interacts_with DB00173?
Anti-thrombin therapy during warm ischemia and cold preservation prevents chronic kidney graft fibrosis in a P81605 model . Ischemia reperfusion injury ( IRI ) is pivotal for renal fibrosis development via peritubular capillaries injury . Coagulation represents a key mechanism involved in this process . Melagatran ( M ) , a thrombin inhibitor , was evaluated in an autotransplanted kidney model , using Large White pigs . To mimic deceased after cardiac death donor conditions , kidneys underwent warm ischemia ( WI ) for 60 min before cold preservation for 24 h in University of Wisconsin solution . Treatment with M before WI and/or in the preservation solution drastically improved survival at 3 months , reduced renal dysfunction related to a critical reduction in interstitial fibrosis , measured by Sirius Red staining . Tissue analysis revealed reduced expression of transforming growth factor-beta ( TGF-beta ) and activation level of its effectors phospho- P84022 , Q13485 and connective tissue growth factor ( P29279 ) after M treatment . Fibrinolysis activation was also observed , evidenced by downregulation of P05121 protein and gene expression . In addition , M reduced P26447 expression and vimentin staining , which are markers for epithelial mesenchymal transition , a major pathway to chronic kidney fibrosis . Finally , expression of oxidative stress markers Nox2 and P35228 was reduced . We conclude that inhibition of thrombin is an effective therapy against IRI that reduces chronic graft fibrosis , with a significantly positive effect on survival . The murine chemokine receptor P61073 is tightly regulated during T cell development and activation . We have characterized the murine homolog of the HIV-co-receptor P61073 during T cell development and activation . Our data demonstrate that this chemokine receptor , although highly conserved between human and mouse , is differently expressed and regulated in both species . Mitogenic activation resulted in an increase of surface P61073 on murine T cells within 2 days , whereas the receptor was strongly down-regulated on human T cells during this period . Furthermore , intraperitoneal immunization of mice resulted in a strong increase of splenic and mesenteric cytotoxic T cells co-expressing P61073 . It is interesting that , on thymocytes , expression of P61073 is restricted to P01730 +CD8+ cells . Stromal cell-derived factor-1alpha , a natural ligand of P61073 , induced chemotaxis of thymocytes and was found to counteract dexamethasone-induced apoptosis to a certain extent in these cells . Thus , our data show that expression of P61073 is tightly controlled on murine T cells and indicate that this highly conserved chemokine receptor might serve different functions in humans and mice . Lack of significant effects of the type 2 diabetes susceptibility loci Q86VZ6 , O75794 / Q8IU85 , Q04721 , Q9P2N4 , Q6YHU6 , and P19075 / O75473 on diabetes and quantitative metabolic traits . Recently , several novel loci reaching genome-wide significance levels for type 2 diabetes ( T2D ) were identified through a meta-analysis of three genome-wide scans and large-scale follow-up . The aim of our study was to investigate the association of these loci with T2D and related subphenotypes in two cohorts from Germany . We performed an association study of 9 SNPs in or around Q86VZ6 , O75794 / Q8IU85 , Q04721 , Q9H165 , Q9P2N4 , P15692 , P81605 , Q6YHU6 , and P19075 / O75473 with T2D and related quantitative traits ( fasting insulin and glucose , indices derived from OGTT ) in the isolated population of Sorbs ( 205 cases and 695 controls ) and in a mixed German population ( Leipzig ) ( 938 subjects with and 918 without T2D ) . None of the variants was associated with T2D , but the meta-analysis of both cohorts revealed a modest trend of association of rs7578597 in Q6YHU6 with T2D ( p=0.055 ) . Furthermore , Sorbian subjects homozygous for the rs7578597 T-allele had lower mean 30-minute plasma insulin when compared with carriers of the C-allele ( p < 0.05 ) . The T-allele was also nominally associated with higher fasting plasma glucose in the Leipzig cohort ( p < 0.05 ) . Although several other SNPs showed some evidence for association with T2D-related traits the effects were not replicated within our study . Associations of the T2D-risk alleles with T2D or related subphenotypes were overall very weak in the approximately 2 700 subjects studied . This is compatible with the modest effect size of these " second sweep " variants , which will require large-scale association studies on quantitative traits to clarify their role in the pathophysiology of T2D . 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 . 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 . 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 . 