachidonic acid , and is a competitive inhibitor of arachidonate metabolism . OBJECTIVES : We examined the effect of EPA on prostaglandin ( PG ) D2 generation in the cultured human mast cells with IgE-anti-IgE challenge incubation . METHODS : Cultured human mast cells were incubated with EPA ( 1 micromol/L ) for 20 h , then challenged with anti-IgE incubation after treatment with IgE . At the same time , P36551 inhibitors were tested to identify P23219 and P35354 activity . PGD2 synthetic activity was also assayed in a cell-free homogenate of cultured mast cells with P36551 inhibitors and EPA . DB11320 in the culture medium and in cells was assayed with the HPLC-fluorescent method . PGD2 and PGD3 were assayed with gas chromatography-mass spectrometry and the stable isotope dilution method . RESULTS : Although EPA incubation did not affect histamine release by cultured human mast cells in response to IgE-anti-IgE challenge incubation , it did decrease PGD2 generation by inhibiting the P35354 pathway . In contrast , in the cell-free homogenate of cultured human mast cells , EPA inhibited both P23219 and P35354 activities . CONCLUSION : Pre-incubation with EPA primarily affects the P35354 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE-anti-IgE challenge incubation . These findings suggest that P23219 and P35354 have different substrate flow systems in mast cells . They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti-inflammatory substrate in human mast cells . 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 . Central opioid inhibition of neuroendocrine stress responses in pregnancy in the rat is induced by the neurosteroid allopregnanolone . The hypothalamus-pituitary-adrenal ( Q9Y251 ) axis is the major neuroendocrine stress response system . P06850 ( P06850 ) neurons in the parvocellular paraventricular nucleus ( pPVN ) play a key role in coordinating responses of this system to stressors . The cytokine interleukin-1beta ( IL-1beta ) , mimicking infection , robustly activates these P06850 neurons via a noradrenergic input arising from the nucleus tractus solitarii ( P30990 ) . In late pregnancy , Q9Y251 axis responses to stressors , including IL-1beta , are attenuated by a central opioid mechanism that auto-inhibits noradrenaline release in the PVN . Here we show that the neuroactive progesterone metabolite allopregnanolone induces these changes in Q9Y251 responsiveness to IL-1beta in pregnancy . In late pregnancy , inhibition of 5alpha-reductase ( an allopregnanolone-synthesizing enzyme ) with finasteride restored Q9Y251 axis responses ( rapidly increased pPVN P06850 mRNA expression , DB01285 , and corticosterone secretion ) to IL-1beta . Conversely , allopregnanolone reduced Q9Y251 responses in virgin rats . In late pregnancy , activity of the allopregnanolone-synthesizing enzymes ( 5alpha-reductase and 3alpha-hydroxysteroid dehydrogenase ) was increased in the hypothalamus as was mRNA expression in the P30990 and PVN . Naloxone , an opioid antagonist , restores Q9Y251 axis responses to IL-1beta in pregnancy but had no additional effect after finasteride , indicating a causal connection between allopregnanolone and the endogenous opioid mechanism . Indeed , allopregnanolone induced opioid inhibition over Q9Y251 responses to IL-1beta in virgin rats . Furthermore , in virgin rats , allopregnanolone treatment increased , whereas in pregnant rats finasteride decreased proenkephalin-A mRNA expression in the P30990 . Thus , in pregnancy , allopregnanolone induces opioid inhibition over Q9Y251 axis responses to immune challenge . This novel opioid-mediated mechanism of allopregnanolone action may alter regulation of other brain systems in pregnancy . Calcimimetic and calcilytic drugs for treating bone and mineral-related disorders . The calcium-sensing receptor ( P41180 ) plays a pivotal role in regulating systemic Ca(2+) homeostasis and is a target for drugs designed to treat certain disorders of bone and mineral metabolism . Calcimimetics are agonists or positive allosteric modulators of the P41180 ; they inhibit parathyroid hormone ( PTH ) secretion and stimulate renal Ca(2+) excretion . The first calcimimetic drug is cinacalcet , a positive allosteric modulator of the P41180 that is approved for treating secondary hyperparathyroidism ( Q9HD23 ) in patients on renal replacement therapy and for some forms of primary Q9HD23 characterized by clinically significant hypercalcemia . DB01012 is also being investigated as a therapy for other hypercalcemic conditions and certain hypophosphatemic disorders . Calcilytics are P41180 inhibitors that stimulate the secretion of PTH and decrease renal excretion of Ca(2+) . Although calcilytics have failed thus far as anabolic therapies for osteoporosis , they are currently being evaluated as novel therapies for new indications involving hypocalcemia and/or hypercalciuria . Novel therapies for chronic lymphocytic leukemia . New therapies for chronic lymphocytic leukemia ( CLL ) are moving away from non-specific cytotoxic to more targeted approaches . The monoclonal antibody alemtuzumab induces responses in 33 % to 43 % of patients with relapsed or refractory disease , with 2-5 % CR . Side effects include infusional reactions as well as immunosuppressive effects . DB00073 has limited activity in relapsed refractory patients , but response rates are comparable to follicular non-Hodgkin 's lymphoma in untreated patients . Other antibodies in early phases of development include anti-CD23 [ IDEC-152 ] , anti- P20273 [ epratuzumab ] , Hu1D10 [ apolizumab ] , and anti- P33681 [ anti- P33681 , DB04901 ] . Other agents that are being studied include denileukin diftitox fusion protein ( Ontak ) , and bcl-2 antisense [ G3139 , Genasense ] . The mechanism of action of the new drugs and their role in CLL , as well as the emergence of new prognostic markers are discussed . P10997 -leptin coadministration stimulates central histaminergic signaling in rats . Combined amylin+leptin ( Q9BXJ7 + P41159 ) can reduce diet induced obesity and is very effective in combating P41159 resistance . The purpose of this study was to evaluate the effect of Q9BXJ7 + P41159 on central histaminergic signaling in lean and obese rats . Male rats were administered P41159 ( 300 μg/kg/d ) , Q9BXJ7 ( 100 μg/kg/d ) , Q9BXJ7 + P41159 or vehicle ( SAL , 0.9 % normal saline ) , via a subcutaneous mini-osmotic pump or single injection ( P41159 , 300 μg/kg and Q9BXJ7 , 100 μg/kg ) for acute studies . Q9BXJ7 + P41159 administration increased expression of histamine H1 receptor ( HIR ) and histidine decarboxylase ( HDC ) mRNA in the hypothalamus . Increased levels of P35367 were seen in arcuate ( Arc ) and ventromedial hypothalamus ( VMH ) as well as the area postrema ( APOS ) and nucleus of solitary tract ( P30990 ) following Q9BXJ7 + P41159 administration . APOS and P30990 also showed expression of immediate early gene c- P01100 in the hindbrain in Q9BXJ7 + P41159 -treated rats . We confirmed previous evidence indicating that Q9BXJ7 + P41159 increased P35610 -3 protein phosphorylation in Arc and VMH . Finally , by in vivo microdialysis , we observed an increase in methyl P42357 levels in the VMH of Q9BXJ7 , P41159 and Q9BXJ7 + P41159 -treated rats . Taken together , these observations are consistent with an important role that neuronal P42357 may play in mediating the potent effects of Q9BXJ7 + P41159 on food intake and body weight . 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 . Effect of P06850 on NO bioavailability , ROS production and antioxidant defense systems in endothelial EAhy926 cells . Local or ' Immune ' DB01285 -Releasing Hormone ( P06850 ) is secreted in peripheral tissues and plays a direct immunomodulatory role as an endocrine or paracrine mediator of inflammation . The present study was undertaken to determine whether P06850 affects the endothelial redox state . Accordingly , intracellular reactive oxygen species ( ROS ) content and peroxynitrite levels , endothelial nitric oxide synthase ( P29474 ) activity and nitric oxide ( NO ) levels as well as catalase activity , superoxide dismutase ( SOD ) activity and glutathione ( DB00143 ) levels were measured in the presence or absence of selective P06850 receptor-1 and P06850 receptor-2 inhibitors in endothelial EAhy926 cells exposed in vitro in 10(-7) M P06850 for 2 h . P06850 acting through both receptors induced a significant increase of ROS content ( p < 0.001 ) , catalase activity ( p < 0.001 ) and SOD activity ( p < 0.001 ) , accompanied by a simultaneous significant decrease of P29474 activity and NO levels ( p < 0.001 ) , as well as a significant increase in nitrotyrosine ( peroxynitrite ) levels ( p < 0.05 ) . The data indicate that P06850 may act as a regulator of pro-inflammatory mechanisms inducing adaptation of endothelial cell function to local stress . Baseline and P06850 -stimulated DB01285 and cortisol levels after administration of the peroxisome proliferator-activated receptor-gamma ligand , rosiglitazone , in Cushing 's disease . The ability of acute rosiglitazone administration in influencing DB01285 /cortisol secretion in basal conditions and after P06850 stimulation was studied in patients with Cushing 's disease . Ten patients ( 8 women and 2 men , aged 18-65 yr ) with Cushing 's disease were enrolled in the study : 6 of them had previously undergone unsuccessful surgery and 4 were untreated . Plasma DB01285 and serum cortisol levels were evaluated at serial time points for 3 h during saline infusion and after the administration of rosiglitazone ( 8 mg , po ) and for 1 h after the injection of P06850 ( 1 microg/kg iv ) given alone or 30 min following rosiglitazone administration . The 4 tests were performed in all subjects in randomized order on different days . No significant difference was observed between the pattern of hormone secretion during saline alone and after rosiglitazone , as evaluated by two-way analysis of variance ( Q9UNW9 ) . The integrated areas under the curves ( AUCs ) were also not significantly different ( DB01285 : 5683 +/- 1038 vs 6111 +/- 1007 pg/ml/180 min ; cortisol : 2333 +/- 267 vs 2902 +/- 486 microg/dl/180 min ) . In addition , there was no difference for DB01285 and cortisol responses to P06850 given either alone or after rosiglitazone , when evaluated as peak , increment or AUC ; the pattern of the responses analyzed by two-way Q9UNW9 was also similar . IN CONCLUSION : 1 ) the administration of a single dose of rosiglitazone did not decrease DB01285 /cortisol levels or blunt their response after P06850 injection ; 2 ) the activation of P37231 receptors by rosiglitazone seems unable to affect DB01285 and cortisol secretion , at least in acute conditions , in patients with DB01285 -secreting pituitary adenomas . 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 . Hypothalamic-pituitary adrenal response to cholecystokinin-B receptor agonism is resistant to cortisol feedback inhibition . Intravenous injection of the cholecystokinin ( CCK ) -B receptor agonist , pentagastrin , produces robust , dose-dependent release of adrenocorticotropin ( DB01285 ) and cortisol , supporting the hypothesis that CCK-B agonists pharmacologically activate the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . The mechanism of activation and its physiological relevance remain uncertain . Preliminary data suggest that the DB01285 response to pentagastrin may be differentiated from the response to exogenous corticotropin releasing hormone ( P06850 ) by its relative resistance to cortisol feedback inhibition . To more directly test the relationship between cortisol levels and DB01285 response to pentagastrin , this study examined responses to pentagastrin ( a ) during a peak ( 8 a.m. ) and a nadir ( 4 p.m. ) period of endogenous cortisol secretion and ( b ) when cortisol levels were artificially reduced to low levels by administration of metyrapone . DB01285 responses to pentagastrin were identical in the morning and afternoon , despite substantial differences in basal cortisol levels . Suppression of cortisol with metyrapone had little impact on DB01285 response to pentagastrin . These data support the hypothesis that P32239 mediated activation of the Q9Y251 axis is relatively resistant to cortisol feedback inhibition . This differentiates it from P06850 -mediated activation and raises the possibility that CCK could contribute to acute activation of the Q9Y251 axis even in the face of elevated basal cortisol levels , such as those seen in chronic stress or some psychiatric disorders . 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 .

MH_train_1494

MH_train_1494

MH_train_1494

interacts_with DB08816?

