interacts_with DB00519?
Down-regulation of P12830 in mouse skin carcinoma cells enhances a migratory and invasive phenotype linked to matrix metalloproteinase-9 gelatinase expression . To assess the role of gelatinases in mouse skin tumor progression and their link to the expression of P12830 ( E-CD ) , the cell-cell adhesion protein , we used the highly metastatic squamous HaCa4 cell line and several HaCa4-derived clones obtained by transfection of the mouse E-CD cDNA . Expression of matrix metalloproteinase-9 ( P14780 ) mRNA and protein activity were present in E-CD ( - ) HaCa4 and control clones in culture , but they were strongly diminished in E-CD ( + ) clones ( E24 and E62 ) at subconfluence . To explore the suppressive effect of the cell-cell contacts mediated by E-CD on P14780 expression , we introduced a plasmid encoding mouse E-CD antisense cDNA into the E24 cell clone . The transfectant P1-clones obtained with reduced or absent E-CD expression showed increased levels of P14780 gelatinase , motility in vitro , and metastatic potential in vivo . Expression of P14780 in the various cell clones was also negatively modulated by cell density , but this effect was much stronger in E-CD ( + ) cells , despite the fact that all of the cell clones analyzed maintained the expression of P22223 and made cell-cell contacts at high cell density . Our results indicate that in this cell system , the E-CD-mediated cell-cell contacts are involved in the down-regulation of P14780 expression . Thus , the loss of E-CD triggers a migratory and invasive phenotype in mouse squamous carcinoma cells . 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 . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . 17β-estradiol inhibits P14780 and Q09428 /TrpM4 expression and activation and thereby attenuates BSCB disruption/hemorrhage after spinal cord injury in male rats . Blood-spinal cord barrier ( BSCB ) disruption and progressive hemorrhage after spinal cord injury ( SCI ) lead to secondary injury and the subsequent apoptosis and/or necrosis of neuron and glia , causing permanent neurological deficits . In this study , we examined the effect of 17β-estradiol ( E2 ) on BSCB breakdown and hemorrhage as well as subsequent inflammation after SCI . After a moderate contusion injury at the 9th thoracic segment of spinal cord , E2 ( 300 μg/kg ) was administered by iv injection immediately after SCI , and the same dose of E2 was then administered 6 and 24 hours after injury . Our data show that E2 attenuated BSCB permeability and hemorrhage and reduced the infiltration of neutrophils and macorphages after SCI . Consistent with this finding , the expression of inflammatory mediators was significantly reduced by E2 . Furthermore , E2 treatment significantly inhibited the expression of sulfonylurea receptor 1 and transient receptor potential melastatin 4 after injury , which are known to mediate hemorrhage at an early stage after SCI . Moreover , the expression and activation of matrix metalloprotease-9 after injury , which is known to disrupt BSCB , and the degradation of tight junction proteins , such as zona occludens-1 and occludin , were significantly inhibited by E2 treatment . Furthermore , the protective effects of E2 on BSCB disruption and functional improvement were abolished by an estrogen receptor antagonist , ICI 182780 ( 3 mg/kg ) . Thus , our study provides evidence that the neuroprotective effect of E2 after SCI is , in part , mediated by inhibiting BSCB disruption and hemorrhage through the down-regulation of sulfonylurea receptor 1/transient receptor potential melastatin 4 and matrix metalloprotease-9 , which is dependent on estrogen receptor . 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 . [ Network pharmacology study on major active compounds of Fufang Danshen formula ] . OBJECTIVE : To investigate the correlations between multi-compounds of Fufang Danshen formula and their multi targets and multi diseases . METHOD : Literature knowledge of nine major active compounds from Fufang Danshen formula , including tanshinone II(A) , salvianolic acid B , protocatechuic aldehyde , danshensu , cryptotanshinone , notoginsenoside Q96GN5 , ginsenoside Rg1 , DB06749 and borneol were collected from PubMed . Combined with cardiovascular related diseases and genes from OMIM database , the corresponding multi-compound-multi- target-multi-disease network was constructed and visualized by Cytoscape software . RESULT : AND CONCLUSION : Network analysis showed that the 9 compounds could modulate 42 cardiovascular associated genes ( e. g. P37231 , P12821 , Q14654 , P51787 , Q09428 , et al ) , which related to 30 cardiovascular associated diseases including non-insulin-dependent diabetes mellitus , hyperinsulinemic hypoglycemia , hypertension , and coronary heart disease . These results suggested new potential indications of Fufang Danshen formula . [ 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 . 