interacts_with DB00624?
Drosophila Answers to Q13148 Proteinopathies . Initially implicated in the pathogenesis of P13569 and HIV-1 transcription , nuclear factor Q13148 was subsequently found to be involved in the origin and development of several neurodegenerative diseases . In 2006 , in fact , it was reported for the first time the cytoplasmic accumulation of Q13148 in ubiquitin-positive inclusions of P35858 and FTLD patients , suggesting the presence of a shared underlying mechanism for these diseases . Today , different animal models of Q13148 proteinopathies are available in rodents , nematodes , fishes , and flies . Although these models recapitulate several of the pathological features found in patients , the mechanisms underpinning the progressive neuronal loss observed in Q13148 proteinopathies remain to be characterized . Compared to other models , Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies . At present , the development of Q13148 -related Drosophila models has further strengthened the hypothesis that both Q13148 " loss-of-function " and " gain-of-function " mechanisms can contribute to disease . The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies , along with the information they have generated , towards a better understanding of the mechanisms underlying Q13148 proteinopathies . Androgen receptors . DB00624 and its active metabolite dihydrotestosterone exert their influence on target cells through a specific intracellular protein receptor . Structural abnormalities of this receptor lead to a diminished androgen action within the cell and result in the syndrome of androgen insensitivity . Androgen insensitivity is classified on the basis of whether the insensitivity is complete or partial and whether the androgen receptor is normally present ( AR(+) ) , absent ( AR(-) ) or diminished ( AR(+/-) ) . All patients with androgen insensitivity have normal or high plasma levels of testosterone and elevated serum LH . Patients with complete androgen insensitivity are phenotypically female . The clinical presentation of partial androgen insensitivity is variable , ranging from a minimal amount of virilization to a completely masculine appearance . All patients described with a syndrome of androgen insensitivity are infertile . The influence of androgen receptor function in the pathogenesis of benign prostatic hypertrophy is being investigated . P10275 content is also being studied as a possible marker of responsiveness to hormonal therapy in prostatic carcinoma . Gene expression in sexually dimorphic muscles in sheep . DB00624 is known to act differentially on skeletal muscle from different regions of the body . Two genes likely to mediate the testosterone effect are insulin-like growth factor I ( P05019 ) , an important growth regulator acting in an autocrine and paracrine way , and androgen receptor ( AR ) , because receptor density could account for differential muscle growth . Another muscle-specific gene that may play a role in differential muscle growth is myostatin , a member of the transforming growth factor-beta superfamily , shown to be a negative regulator of skeletal muscle mass . The objective of this study was to quantify and compare the steady state expression of these three genes in two different skeletal muscles in sheep . Eleven Dorset rams were slaughtered after reaching puberty and total RNA was extracted from samples of semitendinosus and splenius muscles . P05019 mRNA was measured using a competitive reverse-transcription-polymerase chain reaction . P10275 and myostatin mRNA were measured by a ribonuclease protection assay ( RPA ) with standard curves . The means ( attomoles/microg RNA ) for splenius and semitendinosus muscles were 1.39 and 1.02 ( SE = 0.14 ) , 4.05 and 2.96 ( SE = 0.24 ) , and 4.30 and 3.85 ( SE = 0.37 ) for P05019 , AR , and myostatin , respectively . The difference between the two muscles was significant for P05019 and AR mRNA levels with higher levels in the splenius but not significant for myostatin . Our results show that locally produced P05019 and the regulation of AR expression may be important for sexually dimorphic muscle growth patterns . 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 . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . P10275 signalling in peritubular myoid cells is essential for normal differentiation and function of adult Leydig cells . DB00624 synthesis depends on normal Leydig cell ( LC ) development , but the mechanisms controlling this development remain unclear . We recently demonstrated that androgen receptor ( AR ) ablation from a proportion of testicular peritubular myoid cells ( PTM-ARKO ) did not affect LC number , but resulted in compensated LC failure . The current study extends these investigations , demonstrating that PTM AR signalling is important for normal development , ultrastructure and function of adult LCs . Notably , mRNAs for LC markers [ e.g. steroidogenic factor 1 ( Nr5a1 ) , insulin-like growth factor ( Igf-1 ) and insulin-like factor 3 ( Insl3 ) ] were significantly reduced in adult PTM-ARKOs , but not all LCs were similarly affected . Two LC sub-populations were identified , one apparently ' normal ' sub-population that expressed adult LC markers and steroidogenic enzymes as in controls , and another ' abnormal ' sub-population that had arrested development and only weakly expressed P51460 , luteinizing hormone receptor , and several steroidogenic enzymes . Furthermore , unlike ' normal ' LCs in PTM-ARKOs , the ' abnormal ' LCs did not involute as expected in response to exogenous testosterone . Differential function of these LC sub-populations is likely to mean that the ' normal ' LCs work harder to compensate for the ' abnormal ' LCs to maintain normal serum testosterone . These findings reveal new paracrine mechanisms underlying adult LC development , which can be further investigated using PTM-ARKOs . Axotomy transiently down-regulates androgen receptors in motoneurons of the spinal nucleus of the bulbocavernosus . DB00624 is an important trophic factor for motoneurons in the spinal nucleus of the bulbocavernosus ( SNB ) , and SNB motoneurons are more responsive to testosterone than are other motoneurons . Axonal injury during early postnatal life prevents the normal development of steroid-sensitivity by adult SNB motoneurons . Axonal injury also causes changes in the expression by motoneurons of a wide range of proteins , including the up-regulation of trophic factor receptors . We have used a polyclonal antibody ( PG-21 ; G.S. Prins ) to study the expression of androgen receptors in SNB motoneurons after axonal injury . PG-21 labeled motoneuronal nuclei in the lower lumbar spinal cord of rats in a pattern that matched autoradiographic reports of androgen accumulation in this region of the nervous system . A population of numerous , small cells located dorsal to the central canal also showed evidence of androgen receptor expression . Cutting the axons of SNB motoneurons in adulthood or in development caused a decrease in androgen receptor immunoreactivity in SNB motoneurons . This is the first report that a trophic factor receptor in motoneurons is down-regulated after axonal injury , and is interesting in light of reports that testosterone treatment can facilitate motoneuronal regeneration after nerve cut . P10275 levels subsequently returned to normal , regardless of the age at axotomy , providing no evidence for a lasting effect of developmental axotomy on androgen receptor levels in SNB motoneurons . Thus , axotomy-induced down-regulation of androgen receptors does not underlie the inability of SNB motoneurons to respond to androgen treatment several months after pudendal nerve cut in development . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Stromal cell-derived factor-1 ( P48061 ) enhances cells invasion by αvβ6 integrin-mediated signaling in ovarian cancer . Ovarian carcinoma is a common gynecological malignancy and a great threat to health as a result of metastasis . The chemokine stromal-derived factor ( P48061 ) plays multiple roles in tumor pathogenesis . However , the precise molecular mechanism underlying P48061 -induced ovarian cancer cell invasion is still undefined . αvβ6 integrin is an important factor in tumor progression . Therefore , we speculate that P48061 -enhanced ovarian cancer cell invasion is related to αvβ6 integrin-mediated signaling . After culturing with P48061 , an obvious time- and dose-dependent increase in αvβ6 integrin was demonstrated . Furthermore , CXC receptor 4 ( P61073 ) was responsible for P48061 -induced αvβ6 integrin expression . Simultaneously , P48061 was found to dramatically enhance extracellular matrix degradation via urokinase-type plasminogen activator ( uPA ) expression and cell invasion by αvβ6 integrin expression ; these reinforce failed to be increased when pretreatment was performed with the P61073 inhibitor DB06809 or anti-αvβ6 integrin antibody , respectively . In addition , αvβ6 integrin induced the phosphorylation of p38 MAPK and P19957 K/Akt , contributing to the up-regulation of uPA , as treatment with the specific inhibitor for p38 mitogen-activated protein kinases ( MAPK ) ( SB203580 ) or phosphatidylinositol 3-kinase ( P19957 K ) /Akt ( LY294002 ) strikingly abrogated uPA expression . Taken together , these results demonstrated that P48061 enhanced ovarian cancer cell invasion through αvβ6 integrin-mediated uPA expression via the p38 MAPK and P19957 K/Akt pathway . Consequently , our findings will provide a new explanation about how P48061 aggravates the pathogenesis of ovarian cancer . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . DB00624 potentiates the hypoxic ventilatory response of adult male rats subjected to neonatal stress . Neonatal stress disrupts development of homeostatic systems . During adulthood , male rats subjected to neonatal maternal separation ( Q5H8A3 ) are hypertensive and show a larger hypoxic ventilatory response ( HVR ) , with greater respiratory instability during sleep . Neonatal stress also affects sex hormone secretion ; hypoxia increases circulating testosterone of Q5H8A3 ( but not control ) male rats . Given that these effects of Q5H8A3 are not observed in females , we tested the hypothesis that testosterone elevation is necessary for the stress-related increase of the HVR in adult male rats . Pups subjected to Q5H8A3 were placed in an incubator for 3 h per day from postnatal day 3 to 12 . Control pups remained undisturbed . Rats were reared until adulthood , and the HVR was measured by plethysmography ( fractional inspired O2 = 0.12 , for 20 min ) . We used gonadectomy to evaluate the effects of reducing testosterone on the HVR . Gonadectomy had no effect on the HVR of control animals but reduced that of Q5H8A3 animals below control levels . Immunohistochemistry was used to quantify androgen receptors in brainstem areas involved in the HVR . P10275 expression was generally greater in Q5H8A3 rats than in control rats ; the most significant increase was noted in the caudal region of the nucleus tractus solitarii . We conclude that the abnormal regulation of testosterone is important in stress-related augmentation of the HVR . The greater number of androgen receptors within the brainstem may explain why Q5H8A3 rats are more sensitive to testosterone withdrawal . Based on the similarities of the cardiorespiratory phenotype of Q5H8A3 rats and patients suffering from sleep-disordered breathing , these results provide new insight into its pathophysiology , especially sex-based differences in its prevalence . Genetics of idiopathic disseminated bronchiectasis . Bronchiectasis is an abnormal dilation of bronchi , consequent to the destruction of their walls . It is included in the category of obstructive pulmonary diseases , along with chronic obstructive pulmonary disease ( P48444 ) , asthma , and cystic fibrosis . In approximately 50 % of cases , bronchiectasis is associated with underlying conditions ; in the remainder , known causes are not ascertainable ( idiopathic bronchiectasis ) . A search for genetic determinants of this phenotype , with the cystic fibrosis gene as a candidate , has been performed by three independent groups . The results of this search agreed on the association of bronchiectasis with cystic fibrosis gene mutations and polymorphisms . The cystic fibrosis gene is also associated with bronchiectasis due to rheumatoid arthritis and allergic bronchopulmonary aspergillosis . A few other genes have been investigated in idiopathic bronchiectasis , with negative results . Idiopathic bronchiectasis is , therefore , to be considered as an obstructive multifactorial disorder belonging to the category of cystic fibrosis monosymptomatic diseases ( or P13569 -opathies ) , whose pathogenesis is influenced by environmental factors and other undetermined genes . DB00624 and androgen receptor in human nephrolithiasis . PURPOSE : We investigated the relationship of kidney calculi with plasma free and total testosterone , and androgen receptor up-regulation in the kidneys of men with nephrolithiasis . MATERIALS AND METHODS : Male patients with kidney stone and healthy men were included in the study . Blood was collected in a tube containing 2 % heparin in the morning . Total and free serum testosterone was measured by enzyme linked immunosorbent assay . All patients underwent percutaneous nephrostolithotomy . At the end of the procedure ultrasound guided puncture biopsy was done to acquire kidney tissue . Normal kidney tissue obtained at autopsy served as the control . P10275 was detected in kidney tissue by immunohistochemistry . Stone composition was analyzed in each patient . RESULTS : The study included 37 male patients 22 to 39 years old and 31 healthy men 24 to 37 years old . All calculi were composed of calcium oxalate monohydrate or calcium oxalate dihydrate and a few also contained protein or uric acid . Mean±SD serum total and free testosterone was 13.29±4.79 ng/ml and 63.23±28.58 pg/ml in patients , and 7.30±0.82 ng/ml and 35.59±24.91 pg/ml in healthy men , respectively ( each p < 0.001 ) . Immunohistochemistry revealed androgen receptor up-regulation in the kidneys of patients with nephrolithiasis . CONCLUSIONS : Our data suggest the important role of enhanced androgen signaling in human nephrolithiasis . Altered levels of angiopoietin 1 and tie 2 are associated with androgen-regulated vascular regression and growth in the ventral prostate in adult mice and rats . The involution of the rat ventral prostate gland after castration could be caused by primary changes in the vasculature . To explore the mechanisms , we studied the effects of castration and testosterone treatment on the vasculature in the ventral prostate in adult rats and mice . P10275 expression , vascular morphology , and the expression of angiopoietin ( ang ) 1 and 2 and their receptor tie 2 were examined 1 , 3 , and 7 d after castration and after testosterone treatment of castrated animals using stereological methods , immunohistochemistry , laser capture microdissection , and Western blotting . One day after castration , the percentage of blood vessels covered with smooth muscle actin , endothelial cell proliferation , and vascular volume had decreased , whereas endothelial cell apoptosis had increased . Simultaneously , ang 1 and tie 2 protein levels decreased . Nuclear expression of androgen receptor was observed not only in glandular and stroma smooth muscle cells but also in the mural cells of prostate arteries and veins and was markedly down-regulated already 1 d after castration . DB00624 administration of castrated mice and rats reversed all the observed effects . At the mRNA level , tie 2 was exclusively , but ang 1 predominantly , expressed in the stroma , compared with the epithelial compartment . Local delivery of soluble tie 2 during testosterone-stimulated growth , inhibited vascular maturation and increased vascular volume and leukocyte infiltration compared with controls . We conclude that androgens may regulate the prostate vasculature by direct effects on mural vascular cells and by influencing the secretion of the angiopoietins , in above all , the stroma cells . P10275 in nuclei of rat testis . Testis nuclei of hypophysectomized rats selectively accumulate labeled testosterone and 5alpha-dihydrotestosterone following the injection of tritiated testosterone in vivo . DB00624 and 5alpha-dihydrotestosterone are bound to macromolecules in nuclei and can be extracted with 0.5 M DB00761 . Accumulation of protein bound radioactive androgens in nuclei of isolated seminiferous tubules is similar to that of whole testis . The relative amounts of testosterone and dihydrotestosterone in purified nuclei were similar to the relative amounts bound to cytoplasmic receptors , suggesting that cytoplasmic androgen-receptor complexes may be transported into the nuclei . Binding of labeled androgen is saturable and inhibited by prior injection of unlabeled testosterone or cyproterone acetate . Nuclear binding sites are destroyed by the proteolytic enzyme pronase , but not by DNase . Like the cytoplasmic androgen-receptor complexes in rat testis , nuclear androgen-protein complexes are heat labile and dissociate slowly at 0 degrees C. androgens fail to accumulate in testis nuclei of the Stanley-Gumbreck androgen insensitive rat , a species lacking cytoplasmic androgen receptors in testis and other androgen target tissues . Different secretion patterns of matrix metalloproteinases and P10145 and effect of corticotropin-releasing hormone in preterm and term cervical fibroblasts . The aims of the present study were to compare the levels of mRNA and protein expression of matrix metalloproteinase ( MMP ) -1 , -3 , -8 and -9 in human cervical tissue in preterm and term labor as well as not in labor and to determine if corticotropin-releasing hormone ( P06850 ) has an effect on P03956 , -3 and interleukin ( IL ) -8 secretion in both preterm and term cervical fibroblasts . Cervical biopsies were taken from 60 women : 18 at preterm labor , 7 at preterm not in labor , 18 at term labor and 17 at term not in labor . ELISA and Immulite were used for protein and real-time RT-PCR for mRNA analysis . Cervical fibroblast cultures were incubated for 18 h with different P06850 concentrations ( 10(-13)-10(-6) M ) . The mRNA expression of P03956 , -3 and -9 was higher in laboring groups compared with term not in labor . Protein levels of P22894 and -9 were higher in term in labor group compared with non-laboring groups . There were no significant differences in mRNA and protein expression between the preterm and respective term control groups . P06850 significantly increased secretion of P10145 in preterm and term cervical fibroblasts compared with controls . The secretion of P10145 and P03956 was significantly higher and P08254 secretion lower in preterm cervical fibroblasts . In conclusion , cervical ripening at preterm seems to be a similar inflammatory process as at term with P06850 involved . However , preterm and term cervical fibroblasts might have different phenotypes based on different secretion patterns of P10145 , P03956 and P08254 . The androgen axis in recurrent prostate cancer . PURPOSE : Prostate cancer that recurs