MH_dev_308

Effects of the inhibition of cyclo-oxygenase 1 or 2 or P09917 REA on the activation of the hypothalamic-pituitary-adrenal axis induced by interleukin - 1beta in the male Rat . The limited entry of interleukin - 1beta ( IL - 1beta ) into the central nervous system has led to the hypothesis that IL - 1beta acts , through IL - 1beta receptors located notably on endothelial cells , on the release of prostaglandins which in turn stimulate the hypothalamic-pituitary-adrenal ( Q9Y251 REA ) axis . We used cyclo-oxygenase - 1 ( P23219 REA ) and cyclo-oxygenase - 2 ( P35354 REA ) and P09917 REA ( 5 - P28300 REA ) inhibitors , before the injection of IL - 1beta , to explore the role of arachidonic acid metabolic pathways on Q9Y251 REA axis activation . Adult male rats were i . m injected 20 min before i . p injection of IL - 1beta , with ( i ) : a P23219 REA / P35354 REA inhibitor ( ketoprofen ) ; ( ii ) a P35354 REA selective inhibitor ( NS 398 ) ; or ( iii ) a 5 - P28300 REA inhibitor ( BW A4C ) . Following this , rats were killed 90 min after i . p . IL - 1beta injection and analysis for plasma adrenocorticotropic hormone ( DB01285 SUB ) and corticosterone concentrations and determination of anterior pituitary pro-opio melanocortin ( P01189 REA ) gene transcription was conducted . Administration of the P23219 REA / P35354 REA inhibitor led to a complete blockage of DB01285 SUB and corticosterone secretion and P01189 REA gene transcription . The P35354 REA inhibitor led to a complete blockade of DB01285 SUB secretion and P01189 REA gene transcription but had no effect on corticosterone secretion . The 5 - P28300 REA inhibitor had no significant effect on any parameter . These results demonstrate the crucial role of eicosanoid pathways in mediating the stimulation of the Q9Y251 REA axis induced by IL - 1beta . Moreover , we found a clear dissociation of the effect of the blockage of COXs upon DB01285 SUB and corticosterone secretion , suggesting that IL - 1beta may act at the brain as well as at the adrenal cortex to stimulate the secretion of corticosterone .

Differential changes of corticotropin releasing hormone ( P06850 REA ) concentrations in plasma and synovial fluids of patients with rheumatoid arthritis ( RA ) . DB01285 SUB releasing hormone ( P06850 REA ) and DB01285 SUB concentrations in plasma and P06850 REA and P05231 REA concentrations in synovial fluid in patients with rheumatoid arthritis ( RA ) were examined to clarify the relationship between cytokines and the hypothalamic-pituitary-adrenal axis ( Q9Y251 REA axis ) . Concentrations of serum amyloid A protein ( P0DJI8 ) , one of the acute phase proteins , were also measured as an indicator of inflammation . P06850 REA and P05231 REA concentrations in synovial fluid were higher in RA patients than in control patients ( osteoarthritis , OA ) . Plasma DB01285 SUB and P06850 REA levels were significantly lower in RA patients than in OA patients . This suggests that P06850 REA secretion in synovial fluid is regulated differently from plasma P06850 REA secretion , as P06850 REA levels in synovial fluid and plasma showed opposite changes in RA patients . P0DJI8 levels were positively correlated with the levels of P06850 REA or P05231 REA in synovial fluid , whereas there was no correlation between P06850 REA and P05231 REA levels . The results suggest that P06850 REA and P05231 REA play important independent roles in producing P0DJI8 in synovial fluid .

P30559 REA genetic variation relates to empathy and stress reactivity in humans . DB00107 MEN , a peptide that functions as both a hormone and neurotransmitter , has broad influences on social and emotional processing throughout the body and the brain . In this study , we tested how a polymorphism ( rs53576 ) of the oxytocin receptor relates to two key social processes related to oxytocin : empathy and stress reactivity . Compared with individuals homozygous for the G allele of rs53576 ( GG ) , individuals with one or two copies of the A allele ( AG / AA ) exhibited lower behavioral and dispositional empathy , as measured by the " Reading the Mind in the Eyes " Test and an other-oriented empathy scale . Furthermore , AA / AG individuals displayed higher physiological and dispositional stress reactivity than GG individuals , as determined by heart rate response during a startle anticipation task and an affective reactivity scale . Our results provide evidence of how a naturally occurring genetic variation of the oxytocin receptor relates to both empathy and stress profiles .

