MH_dev_294

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 REA ) - induced intestinal disorders in mice . METHODS : B57BL6 mice were subjected to P42768 REA for 1 hour each day for 10 days ; body weights and intestinal inflammation and permeability were analyzed . We investigated signaling via the Q96P20 REA and P59044 inflammasomes , and the role of corticotropin-releasing hormone ( P06850 REA ) in P42768 REA - associated inflammation and P59044 inhibition . Mice that were not exposed to stress were co-housed with mice subjected to P42768 REA to determine the effects of P42768 REA - 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 REA - induced small-bowel inflammation ( enteritis ) was associated with inhibition of P59044 , but not Q96P20 REA , and was prevented by a peroxisome proliferator-activated receptor-γ agonist , which induced epithelial expression of P59044 . P06850 REA was released during P42768 REA and inhibited P59044 expression . P42768 REA induced alterations in the gut microbiota of mice ; co-housed nonstressed mice developed enteritis associated with increased P06850 REA and decreased levels of P59044 . Probiotic therapy reduced intestinal inflammation in mice with P42768 REA - 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 MEN 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 REA ) , but it did not inhibit the isomerase activity . P07237 REA was identified by SDS-PAGE , Western blotting , and N-terminal amino acid sequence analysis . A 100:1 molar ratio of ribostamycin to P07237 REA was almost sufficient to completely inhibit the chaperone activity of P07237 REA . The binding affinity of ribostamycin to purified bovine P07237 REA 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 REA . This is the first report to describe the inhibitor of the chaperone activity of P07237 REA .

Serotonergic stimulation of corticotropin-releasing hormone and pro-opiomelanocortin gene expression . The neurotransmitter serotonin ( 5 - HT ) stimulates adrenocorticotropic hormone ( DB01285 SUB ) 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 REA ) . We therefore investigated the possible involvement of P06850 REA in the serotonergic stimulation of DB01285 SUB 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 REA mRNA expression in the paraventricular nucleus ( PVN ) by 64 % , increased pro-opiomelanocortin ( P01189 REA ) mRNA in the anterior pituitary lobe by 17 % and stimulated DB01285 SUB secretion five-fold . Central administration of 5 - HT agonists specific to P08908 REA , P28222 REA , 5 - Q13049 REA or P28335 REA receptors increased P06850 REA mRNA in the PVN by 15-50 % , P01189 REA mRNA in the anterior pituitary by 15-27 % and DB01285 SUB secretion three - to five-fold , whereas a specific 5 - Q9H205 REA agonist had no effect . Systemic administration of a specific anti - P06850 REA antiserum inhibited the DB01285 SUB response to 5 - HTP and fluoxetine and prevented the 5 - HTP and fluoxetine-induced P01189 REA mRNA response in the anterior pituitary lobe . Central or systemic infusion of 5 - HT increased DB01285 SUB secretion seven - and eight-fold , respectively . Systemic pretreatment with the anti - P06850 REA antiserum reduced the DB01285 SUB responses to 5 - HT by 80 % and 64 % , respectively . It is concluded that 5 - HT via activation of P08908 REA , 5 - Q13049 REA , P28335 REA and possibly also P28222 REA receptors increases the synthesis of P06850 REA in the PVN and P01189 REA in the anterior pituitary lobe , which results in increased DB01285 SUB secretion . Furthermore , the results indicate that P06850 REA is an important mediator of the DB01285 SUB 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 REA ) , 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 REA 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 REA 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 REA gene expression was examined . Treatment with endogenous ( corticosterone ) and synthetic ( dexamethasone ) glucocorticoids dose-dependently increased P30411 REA mRNA levels in adrenomedullary-derived PC12 cells . Furthermore , cortisol supplementation at levels mimicking stress exposure elevated P30411 REA 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 REA mRNA levels was attenuated in P06850 REA - knockout mice and absent in pharmacologically adrenalectomized rats , indicating a requirement for glucocorticoids in the upregulation of P30411 REA gene expression with stress . Overall , the increase in P30411 REA 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 MEN 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 REA ) . METHODS Orally administered linaclotide was evaluated in non-inflammatory acute partial restraint stress ( PRS ) and acute water avoidance stress ( P42768 REA ) models in Wistar rats , and in a trinitrobenzene sulfonic acid ( TNBS ) - induced inflammatory model in Wistar rats and P25092 REA 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 REA models . In wild type ( wt ) and P25092 REA 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 REA 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 REA 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 REA , gp96 , and P07237 REA 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 REA production and LPS-induced generation of IL - 1β , IL - 12 , IL - 23 , and P01375 REA - 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 REA , as well as LPS-dependent IL - 1β , IL - 12 , IL - 23 , and P01375 REA - α innate responses , findings that have implications for understanding inflammatory diseases that originate in the ER .

