MH_dev_314

Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 SUB 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 SGLT 2 or P13866 ; ( 3 ) H - 2 - deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2 - electrode voltage clamp recording of oocytes expressing human SGLT 3 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 SUB dose-dependently decreased BG concentrations in db / db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA 1c ) 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 SUB 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 .

Proliferation of endothelial cell on polytetrafluoroethylene vascular graft materials carried P15692 gene plasmid . OBJECTIVE : To investigate whether vascular endothelial growth factor ( P15692 ) gene plasmid carried by polytetrafluoroethylene ( PTFE ) vascular graft materials could transfect endothelial cells ( ECs ) and promote their growth . METHODS : PTFE vascular graft materials carried with pCDI-hVEGF ( 121 ) , pCDI or pEGFP were incubated in DB03754 MEN - buffer solution and the values of optical density of 260 nm at different time were plotted , then the DNA controlled release curve was made . ECs derived from human umbilical vein were seeded on the pCDI-hVEGF ( 121 ) / pCDI / pEGFP-PTFE materials or tissue culture plates , ECs numbers were counted and P15692 protein concentrations at different time were measured by enzyme-linked immunoadsorbent assay method . Green fluorescent protein ( GFP ) expression in ECs on pEGFP-PTFE materials was examined with fluorescence microscopy . RESULTS : The controlled release curve showed that the gene released from PTFE materials was rapid within 8 h , then slowed down and that the gene released continuously even after 72 h . At 24 , 72 and 120 h , ECs number and proliferation rate of pCDI-hVEGF ( 121 ) - PTFE materials were higher than those of pCDI or pEGFP-PTFE materials ( P < 0.05 ) . P15692 protein concentration of pCDI-hVEGF ( 121 ) - PTFE materials was higher than that of pCDI or pEGFP-PTFE materials at 6 , 24 , 72 and 120 h ( P < 0.01 ) . GFP expression in ECs on the pEGFP-PTFE materials could be detected by fluorescence microscopy . CONCLUSION : PTFE graft can be used as a carrier of P15692 gene plasmid , P15692 gene carried by PTFE can transfect ECs and promote ECs growth .

Identification of immunodominant T cell epitopes of human glutamic acid decarboxylase 65 by using HLA-DR ( alpha 1 * 0101 , beta 1 * 0401 ) transgenic mice . DB00142 MEN decarboxylase isoform 2 ( Q05329 ; EC 4.1 . 1.15 ) has been identified as a key target autoantigen of insulin-dependent diabetes mellitus ( IDDM ) . IDDM is genetically associated with the major histocompatibility complex ( MHC ) , and particular alleles from the HLA-DQ and HLA-DR loci contribute to disease . Among DR4 subtypes , HLA - Q8IUH3 * 0401 , HLA - Q8IUH3 * 0402 , and HLA - Q8IUH3 * 0405 alleles lend susceptibility , while HLA - Q8IUH3 * 0403 confers protection . We have utilized HLA-DR ( alpha 1 * 0101 , beta 1 * 0401 ) ( hereafter referred to as DR0401 ) , human P01730 , murine class II null triple transgenic mice and recombinant Q05329 to generate T cell hybridomas , and we have used overlapping sets of peptides to map the immunodominant epitopes of this autoantigen . We have identified 10 immunogenic regions for Q05329 , of which 6 are recognized by multiple hybridomas . These epitopes are also generated by human antigen-presenting cells and their presentation is DR0401 restricted , as shown by the use of typed human lymphoblastoid cell lines and antibody blocking experiments . Immunodominant Q05329 epitopes defined in transgenic mice correspond to Q05329 regions previously shown to elicit T cell responses specifically in DR0401 IDDM patients , underscoring the validity of this approach . Interestingly , although the major epitopes contain DR0401 binding motifs , one of the epitopes contains a DR0405 motif .