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 . Forced dimerization increases the activity of Δ P00533 /EGFRvIII and enhances its oncogenicity . Delta epidermal growth factor receptor ( Δ P00533 ) , an in-frame deletion mutant of the extracellular ligand-binding domain , which occurs in about 30 % of glioblastoma , is a potent oncogene that promotes tumor growth and progression . The signaling of Δ P00533 is ligand-independent and low intensity , allowing it to evade the normal mechanisms of internalization and degradation by the endocytic machinery and hence is persistent . The basis of the oncogenic potential of Δ P00533 remains incompletely understood , including whether dimerization plays an important role in its signal and whether its oncogenic potential is dependent on its relatively low intensity , when compared with the acutely activated wild-type receptor . To examine these two important questions , we have generated a chimeric Δ P00533 that allows forced dimerization via domains derived from variants of the P62942 protein that are brought together by FK506 derivatives . Forced dimerization of chimeric Δ P00533 significantly increased the intensity of its signal , as measured by receptor phosphorylation levels , suggesting that the naturally occurring Δ P00533 does not form strong or stable dimers as part of its low level signal . Interestingly , the increased activity of dimerized , chimeric Δ P00533 did not promote receptor internalization , implying that reduced rate of endocytic downregulation of Δ P00533 is an inherent characteristic . Significantly , forced dimerization enhanced the oncogenic signal of the receptor , implying that the Δ P00533 is a potent oncogene despite , not because of its low intensity . 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 . 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 . Hyperbaric oxygen therapy reduces neuroinflammation and expression of matrix metalloproteinase-9 in the rat model of traumatic brain injury . The acute inflammatory response plays an important role in secondary brain damage after traumatic brain injury ( TBI ) . Neutrophils provide the main source of matrix metalloproteinases ( MMPs ) which also play a deleterious role in TBI . Numerous preclinical studies have suggested that hyperbaric oxygen therapy ( HBOT ) may by beneficial in various noncerebral and cerebral inflammatory diseases . The goal of this study was to evaluate the effects of HBOT on inflammatory infiltration and the expression of MMPs in correlation with secondary cell death in the rat model of dynamic cortical deformation ( P81605 ) . Twenty animals underwent P81605 with subsequent HBOT ( 2.8 ATA , two sessions of 45 min each ) ; 10 animals : P81605 and normobaric oxygenation ( 1 ATA ) , 10 animals : not treated after P81605 . Cell death was evaluated by TUNEL . Neutrophils were revealed by myeloperoxidase staining . Immunohistochemical staining for P08253 and -9 and tissue inhibitors of P03956 ( P01033 ) and -2 was also performed and the results were quantitatively evaluated by image analysis . In the animals treated by HBOT , a significant decrease in the number of TUNEL-positive cells and neutrophilic inflammatory infiltration was seen in comparison with nontreated animals and those treated by normobaric oxygen . The expression of P14780 was also significantly lower in the treated group . Staining for P08253 and P16035 did not change significantly . Our results demonstrate that HBOT decreased the extent of secondary cell death and reactive neuroinflammation in the TBI model . The decline of P14780 expression after HBOT may also contribute to protection of brain tissue in the perilesional area . Further research should be centred on the evaluation of long-term functional and morphological results of HBOT . Gene promoter methylation in colorectal cancer and healthy adjacent mucosa specimens : correlation with physiological and pathological characteristics , and with biomarkers of one-carbon metabolism . We evaluated the promoter methylation levels of the P25054 , P16455 , hMLH1 , RASSF1A and CDKN2A genes in 107 colorectal cancer ( CRC ) samples and 80 healthy adjacent tissues . We searched for correlation with both physical and pathological features , polymorphisms of folate metabolism pathway genes ( P42898 , Q9UBK8 , Q99707 , RFC1 , P04818 , and Q9UBC3 ) , and data on circulating folate , vitamin B12 and homocysteine , which were available in a subgroup of the CRC patients . An increased number of methylated samples were found in CRC respect to adjacent healthy tissues , with the exception of P25054 , which was also frequently methylated in healthy colonic mucosa . Statistically significant associations were found between RASSF1A promoter methylation and tumor stage , and between hMLH1 promoter methylation and tumor location . Increasing age positively correlated with both hMLH1 and P16455 methylation levels in CRC tissues , and with P25054 methylation levels in the adjacent healthy mucosa . Concerning gender , females showed higher hMLH1 promoter methylation levels with respect to males . In CRC samples , the Q99707 2756AG genotype correlated with higher methylation levels of RASSF1A , and the P04818 1494 6bp ins/del polymorphism correlated with the methylation levels of both P25054 and hMLH1 . In adjacent healthy tissues , Q99707 2756AG and P04818 1494 6bp del/del genotypes correlated with P25054 and P16455 promoter methylation , respectively . Low folate levels were associated with hMLH1 hypermethylation . Present results support the hypothesis that DNA methylation in CRC depends from both physiological and environmental factors , with one-carbon metabolism largely involved in this process . 