interacts_with DB00700?
Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Cbl-b is a negative regulator of inflammatory cytokines produced by IgE-activated mast cells . c-Cbl and Cbl-b E3 ubiquitin ligases are abundantly expressed in hemopoietic cells where they negatively regulate the activity and levels of many cell surface receptors and associated signaling molecules . By comparing bone marrow-derived mast cells from c-Cbl and Cbl-b-deficient mice it has recently been shown that Cbl-b is the dominant family member for negatively regulating signaling responses from high-affinity IgE receptors . In this study , we suggest that a possible reason for the greater enhancement of IgE receptor signaling in Cbl-b-deficient mice is the relatively higher levels of Cbl-b protein over c-Cbl in mast cells compared with other hemopoietic cells . We also directly compare mast cells from c-Cbl and Cbl-b-deficient mice and find that loss of Cbl-b , but not c-Cbl , increases cell growth , retards receptor internalization , and causes the sustained tyrosine phosphorylation of Syk and its substrates . However , loss of Cbl-b does not enhance the activation of P29323 or Akt , nor does it promote a greater calcium response . Furthermore , loss of Cbl-b or c-Cbl does not increase levels of the Syk or Lyn protein tyrosine kinases . Most notable , however , is the extremely large increase in the production of proinflammatory cytokines P01375 , P05231 , and P13500 by Cbl-b(-/-) mast cells compared with levels produced by c-Cbl(-/-) or wild-type cells . This marked induction , which appears to be restricted to these three cytokines , is dependent on IgE receptor activation and correlates with enhanced O15111 phosphorylation . Thus , Cbl-b functions as a potent negative regulator of cytokines that promote allergic and inflammatory reactions . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Role of cardiovascular aldosterone in hypertension . DB04630 plays an important role in the pathogenesis of cardiovascular disease . We have reported that aldosterone is synthesized in cardiovascular tissues and local aldosterone synthesis plays important roles for hypertension and cardiac hypertrophy . High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP . Plasma aldosterone concentrations and P06703 were decreased by high salt intake in SHRSP . DB04630 production , the expression of P19099 mRNA and angiotensin II receptor AT1R mRNA in blood vessels were significantly increased by high salt intake . These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the vascular tissue in SHRSP , which may contribute to the development of malignant hypertension in salt-loaded SHRSP . However , there are several reports of conflicting data . P08235 ( MR ) binding is tightly regulated by the enzyme 11beta-hydroxysteroid dehydrogenase type 2 ( 11beta-HSD2 ) which selectively metabolizes glucocorticoids to inactive metabolites , thus allowing for MR activation by aldosterone . We have reported that decreased 11beta-HSD2 in blood vessels in Dahl salt-sensitive ( DS ) rats , a model for salt-sensitive hypertension . Local aldosterone excess may play a significant role in the salt sensitivity and development of hypertension . High sodium intake decreased circulating rennin-angiotensin-aldosterone system and increased blood pressure and cardiac hypertrophy in DS rats , which were prevented by the treatment with a selective MR antagonist , eplerenone . DB00700 also improved endothelial nitric oxide synthase ( P29474 ) activity and P29474 mRNA expression in blood vessels in DS rats . These results further suggest that not only circulating aldosterone but also local aldosterone is of critical importance in the pathophysiology of cardiovascular disorders . The expression of the solute carriers Q14973 and O75051 -1 is regulated by cholesterol in HepG2 cells . Drug disposition and response are greatly determined by the activities of drug-metabolizing enzymes and transporters . While the knowledge in terms of CYP enzymes and efflux ABC transporters ( such as P08183 , P-glycoprotein ) is quite extensive , influx transporters are increasingly being unveiled as key contributors to the process of drug disposition . There is little information on the regulation of these proteins in human cells , especially as regards the effect of endogenous compounds . In this study , we analysed the expression of P08684 and three uptake transporters Q14973 ( Q14973 ) , P46721 / P46721 ( P46721 ) and O75051 -1 ( O15245 ) in HepG2 cells following treatment with cholesterol . While P08684 and P46721 expression was unaffected , cholesterol treatment led to increased levels of Q14973 and O75051 -1 mRNAs . Alterations in the functional characteristics and/or expression levels of drug transporters in the liver may conceivably contribute to the variability in drug oral bioavailability often observed in the clinical settings . Periadventitial adipose tissue impairs coronary endothelial function via P05771 -dependent phosphorylation of nitric oxide synthase . Endogenous periadventitial adipose-derived factors have been shown to contribute to coronary vascular regulation by impairing endothelial function through a direct inhibition of endothelial nitric oxide synthase ( P29474 ) . However , our understanding of the underlying mechanisms remains uncertain . Accordingly , this study was designed to test the hypothesis that periadventitial adipose tissue releases agents that attenuate coronary endothelial nitric oxide production via a protein kinase C ( PKC ) -beta-dependent mechanism . Isometric tension studies were conducted on isolated canine circumflex coronary arteries with and without natural amounts of periadventitial adipose tissue . Adipose tissue significantly diminished coronary endothelial-dependent vasodilation and nitric oxide production in response to bradykinin and acetylcholine . The selective inhibition of endothelial P05771 with ruboxistaurin ( 1 microM ) abolished the adipose-induced impairment of bradykinin-mediated coronary vasodilation and the endothelial production of nitric oxide . Western blot analysis revealed a significant increase in P29474 phosphorylation at the inhibitory residue DB00156 (495) in arteries exposed to periadventitial adipose tissue . This site-specific phosphorylation of P29474 was prevented by the inhibition of P05771 . These data demonstrate that periadventitial adipose-derived factors impair coronary endothelial nitric oxide production via a P05771 -dependent , site-specific phosphorylation of P29474 at DB00156 (495) . Effects of dutasteride on the expression of genes related to androgen metabolism and related pathway in human prostate cancer cell lines . Androgens play an important role in controlling the growth of the normal prostate gland and in the pathogenesis of benign prostate hyperplasia , and prostate cancer . Although testosterone is the main androgen secreted from the testes , dihydrotestosterone ( DB02901 ) , a more potent androgen converted from testosterone by 5alpha-reductase isozymes , type I and II , is the major androgen in the prostate cells . The aim of this study is to investigate the cellular and molecular effects of dutasteride , a potent inhibitor of 5alpha-reductase type I and type II , in androgen-responsive ( LNCaP ) and androgen-unresponsive ( DU145 ) human prostate cancer(PCa) cell lines . The expression pattern of 190 genes , selected on the basis of their proved or potential