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 . A New Histone Deacetylase Inhibitor , MHY219 , Inhibits the Migration of Human Prostate Cancer Cells via Q13547 . Histone deacetylase ( HDAC ) inhibitors are considered novel agents for cancer chemotherapy . We previously investigated MHY219 , a new HDAC inhibitor , and its potent anticancer activity in human prostate cancer cells . In the present study , we evaluated MHY219 molecular mechanisms involved in the regulation of prostate cancer cell migration . Similar to suberanilohydroxamic acid ( DB02546 ) , MHY219 inhibited Q13547 enzyme activity in a dose-dependent manner . MHY219 cytotoxicity was higher in LNCaP ( IC50=0.67 μM ) than in DU145 cells ( IC50=1.10 μM ) and PC3 cells ( IC50=5.60 μM ) after 48 h of treatment . MHY219 significantly inhibited the Q13547 protein levels in LNCaP and DU145 cells at high concentrations . However , inhibitory effects of MHY219 on HDAC proteins levels varied based on the cell type . MHY219 significantly inhibited LNCaP and DU145 cells migration by down-regulation of matrix metalloprotease-1 ( P03956 ) and P08253 and induction of tissue inhibitor of metalloproteinases-1 ( P01033 ) . These results suggest that MHY219 may potentially be used as an anticancer agent to block cancer cell migration through the repression of P03956 and P08253 , which is related to the reduction of Q13547 . Identification and characterization of nucleolin as a COUP-TFII coactivator of retinoic acid receptor β transcription in breast cancer cells . INTRODUCTION : The orphan nuclear receptor COUP-TFII plays an undefined role in breast cancer . Previously we reported lower COUP-TFII expression in tamoxifen/endocrine-resistant versus sensitive breast cancer cell lines . The identification of COUP-TFII-interacting proteins will help to elucidate its mechanism of action as a transcriptional regulator in breast cancer . RESULTS : FLAG-affinity purification and multidimensional protein identification technology ( MudPIT ) identified nucleolin among the proteins interacting with COUP-TFII in MCF-7 tamoxifen-sensitive breast cancer cells . Interaction of COUP-TFII and nucleolin was confirmed by coimmunoprecipitation of endogenous proteins in MCF-7 and T47D breast cancer cells . In vitro studies revealed that COUP-TFII interacts with the C-terminal arginine-glycine repeat ( RGG ) domain of nucleolin . Functional interaction between COUP-TFII and nucleolin was indicated by studies showing that siRNA knockdown of nucleolin and an oligonucleotide aptamer that targets nucleolin , DB04998 , inhibited endogenous COUP-TFII-stimulated RARB2 expression in MCF-7 and T47D cells . Chromatin immunoprecipitation revealed COUP-TFII occupancy of the RARB2 promoter was increased by all-trans retinoic acid ( atRA ) . RARβ2 regulated gene RRIG1 was increased by atRA and COUP-TFII transfection and inhibited by siCOUP-TFII . Immunohistochemical staining of breast tumor microarrays showed nuclear COUP-TFII and nucleolin staining was correlated in invasive ductal carcinomas . COUP-TFII staining correlated with ERα , Q15788 , Q9Y6Q9 , Pea3 , P08253 , and phospho-Src and was reduced with increased tumor grade . CONCLUSIONS : Our data indicate that nucleolin plays a coregulatory role in transcriptional regulation of the tumor suppressor RARB2 by COUP-TFII . P62158 interacts with angiotensin-converting enzyme-2 ( Q9BYF1 ) and inhibits shedding of its ectodomain . P12821 -2 ( Q9BYF1 ) is a regulatory protein of the renin-angiotensin system ( DB01367 ) and a receptor for the causative agent of severe-acute respiratory syndrome ( P49591 ) , the P49591 -coronavirus . We have previously shown that Q9BYF1 can be shed from the cell surface in response to phorbol esters by a process involving P01375 converting enzyme ( P78536 ; P78536 ) . In this study , we demonstrate that inhibitors of calmodulin also stimulate shedding of the Q9BYF1 ectodomain , a process at least partially mediated by a metalloproteinase . We also show that calmodulin associates with Q9BYF1 and that this interaction is decreased by calmodulin inhibitors . 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 . 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 . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Is transforming growth factor-β signaling activated in human hypertrophied prostate treated by 5-alpha reductase inhibitor ? BACKGROUND AND AIM : It is well known that androgen deprivation relates to penile fibrosis , so we hypothesize that long-term treatment with 5-alphareductase inhibitors ( 5ARIs ) may increase the risk of fibrosis of prostate . PATIENTS AND METHODS : Thirty-two BPH patients who underwent transurethral resection of the prostate were enrolled . The patients were divided into two groups : group one , 16 patients underwent TURP who had been treated with tamsulosin for 2 years ; group two , 16 patients underwent TURP who had been treated with combination of tamsulosin and dutasteride for at least 1 year . We evaluated the expressions of P29475 , P35228 , P29474 , TGF-β1 , TGF-β2 , phosphorylated- Q15796 /3 ( p- Q15796 /3 ) , P12830 , P19022 , and α-smooth muscle actin in the resected prostate tissues by western blotting , and the TGF-β concentration was determined by ELISA kit . RESULTS : The expressions of 3 isoforms of NOS were significantly increased in group 2 except of P29474 in lateral prostate , and the expressions of TGF-β1 , TGF-β2 , and p- Q15796 /3 increased about 2-fold compared with group 1 . In group 2 , the P12830 expression decreased while P19022 expression increased significantly . CONCLUSIONS : The overexpression of P29475 may contribute to prostate smooth muscle relaxation ; however , long-time treatment with 5 Q9Y4X5 increases the risk of fibrosis of prostate . 