interacts_with DB00428?
Regulation of lipogenesis by cyclin-dependent kinase 8-mediated control of P36956 . Altered lipid metabolism underlies several major human diseases , including obesity and type 2 diabetes . However , lipid metabolism pathophysiology remains poorly understood at the molecular level . P01308 is the primary stimulator of hepatic lipogenesis through activation of the SREBP-1c transcription factor . Here we identified cyclin-dependent kinase 8 ( P49336 ) and its regulatory partner cyclin C ( CycC ) as negative regulators of the lipogenic pathway in Drosophila , mammalian hepatocytes , and mouse liver . The inhibitory effect of P49336 and CycC on de novo lipogenesis was mediated through P49336 phosphorylation of nuclear SREBP-1c at a conserved threonine residue . Phosphorylation by P49336 enhanced SREBP-1c ubiquitination and protein degradation . Importantly , consistent with the physiologic regulation of lipid biosynthesis , P49336 and CycC proteins were rapidly downregulated by feeding and insulin , resulting in decreased SREBP-1c phosphorylation . Moreover , overexpression of CycC efficiently suppressed insulin and feeding-induced lipogenic gene expression . Taken together , these results demonstrate that P49336 and CycC function as evolutionarily conserved components of the insulin signaling pathway in regulating lipid homeostasis . DB00644 agonists reduce the migratory and the invasive behavior of androgen-independent prostate cancer cells by interfering with the activity of P05019 . Androgen-independent prostate carcinoma is characterized by a high proliferation rate and by a strong metastatic behavior . We have previously shown that DB00644 agonists exert a direct and specific inhibitory action on the proliferation of androgen-independent prostate cancer cells ( DU 145 ) . These compounds mainly act by interfering with the mitogenic activity of growth factors , such as the insulin-like growth factor-I ( P05019 ) . The present experiments were performed to clarify whether DB00644 agonists might also affect the migratory and the invasive behavior of androgen-independent prostate cancer cells and to define their mechanism of action . First we showed that the DB00644 agonist DB00007 reduces the migration of DU 145 cells towards a chemoattractant and their ability to invade a reconstituted basement membrane . Experiments were then performed to clarify whether the DB00644 agonist might act by interfering with the pro-metastatic activity of P05019 . We found that , in androgen-independent prostate cancer cells , DB00007 : a ) interferes with the P05019 system ( receptor protein expression and tyrosine-phosphorylation ) ; b ) abrogates the P05019 -induced phosphorylation of Akt ( a kinase previously shown by us to mediate the pro-metastatic activity of P05019 in prostate cancer cells ) ; c ) counteracts the migration and the invasive activity of the cells stimulated by P05019 ; d ) abolishes the effects of P05019 on cell morphology , on actin cytoskeleton organization and on alphavbeta3 integrin expression/cellular localization . These data indicate that DB00644 agonists , in addition to their well known antiproliferative effect , can also exert a significant inhibitory activity on the migratory and invasive behavior of androgen-independent prostate cancer cells , expressing the P30968 . DB00644 agonists act by interfering with the pro-metastatic activity of the growth factor P05019 . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . Unmet needs in ovarian cancer : dividing histologic subtypes to exploit novel targets and pathways . Ovarian cancer ( OC ) carries a poor prognosis ; however , accumulating molecular data for the major histologic subtypes may lead to subtype-specific treatment paradigms . The present review discusses what is currently understood about the major molecular and histologic subgroups of OC . Areas specifically addressed include hormonal pathways , tumor protein p53 ( P04637 ) and AT rich interactive domain 1A ( SWI-like ; O14497 ) mutation , and the breast cancer 1/2 , early onset ( P38398 /2 ) mutation/poly ( ADP-ribose ) polymerase 1 ( P09874 ) , phosphatidylinositol-4,5-bisphosphate 3- kinase , catalytic subunit alpha ( PI3KCA ) /v-akt murine thymoma viral oncogene homolog 1 ( P31749 ) /mechanistic target of rapamycin ( P42345 ) , and mitogen-activated protein kinase kinase 1 and 2 ( Q02750 /2 ) pathways . This molecular characterization only very recently has impacted clinical research efforts to develop targeted therapies for both common and rare OC subtypes . This targeted strategy is illustrated by ongoing low-grade serous , clear-cell , and mucinous subtypeexclusive clinical trials evaluating agents based on common molecular abnormalities among patients ( i.e. , P09874 inhibitors for P38398 /2 mutation-positive OC ) . This report also reviews the published clinical trial efficacy data for investigational therapies within specific subgroups , and summarizes the currently active clinical trials evaluating these agents ( e.g. , temsirolimus , sunitinib , P04637 immunotherapy , olaparib , iniparib , veliparib ) . Available data suggest that histologic profiles and molecular tumor markers are valuable resources for identifying patients who may benefit from these specific agents , and future research should focus on targeting molecules and signaling pathways that are most commonly altered in each subtype . Antihyperglycemic Activity of Houttuynia cordata Thunb. in DB00428 -Induced Diabetic Rats . Present study is an attempt to investigate plausible mechanism involved behind antidiabetic activity of standardized Houttuynia cordata Thunb. extract in streptozotocin-induced diabetic rats . The plant is used as a medicinal salad for lowering blood sugar level in North-Eastern parts of India . Oral administration of extract at 200 and 400 mg/kg dose level daily for 21 days showed a significant ( P < 0.05 ) decrease in fasting plasma glucose and also elevated insulin level in streptozotocin-induced diabetic rats . It also significantly reversed all the alterations in biochemical parameters , that is , total lipid profile , blood urea , creatinine , protein , and antioxidant enzymes in liver , pancreas , and adipose tissue of diabetic rats . Furthermore , we have demonstrated that the extract significantly reversed the expression patterns of various glucose homeostatic enzyme genes like P11168 , P14672 , and caspase-3 levels but did not show any significant effect on Q07869 - γ protein expressions . Additionally , the extract positively regulated mitochondrial membrane potential and succinate dehydrogenase ( SDH ) activity in diabetic rats . The findings justified the antidiabetic effect of H. cordata which is attributed to an upregulation of P14672 and potential antioxidant activity , which may play beneficial role in resolving complication associated with diabetes . DB09341 transporter-2 ( P11168 ) promoter mediated transgenic insulin production reduces hyperglycemia in diabetic mice . P01308 production afforded by hepatic gene therapy ( HGT ) retains promise as a potential treatment for type 1 diabetes , but successful approaches have been limited . We employed a novel and previously untested promoter for this purpose , glucose transporter-2 ( P11168 ) to drive insulin production via delivery by recombinant adeno-associated virus ( rAAV ) . In vitro , the P11168 promoter was capable of robust glucose-responsive expression in transduced HepG2 human hepatoma cells . Therefore , rAAV constructs were designed to