interacts_with DB01285?
Stress-induced corticotropin-releasing hormone-mediated P59044 inflammasome inhibition and transmissible enteritis in mice . BACKGROUND & AIMS : Stress alters brain-gut interactions and could exacerbate intestinal disorders , including irritable bowel syndrome . Alterations in the intestinal microbiota have been associated with irritable bowel syndrome . Maintenance of healthy microbiota requires nucleotide-binding oligomerization domain protein-like receptors , pyrin-domain containing ( NLRP ) -6 inflammasomes . We investigated the involvement of P59044 in water-avoidance stress ( P42768 ) -induced intestinal disorders in mice . METHODS : B57BL6 mice were subjected to P42768 for 1 hour each day for 10 days ; body weights and intestinal inflammation and permeability were analyzed . We investigated signaling via the Q96P20 and P59044 inflammasomes , and the role of corticotropin-releasing hormone ( P06850 ) in P42768 -associated inflammation and P59044 inhibition . Mice that were not exposed to stress were co-housed with mice subjected to P42768 to determine the effects of P42768 -induced dysbiosis , measured by sequencing bacterial 16S ribosomal RNA . We also assessed the effects of a peroxisome proliferator-activated receptor-γ agonist and probiotics . RESULTS : P42768 -induced small-bowel inflammation ( enteritis ) was associated with inhibition of P59044 , but not Q96P20 , and was prevented by a peroxisome proliferator-activated receptor-γ agonist , which induced epithelial expression of P59044 . P06850 was released during P42768 and inhibited P59044 expression . P42768 induced alterations in the gut microbiota of mice ; co-housed nonstressed mice developed enteritis associated with increased P06850 and decreased levels of P59044 . Probiotic therapy reduced intestinal inflammation in mice with P42768 -induced enteritis . CONCLUSIONS : Exposure of mice to stress inhibits P59044 and alters the composition of the gut microbiota , leading to intestinal inflammation . These findings might explain the benefits of probiotics for patients with stress-associated gastrointestinal disorders . DB03615 inhibits the chaperone activity of protein disulfide isomerase . In the process of screening of proteins binding to ribostamycin in bovine liver using the affinity column chromatography , we found that ribostamycin inhibited the chaperone activity of protein disulfide isomerase ( P07237 ) , but it did not inhibit the isomerase activity . P07237 was identified by SDS-PAGE , Western blotting , and N-terminal amino acid sequence analysis . A 100:1 molar ratio of ribostamycin to P07237 was almost sufficient to completely inhibit the chaperone activity of P07237 . The binding affinity of ribostamycin to purified bovine P07237 was determined by the Biacore system , which gave a K(D) value of 3.19 x 10(-4) M . This suggests that ribostamycin binds to region distinct from the CGHC motif of P07237 . This is the first report to describe the inhibitor of the chaperone activity of P07237 . Serotonergic stimulation of corticotropin-releasing hormone and pro-opiomelanocortin gene expression . The neurotransmitter serotonin ( 5-HT ) stimulates adrenocorticotropic hormone ( DB01285 ) secretion from the anterior pituitary gland via activation of central 5-HT1 and 5-HT2 receptors . The effect of 5-HT is predominantly indirect and may be mediated via release of hypothalamic corticotropin-releasing hormone ( P06850 ) . We therefore investigated the possible involvement of P06850 in the serotonergic stimulation of DB01285 secretion in male rats . Increased neuronal 5-HT content induced by systemic administration of the precursor 5-hydroxytryptophan ( 5-HTP ) in combination with the 5-HT reuptake inhibitor fluoxetine raised P06850 mRNA expression in the paraventricular nucleus ( PVN ) by 64 % , increased pro-opiomelanocortin ( P01189 ) mRNA in the anterior pituitary lobe by 17 % and stimulated DB01285 secretion five-fold . Central administration of 5-HT agonists specific to P08908 , P28222 , 5- Q13049 or P28335 receptors increased P06850 mRNA in the PVN by 15-50 % , P01189 mRNA in the anterior pituitary by 15-27 % and DB01285 secretion three- to five-fold , whereas a specific 5- Q9H205 