multiple_choice

DB00160 aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis . Plant peroxisomal glyoxylate aminotransferases play central roles within the photorespiratory pathway . Genes encoding glyoxylate aminotransferases have been isolated from several animals and microbes , but only recently have plant homologs been identified . Three Arabidopsis homologs of alanine (Ala):glyoxylate aminotransferase 2 ( Q9BYV1 ) contain a putative type 1 peroxisomal targeting signal ( PTS1 ) , but the metabolic significance of these Q9BYV1 homologs is unknown . P19440 and P36268 are Ala aminotransferase ( AlaAT ) homologs from Arabidopsis that represent another type of glyoxylate aminotransferase . These proteins are class I aminotransferases , each containing a putative PTS1 . P19440 and P36268 are members of a small family of AlaATs in Arabidopsis . When expressed as recombinant proteins in Escherichia coli , P19440 and P36268 displayed biochemical characteristics very similar to one another , and to the Arabidopsis protein purified from leaves . Four aminotransferase activities were specifically associated with P19440 and P36268 , using the substrate pairs glutamate ( DB00142 ):glyoxylate , Ala:glyoxylate , DB00142 :pyruvate , and Ala:2-oxoglutarate . P19440 and P36268 may have partially redundant functions ; transcripts of both genes were detected in many of the same tissues . Although DB00142 :glyoxylate aminotransferase ( P19440 ) activity has been observed in several locations in different plants and algae , including the cytoplasm and mitochondria , our subcellular fractionation data indicate that P19440 activity was exclusively peroxisomal in Arabidopsis . Thus , glyoxylate aminotransferase reactions in plant peroxisomes appear to be catalyzed by at least two distinct types of aminotransferases : an AGT1 homolog with serine:glyoxylate aminotransferase activity ( A.H. Liepman , L.J. Olsen [ 2001 ] Plant J 25 : 487-498 ) , and a pair of closely related , potentially redundant AlaAT homologs with P19440 activity . Partial trypsin digestion as an indicator of mis-folding of mutant alanine:glyoxylate aminotransferase and chaperone effects of specific ligands . Study of a spectrum of missense mutants . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme whose deficiency results in primary hyperoxaluria type 1 ( P78364 ) . More than 75 P78364 mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have previously demonstrated that many such mutants expressed by transcription/translation are subject to generalized degradation by the proteasome and a specific limited trimming by an endogenous DB00171 -independent protease activity . Here , we report the results of partial digestion using trypsin as a mimic for the endogenous non-proteasomal protease and the use of N-terminal protein sequencing to determine the sensitive site . Partial trypsin digestion also provided an indicator of proper folding of the mutant enzyme . For selected mutations the sensitivity to trypsin could be ameliorated by addition of pyridoxal phosphate or aminooxy acetic acid as specific pharmacological chaperones . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . 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 . 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 . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . 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 . Signaling responses to pulsatile gonadotropin-releasing hormone in LbetaT2 gonadotrope cells . The hypothalamic neuropeptide gonadotropin-releasing hormone ( DB00644 ) is secreted in a pulsatile fashion by hypothalamic neurons , and alterations in pulse frequency and amplitude differentially regulate gonadotropin synthesis and release . In this study , we investigated the kinetics of G(s) and G(q) signaling in response to continuous or pulsatile DB00644 using fluorescence resonance energy transfer reporters in live mouse LbetaT2 gonadotrope cells . DB02527 and protein kinase A-dependent reporters showed a rapid but transient increase in fluorescence resonance energy transfer signal with increasing doses of constant DB00644 , and in contrast diacylglycerol ( DAG ) and calcium reporters showed a rapid and sustained signal . Multiple pulses of DB00644 caused multiple pulses of DB02527 and protein kinase A activation without desensitization , but the DAG and calcium reporters were rapidly desensitized resulting in inhibition of calcium and DAG responses . At the transcriptional level , both a DB02527 -dependent DB02527 -response element reporter and a DAG/calcium-dependent AP-1 reporter showed a pulse frequency-dependent increase in luciferase activity . However , constant DB00644 stimulation gave very little DB02527 -response element activation but very strong AP-1 activation . Based on these data , we propose that both the P30968 -G(s) and G(q) pathways are responsive to pulses of DB00644 , but only the G(q) pathway is responsive to constant DB00644 . Furthermore , the G(q) pathway is subject to desensitization with multiple DB00644 pulses , but the G(s) pathway is not . DB00644 ( DB00644 ) receptor dynamics and gonadotrope responsiveness during and after continuous DB00644 stimulation . Gonadotrope responsiveness , serum and tissue levels of luteinizing hormone ( LH ) , and tissue concentration of gonadotropin-releasing hormone ( DB00644 ) receptors in ovariectomized rats were determined during and after continuous DB00644 stimulation . Intraperitoneal placement of DB00644 -containing osmotic minipumps for 96 h established a rate of DB00644 delivery ( 1 microgram/h ) that resulted in stable serum levels of DB00644 ( 500-700 pg/ml ) . Secretion of LH increased 8-fold within 6 h ; however , serum LH returned to pretreatment levels by 24 h , even with continued DB00644 stimulation . Tissue concentration of LH was depressed within 48 h of initiation of treatment but levels were restored by 96 h . Tissue levels of P30968 remained elevated during the first 6 h of treatment but were reduced by 60 % within 24 h and remained depressed for the duration of treatment . Gonadotrope responsiveness 48 h and 96 h after initiation of treatment was reduced by 50 % and 90 % , respectively . Removal of the DB00644 delivery vehicle resulted in rapid disappearance of DB00644 from serum . Dramatic reduction ( 75 % ) in circulating levels of LH , and a 2-fold increase in tissue levels of LH and in P30968 concentration were noted within 6 h of minipump removal . Although tissue concentration of P30968 returned to pretreatment levels within 48 h of minipump removal , both basal LH secretion and gonadotrope responsiveness remained depressed even 96 h after cessation of continuous DB00644 stimulation . These data indicate that DB00644 can " down regulate " its receptor , gonadotrope responsiveness is not obligatorily linked to receptor concentration , and desensitization that follows hyperstimulation represents effects directed at post-receptor loci . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . Selective Akt inhibitors synergize with tyrosine kinase inhibitors and effectively override stroma-associated cytoprotection of mutant P36888 -positive AML cells . OBJECTIVES : Tyrosine kinase inhibitor ( TKI ) -treated acute myeloid leukemia ( AML ) patients commonly show rapid and significant peripheral blood blast cell reduction , however a marginal decrease in bone marrow blasts . This suggests a protective environment and highlights the demand for a better understanding of stromal:leukemia cell communication . As a strategy to improve clinical efficacy , we searched for novel agents capable of potentiating the stroma-diminished effects of TKI treatment of mutant P36888 -expressing cells . METHODS : We designed a combinatorial high throughput drug screen using well-characterized kinase inhibitor-focused libraries to identify novel kinase inhibitors capable of overriding stromal-mediated resistance to TKIs , such as PKC412 and DB05213 . Standard liquid culture proliferation assays , cell cycle and apoptosis analysis , and immunoblotting were carried out with cell lines or primary AML to validate putative candidates from the screen and characterize the mechanism(s) underlying observed synergy . RESULTS AND CONCLUSIONS : Our study led to the observation of synergy between selective Akt inhibitors and P36888 inhibitors against mutant P36888 -positive AML in either the absence or presence of stroma . Our findings are consistent with evidence that Akt activation is characteristic of mutant P36888 -transformed cells , as well as observed residual Akt activity following P36888 inhibitor treatment . In conclusion , our study highlights the potential importance of Akt as a signaling factor in leukemia survival , and supports the use of the co-culture chemical screen to identify agents able to potentiate TKI anti-leukemia activity in a cytoprotective microenvironment . Q9BYW2 -α and survivin involved in the anti-apoptotic effect of DB02342 after global ischemia in rats . Survivin is an anti-apoptotic gene that decreases the apoptosis by depressing the expression of caspase-3 . Hypoxia-inducible factor-1-alpha ( Q16665 ) is a transcription factor specifically activated by hypoxia . 2-methoxyestradiol ( DB02342 ) is an estradiol derivative and a known Q16665 inhibitor . DB02342 decreased apoptosis by inhibiting Q16665 . The aim of the present study was to investigate if survivin is involved in the anti-apoptotic effect of DB02342 . Male adult rats were used to make the global ischemia ( GI ) model . Ten minutes after GI , DB02342 was injected intraperitoneally ( 16 mg/kg weight ) . Rats were killed at 6 hours , 12 hours , 24 hours , 48 hours , 96 hours , and 7 days . GI produced a marked increase in Q16665 expressions in the hippocampus at 6 hours and peaked at 48-96 hours . The expressions of survivin and caspase-3 were increased lightly in a similar time course . These molecular changes were accompanied by massive cell loss and apoptosis in the hippocampal regions . DB02342 treatment reduced the expression of Q16665 , increased survivin expression , and decreased the expression of caspase-3 . These results indicate that survivin and Q16665 were involved in the anti-apoptotic effect of DB02342 treated following GI . DB02342 may decrease the Q16665 expression , up-regulate the survivin expression , inhibit the expression of caspase-3 , and finally reduce apoptosis after GI . 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 . Anti- P01375 therapy . There are now five anti-tumour necrosis factor ( P01375 ) drugs licenced for use in rheumatoid arthritis . This chapter examines the similarities and differences between the drugs and looks for clues with regard to their rational prescribing . The major difference is between the monoclonal antibody-based drugs and the soluble receptor etanercept . DB00005 exhibits the best drug survival and is also associated with a lower risk of opportunistic infections , particularly tuberculosis . Immunogenicity should explain some of the differences between the different drugs but the lack of standardised assays has hindered this area of research . The optimal approach to the patient who has failed their first anti- P01375 remains unclear and awaits appropriate clinical trials . The safety profile of anti-TNFs has become clearer , largely through registry data . There is a small increase in serious and opportunistic infections but there does not appear to be a heightened cancer risk , and cardiovascular risk is probably reduced . Treatment of psoriatic arthritis with anti- P01375 agents : a systematic review and meta-analysis of efficacy , effectiveness and safety . We did a systematic review and meta-analysis on the efficacy and safety of the anti- P01375 drugs adalimumab , etanercept , DB06674 and infliximab used in psoriatic arthritis ( PsA ) adult treatment . Additionally , we present results of anti- P01375 use in real life settings . We searched Embase , Medline , Cochrane Central and LILACS , from inception to 11/08/2013 , for studies comparing anti-TNFs with each other or with controls . We included nine randomized controlled trials and six observational studies . ACR20 , ACR50 , PsARC and PASI75 responses were achieved by more users of anti- P01375 than control after up to 24 weeks of treatment . More participants who used etanercept and infliximab achieved ACR70 . After all patients originally randomized to anti- P01375 or placebo had used anti- P01375 for at least 24 weeks , we observed difference only with regard to ACR70 response . Radiographic end points were achieved by more patients in anti- P01375 group , and they seem to be time dependent-the longer patients use the drug the better the results . DB00005 and infliximab had worse results on application site reactions , but in general anti- P01375 drugs in the regimens studied were as safe as control/placebo . There seems to be no difference in efficacy and effectiveness among anti-TNFs , but superiority head-to-head studies are still needed . Meanwhile , other factors should be taken into account in the choice of medication , such as costs and patient convenience . P78364 -derived bivalent bibodies and trivalent tribodies bind differentially to shed and tumour cell-associated P15941 . Most adenocarcinomas express altered P15941 as a tumour-associated antigen . Due to suboptimal glycosylation in tumour-associated P15941 , the apomucin core is exposed , revealing new epitopes for antibody-directed immunotherapy . The human P78364 Fab binds specifically to this P15941 apomucin . We describe the engineering and functional characterization of bi- and trivalent recombinant antibody derivatives from the P78364 Fab . Bi- and tribodies were made using the disulfide-stabilized Fab fragment as a heterodimerization scaffold with P78364 single-chain variable fragments fused to either one or both Fab-chain C-termini . Immunoassays revealed 27- and 165-fold improved dissociation constants ( K(D) = 30 and 5 nM ) of the P78364 bi- and tribodies compared with the parental Fab ( K(D) = 820 nM ) . Unexpectedly , major differences were seen in the ability of the antibody constructs to bind shed and tumour cell-tethered P15941 . While the tribody did not discriminate between both P15941 forms , the bibody demonstrated preferential interaction with membrane-bound P15941 compared with shed P15941 . This preferential recognition of membrane-bound P15941 , along with the high serum stability of the bibody , its intermediate size and efficient internalization by P15941 (+) cells , makes the human P78364 -derived bibody a valuable candidate as a cancer-targeting therapeutic . DB08816 : positive , negative and misunderstood properties as a new antiplatelet agent . Dual antiplatelet therapy is essential for the management of acute coronary syndrome . In particular , combination therapy using aspirin with a platelet ADP ( i.e. Q9H244 ) receptor inhibitor , such as clopidogrel , prasugrel or , more recently , ticagrelor , has been recommended for patients with acute coronary syndrome . Pharmacological agents that reversibly inhibit platelet aggregation without metabolic activation in the liver are believed to reduce cardiovascular mortality compared with the current drug of choice for antiplatelet therapy , namely clopidogrel . These findings are based on a multicentre , double-blind , double-dummy , randomized controlled trial . Numerous factors are postulated to contribute to the improved survival of patients who take ticagrelor compared with those taking clopidogrel , including the risk of myocardial infarction , heart failure , arrhythmia and bleeding . In addition , clopidogrel may lead to a much higher incidence of infection . Although ticagrelor has recently been approved for use in the US and exhibits superiority over other antiplatelet agents , certain concerns remain regarding its use , including lung injury and dyspnoea , thus raising the issue of its true superiority over clopidogrel or prasugrel . Recent studies into ticagrelor report conflicting data , with certain aspects of its mechanisms of action still not fully understood . DB08816 has beneficial effects following its clinical application , such as achieving overall higher reductions in mortality compared with the use of clopidogrel and prasugrel . Harmful effects associated with the use of ticagrelor include a higher incidence of dyspnoea and major bleeding compared with clopidogrel . Biological characterization of DB05037 , a small-molecule inhibitor of cyclin-dependent kinases , in human tumor cell lines . P12004 -dependent kinases ( CDK ) , and their regulatory cyclin partners , play a central role in eukaryotic cell growth , division , and death . This key role in cell cycle progression , as well as their deregulation in several human cancers , makes them attractive therapeutic targets in oncology . A series of CDK inhibitors was developed using Astex 's fragment-based medicinal chemistry approach , linked to high-throughput X-ray crystallography . A compound from this series , designated DB05037 , is currently in early-phase clinical development . We describe here the biological characterization of DB05037 , a potent inhibitor of several CDK family members . DB05037 showed potent antiproliferative activity ( 40-940 nmol/L ) in a panel of human tumor cell lines , and the mechanism of action was shown here to be consistent with the inhibition of P06493 and P24941 in solid tumor cell lines . DB05037 caused cell cycle arrest followed by apoptosis in human tumor cells and inhibited tumor growth in human tumor xenograft models . Tumor regression was observed following twice daily dosing of DB05037 in the HCT116 and HT29 colon cancer xenograft models . We show that these biological effects are linked to inhibition of CDKs in vivo and that DB05037 induces tumor cell apoptosis in these xenograft models . DB05037 has an attractive biological profile for development as a clinical candidate , and the tolerability and efficacy in animal models compare favorably with other CDK inhibitors in clinical development . Studies described here formed the biological rationale for investigating the potential therapeutic benefit of DB05037 in cancer patients . Epstein-Barr virus can inhibit genotoxin-induced P55008 arrest downstream of p53 by preventing the inactivation of P24941 . Epstein-Barr virus ( EBV ) is involved in the pathogenesis of several B cell lymphoproliferations , but the precise contribution it makes to the aetiology of each remains unclear . In vitro , the virus has potent growth transforming activity and efficiently induces the continuous proliferation of normal human B cells . A comparison of EBV-infected primary B cells with an isogenic population induced to proliferate by P25942 -ligand ( P29965 ) and P05112 has revealed that EBV can override - by a novel mechanism - the p53/ P06400 -mediated P55008 checkpoint activated in normal B cells by a genotoxic stress . In cells responding to cisplatin , although p53 is stabilized and activated , EBV latent gene expression appears to inhibit the accumulation of newly synthesized P38936 ( P38936 /CIP1) and the downregulation of cyclin D2 that occur in the normal cells . Consequently , in the EBV-infected cells , P24941 remains active , hyperphosphorylation of P06400 is maintained and the replication of damaged DNA can occur . Under conditions of severe genomic stress , this absence of P38936 ( P38936 /CIP1) function can result in apoptosis ; however , when damage is less sustained , genomic instability may arise and this in turn could contribute to the development of a variety of EBV-associated B cell malignancies . Safety and efficacy of ticagrelor for neuroendovascular procedures . A single center initial experience . INTRODUCTION : Although platelet response testing is controversial , up to one-third of neuroendovascular patients are ' resistant ' to clopidogrel and are at risk for in stent thrombotic complications and may require alternative antiplatelet therapy . DB08816 is a new reversible ADP Q9H244 platelet receptor inhibitor with no known resistance . We describe the clinical experience with ticagrelor for neuroendovascular procedures as an alternative in clopidogrel Q9H244 platelet resistant patients . METHODS : We reviewed our cerebrovascular database for all patients who were non-responders to clopidogrel , defined as Q9H244 % inhibition < 30 % , despite repeat clopidogrel loading dose of at least 600 mg , and who were then administered ticagrelor . RESULTS : 18 patients were non-responders to clopidogrel ; 10 ( 56 % ) were men , eight ( 44 % ) were women , with a median age of 61 years ( range 38-84 ) . All patients received loading doses of at least 600 mg of clopidogrel and showed Q9H244 levels below 20 % prior to ticagrelor administration . Patients were loaded with 180 mg of ticagrelor , and all but one patient showed an initial Q9H244 response above 60 % . 