interacts_with DB06777?
Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . Bile acids activate fibroblast growth factor 19 signaling in human hepatocytes to inhibit cholesterol 7alpha-hydroxylase gene expression . Mouse fibroblast growth factor 15 ( FGF15 ) and human ortholog O95750 have been identified as the bile acid-induced intestinal factors that mediate bile acid feedback inhibition of cholesterol 7alpha-hydroxylase gene ( C YP7A1 ) transcription in mouse liver . The mechanism underlying FGF15/ O95750 inhibition of bile acid synthesis in hepatocytes remains unclear . Chenodeoxycholic acid ( DB06777 ) and the farnesoid X receptor ( Q96RI1 ) -specific agonist GW4064 strongly induced O95750 but inhibited P22680 messenger RNA ( mRNA ) levels in primary human hepatocytes . O95750 strongly and rapidly repressed P22680 but not small heterodimer partner ( Q15466 ) mRNA levels . Kinase inhibition and phosphorylation assays revealed that the mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 ( MAPK/Erk1/2 ) pathway played a major role in mediating O95750 inhibition of P22680 . However , small interfering RNA ( siRNA ) knockdown of Q15466 did not affect O95750 inhibition of P22680 . Interestingly , DB06777 stimulated tyrosine phosphorylation of the FGF receptor 4 ( P22455 ) in hepatocytes . O95750 antibody and siRNA specific to P22455 abrogated GW4064 inhibition of P22680 . These results suggest that bile acid-activated Q96RI1 is able to induce O95750 in hepatocytes to inhibit P22680 by an autocrine/paracrine mechanism . CONCLUSION : The hepatic O95750 / P22455 /Erk1/2 pathway may inhibit P22680 independent of Q15466 . In addition to inducing O95750 in the intestine , bile acids in hepatocytes may activate the liver O95750 / P22455 signaling pathway to inhibit bile acid synthesis and prevent accumulation of toxic bile acid in human livers . Heymann antibodies induce complement-dependent injury of rat glomerular visceral epithelial cells . The aim of this study was to investigate the in vitro role of the complement membrane attack complex ( MAC ) in the injury induced by nephritogenic anti-brush border vesicle ( Fx1A ) antibodies on rat glomerular visceral epithelial cells ( GEC ) . Both sheep and rabbit anti-rat brush border vesicle IgG-induced complement-dependent lysis of cultured GEC . Fab fragments of sheep anti-rat brush border vesicles and polyclonal or monoclonal P98164 IgG were devoid of lytic activity . Shedding of cell-surface antigens induced by sheep or rabbit anti-rat brush border vesicle IgG protected GEC from subsequent exposure to lytic antibodies and complement , an effect that was not obtained with Fab fragments . When GEC were incubated with sheep or rabbit anti-rat brush border vesicle IgG in capping conditions , the P01024 component was co-redistributed with Heymann immune complexes ; in contrast , the MAC remained diffusely bound to the cell surface , indicating that it was not associated with the antigen-antibody complexes . The MAC was demonstrated on the surface of GEC by immunofluorescence staining with anti-MAC neoantigen and by electron microscopy of negatively stained membranes showing focal clusters of 110 A MAC lesions . When GEC were treated with sheep IgG or rabbit IgG plus P13671 -deficient sera , the cells were not lysed and MAC was not demonstrable on the surface ; however , lytic activity was restored when P13671 -deficient sera were reconstituted with purified P13671 . The results are consistent with the interpretation that injury induced by Heymann antibodies on GEC is MAC-dependent . Molecular dissection of dimethylnitrosamine ( O15061 ) -induced hepatotoxicity by mRNA differential display . Differential display reverse transcription-polymerase chain reaction ( DDRT-PCR ) was used to catalogue altered hepatic transcript expression during dimethylnitrosamine ( O15061 ) exposure in vivo . Mice were administered O15061 ( 1.5 or 5 mg/kg ) or vehicle ( phosphate-buffered saline ) i.p. once daily for up to 7 days , and livers were collected 6 h post-injection . Total RNA was reverse transcribed and cDNA subsets were selectively amplified by PCR . DDRT-PCR products were fractionated on denaturing polyacrylamide gels , and differentially expressed bands were excised , reamplified , and subsequently cloned into a plasmid vector . This study identified 23 cDNAs that were induced and 25 cDNAs that were suppressed during O15061 exposure . Altered expression during O15061 exposure for cDNA clones was confirmed by Northern blotting , RNase protection , or in situ hybridization analyses . DNA sequence information indicated that four cDNAs suppressed during O15061 exposure encode cytochrome P450 isoenzyme-cholesterol 7 alpha-hydroxylase ( P22680 ) , a monokine , a myeloid cell differentiation protein , and mouse major urinary protein ( MUP ) . We further observed a O15061 -induced increase in transcripts for complement factor 3 ( P01024 ) and serum amyloid A ( P0DJI8 ) . In contrast , the remaining differentially expressed transcripts detected by DDRT-PCR during O15061 exposure demonstrated no similarity to sequences present in Genbank , suggesting that they may encode previously unreported gene products . In situ hybridization showed MUP transcripts to be expressed by hepatic centrilobular areas that undergo necrosis during subchronic O15061 exposure . Thus , the utilization of DDRT-PCR has identified several differentially expressed hepatic mRNAs associated with various doses and stages of O15061 exposure . Protection of lung epithelial cells from protease-mediated injury by trappin-2 A62L , an engineered inhibitor of neutrophil serine proteases . Neutrophil serine proteases ( NSPs ) , including elastase , proteinase 3 and cathepsin G , play critical roles in the pathogenesis of chronic inflammatory lung diseases . The release of excess NSPs leads to the destruction of lung tissue and an overexuberant , sustained inflammatory response . Antiproteases could be valuable tools for controlling these NSP-mediated inflammatory events . We have examined the capacity of trappin-2 A62L , a potent engineered inhibitor of all three NSPs , to protect human lung A549 epithelial cells from the deleterious effects of NSPs . Trappin-2 A62L , significantly inhibited the detachment of A549 cells and the degradation of the tight-junction proteins , P12830 , β-catenin and ZO-1 , induced by each individual NSP and by activated neutrophils . Trappin-2 A62L also decreased the release of the pro-inflammatory cytokines P05231 and P10145 from A549 cells that had been stimulated with elastase or LPS . Trappin-2 A62D/M63L , a trappin-2 variant that has no antiprotease activity , has similar properties , suggesting that the anti-inflammatory action of trappin-2 is independent of its antiprotease activity . Interestingly , we present evidence that trappin-2 A62L , as well as wild-type trappin-2 , enter A549 cells and move rapidly to the cytoplasm and nucleus , where they are likely to exert their anti-inflammatory effects . We have also demonstrated that trappin-2 A62L inhibits the early apoptosis of A549 cells mediated by NSPs . Thus , our data indicate that trappin-2 A62L is a powerful anti-protease and anti-inflammatory agent that could be used to develop a treatment for patients with inflammatory lung diseases . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Bile acid-induced negative feedback regulation of the human ileal bile acid transporter . Ileal expression of the apical sodium-dependent bile acid transporter ( Q12908 ) in the rat is unaffected by bile salts , yet in the mouse it is under negative-feedback regulation . The bile acid responsiveness of human Q12908 is unknown . The human Q12908 promoter linked to a luciferase reporter was studied in Caco-2 cells treated with chenodeoxycholic acid ( DB06777 ) and transfected with expression plasmids for farnesoid Q9UBH6 ( Q96RI1 ) , short heterodimer partner ( Q15466 ) , and retinoic acid receptor/retinoid X receptor ( RAR/RXR ) . DB06777 treatment of Caco-2 cells led to a 75 % reduction in steady-state Q12908 messenger RNA levels and a 78 % reduction in human Q12908 promoter activity . A dominant negative Q96RI1 abrogated the response to DB06777 . Site-directed mutagenesis of an RAR/RXR cis element in the human Q12908 promoter reduced its activity by 50 % and eliminated the bile acid response . Retinoic acid activated the human Q12908 promoter fourfold . Q15466 repressed the activity of the Q12908 promoter and reduced activation by retinoic acid . Antisense mediated knock-down of Q15466 in Caco-2 cells partially offset the bile acid mediated repression of Q12908 promoter activity . In conclusion , the human Q12908 is positively regulated by retinoic acid . Bile acids induce a negative feedback regulation of human Q12908 via an Q96RI1 -mediated , Q15466 -dependent effect upon RAR/RXR activation of Q12908 . Oleanolic acid is a selective farnesoid X receptor modulator . Oleanolic acid ( OA ) is an ingredient found in some Traditional Chinese Medicine remedies for treating liver ailments . Bile acid biosynthesis and catabolism are in part controlled in the liver by transcription factor farnesoid X receptor ( Q96RI1 ) . It was hypothesized that OA may act through Q96RI1 to mediate some of its beneficial health effects . In this study , it was found that OA bound to the ligand binding domain ( LBD ) of Q96RI1 and blocked its ability to interact with coactivator Q9Y6Q9 . OA also dose-dependently suppressed the activity of Q96RI1 -LBD induced by its endogenous ligand chenodoxycholic acid ( DB06777 ) . Consistently , OA partially blocked the ability of DB06777 to induce a Q96RI1 target gene bile salt export protein ( O95342 ) . On the other hand , OA did not affect the expression of another Q96RI1 target gene organic solute transporter ( Q86UW2 ) . Intriguingly , OA modestly enhanced the expression of a third Q96RI1 target gene short heterodimer partner ( Q15466 ) . This evidence collectively suggested that OA acts as a gene selective modulator of Q96RI1 . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Identification of a transcriptionally active peroxisome proliferator-activated receptor alpha -interacting cofactor complex in rat liver and characterization of Q9BYK8 as a coactivator . Q07869 ( Q07869 alpha ) plays a central role in the cell-specific pleiotropic responses induced by structurally diverse synthetic chemicals designated as peroxisome proliferators . Transcriptional regulation by liganded nuclear receptors involves the participation of cofactors that form multiprotein complexes to achieve cell- and gene-specific transcription . Here we report the identification of such a transcriptionally active Q07869 alpha-interacting cofactor ( PRIC ) complex from rat liver nuclear extracts that interacts with full-length Q07869 alpha in the presence of ciprofibrate , a synthetic ligand , and leukotriene B(4) , a natural ligand . The liganded Q07869 alpha-PRIC complex enhanced transcription from a peroxisomal enoyl- DB01992 hydratase/l-3-hydroxyacyl- DB01992 dehydrogenase bifunctional enzyme gene promoter template that contains peroxisome proliferator response elements . Rat liver PRIC complex comprises some 25 polypeptides , and their identities were established by mass spectrometry and limited sequence analysis . Eighteen of these peptides contain one or more LXXLL motifs necessary for interacting with nuclear receptors . PRIC complex includes known coactivators or coactivator-binding proteins ( CBP , Q15788 , PBP , PRIP , PIMT , O75448 , Q09428 -2 , and P20142 -1 ) , other proteins that have not previously been described in association with transcription complexes ( CHD5 , TOG , and Q8WYB5 ) , and a few novel polypeptides designated PRIC300 , -285 , -215 , -177 , and -145 . We describe the cDNA for Q9BYK8 , which contains five LXXLL motifs . It interacts with Q07869 alpha and acts as a coactivator by moderately stimulating Q07869 alpha-mediated transcription in transfected cells . We conclude that liganded Q07869 alpha recruits a distinctive multiprotein complex from rat liver nuclear extracts . The composition of this complex may provide insight into the basis of tissue and species sensitivity to peroxisome proliferators . Sesquiterpenoids from Atractylodes macrocephala act as farnesoid X receptor and progesterone receptor modulators . Two sesquiterpenoids , atractylenolide II and III , were isolated and identified from Atractylodes macrocephala ( Asteraceae ) to be subsequently evaluated for their activity against farnesoid X receptor ( Q96RI1 ) and progesterone receptor ( PR ) by transient transfection reporter assays . These sesquiterpenoids did not exert significant agonistic effect but antagonized the activity of chenodeoxycholic acid ( DB06777 ) , an endogenous Q96RI1 agonist , for Q96RI1 driven Q15466 promoter transactivation . Additionally , they transactivated P22680 gene promoter activity by antagonizing Q96RI1 . Apart from acting as a Q96RI1 antagonist , atractylenolide III also showed agonistic activity against PR . All these results demonstrated that atractylenolide II and III are the active components of Atractylodes macrocephala to exert specific pharmacologic effects . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Activation of P00533 promotes squamous carcinoma SCC10A cell migration and invasion via inducing EMT-like phenotype change and P14780 -mediated degradation of P12830 . P00533 is a potent stimulator of invasion and metastasis in head and neck squamous cell carcinomas ( HNSCC ) . However , the mechanism by which P00533 may stimulate tumor cell invasion and metastasis still need to be elucidated . In this study , we showed that activation of P00533 by P01133 in HNSCC cell line SCC10A enhanced cell migration and invasion , and induced loss of epitheloid phenotype in parallel with downregulation of P12830 and upregulation of P19022 and vimentin , indicating that P00533 promoted SCC10A cell migration and invasion possibly by an epithelial to mesenchymal transition ( EMT ) -like phenotype change . Interestingly , activation of P00533 by P01133 induced production of matrix metalloproteinase-9 ( P14780 ) and soluble P12830 ( sE-cad ) , and knockdown of P14780 by siRNA inhibited sE-cad production induced by P01133 in SCC10A . Moreover , both P14780 knockdown and P12830 overexpression inhibited cell migration and invasion induced by P01133 in SCC10A . The results indicate that P00533 activation promoted cell migration and invasion through inducing P14780 -mediated degradation of P12830 into sE-cad . Pharmacologic inhibition of P00533 , MEK , and PI3K kinase activity in SCC10A reduced phosphorylated levels of P27361 /2 and AKT , production of P14780 and sE-cad , cell migration and invasion , and expressional changes of EMT markers ( P12830 and P19022 ) induced by P01133 , indicating that P00533 activation promotes cell migration and invasion via P27361 /2 and PI3K-regulated P14780 / P12830 signaling pathways . Taken together , the data suggest that P00533 activation promotes HNSCC SCC10A cell migration and invasion by inducing EMT-like phenotype change and P14780 -mediated degradation of P12830 into sE-cad related to activation of P27361 /2 and PI3K signaling pathways . Feedback regulation of bile acid synthesis in human liver : importance of HNF-4alpha for regulation of P22680 . A great number of nuclear factors are involved in the negative feedback mechanism regulating bile acid synthesis . There are two major ways for the negative feedback to effect the synthesis ; the Q15466 -dependent , involving Q96RI1 , and the Q15466 -independent way , affecting HNF-4alpha . We studied 23 patients with gallstone disease . Eight patients were treated with chenodeoxycholic acid , 7 with cholestyramine prior to operation , and 8 served as controls . Liver biopsies were analyzed with Real-time-PCR . In the cholestyramine-treated group mRNA levels of P22680 were increased about 10-fold . Treatment with DB06777 decreased the mRNA levels of P22680 by about 70 % . The mRNA levels of Q9UNU6 , Q02318 , and O75881 were not significantly altered in the treated groups . The analysis of mRNA levels for HNF-4alpha showed 64 % higher levels in the cholestyramine-treated group compared to the controls . These levels showed positive and highly significant correlation to the levels of mRNA of P22680 when studied in all three groups together . Q96RI1 , Q15466 , and O00482 / O00482 were not significantly affected by the different treatments . Our results indicate that when bile acid synthesis is upregulated by cholestyramine treatment the Q15466 -independent pathway for controlling P22680 transcription dominates over the Q15466 -dependent pathway . MGC9753 gene , located within Q9UD71 - Q14849 - P04626 - Q14451 amplicon on human chromosome 17q12 , encodes the seven-transmembrane receptor with extracellular six-cystein domain . MYC , P04626 , MET , P21802 , P24864 , P04198 , P09544 , P16070 , Q00987 , Q9Y6Q9 , P46940 and O14965 loci are amplified in human gastric cancer . It has been reported that the gene corresponding to EST H16094 is co-amplified with P04626 gene in human gastric cancer . Here , we identified and characterized the gene corresponding to EST H16094 by using bioinformatics . BLAST programs revealed that EST H16094 was derived from the uncharacterized MGC9753 gene . Two ORFs were predicted within human MGC9753 mRNA , and ORF1 ( nucleotide position 18-980 of NM_033419.1 ) was predicted as the coding region of human MGC9753 mRNA based on comparative genomics . Nucleotide sequence of mouse Mgc9753 mRNA was next determined in silico by modification of AK052486 cDNA ( deleting C at the nucleotide position 37 ) . Human MGC9753 and mouse Mgc9753 proteins were 320-amino-acid seven-transmembrane receptors with the N-terminal six-cysteine domain and an N-glycosylation site ( 85.0 % total-amino-acid identity ) . Human MGC9753 protein showed 90.6 % total-amino-acid identity with human CAB2 aberrant protein , which lacked the third-transmembrane domain of MGC9753 due to frame shifts within ORF . Human MGC9753 gene , consisting of eight exons , were clustered with Q9UD71 , Q14849 , O15273 , PNMT , P04626 , MGC14832 and Q14451 genes within the 120-kb region . Q9UD71 , Q14849 , MGC9753 , P04626 and Q14451 genes are co-amplified in several cases of gastric cancer . This is the first report on comprehensive characterization of the amplicon around the Q9UD71 - Q14849 - O15273 -PNMT-MGC9753- P04626 -MGC14832- Q14451 locus on human chromosome 17q12 . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Activation of the farnesoid X receptor represses Q8NBP7 expression in human hepatocytes . The purpose of this study was to determine whether bile acids ( BAs ) modulate hepatic pro-protein convertase subtilisin/kexin 9 ( Q8NBP7 ) gene expression . Immortalized human hepatocytes were treated with various BAs . Chenodeoxycholic acid ( DB06777 ) treatment specifically decreased both Q8NBP7 mRNA and protein contents . Moreover , activation of the BA-activated farnesoid X receptor ( Q96RI1 ) by its synthetic specific agonist GW4064 also decreased Q8NBP7 expression . Of functional relevance , coadministration of DB06777 counteracted the statin-induced Q8NBP7 expression , leading to a potentiation of P01130 activity . This study suggests that a transcriptional repression of Q8NBP7 by DB06777 or Q96RI1 agonists may potentiate the hypolipidemic effect of statins . The unexpected effect of cyclosporin A on CD56+CD16- and CD56+CD16+ natural killer cell subpopulations . DB00091 ( Q13216 ) is commonly used to prevent graft-versus-host disease . The influence of Q13216 on T-cell function has been extensively investigated ; however , the effect of Q13216 on natural killer ( NK ) cells is less understood . NK cells were cultured with P60568 and P40933 with and without Q13216 for 1 week . Compared with controls , Q13216 -treated cultures showed fewer CD56(+)CD16(+) P55040 (+) NK cells and a reciprocal increase in CD56(+)CD16(-) P55040 (-) cells . These changes were due mainly to a reduced proliferation of the CD56(dim) NK-cell subpopulation and a relative resistance of CD56(bright) NK cells to Q13216 . Following coculture with K562 targets , Q13216 -exposed NK cells differed from controls and lacked Ca(2+) oscillations , nuclear factor of activated T cells ( NFAT ) dephosphorylation , and NFAT nuclear translocation . NK cells cultured in Q13216 retained cytotoxicity against K562 , Raji , and P55040 ligand-expressing lymphoblastoid cells . NK cells cultured in Q13216 showed increases in O14931 and reductions in O95944 and P26718 . Following IL-12 and Q14116 stimulation , Q13216 -treated NK cells showed more P01579 -producing cells . Using in vitro NK-cell differentiation , progenitor cells gave rise to more CD56(+) P55040 (-) NK cells in the presence of Q13216 than controls . Collectively , these studies show that Q13216 influences NK-cell function and phenotype , which may have important implications for graft-versus-leukemia effects . All-trans retinoic acid regulates hepatic bile acid homeostasis . Retinoic acid ( RA ) and bile acids share common roles in regulating lipid homeostasis and insulin sensitivity . In addition , the receptor for RA ( retinoid x receptor ) is a permissive partner of the receptor for bile acids , farnesoid x receptor ( Q96RI1 / Q96RI1 ) . Thus , RA can activate the Q96RI1 -mediated pathway as well . The current study was designed to understand the effect of all-trans RA on bile acid homeostasis . Mice were fed an all-trans RA-supplemented diet and the expression of 46 genes that participate in regulating bile acid homeostasis was studied . The data showed that all-trans RA has a profound effect in regulating genes involved in synthesis and transport of bile acids . All-trans RA treatment reduced the gene expression levels of Cyp7a1 , Cyp8b1 , and Akr1d1 , which are involved in bile acid synthesis . All-trans RA also decreased the hepatic mRNA levels of Lrh-1 ( Nr5a2 ) and Hnf4α ( Nr2a1 ) , which positively regulate the gene expression of Cyp7a1 and Cyp8b1 . Moreover , all-trans RA induced the gene expression levels of negative regulators of bile acid synthesis including hepatic Fgfr4 , Fxr , and Shp ( Nr0b2 ) as well as ileal Fgf15 . All-trans RA also decreased the expression of Abcb11 and Slc51b , which have a role in bile acid transport . Consistently , all-trans RA reduced hepatic bile acid levels and the ratio of CA/ DB06777 , as demonstrated by liquid chromatography-mass spectrometry . The data suggest that all-trans RA-induced Q15466 may contribute to the inhibition of P22680 and Q9UNU6 , which in turn reduces bile acid synthesis and affects lipid absorption in the gastrointestinal tract . Genome-wide association study identifies O95150 and Q16633 as susceptibility loci for primary biliary cirrhosis in the Japanese population . For the identification of susceptibility loci for primary biliary cirrhosis ( P10515 ) , a genome-wide association study ( GWAS ) was performed in 963 Japanese individuals ( 487 P10515 cases and 476 healthy controls ) and in a subsequent replication study that included 1,402 other Japanese individuals ( 787 cases and 615 controls ) . In addition to the most significant susceptibility region , human leukocyte antigen ( HLA ) , we identified two significant susceptibility loci , O95150 ( rs4979462 ) and Q16633 ( rs4938534 ) ( combined odds ratio [ OR ] = 1.56 , p = 2.84 × 10(-14) for rs4979462 , and combined OR = 1.39 , p = 2.38 × 10(-8) for rs4938534 ) . Among 21 non-HLA susceptibility loci for P10515 identified in GWASs of individuals of European descent , three loci ( P16871 , Q9UKT9 , and P33681 ) showed significant associations ( combined p = 3.66 × 10(-8) , 3.66 × 10(-9) , and 3.04 × 10(-9) , respectively ) and Q14765 and P19838 loci showed suggestive association with P10515 ( combined p = 1.11 × 10(-6) and 1.42 × 10(-7) , respectively ) in the Japanese population . These observations indicated the existence of ethnic differences in genetic susceptibility loci to P10515 and the importance of P01375 signaling and B cell differentiation for the development of P10515 in individuals of European descent and Japanese individuals . Characterization of lipid metabolism in insulin-sensitive adipocytes differentiated from immortalized human DB05914 . There is a great demand for cell models to study human adipocyte function . Here we describe the adipogenic differentiation of a telomerase-immortalized human mesenchymal stem cell line ( hMSC-Tert ) that maintains numerous features of terminally differentiated adipocytes even after prolonged withdrawal of the peroxisome proliferator activated receptor gamma ( PPARgamma ) agonist rosiglitazone . Differentiated hMSC-Tert developed the characteristic monolocular phenotype of mature adipocytes . The expression of adipocyte specific markers was highly increased during differentiation . Most importantly , the presence of the PPARgamma agonist rosiglitazone was not required for the stable expression of lipoprotein lipase , adipocyte fatty acid binding protein and perilipin on mRNA and protein levels . Q15848 expression was post-transcriptionally down-regulated in the absence of rosiglitazone . P01308 sensitivity as measured by insulin-induced phosphorylation of Akt and S6 ribosomal protein was also independent of rosiglitazone . In addition to commonly used adipogenic markers , we investigated further PPARgamma-stimulated proteins with a role in lipid metabolism . We observed an increase of lipoprotein receptor ( P98155 , Q07954 ) and apolipoprotein E expression during differentiation . Despite this increased expression , the receptor-mediated endocytosis of lipoproteins was decreased in differentiated adipocytes , suggesting that these proteins may have an additional function in adipose tissue beyond lipoprotein uptake . A novel bile acid-activated vitamin D receptor signaling in human hepatocytes . Vitamin D receptor ( P11473 ) is activated by natural ligands , 1alpha , 25-dihydroxy-vitamin D(3) [ 1alpha,25(OH)(2)-D(3) ] and lithocholic acid ( LCA ) . Our previous study shows that P11473 is expressed in human hepatocytes , and P11473 ligands inhibit bile acid synthesis and transcription of the gene encoding cholesterol 7alpha-hydroxylase ( P22680 ) . Primary human hepatocytes were used to study LCA and 1alpha,25(OH)(2)-D(3) activation of P11473 signaling . Confocal immunofluorescent microscopy imaging and immunoblot analysis showed that LCA and 1alpha , 25(OH)(2)-D(3) induced intracellular translocation of P11473 from the cytosol to the nucleus and also plasma membrane where P11473 colocalized with caveolin-1 . P11473 ligands induced tyrosine phosphorylation of c-Src and P11473 and their interaction . Inhibition of c-Src abrogated P11473 ligand-dependent inhibition of P22680 mRNA expression . Kinase assays showed that P11473 ligands specifically activated the c-Raf/ Q02750 /2/extracellular signal-regulated kinase ( P29323 ) 1/2 pathway , which stimulates serine phosphorylation of P11473 and hepatocyte nuclear factor-4alpha , and their interaction . Mammalian two-hybrid assays showed a P11473 ligand-dependent interaction