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 . Nox-2 is a modulator of fibrogenesis in kidney allografts . We studied the role of classical phagocytic NADPH oxidase ( Nox ) in the pathogenesis of kidney allograft tubulointerstitial fibrosis . Immunofluorescence studies showed that Nox-2 and P13498 ( electron transfer subunits of Nox ) colocalized in the tubulointerstitium of human kidney allografts . Tubular Nox-2 also colocalized with alpha-SMA in areas of injury , suggestive of epithelial-to-mesenchymal transition ( EMT ) . Interstitial macrophages ( P34810 (+) ) and myofibroblasts ( alpha-SMA(+) ) expressed Nox-2 while graft infiltrating T cells ( CD3(+) ) and mature fibroblasts ( P26447 (+) ) were Nox-2(-) . These results were confirmed in the Fisher-to-Lewis rat kidney transplant model . Areas of tubulitis were associated with Nox-2 and alpha-SMA , suggestive of EMT . Immunoblot analyses showed that Nox-2 upregulation was associated with oxidative stress ( nitrotyrosine ) and fibrogenesis ( alpha-SMA and phospho- Q15796 ) at 3 weeks and 6 months . Allografts treated with Nox inhibitors ( DPI or apocynin ) for 1 week showed reduced fibronectin and phospho- Q15796 and increased P12830 levels . DB00091 A , TGF-beta1 and angiotensin II increased Nox-2 mRNA levels 2- to 7-fold in vitro ( NRK52E cells ) . Treatment with specific Nox inhibitors ( DPI or apocynin ) prevented the downregulation of P12830 and upregulation of fibronectin transcripts . In aggregate , these studies suggest that Nox-2 is involved in the pathogenesis of allograft tubulointerstitial fibrosis via activation transcription factor Q15796 , EMT and myofibroblasts . DB00134 recycling pathways and antimalarial drug design . 5'-Deoxy-5'-(methylthio)adenosine ( MTA ) is an S-adenosylmethionine metabolite that is generated as a by-product of polyamine biosynthesis . In mammalian cells , MTA undergoes a phosphorolytic cleavage catalyzed by Q13126 to produce adenine and 5-deoxy-5-(methylthio)ribose-1-phosphate ( Q15012 ) . DB00173 is utilized in purine salvage pathways , and Q15012 is subsequently recycled to methionine . Whereas some microorganisms metabolize MTA to Q15012 via Q13126 , others metabolize MTA to Q15012 in two steps via initial cleavage by MTA nucleosidase to adenine and 5-deoxy-5-(methylthio)ribose ( Q99707 ) followed by conversion of Q99707 to Q15012 by Q99707 kinase . In order to assess the extent to which these pathways may be operative in Plasmodium falciparum , we have examined a series of 5'-alkyl-substituted analogs of MTA and the related Q99707 analogs and compared their abilities to inhibit in vitro growth of this malarial parasite . The Q99707 analogs 5-deoxy-5-(ethylthio)ribose and 5-deoxy-5-(hydroxyethylthio)ribose were inactive at concentrations up to 1 mM , and 5-deoxy-5-(monofluoroethylthio)ribose was weakly active ( 50 % inhibitory concentration = 700 microM ) . In comparison , the MTA analogs , 5'-deoxy-5'-(ethylthio)adenosine,5'-deoxy-5'-(hydroxyethylthio)ade nosine ( HETA ) , and 5'-deoxy-5'-(monofluoroethylthio)adenosine , had 50 % inhibitory concentrations of 80 , 46 , and 61 microM , respectively . Extracts of P. falciparum were found to have substantial Q13126 activity . Coadministration of MTA with HETA partially protected the parasites against the growth-inhibitory effects of HETA . Results of this study indicate that P. falciparum has an active Q13126 that can be targeted by analogs of MTA . Endothelial cell transforming growth factor-β receptor activation causes tacrolimus-induced renal arteriolar hyalinosis . Arteriolar hyalinosis is a common histological finding in renal transplant recipients treated with the calcineurin inhibitor tacrolimus ; however , the pathophysiologic mechanisms remain unknown . In addition to increasing transforming growth factor ( TGF ) -β levels , tacrolimus inhibits calcineurin by binding to FK506-binding protein 12 ( P62942 ) . P62942 alone also inhibits TGF-β receptor activation . Here we tested whether tacrolimus binding to P62942 removes an inhibition of the TGF-β receptor , allowing ligand binding , ultimately leading to receptor activation and arteriolar hyalinosis . We found that specific deletion of P62942 from endothelial cells was sufficient to activate endothelial TGF-β receptors and induce renal arteriolar hyalinosis in these knockout mice , similar to that induced by tacrolimus . DB00864 -treated and knockout mice exhibited significantly increased levels of aortic TGF-β receptor activation as evidenced by Q15796 /3 phosphorylation , along with increased collagen and fibronectin expression compared to controls . Treatment of isolated mouse aortas with tacrolimus increased TGF-β receptor activation and collagen and fibronectin expression . These effects were independent of calcineurin , absent in endothelial denuded aortic rings , and could be prevented by the small molecule TGF-β receptor inhibitor SB-505124 . Thus , endothelial cell TGF-β receptor activation is sufficient to cause vascular remodeling and renal arteriolar hyalinosis . Comparison of diafiltration and tangential flow filtration for purification of nanoparticle suspensions . PURPOSE : The study reports evaluation of different purification processes for removing surplus surfactant and formulating stable nanoparticle dispersions . METHODS : Nanoparticle formulations prepared from poly(D,L-lactide-co-glycolide) and polyvinyl alcohol ( P32926 ) were purified by a diafiltration centrifugal device ( P81605 ) , using 300K and 100K molecular weight cut-off ( MWCO ) membranes and a tangential flow filtration ( TFF ) system with a 300K MWCO membrane . The effects of process parameters including MWCO , transmembrane pressure ( P54849 ) , and mode of TFF on nanoparticle purification were evaluated , and two purification techniques were compared to the commonly used ultracentrifugation technique . RESULTS : Both P81605 and TFF systems ( concentration mode at P54849 of 10 psi ) with 300K