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 . Inhibition of cellular growth and migration by suppression of endothelial protein C receptor ( Q9UNN8 ) in lung carcinoma cells . Increasing evidence shows that beyond its role in coagulation , thrombosis interferes with carcinogenesis . Q9UNN8 ( Q9UNN8 ) is a cellular receptor for protein C and activated protein C ( P25054 ) . Such Q9UNN8 -induced signal transduction promotes cancer cell migration , invasion , and angiogenesis and inhibits cancer cell apoptosis . However , its role in lung carcinoma biology is yet to be demonstrated . Here , the recombinant Q9UNN8 siRNA plasmids were constructed to investigate the effects of inhibition of Q9UNN8 on human lung cancer H1299 . Q9UNN8 siRNA led to inhibition of endogenous Q9UNN8 mRNA and protein expression as determined by RT-PCR and Western blotting analysis . Q9UNN8 siRNA significantly inhibited cell growth , blocked entry into the S phase of the cell cycle , and reduced the migration of H1299 cells . Q9UNN8 siRNA also decreases P08253 and cyclin E expression in H1299 cells . In addition , siRNA targeting of Q9UNN8 inhibited the growth of H1299 cells and decreased P53602 in SCID mice tumor models . Taken together , Q9UNN8 was involved in regulating progression of human lung cancer cells . Manipulation of Q9UNN8 expression may be a potential therapeutic strategy for lung cancer . Selective inhibition of histone deacetylase 6 ( Q9UBN7 ) induces DNA damage and sensitizes transformed cells to anticancer agents . Q9UBN7 ( Q9UBN7 ) is structurally and functionally unique among the 11 human zinc-dependent histone deacetylases . Here we show that chemical inhibition with the Q9UBN7 -selective inhibitor tubacin significantly enhances cell death induced by the topoisomerase II inhibitors etoposide and doxorubicin and the pan-HDAC inhibitor DB02546 ( vorinostat ) in transformed cells ( LNCaP , MCF-7 ) , an effect not observed in normal cells ( human foreskin fibroblast cells ) . The inactive analogue of tubacin , nil-tubacin , does not sensitize transformed cells to these anticancer agents . Further , we show that down-regulation of Q9UBN7 expression by shRNA in LNCaP cells enhances cell death induced by etoposide , doxorubicin , and DB02546 . Tubacin in combination with DB02546 or etoposide is more potent than either drug alone in activating the intrinsic apoptotic pathway in transformed cells , as evidenced by an increase in PARP cleavage and partial inhibition of this effect by the pan-caspase inhibitor Z-VAD-fmk . Q9UBN7 inhibition with tubacin induces the accumulation of γ P16104 , an early marker of DNA double-strand breaks . Tubacin enhances DNA damage induced by etoposide or DB02546 as indicated by increased accumulation of γ P16104 and activation of the checkpoint kinase Chk2 . Tubacin induces the expression of P35638 ( P35638 / P35638 ) , a transcription factor up-regulated in response to cellular stress . P35638 induction is further increased when tubacin is combined with DB02546 . These findings point to mechanisms by which Q9UBN7 -selective inhibition can enhance the efficacy of certain anti-cancer agents in transformed cells . 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 . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Arm-cranking exercise assisted by Functional Electrical Stimulation in P13671 tetraplegia : a pilot study . Tetraplegic volunteers undertook progressive exercise training , using novel systems for arm-cranking exercise assisted by Functional Electrical Stimulation ( P07332 ) . The main aim was to determine potential training effects of P07332 -assisted arm-crank ergometry ( P07332 - P12821 ) on upper limb strength and cardiopulmonary ( fitness ) in tetraplegia . Surface P07332 was applied to the biceps and triceps during exercise on an instrumented ergometer . Two tetraplegic volunteers with P13671 Spinal Cord Injury ( SCI ) went through muscle strengthening , baseline exercise testing and three months of progressive P07332 - P12821 training . Repeat exercise tests were carried out every four weeks during training , and post-training , to monitor upper-limb strength and cardiopulmonary fitness . At each test point , an incremental test was carried out to determine peak work rate , peak oxygen uptake , gas exchange threshold and oxygen uptake-work rate relationship during P07332 - P12821 . Peak oxygen uptake for Subject A increased from 0.7 l/min to 1.1 l/min , and peak power output increased from 7 W to 38 W after P07332 - P12821 training . For Subject B , peak oxygen uptake was unchanged , but peak power output increased from 3 W to 8 W . These case studies illustrate potential benefits of P07332 - P12821 in tetraplegia , but also the differences in exercise responses between individuals . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . 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 . The sulphydryl containing P12821 inhibitor Zofenoprilat protects coronary endothelium from Doxorubicin-induced apoptosis . Pediatric and adult cancer patients , following the use of the antitumor drug Doxorubicin develop cardiotoxicity . Pharmacological protection of microvascular endothelium might produce a double benefit : ( i ) reduction of myocardial toxicity ( the primary target of Doxorubicin action ) and ( ii ) maintenance of the vascular functionality for the adequate delivery of chemotherapeutics to tumor cells . This study was aimed to evaluate the mechanisms responsible of the protective effects of the angiotensin converting enzyme inhibitor ( ACEI ) Zofenoprilat against the toxic effects exerted by Doxorubicin on coronary microvascular endothelium . We found that exposure of endothelial cells to Doxorubicin ( 0.1-1μM range ) impaired cell survival by promoting their apoptosis . P27361 /2 related p53 activation , but not reactive oxygen species , was responsible for Doxorubicin induced caspase-3 cleavage . P04637 mediated-apoptosis and impairment of survival were reverted by treatment with Zofenoprilat . The previously described PI-3K/ P29474 /endogenous fibroblast growth factor signaling was not involved in the protective effect , which , instead , could be ascribed to cystathionine gamma lyase dependent availability of H2S from Zofenoprilat . Furthermore , considering the tumor environment , the treatment of endothelial/tumor co-cultures with Zofenoprilat did not affect the antitumor efficacy of Doxorubicin . In conclusion the ACEI Zofenoprilat exerts a protective effect on Doxorubicin induced endothelial damage , without affecting its antitumor efficacy . Thus , sulfhydryl containing ACEI may be a useful therapy for Doxorubicin-induced cardiotoxicity . Thrombin has biphasic effects on the nitric oxide-cGMP pathway in endothelial cells and contributes to experimental pulmonary hypertension . BACKGROUND : A potential role for coagulation factors in pulmonary arterial hypertension has been recently described , but the mechanism of action is currently not known . Here , we investigated the interactions between thrombin and the nitric oxide-cGMP pathway in pulmonary endothelial cells and experimental pulmonary hypertension . PRINCIPAL FINDINGS : Chronic treatment with the selective thrombin inhibitor melagatran ( 0.9 mg/kg daily via implanted minipumps ) reduced right ventricular hypertrophy in the rat monocrotaline model of experimental pulmonary hypertension . In vitro , thrombin was found to have biphasic effects on key regulators of the nitric oxide-cGMP pathway in endothelial cells ( HUVECs ) . Acute thrombin stimulation led to increased expression of the cGMP-elevating factors endothelial nitric oxide synthase ( P29474 ) and soluble guanylate cyclase ( sGC ) subunits , leading to increased cGMP levels . By contrast , prolonged exposition of pulmonary endothelial cells to thrombin revealed a characteristic pattern of differential expression of the key regulators of the nitric oxide-cGMP pathway , in which specifically the factors contributing to cGMP elevation ( P29474 and sGC ) were reduced and the cGMP-hydrolyzing O76074 was elevated ( qPCR and Western blot ) . In line with the differential expression of key regulators of the nitric oxide-cGMP pathway , a reduction of cGMP by prolonged thrombin stimulation was found . The effects of prolonged thrombin exposure were confirmed in endothelial cells of pulmonary origin ( HPAECs and HPMECs ) . Similar effects could be induced by activation of protease-activated receptor-1 ( P25116 ) . CONCLUSION : These findings suggest a link between thrombin generation and cGMP depletion in lung endothelial cells through negative regulation of the nitric oxide-cGMP pathway , possibly mediated via P25116 , which could be of relevance in pulmonary arterial hypertension . Protective effects of heparin on endothelial cells in sepsis . This study aims to observe the protective effects of heparin on endothelial cells in sepsis and explore the involved signal pathway regulated by heparin . Methods Human vascular endothelial cells were treated by TNFα in vitro to simulate the inflammatory environment when sepsis occurred . They were intervened by heparin and the expression levels of soluble thrombomodulin ( sTM ) and serum activated protein C ( P25054 ) were detected by ELISA , the regulatory mechanism of heparin improving vascular endothelial cells injury induced by TNFα was detected by Western Blotting method , the methylation of histone in the gene promoter region of endothelial nitric oxide synthase ( P29474 ) and monocyte chemotactic protein-1 ( P13500 ) were detected using chromatin immunoprecipitation method . Results DB01109 could inhibit the secretion of sTM and P25054 protein and the expression of P13500 gene which involved in NF-κB signal pathway . Conclusions DB01109 could protect vascular endothelial cells from injury induced by TNFα and sepsis , the mechanisms were related with the effects of heparin on the histone methylation of promoter region and the regulation of heparin on the MAPK and NF-κB signal pathways . These results provide a theoretical basis for the application of heparin in the prevention and treatment of vascular disease related with sepsis . 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) . Role of genetic polymorphisms of ion channels in the pathophysiology of coronary microvascular dysfunction and ischemic heart disease . Conventionally , ischemic heart disease ( IHD ) is equated with large vessel coronary disease . However , recent evidence has suggested a role of compromised microvascular regulation in the etiology of IHD . Because regulation of coronary blood flow likely involves activity of specific ion channels , and key factors involved in endothelium-dependent dilation , we proposed that genetic anomalies of ion channels or specific endothelial regulators may underlie coronary microvascular disease . We aimed to evaluate the clinical impact of single-nucleotide polymorphisms in genes encoding for ion channels expressed in the coronary vasculature and the possible correlation with IHD resulting from microvascular dysfunction . 