11 patients underwent stenting , two underwent coiling , and five underwent treatment by pipeline embolization device . No patient experienced any adverse effects in the postoperative period related to the use of ticagrelor . CONCLUSIONS : DB08816 offers an effective alternative to clopidogrel non-responders . All of our patients showed immediate platelet inhibition after a loading dose of 180 mg of ticagrelor , with no adverse effects . The cost of medication , patient compliance ( twice a day doses ) , and reversible inhibition should be taken into consideration when using ticagrelor . 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 . Central role for hydrogen peroxide in P47900 ADP receptor-mediated cellular responses in vascular endothelium . ADP activates a family of cell surface receptors that modulate signaling pathways in a broad range of cells . ADP receptor antagonists are widely used to treat cardiovascular disease states . These studies identify a critical role for the stable reactive oxygen species hydrogen peroxide ( H2O2 ) in mediating cellular responses activated by the G protein-coupled P47900 receptor for ADP . We found that ADP-dependent phosphorylation of key endothelial signaling proteins -- including endothelial nitric oxide synthase , AMP-activated protein kinase , and the actin-binding P29966 protein -- was blocked by preincubation with PEG-catalase , which degrades H2O2 . ADP treatment promoted the H2O2-dependent phosphorylation of c-Abl , a nonreceptor tyrosine kinase that modulates the actin cytoskeleton . Cellular imaging experiments using fluorescence resonance energy transfer-based biosensors revealed that ADP-stimulated activation of the cytoskeleton-associated small GTPase Rac1 was independent of H2O2 . However , Rac1-dependent activation of AMP-activated protein kinase , the signaling phospholipid phosphatidylinositol- ( 4 , 5 ) -bisphosphate , and the c-Abl-interacting protein CrkII are mediated by H2O2 . We transfected endothelial cells with differentially targeted HyPer2 H2O2 biosensors and found that ADP promoted a marked increase in H2O2 levels in the cytosol and caveolae , and a smaller increase in mitochondria . We performed a screen for P47900 receptor-mediated receptor tyrosine kinase transactivation and discovered that ADP transactivates P36888 ( Flt3 ) , a receptor tyrosine kinase expressed in these cells . Our observation that P47900 receptor-mediated responses involve Flt3 transactivation may identify a unique mechanism whereby cancer chemotherapy with receptor tyrosine kinase inhibitors promotes vascular dysfunction . Taken together , these findings establish a critical role for endogenous H2O2 in control of ADP-mediated signaling responses in the vascular wall . DB05387 ( AEterna ) . AEterna is developing DB05387 , an angiogenesis inhibitor derived from the ultrafiltration of liquid shark cartilage , with matrix metalloprotease ( MMP ) -2 , P14780 and vascular endothelial growth factor inhibitory properties , for the potential treatment of non-small-cell lung cancer . Hypoxia inducible factor 1 alpha regulates matrigel-induced endovascular differentiation under normoxia in a human extravillous trophoblast cell line . Extravillous trophoblast ( EVT ) cells mimic endothelial cells during angiogenesis , inducing remodeling of the spiral arteries that increases blood flow toward the intravillous space . We have previously shown that signals involving the vascular endothelial growth factor ( P15692 ) axis are essential for endovascular differentiation through integrin signaling from the extracellular matrix : This was accomplished with use of the human EVT cell line TCL1 , which shows tube formation that specifically recalls morphological changes in endothelial cells . To investigate endovascular differentiation in EVT further , we investigated the role of hypoxia inducible factor (HIF)1A , a subunit of Q9BYW2 transcription factor that regulates not only adaptive responses to hypoxia , but also many cellular functions under normoxia , which was up-regulated in DNA microarray analysis during matrigel-induced endovascular differentiation under normoxia . Q16665 induces P15692 and P06756 / P05106 aggregation , actions known to be important for cellular survival and endovascular differentiation in EVT . Inhibition of Q16665 up-regulation using siRNA introduction or chemical inhibition suppressed hypoxia-responsive element transcriptional activity , P15692 induction , P06756 / P05106 aggregation accompanied by the inhibition of tube formation in TCL1 cells . These results suggest that Q16665 has a crucial role in regulating EVT behavior including matrigel-induced endovascular differentiation under normoxia . Relationship between soluble P29965 and gamma-glutamyltransferase concentrations in non-drinking , young Type 1 diabetic individuals . AIMS : To assess the association between circulating levels of soluble P29965 ( sCD40L ) , an emerging cardiovascular risk factor , and gamma-glutamyltransferase ( P19440 ) activity concentrations in Type 1 diabetic subjects . METHODS : Plasma concentrations of sCD40L and P19440 activity , a marker of liver dysfunction , were measured in 54 non-smoking , non-drinking , young Type 1 diabetic patients , who were free of diagnosed cardiovascular disease . RESULTS : When participants were grouped according to tertiles of P19440 , plasma sCD40L concentrations steadily increased across P19440 tertiles ( P = 0.007 for trend ) . Similarly , plasma sCD40L concentrations were positively correlated with plasma P19440 levels in the whole group of participants ( r = 0.532 ; P < 0.0001 ) . In multivariate linear regression analysis , plasma P19440 activity levels were positively associated with sCD40L ( standardized beta coefficient = 0.342 ; P = 0.027 ) independently of age , gender , diabetes duration , glycated haemoglobin , total cholesterol and systolic blood pressure . Further adjustment for the presence of diabetic retinopathy and microalbuminuria did not appreciably attenuate this association . CONCLUSIONS : Our findings suggest that there is a strong , graded , relationship between plasma P19440 activity and sCD40L concentrations in non-smoking , non-drinking , young Type 1 diabetic individuals . This association appears to be independent of numerous confounding factors . Further studies are required to confirm the reproducibility of these results . P29474 uncoupling in cardiovascular diseases -- the role of oxidative stress and inflammation . Many cardiovascular diseases and drug-induced complications are associated with - or even based on - an imbalance between the formation of reactive oxygen and nitrogen species ( RONS ) and antioxidant enzymes catalyzing the break-down of these harmful oxidants . According to the " kindling radical " hypothesis , the formation of RONS may trigger in certain conditions the activation of additional sources of RONS . According to recent reports , vascular dysfunction in general and cardiovascular complications such as hypertension , atherosclerosis and coronary artery diseases may be connected to inflammatory processes . The present review is focusing on the uncoupling of endothelial nitric oxide synthase ( P29474 ) by different mechanisms involving so-called " redox switches " . The oxidative depletion of tetrahydrobiopterin ( BH4 ) , oxidative disruption of the dimeric P29474 complex , S-glutathionylation and adverse phosphorylation as well as RONS-triggered increases in levels of asymmetric dimethylarginine ( DB01686 ) will be discussed . But also new concepts of P29474 uncoupling and state of the art detection of this process will be described . Another part of this review article will address pharmaceutical interventions preventing or reversing P29474 uncoupling and thereby normalize vascular function in a given disease setting . We finally turn our attention to the inflammatory mechanisms that are also involved in the development of endothelial dysfunction and cardiovascular disease . Inflammatory cell and cytokine profiles as well as their interactions , which are among the kindling mechanisms for the development of vascular dysfunction will be discussed on the basis of the current literature . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . 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 . Purinergic receptors are part of a functional signaling system for proliferation and differentiation of human epidermal keratinocytes . We investigated the expression of Q93086 , Q99572 , P47900 and P41231 receptor subtypes in normal human epidermis and in relation to markers of proliferation ( P12004 and Ki-67 ) , keratinocyte differentiation ( cytokeratin P13645 and involucrin ) and markers of apoptosis ( TUNEL and anticaspase-3 ) . Using immunohistochemistry , we showed that each of the four receptors was expressed in a spatially distinct zone of the epidermis , suggesting different functional roles for these receptors . Functional studies were performed on primary cultures of human keratinocytes and on explanted rat skin , where different P2 receptor subtype agonists and antagonists were applied to cultured keratinocytes or injected subcutaneously into the skin , respectively . An increase in cell number was caused by low doses of the nonspecific P2 receptor agonist DB00171 , the P41231 receptor agonist UTP ( p < 0.001 ) , and the P47900 receptor agonist 2MeSADP ( p < 0.05 ) . There was a significant decrease in cell number as a result of treatment with the Q93086 receptor agonist ATPgammaS ( p < 0.001 ) and the Q99572 receptor agonist BzATP ( p < 0.001 ) . Suramin caused a significant block in the effect of 100 microm DB00171 ( p < 0.01 ) and 1000 microm DB00171 ( p < 0.001 ) on cell number . These results imply that different purinergic receptors have different functional roles in the human epidermis with P47900 and P41231 receptors controlling proliferation , while Q93086 and Q99572 receptors control early differentiation , terminal differentiation and death of keratinocytes , respectively . DB08816 yields consistent dose-dependent inhibition of ADP-induced platelet aggregation in patients with atherosclerotic disease regardless of genotypic variations in Q9H244 , P47900 , and P05106 . The platelet P2Y(12) receptor is the target of clopidogrel therapy , which has been shown to reduce thromboembolic complications of atherosclerotic disease but has limitations in terms of variability of response and irreversibility of effect . This receptor is also a target for ticagrelor ( AZD6140 ) , the first reversibly binding oral P2Y(12) receptor antagonist that does not require metabolic activation and yields more consistent inhibition of platelet aggregation than clopidogrel therapy . Single nucleotide polymorphisms ( SNPs ) have been described in the gene for this receptor ( Q9H244 ) , some of which have been associated with variability in platelet reactivity . SNPs in P47900 and P05106 have also been reported by some groups to affect platelet reactivity to adenosine diphosphate ( ADP ) . We assessed whether SNPs in these genes influenced the pharmacodynamic response to ticagrelor in patients enrolled in both the DISPERSE study ( stable atherosclerotic disease ) and the DISPERSE2 study ( non-ST-segment elevation acute coronary syndromes ) . Platelet aggregation data ( at baseline and 4 weeks ) and DNA samples from clopidogrel-naive Caucasian patients treated with ticagrelor were available for 151 patients . Seventy four SNPs within three genes were genotyped . After adjustment for multiple comparisons , none of these SNPs were found to significantly influence inhibition of ADP-induced platelet aggregation by ticagrelor . Mechanism of purinergic activation of endothelial nitric oxide synthase in endothelial cells . BACKGROUND : Decreased endothelial nitric oxide ( NO ) synthase ( P29474 ) activity and NO production are critical contributors to the endothelial dysfunction and vascular complications observed in many diseases , including diabetes mellitus . Extracellular nucleotides activate P29474 and increase NO generation ; however , the mechanism of this observation is not fully clarified . METHODS AND RESULTS : To elucidate the signaling pathway(s) leading to nucleotide-mediated P29474 phosphorylation at DB00133 -1177 , human umbilical vein endothelial cells were treated with several nucleotides , including DB00171 , UTP , and ADP , in the presence or absence of selective inhibitors . These experiments identified P47900 , P41231 , and possibly P51582 as the purinergic receptors involved in P29474 phosphorylation and demonstrated that this process was adenosine independent . Nucleotide-induced P29474 phosphorylation and activity were inhibited by BAPTA-AM ( an intracellular free calcium chelator ) , rottlerin ( a protein kinase Cdelta inhibitor ) , and protein kinase Cdelta siRNA . In contrast , blockade of AMP-activated protein kinase , calcium/calmodulin-dependent kinase II , calcium/calmodulin-dependent kinase kinase , serine/threonine protein kinase B , protein kinase A , extracellular signal-regulated kinase 1/2 , and p38 mitogen-activated protein kinase did not affect nucleotide-mediated P29474 phosphorylation . CONCLUSIONS : The present study indicates that extracellular nucleotide-mediated P29474 phosphorylation is calcium and protein kinase Cdelta dependent . This newly identified signaling pathway opens new therapeutic avenues for the treatment of endothelial dysfunction . Direct oral anticoagulants in acute coronary syndrome . Patients with acute coronary syndromes ( ACS ) require a specific antithrombotic therapy in the immediate and the post ACS phase . The current antithrombotic therapy in the acute phase of an ACS combines antiplatelet and anticoagulant drugs in order to reduce ischemic cardiovascular events . In the post ACS phase , dual antiplatelet therapy ( DAPT ; aspirin and a Q9H244 receptor antagonist ) is the current mainstay of antithrombotic treatment and is recommended in the guidelines of the major North American and European clinical cardiology associations ( DB00551 , ACC , and ESC ) . Recently , the addition of rivaroxaban , a low dose oral direct factor Xa inhibitor ( 2.5 mg twice daily ) , to DAPT ( aspirin plus second-generation Q9H244 inhibitor ) showed a significant reduction of cardiovascular and overall mortality in the major phase III clinical trial ATLAS ACS 2 TIMI 51 . This led to the approval of low-dose rivaroxaban in addition to aspirin and clopidogrel by the European Medicines Agency ( P15941 ) in 2013 . Other direct oral anticoagulants ( apixaban , dabigatran etexilate ) have also been assessed in phase II ( dabigatran etexilate ) and phase III ( apixaban ) post ACS clinical trials . In the studied dosing regimens , these drugs failed to show a net clinical benefit in addition to dual antiplatelet therapy . The major clinical phase II and III post ACS studies of direct oral anticoagulants are summarized and discussed in this article along with the concept of long-term anticoagulation for the secondary prevention of ischemic events after ACS and implications for the future of antithrombotic therapy in the current era of third-generation Q9H244 receptor inhibitors ( Prasugrel and DB08816 ) . 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 . Piceatannol is more effective than resveratrol in restoring endothelial cell dimethylarginine dimethylaminohydrolase expression and activity after high-glucose oxidative stress . DB09341 -induced oxidative stress is involved in endothelial dysfunction . Dimethylarginine dimethylaminohydrolase ( DDAH ) and arginase are regulators of the endothelial NO synthase ( P29474 ) . This study aimed to compare the effect of two polyphenolic antioxidants , resveratrol and piceatannol , on DDAH and arginase pathways in bovine aortic endothelial cells under 25 mM glucose for 24 h . DDAH activity and expression were decreased in these cells as compared to control cells , whereas arginase activity was unchanged . DDAH inhibition led to intracellular accumulation of asymmetric dimethylarginine ( DB01686 ) , a natural inhibitor of P29474 . Under these conditions , cell pre-treatment with resveratrol ( 0.1-10 μM ) restored basal DDAH activity and DB01686 level with a dose-dependent effect . Piceatannol acted as resveratrol on DDAH pathway but at 10-fold lower concentrations . DB02709 and piceatannol restored DDAH activity even in the presence of splitomicin , a specific inhibitor of Sirtuin 1 . These results suggest potential therapeutic intervention targeting resveratrol or piceatannol administration to improve endothelial dysfunction .