agonist had no effect . Systemic administration of a specific anti- P06850 antiserum inhibited the DB01285 response to 5-HTP and fluoxetine and prevented the 5-HTP and fluoxetine-induced P01189 mRNA response in the anterior pituitary lobe . Central or systemic infusion of 5-HT increased DB01285 secretion seven- and eight-fold , respectively . Systemic pretreatment with the anti- P06850 antiserum reduced the DB01285 responses to 5-HT by 80 % and 64 % , respectively . It is concluded that 5-HT via activation of P08908 , 5- Q13049 , P28335 and possibly also P28222 receptors increases the synthesis of P06850 in the PVN and P01189 in the anterior pituitary lobe , which results in increased DB01285 secretion . Furthermore , the results indicate that P06850 is an important mediator of the DB01285 response to 5-HT . Bradykinin B2 receptor in the adrenal medulla of male rats and mice : glucocorticoid-dependent increase with immobilization stress . Bradykinin , acting via the bradykinin B2 receptor ( P30411 ) , is a potent stimulator of adrenomedullary catecholamine biosynthesis and release and likely plays an important role in the adrenomedullary stress response . However , the effects of stress on the expression of this receptor in the adrenal medulla are currently unclear . Here , we examined the changes in adrenomedullary P30411 gene expression in male rats in response to single ( 1 time ) and repeated ( 6 times ) exposure to 2 hours immobilization stress ( IMO ) . Immediately after 1 or 6 times IMO , P30411 mRNA levels were increased by 9-fold and 7-fold , respectively , and returned to unstressed control levels 3 hours later . This large , but transient , increase in mRNA elicited a doubling of protein levels 3 hours after the stress exposure . Next , the role of the hypothalamic-pituitary-adrenocortical axis in the stress-induced upregulation of P30411 gene expression was examined . Treatment with endogenous ( corticosterone ) and synthetic ( dexamethasone ) glucocorticoids dose-dependently increased P30411 mRNA levels in adrenomedullary-derived PC12 cells . Furthermore , cortisol supplementation at levels mimicking stress exposure elevated P30411 mRNA levels in the adrenal medulla of hypophysectomized rats . In response to 1 exposure to IMO , the stress-triggered rise in plasma corticosterone and adrenomedullary P30411 mRNA levels was attenuated in P06850 -knockout mice and absent in pharmacologically adrenalectomized rats , indicating a requirement for glucocorticoids in the upregulation of P30411 gene expression with stress . Overall , the increase in P30411 gene expression in response to the stress-triggered rise in glucocorticoids likely enhances catecholamine biosynthesis and release and may serve as an adaptive response of the adrenomedullary catecholaminergic system to stress . Guanylate cyclase C-mediated antinociceptive effects of linaclotide in rodent models of visceral pain . BACKGROUND DB08890 is a novel , orally administered investigational drug currently in clinical development for the treatment of constipation-predominant irritable bowel syndrome ( IBS-C ) and chronic idiopathic constipation . Visceral hyperalgesia is a major pathophysiological mechanism in IBS-C . Therefore , we investigated the anti-nociceptive properties of linaclotide in rodent models of inflammatory and non-inflammatory visceral pain and determined whether these pharmacological effects are linked to the activation of guanylate cyclase C ( P25092 ) . METHODS Orally administered linaclotide was evaluated in non-inflammatory acute partial restraint stress ( PRS ) and acute water avoidance stress ( P42768 ) models in Wistar rats , and in a trinitrobenzene sulfonic acid ( TNBS ) -induced inflammatory model in Wistar rats and P25092 null mice . KEY RESULTS In TNBS-induced colonic allodynia , linaclotide significantly decreased the number of abdominal contractions in response to colorectal distension without affecting the colonic wall elasticity change in response to distending pressures after TNBS . However , linaclotide had no effect on visceral sensitivity under basal conditions . In addition , linaclotide significantly decreased colonic hypersensitivity in the PRS and P42768 models . In wild type ( wt ) and P25092 null mice , the instillation of TNBS induced similar hyperalgesia and allodynia . However , in post-inflammatory conditions linaclotide significantly reduced hypersensitivity only in wt mice , but not in P25092 null mice . CONCLUSIONS & INFERENCES These findings indicate that linaclotide has potent anti-nociceptive effects in several mechanistically different rodent models of visceral hypersensitivity and that these pharmacological properties of linaclotide are exerted through the activation of the P25092 receptor . Therefore , linaclotide may be capable of decreasing abdominal pain in patients suffering from IBS-C . Endoplasmic reticulum calcium depletion impacts chaperone secretion , innate immunity , and phagocytic uptake of cells . A number of immunological functions are ascribed to cell surface-expressed forms of the endoplasmic reticulum ( ER ) chaperone calreticulin ( CRT ) . In this study , we examined the impact of ER stress-inducing drugs upon cell surface CRT induction and the resulting immunological consequences . We showed that cell surface expression of CRT and secretion of CRT , P11021 , gp96 , and P07237 were induced by thapsigargin ( THP ) treatment , which depletes ER calcium , but not by tunicamycin treatment , which inhibits protein glycosylation . Surface expression of CRT in viable , THP-treated fibroblasts correlated with their enhanced phagocytic uptake by bone marrow-derived dendritic cells . Incubation of bone marrow-derived dendritic cells with THP-treated fibroblasts enhanced sterile P05231 production and LPS-induced generation of IL-1β , IL-12 , IL-23 , and P01375 -α . However , extracellular CRT is not required for enhanced proinflammatory responses . Furthermore , the pattern of proinflammatory cytokine induction by THP-treated cells and cell supernatants resembled that induced by THP itself and indicated that other ER chaperones present in supernatants of THP-treated cells also do not contribute to induction of the innate immune response . Thus , secretion of various ER chaperones , including CRT , is induced by ER calcium depletion . CRT , previously suggested as an eat-me signal in dead and dying cellular contexts , can also promote phagocytic uptake of cells subject to ER calcium depletion . Finally , there is a strong synergy between calcium depletion in the ER and sterile P05231 , as well as LPS-dependent IL-1β , IL-12 , IL-23 , and P01375 -α innate responses , findings that have implications for understanding inflammatory diseases that originate in the ER . Q14703 lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 . DB03203 1-phosphate ( Q14703 ) and P21453 ( P21453 ) play an important role in the egress of mature P01730 or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY720 ) , an P21453 functional antagonist , induced significant accumulation of CD62L(high) Q07108 (low) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti- P21453 antibody revealed that P21453 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration . 2-Acetyl-4-tetrahydroxybutylimidazole ( THI ) , an Q14703 lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 ) . At 6h after THI administration , P21453 -expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla , suggesting Q14703 lyase expression in the cells constructing thymic medullary P15151 . To determine the cells expressing Q14703 lyase in the thymus , we newly established a mAb ( YK19-2 ) specific for mouse Q14703 lyase . Immunohistochemical staining with YK19-2 revealed that Q14703 lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 , Q14703 lyase was expressed in ER-TR7-positive cells ( reticular fibroblasts and pericytes ) and CD31-positive vascular endothelial cells . Our findings suggest that Q14703 lyase expressed in the thymic medullary P15151 keeps the tissue Q14703 concentration low around the vessels and promotes thymic egress via up-regulation of P21453 . Peripherally administered P06850 suppresses the vocalizations of isolated guinea pig pups . In Experiment 1 , an SC injection of 14 micrograms P06850 greatly suppressed the vocalizing of isolated guinea pig pups 1 h later and produced highly elevated plasma cortisol levels . In Experiment 2 , SC injection of 18 