242 consecutive patients who were candidates for coronary angiography were enrolled . A prospective , observational , single-center study was conducted , analyzing genetic polymorphisms relative to ( 1 ) NOS3 encoding for endothelial nitric oxide synthase ( P29474 ) ; ( 2 ) P16615 encoding for the Ca²⁺/H⁺-ATPase pump ( SERCA ) ; ( 3 ) Q14524 encoding for the voltage-dependent Na⁺ channel ( Nav1.5 ) ; ( 4 ) Q15842 and Q14654 encoding for the Kir6.1 and Kir6.2 subunits of K- DB00171 channels , respectively ; and ( 5 ) KCN5A encoding for the voltage-gated K⁺ channel ( Kv1.5 ) . No significant associations between clinical IHD manifestations and polymorphisms for SERCA , Kir6.1 , and Kv1.5 were observed ( p > 0.05 ) , whereas specific polymorphisms detected in P29474 , as well as in Kir6.2 and Nav1.5 were found to be correlated with IHD and microvascular dysfunction . Interestingly , genetic polymorphisms for ion channels seem to have an important clinical impact influencing the susceptibility for microvascular dysfunction and IHD , independent of the presence of classic cardiovascular risk factors . DB04540 attenuates linoleic acid-induced endothelial cell activation . Vascular endothelial cell activation and dysfunction are critical early events in atherosclerosis . Even though very low or high levels of cholesterol can compromise cellular functions , cholesterol is a critical membrane component and may protect the vascular endothelium from oxidative stress and polyunsaturated fatty acid-mediated inflammatory responses . We have previously shown that the parent omega-6 fatty acid linoleic acid can markedly activate vascular endothelial cells . We now propose that membrane cholesterol can modify and inhibit linoleic acid-mediated endothelial cell dysfunction . To test this hypothesis , pulmonary artery endothelial cells were incubated with cholesterol ( 0 to 100 micromol/L ) for 24 hours and then treated with 90 micromol/L of linoleic acid ( 18:2n-6 ) for 6 to 24 hours . In control cells , treatment with linoleic acid reduced intracellular glutathione levels and induced the DNA binding activity of nuclear factor-kappaB ( NF-kappaB ) leading to the upregulation of interleukin-6 ( P05231 ) . In addition , the expression of endothelial nitric oxide synthase ( P29474 ) was altered , with linoleic acid increasing P29474 activity . In contrast , enrichment with cholesterol enhanced glutathione levels and reduced the linoleic acid-induced activation of NF-kappaBand the production of P05231 . Prior exposure to 50 micromol/L cholesterol also prevented the fatty acid-induced increase in P29474 activation . DB04540 loading activated peroxisome proliferator-activated receptor-gamma ( P37231 ) , a nuclear receptor that can decrease inflammatory responses . Furthermore , the P37231 agonist thiazolidinedione markedly downregulated the NF-kappaB activation mediated by linoleic acid . Our data suggest that signaling pathways linked to endothelial cell activation by prooxidant and proinflammatory insults may be influenced by cellular cholesterol levels . Apigenin up-regulates transgelin and inhibits invasion and migration of colorectal cancer through decreased phosphorylation of AKT . Colorectal cancer ( CRC ) is a major cause of morbidity and mortality throughout the world . Apigenin is a flavonoid that possesses various clinically relevant properties such as anti-tumour , anti-platelet and anti-inflammatory activities . Our results showed that apigenin has anti-proliferation , anti-invasion and anti-migration effects in three kinds of colorectal adenocarcinoma cell lines , namely SW480 , DLD-1 and LS174T . Proteomic analysis with SW480 indicated that apigenin up-regulated the expression of transgelin ( Q01995 ) in mitochondria to exert its anti-tumour growth and anti-metastasis effects . Real-time quantitative polymerase chain reaction ( RQ-PCR ) and western blot confirm the up-regulation in all the three colorectal adenocarcinoma cells . An inverse correlation was observed between Q01995 expression and CRC metastasis in tissue microarray staining . Q01995 siRNA increased the viability of SW480 . Apigenin decreased the expression of P14780 in a dose-dependent manner . Transfection of three truncated forms of Q01995 and wild type has identified Q01995 as a repressor of P14780 expression . A synergetic effect was observed in overexpression of Q01995 wild type and apigenin treatment which manifested as lowered phosphorylation of AKT Ser473 and Q06187 Thr308 . In an orthotopic CRC model , apigenin inhibited tumour growth and metastasis to liver and lung . In conclusion , our research provided direct evidence that apigenin inhibited tumour growth and metastasis both in vitro and in vivo . Apigenin up-regulated Q01995 and hence down-regulated P14780 expression through decreasing phosphorylation of Akt at Ser473 and in particular Thr308 to prevent cell proliferation and migration . Rationale for double renin-angiotensin-aldosterone system blockade . The clinical benefits of both angiotensin-converting enzyme ( P12821 ) inhibitors and angiotensin receptor blockers ( ARBs ) extend beyond blood pressure reduction to encompass tissue-protective effects in target organs , such as the heart , vasculature , and kidneys , that