during androgen deprivation therapy is referred to as androgen-independent . High levels of expression of androgen receptor and androgen receptor-regulated genes in recurrent prostate cancer suggest a role for androgen receptor and its ligands in prostate cancer recurrence . EXPERIMENTAL DESIGN : Recurrent prostate cancer specimens from 22 men whose prostate cancer recurred locally during androgen deprivation therapy and benign prostate specimens from 48 men who had received no prior treatment were studied . P10275 expression was measured using monoclonal antibody and automated digital video image analysis . Tissue androgens were measured using radioimmunoassay . RESULTS : Epithelial nuclei androgen receptor immunostaining in recurrent prostate cancer ( mean optical density , 0.284 +/- SD 0.115 and percentage positive nuclei , 83.7 +/- 11.6 ) was similar to benign prostate ( mean optical density , 0.315 +/- 0.044 and percentage positive nuclei , 77.3 +/- 13.0 ) . Tissue levels of testosterone were similar in recurrent prostate cancer ( 2.78 +/- 2.34 pmol/g tissue ) and benign prostate ( 3.26 +/- 2.66 pmol/g tissue ) . Tissue levels of dihydrotestosterone , dehydroepiandrosterone , and androstenedione were lower ( Wilcoxon , P = 0.0000068 , 0.00093 , and 0.0089 , respectively ) in recurrent prostate cancer than in benign prostate , and mean dihydrotestosterone levels , although reduced , remained 1.45 nM . P10275 activation in recurrent prostate cancer was suggested by the androgen-regulated gene product , prostate-specific antigen , at 8.80 +/- 10.80 nmol/g tissue . CONCLUSIONS : DB00624 and dihydrotestosterone occur in recurrent prostate cancer tissue at levels sufficient to activate androgen receptor . Novel therapies for recurrent prostate cancer should target androgen receptor directly and prevent the formation of androgens within prostate cancer tissue . P10275 is widely expressed in bovine placentomes and up-regulated during differentiation of bovine trophoblast giant cells . OBJECTIVES : In cattle no biological role has been definitely identified for placental estrogens and progesterone . However , in the bovine trophoblast androgens may also be produced and have local effects . Thus , the aims of this study were to identify androgen receptor ( AR ) expressing cells and to monitor testosterone tissue concentrations in bovine placentomes throughout gestation . METHODS : Placental AR expression was characterized at the mRNA and protein level applying conventional and real-time RT-qPCR , western blot and immunohistochemistry . DB00624 was measured by radioimmunoassay . RESULTS : AR-mRNA was qualitatively detected from day 50 of gestation until term . Mean relative gene expression levels were constant between day 100 and late gestation . A slight non-significant increase was observed in the prepartal period . With immunohistochemistry distinct nuclear signals were predominantly observed in invasive trophoblast giant cells ( P21980 ) from day 80 until term . In mature P21980 of the trophoblast , immature P21980 and uninucleated trophoblast cells , stromal cells of the chorionic villi , caruncular epithelial and stromal cells immunoreactive score values were low at early and midgestation but increased significantly ( p < 0.01 ) during late gestation and remained high until parturition . With western blot in placentomal tissue a specific band of approximately 110 kDa was detected as it was the case in epididymis used as a positive control . DB00624 concentrations increased from 0.70 ± 0.29 pmol/g wet tissue between days 60-220 to 4.22 ± 1.29 pmol/g during late gestation ( p < 0.001 ) . DISCUSSION : The results are consistent with androgens as active products of bovine placental steroidogenesis . The substantial up-regulation of AR expression during P21980 differentiation suggests that androgens may be related to this process . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . Comparative characterization of lung muscarinic receptor binding after intratracheal administration of tiotropium , ipratropium , and glycopyrrolate . The aim of the current study was to characterize comparatively the binding of muscarinic receptor in the lung of rats intratracheally administered anticholinergic agents ( tiotropium , ipratropium , glycopyrrolate ) used clinically to treat chronic obstructive pulmonary disease ( P48444 ) and asthma . Binding parameters of [N-methyl-(3)H]scopolamine methyl chloride ( [(3)H] Q5H8A3 ) were determined in tissues ( lung , bladder , submaxillary gland ) of rats intratracheally administered tiotropium , ipratropium , and glycopyrrolate . The in vitro binding affinity of tiotropium for the receptors was 10-11-fold higher than those of ipratropium and glycopyrrolate . Intratracheal administration of tiotropium ( 0.6-6.4 nmol/kg ) caused sustained ( lasting at least 24 h ) increase in the apparent dissociation constant ( K(d) ) for [(3)H] Q5H8A3 binding in rat lung compared with the control value . Concomitantly , there was a long-lasting decrease in the maximal number of binding sites ( B(max) ) for [(3)H] Q5H8A3 . Similary , ipratropium and glycopyrrolate at 7.3 and 7.5 nmol/kg , respectively , brought about a significant increase in K(d) for [(3)H] Q5H8A3 binding . The effect by ipratropium was observed at 2 h but not 12 h , and that by glycopyrrolate lasted for 24 h . Both agents had little influence on the muscarinic receptors in the bladder and submaxillary gland . The present study provides the first evidence that tiotropium , ipratropium , and glycopyrrolate administered intratracheally in rats selectively bound muscarinic receptors of the lung , and tiotropium and glycopyrrolate had a much longer-lasting effect than ipratropium . Serum testosterone plays an important role in the metastatic ability of castration resistant prostate cancer . PURPOSE : Prostate cells are dependent on androgens for growth and proliferation . Androgen deprivation therapy is the recommended treatment for advanced/metastatic prostate cancer . Under this therapy , prostate cancer will inevitably progress to castration resistant prostate cancer ( CRPC ) . Despite putative castration resistance , testosterone might still play a crucial role in the progression of CRPC . The goal of this study was to determine the role of testosterone in the formation of metastases of CRPC in both in vitro and in vivo settings . METHODS : In vitro , the effect of testosterone and the non-aromatizable androgen methyltrienolone on migration , invasion and proliferation of a castration-resistant prostate cancer rat cell line ( Dunning R3327-MATLyLu ) was assessed using a transwell assay and a sulforhodamine B assay and immunohistochemical detection of ki67 . P10275 status was determined using Western blot . In vivo , Copenhagen rats were divided in four groups ( males , females , castrated males and females with testosterone suppletion ) and inoculated with MATLyLu cells . Tumor size was assessed daily . RESULTS : DB00624 increased cell migration and invasion in a concentration-dependent manner in vitro . DB00624 did not affect in vitro cell proliferation . No difference was shown between the effect of testosterone and methyltrienolone . In vivo , in groups with higher levels of circulating testosterone , more rats had (micro)metastases compared with groups with low levels of testosterone . No effect was observed on primary tumor size/growth . CONCLUSIONS : Despite assumed castration resistance , progression of prostate cancer is still influenced by androgens . Therefore , continuous suppression of serum testosterone in patients who show disease progression during castration therapy is still warranted . P10275 dependent and independent activities of testosterone on hepatic microsomal drug metabolism . Administration of testosterone for 6 days to intact female and castrate male BALB/cJ mice stimulated hepatic microsomal ethylmorphine N-demethylase activity and cytochrome P-450 content by 50-75 % . DB00624 also stimulated hepatic microsomal NADPH-oxidase activity , but to a lesser degree . To probe the mechanism of this effect of androgens , two antiandrogens ( cyproterone acetate and flutamide ) were employed . Since cyproterone acetate was a potent stimulator of hepatic microsomal ethylmorphine N-demethylase activity and cytochrome P-450 content , no antiandrogenic activity of this steroid could be detected . By contrast , flutamide alone had little effect on either ethylmorphine N-demethylase activity or cytochrome P-450 content . However , this drug effectively blocked the stimulatory effects of testosterone on ethylmorphine N-demethylase activity and cytochrome P-450 content but not on NADPH-oxidase activity . This effect was not species specific , since flutamide also prevented androgen stimulation of ethylmorphine metabolism in adult castrate and prepubertal male Fisher rats . The testosterone-induced increase of hepatic weight and microsomal protein content was not affected by the administration of flutamide . The observations are consistent with the hypothesis that androgens have two distinct effects on the liver . First , testosterone may act as a general , nonspecific stimulant of liver weight and microsomal protein content which is independent of the androgen receptor . Secondly , testosterone action in the liver may be expressed via an androgen-specific or androgen receptor-dependent mechanism which controls , in part , the cytochrome P-450-dependent demethylase system . Recent androgen receptor antagonists in prostate cancer . P10275 has been shown to promote prostate cell growth and carcinogenesis of prostate cancer by up-regulating