MWCO membrane removed maximal percent P32926 from nanoparticle dispersions ( 89.0 and 90.7 % , respectively ) . T90 , the time taken to remove 90 % of P32926 in 200-ml sample , however , was considerably different ( 9.6 and 2.8 h , respectively ) . Purified nanoparticle dispersions were stable and free of aggregation at ambient conditions over 3 days . This is in contrast to the ultracentrifugation technique , which , although it can yield a highly purified sample , suffers from drawbacks of a level of irreversible nanoparticle aggregation and loss of fine particles in the supernatant during centrifugation . CONCLUSIONS : The TFF , in concentration mode at P54849 of 10 psi , is a relatively quick , efficient , and cost-effective technique for purification and concentration of a large nanoparticle batch ( > or=200 ml ) . The P81605 technique can be an alternative purification method for nanoparticle dispersions of small volumes . Platelet aggregation inhibition in patients receiving statins either fully or partially metabolized by P08684 . DB00758 therapy is the standard for prevention of cardiovascular thrombotic events . DB00758 is converted to an active thiol by the cytochrome P450 CYP 3A4 and 2C19 enzymes . Recent studies suggest that statins metabolized by P08684 attenuate the anti-aggregatory effect of clopidogrel . We evaluated the effect of P08684 -metabolized statins ( atorvastatin , group 1 ) and partially- P08684 -metabolized statins ( simvastatin , group 2 ) on platelet aggregation inhibition ( P05121 ) when given concomitantly with clopidogrel as compared to patients who were statin naive ( group 3 ) . P05121 was measured by PlateletWorks ( Helena Laboratories ICHOR ) using the platelet Q9H244 receptor agonist ADP ( 20 micromol ) . All patients were on clopidogrel therapy ( 75 mg/day ) . Non-responsiveness was defined as a P05121 of < 35 % . There was no statistical difference in mean P05121 among groups ; a higher prevalence of clopidogrel non-responders was noted in group 1 compared to group 3 ( p=0.002 ) . Multivariate analysis , adjusting for unequal presence of metabolic syndrome and hypertension , we found no statistical difference between groups . Our data suggests that statins , either fully or partially metabolized by P08684 , do not influence P05121 when clopidogrel is used at 75 mg/day , even after adjusting for risk factors . We concluded that concomitant statins with clopidogrel therapy does not influence the effect of clopidogrel in P05121 . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Angiotensin converting enzyme-2 confers endothelial protection and attenuates atherosclerosis . The endothelium plays a central role in the maintenance of vascular homeostasis . One of the main effectors of endothelial dysfunction is P03950 II , and pharmacological approaches to limit P03950 II bioactivity remain the cornerstone of cardiovascular therapeutics . Angiotensin converting enzyme-2 ( Q9BYF1 ) has been identified as a critical negative modulator of P03950 II bioactivity , counterbalancing the effects of P12821 in determining net tissue P03950 II levels ; however , the role of Q9BYF1 in the vasculature remains unknown . In the present study , we hypothesized that Q9BYF1 is a novel target to limit endothelial dysfunction and atherosclerosis . To this aim , we performed in vitro gain and loss of function experiments in endothelial cells and evaluated in vivo angiogenesis and atherosclerosis in apolipoprotein E-knockout mice treated with AdACE2 . Q9BYF1 -deficient mice exhibited impaired endothelium-dependent relaxation . Overexpression of Q9BYF1 in human endothelial cells stimulated endothelial cell migration and tube formation , and limited monocyte and cellular adhesion molecule expression ; effects that were reversed in Q9BYF1 gene silenced and endothelial cells isolated from Q9BYF1 -deficient animals . Q9BYF1 attenuated P03950 II-induced reactive oxygen species production in part through decreasing the expression of P13498 . The effects of Q9BYF1 on endothelial activation were attenuated by pharmacological blockade of P03950 -(1-7) with A779 . Q9BYF1 promoted capillary formation and neovessel maturation in vivo and reduced atherosclerosis in apolipoprotein E-knockout mice These data indicate that Q9BYF1 , in an P03950 -(1-7)-dependent fashion , functions to improve endothelial homeostasis via a mechanism that may involve attenuation of NADPHox-induced reactive oxygen species production . Q9BYF1 -based treatment approaches may be a novel approach to limit aberrant vascular responses and atherothrombosis . Redox signaling in angiogenesis : role of NADPH oxidase . Angiogenesis , a process of new blood vessel formation , is a key process involved in normal development and wound repair as well as in the various pathophysiologies such as ischemic heart and limb diseases and atherosclerosis . Reactive oxygen species ( ROS ) such as superoxide and H(2)O(2) function as signaling molecules in many aspects of growth factor-mediated responses including angiogenesis . Vascular endothelial growth factor ( P15692 ) is a key angiogenic growth factor and stimulates proliferation , migration , and tube formation of endothelial cells ( ECs ) primarily through the P15692 receptor type2 ( VEGR2 , P35968 /Flk1 ) . P15692 binding initiates autophosphorylation of P35968 , which results in activation of downstream signaling enzymes including P27361 /2 , Akt , and P29474 in ECs , thereby stimulating angiogenesis . The major source of ROS