of nuclear receptor corepressor-1 and silencing mediator of retinoid and thyroid with P11473 /retinoid X receptor-alpha ( RXRalpha ) . Chromatin immunoprecipitation assays revealed that an P27361 /2 inhibitor reversed P11473 ligand-induced recruitment of P11473 , RXRalpha , and corepressors to human P22680 promoter . In conclusion , P11473 ligands activate membrane P11473 signaling to activate the Q02750 /2/ P27361 /2 pathway , which stimulates nuclear P11473 /RXRalpha recruitment of corepressors to inhibit P22680 gene transcription in human hepatocytes . This membrane P11473 -signaling pathway may be activated by bile acids to inhibit bile acid synthesis as a rapid response to protect hepatocytes from cholestatic liver injury . Interaction of murine peritoneal leukocytes and mesothelial cells : in vitro model system to survey cellular events on serosal membranes during inflammation . All serosal cavities including peritoneum are lined with a simple squamous mesothelium . Primary culture of murine mesothelial cells has been established to study their cellular interactions with peritoneal leukocytes . The mesothelial character was determined by the cytokeratin and vimentin expression . The mesothelial cells expressed P05362 and P16070 molecules . The expression of P05362 , but not P16070 , was significantly enhanced by the treatment with P01375 ( 100 U/ml ) . We have also investigated possible influence of transforming growth factors , TGF-alpha ( 20 ng/ml ) and TGF-beta ( 2 ng/ml ) , and epidermal growth factor ( 20 ng/ml ) . These factors were not found to modulate P05362 or P16070 expression in vitro . During coculture experiments unstimulated mesothelial cells were almost nonadherent for both resident and elicited peritoneal mononuclear leukocytes for several hours . P01375 or P01133 pretreatment of mesothelial cells greatly enhanced their adhesive affinity to peritoneal mononuclear leukocytes , while TGF-beta pretreatment even reduced the low basal adhesion . Prolonged coculture for 3 weeks resulted in remarkable proliferation and differentiation of both resident and elicited monocytes/macrophages on the mesothelial surface . The stimulation of mesothelial cell culture with P01133 resulted in the macrophage colony-stimulating activity ( M- Q13216 ) production . M- Q13216 was mainly due to P09603 as confirmed with anti P09603 monoclonal antibody ; the residual M- Q13216 was not formed by GM- P04141 . After several passages the mesothelial cells started to produce M- Q13216 spontaneously . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . The bile acid receptor Q96RI1 is a modulator of intestinal innate immunity . The farnesoid X receptor ( Q96RI1 ) is a bile acid-regulated nuclear receptor expressed in enterohepatic tissues . In this study we investigated whether Q96RI1 is expressed by cells of innate immunity and regulates inflammation in animal models of colitis . Acute ( 7 days ) and chronic ( 8 wk ) colitis were induced in wild-type and Q96RI1 (-/-) mice by intrarectal administration of trinitrobenzensulfonic acid or by 7-day administration of 5 % dextran sulfate in drinking water . The results of this experiment demonstrate that Q96RI1 is expressed by and exerts counterregulatory effects on cells of innate immunity . Exposure of LPS-activated macrophages to 6-ethyl chenodeoxycholic acid ( 6E- DB06777 ; INT-747 ) a synthetic Q96RI1 ligand , results in a reciprocal regulation of NF-kappaB dependent-genes ( P01375 , IL-1beta , P05231 , P23219 , P35354 , and P35228 ) and induction of Q15466 , a Q96RI1 -regulated gene . Q96RI1 activation stabilizes the nuclear corepressor NCoR on the NF-kappaB responsive element on the IL-1beta promoter . Colon inflammation in Crohn 's disease patients and in rodent models of colitis is associated with a reduced expression of Q96RI1 mRNA . Using two rodent models of colon inflammation , we show that progression of these immune-mediated disorders is exacerbated in Q96RI1 (-/-) mice ( p < 0.01 ) . In vivo treatment with INT-747 attenuates organ injury and immune cell activation . Q96RI1 activation increased the colon expression of P51161 , Q96RI1 , and Q15466 while reducing IL-1beta , P60568 , P05231 , P01375 , and P01579 mRNA expression and attenuating disease severity . In aggregate , these findings provide evidence that Q96RI1 is an essential component of a network of nuclear receptors that regulate intestinal innate immunity and homeostasis . Involvement of multiple elements in Q96RI1 -mediated transcriptional activation of O95750 . The intestinal endocrine hormone human fibroblast growth factor 19 ( O95750 ) is involved in the regulation of not only hepatic bile acid metabolism but also carbohydrate and lipid metabolism . In the present study , bile acid/farnesoid X receptor ( Q96RI1 ) responsiveness in the O95750 promoter region was investigated by a reporter assay using the human colon carcinoma cell line LS174T . The assay revealed the presence of bile acid/ Q96RI1 -responsive elements in the 5'-flanking region up to 8.8 kb of O95750 . Deletion analysis indicated that regions from -1866 to -1833 , from -1427 to -1353 , and from -75 to +262 were involved in Q96RI1 responsiveness . Four , four , and two consecutive half-sites of nuclear receptors were observed in the three regions , respectively . An electrophoretic mobility shift assay ( EMSA ) and chromatin immunoprecipitation ( ChIP ) assay revealed Q96RI1 /retinoid X receptor α ( RXRα ) heterodimer binding in these three regions . EMSA and reporter assays using mutated constructs indicated that the nuclear receptor Q9Y2I1 , ER2 , and Q30134 motifs in the 5'-flanking region were involved in Q96RI1 responsiveness of O95750 . Lithocholic acid ( LCA ) ( 10 μM ) , chenodeoxycholic acid ( DB06777 ) ( 10 μM ) , or GW4064 ( 0.1 μM ) treatment increased reporter activity in a construct including the three motifs under Q96RI1 -expressing conditions whereas LCA and not DB06777 or GW4064 treatment increased the reporter activity under pregnane X receptor ( O75469 ) -expressing conditions . These results suggest that O95750 is transcriptionally activated through multiple Q96RI1 -responsive elements in the promoter region . P49675 -related lipid transfer domain protein 5 binds primary bile acids . Steroidogenic acute regulatory-related lipid transfer ( START ) domain proteins are involved in the nonvesicular intracellular transport of lipids and sterols . The P49675 ( P49675 and Q14849 ) and Q96DR4 subfamilies ( Q96DR4 -6 ) have an internal cavity large enough to accommodate sterols . To provide a deeper understanding on the structural biology of this domain , the binding of sterols to Q9NSY2 , a member of the Q96DR4 subfamily , was monitored . The SAR by NMR [ (1)H-(15)N heteronuclear single-quantum coherence ( HSQC ) ] approach , complemented by circular dichroism ( CD ) and isothermal titration calorimetry ( ITC ) , was used . Titration of Q9NSY2 with cholic ( CA ) and chenodeoxycholic acid ( DB06777 ) , ligands of the farnesoid X receptor ( Q96RI1 ) , leads to drastic perturbation of the (1)H-(15)N HSQC spectra and the identification of the residues in contact with those ligands . The most perturbed residues in presence of ligands are lining the internal cavity of the protein . Ka values of 1.8·10-(4) M(-1) and 6.3·10(4) M(-1) were measured for CA and DB06777 , respectively . This is the first report of a START domain protein in complex with a sterol ligand . Our original findings indicate that Q9NSY2 may be involved in the transport of bile acids rather than cholesterol . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . Chenodeoxycholic acid-mediated activation of the farnesoid X receptor negatively regulates hydroxysteroid sulfotransferase . Hydroxysteroid sulfotransferase catalyzing bile acid sulfation plays an essential role in protection against lithocholic acid ( LCA ) -induced liver toxicity . Hepatic levels of Sult2a is up to 8-fold higher in farnesoid X receptor-null mice than in the wild-type mice . Thus , the influence of Q96RI1 ligand ( chenodeoxycholic acid ( DB06777 ) and LCA ) feeding on hepatic Sult2a expression was examined in Q96RI1 -null and wild-type mice . Hepatic Sult2a protein content was elevated in Q96RI1 -null and wild-type mice fed a LCA ( 1 % and 0.5 % ) diet . Treatment with 0.5 % DB06777 diet decreased hepatic Sult2a to 20 % of the control in wild-type mice , but increased the content in Q96RI1 -null mice . Liver Sult2a1 ( St2a4 ) mRNA levels were reduced to 26 % in wild-type mice after feeding of a DB06777 diet , while no decrease was observed on Sult2a1 mRNA levels in Q96RI1 -null mice after DB06777 feeding . A significant inverse relationship ( r(2)=0.523 ) was found between hepatic Sult2a protein content and small heterodimer partner ( Q15466 ) mRNA level . Q15149 -mediated increase in Sult2a protein levels were attenuated by DB06777 feeding in wild-type mice , but not in Q96RI1 -null mice . Human Q06520 protein and mRNA levels were decreased in HepG2 cells treated with the Q96RI1 agonists , DB06777 or GW4064 in dose-dependent manners , although Q15466 mRNA levels were increased . These results suggest that SULT2A is negatively regulated through DB06777 -mediated Q96RI1 activation in mice and humans . P01298 enhances plasma glucocorticoid concentration in rats : possible role in hypoglycemic stress . The acute bolus intraperitoneal ( i.p. ) administration of pancreatic polypeptide ( PP ) dose-dependently enhanced the plasma concentration of corticosterone ( P10515 ) in hypophysectomized/ DB01285 replaced rats , but not that of aldosterone . Minimal and maximal effective doses were 10(-12) and 10(-10) mol/rat , respectively , and maximal P10515 increase occurred between 60 and 120 min after PP injection . P01308 ( 1 U/kg , i.p. ) evoked a net decrease in the blood glucose concentration , and marked rises in the plasma levels of PP and P10515 , that attained their maximum at 60 and 120 min , respectively . The effects of insulin were annulled by the simultaneous injection of 0.5 mg/kg atropine . The effects of 1 U/kg insulin and 10(-10) mol/rat PP on P10515 were not additive ; atropine did not affect P10515 response to PP or PP plus insulin , though annulling that to insulin alone . Taken together these findings suggest that PP plays a physiologic role in the rat as modulator of the adrenal response to the insulin-induced hypoglycemic stress . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . P51587 interacts with the cytoskeletal linker protein plectin to form a complex controlling centrosome localization . The breast cancer susceptibility gene ( P51587 ) is localized mainly in the nucleus where it plays an important role in DNA damage repair . Some P51587 protein is also present in the centrosome . Here , we demonstrate that P51587 interacts with plectin , a cytoskeletal cross-linker protein , and that this interaction controls the position of the centrosome . Phosphorylation of plectin by cyclin-dependent kinase 1/cyclin B ( P06493 /CycB ) kinase has been reported to abolish its cross-linking function during mitosis . Here , we induced phosphorylation of plectin in prepared fractions of HeLa cells by adding activated P06493 /CycB kinase . Consequently , there was significant dissociation of the centrosome from the nuclear membrane . Q15149 has six homologous ankyrin-like repeat domains ( termed Q15149 M1-M6 ) . Using a pull-down assay , we found that Q86UG4 - Q15149 M1 and a Q86UG4 -C-terminal region fusion protein ( which comprised Q15149 M6 , along with an adjacent vimentin site ) interacted with P51587 . Since each Q15149 module exhibits high homology to the others , the possibility of all six domains participating in this interaction was indicated . Moreover , when Q15149 M1 was overexpressed in HeLa cells , it competed with endogenous plectin and inhibited the P51587 -plectin interaction . This inhibitory effect resulted in dissociation of the centrosomes from the nucleus and increased the rate of micronuclei formation which may lead to carcinogenesis . In addition , when either P51587 or plectin was suppressed by the appropriate siRNA , a similar change in centrosomal positioning was observed . We suggest that the P51587 -plectin interaction plays an important role in the regulation of centrosome localization and also that displacement of the centrosome may result in genomic instability and cancer development . Chenodeoxycholic acid attenuates ovalbumin-induced airway inflammation in murine model of asthma by inhibiting the T(H)2 cytokines . Asthma is a complex highly prevalent airway disease that is a major public health problem for which current treatment options are inadequate . Recently , farnesoid X receptor ( Q96RI1 ) has been shown to exert anti-inflammatory actions in various disease conditions , but there have been no reported investigations of Chenodeoxycholic acid ( DB06777 ) , a natural Q96RI1 agonist , in allergic airway inflammation . To test the DB06777 effectiveness in airway inflammation , ovalbumin ( OVA ) -induced acute murine asthma model was established . We found that lung tissue express Q96RI1 and DB06777 administration reduced the severity of the murine allergic airway disease as assessed by pathological and molecular markers associated with the disease . DB06777 treatment resulted in fewer infiltrations of cells into the airspace and peribronchial areas , and decreased goblet cell hyperplasia , mucus secretion and serum IgE levels which was increased in mice with OVA-induced allergic asthma . The DB06777 treatment further blocked the secretion of TH2 cytokines ( P05112 , P05113 and P35225 ) and proinflammatory cytokine P01375 -α indicate that the Q96RI1 and its agonists may have potential for treating allergic asthma . Farnesoid X receptor activation by chenodeoxycholic acid induces detoxifying enzymes through AMP-activated protein kinase and extracellular signal-regulated kinase 1/2-mediated phosphorylation of CCAAT/enhancer binding protein β . Farnesoid X receptor ( Q96RI1 ) regulates redox homeostasis and elicits a cytoprotective effect . CCAAT/enhancer binding protein-β ( C/EBPβ ) plays a role in regulating the expression of hepatocyte-specific genes and contributes to hepatocyte protection and liver regeneration . In view of the role of Q96RI1 in xenobiotic metabolism and hepatocyte survival , this study investigated the potential of Q96RI1 to activate C/EBPβ for the induction of detoxifying enzymes and the responsible regulatory pathway . Chenodeoxycholic acid ( DB06777 ) , a major component in bile acids , activates Q96RI1 . In HepG2 cells , DB06777 treatment activated C/EBPβ , as shown by increases in its phosphorylation , nuclear accumulation , and expression . 3-(2,6-Dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl-)oxymethyl-5-isopropyl-isoxazole ( GW4064 ) , a synthetic Q96RI1 ligand , had similar effects . In addition , DB06777 enhanced luciferase gene transcription from the construct containing -1.65-kb P09210 promoter , which contained C/EBP response element ( pGL-1651 ) . Moreover , DB06777 treatment activated AMP-activated protein kinase ( AMPK ) , which led to extracellular signal-regulated kinase 1/2 ( P27361 /2 ) activation , as evidenced by the results of experiments using a dominant-negative mutant of AMPKα and chemical inhibitor . The activation of P27361 /2 was responsible for the activating phosphorylation of C/EBPβ . Q96RI1 knockdown attenuated the ability of DB06777 to activate AMPK and P27361 /2 and phosphorylate C/EBPβ . Consistently , enforced expression of Q96RI1 promoted the phosphorylation of AMPKα , P27361 /2 , and C/EBPβ , verifying that C/EBPβ phosphorylation elicited by DB06777 results from the activation of AMPK and P27361 /2 by Q96RI1 . In mice , DB06777 treatment activated C/EBPβ with the induction of detoxifying enzymes in the liver . Our results demonstrate that DB06777 induces antioxidant and xenobiotic-metabolizing enzymes by activating C/EBPβ through AMPK-dependent P27361 /2 pathway downstream of Q96RI1 . Activation of farnesoid X receptor attenuates liver injury in systemic lupus erythematosus . To investigate the expression and effect of farnesoid X receptor ( Q96RI1 ) on systemic lupus erythematosus ( SLE ) liver dysfunction and indicate its hepatoprotective role and the immunomodulatory property . mRNA and protein levels of Q96RI1 were determined on the liver specimens of SLE patients with liver injury as well as MRL/lpr rodent models . The Q96RI1 agonist chenodeoxycholic acid ( DB06777 ) was administrated to MRL/lpr mice and the control BALB/C with concanavalin A ( ConA ) -induced liver injury . Blood samples were taken 0 , 4 , 8 , 12 , 16 , and 24 h after ConA injection for the detection of serum ALT , Q9NRA2 , IFN-γ , P01375 -α , and P05231 . Q96RI1 was down-regulated at both mRNA and protein levels in the liver specimens of SLE patients with liver injury as well as MRL/lpr mice . MRL/lpr was more susceptible to ConA than BALB/C indicated by significantly higher levels of aminotransferase and inflammatory cytokines . Activation of Q96RI1 by DB06777 significantly reduced aminotransferase and inflammatory cytokines IFN-γ , P01375 -α , and P05231 caused by ConA injection in MRL/lpr mice . Q96RI1 was down-regulated in SLE patients as well as MRL/lpr lupus models with liver dysfunction . Q96RI1 activation ameliorated liver injury and suppressed inflammatory cytokines , thereby showing its protective function in SLE . Our findings raised the promising potential target for the treatment of SLE liver injury . Preparation of phosphorylated starch by dry-heating in the presence of pyrophosphate and its calcium-phosphate solubilizing ability . Starch was phosphorylated through dry-heating in the presence of pyrophosphate at various conditions , and the characteristics of phosphorylated starch ( PS ) were examined . Starch phosphorylation increases as the pH increases from 3 to 6 , but diminishes at pH 7 . Increased temperatures enhance phosphorylation . Data from (31)P NMR suggests that starch phosphorylation occurs mainly at the P01024 -OH and P13671 -OH of the glucose residue . The phosphate linkage is mainly due to monostarch monophosphate . Although starch had almost no calcium phosphate-solubilising capacity , this capacity was markedly enhanced by phosphorylation . X-ray diffraction analysis indicates that the crystal structure of hydroxyapatite was not present in the calcium phosphate-PS complex . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Use of RNA interference to elucidate the effect of P04198 on cell cycle in neuroblastoma . BACKGROUND : P04198 amplification marks poor prognosis in neuroblastoma ( NB ) tumors . In evaluating the mechanisms by which retinoic acid ( RA ) or nerve growth factor ( P01138 ) decrease cell number in P04198 amplified NB cells , we have identified a number of proteins whose expression either decreases ( E2F , P06493 , Q00534 , cyclin dependent kinase activity ) or increases ( p27 ) in association with a decrease in P04198 expression . However , it was still unclear which were P04198 dependent effects or not . PROCEDURE : This study aimed to determine which changes in cell cycle gene expression are modulated as a consequence of the decrease in P04198 . We silenced P04198 expression using siRNA targeted to the coding region of P04198 . Then , by using siRNA transient transfections , we analyzed the change of cell cycle related genes and cell cycle in P04198 amplified NB cell lines . RESULTS : We demonstrate that expression of P04198 can be suppressed by almost 60 % in P04198 amplified NB cell using siRNAs targeted to P04198 . Functionally , the decrease in P04198 leads to a decrease in cells in the S-phase of the cell cycle . Decreases in P04198 are associated with decreases in Q01094 -2 and Q02363 along with increases in p27 protein levels by post-transcriptional modification . Moreover , we find that a decrease in P04198 is accompanied by a decrease in cdk6 mRNA and protein expression . CONCLUSIONS : These results show that E2F and Q02363 expression is associated with P04198 regulation and that cdk6 is a possible new transcriptional target of P04198 . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . The P04626 amplicon in breast cancer : Topoisomerase IIA and beyond . P04626 gene amplification is observed in about 15 % of breast cancers . The subgroup of P04626 -positive breast cancers appears to be heterogeneous and presents complex patterns of gene amplification at the locus on chromosome 17q12-21 . The molecular variations within the chromosome 17q amplicon and their clinical implications remain largely unknown . Besides the well-known P11388 gene encoding Topoisomerase IIA , other genes might also be amplified and could play functional roles in breast cancer development and progression . This review will focus on the current knowledge concerning the P04626 amplicon heterogeneity , its clinical and biological impact and the pitfalls associated with the evaluation of gene amplifications at this locus , with particular attention to P11388 and the link between P11388 and anthracycline benefit . In addition it will discuss the clinical and biological implications of the amplification of ten other genes at this locus ( MED1 , Q14849 , Q14451 , P10827 , P10276 , IGFPB4 , P32248 , P35900 , P08727 and P01350 ) in breast cancer . Overexpression of Q14849 in human monocyte/macrophages induces an anti-atherogenic lipid phenotype . Dysregulated macrophage cholesterol homoeostasis lies at the heart of early and developing atheroma , and removal of excess cholesterol from macrophage foam cells , by efficient transport mechanisms , is central to stabilization and regression of atherosclerotic lesions . The present study demonstrates that transient overexpression of Q14849 { START [ P49675 ( steroidogenic acute regulatory protein ) -related lipid transfer ] domain 3 ; also known as Q14849 ( metastatic lymph node 64 ) } , an endosomal cholesterol transporter and member of the ' START ' family of lipid trafficking proteins , induces significant increases in macrophage O95477 ( DB00171 -binding cassette transporter A1 ) mRNA and protein , enhances [(3)H]cholesterol efflux to apo ( apolipoprotein ) AI , and reduces biosynthesis of cholesterol , cholesteryl ester , fatty acids , triacylglycerol and phospholipids from [(14)C]acetate , compared with controls . Notably , overexpression of Q14849 prevents increases in cholesterol esterification in response to acetylated LDL ( low-density lipoprotein ) , blocking cholesteryl ester deposition . Thus enhanced endosomal trafficking via Q14849 induces an anti-atherogenic macrophage lipid phenotype , positing a potentially therapeutic strategy . A maternal high-fat diet modulates fetal Q96EB6 histone and protein deacetylase activity in nonhuman primates . In nonhuman primates , we previously demonstrated that a maternal high-fat diet ( MHFD ) induces fetal nonalcoholic fatty liver disease ( NAFLD ) and alters the fetal metabolome . These changes are accompanied by altered acetylation of histone H3 ( H3K14ac ) . However , the mechanism behind this alteration in acetylation remains unknown . As Q96EB6 is both a lysine deacetylase and a crucial sensor of cellular metabolism , we hypothesized that Q96EB6 may be involved in fetal epigenomic alterations . Here we show that in utero exposure to a MHFD , but not maternal obesity per se , increases fetal H3K14ac with concomitant decreased Q96EB6 expression and diminished in vitro protein and histone deacetylase activity . MHFD increased H3K14ac and DBC1- Q96EB6 complex formation in fetal livers , both of which were abrogated with diet reversal despite persistent maternal obesity . Moreover , MHFD was associated with altered expression of known downstream effectors deregulated in NAFLD and modulated by Q96EB6 ( e.g. , PPARΑ , P37231 , P36956 , P22680 , P49327 , and O00767 ) . Finally , ex vivo purified Q96EB6 retains deacetylase activity on an H3K14ac peptide substrate with preferential activity toward acetylated histone H3 ; mutagenesis of the catalytic domain of Q96EB6 ( H363Y ) abrogates H3K14ac deacetylation . Our data implicate Q96EB6 as a likely molecular mediator of the fetal epigenome and metabolome under MHFD conditions . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . The nuclear hormone receptor farnesoid X receptor ( Q96RI1 ) is activated by androsterone . Farnesoid X receptor ( Q96RI1 ) uses bile acids as endogenous ligands . Here , we demonstrate that androsterone , a metabolic product of testosterone , is also an Q96RI1 ligand . Treatment of castrated male mice with androsterone induced expression of the Q96RI1 target gene small heterodimer partner ( Q15466 ) . In mouse AML-12 hepatocytes , chenodeoxycholic acid ( DB06777 ) or androsterone induced Q15466 expression with a similar kinetic pattern . The Q96RI1 antagonist guggulsterone blocked the induction of Q15466 by androsterone in AML-12 cells . Nuclear magnetic resonance spectroscopy demonstrated the direct binding of androsterone to purified human Q96RI1 ( hFXR ) ligand-binding domain ( LBD ) protein , resulting in the recruitment of steroid receptor coactivator protein-1 ( Q15788 ) coactivator peptide . In HEK293 cells , androsterone activated gal4-mouse Q96RI1 -LBD and gal4-hFXR-LBD fusion proteins , although in contrast to DB06777 , androsterone activation was significantly greater for the mouse Q96RI1 -LBD than for the hFXR-LBD . Site-directed mutagenesis of the hFXR-LBD defined amino acids Asn354 and Ser345 as critical for differential species sensitivity to DB06777 and androsterone , respectively . Crystal structure studies suggest that the orientation of the steroid nucleus of bile acids within the binding pocket of Q96RI1 is reversed from all other nuclear hormone receptors . In support of this model , we show here that mutations M265I or R331H , residues predicted by crystal structure to interact with the carboxylic acid tail of DB06777 but not with androsterone , altered DB06777 activation but had no effect on androsterone activation . Activation of Q96RI1 by androsterone may provide an additional means for physiological or pharmacological modulation of Q96RI1 . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Benzyl isothiocyanate ( BITC ) inhibits migration and invasion of human gastric cancer AGS cells via suppressing P29323 signal pathways . Metastasis suppressors and associated other regulators of cell motility play a critical initial role in tumor invasion and metastases . Benzyl isothiocyanate ( BITC ) is a hydrolysis compound of glucotropaeolin in dietary cruciferous vegetables . BITC has been found to exhibit prevention of cancers in laboratory animals and might also be chemoprotective in humans . Here , the purpose of this study was to investigate the effects of BITC on cell proliferation , migration , invasion and mitogen-activated protein kinase ( MAPK ) pathways of AGS human gastric cancer cells . Wound healing and Boyden chamber ( migration and invasion ) assays demonstrated that BITC exhibited an inhibitory effect on the abilities of migration and invasion in AGS cancer cells . BITC suppressed cell migration and invasion of AGS cells in a dose-dependent manner . Results from Western blotting indicated that BITC exerted an inhibitory effect on the P27361 /2 , Ras , P62993 , Rho A , P35228 , P35354 for causing the inhibitions of P08253 , -7 and -9 then followed by the inhibitions of invasion and migration of AGS cells in vitro . BITC also promoted O14733 , Q99759 , c-jun , P45983 /2 , P15692 , Sos1 , phosphoinositide 3-kinase ( PI3K ) , PKC , nuclear factor-kappaB ( NF-κB ) p65 in AGS cells . Results from real-time polymerized chain reaction ( PCR ) showed that BITC inhibited the gene expressions of P08253 ,-7 -9 , Q05397 , Q13464 and RhoA after BITC treatment for 24 and 48 hours in AGS cells . Taken together , the finding may provide new mechanisms and functions of BITC , which inhibit migration and invasion of human gastric cancer AGS cells . Serum concentrations of fibroblast growth factor 19 in patients with obesity and type 2 diabetes mellitus : the influence of acute hyperinsulinemia , very-low calorie diet and Q07869 -α agonist treatment . The aim of our study was to measure serum concentrations of fibroblast growth factor 19 ( O95750 ) in patients with obesity ( OB ) , obesity and type 2 diabetes mellitus ( T2DM ) and healthy subjects ( C ) at baseline and after selected interventions . We measured serum O95750 levels and other biochemical and hormonal parameters in 29 OB and 19 T2DM females and 30 sex- and age-matched control subjects . The interventions were acute hyperinsulinemia during isoglycemic-hyperinsulinemic clamp ( n=11 for T2DM and 10 for C ) , very-low calorie diet ( VLCD , n=12 for OB ) and 3 months treatment with Q07869 agonist fenofibrate ( n=11 for T2DM ) . Baseline serum O95750 levels were significantly lower in OB relative to C group ( 132.1+/-12.7 vs. 202.2+/-16.7 pg/ml , p < 0.05 ) , while no significant difference was observed between T2DM and OB or control group . Acute hyperinsulinemia tended to decrease O95750 levels in both healthy and T2DM subjects . Three weeks of VLCD in OB group had no significant effect on O95750 , whereas three months of fenofibrate treatment markedly reduced O95750 levels in T2DM patients ( 194.58+/-26.2 vs. 107.47+/-25.0 pg/ml , p < 0.05 ) . We conclude that O95750 levels in our study were at least partially dependent upon nutritional status , but were not related to parameters of glucose metabolism or insulin sensitivity . 1,25-dihydroxyvitamin D3 and its receptor inhibit the chenodeoxycholic acid-dependent transactivation by farnesoid X receptor . Farnesoid X receptor ( Q96RI1 ) , the receptor for bile acids , including chenodeoxycholic acid ( DB06777 ) , is a member of the nuclear receptor superfamily , which also includes the receptors for retinoic acid , vitamin D ( D3 ) , thyroid hormone , thiazolidinedione and 22(R)-hydroxycholesterol . Here , we have evaluated the effects of a series of ligands and their receptors on the promoter activity induced by DB06777 / Q96RI1 . The kidney cell line , CV1 , was cotransfected with Q96RI1 -expression plasmid and the luciferase-based reporter gene that has a thymidine kinase promoter fused to the canonical Q96RI1 -responsive element or the natural promoter for the small heterodimer partner ( Q15466 ) , bile salt export pump ( O95342 ) , and ileum bile acid ( P51161 ) gene . D3 and its receptor ( P11473 ) inhibited the transactivation of all four reporter constructs that are enhanced by DB06777 / Q96RI1 . The effect of D3 on the expression of the O95342 and Q15466 genes in HepG2 cells and that of the P51161 gene in Caco-2 cells were confirmed by reverse transcription ( RT ) -PCR . Deletion analysis of P11473 revealed that its ligand-binding domain ( LBD ) is responsible for the repression and the DNA-binding domain ( DBD ) is dispensable . Specific interaction between Q96RI1 and P11473 was detected with the in vitro pull-down assay using chimeric Q96RI1 or P11473 fused to glutathione-S-transferase . Farnesoid X receptor responds to bile acids and represses cholesterol 7alpha-hydroxylase gene ( P22680 ) transcription . DB04540 7alpha-hydroxylase gene ( P22680 ) transcription is repressed by bile acids . The goal of this study is to elucidate the mechanism of P22680 transcription by bile acid-activated farnesoid X receptor ( Q96RI1 ) in its native promoter and cellular context and to identify Q96RI1 response elements in the gene . In Chinese hamster ovary cells transfected with retinoid X receptor alpha ( RXRalpha ) / Q96RI1 , only chenodeoxycholic acid ( DB06777 ) and deoxycholic acid ( DB08809 ) were able to stimulate a heterologous promoter/reporter containing an ecdysone response element . In HepG2 cells , all bile acids ( 25 microM ) were able to repress P22680 /luciferase reporter activity , and only DB06777 and DB08809 further repressed reporter activity when cotransfected with RXRalpha/ Q96RI1 . The concentration of DB06777 required to inhibit 50 % of reporter activity ( IC(50) ) was determined to be approximately 25 microM without Q96RI1 and 10 microM with Q96RI1 . Deletion analysis revealed that the bile acid response element located between nucleotides -148 and -128 was the Q96RI1 response element , but RXRalpha/ Q96RI1 did not bind to this sequence . These results suggest that bile acid-activated Q96RI1 exerts its inhibitory effect on P22680 transcription by an indirect mechanism , in contrast to the stimulation and binding of Q96RI1 to intestinal bile acid-binding protein gene promoter . Results also reveal that bile acid receptors other than Q96RI1 are present in HepG2 cells . [ Expression and regulation of megalin in gallbladder mucosa associated with cholesterol gallstone disease ] . OBJECTIVE : To explore the relationship between expression and regulation of P98164 in gallbladder mucosa and cholesterol gallstone disease . METHODS : Gallbladder mucosa , gallbladder wall , bile , gallstone were collected from 29 patients with cholesterol gallstone disease ( GS ) and 12 patients with gallstone free ( P52945 ) . Lipids of bile and stone were measured by kits . Expression of P98164 and O60494 was analyzed by Real-time PCR . GBC-SD cell line were treated with T0901317 , 9-cis retinoic acid , chenodeoxycholic acid ( DB06777 ) , the agonists of LXR , RXR , Q96RI1 , respectively . Gene expressions were detected . RESULTS : Biliary cholesterol % molar and CSI increased significantly in GS group [ ( 7.98 +/- 0.44 ) mol % vs ( 4.87 +/- 0.39 ) mol % , P < 0.01 ] . P98164 expression in GS group was significantly higher than that in P52945 group ( P < 0.05 ) and cubilin expression was similar between two groups . In vitro experiments showed that DB06777 markedly increased expression of megalin . CONCLUSION : This study shows that the increased expression P98164 may help to increase cholesterol uptake in gallbladder and play a compensative role in GS . Q96RI1 may participate in the transcription regulating of P98164 . Farnesoid X receptor protects liver cells from apoptosis induced by serum deprivation in vitro and fasting in vivo . The farnesoid X receptor ( Q96RI1 ) is a key metabolic regulator in the liver by maintaining the homeostasis of liver metabolites . Recent findings suggest that Q96RI1 may have a much broader function in liver physiology and pathology . In the present work , we identify a novel role of Q96RI1 in protecting liver cell from apoptosis induced by nutritional withdrawal including serum deprivation in vitro or starvation in vivo . Two Q96RI1 ligands , chenodeoxycholic acid ( DB06777 ) and GW4064 , rescued HepG2 cells from serum deprivation-induced apoptosis in a dose-dependent manner . This effect of Q96RI1 on apoptotic suppression was compromised when Q96RI1 was knocked down by short interfering RNA . Similarly , the effects of both DB06777 and GW4064 were abolished after inhibition of the MAPK pathway by a specific inhibitor of MAPK kinase 1/2 . Immunoblotting results indicated that Q96RI1 activation by DB06777 and GW4064 induced P27361 /2 phosphorylation , which was attenuated by serum deprivation . In vivo , Q96RI1 (-/-) mice exhibited an exacerbated liver apoptosis and lower levels of phosphorylated- P27361 /2 compared to wild-type mice after starvation . In conclusion , our results suggest a novel role of Q96RI1 in modulating liver cell apoptosis . NO modulates the molecular basis of rat interscapular brown adipose tissue thermogenesis . Molecular mechanisms underlying interscapular brown adipose tissue ( Q12908 ) thermogenesis were elucidated . Namely , gene and/or protein expression of uncoupling protein 1 ( P25874 ) , peroxisome proliferator-activated receptor gamma ( PPARgamma ) , PPARgamma-coactivator-1alpha ( P20142 -1alpha ) , vascular endothelial growth factor ( P15692 ) and proliferating cell nuclear antigen ( P12004 ) - key molecules that regulate thermogenesis-related processes - mitochondriogenesis , angiogenesis and Q12908 hyperplasia , in rats subjected to cold ( 4+/-1 degrees C ) for 1 , 3 , 7 , 12 , 21 and 45days were investigated . Particularly , to examine influence of nitric oxide ( NO ) on Q12908 thermogenic-program , cold-exposed animals were treated by l-arginine or N(omega)-nitro-l-arginine-methyl ester ( L-NAME ) . Related to control ( 22+/-1 degrees C ) , cold induced time-coordinated P25874 , PPARgamma and P20142 -1alpha transcriptional activation accompanied by P12004 activation and increased P15692 immunolabeling that correlate with endothelial NO synthase ( P29474 ) transcriptional activation suggesting NO involvement in these thermogenic-factors activation . Observed molecular changes were translated into increased mitochondrial-remodeling , angiogenesis , and Q12908 hyperplasia . l- DB00125 augmented and prolonged cold-induced increase of P29474 , inducible NOS and thermogenic-molecules expression , Q12908 nerve supply , vascularity , hyperplasia and mitochondrial-remodeling , while L-NAME had an opposite effects . Results show that NO improves thermogenesis-related mitochondriogenesis , angiogenesis and tissue hyperplasia , positively affecting molecular basis of these processes , suggesting that NO is an essential regulator of Q12908 thermogenic-program operating , at genes , proteins and tissue structure levels . O95750 promotes progression of prostate cancer . BACKGROUND : Fibroblast growth factor ( FGF ) signaling pathways have been reported to play important roles in prostate cancer ( PCa ) progression . O95750 is one of a subfamily of FGFs that circulate in serum and act in an endocrine manner . Our objective was to investigate its role in the progression of PCa . METHODS : The effect of O95750 on the proliferation and epithelial-mesenchymal transition of LNCaP and PC3 cells was examined using MTT assay and Western blotting . Serum concentration of O95750 was measured by ELISA in 209 patients with PCa , and the association between clinicopathological features and the presence of O95750 -positive cells in tissues derived from 155 patients who undergone radical prostatectomy was investigated . RESULTS : Under androgen-deprived conditions achieved by incubation in medium with O95750 , the expression of P19022 in LNCaP cells was enhanced , that of P12830 and caspase 3 was suppressed , and the viability of LNCaP and PC3 cells was significantly enhanced . Significantly higher levels of PSA were recorded in the group determined by immunohistochemistry staining to be O95750 -positive ( P = 0.0046 ) . The 5-year biochemical recurrence-free survival rate after radical prostatectomy was 46.4 % in the O95750 -positive group and 70.0 % in the O95750 -negative group ( P = 0.0027 ) . In multivariate analysis , the presence of O95750 -positive tissues was an independent factor for worse prognosis after radical prostatectomy ( P = 0.0052 ) . Serum O95750 levels in high Gleason grade group were higher than that in low Gleason grade group ( P = 0.0009 ) . CONCLUSIONS : O95750 might be associated with biochemical recurrence after radical prostatectomy by promoting cell proliferation and epithelial-mesenchymal transition of PCa . The regulation of sterol metabolism by cell interactions . Total and free cholesterol levels in P13671 glial cells are regulated by a cell interaction-dependent mechanism that operates independently of exogenous cholesterol and serum lipoproteins . This mechanism , which is activated by changes in culture density , coordinately regulates the activities of P04035 and acyl- DB01992 :cholesterol acyltransferase ( ACAT ) . Both enzyme activities are low in sparse density cultures , rise as density increases from sparse to moderate , and decrease with further density increases . When culture density is abruptly elevated , both enzyme activities decay rapidly and with biphasic kinetics . Neither enzyme phosphorylation nor diffusible cytosolic factors appear to be directly involved in density suppression of P04035 . Studies with human fibroblasts that are defective in P01130 function demonstrate that density regulation does not require a functional P01130 . Extracellular matrix and soluble factors have also been ruled out as intercellular mediators . The specific growth rate of P13671 cultures changes with density in the same manner as sterol metabolism . The possibility that growth and sterol metabolism are regulated by a common cell interaction-dependent mechanism is discussed . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . The farnesoid X receptor induces very low density lipoprotein receptor gene expression . The farnesoid X receptor ( Q96RI1 ) is a nuclear receptor activated by bile acids ( BAs ) . In response to ligand-binding , Q96RI1 regulates many genes involved in BA , lipid , and lipoprotein metabolism . To identify new Q96RI1 target genes , microarray technology was used to profile total RNA extracted from HepG2 cells treated with the natural Q96RI1 agonist chenodeoxycholic acid ( DB06777 ) . Interestingly , a significant increase of transcript level of the very low density lipoprotein receptor ( P98155 ) was observed . Our data , resulting from selective Q96RI1 activation , Q96RI1 RNA silencing and Q96RI1 -deficient mice , clearly demonstrate that BAs up-regulate P98155 transcript levels via a Q96RI1 -dependent mechanism in vitro in human and in vivo in mouse liver cells . Histone acetyltransferase-dependent chromatin remodeling and the vascular clock . Rhythmic gene expression is central to the circadian control of physiology in mammals . Transcriptional activation of Per and Cry genes by heterodimeric bHLH- DB00233 proteins is a key event in the feedback loop that drives rhythmicity ; however , the mechanism is not clearly understood . Here we show the transcriptional coactivators and histone acetyltransferases , p300/CBP , Q92831 , and Q9Y6Q9 associate with the bHLH- DB00233 proteins , O15516 and Q99743 , to regulate positively clock gene expression . Furthermore , Cry2 mediated repression of Q99743 : O00327 is overcome by overexpression of p300 in transactivation assays . Accordingly , p300 exhibits a circadian time-dependent association with Q99743 in the vasculature , which precedes peak expression of target genes . In addition , a rhythm in core histone H3 acetylation on the mPer1 promoter in vivo correlates with the cyclical expression of their mRNAs . Temporal coactivator recruitment and O60235 -dependent chromatin remodeling on the promoter of clock controlled genes in the vasculature permits the mammalian clock to orchestrate circadian gene expression . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . P12004 D1b represses breast cancer cell growth by antagonizing the action of cyclin D1a on estrogen receptor alpha-mediated transcription . Alternate splicing of the cyclin D1 gene gives rise to transcripts a and b which encode two protein isoforms cyclin D1a and cyclin D1b . P12004 D1a can substitute for estrogen to activate estrogen receptor alpha- ( ERalpha ) mediated transcription and can induce the proliferation of estrogen responsive tissues . However , little is known about the biological role of cyclin D1b in transcriptional regulation . In this study , we determined that cyclin D1b is incapable of inducing ERalpha-mediated transcription because it fails to recruit steroid receptor coactivator-1 ( Q15788 ) to ERalpha . Moreover , cyclin D1b antagonizes cyclin D1a-induced ERalpha-mediated transcription by competing with cyclin D1a for ERalpha binding . Cell proliferation assay showed that cyclin D1b repressed the ERalpha-positive breast cancer T47D cell growth . Our findings suggest that the cyclin D1b represses breast cancer cell growth by antagonizing the action of cyclin D1a on ERalpha-mediated transcription . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Farnesoid X receptor agonist for the treatment of liver and metabolic disorders : focus on 6-ethyl- DB06777 . 6-ethyl-chedeoxycholic acid ( 6E- DB06777 ) is a farnesoid X receptor ( Q96RI1 ) ligand endowed with agonistic activity under development for treatment of cholestatic liver diseases including primary biliary cirrhosis ( P10515 ) and liver-related metabolic disorders including non-alcoholic fatty liver disease ( NAFLD ) and non-alcoholic steatohepatitis ( NASH ) . Q96RI1 is a bile sensor that acts in coordination with other nuclear receptors to regulate essential steps of bile acid uptake , metabolism and excretion . 6E- DB06777 has been investigated in preclinical models of cholestasis , liver fibrosis and diet-induced atherosclerosis . In a phase II clinical trial in patients with P10515 , 6E- DB06777 met the primary endpoint of a reduction in alkaline phosphatase levels but safety data indicated that the drug exacerbated pruritus , one of the main symptoms of P10515 , suggesting that 6E- DB06777 or Q96RI1 are mediators of pruritus in humans . Treatment of patients with diabetes and liver steatosis resulted in amelioration of insulin sensitivity despite a reduction a slight reduction in HDL and increased levels of LDL were observed . These side effects on bile acids and lipid metabolism were all predicted by pre-clinical studies , suggesting that potent Q96RI1 ligands hold promise but potential side effects might limit their development .