Estradiol increases cell growth in human astrocytoma cell lines through ERα activation and its interaction with Q15788 and Q9Y6Q9 coactivators . Estradiol ( E2 ) regulates several cellular functions through the interaction with estrogen receptor subtypes , ERα and ERβ , which present different functional and regulation properties . ER subtypes have been identified in human astrocytomas , the most common and aggressive primary brain tumors . We studied the role of ER subtypes in cell growth of two human astrocytoma cell lines derived from tumors of different evolution grades : U373 and D54 ( grades III and IV , respectively ) . E2 significantly increased the number of cells in both lines and the co-administration with an ER antagonist ( ICI 182 , 780 ) significantly blocked E2 effects . ERα was the predominant subtype in both cell lines . E2 and ICI 182 , 780 down-regulated ERα expression . The number of U373 and D54 cells significantly increased after PPT ( ERα agonist ) treatment but not after DPN ( ERβ agonist ) one . To determine the role of Q15788 and Q9Y6Q9 coactivators in ERα induced cell growth , we silenced them with RNA interference . Coactivator silencing blocked the increase in cell number induced by PPT . The content of proteins involved in proliferation and metastasis was also determined after PPT treatment . Western blot analysis showed that in U373 cells the content of PR isoforms ( PR-A and PR-B ) , P00533 , P15692 and cyclin D1 increased after PPT treatment while in D54 cells only the content of P00533 was increased . Our results demonstrate that E2 induces cell growth of human astrocytoma cell lines through ERα and its interaction with Q15788 and Q9Y6Q9 and also suggest differential roles of ERα on cell growth depending on astrocytoma grade .

Tandutinib , an oral , small-molecule inhibitor of P36888 for the treatment of AML and other cancer indications . An improved understanding of acute myelogenous leukemia ( AML ) over the past two decades has led to a characterization of associated recurring cytogenetic abnormalities . AML is often driven by the overexpression or constitutive activation of receptor tyrosine kinases such as P36888 ( P36888 ) , which serves as a good therapeutic target . Millennium Pharmaceuticals Inc ' s DB05465 MEN ( DB05465 MEN ) is an orally active inhibitor of P36888 kinase and family members P09619 beta and c-Kit . Tandutinib inhibited P36888 phosphorylation , downstream signaling and malignant growth in vitro and in animal models . The drug exhibited limited activity as a single agent in phase I and II clinical trials in patients with AML and myelodysplastic syndrome , but displayed promising antileukemic activity ( 90 % complete remissions ) in a phase I / II trial in patients with newly diagnosed AML when administered in combination with cytarabine and daunorubicin . Phase II clinical trials for DB05465 MEN are ongoing in patients with AML or renal cell carcinoma .

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 ) .

Steroid receptor coactivator 2 is required for female fertility and mammary morphogenesis : insights from the mouse , relevance to the human . Although the importance of the progesterone receptor ( PR ) to female reproductive and mammary gland biology is firmly established , the coregulators selectively co-opted by PR in these systems have not been clearly delineated . A selective gene-knockout approach applied to the mouse , which abrogates gene function only in cell types that express PR , recently disclosed steroid receptor coactivator 2 ( P12931 - 2 , also known as Q06418 - 2 or Q9Y3R0 ) to be an indispensable coregulator for uterine and mammary gland responses that require progesterone . Uterine cells positive for PR ( but devoid of P12931 - 2 ) were found to be incapable of facilitating embryo implantation , a necessary first step toward the establishment of the materno-fetal interface . Importantly , such an implantation defect is not exhibited by knockouts for Q15788 or Q9Y6Q9 , underscoring the unique coregulator importance of P12931 - 2 in peri-implantation biology . Moreover , despite normal levels of PR , Q15788 and Q9Y6Q9 , progesterone-dependent branching morphogenesis and alveologenesis fails to occur in the murine mammary gland in the absence of P12931 - 2 , thereby establishing a critical coregulator role for P12931 - 2 in signaling cascades that mediate progesterone-induced mammary epithelial proliferation . Finally , the recent detection of P12931 - 2 in the human endometrium and breast suggests that this coregulator may represent a new clinical target for the future management of female reproductive health and / or breast cancer .

DB00115 MEN is a strong determinant of low methionine synthase activity and DNA hypomethylation in gastrectomized rats . BACKGROUND / AIMS : The respective influence of folate and vitamin B12 deficiency on Q99707 activity and transcription , and on DNA methylation is not clearly established . The aim of this study was to assess the respective influence of folate and vitamin B12 deficiency on Q99707 transcription and activity , and on DNA methylation . METHODS : Sixty-one rats were administered normal diet or diet deficient in choline , methionine , folic acid and vitamin B12 . Forty-seven of them underwent total gastrectomy or ileal resection . RESULTS : Low vitamin B12 was observed only in gastrectomized rats . Low folate was observed in rats under deficient diet . Total Q99707 activity ( holo - + apoenzyme ) was lowered only with vitamin B12 level < 200 pmol / l ( p= 0.0002 ) , while the ratios of total vs . holo - Q99707 activity and of transcripts Q99707 vs . P04406 ( RT-PCR ) were unchanged . DB00115 MEN was the single determinant of low Q99707 ( lower quartile , odds ratio = 15.75 , p= 0.0017 ) . Low Q99707 and low vitamin B12 were the two determinants of DNA hypomethylation ( lower quartile ) ( odds ratio = 17.07 , p= 0.0006 , and odds ratio = 7.31 , p= 0.006 , respectively ) . CONCLUSION : DB00115 MEN affects Q99707 expression by a non-transcriptional mechanism different from a protective effect on Q99707 proteolysis . It is also a strong determinant of DNA hypomethylation .