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interacts_with DB01656?

multiple_choice

Ventilatory strategies in obstructive lung disease . Chronic obstructive pulmonary disease ( P48444 ) is characterized by expiratory flow limitation ( EFL ) due to progressive airflow obstruction . The various mechanisms that cause EFL are central to understanding the physiopathology of P48444 . At the end of expiration , dynamic inflation may occur due to incomplete emptying the lungs . This " extra " volume increases the alveolar pressure at the end of the expiration , resulting in auto-positive end-expiratory pressure ( PEEP ) or PEEPi . Acute exacerbations of P48444 may result in increased airway resistance and inspiratory effort , further leading to dynamic hyperinflation . P48444 exacerbations may be triggered by environmental exposures , infections ( viral and bacterial ) , or bronchial inflammation , and may result in worsening respiratory failure requiring mechanical ventilation ( MV ) . Acute exacerbations of P48444 need to be distinguished from other events such as cardiac failure or pulmonary emboli . Strategies to treat acute respiratory failure ( Q8N726 ) in P48444 patients include noninvasive ventilation ( NIV ) , pressure support ventilation , and tracheal intubation with MV . In this review , we discuss invasive and noninvasive techniques to address Q8N726 in this patient population . When invasive MV is used , settings should be adjusted in a way that minimizes hyperinflation , while providing reasonable gas exchange , respiratory muscle rest , and proper patient-ventilator interaction . Further , weaning from MV may be difficult in these patients , and factors amenable to pharmacological correction ( such as increased bronchial resistance , tracheobronchial infections , and heart failure ) are to be systematically searched and treated . In selected patients , early use of NIV may hasten the process of weaning from MV and improve outcomes . Targeted inhibition of P15692 receptor 2 : an update on ramucirumab . INTRODUCTION : DB05578 ( IMC-1121B ) is a fully humanized IgG1 monoclonal antibody , targeting the extracellular domain of P15692 receptor 2 ( P35968 ) . Numerous Phase I - II trials in various malignancies have shown promising clinical antitumor efficacy and tolerability . Most recently , the large Phase III REGARD trial evaluated ramucirumab in patients with refractory metastatic gastric cancer . Patients receiving ramucirumab experienced a median overall survival of 5.2 months compared to 3.8 months on placebo . AREAS COVERED : The purpose of this article is to review the preclinical motivation for P35968 -targeted therapies and survey recent data from clinical trials involving ramucirumab , as well as highlight ongoing studies . EXPERT OPINION : Rational multi-target approaches to angiogenesis are needed to overcome resistance mechanisms . Predictive angiogenic biomarkers are also needed to optimize patient selection for novel anti-angiogenic agents . P11926 over-expression stimulates mitogen-activated protein kinase and anchorage-independent growth of human breast epithelial cells . In these experiments we tested the hypothesis that constitutive activation of polyamine(PA) biosynthesis may contribute to mammary carcinogenesis . Spontaneously immortalized normal human MCF-10A breast epithelial cells were infected with the retroviral vector pLOSN containing a cDNA which codes for a truncated and more stable ornithine decarboxylase ( ODC ) , the rate-limiting enzyme in PA synthesis . Upon chronic selective pressure with alpha-difluoromethyl-ornithine ( DB06243 ) ( an irreversible inhibitor of ODC ) , infected MCF-10A cells exhibited an approximately 250-fold increase in ODC activity , which persisted despite discontinuation of DB06243 . ODC-over-expressing MCF-10A cells showed a modest decrease in S-adenosylmethionine decarboxylase and an increase in spermidine/spermineN1-acetyltransferase . Analysis of cellular PA profile revealed a selective accumulation of putrescine without alterations in spermidine and spermine contents . Lesser degrees of increased ODC activity were obtained reproducibly by re-exposing the cells to incremental small doses of DB06243 . We observed a bell-shaped dose-related positive effect of ODC activity on clonogenicity in soft agar of MCF-10A cells . Since anchorage-dependent growth was actually reduced , such positive influence on this feature of transformation was not a non-specific consequence of a growth advantage provided by ODC over-expression . In addition , we observed a close parallelism between the dose-dependent effects of ODC expression on clonogenicity and activity of the P28482 kinase , a central element of the MAPK cascade . Our data demonstrate an interaction between PA and the MAPK signalling pathway and suggest that the latter may be involved in ODC-induced transformation of mammary epithelial cells . 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 . The role of heat shock proteins in Amyotrophic Lateral Sclerosis : The therapeutic potential of DB05025 . DB05025 is a hydroxylamine derivative , a group of compounds which have unique properties as co-inducers of heat shock protein expression , but only under conditions of cellular stress . DB05025 has been found to be neuroprotective in a number of neurodegenerative disease models , including Amyotrophic Lateral Sclerosis ( P35858 ) , and in mutant Superoxide Dismutase 1 ( P00441 ) mice that model P35858 , DB05025 rescues motor neurons , improves neuromuscular function and extends lifespan . The therapeutic potential of DB05025 is currently under investigation in a Phase II clinical trial for P35858 patients with P00441 mutations . In this review we summarize the evidence for the neuroprotective effects of enhanced heat shock protein expression by DB05025 and other inducers of the Heat Shock Response . P35858 is a complex , multifactorial disease affecting a number of cell types and intracellular pathways . Cells and pathways affected by P35858 pathology and which may be targeted by a heat shock protein-based therapy are also discussed in this review . For example , protein aggregation is a characteristic pathological feature of neurodegenerative diseases including P35858 . Enhanced heat shock protein expression not only affects protein aggregation directly , but can also lead to more effective clearance of protein aggregates via the unfolded protein response , the proteasome-ubiquitin system or by autophagy . However , compounds such as DB05025 have effects beyond targeting protein mis-handling and can also affect additional pathological mechanisms such as oxidative stress . Therefore , by targeting multiple pathological mechanisms , compounds such as DB05025 may be particularly effective in the development of a disease-modifying therapy for P35858 and other neurodegenerative disorders . Amyotrophic lateral sclerosis ( P35858 ) , a novel rare cause of elevated plasma troponin T levels . In this article , we report on a patient with chronic and modestly elevated plasma troponin T ( TnT ) levels and frequent hospitalizations following the first admission until his death one year later . The patient was initially admitted for dyspnea and discharged from hospital with a diagnosis of non-ST elevation acute myocardial infarction ( AMI ) . Coronary angiography and echocardiography were normal , but the patient received the ( false ) diagnosis of AMI at two further admissions , based purely on elevated TnT . Shortly thereafter , severe respiratory failure with restrictive-type spirometry pattern became the predominant clinical symptom , with constantly elevated TnT levels at frequent re-admissions . Due to inconsistent follow-up by primarily junior and non-specialist staff at a number of different wards , pulmonary function tests and previous smoking history were mis-interpreted as typical of chronic obstructive pulmonary disease ( P48444 ) . The patient received standard P48444 treatment without any improvement . After a year of gradually worsening respiratory failure and repeated hospitalizations , thorough assessment by a pulmonologist and neurologist established the final diagnosis of amyotrophic lateral sclerosis ( P35858 ) . The patient died shortly thereafter . While progressive respiratory failure is well-known to determine morbidity and mortality in patients with P35858 , chronically elevated TnT levels in the absence of coronary artery disease have , to our best knowledge , not been described so far . We suggest that chronic myocardial hypoxia due to P35858 -related hypoxic respiratory failure was the most likely underlying etiology for the elevated TnT levels seen here but other mechanism such as immune-mediated myocardial injury can not be excluded . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . The 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 . DB05332 as a treatment for immune thrombocytopenia : a review . " Immune thrombocytopenia " ( ITP ) is an autoimmune disorder that leads to peripheral destruction , as well as a decreased production of platelets . ITP most commonly presents as mild mucocutaneous bleeding . Though it is rare , the leading cause of mortality in persons with ITP is intracranial hemorrhage and those that do not respond to therapy are at increased risk . Our understanding of the pathophysiology of ITP has evolved immensely , especially over the last 60 years . The discovery of the platelet-production stimulator , thrombopoietin ( P07202 ) , lent clarity to an earlier hypothesis that inhibition of platelet production at the level of the megakaryocyte , at least in part , accounts for thrombocytopenia in adults with ITP . This facilitated the development of P07202 -based therapies to treat ITP . P40238 agonists are one of the most recent treatments to enter the landscape . Original production of a recombinant human P07202 was halted after clinical trials revealed the untoward effect of autoantibodies to the recombinant human P07202 with cross-reactivity to endogenous P07202 . Next-step development focused on stimulation of the P07202 receptor with fewer immunogenic agents . Currently , two such thrombopoietin receptor agonists , romiplostim and eltrombopag , are licensed in the USA to treat thrombocytopenia in adults with persistent or chronic ITP . Ongoing research will assess their efficacy in other immune-mediated and nonimmune-mediated primary and secondary thrombocytopenias . Increased miR-21 expression during human monocyte differentiation into DCs . Differentiation of monocytes into dendritic cells ( DCs ) is characterised by marked changes in gene expression . The role of microRNAs ( miRNAs ) , a new class of small endogenous non-coding regulatory RNAs , in this process is still unclear . We identified miR-223 , miR-16 , miR-191 , miR-24 , let-7b , and miR-21 as differentially expressed between monocytes and monocyte derived DCs . We evaluated the expression levels of computationally predicted target genes of miR-21 in human monocytes following stimulation with GM- P04141 and P05112 . Moreover , transfection of monocytes with synthetic miR-21 inhibited the expression of a set of genes that were also repressed during monocyte differentiation to DCs in response to GM- P04141 and P05112 . Among these , we identified genes that are involved in cell cycle , apoptosis and differentiation such as Q07343 , Q53EL6 , P04083 , Q9NXR8 , Q8N3U4 , Q15582 , P80511 , LAT2 and P48552 . Collectively , the present study highlights the involvement of miRNAs , particularly miR-21 in monocyte differentiation to DCs and identifies potential miR-21 target genes . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . P05305 downregulates angiotensin-converting enzyme-2 expression in human bronchial epithelial cells . BACKGROUND/AIMS : Both endothelin-1 ( ET-1 ) and the renin-angiotensin system ( DB01367 ) are implicated in the pathogenesis and progression of chronic obstructive pulmonary disease ( P48444 ) . In the present study , we explored the interaction between ET-1 and the DB01367 by examining the effect of ET-1 on angiotensin-converting enzyme-2 ( Q9BYF1 ) expression and activity in human bronchial epithelial cells ( HBEpCs ) . METHODS : HBEpCs were treated with ET-1 ( 1 , 10 , 20 , 40 or 50 nmol/l ) for 6 , 12 , 18 , 24 or 30 h with or without the transcription inhibitor actinomycin D , endothelin A ( P25101 ) receptor blocker BQ123 , endothelin B receptor blocker BQ788 , or different kinase inhibitors . RESULTS : ET-1 decreased the Q9BYF1 mRNA level in a dose- and time-dependent manner within 24 h , which led to dose-dependent downregulation of the Q9BYF1 promoter activity , protein level and the cell membrane Q9BYF1 activity . DB00970 ( 1 mg/ml ) , BQ123 ( 1 μmol/l ) , and the p38 mitogen-activated protein kinase ( MAPK ) siRNA and inhibitor PD169316 ( 25 μmol/l ) completely abolished the effect of ET-1 on Q9BYF1 expression in HBEpCs . CONCLUSION : ET-1 downregulates Q9BYF1 expression and activity at the transcription level in HBEpCs via the P25101 receptor by a p38 MAPK-dependent mechanism . This is the first evidence of crosstalk between the ET-1/ P25101 axis and the DB01367 in regard to the pathogenesis and progression of P48444 . 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 . DuOx2 Promoter Regulation by Hormones , Transcriptional Factors and the Coactivator TAZ . The production of H2O2 , which is essential to thyroid hormone synthesis , involves two NADPH oxidases : dual oxidases 1 and 2 ( DuOx1 and DuOx2 ) . A functional study with human DuOx genes and their 5'-flanking regions showed that DuOx1 and -2 promoters are different from thyroid-specific gene promoters . Furthermore , their transcriptional activities are not restricted to thyroid cells . While regulation of Tg ( thyroglobulin ) and P07202 ( thyroperoxidase ) expression have been extensively studied , DuOx2 promoter regulation by hormones and transcriptional factors need to be more explored . Herein we investigated the role of DB00024 , insulin and insulin-like growth factor 1 ( DB01277 ) , as well as the DB02527 effect on DuOx2 promoter ( ptx41 ) activity in transfected rat thyroid cell lines ( PCCL3 ) . We also assessed DuOx2 promoter activity in the presence of transcriptional factors crucial to thyroid development such as Q15669 -1 ( thyroid transcription factor 1 ) , Q06710 , CREB , Q9Y2W7 , Nkx2.5 and the coactivator TAZ in HeLa and P29320 293T-transfected cells . Our results show that DB00024 and forskolin , which increase DB02527 in thyroid cells , stimulated DuOx2 promoter activity . DB01277 led to pronounced stimulation , while insulin induction was not statistically different from DuOx2 promoter basal activity . All transcriptional factors selected for this work and coactivator TAZ , except Q9Y2W7 , stimulated DuOx2 promoter activity . Moreover , Nkx2.5 and TAZ synergistically increased DuOx2 promoter activity . In conclusion , we show that DuOx2 expression is regulated by hormones and transcription factors involved in thyroid organogenesis and carcinogenesis , reinforcing the importance of the control of H2O2 generation in the thyroid . 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 . DB00588 and Salmeterol combination induces O14543 expression in airway epithelial cells . DB00588 ( FP ) and Salmeterol ( SAL ) are commonly used in combination therapy for patients with Chronic obstructive pulmonary disease ( P48444 ) . Clinical studies show that FP/SAL used in combination therapy was found to inhibit airway inflammation in P48444 patients . However , the mechanisms associated with FP/SAL induced anti-inflammatory effects were not clear . We have evaluated the effect of FP/SAL and tobacco smoke ( TS ) on O14543 and interleukine-6 expression in bronchial airway epithelial cells ( BAEpCs ) . Human BAEpCs were exposed to TS and subsequently treated with FP or SAL alone or in combinations in the presence and absence of mitogen activated protein kinase ( MAPK ) inhibitors for either Erk1/Erk2 , or p38 or P19957 kinase . In BAEpCs , TS induced P05231 expression via P27361 / P28482 MAPK pathway and FP/SAL inhibited TS mediated P05231 expression . TS down regulated the O14543 expression via activation of Erk1/Erk2 , and p38 MAPK signaling . When TS exposed BAEpCs were treated with FP/SAL O14543 expression was restored . FP/SAL combinations induced significantly higher expression of O14543 in BAEpCs when compared to individual drug . Pretreatment with Ly294002 a P19957 MAPK inhibitor significantly attenuated FP/SAL induced O14543 expression in BAEpCs . Furthermore , FP/SAL blunted TS induced phosphorylation of Erk1/Erk2 and p38 MAPK in BAEpCs . Our study suggests that TS inhibits O14543 , combination of FP/SAL has a profound synergistic effect on O14543 induction in BAEpCs and it is dependent on P19957 kinase signaling pathway . O14543 may represent a potential biomarker for understanding the efficacy and a novel anti-inflammatory mechanism of FP/SAL combination therapy in the treatment of P48444 . DB00142 decarboxylase in cerebrospinal fluid in infancy and childhood . Part I . DB00142 decarboxylase activity in cerebrospinal fluid of normal infants and children . DB00142 decarboxylase ( Q99259 ) activity in the cerebrospinal fluid ( P04141 ) of normal infants ( n:14 ) and children ( n:28 ) was determined by measuring the amount of 14CO2 released from L-[1-14C]-glutamic acid . The mean Q99259 activity in P04141 of infants and children was 5.2 +/- 2.5 pmol CO2 formed/hr/ml . Dividing these subjects into 4 groups according to age , Q99259 activities in P04141 were 5.4 +/- 1.6 pmol CO2 formed/hr/ml in neonates ( 0-1 m ) , 3.6 +/- 1.6 pmol CO2 formed/hr/ml in infants ( 2-12 m ) , 3.9 +/- 1.1 pmol CO2 formed/hr/ml in young children ( 2-6 yr ) and 7.1 +/- 2.3 pmol CO2 formed/hr/ml in school children ( 7-16 yr ) , respectively . In neonates and school children , Q99259 activities were significantly higher ( p less than 0.001 ) than those in the other age groups . In infants under 6 months of age , a significantly negative correlation between Q99259 activity in P04141 and their ages was recognized ( r = -0.52 , p less than 0.001 ) . In infants and children ranging from 6 months to 16 years of age , a significantly positive correlation between Q99259 activity in P04141 and their ages was found ( r = 0.67 , p less than 0.001 ) . These data suggest that high Q99259 activity in neonates may be due to hypoxia at birth and the activity gradually increases from 6 months to 15 years of age . P62937 is required for Q9C035 {alpha}-mediated resistance to HIV-1 in Old World monkey cells . The peptidyl-prolyl isomerase cyclophilin A ( CypA ) embraces an exposed , proline-rich loop on HIV-1 capsid ( CA ) and renders reverse transcription complexes resistant to an antiviral activity in human cells . A CypA fusion with Q9C035 that is unique to New World owl monkeys also targets HIV-1 CA , but this interaction potently inhibits infection . A similar block to HIV-1 infection in Old World monkeys is attributable to the alpha isoform of the Q9C035 orthologue in these species . To determine whether HIV-1 restriction by Old World monkey TRIM5alpha is modulated by the CA-CypA interaction , RNA interference was used to disrupt CypA in cells from African green monkeys and rhesus macaques . HIV-1 infectivity increased in response to CypA knock-down to the same extent that it increased in response to Q9C035 knock-down . CypA knock-down eliminated the HIV-1 stimulatory effect of cyclosporin A ( DB00091 ) , a competitive inhibitor of the CypA-CA interaction , or of CA mutants that block binding to CypA but caused no change in titer of retroviruses that do n't interact with CypA . Simultaneous knock-down of both CypA and Q9C035 caused minimal additional increase in titer , suggesting that CypA inhibits HIV-1 replication in these cells because it is required for CA recognition by TRIM5alpha . Finally , DB00091 increased HIV-1 titer in otherwise nonrestrictive feline cells but only after these cells were transduced with Old World monkey TRIM5alpha . Thus , CypA is required for HIV-1 restriction by Old World monkey orthologues of TRIM5alpha . Anti-inflammatory effect of transduced PEP-1-cyclophilin A in Raw264.7 cells and 12-O-tetradecanoylphorbol-13-acetate-induced mice . AIMS : P62937 ( CypA ) is an immunophilin that acts as a receptor for the immunosuppressant drug cyclosporine A ( DB00091 ) . CypA has emerged as a potential drug target for several inflammatory diseases , although the details of its mechanism are unclear . We examined the protective effects of CypA on inflammation in Raw 264.7 cells and animal models . MAIN METHODS : A human CypA gene was fused with a protein transduction domain , PEP-1 peptide , to construct a cell permeable PEP-1-CypA protein . The protein expression level of cyclooxygenase-2 ( P35354 ) and cytokines was detected by Western blot , ELISA and mRNA level of P35354 and cytokines were measured by RT-PCR . The nuclear factor-kappa B ( NF-kB ) and mitogen-activated protein kinase ( MAPK ) activation were analyzed by Western blot and electrophoretic mobility shift assay . Skin inflammation was detected with immunohistochemistry . KEY FINDINGS : Transduced PEP-1-CypA protein markedly inhibited lipopolysaccharide- and 12-O-tetradecanoyl phorbol-13-acetate-induced expression levels of P35354 as well as pro-inflammatory cytokine levels in vitro and in vivo . Furthermore , transduced PEP-1-CypA protein resulted in a significant reduction in the activation of NF-kB and MAPK . SIGNIFICANCE : The results indicate that PEP-1-CypA inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK activation upon stimulation of inflammation in vitro and in vivo . PEP-1-CypA protein may potentially be used as a therapeutic agent against skin diseases-related inflammation . 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 .