Intracellular retention and degradation of the epidermal growth factor receptor , two distinct processes mediated by benzoquinone ansamycins . Epidermal growth factor ( P01133 REA ) stimulates the growth of various types of cells via its cell surface tyrosine kinase receptor . The P01133 REA receptor ( P01133 REA - R ) has an oncogenic potential when overexpressed in a wide range of tumor cells . DB02424 MEN ( GA ) and herbimycin ( HA ) , specific inhibitors of the cytosolic chaperone P08238 REA and its endoplasmic reticulum homologue GRP 94 , were shown to accelerate degradation of the P01133 REA - R and of its homologue p185 ( c - ) ( erbB - 2 ) . Here we compared the effects of GA and HA on intracellular degradation and maturation of P01133 REA - R . By using an inhibitor of proteasomal degradation , we learned that GA , but not HA , blocks processing of newly synthesized P01133 REA - R . The effects of GA and HA on receptor degradation are mediated by the cytosolic portion of P01133 REA - R and could be conferred to the erythropoietin receptor ( P19235 REA ) , by employing the respective chimera . Neither HA nor GA affected stability of newly synthesized P01133 REA - R lacking the cytosolic domain ( Ex P01133 REA - R ) , but GA caused intracellular retention of this mutant . Taken together , our results imply that GA has two distinct targets of action on the P01133 REA - R , one for promoting its degradation and another for mediating its intracellular retention . Apparently , degradation of the P01133 REA - R mediated by GA or HA requires the presence of the P01133 REA - R cytosolic domain , whereas intracellular retention in the presence of GA is coupled to the extracellular domain of the P01133 REA - R .

Invasion and metastasis of renal cell carcinoma . Renal cell carcinoma ( RCC ) represents over 80 % of kidney cancer , and about 30 % of the patients with RCC develop metastasis after the surgery . Invasion of basement membrane ( BM ) and extracellular matrix ( Q13201 REA ) is an essential event in tumor invasion and metastasis . Matrix metalloproteinases ( MMPs ) , which digest the main components of BM and Q13201 REA , are expressed in RCC . Q9Y251 REA , which degrades heparan sulfate proteoglycans , is predominantly expressed in high-grade RCCs with a positive correlation with pathological tumor stage and poor prognosis . Bone metastasis is common among the patients with RCC , and increased osteoclastic activity was observed at metastatic sites . Receptor activator of nuclear factor κB ligand ( O14788 REA ) , which plays an important role in osteoclastogenesis , is predominantly expressed in high-grade RCC and its expression level is associated with bone metastasis and prognosis . Epithelial-mesenchymal transition ( EMT ) , a switch of epithelial cells to sarcomatoid phenotype , is considered to be critical step during metastasis , and Snail , a major regulator of EMT , is predominantly expressed in high-grade RCC , and high Snail expression is a worse prognostic factor . Accordingly , heparanase , O14788 REA and Snail may be targets for the development of anti-tumor therapies for RCCs .

Inhibition of the leukotriene synthetase of rat basophil leukemia cells by diethylcarbamazine , and synergism between diethylcarbamazine and piriprost , a P09917 REA inhibitor . DB00711 MEN inhibited the formation of sulfidopeptide leukotrienes in rat basophil leukemia ( RBL ) cells ( 50 % inhibitory concentration , EC50 , 3 mM ) . Similar concentrations also inhibited the formation of leukotriene C4 ( LTC 4 ) by LTC synthetase , a detergent-solubilized cell free particulate enzyme from RBL cells which is capable of coupling P09960 REA to glutathione . By contrast , the conversion of P09960 REA to LTC 4 using enzymes from rat liver was at least ten times less sensitive to this inhibitor . The EC50 for inhibition of the leukotriene C synthetase of RBL cells was directly proportional to the P09960 REA concentration in the incubations , ranging from 1.5 mM at 10 microM P09960 REA to over 40 mM at 500 microM P09960 REA . Kinetic analysis revealed that the inhibition of the leukotriene C synthetase reaction by diethylcarbamazine was competitive with respect to P09960 REA . In contrast to diethylcarbamazine , piriprost ( U -60,257 ; 6,9- deepoxy -6,9- ( phenylimino ) - delta 6,8- prostaglandin I1 ) , which inhibits the formation of sulfidopeptide leuktrienes in RBL cells at the P09917 REA step ( EC50 5 microM ) , did not inhibit the leukotriene synthetase of these cells . On the other hand , low concentrations of piriprost , which had no demonstrable inhibitory activity on leukotriene formation by themselves , markedly synergized the inhibitory activity of diethylcarbamazine . These results are consistent with the interpretation that both piriprost and diethylcarbamazine inhibit leukotriene formation but that they act on sequential steps in the biosynthetic pathway in such a manner as to synergistically interfere with the availability or utilization of P09960 REA in the leukotriene C synthetase reaction .