its target genes . DB00624 and dihydrotestosterone are two major hormones which bind to and activate androgen receptor . Targeting both the androgen receptor and the enzymes catalyzing the biosynthesis of testosterone and dihydrotestosterone has been shown to be clinically beneficial in the treatment of prostate cancer . Prostate cancer can become castration-resistant after long term treatment with chemo drugs , so efforts in finding compounds with improved efficiency to castration-resistant prostate cancer are urgently needed . In this review we summarized the studies on recent progress in the development of small molecular AR antagonists for the treatment of prostate cancer . P10275 in human skin cytosol . Human skin , an accessible tissue , is an androgen target organ . We have measured the androgen-binding capacity of human skin cytosol using either 5 alpha[3H]dihydrotestosterone ( [3H] DB02901 ) or [3H]methyltrienolone ( [3H]R-1881 ) as ligand . Samples were incubated for 20 h at 0 C , and dextran-coated charcoal was used to separate bound from free steroids . The androgen receptor has a high affinity for both ligands ( 0.23 +/- 0.04 nM for [3H] DB02901 ; 0.32 +/- 0.16 nM for [3H]R-1881 ) . DB00624 , cyproterone acetate , and , to a lesser extent , estradiol also bind this protein . Progesterone displaces R-1881 from its binding sites , whereas its 5 alpha-reduced metabolite somewhat inhibits DB02901 binding . The highest binding capacity is measured in cytosol of skin from external genitalia ( 129.14 +/- 58.0 fmol/g skin ; n = 34 ) ; it is lower in pubic skin ( 21.8 +/- 13 fmol/g skin ; n = 6 ) . There is no variation as a function of age or sex in genital skin ; the higher concentrations observed in the cytosol of pubic skin of women compared to that of men are probably related to lower levels of endogenous steroids . Whereas most patients with the complete form of the testicular feminization syndrome do not have detectable concentrations of androgen receptor , one patient with apparent complete clinical androgen insensitivity had a normal androgen-binding capacity . The parity of values in genital skin from men and women , the absence of variation with age , and the presence of a cytosolic androgen receptor in some androgen-insensitive patients suggest that the androgen receptor in human skin cytosol is not regulated by androgens . P10275 expressing neurons that project to the paraventricular nucleus of the hypothalamus in the male rat . Androgen receptors are distributed throughout the central nervous system and are contained by a variety of nuclei that are known to project to or regulate the paraventricular nucleus ( PVN ) of the hypothalamus , the final common pathway by which the brain regulates the hypothalamic-pituitary-adrenal ( Q9Y251 ) response to homeostatic threat . Here we characterized androgen receptor staining within cells identified as projecting to the PVN in male rats bearing iontophoretic or crystalline injections of the retrograde tracer FluoroGold aimed at the caudal two-thirds of the nucleus , where corticotropin-releasing hormone-expressing neurons are amassed . P10275 ( AR ) and FluoroGold ( FG ) double labeling was revealed throughout the limbic forebrain , including scattered numbers of cells within the anterior and posterior subdivisions of the bed nuclei of the stria terminalis ; the medial zone of the hypothalamus , including large numbers of AR-FG-positive cells within the anteroventral periventricular and medial preoptic cell groups . Strong and consistent colabeling was also revealed throughout the hindbrain , predominantly within the periaqueductal gray and the lateral parabrachial nucleus , and within various medullary cell groups identified as catecholaminergic , predominantly C1 and A1 neurons of the ventral medulla . These connectional data predict that androgens can act on a large assortment of multimodal inputs to the PVN , including those involved with the processing of various types of sensory and limbic information , and provide an anatomical framework for understanding how gonadal status could contribute to individual differences in Q9Y251 function . Nursing supports neonatal porcine testicular development . The lactocrine hypothesis suggests a mechanism whereby milk-borne bioactive factors delivered to nursing offspring affect development of neonatal tissues . The objective of this study was to assess whether nursing affects testicular development in neonatal boars as reflected by : ( 1 ) Sertoli cell number and proliferation measured by GATA-4 expression and proliferating cell nuclear antigen immunostaining patterns ; ( 2 ) Leydig cell development and steroidogenic activity as reflected by insulin-like factor 3 ( P51460 ) , and P450 side chain cleavage ( scc ) enzyme expression ; and ( 3 ) expression of estrogen receptor-alpha ( P03372 ) , vascular endothelial growth factor ( P15692 ) A , and relaxin family peptide receptor ( RXFP ) 1 . At birth , boars were randomly assigned ( n = 6-7/group ) to nurse ad libitum or to be pan fed porcine milk replacer for 48 h . Testes were collected from boars at birth , before nursing and from nursed and replacer-fed boars at 50 h on postnatal day ( P01160 ) 2 . Sertoli cell proliferating cell nuclear antigen labeling index increased ( P < 0.01 ) from birth to P01160 2 in nursed , but not in replacer-fed boars . Sertoli cell number and testicular GATA-4 protein levels increased ( P < 0.01 ) from P01160 0 to P01160 2 only in nursed boars . Neither age nor nursing affected testicular P51460 , P450scc , P03372 , or P15692 levels . However , testicular relaxin family peptide receptor 1 ( Q9HBX9 ) levels increased ( P < 0.01 ) with age and were greater in replacer-fed boars on P01160 2 . Results suggest that nursing supports neonatal porcine testicular development and provide additional evidence for the importance of lactocrine signaling in pigs . Integrative analysis of transcriptomics , proteomics , and metabolomics data of white adipose and liver tissue of high-fat diet and rosiglitazone-treated insulin-resistant mice identified pathway alterations and molecular hubs . The incidences of obesity and type 2 diabetes are rapidly increasing and have evolved into a global epidemic . In this study , we analyzed the molecular effects of high-fat diet ( HFD ) -induced insulin-resistance on mice in two metabolic target tissues , the white adipose tissue ( WAT ) and the liver . Additionally , we analyzed the effects of drug treatment using the specific PPARγ ligand rosiglitazone . We integrated transcriptome , proteome , and metabolome data sets for a combined holistic view of molecular mechanisms in type 2 diabetes . Using network and pathway analyses , we identified hub proteins such as P21912 and P53597 in WAT and deregulation of major metabolic pathways in the insulin-resistant state , including the TCA cycle , oxidative phosphorylation , and branched chain amino acid metabolism . Rosiglitazone treatment resulted mainly in modulation via Q07869 signaling and oxidative phosphorylation in WAT only . Interestingly , in HFD liver , we could observe a decrease of proteins involved in vitamin B metabolism such as Q6P996 and P00374 and the according metabolites . Furthermore , we could identify sphingosine ( Sph ) and sphingosine 1-phosphate ( SP1 ) as a drug-specific marker pair in the liver . In summary , our data indicate physiological plasticity gained by interconnected molecular pathways to counteract metabolic dysregulation due to high calorie intake and drug treatment . N-glucuronidation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) and N_hydroxy-PhIP by specific human UDP-glucuronosyltransferases . Glucuronidation is a major metabolic pathway in the biotransformation of many xenobiotics . Recent studies have shown that in humans , UDP-glucuronosyltransferase ( P78381 ) -mediated glucuronidation plays a critical role in the detoxification of food-borne carcinogenic heterocyclic amines . 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , the most abundant carcinogenic heterocyclic amine found in well-cooked meats , has been shown to be extensively glucuronidated in humans . To determine which P78381 isozymes are involved in the biotransformation of PhIP and the cytochrome P4501A2-mediated reactive intermediate N-hydroxy-PhIP , microsomes expressing human P22309 , -1A4 , -1A6 or -1A9 were incubated with PhIP and N-hydroxy-PhIP and the reaction products analyzed by HPLC and P19957 -MS . Incubations containing N-hydroxy-PhIP and P22309 expressing microsomes , with an apparent Km of 4.58 microM and a Vmax of 4.18 pmol/min/mg protein , had the highest capacity to convert N-hydroxy-PhIP to N-hydroxy-PhIP-N2-glucuronide . Microsomes expressing O60656 produced N-hydroxy-PhIP-N3-glucuronide at the highest rate with an apparent Km and Vmax of 3.73 microM and 4.07 pmol/min/mg , respectively . A third previously undefined glucuronide accounted for 31 % of the total glucuronides formed from the P22310 expressing microsomes . No glucuronide conjugates were detected from microsomes expressing P19224 . Incubations containing PhIP as substrate formed direct PhIP-glucuronides in microsomes expressing P22309 , P22310 and O60656 but at levels averaging 53-fold lower than when N-hydroxy-PhIP was used as the substrate . Knowing the glucuronidation capacity of the specific P78381 isozymes involved in PhIP and N-hydroxy-PhIP glucuronidation should help in determining the individual susceptibility to the potential cancer risk from exposure to PhIP . Gender-specific effects of endogenous testosterone : female alpha-estrogen receptor-deficient C57Bl/6J