MH_train_1496

MH_train_1496

MH_train_1496

interacts_with DB06779?

multiple_choice

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 . 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 . Thrombospondin 1 and its mimetic peptide DB05434 decrease angiogenesis and inflammation in a murine model of inflammatory bowel disease . OBJECTIVE : Vascular abnormalities and expression of proangiogenic factors have been repeatedly reported in inflammatory bowel disease ( Q9UKU7 ) . Thrombospondin 1 ( P07996 -1 ) is a protein well known for its antiangiogenic and anti-inflammatory properties . Using the dextran sulfate sodium ( DSS ) model , the role of P07996 -1 in Q9UKU7 has been investigated in vivo . METHODS : P07996 -1-deficient mice ( P07996 -1-/- ) and WT mice were treated with DSS for 7 days . Disease activity indices , myeloperoxidase activity ( P05164 ) and histology were analyzed . Microvascular density ( P53602 ) was quantified using immunohistochemistry ( IMH ) with CD31 antibody . TGF-beta(1) , basic FGF , P15692 , P01375 and MMPs protein levels were evaluated by IMH and enzyme-linked immunoabsorbent assay ( ELISA ) . Mice were treated with DB05434 ( Abbott Laboratories ) , an antiangiogenic P07996 peptide , using miniosmotic pumps for 7 days . RESULTS : P07996 -1(-/-) mice had a worse clinical outcome and exhibited severe signs of rectal bleeding compared to the WT controls . The P07996 -1-/- mice showed a higher level of crypt damage and deeper lesions . The grade of inflammation and the levels of P05164 activity were also significantly higher in colons of P07996 -1-/- mice . P07996 -1-/- mice displayed higher P53602 in focal areas of the colon after only 3 days of DSS treatment . Furthermore , clinical severity of the colitis and angiogenesis was significantly diminished when mice was treated with DB05434 . CONCLUSIONS : These findings directly link P07996 -1 as a protective factor in Q9UKU7 and suggest antiangiogenesis treatment , including compounds such as DB05434 as an adjuvant therapy for Q9UKU7 . 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 . 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 . 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 . Phase 1 trial of linifanib ( DB06080 ) in patients with refractory or relapsed acute myeloid leukemia . Linifanib , a potent oral inhibitor of fms-like tyrosine kinase 3 ( P36888 ) and vascular endothelial growth factor receptor tyrosine kinases , has demonstrated promising preclinical single-agent and synergistic anti-leukemic activity in combination with cytarabine . In this phase 1 , multicenter , open-label , dose-escalation study , 45 adults with relapsed/refractory acute myeloid leukemia ( AML ) received linifanib alone in arm A ( n = 29 ) and linifanib plus intermediate-dose cytarabine in arm B ( n = 16 ) . Median treatment duration was 21 days ( range 5-110 ) . Linifanib was well tolerated overall . The most common grade 3/4 events were fatigue ( arm A ) and febrile neutropenia ( arm B ) . The recommended phase 2 dose was 15 mg ( alone ) , and 10 mg ( with cytarabine ) . Evidence of on-target kinase inhibition in patients with P36888 -mutant and wild-type AML was seen . Decreased phosphorylated P36888 was seen in 3/3 patients with P36888 -internal tandem duplication ( ITD ) with peripheral blast reductions and in 8/24 ( 33 % ) patients with wild-type , D835 or unknown P36888 mutation . Eight/29 ( 28 % ) patients had decreased phosphorylated extracellular signal-regulated kinase ( P29323 ) . 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 . 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 . 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 . A cross-cultural study : anti-inflammatory activity of Australian and Chinese plants . In this study , in vitro inhibitory effects of 33 ethanol extracts obtained from 24 plant species ( representing 11 different families ) on cyclooxygenase-1 ( P23219 ) were evaluated . The plant materials selected for this study have been used in aboriginal medicine in Australia and traditional medicine in China for the treatment of various diseases that are considered as inflammation in nature , e.g. asthma , arthritis , rheumatism , fever , edema , infections , snakebite and related inflammatory diseases . All of the selected plants , with one exception , showed inhibitory activity against P23219 , which supports their traditional uses . The most potent P23219 inhibition were observed from the extracts of Acacia ancistrocarpa leaves ( IC(50)=23 microg/ml ) . Ficus racemosa bark , Clematis pickeringii stem , Acacia adsurgens leaves , Tinospora smilacina stem and Morinda citrifolia fruit powder exhibited inhibition of P23219 with the IC(50) of 100 , 141 , 144 , 158 and 163 microg/ml , respectively . DB00945 and indomethacin used as the reference P23219 inhibitors in this study inhibited P23219 with IC(50) of 241 and 1.2 microg/ml , respectively . The findings of this study may explain at least in part why these plants have been traditionally used for the treatment of inflammatory conditions in Australian aboriginal medicine and traditional Chinese medicine . 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 . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . 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 . Induction of RISK by P04035 inhibition affords cardioprotection after myocardial infarction . OBJECTIVES : Coronary occlusion and revascularization leads to myocardial damage and heart function deterioration . Statins can regress atherosclerosis and modulate platelet function , but their effect on post-acute myocardial infarction ( AMI ) injury remains to be fully determined . We sought to examine whether rosuvastatin ( R ) exerts any effect on the RISK/apoptosis pathway when administered early after coronary reperfusion . METHODS : Pigs were fed 10 days a hypercholesterolemic diet before AMI induction and thereafter for 7 days randomly distributed to receive R or placebo ( C ) with the same diet . At sacrifice , hearts were sliced and alternatively collected for MI size and molecular analysis ( gene and protein expression ) in the peri-infarcted and remote myocardium . The RISK components ( PKC , Erk2 , and Akt/ P31749 ) and downstream targets ( HIF-1alpha and P15692 ) , and cell survival/apoptosis markers ( Bcl-2 , Bax , and P42574 ) were analyzed . Annexin-V , Mito-Tracker staining , and inflammatory infiltration were also evaluated . RESULTS : R enhanced PKC , Erk2 , Akt/ P31749 and its downstream effectors , and attenuated inflammation and cardiomyocyte apoptosis in the peri-infarcted zone ( p < 0.05 ) . No changes were detected in the remote myocardium . Infarct size was smaller in R than in C pigs ( 7 % absolute reduction ; 36 % relative reduction ; p < 0.05 ) and was associated with an absolute 12 % recovery of LVEF ( 24 % relative restoration ; p < 0.05 vs. post-AMI ) . CONCLUSIONS : HMG- DB01992 inhibition early after reperfusion activates RISK kinases , reduces the extent of damaged myocardium , and improves heart function . P25116 expression is upregulated in prostate cancer . BACKGROUND : Aberrant expression of protease-activated receptors ( PARs ) has been associated with increased angiogenesis , tumor growth , and metastasis of various cancers . We assessed the status of PAR1 expression in prostate cancer . METHODS : The study compared the abundance levels of PAR1 RNA and protein using real-time reverse-transcriptase polymerase chain reaction and immunoblotting in freshly resected prostate tissues from early localized-stage disease ( n=9 ) to those from patients with advanced metastatic disease ( n=7 ) . PAR1 expression and localization was evaluated using immunohistochemical staining of prostate specimens with benign prostatic hyperplasia ( n=27 ) , early- ( n=32 ) and advanced-stage ( n=22 ) prostate cancer . Association analyses of PAR1 expression with expression of P15692 -family of growth factors , their receptors , and clinicopathological characteristics of the patients were also performed . RESULTS : PAR1 RNA expression in advanced-stage prostate was 2.39-fold higher ( P=0.024 ) and its protein expression was 2.75-fold higher ( P=5.89x10(-5) ) when compared with early-stage prostate cancer . PAR1 expression was localized to endothelial cells in vascular network of prostate tumor areas . The expression of PAR1 correlated statistically significantly with advanced disease stage ( P=0.0006 ) and pre-operative PSA levels ( P=0.005 ) in these samples . CONCLUSIONS : These findings demonstrate that PAR1 expression is increased in prostate cancer . Its predominant expression in vascular network suggests that it may play a direct and crucial role in angiogenesis and could be a relevant target for therapeutic interventions to control or to prevent disease progression .

MH_train_1497

MH_train_1497

MH_train_1497

interacts_with DB01576?