The transcriptional coactivator DRIP / mediator complex is involved in vitamin D receptor function and regulates keratinocyte proliferation and differentiation . Mediator is a multisubunit coactivator complex that facilitates transcription of nuclear receptors . We investigated the role of the mediator complex as a coactivator for vitamin D receptor ( P11473 ) in keratinocytes . Using P11473 affinity beads , the vitamin D receptor interacting protein ( DRIP ) / mediator complex was purified from primary keratinocytes , and its subunit composition was determined by mass spectrometry . The complex included core subunits , such as Q15648 / MED 1 ( MED 1 ) , that directly binds to P11473 . Additional subunits were identified that are components of the RNA polymerase II complex . The functions of different mediator components were investigated by silencing its subunits . The core subunit MED 1 facilitates P11473 activity and regulating keratinocyte proliferation and differentiation . A newly described subunit Q13503 also has a role in promoting keratinocyte proliferation and differentiation , whereas Q9BTT4 has an inhibitory role . Blocking MED 1 / Q13503 expression caused hyperproliferation of keratinocytes , accompanied by increases in mRNA expression of the cell cycle regulator cyclin D1 and / or glioma-associated oncogene homolog . Blocking MED 1 or Q13503 expression also resulted in defects in calcium-induced keratinocyte differentiation , as indicated by decreased expression of differentiation markers and decreased translocation of P12830 to the membrane . These results show that keratinocytes use the transcriptional coactivator mediator to regulate P11473 functions and control keratinocyte proliferation and differentiation .

Role of antispasmodics in the treatment of irritable bowel syndrome . Irritable bowel syndrome ( IBS ) is a long-lasting , relapsing disorder characterized by abdominal pain / discomfort and altered bowel habits . Intestinal motility impairment and visceral hypersensitivity are the key factors among its multifactorial pathogenesis , both of which require effective treatment . Voltage-gated calcium channels mediate smooth muscle contraction and endocrine secretion and play important roles in neuronal transmission . Antispasmodics are a group of drugs that have been used in the treatment of IBS for decades . DB01616 MEN citrate , a spasmolytic , decreases the sensitivity of smooth muscle contractile proteins to calcium , and it is a selective P08908 receptor antagonist . DB01616 MEN , in combination with simethicone , has been demonstrated to effectively reduce abdominal pain and discomfort in a large placebo-controlled trial . Mebeverine is a musculotropic agent that potently blocks intestinal peristalsis . Non-placebo-controlled trials have shown positive effects of mebeverine in IBS regarding symptom control ; nevertheless , in recent placebo-controlled studies , mebeverine did not exhibit superiority over placebo . Otilonium bromide is poorly absorbed from the GI tract , where it acts locally as an L-type calcium channel blocker , an antimuscarinic and a tachykinin NK2 receptor antagonist . Otilonium has effectively reduced pain and improved defecation alterations in placebo-controlled trials in IBS patients . DB09090 bromide is also an L-type calcium channel blocker that acts locally in the GI tract . DB09090 improves motility disorders and consequently reduces stool problems in IBS patients . Phloroglucinol and trimethylphloroglucinol are non-specific antispasmodics that reduced pain in IBS patients in a placebo-controlled trial . Antispasmodics have excellent safety profiles . T-type calcium channel blockers can abolish visceral hypersensitivity in animal models , which makes them potential candidates for the development of novel therapeutic agents in the treatment of IBS .