mice develop glomerulosclerosis . Young female mice on a C57Bl/6J ( B6 ) background are considered glomerulosclerosis ( GS ) -resistant but aging B6 mice develop mild GS . Estrogen deficiency accelerates while estrogen replacement retards GS in young sclerosis-prone oligosyndactyly mutant mice on an P04000 background . To explore the effects of sex hormones on glomerular structure and function in the context of gender and genetic background , we studied mice in which the estrogen-receptor ( ER ) genes alpha- or -beta were deleted ( alpha- or betaER knockout ( KO ) ) and crossed into the B6 background . We also studied ovariectomized ( Ovx ) B6 mice given testosterone . Male and female betaERKO and male alphaERKO mice had no glomerular dysfunction at 9 months of age ; however , alphaERKO female mice displayed albuminuria and GS . Ovx prevented glomerular dysfunction in alphaERKO female mice by eliminating endogenous testosterone production while exogenous testosterone induced GS in Ovx B6 mice . P10275 ( AR ) expression and function was found in microdissected glomeruli and cultured mesangial cells . DB00624 compared to placebo increased both AR expression and TGF-beta1 mRNA levels in glomeruli isolated from female B6 mice . Estrogen deficiency had no deleterious effects on the glomeruli in B6 mice . Our study shows that genetic traits strongly influence the GS-promoting effects of estrogen deficiency while testosterone induces GS in a gender-specific manner . DB01285 -releasing hormone inhibits insulin-like growth factor-I release from primary cultures of rat granulosa cells . DB01285 -releasing hormone ( P06850 ) , a neuropeptide which modulates gonadal function during stress , is expressed by several cell types of the rat ovary and is able to suppress oestrogen release from rat granulosa cells . The mechanism of this effect is , however , not known . Since insulin-like growth factor ( IGF ) -I is produced by rat granulosa cells and exerts a synergistic role with DB00094 on granulosa cell steroidogenesis , we hypothesised that P06850 may suppress oestrogen release from granulosa cells by inhibiting P05019 release and/or stimulating the release of its binding protein ( P17936 ) . To test this hypothesis , granulosa cells were obtained from immature female Sprague-Dawley rats primed with diethylstilboestrol , and hormone concentrations were measured in the conditioned medium by radioimmunoassay . P06850 suppressed oestrogen and P05019 release stimulated by DB00094 used at a concentration of 1 IU/l , whereas it did not have any statistically significant effect on oestrogen and P05019 release in basal conditions or in response to 5 IU/l DB00094 . The suppressive effects of P06850 on oestrogen and P05019 release were antagonised by a selective P06850 receptor antagonist . P06850 had no effects on P17936 release . P06850 did not have any effect on oestrogen release stimulated by increasing concentrations of P05019 and its suppressive effect on DB00094 -stimulated oestrogen release was overcome by the addition of low doses of exogenous P05019 . In conclusion , P06850 suppressed the release of oestrogen and P05019 , but not of P17936 . Thus , the inhibitory effects of P06850 on oestrogen release could be mediated , partly , by a suppression of the autocrine/paracrine action of P05019 . Maternal separation produces alterations of forebrain brain-derived neurotrophic factor expression in differently aged rats . Early life adversity , such as postnatal maternal separation ( MS ) , play a central role in the development of psychopathologies during individual ontogeny . In this study , we investigated the effects of repeated MS ( 4 h per day from postnatal day ( P01160 ) 1-21 ) on the brain-derived neurotrophic factor ( P23560 ) expression in the medial prefrontal cortex ( mPFC ) , the nucleus accumbens ( NAc ) and the hippocampus of male and female juvenile ( P01160 21 ) , adolescent ( P01160 35 ) and young adult ( P01160 56 ) Wistar rats . The results indicated that MS increased P23560 in the P00915 and the dentate gyrus ( DG ) of adolescent rats as well as in the DG of young adult rats . However , the expression of P23560 in the mPFC in the young adult rats was decreased by MS . Additionally , in the hippocampus , there was decreased P23560 expression with age in both the MS and non separated rats . However , in the mPFC , the P23560 expression was increased with age in the non separated rats ; nevertheless , the P23560 expression was significantly decreased in the MS young adult rats . In the NAc , the P23560 expression was increased with age in the male non-maternal separation ( Q5H8A3 ) rats , and the young adult female MS rats had less P23560 expression than the adolescent female MS rats . The present study shows unique age-differently changes on a molecular level induced by MS and advances the use of MS as a valid animal model to detect the underlying neurobiological mechanisms of mental disorders . The growth hormone dependent serine protease inhibitor , Spi 2.1 inhibits the des ( 1-3 ) insulin-like growth factor-I generating protease . The conversion of insulin-like growth factor-I ( P05019 ) to the biologically more active des ( 1-3 ) P05019 variant is catalyzed by a ubiquitous protease . This proteolytic activity is inhibited by human alpha1-antitrypsin and soy-bean trypsin inhibitor and is up-regulated in serum and tissue extracts of hypophysectomized rats . These observations lead us to investigate whether the growth hormone regulated , serine protease inhibitor , Spi 2.1 was able to inhibit the des ( 1-3 ) P05019 generating protease . P00374 deficient Chinese hamster ovary ( CHO(dhfr-ve) ) cells were transfected with a rat Spi 2.1 expression vector containing the dhfr and neomycin resistance gene . Stable transfectants were selected using G418 and amplified using methotrexate . Conditioned medium from Spi 2.1 transfected CHO cells potently inhibited proteolytic activity directed against a synthetic hexa-peptide with a sequence identical to the N-terminal of P05019 . In contrast conditioned medium from wild-type CHO cells had little effect . Based upon these observations we suggest that our previous finding of enhanced des ( 1-3 ) P05019 generating protease activity in growth hormone deficient rats may be , at least partly explained by reduced levels of Spi 2.1 . Furthermore , we propose that the regulation of the generation of des ( 1-3 ) P05019 may be an additional potential site of growth hormone regulation of P05019 action . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . Androgens and metabolic syndrome : lessons from androgen receptor knock out ( ARKO ) mice . DB00624 ( T ) is an important factor for determining body composition in males . Abdominal obesity is inversely correlated with serum T levels in men , leading to greater mortality . Pathologically hypogonadal men also have a significantly higher fat mass , which is reversed by T administration . However , the mechanism for such anti-obesity effect of androgen has not been well clarified . P10275 ( AR ) null male mice revealed late-onset obesity . Male ARKO mice were euphagic compared to the wild-type male controls , but also less dynamic and less oxygen consuming . Transcript profiling indicated that male ARKO mice had lower transcripts for the thermogenetic uncoupling protein 1 ( P25874 ) . We also found enhanced secretion of adiponectin , which is insulin-sensitizing , from adipose tissue in comparison to wild type , which might partly explain why the overall insulin sensitivity of male ARKO mice remained almost intact despite their apparent obesity . In addition , decreased lipolysis rather than increased lipid synthesis was observed , which might also account for the increased adiposity in male ARKO mice . The results revealed that AR plays important roles in male metabolism by affecting the energy balance , and is negative to both adiposity and insulin sensitivity . DB00624 modulation of dendritic spines of somatosensory cortical pyramidal neurons . Brain structures and functions are increasingly recognized to be directly affected by gonadal hormones , which classically determine reproductive functions and sexual phenotypes . In this regard , we found recently that ovariectomy trimmed the dendritic spines of female rat primary somatosensory cortical neurons and estradiol supplement reversed it . Here , we investigated whether in the male androgen also has a cortical modulatory effect . The dendritic arbors and spines of rat somatosensory cortical pyramidal neurons were studied following intracellular dye injection and three-dimensional reconstruction . Dendritic spines , but not length , of the layers III and V pyramidal neurons were found reduced at 2 weeks and rebounded slightly at 4 weeks and further at 8 and 24 weeks following castration , which , however , remained significantly fewer than those of the intact animals . Two weeks of osmotic pump-delivered testosterone treatment to animals castrated for 4 weeks replenished serum testosterone and reversed the densities of dendritic spines on these neurons to control animal levels . P10275 appears to mediate this effect as its antagonist flutamide reduced the dendritic spines of normal adult rats while causing a mild feedback surge of serum testosterone . On the other hand , blocking the conversion of testosterone to estrogen with the aromatase inhibitor anastrozole failed to alter the dendritic spine densities in male adult rats . In conclusion , these results support our hypothesis that testosterone acts directly on the androgen receptor in males to modulate the dendritic spines of somatosensory