in EC is a NADPH oxidase which consists of Nox1 , Nox2 ( gp91phox ) , Nox4 , P13498 , p47phox , p67phox and the small G protein Rac1 . The endothelial NADPH oxidase is activated by angiogenic factors including P15692 and angiopoietin-1 . ROS derived from this enzyme stimulate diverse redox signaling pathways leading to angiogenesis-related gene induction as well as EC migration and proliferation , which may contribute to postnatal angiogenesis in vivo . The aim of this review is to provide an overview of the recent progress on the emerging area of the role of ROS derived from NADPH oxidase and redox signaling in angiogenesis . Understanding these mechanisms may provide insight into the NADPH oxidase and redox signaling components as potential therapeutic targets for treatment of angiogenesis-dependent cardiovascular diseases and for promoting angiogenesis in ischemic limb and heart diseases . Genetic aspects of ischemic stroke : coagulation , homocysteine , and lipoprotein metabolism as potential risk factors . Stroke is one of the most common causes of death and long term disability throughout the world . It may be the outcome of a number of monogenic disorders or , more commonly , a polygenic multifactorial disease . Numerous studies have investigated the role of genetics in the pathogenesis of ischemic stroke , with varied and often contradictory results . The candidate ' stroke risk ' genes affecting haemostasis ( P12259 , F2 , P02671 / P02675 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / P08514 , P17301 , P07359 , TPA , Q96IY4 , P07204 , PZ , P08758 ) , homocysteine metabolism ( P42898 , P35520 , Q99707 ) , and lipid metabolism ( apo E , P06858 , P11597 , O95477 , apo AI , apo CIII , apo AIV , apo AV , apo B , apo H , apo(a) , P27169 /2/3 , P01130 / P78380 ) are evaluated in this review . By examining meta-analyses and case-control studies , we made a classification of gene/gene polymorphisms according to the degree of association with ischemic stroke risk . The data assembled could be very useful for further meta-analysis and for future clinical applications . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . 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 novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Enhanced sensitivity to irinotecan by Cdk1 inhibition in the p53-deficient HT29 human colon cancer cell line . Mutations in the tumor-suppressor gene p53 have been associated with advanced colorectal cancer ( CRC ) . DB00762 ( CPT-11 ) , a P11387 inhibitor , has been recently incorporated to the adjuvant therapy . Since the DNA-damage checkpoint depends on p53 activation , the status of p53 might critically influence the response to CPT-11 . We analysed the sensitivity to CPT-11 in the human colon cancer cell line HT29 ( mut p53 ) and its wild-type (wt)-p53 stably transfected subclone HT29-A4 . Cell-cycle analysis in synchronised cells demonstrated the activation of transfected wt-p53 and a P38936 ( P38936 /CIP1)-dependent cell-cycle blockage in the S phase . Activated wt-p53 increased apoptosis and enhanced sensitivity to CPT-11 . In p53-deficient cells , cDNA-macroarray analysis and western blotting showed an accumulation of the cyclin-dependent kinase (cdk)1/cyclin B complex . Subsequent p53-independent activation of the cdk-inhibitor ( cdk-I ) P38936 ( P38936 /CIP1) prevented cell-cycle progression . Cdk1 induction was exploited in vivo to improve the sensitivity to CPT-11 by additional treatment with the cdk-I P99999 -202 . We demonstrate a gain of sensitivity to CPT-11 in a p53-mutated colon cancer model either by restoring wild-type p53 function or by sequential treatment with cdk-Is . Considering that mutations in p53 are among the most common genetic alterations in CRC , a therapeutic approach specifically targeting p53-deficient tumors could greatly improve the treatment outcomes . DB00087 induction and triple maintenance immunotherapy in kidney transplantation from donors after cardiac death . We have used alemtuzumab in combination with triple maintenance immunosuppression in renal transplantation from donors after cardiac death between 2002 and 2006 . We compared outcomes of induction therapy with alemtuzumab with interleukin-2 ( P60568 ) receptor antagonists ( RA ) and anti-lymphocyte antibodies . We used a retrospective sequential study design to examine 170 recipients of kidneys from donor after cardiac death ( P81605 ) for survival , graft survival , time to first rejection , glomerular filtration and complications . Patients were stratified into high-risk and low-risk groups based on the following criteria : panel of reactive antibodies > 20 % , retransplants , Afro-American race . Induction with alemtuzumab was compared with anti-thymocyte globulin ( ATG ) in the high-risk and with IL-2RA in the low-risk group . Patients received triple immunosuppression with steroids , mycophenolate mofetil and calcineurin inhibitors . Patient survival , graft survival , rejection rate and glomerular filtration rate did not significantly differ between patients treated with alemtuzumab versus IL-2RAs or ATG . There was a trend towards reduced graft- and patient survival in the alemtuzumab group . There was an increased incidence of cytomegalovirus ( CMV ) infections in the alemtuzumab-induced group and a trend towards increased BK virus and bacterial infections . Induction of P81605 kidney transplants with alemtuzumab compared to IL-2RA and ATG has no significant impact on acute rejection . It appears however that CMV infections are increased in patients induced with alemtuzumab . We therefore conclude that induction with alemtuzumab does not confer any advantage over traditional induction agents . 