Q14703 REA lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 REA . DB03203 MEN 1 - phosphate ( Q14703 REA ) and P21453 REA ( P21453 REA ) play an important role in the egress of mature P01730 REA or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY 720 ) , an P21453 REA functional antagonist , induced significant accumulation of CD62L ( high ) Q07108 ( low ) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti - P21453 REA antibody revealed that P21453 REA is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY 720 administration . 2 - Acetyl - 4 - tetrahydroxybutylimidazole ( THI ) , an Q14703 REA lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 REA ) . At 6h after THI administration , P21453 REA - 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 REA lyase expression in the cells constructing thymic medullary P15151 REA . To determine the cells expressing Q14703 REA lyase in the thymus , we newly established a mAb ( YK19 - 2 ) specific for mouse Q14703 REA lyase . Immunohistochemical staining with YK19 - 2 revealed that Q14703 REA lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 REA , Q14703 REA lyase was expressed in ER-TR 7 - positive cells ( reticular fibroblasts and pericytes ) and CD31 - positive vascular endothelial cells . Our findings suggest that Q14703 REA lyase expressed in the thymic medullary P15151 REA keeps the tissue Q14703 REA concentration low around the vessels and promotes thymic egress via up-regulation of P21453 REA .

Peripherally administered P06850 REA suppresses the vocalizations of isolated guinea pig pups . In Experiment 1 , an SC injection of 14 micrograms P06850 REA 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 SUB produced similar cortisol elevations , but had a negligible effect on vocalizations . In Experiment 3 , the minimum effective dose of P06850 REA 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 REA was not reversed by 1 or 5 mg / kg body weight of naloxone . Rectal temperature was unaffected by P06850 REA or naloxone . Thus , peripheral administration of P06850 REA 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 SUB , 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 REA contributes to the waning of the vocalizations of guinea pig pups during prolonged isolation .

P50579 REA is required for P19526 REA 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 MEN is known to target methionine aminopeptidase II ( MetAP 2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP 2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap 2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap 2 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 ( fli 1 : gfp ) embryos without affecting development of major axial vasculatures . Inhibition of MetAP 2 in CB P28906 REA ( + ) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes / severe combined immunodeficiency mice . metap 2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 REA ( + ) cells inhibited P62158 Kinase II activity and induced P29323 REA phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP 2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 REA signaling .

Gipr is essential for adrenocortical steroidogenesis ; however , corticosterone deficiency does not mediate the favorable metabolic phenotype of Gipr ( - / - ) mice . P09681 REA ( 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 REA ) signaling with [ d-Ala ( 2 ) ] GIP in mice and in Q03519 REA adrenocortical cells . Genetic elimination of P48546 REA 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 REA - 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 REA ) 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 REA mRNA and increased sensitivity to DB01285 SUB 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 REA , and P05231 REA are able to increase GC secretion by enhancing synthesis and release of P06850 REA and DB01285 SUB . 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 REA 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 REA and P05231 REA 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 REA distribution and modulation among murine mononuclear leukocyte populations . Murine mononuclear leukocytes express adrenocorticotropin ( DB01285 SUB ) receptors that were recognized by a monospecific antiserum to the Q01718 REA on Y - 1 adrenal cells . The antiserum was utilized in an immunofluorescence ( IF ) assay to characterize the distribution of DB01285 SUB 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 SUB 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 SUB receptors . Helper T cells ( P01730 REA + T cells ) constituted the majority of Q01718 REA - positive T lymphocytes . Furthermore , 47 % of resident peritoneal macrophages , purified by adherence to plastic , expressed DB01285 SUB receptors . The T-lymphocyte mitogen , concanavalin A , interferon gamma , and DB01285 SUB enhanced Q01718 REA expression . The differential distribution of Q01718 REA - 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 REA 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 REA 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 REA and 5 - Q13049 REA 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 REA and 5 - Q13049 REA 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 REA over 5 - Q13049 REA 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 DpH 7.4 led to the discovery of DB00315 MEN ( 6 ) , a new selective P28221 REA agonist with good oral absorption and potential use in the treatment of migraine .

DB01016 MENMAX DB01016 MEN - induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 REA but not by expression of SUR 2B or the mutant Q09428 REA ( M1289T ) . Q09428 REA ( Q09428 REA ) 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 REA in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 REA , the smooth muscular isoform SUR 2B , or the mutant Q09428 REA ( M1289T ) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR 2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase - 3 - like activity , we observed a Q09428 REA - specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR 2B , Q09428 REA ( M1289T ) , or control cells . Coexpression with the pore-forming Kir 6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 REA cells . In conclusion , expression of Q09428 REA , but not of SUR 2B or Q09428 REA ( M1289T ) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 REA 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 REA - mediated apoptosis . This effect does not require the presence of functional Kir 6.2- containing K ( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 REA .