role in prostate cancerogenesis related to androgen signalling , were analysed using a low density home-made oligoarray ( AndroChip 2 ) . Our results show that dutasteride reduces cell viability and cell proliferation in both cell lines tested . AndroChip 2 gene signature identified in LNCaP a total of 11 genes differentially expressed ( FC > or= +/-1.5 ) . Eight of these genes , were overexpressed and three were underexpressed . Overexpressed genes included genes encoding for proteins involved in biosynthesis and metabolism of androgen ( P14061 ; P37058 ; P19099 ) , androgen receptor and androgen receptor co-regulators ( AR; P24385 ) , and signal transduction ( P04626 ; V- P62158 ; Q07889 ) whereas , underexpressed genes ( KLK3 ; P20151 ; Q15392 ) were androgen-regulated genes ( ARGs ) . No differentially expressed genes were scored in DU145 . Microarray data were confirmed by quantitative real-time PCR assay ( QRT-PCR ) . These data offer a selective genomic signature for dutasteride treatment in prostate epithelial cells and provide important insights in prostate cancer pathophysiology . T cell-mediated vascular dysfunction of human allografts results from P01579 dysregulation of NO synthase . Allograft vascular dysfunction predisposes to arteriosclerosis and graft loss . We examined how dysfunction develops in transplanted human arteries in response to circulating allogeneic T cells in vivo using immunodeficient murine hosts . Within 7-9 days , transplanted arteries developed endothelial cell ( EC ) dysfunction but remained sensitive to exogenous NO . By 2 weeks , the grafts developed impaired contractility and desensitization to NO , both signs of VSMC dysfunction . These T cell-dependent changes correlated with loss of P29474 and expression of P35228 -- the latter predominantly within infiltrating T cells . Neutralizing P01579 completely prevented both vascular dysfunction and changes in NOS expression ; neutralizing P01375 reduced P01579 production and partially prevented dysfunction . Inhibiting P35228 partially preserved responses to NO at 2 weeks and reduced graft intimal expansion after 4 weeks in vivo . In vitro , memory P01730 + T cells acted on allogeneic cultured ECs to reduce P29474 activity and expression of protein and mRNA . These effects required T cell activation by class II MHC antigens and costimulators ( principally lymphocyte function-associated antigen-3 , or LFA-3 ) on the ECs and were mediated by production of soluble mediators including P01579 and P01375 . We conclude that P01579 is a central mediator of vascular dysfunction and , through dysregulation of NOS expression , links early dysfunction with late arteriosclerosis . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . Some studies on spontaneous Hymenolepis diminuta infection in laboratory rats . Hymenolepis diminuta is a tapeworm that occurs worldwide . It is known to be found commonly in areas where large amounts of food grains or other dry feed products , which are the favorite foods for rats . Transmission of disease to human is uncommon ; however , it may be a serious threat for population who are living in rural areas which are suffering from excessive rodents . Here , this study had done on spontaneous H. diminuta infection in laboratory rats as a model . Out of thirty five adult laboratory rats investigated for parasitic diseases only nine ( 25.71 % ) were diagnosed positive for spontaneous H. diminuta infection . Four of them ( 44.44 % ) were found losing of weight and lacking of motility , while the others were normal . On microscopic examination , H. diminuta eggs had been found in their stool . On autopsy , small intestines were found to contain from 5-6 multi-segmented tapeworms in each rat . Histopathologically , intestinal lumen showed varying sections of H. diminuta segments with serrated borders . H. diminuta infection caused multiple mucosal ulcers with absence of intestinal villi from the surface epithelium and excessive mucin . Moreover , inflammatory cells infiltration in the connective tissue core of the villi . Furthermore , the Toluidine blue stain showed that there are Mastiocytosis . Additionally , there were goblet cells hyperplasia on using DB00233 . Moreover , there were high expression of cyclooxygenase 2 ( P35354 ) , tumor necrosis factor-α ( P01375 -α ) and inducible Nitric-Oxide Synthase ( iNOs ) . This implicate , strong correlation between P35354 , P01375 -α and iNOs expression and inflammation induced by H. diminuta . Effect of DB02527 on TGFbeta1-induced corneal keratocyte-myofibroblast transformation . PURPOSE : TGFbeta is the major mediator to induce myofibroblast differentiation in the corneal wound-healing process . Elevated DB02527 can reduce TGFbeta-induced fibrosis in other tissues . This study was conducted to determine whether elevated DB02527 can inhibit TGFbeta1-induced rabbit corneal keratocyte-myofibroblast transformation . METHODS : Primary isolated rabbit corneal keratocytes were cultured in serum-free medium . The effects of the adenylate cyclase agonist forskolin ( FSK ; 2 microM ) on TGFbeta1 ( 5 ng/mL ) -induced alpha-smooth muscle actin ( alpha-SMA ) expression was examined by immunofluorescence , flow cytometry , and immunochemistry 72 hours after treatment . The effects of TGFbeta+FSK on activated pSmad3 , CREB binding protein ( CBP ) , MAPKs , and RhoA were determined by coimmunoprecipitation and Western blot . RESULTS : FSK significantly reduced the myofibroblast phenotype and alpha-SMA expression induced by TGFbeta1 in rabbit corneal keratocytes . TGFbeta1 increased the phosphorylation of P29323 and P84022 . TGFbeta1-induced alpha-SMA expression was reduced by MEK inhibition ( U0126 ) ; however , the levels of pERK , pSmad3 , or the extent of the interaction between pSmad3 and CBP induced by TGFbeta1 were not affected by FSK . TGFbeta1 also activated RhoA and ROCK ( Y27632 ) inhibition reduced alpha-SMA expression . Activation of RhoA was significantly reduced by FSK . CONCLUSIONS : Raising DB02527 by FSK treatment inhibits the TGFbeta1-induced corneal myofibroblast transformation and alpha-SMA expression and thereby provides a promising method to control corneal fibrosis . The data suggest that DB02527 -dependent inhibition does not occur by altering Smads or MAPK signaling , but possibly by reducing the activation of RhoA . Dacomitinib ( PF-00299804 ) , an irreversible Pan-HER inhibitor , inhibits proliferation of P04626 -amplified breast cancer cell lines resistant to trastuzumab and lapatinib . The human P01133 ( HER ) family of receptors has been pursued as therapeutic targets in breast cancer and other malignancies . DB00072 and lapatinib are standard treatments for P04626 -amplified breast cancer , but a significant number of patients do not respond or develop resistance to these drugs . Here we evaluate the in vitro activity of dacomitinib ( PF-00299804 ) , an irreversible small molecule pan-HER inhibitor , in a large panel of human breast cancer cell lines with variable expression of the HER family receptors and ligands , and with variable sensitivity to trastuzumab and lapatinib . Forty-seven human breast cancer and immortalized breast epithelial lines representing the known molecular subgroups of breast cancer were treated with dacomitinib to determine IC(50) values . P04626 -amplified lines were far more likely to respond to dacomitinib than nonamplified lines ( RR , 3.39 ; P < 0.0001 ) . Furthermore , P04626 mRNA and protein expression were quantitatively associated with response . Dacomitinib reduced the phosphorylation of P04626 , P00533 , Q15303 , AKT , and P29323 in the majority of sensitive lines . Dacomitinib exerted its antiproliferative effect through a combined G(0)-G(1) arrest and an induction of apoptosis . Dacomitinib inhibited growth in several P04626 -amplified lines with de novo and acquired resistance to trastuzumab . Dacomitinib maintained a high activity in lines with acquired resistance to lapatinib . This study identifies P04626 -amplified breast cancer lines as most sensitive to the antiproliferative effect of dacomitinib and provides a strong rationale for its clinical testing in P04626 -amplified breast cancers resistant to trastuzumab and lapatinib . 