cortical output neurons . Androgen receptors , serum androgen levels and survival of breast cancer patients . Steroid receptor levels and serum androgen levels were determined in 61 breast cancer patients and 34 patients with non-malignant breast lesions . DB00624 and dehydroepiandrosterone-sulfate did not and androstenedione did show a difference between the two groups . Androgen levels had no influence on survival rates . P10275 ( AR ) levels correlated with progesterone receptor levels , but not with estrogen receptor levels or with tumor stage . Patients with positive AR findings had a better survival rate ; this was independent of tumor stage . AR findings may therefore be a prognostic index in breast cancer patients . P10275 -immunoreactive cells in ram hypothalamus : distribution and co-localization patterns with gonadotropin-releasing hormone , somatostatin and tyrosine hydroxylase . DB00624 exerts important feedback effects on the hypothalamus of the ram to influence reproductive functioning . To provide a neuroanatomical basis for understanding this androgen action , the present study has examined androgen receptor ( AR ) immunoreactivity within the hypothalamus and adjacent brain areas of the intact non-breeding season ram . The largest populations of AR-immunoreactive cells were detected in the medial preoptic area , infundibular and premammillary nuclei in addition to the ventromedial nucleus ( VMN ) where cells were found distributed throughout its medial and lateral divisions . Smaller numbers of AR-expressing cells were identified in the bed nucleus of the stria terminalis and anterior hypothalamic area ( DB00551 ) including the paraventricular , but not the supraoptic , nucleus . Double-labelling immunocytochemistry revealed the presence of AR immunoreactivity in only 2 of 460 gonadotropin-releasing hormone ( DB00644 ) neurons . A very small population of TH-immunoreactive cells located in the lateral aspect of the DB00551 was found to contain ARs . Dopaminergic cells elsewhere in the hypothalamus , including the infundibular nucleus , did not display AR immunoreactivity . Nearly 50 % of AR-expressing cells in the lateral VMN were immunoreactive for somatostatin while less than 5 % of periventricular somatostatin neurons displayed AR immunoreactivity . These results show where ARs are expressed in the ram hypothalamus and indicate the neuroanatomical sites at which androgen may act to influence reproductive function . The absence of ARs in the neuroendocrine DB00644 and tuberoinfundibular dopaminergic cells suggests that androgens do not influence the genome of these cells in any direct manner . In contrast , the somatostatin neurons of the VMN appear to be an important target for circulating androgens in the non-breeding season ram . Pubertal maturation is associated with an increase in the number of androgen receptor-immunoreactive cells in the brains of male ferrets . P10275 -immunoreactive ( AR-IR ) cells were identified in brains of male ferrets before and after the onset of pubertal maturation . There was a greater number of AR-IR cells after the onset of pubertal maturation in some , but not all , brain regions examined . Regions in which the number of AR-IR cells increased included the preoptic area and the amygdala , areas known to be involved in the control of male reproductive behaviors . The mechanisms responsible for the increase in AR-IR cells are unknown , but might be related to the higher circulating levels of testosterone that were present in the older animals . DB00624 may increase androgen receptor ( AR ) immunoreactivity both by concentrating already existing ARs within the nucleus and by stimulating de novo synthesis of receptor protein . Comparison of the effects of cytoprotective drugs on human plasma adrenocorticotropic hormone and cortisol levels with continual stress exposure . Cetraxate hydrochloride ( cetraxate ) , ecabet sodium ( ecabet ) , and sulpiride , which are cytoprotective drugs , have been used to treat peptic ulcers and acute or chronic gastritis . They are reported to improve mucosal blood flow in the stomach . One of the most important factors believed to cause gastric ulcers is mental and/or physiological stress . When people feel stress , the hypothalamo-pituitary-adrenal ( Q9Y251 ) axis is activated . Therefore , corticotropin-releasing hormone ( P06850 ) , adrenocorticotropic hormone ( DB01285 ) , and cortisol can be indicators of stress . We examined the effects of cetraxate , ecabet and sulpiride on the plasma levels of DB01285 and cortisol under stress conditions by repetitive blood sampling . Venous blood samples were taken before and 20-240 min after a single administration of the drugs or a placebo . A single dose of ecabet caused significant suppression of increases in plasma DB01285 -like immunoreactive substance ( IS ) levels at 90 to 120 min and cortisol levels at 240 min , compared with the response to placebo . DB00391 only suppressed increases in plasma cortisol levels at 180 to 240 min , compared with the response to placebo . A single dose of cetraxate had no effect on plasma DB01285 -IS and cortisol levels . Ecabet may have a modulatory effect on the Q9Y251 axis while sulpiride may have a partial modulatory effect on the Q9Y251 axis . These effects might be beneficial in stress-related disease . 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 . Serotonin transporter , sex , and hypoxia : microarray analysis in the pulmonary arteries of mice identifies genes with relevance to human PAH . Pulmonary arterial hypertension ( PAH ) is up to threefold more prevalent in women than men . Female mice overexpressing the serotonin transporter ( P31645 ; P31645 + mice ) exhibit PAH and exaggerated hypoxia-induced PAH , whereas male P31645 + mice remain unaffected . To further investigate these sex differences , microarray analysis was performed in the pulmonary arteries of normoxic and chronically hypoxic female and male P31645 + mice . Quantitative RT-PCR analysis was employed for validation of the microarray data . In relevant groups , immunoblotting was performed for genes of interest ( CEBPβ , Q16678 , and P01100 ) . To translate clinical relevance to our findings , CEBPβ , Q16678 , and P01100 mRNA and protein expression was assessed in pulmonary artery smooth muscle cells ( PASMCs ) derived from idiopathic PAH ( IPAH ) patients and controls . In female P31645 + mice , multiple pathways with relevance to PAH were altered . This was also observed in chronically hypoxic female P31645 + mice . We selected 10 genes of interest for qRT-PCR analysis ( P01100 , CEBPβ , Q16678 , P08590 , HAMP2 , P02788 , P26678 , P01160 , P25874 , and P09871 ) , and 100 % concordance was reported . Protein expression of three selected genes , CEBPβ , Q16678 , P01100 , was also upregulated in female P31645 + mice . Serotonin and 17β-estradiol increased CEBPβ , Q16678 , and P01100 protein expression in PASMCs . In addition , CEBPβ , Q16678 , and P01100 mRNA and protein expression was also increased in PASMCs derived from IPAH patients . Here , we have identified a number of genes that may predispose female P31645 + mice to PAH , and these findings may also be relevant to human PAH . P10275 gene polymorphisms in amyotrophic lateral sclerosis . Amyotrophic lateral sclerosis ( P35858 ) is more common in men than in women ( male to female ratio of approximately 2:1 ) , suggesting a role for a sex-linked factor in the disease . The recent identification of a mutation of the androgen receptor gene in Kennedy 's disease or X-linked bulbospinal neuronopathy , a rare form of progressive lower motor neurone degeneration , also associated with clinical signs of androgen insensitivity , raises the possibility that androgen function may be disturbed in other motor neurone disorders , including P35858 . The Kennedy 's disease mutation consists of an increased size of a highly polymorphic CAG repeat sequence in the first exon of the androgen receptor gene , coding for a polyglutamine tract . We have analysed this CAG repeat sequence in a large number of patients with typical sporadic P35858 and in normal controls , in order to test the hypothesis that this polymorphism of the androgen receptor gene may influence susceptibility for P35858 . We report that the distribution of alleles relating to the size of the CAG repeat sequence of the androgen receptor gene is similar in P35858 and controls , indicating that polymorphisms of the CAG repeat sequence of the androgen receptor gene play a limited role , if any , in susceptibility to P35858 . Increased dihydrotestosterone receptor levels in high-stage renal adenocarcinoma . Primary renal adenocarcinoma tissue , metastatic deposits , and normal kidney parenchyma from 16 patients were assayed for sex hormone receptors by dextran-coated charcoal adsorption and sucrose gradient centrifugation techniques . DB02901 receptors ( P10275 ) were found in all renal carcinomatous tissue ( 20/20 ) and in 93 % ( 13/14 ) autologous normal kidneys analyzed . DB00624 receptors were found in 84 % ( 16/19 ) of tumors and 93 % ( 14/15 ) or normal kidneys analyzed . Estrogen receptors in small amounts ( ER ) were detected in only 5 % ( 1/19 ) of tumors and in 7 % ( 1/15 ) of normal kidneys . Progesterone receptors ( PR ) in low quantities were detected in 30 % ( 6/20 ) of renal tumors and in 40 % ( 6/15 ) of normal kidneys . P10275 levels in high-stage tumors ( DB00279 , DB00451 ) were significantly elevated over levels in autologous normal kidney , whereas in low-stage tumors localized to the kidney ( T1 and P24752 tumors ) P10275 levels were not significantly different from autologous normal kidney . The mean levels of P10275 in high-stage kidney tumors were significantly elevated over levels in low-stage tumors ( P less than 0.001 ) . P10275 estimation in renal neoplasms may help in biologic staging of renal adenocarcinoma and could define a group of patients in whom anti-androgen therapy may be worth a trial . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . P10275 -dependent transactivation of growth arrest-specific gene 6 mediates inhibitory effects of testosterone on vascular calcification . Recent epidemiological studies have found that androgen deficiency is associated with a higher incidence of cardiovascular disease in men . However , little is known about the mechanism underlying the cardioprotective effects of androgens . Here we show the inhibitory effects of testosterone on vascular calcification and a critical role of androgen receptor ( AR ) -dependent transactivation of growth arrest-specific gene 6 ( Gas6 ) , a key regulator of inorganic phosphate ( P(i) ) -induced calcification of vascular smooth muscle cells ( VSMC ) . DB00624 and nonaromatizable androgen dihydrotestosterone inhibited P(i)-induced calcification of human aortic VSMC in a concentration-dependent manner . Androgen inhibited P(i)-induced VSMC apoptosis , an essential process for VSMC calcification . The effects on VSMC calcification were mediated by restoration of P(i)-induced down-regulation of Gas6 expression and a subsequent reduction of Akt phosphorylation . These effects of androgen were blocked by an AR antagonist , flutamide , but not by an estrogen receptor antagonist , DB00947 . We then explored the mechanistic role of the AR in Gas6 expression and found an abundant expression of AR predominantly in the nucleus of VSMC and two consensus ARE sequences in the Gas6 promoter region . DB02901 stimulated Gas6 promoter activity , and this effect was abrogated by flutamide and by AR siRNA . Site-specific mutation revealed that the proximal ARE was essential for androgen-dependent transactivation of Gas6 . Furthermore , chromatin immunoprecipitation assays demonstrated ligand-dependent binding of the AR to the proximal ARE of Gas6 . These results indicate that AR signaling directly regulates Gas6 transcription , which leads to inhibition of vascular calcification , and provides a mechanistic insight into the cardioprotective action of androgens . Novel function of androgen receptor-associated protein 55/ O43294 as a negative regulator of P84022 signaling . P10275 -associated protein 55 ( O43294 / O43294 ) belongs to the LIM protein superfamily and is featured by three or four N-terminal LD motifs and four C-terminal zinc finger-like LIM domains . Both LD motifs and LIM domains can serve as protein-protein interaction interfaces . Recently , we found that enforced expression of O43294 inhibits transforming growth factor-beta-mediated up-regulation of Smad binding element-luciferase reporter activity in NRP-154 and NRP-152 rat prostate and LNCaP human prostate cell lines . Moreover , O43294 also inhibits the induction of Smad-binding element 4-luciferase and 3TP-luciferase ( a plasminogen activator inhibitor-1 ( P05121 ) promoter construct ) reporters by constitutively active ( CA ) - P84022 in these cell lines . Co-immunoprecipitation studies suggest an interaction between O43294 and either CA- P84022 or wild-type P84022 in HEK293 cells that occurs through the MH2 domain of P84022 and the C terminus of O43294 with wild-type P84022 having stronger affinity than CA- P84022 to O43294 . O60760 pull-down assays demonstrate that this interaction can occur in a cell-free system . These results are consistent with the luciferase data showing that the C terminus of O43294 is critical for suppression of P84022 activity . Furthermore , using a mammalian two-hybrid system , we confirmed that O43294 interacts with the MH2 domain of P84022 and suppresses CA- P84022 -induced transcriptional responses . In conclusion , these results support that O43294 selectively intercepts transforming growth factor-beta signaling through an interaction of the LIM domain of O43294 with the MH2 domain of P84022 . P10275 signals regulate UDP-glucuronosyltransferases in the urinary bladder : a potential mechanism of androgen-induced bladder carcinogenesis . UDP-glucuronosyltransferases ( UGTs ) , major phase II drug metabolism enzymes , play an important role in urinary bladder cancer initiation by detoxifying carcinogens . We aimed to determine if androgens regulate P78381 expression via the androgen receptor ( AR ) pathway in the bladder . Real-time reverse transcription-polymerase chain reaction and Western blot analyses were used to assess P22309 levels in the normal urothelium SVHUC cell line stably expressed with AR and in bladder tissues from AR knockout ( ARKO ) and castrated male mice . Immunohistochemistry was also performed in radical cystectomy specimens . DB02901 ( DB02901 ) treatment in SVHUC-AR reduced mRNA expression of all the P22309 subtypes ( 19-75 % decrease ) , and hydroxyflutamide antagonized the DB02901 effects . In contrast , DB02901 showed only marginal effects on P22309 expression in SVHUC-Vector . Of note were higher expression levels of UGT1As in SVHUC-Vector than in SVHUC-AR . In ARKO mice , all the Ugt1a subtypes were up-regulated , compared to wild-type littermates . In wild-type male mice , castration increased the expression of Ugt1a8 , Ugt1a9 , and Ugt1a10 . Additionally , wild-type female mice had higher levels of Ugt1a than wild-type males . Immunohistochemical studies showed strong ( 3+ ) P22309 staining in 11/24 ( 46 % ) cancer tissues , which was significantly lower than in corresponding benign tissues [ 17/18 ( 94 % ) cases ( P = 0.0009 ) ] . These results suggest that androgen-mediated AR signals promote bladder carcinogenesis by down-regulating the expression of UGTs in the bladder . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . P10275 signaling induced by supraphysiological doses of dihydrotestosterone in human peripheral blood lymphocytes . Anabolic androgenic steroids , a class of steroid hormones related to testosterone , are natural ligands of androgen receptor ( AR ) , a member of the nuclear receptor superfamily of ligand-activated transcription factors . AR binds specific DNA elements , known as androgen-response elements . DB00624 , the main male sexual hormone , binds AR directly and indirectly , through conversion into dihydrotestosterone ( DB02901 ) , its more active metabolite . Anabolic androgenic steroids are frequently detected in the urine of doped athletes ; their consumption is also growing among sport amateurs and adolescents . The effects of androgens can differ depending on the target cells and/or tissues . To gain insight into transcription activation mechanisms of AR , we investigated AR protein signaling in human peripheral blood lymphocytes treated with supraphysiological doses of DB02901 . We performed a comparative proteomic analysis and we identified about 30 differentially expressed proteins . At least five species contained a consensus androgen-response elements sequence in the promoter region of related coding genes . The analysis also revealed that high doses of DB02901 activate the drug detoxification process , could stimulate an increase in cell motility and exert a prosurvival effect rather than an apoptotic one . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . DB00624 inhibits matrix metalloproteinase-1 production in human endometrial stromal cells in vitro . P10275 ( AR ) is reported to be expressed in human uterine endometrium , but not much information is available on the role of androgens in human endometrium . The purpose of this study is to investigate the role of androgens in the regulation of matrix metalloproteinase ( MMP ) -1 , which is one of the important MMPs for menstruation and embryo implantation in human endometrium . Human endometrial stromal cells ( HESCs ) were obtained from human endometrium by enzymatic dissociation method . Purified HESCs were incubated with 17beta-estradiol ( E2 ) , testosterone , or E2 + testosterone . Progestins ( natural progesterone or medroxyprogesterone acetate ) or vehicle ( dimethyl sulfoxide ) were also added to the media instead of testosterone . Furthermore , hydroxyflutamide (FLU),a specific AR antagonist , was also supplemented to cultured media . The amounts of P03956 in cultured media and in HESC lysates were examined by ELISA measurements and western blotting analysis respectively . The expression of ARmRNA in HESCs RNA was analyzed by RT-PCR . DB00624 significantly inhibited P03956 in both cultured media and cell lysates in a dose-dependent manner . Progestins also inhibited P03956 . Furthermore , FLU completely recovered the decrease of P03956 induced by testosterone . ARmRNA was detected in all HESCs RNA . The present study demonstrated that the secretion and production of P03956 in HESCs in vitro were inhibited by testosterone through androgen receptors in a manner similar to that seen for progesterone . These findings indicate that androgen may play an important role in morphological and functional changes of human endometrium . P10275 up-regulates insulin-like growth factor binding protein-5 ( P24593 ) expression in a human prostate cancer xenograft . The insulin-like growth factor ( IGF ) binding proteins ( IGFBPs ) are important modulators of IGF action in many tissues including human prostate . IGFBPs and the androgen receptor ( AR ) are expressed in CWR22 , an androgen-dependent epithelial cell human CaP xenograft that retains biological characteristics of