2 ( 3H ) - benzoxazolone and bioisosters as " privileged scaffold " in the design of pharmacological probes . The 2 ( 3H ) - benzoxazolone heterocycle and its bioisosteric surrogates ( such as 2 ( 3H ) - benzothiazolinone , benzoxazinone , etc . ) have received considerable attention from the medicinal chemists owing to their capacity to mimic a phenol or a catechol moiety in a metabolically stable template . These heterocycles and pyrocatechol have indeed similar pKa ' s , electronic charge distribution , and chemical reactivity . Therapeutic applications of this template are very broad , and range from analgesic anti-inflammatory compounds ( including P37231 antagonists ) to antipsychotic and neuroprotective anticonvulsant compounds . High affinity ligands have been obtained also for dopaminergic ( D2 and D4 ) , serotoninergic ( P08908 and P28223 ) , sigma - 1 and sigma - 2 receptors . Owing to the high number of positive hits encountered with this heterocycle and its congeners , 2 ( 3H ) - benzoxazolone template certainly deserves the title of " privileged scaffold " in medicinal chemistry .

P04150 interacting protein - 1 restores glucocorticoid responsiveness in steroid-resistant airway structural cells . Glucocorticoid ( GC ) insensitivity represents a profound challenge in managing patients with asthma . The mutual inhibition of transcriptional activity between GC receptor ( GR ) and other regulators is one of the mechanisms contributing to GC resistance in asthma . We recently reported that interferon regulatory factor ( Q969Q1 ) - 1 is a novel transcription factor that promotes GC insensitivity in human airway smooth muscle ( P17405 ) cells by interfering with GR signaling ( Tliba et al . , Am J Respir Cell Mol Biol 2008 ; 38:463- 472 ) . Here , we sought to determine whether the inhibition of GR function by P10914 involves its interaction with the transcriptional co-regulator GR-interacting protein 1 ( Q9Y3R0 ) , a known GR transcriptional co-activator . We here found that siRNA-mediated Q9Y3R0 depletion attenuated P10914 - dependent transcription of the luciferase reporter construct and the mRNA expression of an P10914 - dependent gene , P28907 . In parallel experiments , Q9Y3R0 silencing significantly reduced GR-mediated transactivation activities . Co-immunoprecipitation and Q86UG4 pull-down assays showed that Q9Y3R0 , through its repression domain , physically interacts with P10914 identifying Q9Y3R0 as a bona fide transcriptional co-activator for P10914 . Interestingly , the previously reported inhibition of GR-mediated transactivation activities by either P01375 and P01579 treatment or P10914 overexpression was fully reversed by increasing cellular levels of Q9Y3R0 . Together , these data suggest that the cellular accumulation of P10914 may represent a potential molecular mechanism mediating altered cellular response to GC through the depletion of Q9Y3R0 from the GR transcriptional regulatory complexes .

Potential utility of telmisartan , an angiotensin II type 1 receptor blocker with peroxisome proliferator-activated receptor-gamma ( P37231 ) - modulating activity for the treatment of cardiometabolic disorders . The metabolic syndrome is strongly associated with insulin resistance and consists of a constellation of factors such as hypertension and hyperlipidemia that raise the risk for cardiovascular diseases and diabetes mellitus . There is widespread agreement that the renin-angiotensin system ( DB01367 ) plays a pivotal role in the pathogenesis of insulin resistance and cardiovascular disease in diabetes . Indeed , large clinical trials have demonstrated substantial benefit of the blockade of this system for cardiovascular end-organ protection . Thus the blockade of the DB01367 may be a promising strategy for the treatment of the patients with the metabolic syndrome . Although several types of angiotensin II type 1 ( AT ( 1 ) ) receptor blockers ( ARBs ) are commercially available for the treatment of patients with hypertension , we have recently found that telmisartan ( DB00966 MENMAX DB00966 MEN ) could have the strongest binding affinity to AT ( 1 ) receptor . Further , telmisartan is reported to act as a partial agonist of peroxisome proliferator-activated receptor-gamma ( P37231 ) . These observations suggest that , due to its unique P37231 - modulating activity , telmisartan may be one of the most promising sartans for the treatment of cardiometabolic disorders . In this paper , we reviewed the potential utility of telmisartan in insulin resistance and vascular complications in diabetes .