20-hydroxy-5,8,11,14-eicosatetraenoic acid mediates endothelial dysfunction via O15111 -dependent endothelial nitric-oxide synthase uncoupling . Endothelial dysfunction and activation occur in the vasculature and are believed to contribute to the pathogenesis of cardiovascular diseases . We have shown that 20-hydroxy-5,8,11,14-eicosatetraenoic acid ( 20-HETE ) , a cytochrome P450 4A-derived eicosanoid that promotes vasoconstriction in the microcirculation , uncouples endothelial nitric-oxide synthase ( P29474 ) and reduces nitric oxide ( NO ) levels via the dissociation of the 90-kDa heat shock protein ( HSP90 ) from P29474 . It also causes endothelial activation by stimulating nuclear factor-kappaB ( NF-kappaB ) and increasing levels of pro-inflammatory cytokines . In this study , we examined signaling mechanisms that may link 20-HETE-induced endothelial dysfunction and activation . Under conditions in which 20-HETE inhibited NO production , it also stimulated inhibitor of NF-kappaB ( IkappaB ) phosphorylation . Both effects were prevented by inhibition of tyrosine kinases and mitogen-activated protein kinase ( MAPK ) /extracellular signal-regulated kinase ( P29323 ) . It is noteworthy that inhibitor of O15111 ( IKK ) activity negated the 20-HETE-mediated inhibition of NO production . Immunoprecipitation experiments revealed that treatment of ionophore-stimulated cells with 20-HETE brings about a decrease in HSP90- P29474 association and an increase in HSP90-IKKbeta association , suggesting that the activation by 20-HETE of NF-kappaB is linked to its action on P29474 . Furthermore , addition of inhibitors of tyrosine kinase MAPK and IKK restored the 20-HETE-mediated impairment of acetylcholine-induced relaxation in rat renal interlobar arteries . The results indicate that 20-HETE mediates P29474 uncoupling and endothelial dysfunction via the activation of tyrosine kinase , MAPK , and IKK , and these effects are linked to 20-HETE-mediated endothelial activation . In vivo magnetomotive optical molecular imaging using targeted magnetic nanoprobes . Dynamic magnetomotion of magnetic nanoparticles ( MNPs ) detected with magnetomotive optical coherence tomography ( MM- O75051 ) represents a new methodology for contrast enhancement and therapeutic interventions in molecular imaging . In this study , we demonstrate in vivo imaging of dynamic functionalized iron oxide MNPs using MM- O75051 in a preclinical mammary tumor model . Using targeted MNPs , in vivo MM- O75051 images exhibit strong magnetomotive signals in mammary tumor , and no significant signals were measured from tumors of rats injected with nontargeted MNPs or saline . The results of in vivo MM- O75051 are validated by Q9BWK5 , ex vivo MM- O75051 , DB06783 staining of histological sections , and immunohistochemical analysis of excised tumors and internal organs . The MNPs are antibody functionalized to target the human epidermal growth factor receptor 2 ( P04626 neu ) protein . Fc-directed conjugation of the antibody to the MNPs aids in reducing uptake by macrophages in the reticulo-endothelial system , thereby increasing the circulation time in the blood . These engineered magnetic nanoprobes have multifunctional capabilities enabling them to be used as dynamic contrast agents in MM- O75051 and Q9BWK5 . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Concise prediction models of anticancer efficacy of 8 drugs using expression data from 12 selected genes . We developed concise , accurate prediction models of the in vitro activity for 8 anticancer drugs ( DB00544 , DB00515 , DB00305 , DOX , CPT-11 , SN-38 , TXL and TXT ) , along with individual clinical responses to DB00544 using expression data of 12 genes . We first performed cDNA microarray analysis and MTT assay of 19 human cancer cell lines to sort out genes which were correlative in expression levels with cytotoxicities of the 8 drugs ; we selected 13 genes with proven functional significance to drug sensitivity from a huge number of potent prediction marker genes . The correlation significance of each was confirmed using expression data quantified by real-time RT-PCR , and finally 12 genes ( P08183 , Q9UNQ0 , P10632 , P08684 , Q12882 , P09211 , P16455 , P15559 , P16435 , P11388 , P07437 and P04818 ) were selected as more reliable predictors of drug response . Using multiple regression analysis , we fixed 8 prediction formulae which embraced the variable expressions of the 12 genes and arranged them in order , to predict the efficacy of the drugs by referring to the value of Akaike 's information criterion for each sample . These formulae appeared to accurately predict the in vitro efficacy of the drugs . For the first clinical application model , we fixed prediction formulae for individual clinical response to DB00544 in the same way using 41 clinical samples obtained from 30 gastric cancer patients and found to be of predictive value in terms of survival , time to treatment failure and tumor growth . None of the 12 selected genes alone could predict such clinical responses . DB04216 protects mouse brain against lead-induced neurotoxicity . DB04216 ( QE ) , the major bioflavonoid in the human diet , has been reported to have many benefits and medicinal properties . However , its protective effects against lead ( Pb ) -induced neurotoxicity have not been clarified . The aim of the present study was to investigate the effects of QE on neurotoxicity in mice exposed to Pb . Mice were exposed to lead acetate ( 20 mg/kg body weight/day ) intragastrically with or without QE ( 15 and 30 mg/kg body weight/day ) coadministration for 3 months . The data showed that QE significantly prevented Pb-induced neurotoxicity in a dose-dependent manner . Exploration of the underlying mechanisms of QE action revealed that QE administration decreased Pb contents in blood ( 13.2 , 19.1 % ) and brain ( 17.1 , 20.0 % ) . QE markedly increased NO production ( Q04695 , 61.1 % ) and PKA activity ( 51.0 , 57.8 % ) in brains of Pb-treated mice . Additionally , QE remarkably suppressed Pb-induced oxidative stress in mouse brain . Western blot analysis showed that QE increased the phosphorylations of Akt , CaMKII P29475 , P29474 , and CREB in brains of Pb-treated mice . The results suggest that QE can inhibit Pb-induced neurotoxicity and partly restore PKA , Akt , NOS , CaMKII , and CREB activities . Pharmacogenetics and future strategies in treating hyperglycaemia in diabetes . This review focuses on current evidence for pharmacogenetics for the 3 commonly used drug classes in treating diabetes : metformin , sulphonylureas and thiazolidinediones . Currently , metformin pharmacogenetics is focussing on drug