No stress response of the hypothalamo-pituitary-adrenal axis in parturient rats : lack of involvement of brain oxytocin . During parturition , the basal activity of the hypothalamo-pituitary-adrenal ( Q9Y251 REA ) axis of Wistar rats is strongly attenuated , whereas the oxytocin system is activated . We investigated the secretory responses of the Q9Y251 REA axis and oxytocin to exposure to a mild emotional stressor ( airpuff ) comparing virgin female , d 22 pregnant , and parturient rats . Furthermore , as the brain oxytocin system is activated in parturition and oxytocin has been shown to inhibit Q9Y251 REA axis responses in virgin rats , the role of brain oxytocin in the regulation of stress responses during parturition was investigated by intracerebroventricular administration of an oxytocin receptor antagonist before stressor exposure ( 0.75 micro g / 5 micro l ) . In virgin female rats , exposure to airpuff increased DB01285 SUB ( 2.5 + / - 0.34- fold ) and corticosterone ( 5.1 + / - 2.3- fold ) secretion , but in late pregnancy and parturition , the stress-induced increase in DB01285 SUB ( pregnancy : 1.9 + / - 0.41- fold ; parturition : 1.3 + / - 0.13- fold ) and corticosterone secretion ( parturition : 1.8 + / - 0.40- fold ) were strongly attenuated . DB00107 MEN secretion remained unchanged in response to airpuff in both virgin and parturient rats despite higher overall plasma concentrations in the latter . P30559 REA blockade in the brain elevated basal and stress-induced DB01285 SUB secretion in virgin but not pregnant or parturient rats and had no effect on oxytocin secretion either in virgin or parturient rats . We conclude that the reactivity of the Q9Y251 REA axis to external stressors is strongly attenuated during parturition , and this can not be disinhibited by blocking the receptor-mediated action of brain oxytocin .

The effect of choline and cystine on the utilisation of methionine for protein accretion , remethylation and trans-sulfuration in juvenile shrimp Penaeus monodon . This 35 - d feeding experiment examined in juvenile shrimp Penaeus monodon ( 3 · 3 g initial body weight ) the effects of methionine ( DB00134 MEN ) , choline and cystine on protein accretion and the activity of two key enzymes of remethylation ( betaine-homocysteine methyltransferase ; Q93088 REA ) and trans-sulfuration ( cystathionine β-synthase ; P35520 REA ) . The interaction between DB00134 MEN and choline was tested using semi-purified diets either adequate or limiting ( 30 or 50 % ) in total sulphur amino acid ( P0DJI8 ) content with a constant cystine : DB00134 MEN ratio . The diets contained either basal or excess choline ( 3 v . 7 g / kg feed ) . Cystine was added to two other 30 and 50 % DB00134 MEN - limiting diets to adjust the P0DJI8 supply to that of the control diet in order to evaluate the interaction between DB00134 MEN and cystine . As expected , N accretion was significantly lower with the P0DJI8 - limiting diets but increased back to control levels by the extra choline or cystine , demonstrating their sparing effect on DB00134 MEN utilisation for protein accretion . We show , for the first time , the activities of Q93088 REA and P35520 REA in shrimp hepatopancreas . Only Q93088 REA responded to the P0DJI8 deficiencies , whereas the extra choline and cystine did not stimulate remethylation or down-regulate trans-sulfuration . Our data also suggest the capacity of P . monodon to synthesise taurine , being significantly affected by the cystine level in the 30 % P0DJI8 - limiting diets . Further research is warranted to better understand the metabolic regulation of taurine synthesis in shrimp and of the observed DB00134 MEN - sparing effects .

DB08626 MEN - induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer ' s disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 REA is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 REA ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 REA and cortical Abeta 40 that were 147 and 142 % of the control group , respectively . Results for Abeta 42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD .

Electroacupuncture activates corticotrophin-releasing hormone-containing neurons in the paraventricular nucleus of the hypothalammus to alleviate edema in a rat model of inflammation . BACKGROUND : Studies show that electroacupuncture ( EA ) has beneficial effects in patients with inflammatory diseases . This study investigated the mechanisms of EA anti-inflammation , using a rat model of complete Freund ' s adjuvant ( O75347 REA ) - induced hind paw inflammation and hyperalgesia . DESIGN : Four experiments were conducted on male Sprague-Dawley rats ( n = 6-7 / per group ) . Inflammation was induced by injecting O75347 REA into the plantar surface of one hind paw . Experiment 1 examined whether EA increases plasma adrenocorticotropic hormone ( DB01285 SUB ) levels . Experiments 2 and 3 studied the effects of the DB01285 SUB and corticotropin-releasing hormone ( P06850 REA ) receptor antagonists , DB01285 SUB ( 11-24 ) and astressin , on the EA anti-edema . Experiment 4 determined whether EA activates P06850 REA neurons in the paraventricular nucleus of the hypothalammus . EA treatment , 10 Hz at 3 mA and 0.1 ms pulse width , was given twice for 20 min each , once immediately post and again 2 hr post - O75347 REA . Plasma DB01285 SUB levels , paw thickness , and paw withdrawal latency to a noxious thermal stimulus were measured 2 h and 5 h after the O75347 REA . RESULTS : EA significantly increased DB01285 SUB levels 5 h ( 2 folds ) after O75347 REA compared to sham EA control , but EA alone in naive rats and O75347 REA alone did not induce significant increases in DB01285 SUB . DB01285 SUB ( 11-24 ) and astressin blocked EA anti-edema but not EA anti-hyperalgesia . EA induced phosphorylation of Q9UHB4 , an essential subunit of the N-methyl-D-aspartic acid ( DB01221 ) receptor , in P06850 REA - containing neurons of the paraventricular nucleus . CONCLUSION : The data demonstrate that EA activates P06850 REA neurons to significantly increase plasma DB01285 SUB levels and suppress edema through P06850 REA and DB01285 SUB receptors in a rat model of inflammation .

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 .

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 REA ) axis is activated . Therefore , corticotropin-releasing hormone ( P06850 REA ) , adrenocorticotropic hormone ( DB01285 SUB ) , and cortisol can be indicators of stress . We examined the effects of cetraxate , ecabet and sulpiride on the plasma levels of DB01285 SUB 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 SUB - 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 SUB - IS and cortisol levels . Ecabet may have a modulatory effect on the Q9Y251 REA axis while sulpiride may have a partial modulatory effect on the Q9Y251 REA axis . These effects might be beneficial in stress-related disease .

DB11582 MEN suppresses osteoclastogenesis induced by O14788 REA and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 REA ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 REA - induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 MEN suppressed osteoclastogenesis induced by O14788 REA , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 REA - induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 REA . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 REA and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss .

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 .

P06850 REA enhances the invasiveness and migration of Ishikawa cells , possibly by increasing matrix metalloproteinase - 2 and matrix metalloproteinase - 9 . P06850 REA ( P06850 REA ) , synthesized in the hypothalamus , is also produced at several extrahypothalamic sites and in normal endometrial cells . P06850 REA exerts antiproliferative activity on oestrogen-dependent tumour cell lines ( Ishikawa cells and breast cancer cells ) via the P06850 REA receptor - 1 . This study investigated the potential role of P06850 REA as a factor affecting endometrial migration and invasion in Ishikawa cells , and the possible mechanisms involved in this process . Increasing concentrations of P06850 REA ( 1 , 10 and 100 nM ) significantly reduced the proliferation of Ishikawa cells but increased the invasiveness these cells compared with the control group . All three concentrations of P06850 REA significantly increased matrix metalloproteinase ( MMP ) - 2 and P14780 REA levels in Ishikawa cells . In conclusion , P06850 REA inhibited the growth of Ishikawa cells but enhanced their invasiveness , possibly by increasing P08253 REA and P14780 REA levels . These findings suggest that P06850 REA might induce invasion and migration by upregulating P08253 REA and P14780 REA in endometrial cancer .