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 . Examination of the role of magnetic resonance imaging in multiple sclerosis : A problem-orientated approach . Magnetic Resonance Imaging ( Q9BWK5 ) has brought in several benefits to the study of Multiple Sclerosis ( MS ) . It provides accurate measurement of disease activity , facilitates precise diagnosis , and aid in the assessment of newer therapies . The imaging guidelines for MS are broadly divided in to approaches for imaging patients with suspected MS or clinically isolated syndromes ( Q9NSE2 ) or for monitoring patients with established MS . In this review , the technical aspects of MR imaging for MS are briefly discussed . The imaging process need to capture the twin aspects of acute MS viz. the autoimmune acute inflammatory process and the neurodegenerative process . Gadolinium enhanced Q9BWK5 can identify acute inflammatory lesions precisely . The commonly applied Q9BWK5 marker of disease progression is brain atrophy . Whole brain magnetization Transfer Ratio ( Q99707 ) and Magnetic Resonance Spectroscopy ( P59665 ) are two other techniques use to monitor disease progression . A variety of imaging techniques such as Double Inversion Recovery ( P30518 ) , Spoiled Gradient Recalled ( SPGR ) acquisition , and Fluid Attenuated Inversion Recovery ( FLAIR ) have been utilized to study the cortical changes in MS . Q9BWK5 is now extensively used in the Phase I , II and III clinical trials of new therapies . As the technical aspects of Q9BWK5 advance rapidly , and higher field strengths become available , it is hoped that the impact of Q9BWK5 on our understanding of MS will be even more profound in the next decade . Synergistic inhibition of breast cancer cell lines with a dual inhibitor of P00533 -HER-2/neu and a Bcl-2 inhibitor . The epidermal growth factor receptor ( P00533 ) ( ErbB1 ) and HER-2/neu ( ErbB2 ) are members of the ErbB family of receptor tyrosine kinases . These receptors are overexpressed in a variety of human tumors and overexpression generally correlates with poor prognosis and decreased survival . DB01259 , a reversible inhibitor of both P00533 and HER-2/neu , has shown some success in achieving clinical responses in heavily pretreated advanced cancer patients . GW2974 is a reversible dual inhibitor similar to lapatinib , but GW2974 was not progressed to clinical trials due to pharmacokinetic issues . Bcl-2 , an anti-apoptotic protein , is also overexpressed in a number of human tumors . Bcl-2 inhibitors induce apoptosis and sensitize cancer cells to other therapies . The purpose of this study was to assess the effects of combining ErbB and Bcl-2 inhibitors on the growth of human breast cancer cell lines . P00533 /HER-2/neu tyrosine kinase inhibitors ( lapatinib and GW2974 ) were combined with Bcl-2 inhibitors ( HA14-1 or GX15-070 ) and the anti-proliferative effects were determined by the MTT tetrazolium dye assay . Combinations were tested in MCF-7 human breast cancer cells , a HER-2/neu transfected MCF-7 cell line ( MCF/18 ) , and a tamoxifen-resistant MCF-7 cell line ( Q99707 -3 ) . A synergistic inhibitory effect was observed with the combination of inhibitors of P00533 -HER-2/neu ( lapatinib or GW2974 ) and Bcl-2 ( GX15-070 or HA14-1 ) on the growth of the MCF-7 , MCF/18 , and Q99707 -3 human breast cancer cell lines . This study suggests that simultaneously blocking the ErbB family of receptor tyrosine kinases and Bcl-2 family of proteins may be a benefit to breast cancer patients . Molecular markers of programmed cell death in donor hearts before transplantation . BACKGROUND : In this study we investigate whether pro-apoptotic , pro-inflammatory and other early signaling markers indicative of increased propensity for cell death processes were evident in human donor heart allografts immediately before transplantation , and whether there is an association with primary graft failure . METHODS : A prospective study was performed utilizing donor left atrial myocardium collected at the time of implantation of hearts from brain-dead donors ( BDD , n = 29 ) . In addition , to explore the potential of donor hearts from donation after circulatory death ( P81605 ) , myocardial samples were obtained during transplantation of lungs from P81605 donors ( n = 6 ) . A comparator reference group ( n = 7 ) consisted of left atrial specimens from patients undergoing mitral valve surgery . RESULTS : Significantly raised levels of caspase-3 specific activity , activated hypoxia inducible factor-1 ( HIF-1α ) and 8-hydroxy-2'-deoxyguanosine were evident in the transplanted hearts ( from BDD ) that developed primary graft failure ( n = 11 ) . P81605 hearts did not differ from BDD with regard to mRNA expression levels of FAS , Bax , P05231 and caspase-3 . Although P81605 hearts exhibited lower caspase-3 specific activity and activated hypoxia-inducible factor-1 protein , they had higher levels of mRNA for NF-κB , Bnip3 and caspase-1 mRNA . Increased 8-hydroxy-2'-deoxyguanosine levels reflected greater oxidative stress and reactive oxygen species-related DNA fragmentation . CONCLUSIONS : Our data indicate a significant role of pro-apoptotic and pro-inflammatory activity in allografts that subsequently exhibit primary graft failure . The relatively lower levels of apoptotic and inflammatory activity in P81605 hearts suggest they may represent a potentially usable donor cardiac allograft pool . This possibility requires further detailed molecular and clinical research . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . Role of NAD(P)H oxidase in transforming growth factor-beta1-induced monocyte chemoattractant protein-1 and interleukin-6 expression in rat renal tubular epithelial cells . AIM : This study investigated the role of NAD(P)H oxidase in transforming growth factor-beta1 ( TGF-beta1 ) -induced reactive oxygen species ( ROS ) generation , monocyte chemoattractant protein-1 ( P13500 ) , and interleukin-6 ( P05231 ) expression in rat renal tubular epithelial Q7Z2Y5 -52E cells . METHODS : The cells were treated with 10 ng/mL TGF-beta1 , either in the presence or absence of the NAD(P)H oxidase inhibitor , diphenyleneiodonium ( DPI ) , or short hairpin RNA ( shRNA ) suppressing p67phox expression . Expression of NAD(P)H oxidase subunits , P13500 , and P05231 at the mRNA levels was detected by reverse transcription polymerase chain reaction , while expression of NAD(P)H oxidase subunit p67phox protein was analyzed by western blot and P13500 by enzyme-linked immunosorbent assay . The cellular ROS generation was visualized using 2',7'-dichlorodihydrofluorescein diacetate by confocal microscopy . RESULTS : Compared to control , TGF-beta1 upregulated NAD(P)H oxidase subunit p67phox mRNA by 3.59-fold ( P < 0.01 ) , but had no effect on P13498 , gp91phox and p47phox NAD(P)H subunits . TGF-beta1 was also able to significantly increase intracellular ROS ( P < 0.05 ) , P13500 ( P < 0.01 ) and P05231 ( P < 0.05 ) expression in Q7Z2Y5 -52E cells . Further studies showed that generation of ROS and upregulation of P13500 and P05231 by TGF-beta1 were significantly blocked by addition of DPI or shRNA-p67phox ( P < 0.01 ) , suggesting that these effects were NAD(P)H oxidase-dependent . CONCLUSION : TGF-beta1 differentially regulates the expression of NAD(P)H oxidase subunits and mediates P13500 and P05231 expression in rat renal tubular cells via the NAD(P)H oxidase/p67phox-dependent mechanism . 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 . P38936 -activated kinase ( Pak ) regulates NADPH oxidase activation in human neutrophils . The phagocyte nicotinamide adenine dinucleotide phosphate ( NADPH ) oxidase plays an instrumental role in host defense and contributes to microbicial killing by releasing highly reactive oxygen species . This multicomponent enzyme is composed of membrane and cytosolic components that assemble in the plasma membrane or phagolysosome . While the guanosine S'-triphosphatase ( GTPase ) Rac2 has been shown to be a critical regulator of NADPH oxidase activity and assembly , the role of its effector , P38936 -activated kinase ( Pak ) , in oxidase function has not been well defined . Using HIV-1 Tat-mediated protein transduction of Pak inhibitory domain , we show here that Pak activity is indeed required for efficient superoxide generation in intact neutrophils . Furthermore , we show that Pak translocates to the plasma membrane upon N-formyl-methionyl-leucyl-phenylalanine ( fMLF ) stimulation and colocalizes with translocated p47(phox) and with P13498 , a subunit of flavocytochrome b558 . Although activated Pak phosphorylated several essential serine residues in the C-terminus of p47phox , direct binding to p47phox was not observed . In contrast , active Pak bound directly to P13498 , suggesting flavocytochrome b was the oxidase-associated membrane target of this kinase and this association may facilitate further phosphorylation of p47phox in the assembling NADPH oxidase complex . 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 . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . [ 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 . 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 . 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 . 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 . P01308 resistance determines phagocytic nicotinamide adenine dinucleotide phosphate oxidase overactivation in metabolic syndrome patients . OBJECTIVE : Metabolic syndrome ( MetS ) is associated with insulin resistance and increases the cardiovascular risk . Oxidative stress constitutes a potential mechanism that links insulin resistance and cardiovascular disease . The aim of this study was to analyze the relationship of NADPH oxidase activation with insulin resistance , and the effect of this interaction on the cardiovascular risk in MetS patients . METHODS : NADPH oxidase-dependent superoxide production and expression was evaluated by luminescence and western blot , respectively , in peripheral blood mononuclear cells obtained from 125 patients with MetS . P01308 resistance was defined by the homeostasis model assessment index . P14780 was quantified by enzyme-linked immunosorbent assay in plasma samples . To ascertain the mechanisms involved in vivo , we performed in-vitro experiments in cultured macrophages . RESULTS : Fifty-six percent of patients with MetS showed insulin resistance . Plasma matrix metalloproteinase-9 levels were higher ( P < 0.05 ) in insulin-resistant patients than in patients with insulin sensitivity . NADPH oxidase-dependent superoxide production was augmented ( P < 0.05 ) in insulin-resistant patients with respect to insulin-sensitive patients . The interaction between insulin resistance and abnormally high NADPH oxidase-mediated superoxide production was