human CaPs , including regression following androgen withdrawal and recurrent growth of AR-containing cells in the absence of testicular androgens beginning several months after castration . Northern blot and in situ hybridization analyses demonstrated that P24593 is androgen-regulated in CWR22 . P24593 messenger RNA ( mRNA ) decreased by 90 % following castration of tumor-bearing mice compared with noncastrate androgen-stimulated mice . DB00624 treatment of CWR22 tumor-bearing mice 6 or 12 days after castration increased P24593 mRNA 10- to 12-fold . Levels of other IGFBP mRNAs did not change following androgen withdrawal and replacement . P24593 protein in tumor extracts bound 125I-labeled P05019 in ligand blot assays and the amounts of P24593 measured by immunoblotting paralleled the levels of P24593 mRNA . Androgen-induced expression of P24593 was at a maximum level within 24 h after testosterone replacement , whereas the major increase in cell proliferation as measured by Ki-67 immunostaining occurred between 24-48 h . This time course suggested P24593 may be a mediator of androgen-induced growth of CWR22 . In tumors that recurred several months following castration , P24593 mRNA and protein increased to levels that approached those in androgen-stimulated CWR22 tumors from noncastrate mice . P24593 immunohistochemical staining of prostate tissue specimens from patients was stronger in androgen-dependent and androgen-independent CaP than in areas of intraepithelial neoplasia ( P63167 ) or benign prostatic hyperplasia ( BPH ) . P24593 mRNA in these specimens was localized predominantly to stromal cells and P24593 protein to epithelial cell membranes . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . Targeting androgen receptor in estrogen receptor-negative breast cancer . Endocrine therapies for breast cancer that target the estrogen receptor ( ER ) are ineffective in the 25 % -30 % of cases that are ER negative ( ER- ) . P10275 ( AR ) is expressed in 60 % -70 % of breast tumors , independent of ER status . How androgens and AR regulate breast cancer growth remains largely unknown . We find that AR is enriched in ER- breast tumors that overexpress P04626 . Through analysis of the AR cistrome and androgen-regulated gene expression in ER-/ P04626 + breast cancers we find that AR mediates ligand-dependent activation of Wnt and P04626 signaling pathways through direct transcriptional induction of P56706 and P21860 . Specific targeting of AR , Wnt or P04626 signaling impairs androgen-stimulated tumor cell growth suggesting potential therapeutic approaches for ER-/ P04626 + breast cancers . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . P10275 is responsible for rat organic cation transporter 2 gene regulation but not for rOCT1 and rOCT3 . PURPOSE : Organic cation transporters 1-3 ( OCT1-3 ; Slc22a1-3 ) mediate the membrane transport of organic cations in the kidney . We previously reported that rat (r)OCT2 expression in the kidney was regulated by testosterone . In this study , we examined the transcriptional mechanisms underlying the testosterone-dependent regulation of rOCT2 expression . METHODS : Approximately 3000-bp fragments of the rOCT1-3 promoter region were isolated , and promoter activities were measured in the renal epithelial cell line LLC- P30613 with the coexpression of rat androgen receptor . RESULTS : Among reporter constructs tested , only rOCT2 promoter activity was stimulated by testosterone . This stimulation was suppressed by nilutamide , an antiandrogen drug . Reporter assays using deletion constructs and mutational constructs of putative androgen response elements ( ARE ) in the rOCT2 promoter region suggested that two AREs , located at approximately -3000 and -1300 , respectively , play an important role in the induction by testosterone . CONCLUSIONS : DB00624 induces the expression of rOCT2 , but not of rOCT1 and rOCT3 , via the AR-mediated transcriptional pathway . This is the first study to address the transcriptional mechanisms of testosterone-dependent gene regulation of the Slc22 family . P10275 (CAG)n polymorphism and androgen levels in women with systemic lupus erythematosus and healthy controls . Systemic lupus erythematosus ( SLE ) is an autoimmune disorder that affects mainly females . Therefore , interrelations between the reproductive and immune system have been assumed . Considering the complex influence of hormones and receptors , we aimed to investigate the influence of androgens and androgen receptor ( AR ) polymorphism in women with SLE . One hundred and sixteen patients and 44 healthy women were investigated . DB00624 , sex hormone-binding globulin ( P04278 ) , dehydroepiandrosterone-sulphate ( DHEAS ) concentrations and AR (CAG)n polymorphism were determined . SLE patients had significantly lower levels of total and free testosterone and DHEAS in comparison with the controls . No differences in the CAG repeat length between the groups were established . Women with two alleles carrying more than 22 CAG repeats had significantly higher levels of P04278 ( 101.51 ± 61.81 vs. 69.22 ± 45.93 nmol/l , p = 0.015 ) and DHEAS ( 3.11 ± 2.65 vs. 2.11 ± 3.06 μmol/l , p = 0.007 ) and a tendency to higher testosterone concentrations ( 2.35 ± 2.10 vs. 1.71 ± 1.70 nmol/l , p = 0.056 ) in comparison with other women . The CAG repeat length in the relatively longer (CAG)n allele was inversely related to the Systemic Lupus International Collaborating Clinics/ P10323 index ( r = -0.258 , p = 0.009 ) . In conclusion , the androgen receptor (CAG)n polymorphism is not related to the development of SLE , but it could modulate the severity of the lupus chronic damages as well as the androgen levels in women . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Delayed haemolytic transfusion reaction caused by anti-M antibody in a patient receiving interleukin-2 and interferon for metastatic renal cell cancer . Anti-M is usually a naturally occurring cold-reactive immunoglobulin M ( IgM ) antibody , often with an immunoglobulin G ( IgG ) component , and is seldom implicated in delayed haemolytic transfusion reactions ( P10275 ) . However , cases have been reported . In the majority , a P10275 is not suspected until further blood is requested and a new antibody is detected on pretransfusion testing . We describe the case of a young man receiving therapy with interleukin-2 ( P60568 ) and interferon-alpha ( IFN-alpha ) for metastatic renal cell cancer who developed a clinically suspected P10275 that was confirmed serologically to be caused by anti-M , reactive at 37 degrees C . We discuss the possible role of his biochemotherapy in the development of the P10275 . P10275 negatively influences the expression of chemokine receptors ( P61073 , P32246 ) and ligand-mediated migration in prostate cancer DU-145 . We previously reported that androgen receptor ( AR ) plays a role in the regulation of adhesion to the extracellular matrix and invasion of human prostate cancer cells by influencing the expression of specific integrin subunits . It is now considered that chemokines play a significant role in organ-selective cancer metastasis . In this study , we hypothesized that AR may influence the expression of these chemokine receptors and cell function . The mRNA expression of chemokine receptors in human prostate cancer cell line DU-145 and DU-145 cells expressing AR ( DU-145/AR ) was investigated by RT-PCR . DU-145 cells selectively expressed P61073 and P32246 mRNA at high levels compared with DU-145/AR cells . DU-145 showed vigorous migratory responses to its ligand P48061 ( also called stromal-derived factor-1alpha , SDF-1alpha ) and P10147 ( also called macrophage inflammatory protein-1 , MIP-1alpha ) . In contrast , neither P48061 nor P10147 affected the migration of DU-145/AR cells . These results indicate that expression of AR down-regulates the migratory responses of human prostate cancer cells via chemokine and its receptor systems . 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 . P10275 is essential for sexual differentiation of responses to olfactory cues in mice . During sexual differentiation males and females are exposed to different levels of testosterone , which promotes sex differences in the adult brain and in behavior . DB00624 can act after aromatization or reduction via a number of steroid hormone receptors . Here we provide new evidence that the androgen receptor ( AR ) is essential for sexual differentiation in mice . We used mice carrying the testicular feminization ( Tfm ) mutation of the AR . Adult Tfm males , wild-type male and female littermates were gonadectomized and given subcutaneous estradiol implants . In all sexually dimorphic traits , Tfm males had responses equivalent to females and different from males . In simultaneous choice tests , males spent significantly more time investigating female-soiled bedding , whereas females and Tfm males preferred to investigate male-soiled bedding . Tfm males and females did not have a partner preference in tests with awake stimulus animals , whereas males showed a preference for females over males . Exposure to male-soiled , but not clean , bedding produced a significant increase in c-Fos-immunoreactive cells in the medial preoptic area and bed nucleus of the stria terminalis in Tfm males and females , no increase was noted in males . Masculine sexual behavior ( mounting and thrusting ) was not sexually dimorphic , and all groups displayed these behaviors . Our results support data collected in humans suggesting a role for the androgen receptor in sexual differentiation of social preferences and neural responses to pheromones .