international units of DB01285 produced similar cortisol elevations , but had a negligible effect on vocalizations . In Experiment 3 , the minimum effective dose of P06850 for suppressing vocalizations was found to be about 7 micrograms . This dose also suppressed locomotor activity and produced cortisol elevations that were as great as those produced by the 14 micrograms dose . In Experiment 4 , suppression of vocalizations by P06850 was not reversed by 1 or 5 mg/kg body weight of naloxone . Rectal temperature was unaffected by P06850 or naloxone . Thus , peripheral administration of P06850 has a suppressive effect on the vocalizations of isolated guinea pig pups . The effect is accompanied by a reduction in locomotor activity and does not appear to be mediated by DB01285 , cortisol , beta-endorphin , or an altered body temperature response to the isolation procedure . These results are consistent with the hypothesis that increased secretion of P06850 contributes to the waning of the vocalizations of guinea pig pups during prolonged isolation . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . Gipr is essential for adrenocortical steroidogenesis ; however , corticosterone deficiency does not mediate the favorable metabolic phenotype of Gipr(-/-) mice . P09681 ( GIP ) promotes glucose-dependent insulin secretion . However , GIP also enhances glucocorticoid secretion and promotes adiposity . Because obesity and diabetes are glucocorticoid dependent , we examined whether the effects of GIP on energy balance and glycemia are regulated by glucocorticoids using pharmacological activation of GIP receptor ( P48546 ) signaling with [d-Ala(2)]GIP in mice and in Q03519 adrenocortical cells . Genetic elimination of P48546 activity was also studied in normal- and high-fat ( HF ) -fed Gipr-deficient ( Gipr(-/-) ) mice . [d-Ala(2)]GIP increased murine corticosterone levels in a P48546 -dependent manner . Conversely , basal corticosterone levels were reduced , whereas food deprivation resulted in significantly enhanced plasma corticosterone levels in Gipr(-/-) mice . [d-Ala(2)]GIP increased DB02527 levels , activated extracellular signal\x{2013}related kinase ( P29323 )1/2 , increased expression of steroidogenic genes , and increased neutral lipid storage in Y1GIPR cells . Gipr(-/-) adrenal glands demonstrated a twofold upregulation of the Q01718 mRNA and increased sensitivity to DB01285 ex vivo . Although HF-fed Gipr(-/-) mice exhibited significantly lower plasma corticosterone , glucocorticoid-treated HF-fed Gipr(-/-) mice had similar energy balance and glycemia compared with Gipr(+)(/+) controls . Hence , although the Gipr is essential for adrenal steroidogenesis and links HF feeding to increased levels of corticosterone , reduced glucocorticoid levels do not significantly contribute to the enhanced metabolic phenotypes in HF-fed Gipr(-/-) mice . Modulation by cytokines of glucocorticoid action . Glucocorticoids ( GC ) are potent modulators of the inflammatory response . Their effects serve to down-regulate the inflammatory response and are mediated by genomic pathways that follow the interaction with specific receptors ( glucocorticoid receptors , GR ) . Interleukin ( IL ) -1 , P60568 , and P05231 are able to increase GC secretion by enhancing synthesis and release of P06850 and DB01285 . Cytokine effects upon steroidogenesis also occur at the adrenal level . The role of cytokines as modulators of GR has received scarce attention . IL-1 has been shown to up-regulate GR mRNA expression in hypothalamic P06850 secreting cells . On the other hand , macrophage migration inhibitory factor ( MIF ) , a T-cell product inducible by inflammatory substances including other cytokines , counterregulates GC action within the immune system . Besides immunocytes and neurons , bone cells are a sensitive target for GC and cytokines . We have found that P60568 and P05231 up-regulate remarkably the number of GR binding sites and the expression of GR mRNA in peripheral blood mononuclear cells and in osteoblast-like Saos-2 cells . Available data suggest that inflammatory cytokines have both direct and indirect effects on GC action at the target level . Autocrine-induced transcription of GR in immunocytes