multiple_choice

The role of the polyol pathway in acute kidney injury caused by hindlimb ischaemia in mice . The polyol pathway , a collateral glycolytic process , previously considered to be active in high glucose milieu , has recently been proposed to play a crucial role in ischaemia/reperfusion tissue injury . In this study , we explored the role of the polyol pathway in acute kidney injury ( AKI ) , a life-threatening condition , caused by hindlimb ischaemia , and determined if inhibition of the polyol pathway by aldose reductase ( AR ) inhibitor is beneficial for this serious disorder . Mice 8 weeks of age rendered hindlimb ischaemic for 3 h by the clipping of major supporting arteries revealed marked muscle necrosis with accumulation of sorbitol and fructose in ischaemic muscles . Serum concentrations of blood urea nitrogen ( BUN ) , creatinine phosphokinase ( CPK ) , creatinine , tumour necrosis factor ( P01375 ) -alpha as well as interleukin ( IL ) -6 were all elevated in these mice . Treatment with AR inhibitor ( Q9Y4X5 ) effectively suppressed muscle necrosis and accompanying inflammatory reactions and prevented renal failure . Similar to Q9Y4X5 -treated mice , AR-deficient mice were protected from severe ischaemic limb injury and renal failure , showing only modest muscle necrosis and significant suppression of serum markers of renal failure and inflammation . Thus , these findings suggest that the polyol pathway is implicated in AKI caused by ischaemic limb injury and that AR may be a potential therapeutic target for this condition . DB00819 : future perspective in topical glaucoma therapeutics . Through this review it is contemplated that acetazolamide ( Q9Y6V0 ) , an age-old treatment for glaucoma with a myriad of side effects and inadequate topical effectiveness , may be formulated into a topically effective agent by utilizing various newer formulation approaches of ocular drug delivery . Even though it has a poor solubility and penetration power , various studies mentioned in the review indicate that it is possible to successfully formulate topically effective Q9Y6V0 by using : ( i ) high concentration of the drug , ( ii ) surfactant gel preparations of Q9Y6V0 , ( iii ) Q9Y6V0 loaded into liposomes , ( iv ) cyclodextrins to increase the solubility and hence bioavailability of Q9Y6V0 , and ( v ) viscolyzers and other polymers either alone or in combination with cyclodextrins . With the advent of newer topical carbonic anhydrase inhibitors ( CAIs ) like dorzolamide and brinzolamide , a localized effect with fewer side effects is expected . But whenever absorbed systemically , a similar range of adverse effects ( attributable to sulphonamides ) may occur upon use . Furthermore , oral Q9Y6V0 is reported to be more physiologically effective than 2 % dorzolamide hydrochloride administered topically , even though in isolated tissues dorzolamide appears to be the most active as it shows the lowest IC(50) values for P00918 and P22748 [ M.F. Surgue , J. Ocular Pharmacol. Ther. 12 ( 1996 ) 363-376 ] . Hence , there exists considerable scope for the development of more/equally effective and inexpensive topically effective formulations of Q9Y6V0 . The use of various formulation technologies discussed in this review can provide a fresh impetus to research in this area . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . 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 . Involvement of sphingosine kinase in P01375 -stimulated tetrahydrobiopterin biosynthesis in P13671 glioma cells . In P13671 glioma cells , the sphingolipid second messenger ceramide potentiates expression of inducible nitric-oxide synthase ( P35228 ) induced by tumor necrosis factor alpha ( P01375 ) without affecting P30793 ( GTPCH ) , the rate-limiting enzyme in the biosynthesis of 6(R)- DB00360 ( BH(4) ) , a cofactor required for P35228 activity . P01375 also stimulates sphingosine kinase , the enzyme that phosphorylates sphingosine to form sphingosine-1-phosphate ( Q8TCT9 ) , a further metabolite of ceramide . Several clones of P13671 cells , expressing widely varying levels of sphingosine kinase , were used to examine the role of Q8TCT9 in regulation of GTPCH and BH(4) biosynthesis . Overexpression of sphingosine kinase , with concomitant increased endogenous Q8TCT9 levels , potentiated the effect of P01375 on GTPCH expression and activity and BH(4) biosynthesis . In contrast , enforced expression of sphingosine kinase had no effect on P35228 expression or NO formation . Furthermore , N,N-dimethylsphingosine , a potent sphingosine kinase inhibitor , completely eliminated the increased GTPCH activity and expression induced by P01375 . Surprisingly , we found that , although P13671 cells can secrete Q8TCT9 , which is enhanced by P01375 , treatment of P13671 cells with exogenous Q8TCT9 or dihydro- Q8TCT9 had no affect on BH(4) biosynthesis . However , both Q8TCT9 and dihydro- Q8TCT9 markedly stimulated P29323 1/2 in P13671 cells , which express cell surface Q8TCT9 receptors . Interestingly , although this P29323 activation was blocked by PD98059 , which also reduced cellular proliferation induced by enforced expression of sphingosine kinase , PD98059 had no effect on GTPCH activity . Collectively , these results suggest that only intracellularly generated Q8TCT9 plays a role in regulation of GTPCH and BH(4) levels . Adaptive mutations in a human immunodeficiency virus type 1 envelope protein with a truncated V3 loop restore function by improving interactions with P01730 . We previously reported that a human immunodeficiency virus type 1 ( HIV-1 ) clade B envelope protein with a severely truncated V3 loop regained function after passage in tissue culture . The adapted virus , termed Q96RJ0 , retained the V3 truncation , was exquisitely sensitive to neutralization by the P01730 binding site monoclonal antibody b12 and by HIV-positive human sera , used P51681 to enter cells , and was completely resistant to small molecule P51681 antagonists . To examine the mechanistic basis for these properties , we singly and in combination introduced each of the 5 mutations from the adapted clone Q96RJ0 into the unadapted envelope . We found that single amino acid changes in the P01024 region , the V3 loop , and in the fusion peptide were responsible for imparting near-normal levels of envelope function to Q96RJ0 . T342A , which resulted in the loss of a highly conserved glycosylation site in P01024 , played the primary role . The adaptive amino acid changes had no impact on P51681 antagonist resistance but made virus more sensitive to neutralization by antibodies to the P01730 binding site , modestly enhanced affinity for P01730 , and made Q96RJ0 more responsive to P01730 binding . Specifically , Q96RJ0 was triggered by soluble P01730 more readily than the parental Env and , unlike the parental Env , could mediate entry on cells that express low levels of P01730 . In contrast , Q96RJ0 interacted with P51681 less efficiently and was highly sensitive to antibodies that bind to the P51681 N terminus and ECL2 . Therefore , enhanced utilization of P01730 is one mechanism by which HIV-1 can overcome mutations in the V3 region that negatively affect P51681 interactions . P15121 inhibition counteracts oxidative-nitrosative stress and poly(ADP-ribose) polymerase activation in tissue sites for diabetes complications . This study evaluated the effects of aldose reductase inhibition on diabetes-induced oxidative-nitrosative stress and poly(ADP-ribose) polymerase ( PARP ) activation . In animal experiments , control and streptozotocin-induced diabetic rats were treated with or without the aldose reductase inhibitor ( Q9Y4X5 ) fidarestat ( 16 mg . kg(-1) . day(-1) ) for 6 weeks starting from induction of diabetes . DB09391 pathway intermediate , but not glucose , accumulation in sciatic nerve and retina was completely prevented in diabetic rats treated with fidarestat . Sciatic motor nerve conduction velocity , hindlimb digital sensory nerve conduction velocity , and sciatic nerve concentrations of two major nonenzymatic antioxidants , glutathione and ascorbate , were reduced in diabetic versus control rats , and these changes were prevented in diabetic rats treated with fidarestat . DB02021 prevented the diabetes-induced increase in nitrotyrosine ( a marker of peroxynitrite-induced injury ) and poly(ADP-ribose) immunoreactivities in sciatic nerve and retina . DB02021 counteracted increased superoxide formation in aorta and epineurial vessels and in in vitro studies using hyperglycemia-exposed endothelial cells , and the DCF test/flow cytometry confirmed the endothelial origin of this phenomenon . DB02021 did not cause direct inhibition of PARP activity in a cell-free system containing PARP and NAD(+) but did counteract high-glucose-induced PARP activation in Schwann cells . In conclusion , aldose reductase inhibition counteracts diabetes-induced nitrosative stress and PARP activation in sciatic nerve and retina . These findings reveal the new beneficial properties of fidarestat , thus further justifying the ongoing clinical trials of this specific , potent , and low-toxic Q9Y4X5 . 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 . P50750 a potential target for drug development . The family of P12004 -Dependent Kinases ( CDKs ) can be subdivided into two major functional groups based on their roles in cell cycle and/or transcriptional control . P50750 is the catalytic subunit of positive transcription elongation factor b ( P-TEFb ) . P50750 is the kinase of the P50750 complex ( Tat-associated kinase complex ) , and binds to Tat protein of HIV , suggesting a possible role for P50750 in AIDS progression . P50750 complexed with its regulatory partner cyclin T1 , serves as a cellular mediator of the transactivation function of the HIV Tat protein . P-TEFb is responsible for the phosphorylation of the carboxyl-terminal domain of RNA Pol II , resulting in stimulation of transcription . Furthermore , the complexes containing P50750 induce the differentiation in distinct tissue . The P50750 /cyclin T1 complex is expressed at higher level in more differentiated primary neuroectodermal and neuroblastoma tumors , showing a correlation between the kinase expression and tumor differentiation grade . This may have clinical and therapeutical implications for these tumor types . Among the CDK inhibitors two have shown to be effective against P50750 : Roscovitine and DB03496 . These two inhibitors prevented the replication of human immunodeficiency virus ( HIV ) type 1 by blocking Tat transactivation of the HIV type 1 promoter . These compounds inhibit CDKs by binding to the catalytic domain in place of DB00171 , preventing transfer of a phosphate group to the substrate . More sensitive therapeutic agents of P50750 can be designed , and structural studies can add information in the understanding of this kinase . The major features related to P50750 inhibition will be reviewed in this article . Dopamine modulating drugs influence striatal (+)-[11C]DTBZ binding in rats : Q05940 binding is sensitive to changes in vesicular dopamine concentration . Binding of (+)-[11C]DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of (+)-[11C]DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , +14 % ) or DB01576 ( d- P49418 , 20 mg/kg , +12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , -16 % ) or levodopa ( -20 % ) . We suggest that in vivo (+)-[11C]DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed . 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 . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Strategies for the analysis of oncogene overexpression . Studies of the neu oncogene in breast carcinoma . The development of a consistent strategy for the analysis of oncogene expression at the cellular level is essential for understanding the roles of these genes in the development and progression of human neoplasia . Detection of the neu oncogene products in breast carcinoma was selected as a model for analysis of oncogene expression . Fifty-two primary human breast carcinomas were evaluated by quantitation of neu DNA amplification and mRNA expression and by localization of neu mRNA and protein ( p 185 ) at the cellular level by in situ hybridization ( ISH ) and immunohistochemistry ( IHC ) . The specificity and sensitivity of the molecular and immunologic probes for neu were established with the use of genetically engineered cell lines that overexpressed either neu or epidermal growth factor receptor ( P00533 ) . Twenty-nine percent of breast carcinomas demonstrated neu DNA amplification and mRNA overexpression , and there was close correlation between the level of neu mRNA expression and detection of neu gene products by ISH and IHC . Thirty-two percent of carcinomas demonstrated neu mRNA overexpression by ISH . The immunohistochemical method using Q96RJ0 monoclonal antibody for p185 was exquisitely sensitive in acetone-fixed frozen sections and provided an excellent approach for judging overexpression as confirmed by the various molecular analyses . All areas of nonmalignant breast epithelium stained weakly , and a wide range of staining intensity was observed in malignant breast epithelium , with 31 % of carcinomas judged to be p185 overexpressors . Heterogeneous expression of p185 was seen in some carcinomas . This study provides a strategic approach for the evaluation of oncogene expression in human tumors . Immunohistochemical study of the serotonergic neuronal system in an animal model of the mood disorder . The monoamine theory is one of the major hypotheses about the biological etiology of major depressive disorders . Recent pharmacological and postmortem investigations suggest that depressed patients have alterations in function of serotonergic neuronal system . However , the exact sites of alterations and the association between these alterations and the etiology of the disorder are still unclear . To elucidate these issues , we immunohistochemically examined vesicle monoamine transporter 2 ( Q05940 ) , serotonin receptor type 1a ( 5HT1a ) , and serotonin transporter ( P31645 ) in the hippocampal region of reserpine-treated rats , an animal model of depression . The results showed more Q05940 -immunoreactive varicose fibers in the pyramidal cell layer of hippocampus and parahippocampal cortexes , and more intense P31645 -immunoreactivity in the pyramidal cell layer and the area P22748 of hippocampus in the animal models compared to those of the controls . On the other hand , lower density of 5HT1a-immunoreactive deposits in the pyramidal cell layer of hippocampus and the parahippocampal cortex was observed in the animal models compared to those of the controls . These results suggest that a deficit of monoamines induces the alterations in the expression of the storage protein , the receptor and the transporter that are involved in the serotonergic neurotransmission in the hippocampal region . These alterations may underlie the changes of serotonergic system observed in the brains of patients with the depressive disorder . 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 . 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 . DB00428 diabetes and the expression of P11166 at the brush border and basolateral membranes of intestinal enterocytes . Changes in membrane expression of sodium-dependent glucose transporter ( P13866 ) and glucose transporter isoform ( P11168 ) protein have been implicated in the increased intestinal glucose transport in streptozotocin-diabetes . The possible involvement of P11166 in the transport response , however , has not previously been studied . Using confocal microscopy on tissue sections and Western blotting of purified brush border membrane ( BBM ) and basolateral membrane ( BLM ) , we have examined enterocyte expression of P11166 in untreated and in 1 and 21 day streptozotocin diabetic rats . In control enterocytes , P11166 was absent at the BBM and detected at low levels at the BLM . Diabetes resulted in a 4- to 5-fold increased expression of P11166 at the BLM and the protein could also be readily detected at the BBM . P01308 treatment of diabetic rats increased P11166 level at the BBM but was without effect on expression of the protein at the BLM . DB00050 , a gonadotropin-releasing hormone antagonist , induces the expression of melatonin receptor 1a in the gonadotropin-releasing hormone neuronal cell line GT1-7 . Melatonin has been implicated in the control of the reproductive system , and the modulatory actions of melatonin on gonadotropin-releasing hormone ( DB00644 ) neurons have been assumed to be indirectly mediated through afferent neurons . However , our previous studies demonstrate sexually dimorphic modulation of A-type gamma-aminobutyric acid ( GABA ) receptor ( GABA(A)R ) currents by melatonin in adult rat DB00644 neurons and a preferential expression of melatonin 1a receptor ( MT1 ) in male DB00644 neurons . Using immortalized DB00644 neurons ( GT1-7 cells ) , the present study investigated the mechanism by which the expression of melatonin receptors is regulated in DB00644 neurons . Like endogenous DB00644 neurons , GT1-7 cells express both DB00644 and P30968 mRNAs , indicating that the cells have a self-stimulatory system . A 2-iodomelatonin binding assay and RT-PCR analysis demonstrated that the cells expressed neither MT1 nor P02795 . However , treatment of GT1-7 cells with the DB00644 antagonist cetrorelix significantly increased 2-iodomelatonin binding and induced a time- and concentration-dependent MT1 mRNA expression . The GABA(A)R currents were then measured using a perforated patch-clamp technique to examine whether the treatment with cetrorelix changed the responses to melatonin . Melatonin augmented the GABA(A)R currents in GT1-7 cells treated with 1 muM cetrorelix for 24 h , while melatonin decreased the currents in the cells not treated with cetrorelix , probably via receptor-independent processes . The present results suggest that DB00644 downregulates the expression of MT1 via an autocrine-paracrine mechanism in GT1-7 cells , and modifies the melatonin-induced modulation of GABA(A)R currents . These findings may provide one possible mechanism for the sexually dimorphic responses to melatonin in adult rat DB00644 neurons . Cdk5/p25(nck5a) interaction with synaptic proteins in bovine brain . P12004 -dependent kinase 5 ( Cdk5 ) exists in large multimeric complexes , but its function and binding partners in these complexes are unclear . We explored these issues by chromatographic and immunochemical analyses of Cdk5 and p25(nck5a) ( a neuronal Cdk5 activator ) and their associated proteins from bovine brain . Mono-S column enzyme eluates were divided into three fractions and analyzed by gel filtration . The majority of p25(nck5a) from Mono-S fractions I , II , and III eluted from the gel filtration column at approximately 60 , 200 , and 400 kDa , respectively , and Cdk5 was abundant in fractions > 400 kDa . We characterized these macromolecular structures by immunoprecipitating p25(nck5a) , followed by a second immunoprecipitation of remaining unbound proteins using a Cdk5 antibody . The p25(nck5a) immunoprecipitates showed association with Cdk5 . P49418 was detected in the 400-kDa complex and synapsin I in the > 400 kDa structure . The Cdk5 immunoprecipitates , however , revealed abundant retained Cdk5 but no remaining p25(nck5a) , indicating that Cdk5 in macromolecular structures is mostly unassociated with p25(nck5a) . Thus , we demonstrate : an amphiphysin-associated 400-kDa Cdk5/p25(nck5a) complex , a synapsin I-associated > 400-kDa Cdk5/p25(nck5a) complex , and nck5a-free Cdk5 complexes ( 200 to > 400 kDa ) . P49418 acts as a Cdk5/p25(nck5a) substrate in the 400-kDa complex and we speculate that Cdk5/p25(nck5a) participates in amphiphysin-mediated endocytosis . Activation of nuclear factor-kappaB-dependent transcription by tumor necrosis factor-alpha is mediated through phosphorylation of RelA/p65 on serine 529 . Nuclear factor-kappaB ( NF-kappaB ) is an essential transcription factor in the control of expression of genes involved in immune and inflammatory responses . In unstimulated cells , NF-kappaB complexes are sequestered in the cytoplasm through interactions with P25963 and other IkappaB proteins . Extracellular stimuli that activate NF-kappaB , such as tumor necrosis factor alpha ( TNFalpha ) , cause rapid phosphorylation of P25963 at serines 32 and 36 . The inducible phosphorylation of P25963 is followed by its ubiquitination and degradation , allowing NF-kappaB complexes to translocate into the nucleus and to activate gene expression . Previously , it has been shown that TNFalpha as well as other stimuli also lead to the phosphorylation of the RelA/p65 subunit of NF-kappaB . In this report , we demonstrate that the TNFalpha-induced phosphorylation of the RelA/p65 subunit occurs on serine 529 , which is in the C-terminal ( Q96RJ0 ) transactivation domain . Accordingly , the TNFalpha-induced phosphorylation of Rel/p65 increases NF-kappaB transcriptional activity but does not affect nuclear translocation or DNA binding affinity . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . A novel approach in the treatment of cancer : targeting the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) autocrine pathway contributes to a number of processes important to cancer development and progression , including cell proliferation , apoptosis , angiogenesis , and metastatic spread . The critical role the P00533 plays in cancer has led to an extensive search for selective inhibitors of the P00533 signaling pathway . The results of a large body of preclinical studies and the early clinical trials thus far conducted suggest that targeting the P00533 could represent a significant contribution to cancer therapy . A variety of different approaches are currently being used to target the P00533 . The most promising strategies in clinical development include monoclonal antibodies to prevent ligand binding and small molecule inhibitors of the tyrosine kinase enzymatic activity to inhibit autophosphorylation and downstream intracellular signaling . At least five blocking monoclonal antibodies have been developed against the P00533 . Among these , IMC-225 is a chimeric human-mouse monoclonal IgG1 antibody that has been the first anti- P00533 targeted therapy to enter clinical evaluation in cancer patients in Phase II and III studies , alone or in combination with conventional therapies , such as radiotherapy and chemotherapy . A number of small molecule inhibitors of the P00533 tyrosine kinase enzymatic activity is also in development . DB00530 and ZD1839 ( DB00317 ) are currently in Phase II and III development , respectively . ZD1839 , a p.o. active , selective quinazoline derivative has demonstrated promising in vitro and in vivo antitumor activity . Preliminary results from Phase I and II trials in patients with advanced disease demonstrate that ZD1839 and DB00530 have an acceptable tolerability profile and promising clinical efficacy in patients with a variety of tumor types . This mini-review describes the P00533 inhibitors in clinical development .

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interacts_with DB09053?