underlie the reductions in cardiovascular mortality and morbidity seen in large outcome trials . However , these effects are achieved by different mechanisms . P12821 inhibitors reduce circulating and tissue angiotensin II levels and potentiate the beneficial effects of bradykinin , including generation of nitric oxide ( NO ) . By contrast , the protective effects of ARBs are owing to the blockade of the angiotensin II type 1 ( AT(1) ) receptors and possibly also to the stimulation of angiotensin II type 2 ( AT(2) ) receptors , again resulting in NO release . In addition , some ARBs , such as telmisartan , are selective activators of peroxisome proliferator-activated receptor-gamma ( P37231 ) , thereby increasing insulin sensitivity . In contrast to other P37231 ligands , such as the thiazolidinediones , activation of this receptor by telmisartan does not result in weight gain . The complementary mechanisms of action of P12821 inhibitors and ARBs create a rationale for combination therapy in high-risk patients . The benefits of this approach with telmisartan are being investigated in clinical trials , such as the Ongoing Telmisartan Alone in Combination with Ramipril Global Endpoint Trial ( ONTARGET ) . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . 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 . P00797 status does not predict the anti-hypertensive response to angiotensin-converting enzyme inhibition in African-Americans . DB00519 Multicenter Study Group . The angiotensin-converting enzyme ( P12821 ) inhibitor trandolapril , a non-sulfhydryl prodrug which is hydrolysed into trandolaprilat , was studied in 322 hypertensives of African-American descent using a double-blind , randomised , placebo-controlled , parallel study design . Following 6 weeks of double-blind treatment with placebo or 0.25 to 16 mg/day trandolapril , an analysis of drug effect on trough blood pressure ( BP ) stratified by age , gender , weight , pre-treatment plasma renin activity , and trandolaprilat concentration was performed . Two mg was the lowest effective trandolapril dose , whereas doses above 4 mg did not significantly reduce trough BP . Reduction in BP did not correlate with trough plasma trandolaprilat concentration . Pre-treatment plasma renin activity was not a reliable indicator of anti-hypertensive response , as similar reductions in BP occurred even in patients with the lowest renin levels . There were no observable differences based on age , gender or measurements of the renin-angiotensin-aldosterone axis . In conclusion , neither age , gender or plasma renin activity influenced anti-hypertensive response to angiotensin-converting enzyme inhibition in African-Americans . Gene polymorphisms impact the risk of rejection with hemodynamic compromise : a multicenter study . BACKGROUND : Rejection with hemodynamic compromise ( P18577 ) is associated with high mortality in heart recipients . This study investigates the association between genetic polymorphisms and P18577 in pediatric heart recipients . METHODS : Data from 532 pediatric heart recipients from six centers in the Pediatric Heart Transplant Study were analyzed for time to P18577 by recipient race , age at transplantation , and genotype at 13 genetic polymorphisms ( P01375 -α A-308G , P05231 G-174C , P27352 -γ T+874A , P22301 G-1082A , C-819T , and C-592A ; FAS A-670G , P48023 C-843T , and P12821 I/D ; and P15692 A-2578C , C-1451T , C+405G , and -2549 I/D ) . RESULTS : P18577 occurred in 126 ( 23.7 % ) patients during the study period . Adjusting for age and race , P22301 G-1082A , FAS A-670G , and P12821 I/D genotypes were associated with P18577 . P22301 G-1082A GG genotype was associated with decreased risk of P18577 with an adjusted hazard ratio ( HR ) of 0.49 ( 95 % confidence interval [ CI ] , 0.27-0.90 ; P=0.020 ) . FAS A-670G AA genotype was associated with increased risk of P18577 with an adjusted HR of 1.84 ( 95 % CI , 1.25-2.69 ; P=0.002 ) . P12821 II genotype was associated with decreased risk of P18577 with an adjusted HR of 0.58 ( 95 % CI , 0.36-0.95 ; P=0.031 ) . CONCLUSIONS : Recipients with a high anti-inflammatory and immune-regulatory genetic profile ( high interleukin-10 ) were protected from P18577 . Conversely , recipients with a pro-apoptotic genetic profile ( high Fas ) or high angiotensin-1-converting enzyme producing genotype were at increased risk of P18577 . This represents progress toward understanding the genetic risk factors of posttransplantation outcomes in pediatric heart recipients . Inflammation-mediated abrogation of androgen signaling : an in vitro model of prostate cell inflammation . As a link between inflammation and cancer has been reported in many studies , we established an in vitro model of prostatic inflammation to investigate the loss of androgen receptor ( AR ) -mediated signaling in androgen responsive prostate cell lines . First , the U937 monocyte cell line was differentiated into macrophages using phorbol acetate ( PMA ) , and cells were induced with lipopolysaccharide ( LPS ) for cytokine secretion . Next , the cytokine levels ( P01375 α , P05231 , and IL1β ) in conditioned media ( CM ) were analyzed . Prostate cells were then fed with CM containing varying concentrations of P01375 α , and IkB degradation , nuclear factor kappa B ( NFκB ) translocation and transactivation , and the expression of matrix metalloproteinase-8 ( P22894 ) and matrix