express the furin-cleavable human preproinsulin B10 gene , under the control of the murine P11168 promoter and packaged for delivery with rAAV expressing the type 5 capsid . DB00428 -induced diabetic mice were subjected to hepatic portal vein injection immediately followed by implantation of a sustained-release insulin pellet to allow time for transgenic expression . All mice injected with the rAAV5- P11168 -fHPIB10 virus remained euglycemic for up to 35 days post-injection , with 50 % euglycemic after 77 days post-injection . In contrast , mock-injected mice became hyperglycemic within 15 days post-injection following dissolution of the insulin pellet . Serum levels of both human insulin and C-peptide further confirmed successful transgenic delivery by the rAAV5- P11168 -fHPIB10 virus . These findings indicate that the P11168 promoter may be a potential candidate for regulating transgenic insulin production for hepatic insulin gene therapy in the treatment of type I diabetes . Potential effects of peroxisome proliferator-activated receptor activator on LPS-induced lung injury in rats . Multiple factors contribute to the pathogenesis and prognosis of chronic obstructive pulmonary disease( P48444 ) , still requiring new therapeutic strategies and medications for the disease . The aim of the present study is to investigate the model of lipopolysaccharide ( LPS ) -induced chronic lung injury and hyperinflation and test therapeutic effects of peroxisome proliferator-activated receptor ( Q07869 ) -gamma agonist . Wister rats were challenged with intra-tracheal instillation of LPS at concentrations of 0.006 , 0.060 , 0.600 , and 6.000 mg/ml per kg , twice a week , for 1 , 2 , 4 and 6 weeks . Q07869 activator , 15-deoxy-Delta12,14-prostaglandin J2 ( 15D-PGJ2 ) , or vehicle ( PBS ) was administered orally and daily at the dose of 1 and 10 mg/ml per kg in animals challenged with LPS or PBS at the dose of 0.060 mg/ml per kg body weight twice a week for 4 weeks . We found that intra-tracheal exposure of LPS resulted in a dose-dependent pattern of chronic lung hyperinflation and hypertrophy , increased alveolar enlargement , reduced vascular endothelial growth factor ( P15692 ) and elevated tissue inhibitor of metalloproteinases ( P01033 ) -1 levels in bronchoalveolar lavage ( BAL ) fluid , and early changes of leukocyte influx and interferon ( IFN ) -gamma levels in bronchoalveolar lavage ( BAL ) fluid . P37231 agonist ameliorated these changes related with the dose used.LPS-induced lung disease model shows some similarities with human disease , and P37231 agonist maybe an alternative for P48444 therapy . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Differential effects of placental restriction on P01344 , Q01718 and steroidogenic enzyme mRNA levels in the foetal sheep adrenal . We have investigated the effects of restriction of placental growth on foetal adrenal growth and adrenal expression of mRNAs for P01308 -like Growth Factor II ( P01344 ) , the IGF binding protein P18065 , Steroidogenic Factor 1 ( Q13285 ) and adrenocorticotrophic hormone ( DB01285 ) receptor ( Q01718 ) and the steroidogenic cytochrome P-450 enzymes : cholesterol side chain cleavage ( P05108 ) , 17alpha-hydroxylase ( P05093 ) and 21-hydroxylase ( CYP21A1 ) ; and 3beta-hydroxysteroid dehydrogenase/Delta5Delta4 isomerase ( 3betaHSD ) . Endometrial caruncles were removed from non-pregnant ewes before mating ( placental restriction group ; PR ) . The total adrenal : foetal weight ratio was higher in PR ( n=6 foetuses ) than in control foetuses ( n=6 foetuses ) . There was no difference in plasma DB01285 concentrations between the PR and control foetuses between 130 and 140 days gestation . Adrenal P01344 mRNA levels were lower ( P < 0.05 ) in the PR group , however , adrenal P18065 mRNA levels were not different between the PR and control groups . Adrenal Q01718 mRNA levels were also lower whilst P05108 mRNA levels were increased ( P < 0.005 ) in the PR group . We conclude that foetal adrenal growth and steroidogenesis are stimulated as a consequence of foetal growth restriction and that factors other than DB01285 are important in foetal adrenal activation during chronic , sustained hypoxaemia . Mass spectrometry and hydrogen/deuterium exchange measurements of alcohol-induced structural changes in cellular retinol-binding protein type I . To bind and release its ligand , cellular retinol-binding protein type I ( P09455 ) needs to undergo conformational and dynamic changes to connect the inner , solvent-shielded cavity , where retinol is found to bind , and the outside medium . DB00162 dissociation in vitro is favoured by water/alcohol mixtures whose moderately low dielectric constants mimic a property characteristic of the membrane microenvironment where this process occurs in vivo . Apo- and holo- P09455 , in either water/methanol or water/trifluoroethanol ( TFE ) mixtures , were analyzed at equilibrium by electrospray ionization with orthogonal quadrupole time-of-flight mass spectrometry ( P19957 -Q-TOFMS ) to identify the alcohol-induced species . The questions were asked whether the presence of alcohols affects protein dynamics , as reflected by hydrogen/deuterium ( H/D ) exchange monitored by continuous-labelling experiments , and to which extent retinol dissociation influences the process . With increasing methanol , at pH near neutrality , apo- P09455 exhibits a progressively more compact conformation , resulting in reduced H/D exchange with respect to the native protein in water . DB00162 dissociation from the holo-protein did not promote hydrogen replacement . Similarly , in the presence of the low TFE concentration sufficient to cause retinol dissociation , the hydrogen exchange of the resulting apo-protein was not exalted . However , in contrast with the alkanol , higher TFE concentrations induced a transition of apo- P09455 to a new alpha-helix conformation capable of exchanging all available hydrogen atoms . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . Regulation of Q01094 -induced apoptosis by poly(ADP-ribosyl)ation . The transcription factor adenovirus E2 promoter-binding factor ( E2F ) -1 normally enhances cell-cycle progression , but it also induces apoptosis under certain conditions , including DNA damage and serum deprivation . Although DNA damage facilitates the phosphorylation and stabilization of Q01094 to trigger apoptosis , how serum starvation renders cells vulnerable to Q01094 -induced apoptosis remains unclear . Because poly(ADP-ribose) polymerase 1 ( P09874 ) , a nuclear enzyme essential for genomic stability and chromatin remodeling , interacts directly with Q01094 , we investigated the effects of P09874 on Q01094 -mediated functions in the presence and absence of serum . P09874 attenuation , which increased Q01094 transactivation , induced G2/M cell-cycle arrest under normal growth conditions , but enhanced Q01094 -induced apoptosis in serum-starved cells . Interestingly , basal P09874 activity was sufficient to modify Q01094 by poly(ADP-ribosyl)ation , which stabilized the interaction between Q01094 and the O00499 tumor suppressor in the nucleus . Accordingly , O00499 acted as an P06400 -independent Q01094 corepressor . Because Q01094 directly activates the O00499 gene promoter , O00499 curbed Q01094 activity through a negative-feedback mechanism . Conversely , when the O00499 - Q01094 interaction was abolished by P09874 suppression , Q01094 continuously increased O00499 levels . This is functionally germane , as P09874 -depletion-associated G2/M arrest was reversed by the transfection of O00499 siRNA . Moreover , PARP-inhibitor-associated anti-transformation activity was compromised by the coexpression of dominant-negative O00499 . Because serum starvation massively reduced the Q01094 poly(ADP-ribosyl)ation , we conclude that the release of O00499 from hypo-poly(ADP-ribosyl)ated Q01094 is a mechanism by which serum starvation promotes Q01094 -induced apoptosis . [ The interconnections of molecular mechanisms of hormone actions and their role in pathogenesis of obesity , insulin resistance , and diabetes mellitus ] . The various hormones , proteins and other compounds related to developing obesity , insulin resistance and type 2 diabetes are analyzed in the paper . 