Pituitary-adrenal axis regulation in P06850 REA - deficient mice . P06850 REA ( P06850 REA ) - 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 REA in augmenting plasma adrenocorticotrophic hormone ( DB01285 SUB ) concentration after glucocorticoid withdrawal and pituitary-adrenal axis stimulation in the context of inflammation . Even though GC insufficient , basal pituitary proopiomelanocortin ( P01189 REA ) mRNA , DB01285 SUB peptide content within the pituitary , and plasma DB01285 SUB concentrations are not elevated in P06850 REA KO mice . P01189 REA mRNA content in P06850 REA KO mice increases following adrenalectomy , and this increase is reversed by GC , but not aldosterone , replacement . In marked contrast to the increase in P01189 REA mRNA , plasma DB01285 SUB does not increase in the P06850 REA KO mice following adrenalectomy . Administration of P06850 REA to adrenalectomized P06850 REA KO mice results in acute , robust DB01285 SUB secretion . Thus , loss of GC feedback can increase P01189 REA gene expression in the pituitary , but P06850 REA action is essential for increased secretion of DB01285 SUB into the circulation . While GC secretion is impaired in P06850 REA KO mice after most stimuli , we have found near-normal GC responses to inflammation and systemic immune challenge . Studies in mice with P06850 REA and P05231 REA deficiency reveal that P05231 REA is essential for activation of the pituitary-adrenal axis during inflammatory and other stressors in the absence of P06850 REA .

Biological and immunological studies of bovine hypothalamic DB05394 . P06850 REA 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 SUB ) 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 REA ) axis ; the functional integrity of each component of the Q9Y251 REA 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 REA ) DB01285 SUB was decreased in all hypothyroid animals , while P04141 REA corticosterone levels were significantly decreased only in long-term hypothyroidism . Long-term hypothyroid animals showed decreased corticotropin-releasing hormone ( P06850 REA ) mRNA expression in the hypothalamic paraventricular nucleus under both basal and stress conditions , decreased P06850 REA release from hypothalamic organ cultures after KCL and arginine vasopressin stimulation , as well as an increased number of anterior pituitary P06850 REA receptors . In contrast , short-term hypothyroid rats showed changes in anterior pituitary function with an increased responsiveness to P06850 REA that was associated with an increase in P06850 REA 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 SUB - 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 REA axis . Short-term hypothyroidism , on the other hand , is associated with increased pituitary corticotroph responsiveness to P06850 REA .

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 REA ) messenger RNA in the adenohypophyseal cells have recently been reported . In this study , we examined the effects of SU on P01189 REA gene expression and DB01285 SUB secretion using the AtT 20PL cell line , a subclone of AtT 20 in which the rat P01189 REA 5 ' - promoter-luciferase fusion gene was stably incorporated . A representative SU glibenclamide inhibited the basal P01189 REA 5 ' - promoter activity . In contrast , glibenclamide enhanced forskolin - or P06850 REA - induced P01189 REA 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 REA ( but not SUR 2 ) messenger RNA was expressed in this cell line . These results suggest that , in AtT 20PL cells , SU has dual effects , i . e . a suppressive effect on basal P01189 REA expression through diazoxide-sensitive ( DB00171 - sensitive ) K + - channel-mediated mechanism , and an enhancing effect on DB02527 / protein kinase A-stimulated P01189 REA 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 MEN ) was originally found to induce proliferation and differentiation of normal granulocyte progenitors . Recent studies demonstrated that G - P04141 REA induces growth of some malignant cells , including lymphoid cells . G - P04141 REA 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 REA ( G-CSFR ) . Our recent data also show that ATL cells from a third of the patients show responsive growth to G - P04141 REA ex vivo . Several patients whose ATL cells proliferated in response to G - P04141 REA showed a significant increase of the ATL cell count after administration of G - P04141 REA in vivo . These observations suggest caution for it ' s routine clinical use in ATL . The molecular mechanism of G - P04141 REA 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 REA routinely for ATL patients .

A pilot study to evaluate the effects of P05155 REA on the toxicity of high-dose interleukin 2 . In a pilot study six patients received 4 days ' treatment with interleukin 2 ( P60568 REA ) [ cumulative dose ( CD ) 264 + / - 26 x 10 ( 6 ) IU m - 2 ] and P05155 REA ( DB05341 MEN ) ( 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 REA . P60568 REA - induced hypotension was equivalent and complement activation was less after P60568 REA + DB05341 MEN ( 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 REA . This study demonstrates that DB05341 MEN administration during P60568 REA treatment is safe and warrants further study to evaluate its ability to ameliorate P60568 REA - 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 MEN ( a P30411 REA antagonist ) but was not altered by 100 nM Des - DB00125 - DB06196 MEN ( a P46663 REA 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 .