metalloproteinase-9 ( P14780 ) were then assessed . As a result of CM treatment , ubiquitin-mediated AR degradation , which was restored using anti- P01375 α antibody neutralization , led to both a decrease in Q9Y5K2 , PSA , and Q99801 expression levels and the upregulation of P18283 . In addition to the loss of AR , acute and chronic CM exposure resulted in p53 degradation and consequent P38936 downregulation , which was also restored by either androgen administration or ectopic Q99801 expression via the stabilization of Q00987 levels in LNCaP cells . Additionally , CM treatment enhanced P16104 ( ( S139 ) ) phosphorylation ( a hallmark of DNA damage ) and genetic heterogeneity in the absence of androgens in prostate cells without activating mitochondrial apoptosis . Thus , the data suggest that inflammatory cytokine exposure results in the loss of AR and p53 signaling in prostate cells and facilitates genetic heterogeneity via ROS accumulation to promote prostate carcinogenesis . [ 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 . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . DEF6 , a novel substrate for the Tec kinase Q08881 , contains a glutamine-rich aggregation-prone region and forms cytoplasmic granules that co-localize with P-bodies . Localization of DEF6 ( SLAT/ Q9H4E7 ) , a Rho-family guanine nucleotide exchange factor , to the center of the immune synapse is dependent upon Q08881 , a Tec-family kinase that regulates the spatiotemporal organization of components of T cell signaling pathways and Cdc42-dependent actin polymerization . Here we demonstrate that Q08881 both interacts with DEF6 and phosphorylates DEF6 at tyrosine residues DB00135 (210) and DB00135 (222) . Expression of a GFP-tagged Y210E-Y222E phosphomimic resulted in the formation of DEF6 cytoplasmic granules that co-localized with decapping enzyme 1 ( P12821 ) , a marker of P-bodies ; sites of mRNA degradation . Similarly treatment of cells with puromycin or sodium arsenite , reagents that arrest translation , also resulted in the accumulation of DEF6 in cytoplasmic granules . Bioinformatics analysis identified a glutamine-rich , heptad-repeat region ; a feature of aggregating proteins , within the C-terminal region of DEF6 with the potential to promote granule formation through a phosphorylation-dependent unmasking of this region . These data suggest that in addition to its role as a Q9NR83 , DEF6 may also function in regulating mRNA translation . Endothelial dysfunction in congestive heart failure : P12821 inhibition vs. angiotensin II antagonism . BACKGROUND : Endothelial dysfunction of the vasculature contributes to the elevated peripheral resistance and reduced myocardial perfusion in congestive heart failure ( CHF ) . The present study systematically investigated the effect of angiotensin II ( AT(1) ) - receptor blockade on vascular superoxide ( O(2)(-) ) production and endothelial dysfunction . METHODS AND RESULTS : Vasodilator responses and O(2)(-) production were determined in aortic rings from Wistar rats with experimental CHF 10 weeks after extensive myocardial infarction and compared with sham-operated animals ( Sham ) . Rats were either treated with placebo ( P ) , with the AT(1)-receptor antagonist Irbesartan ( 50 mg kg(-1) day(-1) ) or with the P12821 inhibitor DB00519 ( 0.3 mg kg(-1) day(-1) ) . In CHF-P , endothelium-dependent , acetylcholine-induced relaxation was significantly attenuated compared with Sham-P . Chronic treatment with DB00519 or Irbesartan significantly improved endothelium-dependent relaxation . Aortic O(2)(-) formation was markedly increased in CHF , and was not significantly affected by DB00519 treatment , while it was reduced by Irbesartan. P29474 expression was reduced in CHF and normalised by both treatments . CONCLUSION : Endothelial vasomotor function in CHF rats was normalised by long-term treatment with an P12821 inhibitor or an AT(1)-antagonist . Reduced aortic P29474 expression was normalised by both treatments , whereas aortic superoxide formation was only reduced by the AT(1)-antagonist Irbesartan . Cytoskeletal changes during epithelial-to-fibroblastoid conversion as a crucial mechanism of des-gamma-carboxy prothrombin production in hepatocellular carcinoma . Des-gamma-carboxy prothrombin ( P12821 ) is a well-established tumor marker for hepatocellular carcinoma ( HCC ) , but the precise mechanism by which HCC cells produce P12821 remains unknown . Our preliminary experiments demonstrated that HepG2 cells with chemical induction of epithelial-to-fibroblastoid conversion ( EFC ) produced P12821 through impairment of vitamin K uptake via cytoskeletal rearrangement . Therefore , in this study we further examined this mechanism in vitro and using human HCC samples . Hepatoma cell lines ( HepG2 and P98160 /PRF/5 ) were induced EFC or epithelial-mesenchymal transition ( EMT ) by phorbol 12-myristate 13 acetate ( TPA ) or transforming growth factor ( TGF ) -beta1 . We analyzed these cells by ELISA , Western blotting and immunofluorescent studies . We also examined P12821 production and P12830 expression in human surgically resected HCC samples by immunohistochemical studies . Labeled low-density lipoprotein ( LDL ) uptake ( as a surrogate for vitamin K ) was significantly impaired in P12821 -producing hepatoma cells following induction of EFC or EMT . Further , filamentous actin , which plays a critical role in clathrin-mediated endocytosis , was dissociated in P12821 -producing cells . DB02621 , an actin depolymerizer , induced naïve hepatoma cells to produce P12821 with impairment of labeled-LDL uptake . In addition , an P12830 neutralizing antibody did not induce P12821 production . Finally , immunohistochemical studies demonstrated that P12821 production was inversely correlated with the intensity of P12830 expression in human HCC cells . In conclusion , cytoskeletal changes during EFC or EMT plays a critical mechanistic role in P12821 production via impairments in vitamin K uptake . 