1 ) Leptin , ciliary neurutrophic factor , adiponectin , glucagon-like peptide 1 , peptide YY , neuromedin S , as well as the protein receptors of these hormones decrease the food consumption , increase the energy turnover , and prevent obesity , insulin resistance , and type 2 diabetes development . The mediators of these hormone and receptor actions are melanocyte stimulating hormone ( MSH ) , corticotropin-releasing hormone ( P06850 ) , and the others . 2 ) Ghrelin , endogenose cannabinoides , galanin-like peptide and the mediators of their actions : neuropeptide Y ( P01303 ) and Agouti gene related protein ( O00253 ) increase the appetite and food consumption . Peroxisome proliferation-activated receptor ( Q07869 ) performs the similar action on food intake . The activation of the first group compound functioning decreases the obesity , increases the energy turnover , facilitates the insulin action and prevents the insulin resistance and type 2 diabetes . Increasing the activities of the second group , as well as , decreasing the actions of the first one of substances induce the opposite effects and facilitate obesity , insulin resistance , and type 2 diabetes developments . The interconnections of the molecular mechanisms of so many hormone actions make the very complicated tusk to study the various endocrine disorders including diabetes mellitus as well . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . A set of consensus mammalian mediator subunits identified by multidimensional protein identification technology . The Mediator is a multiprotein transcriptional coactivator that is expressed ubiquitously in eukaryotes from yeast to mammals and is required for induction of RNA polymerase II ( pol II ) transcription by DNA binding transcription factors . In the work described here , we exploit multidimensional protein identification technology ( MudPIT ) to carry out a proteomic analysis of the subunit composition of the mammalian Mediator complex . By comparing MudPIT data sets obtained from six independent Mediator preparations immunoaffinity purified through their Nut2 ( Q9BTT4 ) , Med25 ( Q9NWA0 ) , Intersex ( Q9NX70 ) , A0JLT2 ( A0JLT2 ) , AK007855 ( Q9H204 ) , or CRSP70 ( O95402 ) subunits , we identify a set of consensus mammalian Mediator subunits . In addition , we identify as Mediator-associated proteins the P49336 -like cyclin-dependent kinase CDK11 and the Q9UHV7 -like Q71F56 protein ( Q71F56 ) , which is mutated in patients with the congenital heart defect transposition of the great arteries ( TGA ) . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Genetics of type 2 diabetes mellitus and other specific types of diabetes ; its role in treatment modalities . Type 2 diabetes mellitus ( T2DM ) is among the most challenging health issues of the 21st century and is associated with an alarming rise in the incidence . The pathophysiological processes that lead to development of T2DM are still unclear , however impairment in insulin secretion and/or action is clearly indicated . Type 2 diabetes is a polygenic disorder with multiple genes located on different chromosomes contributing to its susceptibility . Analysis of the genetic factors is further complicated by the fact that numerous environmental factors interact with genes to produce the disorder . Only a minority of cases of type 2 diabetes are caused by single gene defects and one example is maturity onset diabetes of the young ( MODY ) . Previous studies indicated that variants in genes encoding the pancreatic β-cell K+ DB00171 channel subunits Kir6.2 ( Q14654 ) and Q09428 ( Q09428 ) are associated with neonatal diabetes . Six different types of maturity onset diabetes of young ( MODY ) have been identified based on characteristic gene defect . The common Pro12Ala polymorphism in peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) gene was confirmed in several studies to be associated with type 2 diabetes as well . More recently , studies reported variants within a novel gene , Q9NQB0 , as a putative susceptibility gene for type 2 diabetes across many ethnic backgrounds around the world . MODY patients respond better to sulphonylureas and metformin , while neonatal diabetes patients with genetic mutations can be changed from insulin to oral drugs . We hereby provide a comprehensive review on the role of genetics in type 2 diabetes mellitus . P01308 -induced Q86X55 upregulation facilitates hepatocyte proliferation . Previously , we reported that Q86X55 undergoes ubiquitination-dependent degradation in renal podocytes . It was also reported that Q86X55 is necessary for fasting-induced hepatic gluconeogenesis . Based on these reports , we hypothesized that treatment with insulin , a hormone typically present under the ' fed ' condition , would inhibit gluconeogenesis via Q86X55 degradation . HepG2 cells , AML-12 cells , and rat primary hepatocytes were treated with insulin to confirm Q86X55 downregulation . Surprisingly , insulin treatment increased Q86X55 expression in all cell types examined . Furthermore , treatment with insulin increased histone 3 methylation at arginine 17 and 26 in HepG2 cells . To elucidate the role of insulin-induced Q86X55 upregulation , the HA- Q86X55 plasmid was transfected into HepG2 cells . Q86X55 overexpression did not increase the expression of lipogenic proteins generally increased by insulin signaling . Moreover , Q86X55 knockdown did not influence insulin sensitivity . P01308 is known to facilitate hepatic proliferation . Like insulin , Q86X55 overexpression increased P24941 and P11802 expression . In addition , Q86X55 knockdown reduced the number of insulin-induced G2/M phase cells . Moreover , GFP- Q86X55 overexpression increased the number of G2/M phase cells . Based on these results , we concluded that insulin-induced Q86X55 upregulation facilitates hepatocyte proliferation . These observations indicate that Q86X55 plays an important role in liver pathophysiology . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . P01308 -like growth factor binding proteins in the bovine anterior pituitary . P01308 -like growth factor binding proteins ( IGFBPs ) were characterized in bovine anterior pituitary tissue , pituitary conditioned media , and serum collected during the preovulatory and early luteal phases of the estrous cycle . Effects of in vitro treatments of pituitaries with luteinizing hormone-releasing hormone ( P01148 ) , estradiol , and progesterone on IGFBP secretion were also evaluated . Predominant IGFBPs detected in anterior pituitary tissue by immunoprecipitation , ligand blotting , and Northern blotting were P24593 ( 29 kDa ) , P18065 ( 32 kDa ) , and P17936 ( 36 and 39 kDa doublet ) . Conditioned culture media contained P24593 , a slightly larger form of P18065 ( 33 kDa ) , the 36- and 39-kDa forms of P17936 , and a more extensively glycosylated form of P17936 ( 44 kDa ) . In serum , P24593 was not readily detected , and P17936 ( 40- and 44-kDa doublet ) and P18065 ( 34 kDa ) were larger than in pituitary tissue . Levels of P18065 , -3 , and -5 in pituitary tissue decreased during the preovulatory period and were lowest in the early luteal phase . Treatment with P01148 increased P18065 levels in media twofold . Estradiol or progesterone did not alter IGFBP secretion in vitro . Predominant IGFBPs produced and released by anterior pituitary tissue were P18065 , -3 and -5 . The activity of IGFBPs fluctuates in the pituitary in association with changes in stage of estrous cycle , implicating IGFBPs as potential regulators of gonadotrope function . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . Effects of vitamin A deficiency and repletion on rat glucagon secretion . To determine whether vitamin A is involved in pancreatic alpha cell function , we tested for ( a ) effects of vitamin A deficiency on glucagon release from perifused islets and perfused pancreases , and ( b ) the presence of cytosolic retinol-binding proteins ( P09455 ) and retinoic acid-binding proteins ( CRABP ) , in the glucagon-secreting alpha cell line , ln- Q96GN5 -G9 . DB00125 19 mM plus glucose 2.8 mM-stimulated glucagon secretion was markedly impaired in islets and pancreases of vitamin A-deficient rats or rats that had at some time been cycled through vitamin A deficiency ( ever A-def ) despite repletion with retinoids for 2-4 weeks . P01308 secretion was impaired likewise . Repletion starting early in the development of vitamin A deficiency and for a longer period of time ( 18 or 60 days ) did not restore glucagon secretion , but did normalize insulin secretion . P09455 and CRABP were present in ln- Q96GN5 -G9 cells . We conclude that ( a ) vitamin A deficiency is associated with a defect in glucagon secretion ; ( b ) The defect in secretion occurs early in the course of vitamin A deficiency ; ( c ) The defect persists despite repletion ; and ( d ) The requirement of vitamin A for secretion and the presence of P09455 and CRABP in glucagon-secreting cells support a physiologic role for vitamin A at the alpha cell level . DB00428 diabetes and the expression of P11166 at the brush border and basolateral membranes of intestinal enterocytes . Changes in membrane expression of sodium-dependent glucose transporter ( P13866 ) and glucose transporter isoform ( P11168 ) protein have been implicated in the increased intestinal glucose transport in streptozotocin-diabetes . The possible involvement of P11166 in the transport response , however , has not previously been studied . Using confocal microscopy on tissue sections and Western blotting of purified brush border membrane ( BBM ) and basolateral membrane ( BLM ) , we have examined enterocyte expression of P11166 in untreated and in 1 and 21 day streptozotocin diabetic rats . In control enterocytes , P11166 was absent at the BBM and detected at low levels at the BLM . Diabetes resulted in a 4- to 5-fold increased expression of P11166 at the BLM and the protein could also be readily detected at the BBM . P01308 treatment of diabetic rats increased P11166 level at the BBM but was without effect on expression of the protein at the BLM . Central opioid inhibition of neuroendocrine stress responses in pregnancy in the rat is induced by the neurosteroid allopregnanolone . The hypothalamus-pituitary-adrenal ( Q9Y251 ) axis is the major neuroendocrine stress response system . P06850 ( P06850 ) neurons in the parvocellular paraventricular nucleus ( pPVN ) play a key role in coordinating responses of this system to stressors . The cytokine interleukin-1beta ( IL-1beta ) , mimicking infection , robustly activates these P06850 neurons via a noradrenergic input arising from the nucleus tractus solitarii ( P30990 ) . In late pregnancy , Q9Y251 axis responses to stressors , including IL-1beta , are attenuated by a central opioid mechanism that auto-inhibits noradrenaline release in the PVN . Here we show that the neuroactive progesterone metabolite allopregnanolone induces these changes in Q9Y251 responsiveness to IL-1beta in pregnancy . In late pregnancy , inhibition of 5alpha-reductase ( an allopregnanolone-synthesizing enzyme ) with finasteride restored Q9Y251 axis responses ( rapidly increased pPVN P06850 mRNA expression , DB01285 , and corticosterone secretion ) to IL-1beta . Conversely , allopregnanolone reduced Q9Y251 responses in virgin rats . In late pregnancy , activity of the allopregnanolone-synthesizing enzymes ( 5alpha-reductase and 3alpha-hydroxysteroid dehydrogenase ) was increased in the hypothalamus as was mRNA expression in the P30990 and PVN . Naloxone , an opioid antagonist , restores Q9Y251 axis responses to IL-1beta in pregnancy but had no additional effect after finasteride , indicating a causal connection between allopregnanolone and the endogenous opioid mechanism . Indeed , allopregnanolone induced opioid inhibition over Q9Y251 responses to IL-1beta in virgin rats . Furthermore , in virgin rats , allopregnanolone treatment increased , whereas in pregnant rats finasteride decreased proenkephalin-A mRNA expression in the P30990 . Thus , in pregnancy , allopregnanolone induces opioid inhibition over Q9Y251 axis responses to immune challenge . This novel opioid-mediated mechanism of allopregnanolone action may alter regulation of other brain systems in pregnancy . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Diminished phosphodiesterase-8B potentiates biphasic insulin response to glucose . DB02527 activates multiple signal pathways , crucial for the pancreatic beta-cells function and survival and is a major potentiator of insulin release . A family of phosphodiesterases ( PDEs ) terminate the DB02527 signals . We examined the expression of PDEs in rat beta-cells and their role in the regulation of insulin response . Using RT-PCR and Western blot analyses , we identified Q14432 , Q13370 , Q07343 , Q08499 , and O95263 in rat islets and in P01308 -1E cells and several possible splice variants of these PDEs . Specific depletion of Q14432 with small interfering ( si ) RNA ( siPDE3A ) led to a small ( 67 % ) increase in the insulin response to glucose in P01308 -1E cells but not rat islets . siPDE3A had no effect on the glucagon-like peptide-1 ( 10 nmol/liter ) potentiated insulin response in rat islets . Depletion in O95263 levels in rat islets using similar technology ( siPDE8B ) increased insulin response to glucose by 70 % , the potentiation being of similar magnitude during the first and second phase insulin release . The siPDE8B-potentiated insulin response was further increased by 23 % when glucagon-like peptide-1 was included during the glucose stimulus . In conclusion , O95263 is expressed in a small number of tissues unrelated to glucose or fat metabolism . We propose that O95263 , an DB07954 -insensitive DB02527 -specific phosphodiesterase , could prove a novel target for enhanced insulin response , affecting a specific pool of DB02527 involved in the control of insulin granule trafficking and exocytosis . Finally , we discuss evidence for functional compartmentation of DB02527 in pancreatic beta-cells . P10275 antagonism and an insulin sensitizer block the advancement of vaginal opening by high-fat diet in mice . Reduced hypothalamic sensitivity to steroid negative feedback may contribute to the onset of puberty . In high fat-fed rodents , the timing of vaginal opening ( VO ) is advanced , suggesting that puberty begins