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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 . 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 . Collision-induced dissociation of valdecoxib metabolites : a novel rearrangement involving an isoxazole ring . DB00580 is a potent P35354 inhibitor . During metabolism studies of valdecoxib by liquid chromatography/tandem mass spectrometry , we observed a novel mass spectral rearrangement involving an isoxazole ring for some of the metabolites in the negative ion mode . Accurate mass measurements were performed with quadrupole time-of-flight mass spectrometry to determine the elemental compositions of the fragments . Additionally , two types of stable-isotope labeled analogues were prepared to assist with the assignments of these fragments and possible mechanistic rearrangements resulting from collision-induced dissociation ( CID ) . Detailed analyses of the CID mass spectra suggest that the fragmentation process involves a novel two-step rearrangement . The first step consists of an intramolecular Q8WUX1 reaction with a five-membered ring rearrangement to form an intermediate . The second step involves a four-membered ring intramolecular rearrangement followed by a cleavage of the N-O bond on the isoxazole ring to form a unique fragment ion at m/z 196 . The same phenomenon was observed for a group of structurally related metabolites that also contain a 5-hydroxymethyl or 5-carboxylic acid moieties . A mechanism for the novel rearrangement involving an isoxazole ring is proposed . Integration of porcine chromosome 13 maps . In order to expand the comparative map between human chromosome 3 ( Q9NZP2 ) and porcine chromosome 13 ( SSC13 ) , seven genes from Q9NZP2 were mapped on SSC13 by fluorescence in situ hybridisation ( Q5TCZ1 ) , viz . P09110 , P15309 , O60513 , P02788 , Q15746 , P11177 and P10826 . With a view to integrating this expanded comparative map with the existing SSC13 linkage map , we used the INRA-University of Minnesota porcine Radiation Hybrid panel ( IMpRH ) to localize more precisely and to order 15 genes on the SSC13 map , viz . P15309 , O95622 , P05090 , P06276 , P42081 , P35462 , P17677 , P05166 , P04049 , P08100 , SI , TF , P02786 , Q02880 and Q9UQR1 . In this way , we were able to create an integrated map , containing 38 type I and 81 type II markers , by correlating the linkage , radiation hybrid ( RH ) and cytogenetic maps of SSC13 . This integrated map will give us the opportunity to take maximal advantage of the comparative mapping strategy for positional candidate cloning of genes responsible for economically important traits . DB11320 Promotes the Release of P05231 via the P35367 /p38 and NF-κB Pathways in Nasal Fibroblasts . PURPOSE : Based on the close relationship between histamine and interleukin 6 ( P05231 ) , we hypothesized that histamine may regulate the production of cytokines , such as P05231 , during allergic inflammation . Here , we examined the role of histamine in P05231 production and histamine receptor activity in nasal fibroblasts , along with the mechanisms underlying these effects . METHODS : Experiments were performed using nasal fibroblasts from 8 normal patients . RT-PCR was used to identify the major histamine receptors expressed in nasal fibroblasts . Fibroblasts were then treated with histamine with or without histamine-receptor antagonists , and monitored for P05231 production using an ELISA . Four potential downstream signaling molecules , p38 , extracellular signal-regulated kinase ( P29323 ) , c-Jun N-terminal kinase ( JNK ) , and NF-κB , were evaluated by Western blot , and a luciferase reporter assay . RESULTS : Elevated expression was seen for all histamine receptors , with P05231 protein levels increasing significantly following histamine stimulation . Among the histamine-receptor specific antagonists , only the P35367 antagonist significantly decreased P05231 production in histamine-stimulated nasal fibroblasts . DB11320 increased the expression level of phosphorylated p38 ( pp38 ) , pERK , and pJNK , as well as NF-κB induction . The P35367 antagonist actively suppressed pp38 and NF-κB expression in histamine-induced nasal fibroblasts , but not pERK and pJNK . The p38 inhibitor strongly attenuated P05231 production in histamine-stimulated nasal fibroblasts . CONCLUSIONS : The data presented here suggest that antihistamines may be involved in the regulation of cytokines , such as P05231 , due to the role of histamine as an inflammatory mediator in nasal fibroblasts . Human bronchial smooth muscle cells express adenylyl cyclase isoforms 2 , 4 , and 6 in distinct membrane microdomains . Adenylyl cyclases ( AC ) are important regulators of airway smooth muscle function , because β-adrenergic receptor ( AR ) agonists stimulate AC activity and increase airway diameter . We assessed expression of AC isoforms in human bronchial smooth muscle cells ( hBSMC ) . Reverse transcriptase-polymerase chain reaction and immunoblot analyses detected expression of AC2 , AC4 , and AC6 . DB02587 -stimulated AC activity in membranes from hBSMC displayed Ca(2+)-inhibited and G(βγ)-stimulated AC activity , consistent with expression of AC6 , AC2 , and AC4 . DB01064 -stimulated AC activity was inhibited by Ca(2+) but unaltered by G(βγ) , whereas butaprost-stimulated AC activity was stimulated by G(βγ) but unaffected by Ca(2+) addition . Using sucrose density centrifugation to isolate lipid raft fractions , we found that only AC6 localized in lipid raft fractions , whereas AC2 and AC4 localized in nonraft fractions . Immunoisolation of caveolae using caveolin-1 antibodies yielded Ca(2+)-inhibited AC activity ( consistent with AC6 expression ) , whereas the nonprecipitated material displayed G(βγ)-stimulated AC activity ( consistent with expression of AC2 and/or AC4 ) . Overexpression of AC6 enhanced DB02527 production in response to isoproterenol and beraprost but did not increase responses to prostaglandin E(2) or butaprost . β(2)AR , but not prostanoid EP(2) or EP(4) receptors , colocalized with O95622 /6 in lipid raft fractions . Thus , particular G protein-coupled receptors couple to discreet AC isoforms based , in part , on their colocalization in membrane microdomains . These different DB02527 signaling compartments in airway smooth muscle cells are responsive to different hormones and neurotransmitters and can be regulated by different coincident signals such as Ca(2+) and G(βγ) . Phosphodiesterase 4B mediates extracellular signal-regulated kinase-dependent up-regulation of mucin P98088 protein by Streptococcus pneumoniae by inhibiting DB02527 -protein kinase A-dependent P28562 phosphatase pathway . Otitis media ( OM ) is the most common childhood bacterial infection and the major cause of conductive hearing loss in children . Mucus overproduction is a hallmark of OM . Streptococcus pneumoniae is the most common gram-positive bacterial pathogen causing OM . Among many mucin genes , P98088 has been found to be greatly up-regulated in the middle ear mucosa of human patients with OM . We previously reported that S. pneumoniae up-regulates P98088 expression in a MAPK P29323 -dependent manner . We also found that MAPK phosphatase-1 ( P28562 ) negatively regulates S. pneumoniae-induced P29323 -dependent P98088 up-regulation . Therapeutic strategies for up-regulating the expression of negative regulators such as P28562 may have significant therapeutic potential for treating mucus overproduction in OM . However , the underlying molecular mechanism by which P28562 expression is negatively regulated during S. pneumoniae infection is unknown . In this study we show that phosphodiesterase 4B ( Q07343 ) mediates S. pneumoniae-induced P98088 up-regulation by inhibiting the expression of a negative regulator P28562 , which in turn leads to enhanced MAPK P29323 activation and subsequent up-regulation of P98088 . Q07343 inhibits P28562 expression in a DB02527 -PKA-dependent manner . DB05876 -specific inhibitor rolipram inhibits S. pneumoniae-induced P98088 up-regulation both in vitro and in vivo . Moreover , we show that Q07343 plays a critical role in P98088 induction . Finally , topical and post-infection administration of rolipram into the middle ear potently inhibited S. pneumoniae-induced P98088 up-regulation . Collectively , these data demonstrate that Q07343 mediates P29323 -dependent up-regulation of mucin P98088 by S. pneumoniae by inhibiting DB02527 -PKA-dependent P28562 pathway . This study may lead to novel therapeutic strategy for inhibiting mucus overproduction . [ 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 . 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 . Inhibition of cyclooxygenases 1 and 2 by the phospholipase-blocker , arachidonyl trifluoromethyl ketone . BACKGROUND AND PURPOSE : Arachidonyl trifluoromethyl ketone ( Q06187 ) is widely used as an inhibitor of cytosolic group IV phospholipase A(2) ( cPLA(2) ) and calcium-independent group VI phospholipase A(2) ( iPLA(2) ) . Q06187 thus reduces arachidonic acid ( AA ) substrate for cyclooxygenase ( P36551 ; also known as prostaglandin H synthase ) and attenuates prostaglandin ( PG ) synthesis . It has been shown previously , that Q06187 blocks thromboxane B(2) production induced by exogenous AA in human platelets . It remains , however , unknown whether Q06187 also directly modulates the activity of cyclooxygenase ( P36551 ) . EXPERIMENTAL APPROACH : Time courses for inhibition of P36551 by Q06187 was obtained using osteoblast-like MC3T3-E1 cells , with exogenous AA as substrate and the pure enzymes P23219 and P35354 . PGE(2) was measured by GC-MS . KEY RESULTS : Q06187 was a potent inhibitor of P23219 and P35354 with IC(50) values of 0.5 and 0.1 microM in MC3T3-E1 cells and of 1.7 and 2.6 microM using the pure enzymes . Inhibition was reversible , with slow- and tight-binding characteristics . The arachidonyl carbon chain was essential , as the saturated palmitoyl analogue had no effect . CONCLUSIONS AND IMPLICATIONS : Attenuation of PG synthesis by Q06187 is taken to be the consequence of PLA(2) inhibition and the findings of many studies are interpreted on that basis . If there are , however , alternative routes for AA liberation ( such as phospholipase C/diacyl glycerol lipase or phospholipase D ) , this interpretation can lead to false conclusions . As Q06187 is a widely used and important pharmacological tool in eicosanoid research , knowledge of its interactions with other major enzymes of the cascade is of considerable importance . A mechanism for the synergistic antimalarial action of atovaquone and proguanil . A combination of atovaquone and proguanil has been found to be quite effective in treating malaria , with little evidence of the emergence of resistance when atovaquone was used as a single agent . We have examined possible mechanisms for the synergy between these two drugs . While proguanil by itself had no effect on electron transport or mitochondrial membrane potential ( DeltaPsim ) , it significantly enhanced the ability of atovaquone to collapse DeltaPsim when used in combination . This enhancement was observed at pharmacologically achievable doses . DB01131 acted as a biguanide rather than as its metabolite cycloguanil ( a parasite dihydrofolate reductase [ P00374 ] inhibitor ) to enhance the atovaquone effect ; another P00374 inhibitor , pyrimethamine , also had no enhancing effect . DB01131 -mediated enhancement was specific for atovaquone , since the effects of other mitochondrial electron transport inhibitors , such as myxothiazole and antimycin , were not altered by inclusion of proguanil . Surprisingly , proguanil did not enhance the ability of atovaquone to inhibit mitochondrial electron transport in malaria parasites . These results suggest that proguanil in its prodrug form acts in synergy with atovaquone by lowering the effective concentration at which atovaquone collapses DeltaPsim in malaria parasites . This could explain the paradoxical success of the atovaquone-proguanil combination even in regions where proguanil alone is ineffective due to resistance . The results also suggest that the atovaquone-proguanil combination may act as a site-specific uncoupler of parasite mitochondria in a selective manner . DB09053 treatment ameliorates murine chronic graft-versus-host disease . Chronic graft-versus-host disease ( cGVHD ) is a life-threatening impediment to allogeneic hematopoietic stem cell transplantation , and current therapies do not completely prevent and/or treat cGVHD . P01730 + T cells and B cells mediate cGVHD ; therefore , targeting these populations may inhibit cGVHD pathogenesis . DB09053 is an FDA-approved irreversible inhibitor of Bruton 's tyrosine kinase ( Q06187 ) and P60568 inducible T cell kinase ( Q08881 ) that targets Th2 cells and B cells and produces durable remissions in B cell malignancies with minimal toxicity . Here , we evaluated whether ibrutinib could reverse established cGVHD in 2 complementary murine models , a model interrogating T cell-driven sclerodermatous cGVHD and an alloantibody-driven multiorgan system cGVHD model that induces bronchiolar obliterans ( BO ) . In the T cell-mediated sclerodermatous cGVHD model , ibrutinib treatment delayed progression , improved survival , and ameliorated clinical and pathological manifestations . In the alloantibody-driven cGVHD model , ibrutinib treatment restored pulmonary function and reduced germinal center reactions and tissue immunoglobulin deposition . Animals lacking Q06187 and Q08881 did not develop cGVHD , indicating that these molecules are critical to cGVHD development . Furthermore , ibrutinib treatment reduced activation of T and B cells from patients with active cGVHD . Our data demonstrate that B cells and T cells drive cGVHD and suggest that ibrutinib has potential as a therapeutic agent , warranting consideration for cGVHD clinical trials . MiR-24 tumor suppressor activity is regulated independent of p53 and through a target site polymorphism . MicroRNAs ( miRNAs ) are predicted to regulate approximately 30 % of all human genes ; however , only a few miRNAs have been assigned their targets and specific functions . Here we demonstrate that miR-24 , a ubiquitously expressed miRNA , has an anti-proliferative effect independent of p53 function . Cell lines with differential p53 status were used as a model to study the effects of miR-24 on cell proliferation , cell cycle control , gene regulation and cellular transformation . Overexpression of miR-24 in six different cell lines , independent of p53 function , inhibited cell proliferation and resulted in G2/S cell cycle arrest . MiR-24 over expression in cells with wt-p53 upregulated P04637 and P38936 protein ; however , in p53-null cells miR-24 still induced cell cycle arrest without the involvement of P38936 . We show that miR-24 regulates p53-independent cellular proliferation by regulating an S-phase enzyme , dihydrofolate reductase ( P00374 ) a target of the chemotherapeutic drug methotrexate ( MTX ) . Of interest , we found that a miR-24 target site polymorphism in P00374 3' UTR that results in loss of miR-24-function and high P00374 levels in the cell imparts a growth advantage to immortalized cells and induces neoplastic transformation . Of clinical significance , we found that miR-24 is deregulated in human colorectal cancer tumors and a subset of tumors has reduced levels of miR-24 . A novel function for miR-24 as a p53-independent cell cycle inhibitory miRNA is proposed . Raloxifene concurrently stimulates osteoprotegerin and inhibits interleukin-6 production by human trabecular osteoblasts . Raloxifene reduces bone loss and prevents vertebral fractures in postmenopausal women . Its skeletal effects are mediated by estrogen receptors ( ER ) and their modulation of paracrine osteoblastic factors . Receptor activator of nuclear factor-kappa B ligand is essential for osteoclasts and enhances bone resorption , whereas osteoprotegerin ( O00300 ) neutralizes receptor activator of nuclear factor-kappa B ligand . Here , we assessed the effects of raloxifene on O00300 production in human osteoblasts ( hOB ) . Raloxifene enhanced gene expression of P03372 and progesterone receptor . Moreover , raloxifene increased O00300 mRNA levels and protein secretion by hOB in a dose- and time-dependent fashion by 2- to 4-fold with a maximum effect at 10(-7) M and after 72 h ( P < 0.001 ) . Treatment with the ER antagonist DB00947 abrogated the effects of raloxifene on O00300 production . Moreover , raloxifene enhanced osteoblastic differentiation markers , type 1 collagen secretion , and alkaline phosphatase activity by 3- and 2-fold , respectively ( P < 0.001 ) . In addition , raloxifene inhibited expression of the bone-resorbing cytokine P05231 by 25-45 % ( P < 0.001 ) . In conclusion , our data suggest that raloxifene stimulates O00300 production and inhibits P05231 production by hOB . Because O00300 production increases with osteoblastic maturation , enhancement of O00300 production by raloxifene could be related to its stimulatory effects on osteoblastic differentiation . 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 . DB06273 infusion therapy normalizes inflammation in sporadic P35858 patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 ) through caspase1 , interleukin 1 ( IL1 ) , P05231 and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 receptor ( P08887 ) . DB06273 inhibits global interleukin-6 ( P05231 ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra(R) ) infusions . At baseline , one half of P35858 subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4-fold , P < 0.05 vs. controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 , P80098 , Q99616 and O00175 . DB06273 infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 concentrations of tocilizumab inhibited caspase1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up- or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions . 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 . 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 . PDE4B5 , a novel , super-short , brain-specific DB02527 phosphodiesterase-4 variant whose isoform-specifying N-terminal region is identical to that of DB02527 phosphodiesterase-4D6 ( PDE4D6 ) . The DB02527 -specific phosphodiesterase-4 ( DB05876 ) gene family is the target of several potential selective therapeutic inhibitors . The four DB05876 genes generate several distinct protein-coding isoforms through the use of alternative promoters and 5'-coding exons . Using mouse transcripts , we identified a novel , super-short isoform of human Q07343 encoding a novel 5' terminus , which we label PDE4B5 . The protein-coding region of the novel 5' exon is conserved across vertebrates , chicken , zebrafish , and fugu . Reverse-transcription-polymerase chain reaction ( PCR ) and quantitative ( PCR ) measurements show that this isoform is brain-specific . The novel protein is 58 +/- 2 kDa ; it has DB02527 hydrolyzing enzymatic activity and is inhibited by DB05876 -selective inhibitors rolipram and cilomilast ( DB03849 ) . Confocal and subcellular fractionation analyses show that it is distributed predominantly and unevenly within the cytosol . The 16 novel N-terminal residues of PDE4B5 are identical to the 16 N-terminal residues of the super-short isoform of Q08499 ( PDE4D6 ) , which is also brain-specific . PDE4B5 is able to bind the scaffold protein Q9NRI5 , whose gene has been linked to schizophrenia . Microarray expression profiling of the DB05876 gene family shows that specific DB05876 genes are enriched in muscle and blood fractions ; however , only by monitoring the individual isoforms is the brain specificity of the super-short Q08499 and Q07343 isoforms revealed . Understanding the distinct tissue specificity of DB05876 isoforms will be important for understanding phosphodiesterase biology and opportunities for therapeutic intervention . 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 . Toll-like receptor signaling is impaired in dendritic cells from patients with X-linked agammaglobulinemia . Bruton 's tyrosine kinase ( Q06187 ) , which is defective in patients with X-linked agammaglobulinemia ( XLA ) , is expressed not only in B cells but also in monocytes and dendritic cells ( DCs ) . DCs play a crucial role in the innate immune response against infections by sensing pathogens through Toll-like receptors ( TLRs ) . However , it is not known whether Q06187 deficiency in XLA might impair TLR-mediated signaling in DCs , which are susceptible to various infections . The phenotypic maturation and cytokine production mediated by TLRs were examined in monocyte-derived DC from XLA patients and normal controls . The TLR expression in DCs was analyzed by flow cytometry . TLR-mediated signaling in DCs was evaluated for the phenotypic maturation based on Q01151 expression and production of cytokines , such as P01375 , P05231 and IL-12p70 . TLR levels in DCs were similar between XLA and controls . O60603 , O00206 and Q9NYK1 /8 ligands elicited less phenotypic maturation of DCs from XLA patients than normal controls based on Q01151 expression . Stimulation with O60603 , O00206 and Q9NYK1 /8 ligands , as well as O15455 ligand , resulted in significantly lower production of P01375 , but neither P05231 nor IL-12p70 , by DCs from XLA patients in comparison to normal controls . These findings suggest that Q06187 may thus be required for TLR signaling in DCs . The impaired TLR signaling in DCs may therefore be partly responsible for the occurrence of severe infections with bacteria and some viruses in XLA patients .