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 ) . Inhibition of matrix metalloproteinase-9 activity by trandolapril after middle cerebral artery occlusion in rats . We investigated whether an angiotensin-converting enzyme ( P12821 ) inhibitor could inhibit matrix metalloproteinase ( MMP ) activities in cerebral infarct lesions after middle cerebral artery occlusion ( MCAO ) in rats . After placebo or trandolapril ( 5 mg/kg per day ) was administered orally for 7 days , we permanently occluded the right middle cerebral artery . P12821 activity in extracts from the infarct side of placebo-treated rats was significantly higher than that in extracts from the non-infarct side from 5 days after MCAO , though they did not differ at 1 day . P12821 activities in extracts from both hemispheric segments in the trandolapril-treated group were significantly decreased compared with those in the placebo-treated group before MCAO , and this significant reduction persisted even at 7 days after MCAO . In the placebo-treated group , P14780 and P08253 activities in the infarct side were significantly increased at 12 h and at 1 day after MCAO , respectively . DB00519 treatment significantly reduced P14780 and P08253 activities to 68.5 % and 53.2 % , respectively . Seven days after MCAO , the ratios of infarct areas to the hemispheric sectional areas in placebo- and trandolapril-treated rats were 55.4+/-2.1 % and 30.9+/-2.9 % , respectively , and this difference was significant . Neurological severity scores were significantly improved from 1 to 7 days after MCAO in trandolapril-treated rats . Cumulative survival in trandolapril-treated rats was significantly increased compared with that in placebo-treated rats . Thus , the inhibition of P14780 by trandolapril might be part of the mechanism that prevents cerebral damage after cerebral ischemia . 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside ameliorates vascular senescence and improves blood flow involving a mechanism of p53 deacetylation . BACKGROUND AND AIMS : 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside ( THSG ) , a resveratrol analog with glucoside , has been shown in various studies to inhibit proliferation of vascular smooth muscle cells , attenuate inflammation , and prevent vascular endothelial dysfunction . In the study , we examined the effects of THSG on vascular senescence and blood flow . METHODS AND RESULTS : Oral administration of THSG for 14 weeks , resulted in notable increases in blood flow in spontaneously hypertensive rats ( SHRs ) ; and effective inhibition of vascular senescence as indicated by senescence-associated β-galactosidase ( SA-β-gal ) staining , phosphorylation of γ P16104 observed by stain analysis of immunofluorescence , and K373 acetylation of p53 in the aortic arches of SHRs . Oral administration of THSG also induced P29474 expression and urinary NOx production . THSG weekly activated Q96EB6 activity , stimulated P29474 promoter reporter gene activity , and ameliorated H(2)O(2)-induced cellular senescence and K373 acetylation of p53 in cultured human umbilical vein endothelial cells ( HUVECs ) . CONCLUSIONS : THSG improves blood flow and ameliorates vascular senescence by increasing P29474 expression and Sirt1 activity and decreasing acetylation of p53 at K373 site , at least in part , both in vitro and in vivo . Differential induction of matrix metalloproteinase 1 and 2 in ectopic endometrium . According to the transplantation theory , endometriosis develops from endometrial fragments that are retrogradely menstruated into the peritoneal cavity . In order to develop into endometriotic lesions , they have to connect to the vascular system by angiogenesis , probably involving matrix metalloproteinases ( MMP ) as key enzymes in extracellular matrix remodelling . A model of endometriosis using the chorioallantoic membrane ( P62158 ) of chick embryos was established . Eutopic endometrium from healthy women was transferred to the P62158 and cultivated ectopically for up to 3 days . Before transplantation and after 24 , 48 and 72 h of culture on the P62158 , total RNA was extracted and reverse transcribed . Human P03956 ( interstitial collagenase ) and P08253 ( gelatinase A ) mRNA expression was assessed by competitive PCR . Results were normalized to the content of human glyceraldehyde 3-phosphate dehydrogenase ( P04406 ) mRNA . In eutopic endometrium , 0.29 amol P03956 mRNA and 0.42 fmol P08253 mRNA per fmol P04406 mRNA were found . Relative P03956 mRNA concentrations increased strongly after culture on the P62158 , while P08253 mRNA levels were nearly unaltered . This differential regulation suggests different roles of these enzymes in the angiogenesis of ectopic endometrial fragments and during the development of endometriosis . 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 .