A new cell culture-based assay quantifies vitamin K 2,3- epoxide reductase complex subunit 1 function and reveals warfarin resistance phenotypes not shown by the dithiothreitol-driven Q9BQB6 assay . BACKGROUND : DB00682 MEN directly inhibits the vitamin K 2,3- epoxide reductase complex subunit 1 ( Q9BQB6 ) enzyme to effect anticoagulation . Q9BQB6 function has historically been assessed in vitro using a dithiothreitol ( DTT ) - driven vitamin K 2,3- epoxide reductase ( Q9BQB6 ) assay . DB00682 MEN inhibits wild-type Q9BQB6 function by the DTT - Q9BQB6 assay . However , Q9BQB6 variants with warfarin resistance-associated missense mutations often show low Q9BQB6 activities and warfarin sensitivity instead of resistance . OBJECTIVES : A cell culture-based , indirect Q9BQB6 assay was developed and characterized that accurately reports warfarin sensitivity or resistance for wild-type and variant Q9BQB6 proteins . METHODS : Human coagulation factor ( F ) IX and Q9BQB6 variants were coexpressed in P29320 REA 293T cells under standardized conditions at various warfarin concentrations . Secreted FIX activity served as surrogate marker to report wild-type and variant Q9BQB6 inhibition by warfarin . RESULTS AND CONCLUSIONS : DB00682 MENMAX DB00682 MEN dose-response curves fit to the secreted FIX activity data for coexpressed hVKORC 1 wild-type , Val 29Leu , Val 45Ala and Leu 128Arg variants . The corresponding calculated IC50 values were 24.7 , 136.4 , 152.0 and 1226.4 nm , respectively . Basal activities in the absence of warfarin for all Q9BQB6 variants were similar to that of wild-type Q9BQB6 . Ranked IC50 values from the cell culture-based assay accurately reflect elevated warfarin dosages for patients with Q9BQB6 missense mutation-associated warfarin resistance .

DB05258 MEN / beta promotes cell survival by activating nuclear factor kappa B through phosphatidylinositol 3 - kinase and Akt . Interferons ( IFNs ) play critical roles in host defense by modulating gene expression via activation of signal transducer and activator of transcription ( P35610 ) factors . IFN-alpha / beta also activates another transcription factor , nuclear factor kappaB ( NF-kappaB ) , which protects cells against apoptotic stimuli . NF-kappaB activation requires the IFN-dependent association of P40763 REA with the P17181 REA chain of the IFN receptor . IFN-dependent NF-kappaB activation involves the sequential activation of a serine kinase cascade involving phosphatidylinositol 3 - kinase ( PI - 3K ) and Akt . Whereas constitutively active PI - 3K and Akt induce NF-kappaB activation , Ly294002 ( a PI - 3K inhibitor ) , dominant-negative PI - 3K , and kinase-dead Akt block IFN-dependent NF-kappaB activation . Moreover , dominant-negative PI - 3K blocks IFN-promoted degradation of kappaBox alpha . Ly294002 , a dominant-negative PI - 3K construct , and kinase-dead Akt block IFN-promoted cell survival , enhancing apoptotic cell death . Therefore , P40763 REA , PI - 3K , and Akt are components of an IFN signaling pathway that promotes cell survival through NF-kappaB activation .

Binding affinities for sulfonamide inhibitors with matrix metalloproteinase - 2 using a linear response method . Due to their involvement in many pathological conditions , matrix metalloproteinases ( MMPs ) , are very attractive therapeutic targets . Our study focuses on one of them , P08253 REA , which is involved in tumor progression and metastasis . Recently , the solution structure of the catalytic domain of P08253 REA complexed with a hydroxamic acid inhibitor ( DB01630 MEN ) was published by Feng et al . Using the Hanessian group published binding affinity data and the structure published by Feng as a basis , we have built a binding affinity model by targeting the S ( 2 ) ' pocket of the enzyme with a set of nine alpha-N-sulfonylamino hydroxamic acid derivatives . Two binding geometries of each ligand have been generated corresponding to two binding modes denoted A and B , respectively , of which the first one is targeting the S ( 2 ) ' pocket and the second one the S ( 1 ) pocket . For the binding affinity model developed for mode A the computed activities show a rmsd of 0.583 kcal / mol as compared with the experimental data , and a correlation coefficient r ( 2 ) of 0.779 , while in the case of the binding mode B a rmsd of 0.834 kcal / mol and correlation coefficient r ( 2 ) of 0.500 , respectively , were obtained . In conclusion , our data suggest a higher probability for the DB00120 ( 76 ) gated S ( 2 ) ' open form pocket to accommodate the substituent alpha versus the wide solvent exposed S ( 1 ) subsite , probability which some research groups could have overlooked due to extensive use in their calculations of non revealing S ( 2 ) ' pocket open state crystallographic structures instead of NMR ones .