MH_train_1499

MH_train_1499

MH_train_1499

interacts_with DB01171?

multiple_choice

DB01257 reduces complement activation , inflammation , endothelial damage , thrombosis , and renal injury markers in aHUS . Atypical hemolytic uremic syndrome ( aHUS ) is a genetic , life-threatening disease characterized by uncontrolled complement activation , systemic thrombotic microangiopathy ( TMA ) , and vital organ damage . We evaluated the effect of terminal complement blockade with the anti- P01031 monoclonal antibody eculizumab on biomarkers of cellular processes involved in TMA in patients with aHUS longitudinally , during up to 1 year of treatment , compared with in healthy volunteers . Biomarker levels were elevated at baseline in most patients , regardless of mutational status , plasma exchange/infusion use , platelet count , or lactate dehydrogenase or haptoglobin levels . DB01257 reduced terminal complement activation ( C5a and sC5b-9 ) and renal injury markers ( clusterin , cystatin-C , β2-microglobulin , and liver fatty acid binding protein-1 ) to healthy volunteer levels and reduced inflammation ( soluble tumor necrosis factor receptor-1 ) , coagulation ( prothrombin fragment F1+2 and d-dimer ) , and endothelial damage ( thrombomodulin ) markers to near-normal levels . Alternative pathway activation ( Ba ) and endothelial activation markers ( soluble vascular cell adhesion molecule-1 ) decreased but remained elevated , reflecting ongoing complement activation in aHUS despite complete terminal complement blockade . These results highlight links between terminal complement activation and inflammation , endothelial damage , thrombosis , and renal injury and underscore ongoing risk for systemic TMA and progression to organ damage . Further research regarding underlying complement dysregulation is warranted . This trial was registered at www.clinicaltrials.gov as # NCT01194973 . 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 . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . A 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 . Effects of interleukin-2 ( P60568 ) on human plasma lipid , lipoprotein , and P02741 . Six patients with confirmed malignant disease received four consecutive weekly cycles of human recombinant interleukin-2 ( P60568 ) 4 days/week , continuous iv. infusion , 3 X 10(6) U/m2/day . Plasma cholesterol decreased a mean of 7 % within 24 hours after P60568 infusion and decreased by 33 % within 4 days . Plasma cholesterol was significantly lower than baseline concentration by day 21 ( -21 % ) , and day 25 ( -41 % ) was significantly lower than day 21 . Decreased plasma cholesterol was the result of decreased HDL and LDL cholesterol concentrations . Plasma triglyceride demonstrated a mean increase of 46 % after 4 days of therapy and remained greater than baseline concentrations at all time points analyzed . Apolipoprotein AI and AII decreased concomitantly with HDL-cholesterol concentrations , whereas apolipoprotein B after an initial mean decrease of 17 % during the first cycle was not significantly different from baseline during the fourth cycle . P02649 and P08519 were not significantly affected by P60568 treatment . Plasma P02741 ( CRP ) increased by 79 % within 24 hours of therapy , increased by 254 % on day 4 , then decreased to baseline concentrations by day 21 after 3 days off of P60568 . Day 25 CRP was elevated compared to both baseline and day 21 concentrations . P60568 induced plasma lipoprotein changes may be due in part to the induction of interferon gamma . Delirium and the functional recovery of older medical inpatients after acute illness : the significance of biological factors . Previous studies have not clarified the relationship of delirium to functional capacity during acute illness . We have investigated this relationship , incorporating the potential roles of P02649 genotype and circulating cytokines in a longitudinal study of acutely admitted patients aged 70+ years . In all participants was measured the : Barthel Index ( BI ) , mini-mental state examination ( MMSE ) , confusion assessment method ( P62158 ) , delirium rating scale ( Q9H307 ) , APACHE II , P02649 genotype . In a sub-sample : serum interferon-γ ( IFN-γ ) , interleukin-1 ( Levels of IL-1α , IL-1β and IL-1 receptor antagonist activity IL-1RA ) , interleukin-6 ( P05231 ) , leukemia inhibitory factor ( P15018 ) , tumor necrosis factor-α ( P01375 -α ) and insulin-like growth factor-I ( P05019 ) . Of 164 participants , mean age 84.6 ± 6.57 years ( ± S.D. ) , 67.1 % were women . On first assessment , mean BI was 14.13 ± 4.46 and delirium prevalence was 25.6 % . At discharge , the mean BI of survivors ( n=150 ) was 15.61 ± 4.22 . By discharge , survivors who had recovered from prevalent delirium had significant improvement in BI ( n=38 , p=0.005 ) , but non-recovers did not ( n=14 , p=0.512 ) . On , multivariate analysis , BI was significantly affected by MMSE , P02649 , IL-1α , P05231 , P15018 and P01375 -α levels ( p < 0.05 ) but not by delirium . Delirium in acutely admitted patients is associated with functional decline only in those who do not recover . Biological factors , rather that delirium itself , may be responsible for this . 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 . Interaction of early environment , gender and genes of monoamine neurotransmission in the aetiology of depression in a large population-based Finnish birth cohort . Objectives Depression is a worldwide leading cause of morbidity and disability . Genetic studies have recently begun to elucidate its molecular aetiology . The authors investigated candidate genes of monoamine neurotransmission and early environmental risk factors for depressiveness in the genetically isolated population-based Northern Finland Birth Cohort 1966 ( 12 058 live births ) . Design The authors ascertained and subdivided the study sample ( n=5225 ) based on measures of early development and of social environment , and examined candidate genes of monoamine neurotransmission , many of which have shown prior evidence of a gene-environment interaction for affective disorders , namely P31645 , Q8IWU9 , P21964 , P21397 and the dopamine receptor genes P21728 - P21918 . Results and conclusion The authors observed no major genetic effects of the analysed variants on depressiveness . However , when measures of early development and of social environment were considered , some evidence of interaction was observed . Allelic variants of P21964 interacted with high early developmental risk ( p=0.005 for rs2239393 and p=0.02 for rs4680 ) so that the association with depression was detected only in individuals at high developmental risk group ( p=0.0046 and β=0.056 for rs5993883-rs2239393-rs4680 risk haplotype CGG including Val158 ) , particularly in males ( p=0.0053 and β=0.083 for the haplotype CGG ) . Rs4274224 from P14416 interacted with gender ( p=0.017 ) showing a significant association with depressiveness in males ( p=0.0006 and β=0.0023 ; p=0.00005 and β=0.069 for rs4648318-rs4274224 haplotype GG ) . The results support the role of genes of monoamine neurotransmission in the aetiology of depression conditional on environmental risk and sex , but not direct major effects of monoaminergic genes in this unselected population . 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 . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . 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 . Multiple pathways of apolipoprotein E signaling in primary neurons . P02649 is a genetic risk factor for Alzheimer 's disease , and the apoE protein is associated with beta-amyloid deposits in Alzheimer 's disease brain . We examined signaling pathways stimulated by apoE in primary neurons in culture . ApoE and an apoE-derived peptide activated several intracellular kinases , including prominently extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . P27361 /2 activation by apoE was blocked by an inhibitor of the low-density lipoprotein receptor family , the specific DB01221 glutamate receptor antagonist MK 801 and other calcium channel blockers . Activation of apoE receptors also induced tyrosine phosphorylation of Dab1 , an adaptor protein of apoE receptors , but experiments in Dab1 knockout neurons demonstrated that Dab1 was not necessary for P29323 activation . In contrast , apoE treatment of primary neurons decreased activation of c-Jun N-terminal kinase , a kinase that interacts with another apoE receptor adaptor protein , c-Jun N-terminal kinase-interacting protein . This change also depended on interactions with the low-density lipoprotein receptor family but was independent of calcium channels . c-Jun N-terminal kinase deactivation by apoE was blocked by gamma-secretase inhibitors and pertussis toxin . These results demonstrate that apoE affects several signaling cascades in neurons : increased disabled phosphorylation , activation of the P27361 /2 pathway ( dependent on calcium influx via the DB01221 receptor ) and inhibition of the P45983 /2 pathway ( dependent on gamma-secretase and G proteins ) . Lack of allelic association of dopamine D1 and D2 ( TaqIA ) receptor gene polymorphisms with reduced dopaminergic sensitivity to alcoholism . Our study tested the hypothesis of whether the sensitivity of central dopamine receptors corresponds to the genotypic constitution of DNA-polymorphisms of the dopamine D1 and D2 receptor ( P21728 , P14416 ) genes and is associated with poor treatment outcome . Therefore , 97 alcohol-dependent patients were assessed according to their sensitivity of central dopamine receptors ( apomorphine-induced secretion of growth hormone ) , clinical outcome during a 6-month observation period , and genotypic constitution of the TaqIA restriction fragment length polymorphism ( RFLP ) at the P14416 locus and of the Bsp1286I RFLP at the P21728 locus . On the 1st day of detoxification , dopamine receptor hyposensitivity was found in treatment nonresponders , but not in responders . DB00714 -induced growth hormone release did not differ significantly in alcoholics with different genotypes of the P21728 and P14416 RFLPs . Neither did we find a significant allelic association with treatment response . Thus , we did not find evidence for a genetic determination of dopamine receptor hyposensitivity in alcoholics with poor treatment outcome . Selenate enhances P40763 transcriptional activity in endothelial cells : differential actions of selenate and selenite on P15018 cytokine signaling and cell viability . Sodium selenate may have utility in treating Alzheimer 's disease and diabetes ; however , its impact on the associated proinflammatory cytokine signaling of endothelial cells has not been investigated . We report that treatment of human microvascular endothelial cells with sodium selenate at a pharmacological dose ( 100 μM ) enhanced tyrosine phosphorylation of nuclear P40763 on Y705 in response to P05231 -type cytokine , leukemia inhibitory factor ( P15018 ) , indicative of enhanced P40763 activity . Accordingly , P40763 nuclear binding to DNA was increased , as well as P15018 -induced gene expression of chemokine ( C-C motif ) ligand 2 ( P13500 ) . P13500 plays a key role in inflammatory processes associated with neuronal degenerative and vascular diseases . The enhancing action of selenate on P15018 -induced P40763 Y705 phosphorylation was replicated by vanadate and a specific inhibitor of protein tyrosine phosphatase , non-receptor type 1 ( P18031 ) . Moreover , we observed that selenite , the cellular reduction bioproduct of selenate but not selenate itself , inhibited enzymatic activity of human recombinant P18031 . Our findings support the conclusion that in human microvascular endothelial cells selenate has a vanadate-like effect in inhibiting P18031 and enhancing proinflammatory P40763 activation . These findings raise the possibility that beneficial actions of supranutritional levels of selenate for treating Alzheimer 's and diabetes may be offset by a proinflammatory action on endothelial cells . Enhanced abdominal aortic aneurysm formation in thrombin-activatable procarboxypeptidase B-deficient mice . OBJECTIVE : To determine whether procarboxypeptidase B ( Q96IY4 ) (-/-) mice are susceptible to accelerated abdominal aortic aneurysm ( AAA ) development secondary to unregulated P10451 -mediated mural inflammation in the absence of P15086 inhibition . METHODS AND RESULTS : Thrombin/thrombomodulin cleaves thrombin-activatable Q96IY4 or thrombin-activatable fibrinolysis inhibitor , activating P15086 , which inhibits the generation of plasmin and inactivates proinflammatory mediators ( complement C5a and thrombin-cleaved osteopontin [ P10451 ] ) . P02649 (-/-) P10451 (-/-) mice are protected from experimental AAA formation . Murine AAAs were created via intra-aortic porcine pancreatic elastase ( PPE ) infusion . Increased mortality secondary to AAA rupture was observed in Q96IY4 (-/-) mice at the standard PPE dose . At reduced doses of PPE , Q96IY4 (-/-) mice developed larger AAAs than wild-type controls ( 1.01+/-0.27 versus 0.68+/-0.05 mm ; P=0.02 [ mean+/-SD ] ) . P01031 (-/-) and P10451 (-/-) mice were not protected against AAA development . Treatment with tranexamic acid inhibited plasmin generation and abrogated enhanced AAA progression in Q96IY4 (-/-) mice . CONCLUSIONS : This study establishes the role of P15086 in experimental AAA disease , indicating that P15086 has a broad anti-inflammatory role in vivo . Enhanced AAA formation in the PPE model is the result of increased plasmin generation , not unregulated C5a- or P10451 -mediated mural inflammation . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . 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 . Aging and age-related diseases -- from endocrine therapy to target therapy . Aging represents an important health issue not only for the individual , but also for society in general . Burdens associated with aging are expanding as longevity increases . This has led to an enhanced focus on issues related to aging and age-related diseases . Until recently , anti-aging endocrine-therapy has been largely limited to hormone-replacement therapy ( HRT ) that is associated with multiple side effects , including an increased risk of cancer . This has greatly limited the application of HRT in anti-aging therapy . Recently , the focus of anti-aging research has expanded from endocrine signaling pathways to effects on regulatory gene networks . In this regard , the P01286 -GH- DB01277 / P01308 , TOR- P23443 ,NAD(+)-Sirtuin , P04637 , Q9UEF7 and P02649 pathways have been linked to processes associated with age-related diseases , including cancer , cardiovascular disease , diabetes , osteoporosis , and neurodegenerative diseases , all of which directly influence health in aging , and represent key targets in anti-aging therapy . 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 . 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 .