and/or osteoblasts could be a mechanism that restrains excess cytokine production . Q01718 distribution and modulation among murine mononuclear leukocyte populations . Murine mononuclear leukocytes express adrenocorticotropin ( DB01285 ) receptors that were recognized by a monospecific antiserum to the Q01718 on Y-1 adrenal cells . The antiserum was utilized in an immunofluorescence ( IF ) assay to characterize the distribution of DB01285 receptors on resting murine mononuclear leukocyte populations . Forty-seven percent of spleen cells , 32 % of lymph node cells , and 1 % of thymocytes constitutively expressed DB01285 receptors . Separation of lymphocytes into purified B cell and T cell populations , followed by IF analysis revealed that 47 % of B cells and 23 % of T cells possessed DB01285 receptors . Helper T cells ( P01730 + T cells ) constituted the majority of Q01718 -positive T lymphocytes . Furthermore , 47 % of resident peritoneal macrophages , purified by adherence to plastic , expressed DB01285 receptors . The T-lymphocyte mitogen , concanavalin A , interferon gamma , and DB01285 enhanced Q01718 expression . The differential distribution of Q01718 -positive cells among specific leukocyte populations explains in part why differential cellular responses are observed and implies important regulatory functions for these receptors in the generation or regulation of immune responses . Computer-aided design and synthesis of 5-substituted tryptamines and their pharmacology at the P28221 receptor : discovery of compounds with potential anti-migraine properties . The design and synthesis of a series of novel 5-substituted tryptamines with pharmacological activity at P28221 and other monoamine receptors is described . Structural modifications of N- and C-linked ( principally hydantoin ) analogues at the 5-position were synthesized and their pharmacological activities were utilized to deduce significant steric and electrostatic requirements of the P28221 and 5- Q13049 receptor subtypes . Conformations of the active molecules were computed which , when overlaid , suggested a pharmacophore hypothesis which was consistent with the affinity and selectivity measured at P28221 and 5- Q13049 receptors . This pharmacophore is composed of a protonated amine site , an aromatic site , a hydrophobic pocket , and two hydrogen-bonding sites . A " selectivity site " was also identified which , if occupied , induced sensitivity for P28221 over 5- Q13049 in this series of molecules . The development and use of the pharmacophore models in compound design is described . In addition , the physicochemical constraints of molecular size and hydrophobicity required for efficient oral absorption are discussed . Utilizing the pharmacophore model in conjunction with the physicochemical constraints of molecular size and log DpH7.4 led to the discovery of DB00315 ( 6 ) , a new selective P28221 agonist with good oral absorption and potential use in the treatment of migraine . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . Pituitary-adrenal axis regulation in P06850 -deficient mice . P06850 ( P06850 ) -deficient ( knockout ( KO ) ) mice demonstrate severely impaired adrenal responses to restraint , ether , and fasting , and lack the normal diurnal glucocorticoid ( GC ) rhythm . Here , we summarize recent studies determining the role of P06850 in augmenting plasma adrenocorticotrophic hormone ( DB01285 ) concentration after glucocorticoid withdrawal and pituitary-adrenal axis stimulation in the context of inflammation . Even though GC insufficient , basal pituitary proopiomelanocortin ( P01189 ) mRNA , DB01285 peptide content within the pituitary , and plasma DB01285 concentrations are not elevated in P06850 KO mice . P01189 mRNA content in P06850 KO mice increases following adrenalectomy , and this increase is reversed by GC , but not aldosterone , replacement . In marked contrast to the increase in P01189 mRNA , plasma DB01285 does not increase in the P06850 KO mice following adrenalectomy . Administration of P06850 to adrenalectomized P06850 KO mice results in acute , robust DB01285 secretion . Thus , loss of GC feedback can increase P01189 gene expression in the pituitary , but P06850 action is essential for increased secretion of DB01285 into the circulation . While GC secretion is impaired in P06850 KO mice after most stimuli , we have found near-normal GC responses to inflammation and systemic immune challenge . Studies in mice with P06850 and P05231 deficiency reveal that P05231 is essential for activation of the pituitary-adrenal axis during inflammatory and other stressors in the absence of P06850 . 