transport with the recent finding that variation in O75051 transporters might affect metformin response . An aetiological approach has identified monogenic patients with diabetes due to TCF1 mutations who are particularly sensitive to the hypoglycaemic effects of sulphonylureas , and Q14654 or Q09428 mutations in which sulphonylureas can be used in place of insulin treatment . In Type 2 diabetes sulphonylurea response has been shown to be associated with variants Q9NQB0 associated with type 2 diabetes risk . For thiazolidinediones , focus has been on PPARgamma variants although with no consistent result . Genome wide association studies offer great potential to unravel what genetic factors influence response and side effects of diabetes therapies . Large numbers of well phenotyped patients for response and side effect as well as similarly sized similarly phenotyped replication cohorts are required . Establishing such cohorts is a priority in diabetes pharmacogenetics research . Synthetic triterpenoid induces P15428 expression and suppresses inflammation-driven colon carcinogenesis . Colitis-associated colon cancer ( CAC ) develops as a result of inflammation-induced epithelial transformation , which occurs in response to inflammatory cytokine-dependent downregulation of 15-hydroxyprostaglandin dehydrogenase ( P15428 ) and subsequent suppression of prostaglandin metabolism . Agents that both enhance P15428 expression and suppress cyclooxygenase-2 ( P35354 ) production may more effectively prevent CAC . Synthetic triterpenoids are a class of small molecules that suppress P35354 as well as inflammatory cytokine signaling . Here , we found that administration of the synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-C28-methyl ester ( CDDO-Me ) suppresses CAC in mice . In a spontaneous , inflammation-driven intestinal neoplasia model , deletion of Q13485 specifically in T cells led to progressive production of inflammatory cytokines , including P01375 -α , IFN-γ , P35228 , P05231 , IL-1β ; as well as activation of P42224 and P40763 ; along with suppression of P15428 expression . Oral administration of CDDO-Me to mice with Q13485 -deficient T cells increased survival and suppressed intestinal epithelial neoplasia by decreasing production of inflammatory mediators and increasing expression of P15428 . Induction of P15428 by CDDO-Me was dose dependent in epithelial cells and was abrogated following treatment with TGF-β signaling inhibitors in vitro . Furthermore , CDDO-Me-dependent P15428 induction was not observed in P84022 -/- mice . Similarly , CDDO-Me suppressed azoxymethane plus dextran sodium sulfate-induced carcinogenesis in wild-type animals , highlighting the potential of small molecules of the triterpenoid family as effective agents for the chemoprevention of CAC in humans . Mutual exclusivity analysis of genetic and epigenetic drivers in melanoma identifies a link between p14 Q8N726 and RARβ signaling . Melanoma genomes contain thousands of alterations including : mutations , copy number alterations , structural aberrations , and methylation changes . The bulk of this variation is stochastic and functionally neutral , with only a small minority representing " drivers " that contribute to the genesis and maintenance of tumors . Drivers are often directly or inversely correlated across tumors , reflecting the molecular and regulatory signaling pathways in which they operate . Here , a profile of genetic and epigenetic drivers in 110 human melanoma cell lines was generated and searched for non-random distribution patterns . Statistically significant mutual exclusivity was revealed among components of each of the p16(INK4A)- P11802 -RB , DB01367 -RAF-MEK- P29323 and PI3K-AKT signaling pathways . In addition , an inverse correlation was observed between promoter hypermethylation of retinoic acid receptor β ( P10826 ) and CDKN2A alterations affecting p14( Q8N726 ) ( P < 0.0001 ) , suggesting a functional link between RARβ signaling and the melanoma-suppressive activities of p14( Q8N726 ) . Mechanistically , all-trans retinoic acid ( DB00755 ) treatment increased the expression of p14( Q8N726 ) in primary human melanocytes and the steady-state levels of p14( Q8N726 ) in these cells were shown to be regulated via RARβ . Furthermore , the ability of DB00755 to induce senescence is reduced in p14( Q8N726 )-depleted melanocytes , and we provide proof-of-concept that DB00755 can induce irreversible growth arrest in melanoma cells with an intact RARβ-p14( Q8N726 ) signaling axis , independent of p16(INK4A) and p53 status . IMPLICATIONS : These data highlight the power of mutual exclusivity analysis of cancer drivers to unravel molecular pathways and establish a previously unrecognized cross-talk between RARβ and p14( Q8N726 ) with potential implications for melanoma treatment . Effects of P04626 amplicon size and genomic alterations of chromosomes 1 , 3 , and 10 on patient response to trastuzumab in metastatic breast cancer . DB00072 is widely used for advanced breast cancer patients with P04626 -amplified tumors . Nevertheless , over half of these patients do not have an objective response . One reason may be altered expression of genes that might compensate for P04626 inhibition . We previously mapped the gene-rich region of chromosome 17 telomeric to P04626 , and reported considerable variability in the telomeric extent of the P04626 amplicon . Here we examined whether the variable amplicon size may be associated with patient response to trastuzumab . In addition , we looked at associations between response and several signaling pathway-related genes unrelated to the P04626 amplicon , including Q9Y243 , P60484 , P42336 , and P35354 . In 35 patients with P04626 -amplified metastatic breast cancer , with 40 % overall response to trastuzumab , fluorescence in situ hybridization identified the telomeric extent of the P04626 amplicon and the status of the several pathway-related genes . Objective response strongly correlated with the telomeric amplicon size , with 62 % of patients with shorter amplicons responding , compared with only 7 % of patients with longer amplicons ( P = 0.0015 ) . Abnormal copy number of P35354 was marginally associated with objective response ( P = 0.066 ) , while abnormal copy numbers of two reference loci , 1q25 and the chromosome 10 centromere , were significantly associated with response . Pairwise combinations of copy number status of these loci and P04626 amplicon size provided stronger associations and identified a group of patients without responders . These results suggest that patient selection for trastuzumab may be improved by considering P04626 amplicon size and genomic status of the 1q25 , P35354 , and centromere 10 loci . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . A transcriptional block in the P60568 promoter at the -150 AP-1 site in effector CD8+ T cells . Both P01730 + and CD8+ T cells that produce P60568 in response to Ag recognition have been isolated . However , most effector CD8+ T cells recovered after exposure to Ag do not produce sufficient P60568 to sustain growth , and depend on P01730 + T helper cells for this obligate growth factor . P60568 expression in P01730 + T cells is primarily controlled at the level of transcription , but mechanisms restricting P60568 production in CD8+ T cells have not been elucidated . To evaluate transcriptional regulation of the P60568 gene in CD8+ T cells , we stably transfected reporter genes into Ag-specific CD8+ T cell clones . P10747 + CD8(+) T cells unable to transcribe the P60568 gene in response to antigenic stimulation had a block in transactivation of the -150 P10747 response element ( CD28RE ) /AP-1 site of the P60568 promoter , but did transactivate the composite NFAT/AP-1 and O75051 /AP-1 sites , and a consensus AP-1 motif . Mutation of the nonconsensus -150 AP-1 site to a consensus AP-1 site , or insertion of a CD28RE/AP-1 consensus site upstream of the native -150 CD28RE/AP-1 site restored transactivation of the altered promoter . These results suggest that the defect at the -150 site may reflect the absence or inactivity of a required factor rather than repression of the P60568 promoter . Coronavirus 3CLpro proteinase cleavage sites : possible relevance to P49591 virus pathology . BACKGROUND : Despite the passing of more than a year since the first outbreak of Severe Acute Respiratory Syndrome ( P49591 ) , efficient counter-measures are still few and many believe that reappearance of P49591 , or a similar disease caused by a coronavirus , is not unlikely . For other virus families like the picornaviruses it is known that pathology is related to proteolytic cleavage of host proteins by viral proteinases . Furthermore , several studies indicate that virus proliferation can be arrested using specific proteinase inhibitors supporting the belief that proteinases are indeed important during infection . Prompted by this , we set out to analyse and predict cleavage by the coronavirus main proteinase using computational methods . RESULTS : We retrieved sequence data on seven fully sequenced coronaviruses and identified the main 3CL proteinase cleavage sites in polyproteins using alignments . A neural network was trained to recognise the cleavage sites in the genomes obtaining a sensitivity of 87.0 % and a specificity of 99.0 % . Several proteins known to be cleaved by other viruses were submitted to prediction as well as proteins suspected relevant in coronavirus pathology . Cleavage sites were predicted in proteins such as the cystic fibrosis transmembrane conductance regulator ( P13569 ) , transcription factors CREB-RP and O75051 -1 , and components of the ubiquitin pathway . CONCLUSIONS : Our prediction method NetCorona predicts coronavirus cleavage sites with high specificity and several potential cleavage candidates were identified which might be important to elucidate coronavirus pathology . Furthermore , the method might assist in design of proteinase inhibitors for treatment of P49591 and possible future diseases caused by coronaviruses . It is made available for public use at our website : http://www.cbs.dtu.dk/services/NetCorona/ . DeltaEF1 mediates TGF-beta signaling in vascular smooth muscle cell differentiation . Alteration in the differentiated state of smooth muscle cells ( SMCs ) is known to be integral to vascular development and the pathogenesis of vascular disease . However , it is still largely unknown how environmental cues translate into transcriptional control of SMC genes . We found that deltaEF1 is upregulated during SMC differentiation and selectively transactivates the promoters of SMC differentiation marker genes , SM alpha-actin and SM myosin heavy chain ( SM-MHC ) . DeltaEF1 physically interacts with P11831 and P84022 , resulting in a synergistic activation of SM alpha-actin promoter . Chromatin immunoprecipitation assays and knockdown experiments showed that deltaEF1 is involved in the control of the SMC differentiation programs induced by TGF-beta signaling . Overexpression of deltaEF1 inhibited neointima formation and promoted SMC differentiation , whereas heterozygous deltaEF1 knockout mice exhibited exaggerated neointima formation . It thus appears deltaEF1 mediates SMC differentiation via interaction with P11831 and P84022 during development and in vascular disease . Indirubin-3'- ( 2,3 dihydroxypropyl ) -oximether ( E804 ) is a potent modulator of LPS-stimulated macrophage functions . Indirubin is a deep-red bis-indole isomer of indigo blue , both of which are biologically active ingredients in Danggui Longhui Wan , an ancient Chinese herbal tea mixture used to treat neoplasia and chronic inflammation and to enhance detoxification of xenobiotics . Multiple indirubin derivatives have been synthesized and shown to inhibit cyclin-dependent kinases ( CDKs ) and glycogen-synthase kinase ( GSK-3β ) with varying degrees of potency . Several indirubins are also aryl hydrocarbon receptor ( P35869 ) agonists , with P35869 -associated activities covering a wide range of potencies , depending on molecular structure . This study examined the effects of indirubin-3'- ( 2,3 dihydroxypropyl ) -oximether ( E804 ) , a novel indirubin with potent P40763 inhibitory properties , on basal and LPS-inducible activities in murine RAW264.7 macrophages . Using a focused commercial qRT-PCR array platform ( SuperArray® ) , the effects of E804 on expression of a suite of genes associated with stress and toxicity were determined . Most genes up-regulated by LPS treatment were suppressed by E804 ; including LPS-induced expression of pro-inflammatory cytokines and receptors , apoptosis control genes , and oxidative stress response genes . Using qRT-PCR as a follow up to the commercial arrays , E804 treatment suppressed LPS-induced P35354 , P35228 , P05231 and P22301 gene expression , though the effects on P35228 and P35354 protein expression were less dramatic . E804 also inhibited LPS-induced secretion of P05231 and P22301 . Functional endpoints , including P35228 and lysozyme enzymatic activity , phagocytosis of fluorescent latex beads , and intracellular killing of bacteria , were also examined , and in each experimental condition E804 suppressed activities . Collectively , these results indicate that E804 is a potent modulator of pro-inflammatory profiles in LPS-treated macrophages . Uremic Toxins Induce ET-1 Release by Human Proximal Tubule Cells , which Regulates Organic Cation Uptake Time-Dependently . In renal failure , the systemic accumulation of uremic waste products is strongly associated with the development of a chronic inflammatory state . Here , the effect of cationic uremic toxins on the release of inflammatory cytokines and endothelin-1 ( ET-1 ) was investigated in conditionally immortalized proximal tubule epithelial cells ( ciPTEC ) . Additionally , we examined the effects of ET-1 on the cellular uptake mediated by organic cation transporters ( OCTs ) . Exposure of ciPTEC to cationic uremic toxins initiated production of the inflammatory cytokines P05231 ( 117 ± 3 % , p < 0.001 ) , P10145 ( 122 ± 3 % , p < 0.001 ) , and ET-1 ( 134 ± 5 % , p < 0.001 ) . This was accompanied by a down-regulation of O75051 mediated 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide ( ASP+ ) uptake in ciPTEC at 30 min ( 23 ± 4 % , p < 0.001 ) , which restored within 60 min of incubation . Exposure to ET-1 for 24 h increased the ASP+ uptake significantly ( 20 ± 5 % , p < 0.001 ) . These effects could be blocked by BQ-788 , indicating activation of an P24530 -receptor-mediated signaling pathway . Downstream the receptor , P35228 inhibition by ( N(G)-monomethyl-l-arginine ) l-NMMA acetate or aminoguanidine , as well as protein kinase C activation , ameliorated the short-term effects . These results indicate that uremia results in the release of cytokines and ET-1 from human proximal tubule cells , in vitro . Furthermore , ET-1 