associated with the highest matrix metalloproteinase-9 values . Increased NADPH oxidase-dependent superoxide production was significantly associated with higher NADPH oxidase P13498 expression in insulin-resistant than in insulin-sensitive patients . Interestingly , insulin upregulated P13498 in peripheral blood mononuclear cells and in murine macrophages . CONCLUSION : P01308 resistance is associated with phagocytic NADPH oxidase activation . This association results in the highest cardiovascular risk in MetS patients . Cardioprotective mechanism of telmisartan via P37231 - P29474 pathway in dahl salt-sensitive hypertensive rats . BACKGROUND : Recently , some investigators have shown that telmisartan , an angiotensin II ( Ang II ) -receptor blocker ( ARB ) , is a partial agonist of the peroxisome proliferator-activated receptor-gamma ( P37231 ) . We investigate whether telmisartan improves cardiovascular remodeling associated with the production of endothelial nitric oxide synthase ( P29474 ) through P37231 , inhibits the Rho-kinase pathway , and suppresses oxidative stress in Dahl salt-sensitive ( DS ) hypertensive rats . METHODS : Telmisartan ( 1 mg/kg per day ) or telmisartan plus P37231 inhibitor , GW9662 ( 1 mg/kg per day ) was administered from the age of 6-11 weeks . Age-matched male Dahl salt-resistant ( DR ) rats served as a control group . RESULTS : The levels of P29474 and P37231 expression , and P29474 phosphorylation were significantly lower in DS rats than in DR rats . Chronic telmisartan treatment in DS rats significantly increased these parameters , but not telmisartan plus GW9662 . Telmisartan effectively inhibited the vascular lesion formation such as medial thickness and perivascular fibrosis , but not telmisartan plus GW9662 . Moreover , upregulated RhoA protein , Rho-kinase mRNA , and myosin light-chain phosphorylation in DS rats was decreased by telmisartan to a similar degree as observed after treatment with Y-27632 , a selective Rho-kinase inhibitor . In addition , NAD(P)H oxidase P13498 , p47phox , gp91phox expression , and mitogen-activated protein kinase and its downstream effector P08133 S6 kinase phosphorylation in DS rats was also inhibited by telmisartan . CONCLUSIONS : These results suggest that the cardioprotective mechanism of telmisartan may be partly due to improvement of endothelial function associated with P37231 - P29474 , oxidative stress , and Rho-kinase pathway . 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 . Changes in Dermatomal Somatosensory Evoked Potentials according to Stimulation Intensity and Severity of Carpal Tunnel Syndrome . OBJECTIVE : To investigate the change of latency of cervical dermatomal somatosensory evoked potential ( P81605 ) according to stimulation intensity ( SI ) and severity of carpal tunnel syndrome ( CTS ) . METHODS : Stimulation sites were the P13671 , P10643 , and Q99618 dermatomal areas . Two stimulation intensities 1.5×sensory threshold ( ST ) and 2.5×ST were used on both normal and CTS patients . RESULTS : In moderate CTS , the latencies of P13671 and P10643 P81605 during 1.5×ST SI and those of P10643 P81605 during 2.5×ST SI were significantly delayed compared with the values of normal subjects . Significant correlation between the latency of P10643 P81605 of 2.5×ST stimulation and the median sensory nerve conduction velocity was observed . CONCLUSION : We suggest that these data can aid in the diagnosis of cervical sensory radiculopathy using low stimulation intensity and of those who have cervical sensory radiculopathy combined with CTS patients . P33261 polymorphism is associated with reduced clopidogrel response in cerebrovascular disease . PURPOSE : DB00758 is a prodrug that requires transformation into an active metabolite by cytochrome P450 ( CYP ) in the liver in order to irreversibly inhibit the Q9H244 adenosine diphosphate platelet receptor . P33261 polymorphism has been reported to correlate with reduced antiplatelet activity of clopidogrel in coronary artery disease . We assessed the association between P33261 polymorphism and clopidogrel resistance in patients with cerebrovascular disease . MATERIALS AND METHODS : We retrospectively gathered data from patients who experienced cerebrovascular disease , received clopidogrel , and were tested for clopidogrel resistance and P33261 polymorphism . DB00758 resistance was tested by the VerifyNow Q9H244 system , and the P33261 polymorphism was tested by the Seeplex P33261 P12821 Genotyping system . DB00758 resistance was expressed in Q9H244 reaction units ( PRU ) and percent inhibition . High PRU and low percent inhibition suggests clopidogrel resistance . P33261 polymorphisms were expressed as extensive , intermediate , and poor metabolizers . DB00758 resistance was assessed according to the subgroup of P33261 polymorphism . RESULTS : A total of 166 patients were evaluated . The PRU values of extensive P33261 metabolizers ( 195.0±84.9 ) were significantly lower than those of intermediate and poor metabolizers ( 237.9±88.0 , 302.2±58.9 ) . The percent inhibition of extensive metabolizers ( 44.6±21.8 ) was significantly higher than that of intermediate and poor metabolizers ( 30.5±21.5 , 14.0±13.4 ) . CONCLUSION : Intermediate and poor metabolizing P33261 polymorphism is associated with reduced clopidogrel antiplatelet activity in patients with cerebrovascular disease . The clinical implications of this finding require further investigation . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 .