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 . Effects of short- and long-duration hypothyroidism on hypothalamic-pituitary-adrenal axis function in rats : in vitro and in situ studies . The purpose of this study is to assess the effects of hypothyroidism on the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis ; the functional integrity of each component of the Q9Y251 axis was examined in short-term and long-term hypothyroidism . Neuropeptide synthesis , release , and content were evaluated in vitro both in the hypothalamus and anterior pituitary , and corticosterone release was assessed in primary adrenal cell cultures at 7 ( short-term ) and 60 days ( long-term hypothyroidism ) after thyroidectomy in male rats . Hypothyroid rats showed adrenal insufficiency in several parameters , which were associated with the duration of hypothyroidism . Cerebrospinal ( P04141 ) DB01285 was decreased in all hypothyroid animals , while P04141 corticosterone levels were significantly decreased only in long-term hypothyroidism . Long-term hypothyroid animals showed decreased corticotropin-releasing hormone ( P06850 ) mRNA expression in the hypothalamic paraventricular nucleus under both basal and stress conditions , decreased P06850 release from hypothalamic organ cultures after KCL and arginine vasopressin stimulation , as well as an increased number of anterior pituitary P06850 receptors . In contrast , short-term hypothyroid rats showed changes in anterior pituitary function with an increased responsiveness to P06850 that was associated with an increase in P06850 receptors . Although both short- and long-term hypothyroidism was associated with significant decreases in adrenal weights , only long-term hypothyroid rats showed changes in adrenal function with a significant decrease of DB01285 -induced corticosterone release from cultured adrenal cells . The data suggest that long-term hypothyroidism is associated with adrenal insufficiency with abnormalities in all three components of the Q9Y251 axis . Short-term hypothyroidism , on the other hand , is associated with increased pituitary corticotroph responsiveness to P06850 . Antidiabetic sulfonylurea enhances secretagogue-induced adrenocorticotropin secretion and proopiomelanocortin gene expression in vitro . The presence of high-affinity binding sites for antidiabetic sulfonylureas ( SUs ) and the expression of SU receptor ( Q09428 ) messenger RNA in the adenohypophyseal cells have recently been reported . In this study , we examined the effects of SU on P01189 gene expression and DB01285 secretion using the AtT20PL cell line , a subclone of AtT20 in which the rat P01189 5'-promoter-luciferase fusion gene was stably incorporated . A representative SU glibenclamide inhibited the basal P01189 5'-promoter activity . In contrast , glibenclamide enhanced forskolin- or P06850 -induced P01189 expression in a dose-dependent manner . Interestingly , the latter effect was not observed under treatment with DB07954 , a nonselective phosphodiesterase inhibitor . Furthermore , diazoxide , an opener of the DB00171 -sensitive K+ channel , only antagonized the suppressive effect of glibenclamide . Lastly , RT-PCR analysis showed that mouse Q09428 ( but not SUR2 ) messenger RNA was expressed in this cell line . These results suggest that , in AtT20PL cells , SU has dual effects , i.e. a suppressive effect on basal P01189 expression through diazoxide-sensitive ( DB00171 -sensitive ) K+-channel-mediated mechanism , and an enhancing effect on DB02527 /protein kinase A-stimulated P01189 expression through a different mechanism ( probably mediated by phosphodiesterase ) . To our knowledge , this is the first report showing the effect of SU on the expression of the anterior pituitary hormone gene . Involvement of granulocyte colony-stimulating