exposure was found to regulate proximal tubular O75051 transport activity in a differential , time-dependent , fashion . Polymorphisms of genes related to endothelial cells are associated with primary biliary cirrhosis patients of Cretan origin . BACKGROUND : Primary biliary cirrhosis ( P10515 ) is an organ specific autoimmune disease of still unidentified genetic etiology . We have shown that endothelins ( ETs ) , produced by the liver endothelial cells are increased in P10515 and may play a major pathogenetic role . AIMS : To study gene polymorphisms related to the endothelial cells ( P29474 , P10153 -1 genes ) and , to investigate whether the previously reported association of P16410 gene polymorphisms is replicated in a genetically homogeneous Greek population . PATIENTS AND METHODS : Genomic DNA was extracted from 100 P10515 patients ( 83 females , 93 % AMA+ , 74/100 Ludwig stage I-II ) and 158 healthy controls . P29474 , P16410 and ET1 polymorphisms were determined by PCR-RFLPs analysis . RESULTS : Both P29474 intron4 VNTR and P29474 exon7 G894T SNP were significantly associated with increased risk in P10515 . P10153 -11 rs2071942 " A " and rs5370 " T " alleles appeared a tendency for association with disease progression . No association was found between P10515 and the P16410 SNPs analyzed . CONCLUSIONS : We demonstrated that P29474 , a gene related to the liver endothelium function is associated with P10515 . Contrarily , the important in adaptive immunity gene P16410 was not associated with the disease in the homogeneous population analyzed . These results are compatible partially with our previous hypothesis that defects of the liver endothelial system , leading to endothelin overproduction , may be a fundamental early pathogenetic mechanism in P10515 . Lack of endothelial nitric oxide synthase aggravates murine pneumococcal meningitis . DB00435 ( NO ) plays a central role in the pathogenesis of bacterial meningitis . However , the role of NO produced by endothelial NO synthase ( P29474 ) in meningitis is still unclear . We investigated the influence of P29474 depletion on the inflammatory host response , intracranial complications , and outcome in experimental pneumococcal meningitis . Leukocyte accumulation in the cerebrospinal fluid was more pronounced in infected P29474 -deficient mice than in infected wild type mice . This effect could be attributed to an increased expression of P16109 , macrophage inflammatory protein-2 , keratinocyte-derived cytokine , and interleukin ( IL ) -1beta in the brain of infected P29474 -deficient mice . However , no differences in the cerebral expression of intercellular adhesion molecule-1 , tumor necrosis factor-alpha , and P05231 as well as of neuronal NOS and inducible NOS could be detected between infected wild type and mutant mice . In addition to enhanced leukocyte infiltration into the P04141 , meningitis-associated intracranial complications including blood-brain barrier disruption and the rise in intracranial pressure were significantly augmented in infected P29474 -deficient mice . The aggravation of intracranial complications was paralleled by a worsening of the disease , as evidenced by a more pronounced hypothermia , an enhanced weight reduction , and an increased death rate . The current data indicate that P29474 deficiency is detrimental in bacterial meningitis . This effect seems to be related to an increased expression of ( certain ) cytokines/chemokines and adhesion molecules ; thus leading to increased meningeal inflammation and , subsequently , to aggravated intracranial complications . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain . Integration of Q96HU1 kinase signal transduction pathways at the serum response element . The ternary complex factor ( TCF ) subfamily of ETS-domain transcription factors bind with serum response factor ( P11831 ) to the serum response element ( SRE ) and mediate increased gene expression . The TCF protein Elk-1 is phosphorylated by the JNK and P29323 groups of mitogen-activated protein ( Q96HU1 ) kinases causing increased DNA binding , ternary complex formation , and transcriptional activation . Activated SRE-dependent gene expression is induced by JNK in cells treated with interleukin-1 and by P29323 after treatment with phorbol ester . The Elk-1 transcription factor therefore integrates Q96HU1 kinase signaling pathways in vivo to coordinate biological responses to different extracellular stimuli . Expression of retinoic acid receptor beta in dermatofibrosarcoma protuberans . BACKGROUND : P10826 ( RAR beta ) has been shown to act as a tumor suppressor in many solid human tumors . To investigate the putative role of RAR beta in dermatofibrosarcoma protuberans ( DFSP ) , we examined the expression of RAR beta in DFSPs and analyzed the correlation of expression patterns between RAR beta and cyclooxygenase ( P36551 ) -2 as well as clinicopathological variables . METHODS : Using tissue microarray and immunohistochemistry , we evaluated nuclear RAR beta staining and cytoplasm P35354 staining in 53 DFSPs . RESULTS : 48 DFSPs ( 90.58 % ) were immunopositive for RAR beta , while 32 DFSPs ( 60.38 % ) were immunopositive for P35354 . RAR beta staining was significantly inversely correlated with P35354 staining ( p < 0.001 ; r =-0.668 ) . CONCLUSIONS : Our data indicated that RAR beta expressed in DFSPs and correlated with P35354 expression . RAR beta may be a potential therapeutic target for unresectable DFSP cases . Proinflammatory cytokines downregulate gene expression and activity of constitutive nitric oxide synthase in porcine pulmonary artery endothelial cells . We evaluated the effects of cytokines on the catalytic activity and expression of porcine pulmonary artery endothelial cell ( PAEC ) constitutive ( P29474 ) and inducible ( P35228 ) isoforms of nitric oxide synthase ( NOS ) . Exposure of PAEC to the combination of P01579 , P01375 , and P01584 did not alter P35228 activity in cytosolic and membrane fractions but significantly ( p < 0.01 ) reduced P29474 activity in the membrane fraction , but not in the cytosolic fraction , after a 24-h exposure . The cytokine-induced loss of membrane fraction P29474 activity was associated with significant reductions of P29474 mRNA and protein content ( p < 0.01 for both ) . Treatment with the protein synthesis inhibitor , cycloheximide , but not the transcriptional inhibitor actinomycin D prevented cytokine-induced reduction of P29474 mRNA expression . These results suggest that cytokine-induced loss of catalytic activity of P29474 is associated with a reduction in P29474 mRNA and protein mass and that cytokines alter P29474 mRNA stability . Inhibition of protein synthesis prevented reduction of P29474 mRNA by cytokines , suggesting that the mechanism by which cytokines alter P29474 mRNA stability involves protein synthesis . DB04630 stimulates vascular smooth muscle cell proliferation via big mitogen-activated protein kinase 1 activation . The nongenomic effects of aldosterone have been implicated in the pathogenesis of various cardiovascular diseases . DB04630 -induced nongenomic effects are attributable in part to the activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) , a classical mitogen-activated protein ( Q96HU1 ) kinase . Q13164 ( Q13164 ) , a newly identified Q96HU1 kinase , has been shown to be involved in cell proliferation , differentiation , and survival . We examined whether aldosterone stimulates