earlier . Because obesity can increase androgens , which interfere with normal steroid feedback in adult females , we hypothesized that androgens reduce hypothalamic sensitivity to negative feedback during puberty and that blocking androgen action would prevent advanced VO in high fat-fed mice . Age at VO was examined in mice fed high-fat or low-fat diets from weaning and treated with the androgen receptor antagonist flutamide or vehicle ( controls ) . VO was advanced in high-fat vs. low-fat controls , and flutamide blocked this advancement . VO was also delayed in low fat-fed flutamide-treated females , suggesting involvement of androgens in the timing of normal puberty . We next investigated if high-fat diet-induced insulin resistance contributes to early VO , as elevated insulin can stimulate androgen production . VO was examined in mice on either diet treated with the insulin sensitizer metformin . Metformin blocked high-fat advancement of VO but did not alter the timing of VO in low fat-fed mice . P01308 was elevated in high fat-fed females that had undergone VO compared with age-matched low fat-fed or metformin-treated animals on either diet that had not undergone VO . Together , these data suggest a model in which metabolic changes induced by high-fat diet , including transient increased circulating insulin , act in part by increasing androgen action to influence the timing of puberty in females . Di(2-ethylhexyl) phthalate induces apoptosis through peroxisome proliferators-activated receptor-gamma and P29323 1/2 activation in testis of Sprague-Dawley rats . Di(2-ethylhexyl) phthalate ( DEHP ) is a well-known hepatic and reproductive toxicant whose toxicity may be mediated by peroxisome proliferators-activated receptor ( Q07869 ) . This study examined the effects of DEHP on the expression of Q07869 -regulated genes involved in testicular cells apoptosis . Sprague-Dawley male rats were treated orally with 250 , 500 , or 750 mg/kg/d DEHP for 28 d , while control rats were given corn oil . The levels of cell cycle regulators ( P06400 , cyclins , CDKs , and P38936 ) and apoptosis-related proteins were analyzed by Western blot analysis . The role of P37231 ( P37231 ) , class B scavenger receptor type 1 ( SR-B1 ) , and P27361 /2 was further studied to examine the signaling pathway for DEHP-induced apoptosis . Results showed that the levels of pRB , cyclin D , P24941 , cyclin E , and P11802 were significantly lower in rats given 500 and 750 mg/kg/d DEHP , while levels of P38936 were significantly higher in rat testes . Dose-dependent increases in P37231 and RXRalpha proteins were observed in testes after DEHP exposure , while there was a significant decrease in RXRgamma protein levels . In addition to P37231 , DEHP also significantly increased SR-B1 mRNA and phosphorylated P27361 /2 protein levels . Furthermore , DEHP treatment induced pro-caspase-3 and cleavage of its substrate protein , poly(ADP-ribose) polymerase ( PARP ) , in a dose-dependent manner . Data suggest that DEHP exposure may induce the expression of apoptosis-related genes in testes through induction of P37231 and activation of the P27361 /2 pathway . O75791 plays an important role in glucolipotoxicity-induced apoptosis in P01308 -1 cells . OBJECTIVES : The mechanism underlying the regulation of glucolipotoxicity-induced apoptosis by MAPKs was examined in P01308 -1 cells . METHODS : The rat insulinoma cell line P01308 -1 was cotreated with glucose ( 30 mM ) and palmitic acid ( 0.2 mM ) ( GLU+PA ) . Apoptosis was assessed by cell morphology and detection of PARP cleavage . The activation of MAPKs was examined by Western blotting using specific antibodies against the phosphorylated forms of JNK , P27361 /2 , and O75791 . RESULTS : ( 1 ) Live cell imaging studies showed that treatment with GLU+PA for 72 h induced significant cell death , concomitant with P09874 cleavage and caspase-3 activation , which peaked at 96 h of treatment . ( 2 ) Western blot analysis of the activation of MAPKs during GLU+PA-induced P01308 -1 cell apoptosis showed that phosphorylation of O75791 increased gradually and reached a peak at 96 h , which coincided with P09874 cleavage . A transient increase of P29323 activation was followed by a rapid decline at 96 h , whereas JNK phosphorylation status remained unchanged in response to GLU+PA . ( 3 ) Phosphorylation of insulin receptor substrate ( P41252 ) -2 at 48 h of treatment triggered its degradation , which coincided with O75791 activation . ( 4 ) Inhibition of O75791 , but not JNK or P29323 , blocked GLU+PA-induced P01308 -1 cell apoptosis . CONCLUSIONS : O75791 may be involved in the regulation of glucolipotoxicity-induced apoptosis through the phosphorylation of Q9Y4H2 . Cytotoxic effects of fascaplysin against small cell lung cancer cell lines . Fascaplysin , the natural product of a marine sponge , exhibits anticancer activity against a broad range of tumor cells , presumably through interaction with DNA , and/or as a highly selective cyclin-dependent kinase 4 ( P11802 ) inhibitor . In this study , cytotoxic activity of fascaplysin against a panel of small cell lung cancer ( SCLC ) cell lines and putative synergism with chemotherapeutics was investigated . SCLC responds to first-line chemotherapy with platinum-based drugs/etoposide , but relapses early with topotecan remaining as the single approved therapeutic agent . Fascaplysin was found to show high cytotoxicity against SCLC cells and to induce cell cycle arrest in P55008 /0 at lower and S-phase at higher concentrations , respectively . The compound generated reactive oxygen species ( ROS ) and induced apoptotic cell death in the chemoresistant NCI-H417 SCLC cell line . Furthermore , fascaplysin revealed marked synergism with the topoisomerase I-directed camptothecin and 10-hydroxy-camptothecin . The Poly(ADP-ribose)-Polymerase 1 ( P09874 ) inhibitor Q06418 204165 antagonized the cytotoxic activity of fascaplysin , pointing to the involvement of DNA repair in response to the anticancer activity of the drug . In conclusion , fascaplysin seems to be suitable for treatment of SCLC , based on high cytotoxic activity through multiple routes of action , affecting topoisomerase I , integrity of DNA and generation of ROS . 25-Hydroxycholesterol is not a ligand for the orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) . The orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) is involved in the transcriptional regulation of all the steroid hydroxylase genes , and also regulates the transcription of the genes for Müllerian Inhibitory substance ( P03971 ) , alpha subunit of glycoprotein hormone , LHbeta , oxytocin , P30968 , Q01718 , prolactin receptor , DAX-1 , and steroidogenic acute regulatory protein . Other members of the nuclear receptor gene family , including steroid hormone , thyroid hormone , retinoic acid , Q07869 , and vitamin D receptors must bind ligand to activate transcription , but Q13285 has been considered to be an orphan nuclear receptor because , when identified , it had no known ligand . A recent publication suggested that transcriptional regulation by Q13285 , expressed in a non-steroidogenic CV-1 cells , could be activated by oxysterols suggesting that these compounds could serve as natural ligands for Q13285 . We now demonstrate that 25-hydroxycholesterol , either added exogenously or synthesized endogenously in steroidogenic mouse Leydig MA-10 cells , did not act as a ligand for Q13285 , as it did not increase transcription from six different Q13285 -dependent DNA sequences . Furthermore , the abundance of these oxysterols in MA-10 cells was much less than concentrations needed for activation of Q13285 in CV-1 cells , indicating that Q13285 is not constitutively bound by ligand in MA-10 cells . Thus , in steroidogenic cells , transcriptional regulation of the steroid hydroxylase genes by Q13285 does not depend upon the presence of 25-hydroxycholesterol , and is not modified by its presence . Cloning of chick cellular retinol-binding protein , type II and comparison to that of some mammals : expression of the gene at different developmental stages , and possible involvement of RXRs and Q07869 . We cloned chick cellular retinol-binding protein , type two ( P09455 II ) cDNA and compared it with those of some mammals . The deduced amino acid sequence showed that chick P09455 II was one amino acid greater in size than those of mammals , and the nucleotide sequence of chick P09455 II shared 72 % -75 % similarity with those of mammals . RNA blot hybridization analysis showed that P09455 II transcript of 0.7 kb was first detected in the duodenum of day-18 embryonic chick , and exhibited a rapid increase during 24 hr around the hatching . Northern blot hybridization also revealed that the transcripts of two types of retinoid X receptors ( RXR alpha and RXR gamma ) and peroxisome proliferator-activated receptor ( Q07869 ) were expressed in the chick duodenum at hatching . The organ culture of day 16 embryonic chick duodenum showed that the addition of 9-cis retinoic acid in the medium caused a significant increase in P09455 II mRNA levels . In addition , arachidonic acid , from which putative ligands for Q07869 were supposed to be generated , was accumulated around hatching in the duodenum . The results may suggest that the abrupt increase of the P09455 II gene expression in the chick duodenum around hatching may be related with RXRs and/or Q07869 . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . The peroxisome proliferator-activated receptor ( Q07869 ) β/δ agonist GW501516 inhibits P05231 -induced signal transducer and activator of transcription 3 ( P40763 ) activation and insulin resistance in human liver cells . AIM/HYPOTHESIS : P05231 induces insulin resistance by activating signal transducer and activator of transcription 3 ( P40763 ) and upregulating the transcription of its target gene O14543 . Here we examined whether the peroxisome proliferator-activated receptor ( Q07869 )β/δ agonist GW501516 prevented activation of the P05231 - P40763 -suppressor of cytokine signalling 3 ( O14543 ) pathway and insulin resistance in human hepatic HepG2 cells . METHODS : Studies were conducted with human HepG2 cells and livers from mice null for Pparβ/δ ( also known as Ppard ) and wild-type mice . RESULTS : GW501516 prevented P05231 -dependent reduction in insulin-stimulated v-akt murine thymoma viral oncogene homologue 1 ( AKT ) phosphorylation and in P35568 and Q9Y4H2 protein levels . In addition , treatment with this drug abolished P05231 -induced P40763 phosphorylation of Tyr⁷⁰⁵ and Ser⁷²⁷ and prevented the increase in O14543 caused by this cytokine . Moreover , GW501516 prevented P05231 -dependent induction of extracellular-related kinase 1/2 ( P27361 /2 ) , a serine-threonine protein kinase involved in serine P40763 phosphorylation ; the livers of Pparβ/δ-null mice showed increased Tyr⁷⁰⁵- and Ser⁷²⁷- P40763 as well as phospho- P27361 /2 levels . Furthermore , drug treatment prevented the P05231 -dependent reduction in phosphorylated AMP-activated protein kinase ( AMPK ) , a kinase reported to inhibit P40763 phosphorylation on Tyr⁷⁰⁵ . In agreement with the recovery in phospho-AMPK levels observed following GW501516 treatment , this drug increased the AMP/ DB00171 ratio and decreased the DB00171 /ADP ratio . CONCLUSIONS/INTERPRETATION : Overall , our findings show that the PPARβ/δ activator GW501516 prevents P05231 -induced P40763 activation by inhibiting P27361 /2 phosphorylation and preventing the reduction in phospho-AMPK levels . These effects of GW501516 may contribute to the prevention of cytokine-induced insulin resistance in hepatic cells . Activation of gonadotropin-releasing hormone receptors induces a long-term enhancement of excitatory postsynaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus . Whole-cell patch-clamp recordings were made from P00915 pyramidal neurons of the rat hippocampus to study the modulation of gonadotropin-releasing hormone ( DB00644 ) on synaptic transmission mediated by ionotropic glutamate receptors . DB00007 ( 10(-9)-10(-7) M ) , a specific DB00644 analog , concentration-dependently elicited a long-lasting potentiation of excitatory postsynaptic currents ( EPSCs ) mediated by ionotropic glutamate receptors . P30968 -induced synaptic potentiation was blocked by 1 microM [ Acetyl-3,4-dehydro-Pro1,D-p-F-Phe2,D-Trp3,6 ] - P01148 , a specific P30968 antagonist . Furthermore , P30968 -induced synaptic potentiation was associated with the stimulation of protein kinase C ( PKC ) , being considerably attenuated by a potent PKC inhibitor ( 30 microM H-7 ) . The results suggest a long-term enhanced modulation of DB00644 on synaptic transmission mediated by ionotropic glutamate receptors , possibly via the actions of PKC in the hippocampus that is an important integrative system in the regulation of reproductive processes . Poly(ADP-ribose) polymerase-1 ( P09874 ) is required in murine cell lines for base excision repair of oxidative DNA damage in the absence of P06746 . Oxidative DNA base damage is mainly corrected by the base excision repair ( BER ) pathway , which can be divided into two subpathways depending on the length of the resynthetized patch , either one nucleotide for short patch BER or several nucleotides for long patch BER . The role of proteins in the course of BER processes has been investigated in vitro using purified enzymes and cell-free extracts . In this study , we have investigated the repair of 8-oxo-7,8-dihydroguanine ( 8-oxoG ) in vivo using wild-type , polymerase beta(-/-) ( Polbeta(-/-) ) , poly(ADP-ribose) polymerase-1(-/-) ( P09874 (-/-) ) , and Polbeta(-/-) P09874 (-/-) 3T3 cell lines . We used non replicating plasmids containing a 8-oxoG:C base pair to study the repair of the lesion located in a transcribed sequence ( TS ) or in a non-transcribed sequence ( P30990 ) . The results show that 8-oxoG repair in TS is not significantly impaired in cells deficient in Polbeta or P09874 or both . Whereas 8-oxoG repair in P30990 is normal in Polbeta-null cells , it is delayed in P09874 -null cells and greatly impaired in cells deficient in both Polbeta and P09874 . The removal of 8-oxoG and presumably the cleavage at the resulting apurinic/apyrimidinic site are not affected in the P09874 (-/-)Polbeta(-/-) cell lines . However , 8-oxoG repair is incomplete , yielding plasmid molecules with a nick at the site of the lesion . Therefore , P09874 (-/-)Polbeta(-/-) cell lines can not perform 5'-dRP removal and/or DNA repair synthesis . Furthermore , the poly(ADP-ribosyl)ation activity of P09874 is essential for 8-oxoG repair in a Polbeta(-/-) context , because expression of the catalytically inactive P09874 ( E988K ) mutant does not restore 8-oxoG repair , whereas an wild type P09874 does . Coactivators for the orphan nuclear receptor RORalpha . A mutation in the nuclear orphan receptor RORalpha results in a severe impairment of cerebellar development by unknown mechanisms . We have shown previously that RORalpha contains a strong constitutive activation domain in its C terminus . We