A guide to picking the most selective kinase inhibitor tool compounds for pharmacological validation of drug targets . To establish the druggability of a target , genetic validation needs to be supplemented with pharmacological validation . Pharmacological studies , especially in the kinase field , are hampered by the fact that many reference inhibitors are not fully selective for one target . Fortunately , the initial trickle of selective inhibitors released in the public domain has steadily swelled into a stream . However , rationally picking the most selective tool compound out of the increasing amounts of available inhibitors has become progressively difficult due to the lack of accurate quantitative descriptors of drug selectivity . A recently published approach , termed ' selectivity entropy ' , is an improved way of expressing selectivity as a single-value parameter and enables rank ordering of inhibitors . We provide a guide to select the best tool compounds for pharmacological validation experiments of candidate drug targets using selectivity entropy . In addition , we recommend which inhibitors to use for studying the biology of the 20 most investigated kinases that are clinically relevant : Abl ( P00519 ) , P31749 , Q9UM73 , Aurora A / B , CDKs , MET , P07333 ( P07333 ) , P00533 , P36888 , P04626 ( P04626 ) , O14920 ( O14920 ) , O60674 / 3 , P45983 / 2/3 ( P45983 / 9/10 ) , Q02750 / 2 , P53350 , PI3Ks , p38α ( Q16539 ) , P15056 , P12931 and P35968 ( P35968 ) .

Mislocalization of cell-cell adhesion complexes in tamoxifen-resistant breast cancer cells with elevated c-Src tyrosine kinase activity . c-Src activation has been implicated in metastasis of tamoxifen-resistant breast cancer . Here we investigated how c-Src activity affects cell adhesion using a tamoxifen-resistant variant of MCF - 7 cells ( Q99707 - 3 ) containing elevated c-Src activity . In Q99707 - 3 cells , adhesion proteins beta-catenin and P12830 are mislocalized , forming novel structures perpendicular to cell-cell junctions . c-Src is associated with beta-catenin / P12830 complexes and beta-catenin tyrosine phosphorylation is enhanced . Blocking c-Src tyrosine kinase activity decreased beta-catenin tyrosine phosphorylation and restored localization of beta-catenin and P12830 at cell-cell junctions . These findings suggest that inhibition of c-Src signaling may prevent metastasis of tamoxifen-resistant breast cancer .

DB05767 MEN ( Andrographis paniculata extract ) prevents development of murine colitis by inhibiting T-cell proliferation and Q8IXH7 / TH17 responses . BACKGROUND : Extracts of the plant Andrographis paniculata have been used to treat inflammatory diseases in Asian countries . A recent double-blind , placebo-controlled trial of DB05767 MEN ( A . paniculata extract ) has demonstrated its safety and effectiveness for induction of clinical response , remission , and mucosal healing in patients with mild to moderate ulcerative colitis ( UC ) . We aimed to determine if DB05767 MEN could prevent the development of T-cell-dependent murine colitis and to define its in vivo mechanism ( s ) of action . METHODS : CD ( + ) 4CD45RB ( high ) T cells were transferred into Rag 1 ( - / - ) mice and gavaged daily with DB05767 MEN or methyl cellulose ( MC ) . Severity of colitis was evaluated by weight loss , histology , and cytokine expression . RESULTS : Mice treated with MC developed colitis within 4-7 weeks , as evaluated by weight loss , and severe intestinal inflammation . DB05767 MEN - treated mice did not lose weight and displayed only very mild intestinal inflammation . P01375 alpha ( P01375 - α ) , interleukin ( IL ) - 1β , interferon-gamma ( IFN-γ ) , and Q9GZX6 expression were significantly decreased in DB05767 MEN - treated mice . We observed higher percentages of naïve P01730 ( + ) T cells in the lamina propria of DB05767 MEN - treated mice . At early timepoints DB05767 MEN - treated mice have significantly reduced splenic cell counts , reduced P01730 ( + ) , and Q16552 ( + ) , and IFN-γ T ( + ) cells . Furthermore , DB05767 MEN inhibited the proliferation of P01730 T cells and differentiation into Q8IXH7 / TH17 cells in vitro . CONCLUSIONS : DB05767 MEN inhibits the development of chronic colitis by affecting early T-cell proliferation , differentiation , and TH ( 1 ) / TH ( 17 ) responses in a T-cell-driven model of colitis , presenting a unique mechanism of action . Our data suggest that DB05767 MEN could be an attractive herbal therapeutic for inflammatory bowel disease .