Q13164 -mediated proliferation of cultured rat aortic smooth muscle cells ( RASMCs ) . P08235 ( MR ) expression and localization were evaluated by Western blotting analysis and fluorolabeling methods . P27361 /2 and Q13164 activities were measured by Western blotting analysis with the respective phosphospecific antibodies . Cell proliferation was determined by Alamar Blue colorimetric assay . DB04630 ( 0.1 to 100 nmol/L ) dose-dependently activated Q13164 in RASMCs , with a peak at 30 minutes . To clarify whether aldosterone-induced Q13164 activation is an MR-mediated phenomenon , we examined the effect of eplerenone , a selective MR antagonist , on aldosterone-induced Q13164 activation . DB00700 ( 0.1 to 10 micromol/L ) dose-dependently inhibited aldosterone-induced Q13164 activation in RASMCs . DB04630 also stimulated RASMC proliferation , which was inhibited by eplerenone . DB04630 -mediated phenomena were concluded to be attributable to a nongenomic effect because cycloheximide failed to inhibit aldosterone-induced Q13164 activation . Transfection of dominant-negative Q96HU1 kinase/ P29323 kinase 5 ( Q13163 ) , which is an upstream regulator of Q13164 , partially inhibited aldosterone-induced RASMC proliferation , which was almost completely inhibited by MEK inhibitor PD98059 . In addition to the classical steroid activity , rapid nongenomic effects induced by aldosterone may represent an alternative etiology for vascular diseases such as hypertension . Anti-tumor effect of vitamin A and D on head and neck squamous cell carcinoma . OBJECTIVES : DB00162 and D(3) have a very strong differentiation induction effect . STUDY DESIGN : We examined the anti tumor effect on head and neck squamous cell carcinoma ( HNSCC ) by treatment with several vitamins having strong differentiation induction effects in vitro . METHODS : We used KB cell that an oral floor squamous cell carcinoma , vitamins as all-trans retinoic acid ( DB00755 ) , 4- [ 3,5-bis ( trimethylsilyl ) benzamido ] benzoic acid ( TAC-101 ) , 1alpha,25(OH)(2)D(3) ( calcitriol ) and 22-oxa-1,25-(OH)(2)D(3) ( O75051 ) . We determined receptors of vitamin A and D(3) using RT-PCR . Furthermore , we investigated the proliferation of tumor cells in concentration dependency using [3H]TdR uptake method , apoptosis and apoptosis related factors using TUNEL method and real-time PCR , cell cycle changes using flow cytometry , changing of the sensitivity of using MTT method , cytokine production and the angiogenesis factor using ELISA , by treatment with these vitamins . RESULTS : The deficit of P10826 was found in the KB cell . Each vitamin suppressed the cell proliferation , induced apoptosis , and cell cycle arrest , upregulated sensitivity of the chemotherapeutics drugs and downregulated several angiogenesis factors and an apoptotic factor ; survivin . CONCLUSIONS : These results support the idea that vitamin A , D(3) and their derivatives are useful for preventing and/or treating patients with HNSCC . P04818 genotype-directed chemotherapy for patients with gastric and gastroesophageal junction cancers . BACKGROUND : Retrospective studies indicate associations between TSER ( thymidylate synthase enhancer region ) genotypes and clinical outcomes in patients receiving DB00544 based chemotherapy , but well-controlled prospective validation has been lacking . METHODS : In this phase II study ( NCT00515216 registered through ClinicalTrials.gov , http://clinicaltrials.gov/show/NCT00515216 ) , patients with " good risk " TSER genotypes ( at least one TSER*2 allele ) were treated with FOLFOX chemotherapy to determine whether prospective patient selection can improve overall response rates ( ORR ) in patients with gastric and gastroesophageal junction ( GEJ ) cancers , compared with historical outcomes in unselected patients ( estimated 43 % ) . RESULTS : The ORR in genotype-selected patients was Q04695 % ( 9 partial responses out of 23 evaluable patients , 95 % CI , 22.2 to 59.2 ) , not achieving the primary objective of improving ORR . An encouraging disease control rate ( DCR , consisting of partial responses and stable diseases ) of 95.7 % was noted and patients with homozygous TSER*2 genotype showed better tumor response . CONCLUSIONS : In this first prospective , multi-institutional study in patients with gastric or GEJ cancers , selecting patients with at least one TSER*2 allele did not improve the ORR but led to an encouraging DCR . Further studies are needed to investigate the utility of selecting patients homozygous for the TSER*2 allele and additional genomic markers in improving clinical outcomes for patients with gastric and GEJ cancers . TRIAL REGISTRATION : ClinicalTrials.gov NCT00515216 . Reduction of neuronal and inducible nitric oxide synthase gene expression in patients with cystic fibrosis . As a consequence of diminished nitric oxide synthase ( NOS ) protein concentration , the airway concentration of nitric oxide ( NO ) is reduced in patients with cystic fibrosis ( CF ) . This appears to lead to a reduced elimination of such microorganisms as Pseudomonas aeruginosa . The objective of this study was to analyze whether inducible ( P35228 ) , endothelial ( P29474 ) and neuronal ( P29475 ) NOS are reduced at mRNA level and if so whether this is caused directly by the defective CF transmembrane conductance regulator ( P13569 ) . Nasal polyps from three patients with CF and four otherwise healthy patients were obtained . The expression of the three NOS isoenzymes was quantified using real-time PCR . The P35228 expression was assessed in colon carcinoma cells ( CaCo ) transfected with a normal and a mutated ( DeltaF508 ) P13569 . In CF patients , P35228 mRNA expression was 10-to 20-fold and P29475 gene expression was one-fifth to one-tenth that in control patients ( P < 0.001 ) . In CaCo cells , P35228 gene expression under basal and endotoxin-stimulated conditions did not differ between cells transfected with a mutated P13569 and those transfected with an intact P13569 . This observation suggests that cystic fibrosis is associated with reduced P35228 and P29475 gene expression in nasopharyngeal tissue , possibly disturbing the barrier against infective agents already at the site of entrance . Differential regulation of cystic fibrosis transmembrane conductance regulator by interferon gamma in mast cells and epithelial cells . P13569 ( P13569 ) is a P13569 in epithelial cells ; recently , we identified it in mast cells . Previous work that we confirmed showed that interferon gamma ( IFNgamma ) down-regulated P13569 expression in epithelial cells ( T84 ) , but by contrast , we found that IFNgamma up-regulated P13569 mRNA and protein expression in rat and human mast cells . IFNgamma up-regulation of P13569 in mast cells was inhibited by p38 and extracellular signal-regulated kinase ( P29323 ) kinase inhibitors but not a Janus tyrosine kinase (JAK)2 inhibitor , whereas in T84 cells IFNgamma-mediated down-regulation of P13569 was O60674 -dependent and P29323 - and p38-independent . Furthermore , IFNgamma down-regulation of P13569 in T84 epithelial cells was P42224 -dependent , but up-regulation of P13569 in mast cells was P42224 -independent . Thus , differential regulatory pathways of P13569 expression in mast cells and epithelial cells exist that depend upon either p38/ P29323 or JAK/ P35610 pathways , respectively . Surprisingly , IFNgamma treatment of mast cells inhibited Cl(-) efflux , in contrast to up-regulation of P13569 /mRNA and protein expression . However , down-regulation of Cl(-) flux correlated with IFNgamma-mediated inhibition of mediator secretion . This and other work suggests that the effect of IFNgamma on P13569 expression in mast cells is important for their function . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . Involvement of extracellular signal-regulated kinase module in HIV-mediated P01730 signals controlling activation of nuclear factor-kappa B and AP-1 transcription factors . Although the molecular mechanisms by which the HIV-1 triggers either T cell activation , anergy , or apoptosis remain poorly understood , it is well established that the interaction of HIV-1 envelope glycoproteins with cell surface P01730 delivers signals to the target cell , resulting in activation of transcription factors such as NF-kappa B and AP-1 . In this study , we report the first evidence indicating that kinases MEK-1 ( Q96HU1 kinase/Erk kinase ) and P27361 ( extracellular signal-regulated kinase ) act as intermediates in the cascade of events that regulate NF-kappa B and AP-1 activation upon HIV-1 binding to cell surface P01730 . We found that CEM cells transfected with dominant negative forms of MEK-1 or P27361 do not display NF-kappa B activation after HIV-1 binding to P01730 . In contrast , NF-kappa B activation was observed in these cells after PMA stimulation . Although the different cell lines studied expressed similar amounts of P01730 and p56(lck) , HIV-1 replication and HIV-1-induced apoptosis were slightly delayed in cells expressing dominant negative forms of MEK-1 or P27361 compared with parental CEM cells and cells expressing a constitutively active mutant form of MEK-1 or wild-type P27361 . In light of recently published data , we propose that a positive signal initiated following oligomerization of P01730 by the virus is likely to involve a recruitment of active forms of p56(lck) , P04049 , MEK-1 , and P27361 , before AP-1 and NF-kappa B activation . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . Expression of cyclooxygenase-2 ( P35354 ) in tumour and stroma compartments in cervical cancer : clinical implications . This study aims at investigating the relationship between cyclooxygenase-2 expression in tumour vs stroma inflammatory compartment and its possible clinical role . The study included 99 stage IB-IV cervical cancer patients : immunostaining of tumour tissue sections was performed with rabbit antiserum against cyclooxygenase-2 . CD3 , P01730 , CD8 , CD25 , Mast Cell Tryptase monoclonal antibodies were used to characterise stroma inflammatory cells in nine cervical tumours . An inverse relation was found between cyclooxygenase-2 levels ( cyclooxygenase-2 IDV ) of tumour vs stroma compartment ( r=-0.44 , P < 0.0001 ) . The percentage of cases showing high tumour/stromal cyclooxygenase-2 IDV ratio was significantly higher in patients who did not respond to treatment ( 93.3 % ) with respect to patients with partial ( 60.5 % ) , and complete ( 43.7 % ) response ( P= 0.009 ) . Cases with a high tumour/stroma cyclooxygenase-2 IDV ratio had a shorter overall survival rate than cases with a low tumour/stroma cyclooxygenase-2 IDV ( P < 0.0001 ) . In the multivariate analysis advanced stage and the status of tumour/stroma cyclooxygenase-2 IDV ratio retained an independent negative prognostic role . The proportion of CD3(+) , P01730 (+) , and CD25(+) cells was significantly lower in tumours with high tumour/stroma cyclooxygenase-2 IDV ratio , while a higher percentage of mast cells was detected in tumours showing high tumour/stroma cyclooxygenase-2 IDV ratio . Our study showed the usefulness of assessing cyclooxygenase-2 status both in tumour and stroma compartment in order to identify cervical cancer patients endowed with a very poor chance of response to neoadjuvant therapy and unfavourable prognosis . Novel MEK inhibitor trametinib and other retinoblastoma gene ( RB ) -reactivating agents enhance efficacy of 5-fluorouracil on human colon cancer cells . Chemotherapy for colorectal cancer has become more complicated and diversified with the appearance of molecular-targeting agents . DB00544 ( DB00544 ) has been a mainstay of chemotherapy for colorectal cancer , but it is still unknown whether the combining of DB00544 with novel molecular-targeting agents is effective . P04818 ( TS ) is a direct target of DB00544 , and the low TS level has been generally supposed to sensitize DB00544 's efficacy . We therefore hypothesized that RB-reactivating agents could enhance the efficacy of DB00544 , because the RB-reactivating agents could suppress the function of transcription factor E2F of TS gene promoter . We used three RB-reactivating agents , trametinib/GSK1120212 ( MEK inhibitor ) , fenofibrate ( Q07869 α agonist ) , and LY294002 ( PI3K inhibitor ) , with DB00544 against human colon cancer HT-29 and HCT15 cells . DB08911 induced p15 and p27 expression and reduced cyclin D1 levels in HT-29 cells . Fenofibrate also dephosphorlated P27361 /2 and reduced cyclin D1 levels in HT-29 cells . LY294002 induced p27 expression in HCT15 cells . All three agents caused dephosphorylation of RB protein and P55008 -phase arrest with a reduction of TS expression . As a consequence , the combination of DB00544 with each of the agents resulted in a significant decrease of colony numbers in HT-29 or HCT15 cells . These results suggest " RB-reactivation therapy " using molecular-targeting agents to be a new strategy for DB00544 -based chemotherapy . In particular , we strongly expect trametinib , which was discovered in Japan and was recently submitted to FDA for approval , to be used together with established regimens for colorectal cancer . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . P08235 antagonism in the treatment of chronic central serous chorioretinopathy : a pilot study . PURPOSE : Based on experimental data showing that central serous chorioretinopathy could result from overactivation of mineralocorticoid receptor pathway in choroid vessels , the authors studied eplerenone , a mineralocorticoid receptor antagonist , as a potential treatment for chronic central serous chorioretinopathy . METHODS : This nonrandomized pilot study included 13 patients with central serous chorioretinopathy of at least 4-month duration , treated with 25 mg/day of oral eplerenone for a week followed by 50 mg/day for 1 or 3 months . The primary outcome measure was the changes in central macular thickness recorded by optical coherence tomography , and the secondary outcomes included changes in foveal subretinal fluid ( P11831 ) measured by O75051 , in best-corrected visual acuity ( BCVA ) and the percentage of eyes achieving complete resolution of subretinal fluid during the treatment period . RESULTS : Central macular thickness decreased significantly from 352 ± 139 μm at baseline to 246 ± 113 μm and 189 ± 99 μm at 1 and 3 months under eplerenone treatment ( P < 0.05 and P < 0.01 , respectively ) . At 3 months , the subretinal fluid significantly decreased compared with baseline subretinal fluid ( P < 0.01 ) and best-corrected visual acuity significantly improved compared with baseline best-corrected visual acuity ( P < 0.001 ) . CONCLUSION : DB00700 treatment was associated with a significant reduction in central macular thickness , subretinal fluid level , and an improvement in visual acuity . Randomized controlled trials are needed to confirm these encouraging results .