therefore searched for mammalian RORalpha coactivators using the minimal activation domain as bait in a two-hybrid screen . Several known and putative coactivators were isolated , including glucocorticoid receptor-interacting protein-1 ( Q9Y3R0 ) and peroxisome proliferator-activated receptor ( Q07869 ) -binding protein ( PBP/ Q15648 / Q15648 ) . These interactions were confirmed in vitro and require the intact activation domain of RORalpha although different requirements for interaction with Q9Y3R0 and PBP were detected . Even in the absence of exogenous ligand , RORalpha interacts with a complex or complexes of endogenous proteins , similar to those that bind to ligand-occupied thyroid hormone and vitamin D receptors . Both PBP and Q9Y3R0 were shown to be present in these complexes . Thus we have identified several potential RORalpha coactivators that , in contrast to the interactions with hormone receptors , interact with RORalpha in yeast , in bacterial extracts , and in mammalian cells in vivo and in vitro in the absence of exogenous ligand . Q9Y3R0 functioned as a coactivator for the RORalpha both in yeast and in mammalian cells . Thus , Q9Y3R0 is the first proven coactivator for RORalpha . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . DB00428 -nicotinamide-induced diabetes in the rat . Characteristics of the experimental model . Administration of both streptozotocin ( Q11206 ) and nicotinamide ( NA ) has been proposed to induce experimental diabetes in the rat . Q11206 is well known to cause pancreatic B-cell damage , whereas NA is administered to rats to partially protect insulin-secreting cells against Q11206 . Q11206 is transported into B-cells via the glucose transporter P11168 and causes DNA damage leading to increased activity of poly(ADP-ribose) polymerase ( P09874 ) to repair DNA . However , exaggerated activity of this enzyme results in depletion of intracellular NAD(+) and DB00171 , and the insulin-secreting cells undergo necrosis . The protective action of NA is due to the inhibition of P09874 activity . NA inhibits this enzyme , preventing depletion of NAD(+) and DB00171 in cells exposed to Q11206 . Moreover , NA serves as a precursor of NAD(+) and thereby additionally increases intracellular NAD(+) levels . The severity of diabetes in experimental rats strongly depends on the doses of Q11206 and NA given to these animals . Therefore , in diabetic rats , blood glucose may be changed in a broad range -- from slight hyperglycemia to substantial hyperglycemia compared with control animals . Similarly , blood insulin may be only slightly decreased or substantial hypoinsulinemia may be induced . In vitro studies demonstrated that the insulin-secretory response to glucose is attenuated in Q11206 -NA-induced diabetic rats compared with control animals . This is due to reduced B-cell mass as well as metabolic defects in the insulin-secreting cells . Results of numerous experiments have demonstrated that this model of diabetes is useful in studies of different aspects of diabetes . P01308 reverses growth hormone-induced homologous desensitization . Growth hormone ( GH ) is secreted in a pulsatile pattern to promote body growth and metabolism . GH exerts its function by activating several signaling pathways , including O60674 / P35610 and MEK/ P29323 . P27361 /2 activation by GH plays important roles in gene expression , cell proliferation , and growth . We previously reported that in rat H4IIE hepatoma cells after an initial GH exposure , a second GH exposure induces P42229 phosphorylation but not P27361 /2 phosphorylation ( Ji , S. , Frank , S. J. , and Messina , J. L. ( 2002 ) J. Biol. Chem. 277 , 28384-28393 ) . In this study the mechanisms underlying GH-induced homologous desensitization were investigated . A second GH exposure activated the signaling intermediates upstream of MEK/ P29323 , including O60674 , Ras , and P04049 . This correlated with recovery of P10912 levels , but was insufficient for GH-induced phosphorylation of Q02750 /2 and P27361 /2 . P01308 restored the ability of a second GH exposure to induce phosphorylation of Q02750 /2 and P27361 /2 without altering P10912 levels or GH-induced phosphorylation/activation of O60674 and P04049 . GH and insulin synergized in promoting cell proliferation . Further investigation suggested that insulin increased the amount of MEK bound to Q8IVT5 ( kinase suppressor of Ras ) and restored GH-induced tyrosine phosphorylation of Q8IVT5 . Previous GH exposure also induced desensitization of P42224 and P40763 phosphorylation , but this desensitization was not reversed by insulin . Thus , insulin-regulated resensitization of GH signaling may be necessary to reset the complete response to GH after a normal , physiologic pulse of GH . Tetramethoxychalcone , a chalcone derivative , suppresses proliferation , blocks cell cycle progression , and induces apoptosis of human ovarian cancer cells . In the present study , we investigated the in vitro antitumor functions of a synthetic chalcone derivative 4,3',4',5'- tetramethoxychalcone ( TMOC ) in ovarian cancer cells . We found that TMOC inhibited the proliferation and colony formation of cisplatin sensitive cell line A2780 and resistant cell line A2780/ DB00515 , as well as ovarian cancer cell line SKOV3 in a time- and dose-dependent manner . Treatment of A2780 cells with TMOC resulted in G0/ P55008 cell cycle arrest through the down-regulation of cyclin D1 and P11802 , and the up-regulation of p16 , P38936 and p27 proteins . We demonstrated that TMOC might induce cell apoptosis through suppressing Bcl-2 and Bcl-xL , but enhancing the expression of Bax and the cleavage of P09874 . Treatment of TMOC also reduced the invasion and migration of A2780 cells . Finally , we found that TMOC inhibited the constitutive activation of P40763 signaling pathway and induced the expression of the tumor suppressor P60484 regardless of the p53 status in cell lines . These data suggest that TMOC may be developed as a potential chemotherapeutic agent to effectively treat certain cancers including ovarian cancer . P09874 inhibition-induced activation of P19957 -kinase-Akt pathway promotes resistance to taxol . PARP inhibitors combined with DNA-damage inducing cytostatic agents can lead to effective tumor therapy . However , inhibition of poly(ADP-ribose) polymerase ( P09874 ; EC 2.4.2.30 ) induces the activation of P19957 -kinase-Akt pathway , which can counteract the effectiveness of this therapy . To understand the role of Akt activation in the combined use of cytostatic agent and PARP inhibition , we used taxol ( paclitaxel ) as an antineoplastic agent , which targets microtubules and up-regulates mitochondrial ROS production , together with ( i ) pharmacological inhibition ( PJ-34 ) , ( ii ) siRNA knock-down and ( iii ) transdominant expression of the DNA binding domain of P09874 . In all cases , P09874 inhibition leads to suppressed poly-ADP-ribosylation of nuclear proteins , prevention of NAD(+) depletion and significant resistance against taxol induced caspase-3 activation and apoptotic cell death . Paclitaxel induced a moderate increase in Akt activation , which was significantly augmented by PARP inhibition , suggesting that PARP inhibition-induced Akt activation could be responsible for the cytostatic resistance . When activation of the P19957 -kinase-Akt pathway was prevented by LY-294002 or Akt Inhibitor IV , the cytoprotective effect of PARP inhibition was significantly diminished showing that the activation of P19957 -kinase-Akt cascade had