Phosphatase P35813 negatively regulates P-TEFb function in resting P01730 ( + ) T cells and inhibits HIV - 1 gene expression . BACKGROUND : Processive elongation of the integrated HIV - 1 provirus is dependent on recruitment of P-TEFb by the viral Tat protein to the viral TAR RNA element . P-TEFb kinase activity requires phosphorylation of Thr 186 in the T-loop of the P50750 subunit . In resting P01730 + T cells , low levels of T-loop phosphorylated P50750 are found , which increase significantly upon activation . This suggests that the phosphorylation status of the T-loop is actively regulated through the concerted actions of cellular proteins such as DB00133 / DB00156 phosphatases . We investigated the role of phosphatase P35813 in regulating P50750 T-loop phosphorylation and its effect on HIV - 1 proviral transcription . RESULTS : We found that overexpression of P35813 inhibits HIV - 1 gene expression during viral infection and this required P35813 catalytic function . Using an artificial CDK tethering system , we further found that P35813 inhibits P50750 , but not P49336 mediated activation of the HIV - 1 LTR . SiRNA depletion of P35813 in resting P01730 + T cells increased the level of P50750 T-loop phosphorylation and enhanced HIV - 1 gene expression . We also observed that P35813 protein levels are relatively high in resting P01730 + T cells and are not up-regulated upon T cell activation . CONCLUSIONS : Our results establish a functional link between HIV - 1 replication and modulation of P50750 T-loop phosphorylation by P35813 . P35813 represses HIV - 1 gene expression by inhibiting P50750 T-loop phosphorylation , thus reducing the amount of active P-TEFb available for recruitment to the viral LTR . We also infer that P35813 enzymatic activity in resting and activated P01730 + T cells are likely regulated by as yet undefined factors .

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-β 1 , TGF-β 2 , 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-β 1 , TGF-β 2 , 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 .

Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 MEN , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 MEN [ ( R ) - ( - ) - 2 - ( 4 - bromo - 2 - fluorobenzyl ) -1,2 , 3,4- tetrahydropyrrolo [ 1,2- a ] pyrazine - 4 - spiro - 3 ' - pyrrolidine -1,2 ' , 3,5 ' - tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K ( i ) = 10 ( - ) ( 10 ) M ) 2000 - fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R - and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP ( + ) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP ( + ) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP ( + ) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and / or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4 - bromo - 2 - fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) .

A set of consensus mammalian mediator subunits identified by multidimensional protein identification technology . The Mediator is a multiprotein transcriptional coactivator that is expressed ubiquitously in eukaryotes from yeast to mammals and is required for induction of RNA polymerase II ( pol II ) transcription by DNA binding transcription factors . In the work described here , we exploit multidimensional protein identification technology ( MudPIT ) to carry out a proteomic analysis of the subunit composition of the mammalian Mediator complex . By comparing MudPIT data sets obtained from six independent Mediator preparations immunoaffinity purified through their Nut 2 ( Q9BTT4 ) , Med 25 ( Q9NWA0 ) , Intersex ( Q9NX70 ) , A0JLT2 ( A0JLT2 ) , AK007855 ( Q9H204 ) , or CRSP 70 ( O95402 ) subunits , we identify a set of consensus mammalian Mediator subunits . In addition , we identify as Mediator-associated proteins the P49336 - like cyclin-dependent kinase CDK 11 and the Q9UHV7 - like Q71F56 protein ( Q71F56 ) , which is mutated in patients with the congenital heart defect transposition of the great arteries ( TGA ) .

Estrogen receptors mediate rapid activation of phospholipase C pathway in the rat endometrium . The aim of the present study was to investigate the activation of rapid signaling events by 17β - estradiol in the rat uterus . 17β - Estradiol induced a rapid increase of total [ 3H ] - inositol phosphate accumulation in the whole uterus and endometrium , but not in the myometrium . The effect of 17β - estradiol in the endometrium was blocked by phospholipase C ( P98160 ) inhibitor ( U73122 ) , estrogen receptors antagonist ( DB00947 MEN ) , exportin O14980 inhibitor ( leptomycin B ) and selective inhibitor of the P12931 family of protein tyrosine kinases ( Q99463 ) . Furthermore , a selective agonist of P03372 ( PPT ) and a selective agonist of Q99527 ( G - 1 ) also induced a rapid increase of total [ ( 3 ) H ] - inositol phosphate accumulation in the endometrium . The G - 1 effects were blocked by Q99527 antagonist ( G - 15 ) . 17β - Estradiol and G - 1 promoted an additive effect on total [ 3H ] - inositol phosphate accumulation . In conclusion , the present results indicate that a rapid activation of the P98160 - mediated phosphoinositide hydrolysis occurred in the rat endometrium after 17β - estradiol stimulation , and this effect was mediated by P03372 that underwent nuclear export after hormone stimulation , and that Q99527 activation may play an additive role for this response . These rapid actions might be one of the key steps that mediate the estrogen-dependent activation of cellular events in the endometrium .

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 .