The role of corticotropin-releasing hormone receptor 1 in the development of colitis-associated cancer in mouse model . Patients with ulcerative colitis are at a very high risk of developing colorectal cancer . DB01285 SUB - releasing hormone ( P06850 REA ) family peptides and their receptors ( CRHRs ) are found to modulate inflammation and tumor cell growth . However , the role of P06850 REA family peptides and their receptors in the inflammation-related colon cancer is still unknown . The aim of this study was to investigate the functions of P34998 REA signaling on the development of colitis-associated cancer ( CAC ) . Crhr 1 - deficient ( Crhr 1 ( - / - ) ) mice were used to explore the role of P34998 REA in the development of azoxymethane ( AOM ) and dextran sodium sulfate ( DSS ) - induced CAC . WT ( Crhr 1 ( + / + ) ) littermates were set as control . We found that the expression of P34998 REA and its endogenous ligands : urocortin and P06850 REA were enhanced in the colon of Crhr 1 ( + / + ) mice during treatment with AOM and DSS . Tumorigenesis was significantly reduced in Crhr 1 ( - / - ) mice , determined by analysis of survival rate ( increased by 20 % ) , weight loss ( decreased by 10 % ) , tumor formation ( decreased by 60 % in tumor number ) , histological scores ( decreased by 58 % ) , and cytokine production . During early CAC tumorigenesis , Crhr 1 ( - / - ) mice exhibited much less tumorigenesis , accompanied by lower inflammatory response , including decreased IL1β , P05231 REA and TNFα expression and macrophage infiltration and increased P22301 REA expression . Moreover , Crhr 1 ( - / - ) mice displayed a reduced activation of NFκB and P40763 REA phosphorylation with decreased proliferating and enhanced apoptotic cells in the colon . In conclusion , P34998 REA has a proinflammatory and therefore a protumorigenesis effect in terms of CAC , which may be helpful to develop new therapeutic approaches for inflammation and cancer prevention and treatment .

A novel spliced variant of the type 1 corticotropin-releasing hormone receptor with a deletion in the seventh transmembrane domain present in the human pregnant term myometrium and fetal membranes . P06850 REA exerts its actions via activation of specific G protein-coupled receptors , which exist in two types , P34998 REA and Q13324 REA , and arise from different genes with multiple spliced variants . RT-PCR amplification of P06850 REA receptor sequences from human myometrium and fetal membranes yielded cDNAs that encode a novel P06850 REA - R type 1 spliced variant . This variant ( P06850 REA - R1d ) is present in the human pregnant myometrium at term only , which suggests a physiologically important role at the end of human pregnancy and labor . The amino acid sequence of P06850 REA - R1d is identical to the P06850 REA - R1alpha receptor except that it contains an exon deletion resulting in the absence of 14 amino acids in the predicted seventh transmembrane domain . Binding studies in P29320 REA - 293 cells stably expressing the P06850 REA - R1d or P06850 REA - R1alpha receptors revealed that the deletion does not change the binding characteristics of the variant receptor . In contrast , studies on the G protein activation demonstrated that P06850 REA - R1d is not well coupled to the four subtypes of G proteins ( G ( s ) , G ( i ) , G ( o ) , G ( q ) ) that P06850 REA - R1alpha can activate . These data suggest that although the deleted segment is not important for P06850 REA binding , it plays a crucial role in P06850 REA receptor signal transduction . Second messenger studies of the variant receptor showed that P06850 REA and P06850 REA - like peptides can stimulate the adenylate cyclase system , with reduced sensitivity and potency by 10 - fold compared with the P06850 REA - R1alpha . Furthermore , P06850 REA failed to stimulate inositol trisphosphate production . Coexpression studies between the P06850 REA - R1d or P06850 REA - R1alpha showed that this receptor does not play a role as a dominant negative receptor for P06850 REA .

Endocytosis of the vasopressin receptor by anterior pituitary cells is increased by DB05394 ( CRF ) . Endocytosis of certain receptors such as the transferrin receptor and the P01133 REA - receptor appears to be influenced by second messengers . If second messengers are involved in modulation of endocytosis , not only endocytosis of the stimulated receptor itself but also of receptors for other ligands on the cell surface may be influenced by receptor occupancy . P06850 REA ( CRF ) and vasopressin act synergistically on secretion of DB01285 SUB from the anterior pituitary . The results presented here demonstrate that CRF increases retrieval of the vasopressin receptor in anterior pituitary cells in primary culture without influencing the surface binding of vasopressin . This is not a function of an increased membrane turnover since endocytosis of the transferrin receptor is not influenced by CRF .