significantly contributed to the cytostatic resistance . Our study demonstrates that drug-induced drug resistance can be responsible for the reduced efficacy of antitumor treatment . Although inhibition of P09874 can promote cell death in tumor cells by the inhibition of DNA repair , PARP-inhibition promoted activation of the P19957 -kinase-Akt pathway can counteract this facilitating effect , and can cause cytostatic resistance . We suggest augmenting PARP inhibition by the inhibition of the P19957 -kinase-Akt pathway for antitumor therapy . Systemic poly(ADP-ribose) polymerase-1 activation , chronic inflammation , and oxidative stress in P48444 patients . Oxidative stress and systemic inflammation in chronic obstructive pulmonary disease ( P48444 ) strongly suggest a role for the nuclear enzyme poly(ADP-ribose) polymerase-1 ( P09874 , E.C.2.4.2.30 ) in the disease pathophysiology . P09874 is highly activated by reactive oxygen species-induced DNA strand breaks , upon which it forms extensive poly(ADP-ribose) ( PAR ) polymers from its substrate NAD(+) . We hypothesized that in P48444 , chronic inflammation and oxidative stress would lead to systemic P09874 activation and to a reduced NAD(+) status . In a patient-control study , systemic P09874 activation was assessed by immunofluorescent detection of PAR polymers in peripheral blood lymphocytes . The percentage of PAR polymer-positive lymphocytes appeared to be higher in P48444 patients ( 27 +/- 3 % ) than in healthy age-matched controls ( 17 +/- 2 % , p < .05 ) . Trolox equivalent antioxidant capacity ( TEAC ) of deproteinized plasma ( p < .001 ) , plasma uric acid ( p < .05 ) , as well as blood NAD(+) ( p < .01 ) of stable P48444 patients were significantly reduced when compared to controls . In addition , levels of proinflammatory cytokines P05231 , P10145 , and sICAM-1 were increased ( p < .005 ) in P48444 patients . In this study , evidence was found for the presence of systemic inflammation , chronic oxidative stress , and systemic P09874 activation in stable P48444 patients . These data support a contribution of oxidative stress-induced P09874 activation to the pathophysiology of P48444 . Biological and immunological studies of bovine hypothalamic DB05394 . P06850 B ( CRF-B ) is a peptide(s) isolated from bovine hypothalamic extracts by Sephadex G-100 chromatography on the basis of its ability to stimulate secretion of adrenocorticotropin ( DB01285 ) in vitro and in vivo . It is similar in molecular size to the 41-residue ovine CRF ( oCRF ) or rat CRF ( rCRF ) recently elucidated and appears to be their bovine counterpart . Immunoreactivity of CRF-B was examined in homologous radioimmunoassays ( RIAs ) for oCRF or rCRF , using several anti-oCRF and anti-rCRF antibodies . CRF-B cross-reacted well with anti-oCRF antibodies but poorly with anti-rCRF antibodies . Purification of CRF-B with preparative isoelectric focusing yielded four CRF peaks , B-1 ( pH 4.7 ) , B-2 ( pH 5.5 ) , B-3 ( pH 6.3 ) , and B-4 ( pH 7.0 ) , which accounted for 16 , 30 , 46 , and 8 % of the total immunoreactivity , respectively . CRF B-2 , B-3 , and B-4 showed both immunological activity and biological activity in vitro ( cell culture assay ) and in vivo ( Arimura assay ) , whereas CRF B-1 showed only immunoreactivity . Their relative bioactivity/immunoreactivity ratios were 0 ( B-1 ) , 1 ( B-2 ) , 1 ( B-3 ) , and 3 ( B-4 ) . All of these CRF-B subtypes exhibited RIA displacement curves parallel to that for the oCRF standard and coeluted with oCRF on Sephadex G-100 chromatography , which suggests that their molecular modifications are relatively minor . Effect of growth factors on nuclear and mitochondrial ADP-ribosylation processes during astroglial cell development and aging in culture . Epidermal growth factor ( P01133 ) , basic fibroblast growth factor ( P09038 ) , insulin-like growth factor-I ( P05019 ) and insulin ( P01308 ) are powerful mitogens and may regulate gene expression in cultured astrocytes by ADP-ribosylation process . Nuclear poly-ADP ribose polymerase ( PARP ) and mitochondrial monoADP-ribosyltransferase ( P09874 ) are the key enzymes involved in poly-ADP-ribosylation and mono ADP-ribosylation , respectively . In this investigation the effect of P01133 , P09038 , P05019 or P01308 on nuclear PARP and mitochondrial P09874 activities were assessed in nuclei and mitochondria purified from developing ( 30 DIV ) or aging ( 90 and 190 DIV ) primary rat astrocyte cultures . A marked increase of PARP activity in P09038 or P05019 treated astroglial cell cultures at 30 DIV was found . Nuclear PARP and mitochondrial P09874 activities were greatly stimulated by treatment with P01133 or P01308 alone or together in astrocyte cultures at 30 DIV . Nuclear PARP and mitochondrial P09874 activities showed a more remarkable increase in control untreated astrocyte cultures at 190 DIV than at 90 DIV . These findings suggest that ADP-ribosylation process is involved in DNA damage and repair during cell differentiation and aging in culture . Twelve hours treatment with P01133 , P01308 or P09038 significantly stimulated nuclear PARP and mitochondrial P09874 activities in 190 DIV aging astrocyte cultures . The above results indicate that P01133 , P01308 and P09038 may play a crucial role in the post-translational modification of chromosomal proteins including ADP-ribosylation process in in vitro models . This suggests that growth factors regulate genomic stability in glial cells during development and maturation , stimulating nuclear and mitochondrial ADP-ribosylation processes in developing or aging astrocyte cultures . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway . DB00624 supplementation increases skeletal muscle mass and decreases fat mass ; however , the underlying mechanisms are unknown . We hypothesized that testosterone regulates body composition by promoting the commitment of mesenchymal pluripotent cells into myogenic lineage and inhibiting their differentiation into adipogenic lineage . Mouse C3H 10T1/2 pluripotent cells were treated with testosterone ( 0-300 nM ) or dihydrotestosterone ( DB02901 , 0-30 nM ) for 0-14 d , and myogenic conversion was evaluated by immunocytochemical staining for early ( MyoD ) and late ( myosin heavy chain II ; MHC ) myogenic markers and by measurements of MyoD and MHC mRNA and protein . Adipogenic differentiation was assessed by adipocyte counting and by measurements of peroxisomal proliferator-activated receptor gamma 2 ( Q07869 gamma 2 ) mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . The number of MyoD+ myogenic cells and MHC+ myotubes and MyoD and MHC mRNA and protein levels increased dose dependently in response to testosterone and DB02901 treatment . Both testosterone and DB02901 decreased the number of adipocytes and down-regulated the expression of Q07869 gamma 2 mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . P10275 mRNA and protein levels were low at baseline but increased after testosterone or DB02901 treatment . The effects of testosterone and DB02901 on myogenesis and adipogenesis were blocked by bicalutamide . Therefore , testosterone and DB02901 regulate lineage determination in mesenchymal pluripotent cells by promoting their commitment to the myogenic lineage and inhibiting their differentiation into the adipogenic lineage through an androgen receptor-mediated pathway . The observation that differentiation of pluripotent cells is androgen dependent provides a unifying explanation for the reciprocal effects of androgens on muscle and fat mass in men .