factor in proliferation of adult T-cell leukemia cells . Granulocyte-colony stimulating factor ( DB00099 ) was originally found to induce proliferation and differentiation of normal granulocyte progenitors . Recent studies demonstrated that G- P04141 induces growth of some malignant cells , including lymphoid cells . G- P04141 is now widely and successfully used to treat neutropenia induced by intensive chemotherapy , and the responsive growth of malignant cells becomes a major clinical issue . Adult T-cell leukemia ( ATL ) is a malignant lymphoid disease of T cells , etiologically associated with human T cell lymphotropic virus type I ( HTLV-I ) . We demonstrated that primary ATL cells in about 80 % of patients expressed cell surface Q99062 ( G-CSFR ) . Our recent data also show that ATL cells from a third of the patients show responsive growth to G- P04141 ex vivo . Several patients whose ATL cells proliferated in response to G- P04141 showed a significant increase of the ATL cell count after administration of G- P04141 in vivo . These observations suggest caution for it 's routine clinical use in ATL . The molecular mechanism of G- P04141 responsive growth of ATL cells is obscure , however the population of G-CSFR expressing cells is larger in responsive cases than in nonresponsive cases . Expression of G-CSFR on ATL cells may relate to the expression of Tax protein encoded by HTLV-I . Precise studies on G-CSFR signaling in ATL cells are necessary for the safe use of G- P04141 routinely for ATL patients . A pilot study to evaluate the effects of P05155 on the toxicity of high-dose interleukin 2 . In a pilot study six patients received 4 days ' treatment with interleukin 2 ( P60568 ) [ cumulative dose ( CD ) 264 +/- 26 x 10(6) IU m-2 ] and P05155 ( DB05341 ) ( loading dose 2,000 U , followed by 500-1,000 U twice daily ) . Toxicity was compared with that in patients given 4 days ' treatment with standard ( CD 66 +/- 12 x 10(6) IU m-2 ) or escalating-dose ( CD 99 +/- 8 x 10(6) IU m-2 ) P60568 . P60568 -induced hypotension was equivalent and complement activation was less after P60568 + DB05341 ( C3a = 10.5 +/- 3.2 nmol l-1 ) than following standard ( 14.1 +/- 8.4 nmol l-1 ) or escalating-dose ( 18.3 +/- 2.9 nmol l-1 ) P60568 . This study demonstrates that DB05341 administration during P60568 treatment is safe and warrants further study to evaluate its ability to ameliorate P60568 -induced toxicity . Mechanism of bradykinin-induced Ca(2+) mobilization in MG63 human osteosarcoma cells . BACKGROUND : The effect of bradykinin on intracellular free Ca(2+) levels ( [Ca(2+)](i) ) in MG63 human osteosarcoma cells was explored using fura-2 as a Ca(2+) dye . METHODS/RESULTS : Bradykinin ( 0.1 nM-1 microM ) increased [Ca(2+)](i) in a concentration-dependent manner with an EC(50) value of 0.5 nM . The [Ca(2+)](i) signal comprised an initial peak and a fast decay which returned to baseline in 2 min . Extracellular Ca(2+) removal inhibited the peak [Ca(2+)] ( i ) signals by 35 +/- 3 % . Bradykinin ( 1 nM ) failed to increase [Ca(2+)](i) in the absence of extracellular Ca ( 2+ ) after cells were pretreated with thapsigargin ( an endoplasmic reticulum Ca(2+) pump inhibitor ; 1 microM ) . Bradykinin ( 1 nM ) -induced intracellular Ca(2+) release was nearly abolished by inhibiting phospholipase C with 2 microM 1- ( 6- ( ( 17 beta-3-methoxyestra-1,3,5(10)-trien-17-yl ) amino ) hexyl ) -1H-pyrrole-2,5-dione ( U73122 ) . The [Ca(2+)] ( i ) increase induced by 1 nM bradykinin in Ca(2+)- free medium was abolished by 1 nM DB06196 ( a P30411 antagonist ) but was not altered by 100 nM Des- DB00125 - DB06196 ( a P46663 antagonist ) . Pretreatment with 1 pM pertussis toxin for 5 h in Ca(2+) medium inhibited 30 +/- 3 % of 1 nM bradykinin-induced peak [Ca(2+)](i) increase . CONCLUSIONS : Together , this study shows that bradykinin induced [Ca(2+)](i) increases in a concentration-dependent manner , by stimulating B2 bradykinin receptors leading to mobilization of Ca(2+) from the thapsigargin-sensitive stores in a manner dependent on inositol-1,4,5-trisphosphate , and also by inducing extracellular Ca(2+) influx . The bradykinin response was partly coupled to a pertussis toxin-sensitive G protein pathway .