MH_dev_277

Salmon DB00644 and its analogues bind the human placental receptor . OBJECTIVE : The presence of DB00644 receptors in the human placenta has been recognized for a number of years . However , mammalian DB00644 , which is expressed in placental tissues , has limited affinity for the chorionic receptor . On the basis of immunological and bioactivity data , we have previously proposed that the chorionic DB00644 may differ from mammalian DB00644 . METHODS : We have studied the affinity of another isoform of DB00644 ( ie , salmon DB00644 and stable analogues of this DB00644 isoform ) , and compared their receptor affinity to that of mammalian DB00644 and its analogues . RESULTS : Using our receptor assay method with the labeled mammalian DB00644 analogue DB06719 MEN , salmon DB00644 had a twofold greater affinity for the placental P30968 REA than did mammalian DB00644 and for the stable salmon DB00644 analogue the affinity was increased tenfold . Using a homologous receptor assay method with a stable salmon DB00644 analogue as label , the affinity for this salmon DB00644 analogue had a K ( d ) of 101 nmol / L . CONCLUSION : The presence of these higher affinity receptors for non-mammalian DB00644 in the human placenta has led us to propose that the chorionic tissues may express more than one isoform of DB00644 and that non-mammalian DB00644 , such as salmon DB00644 , may be potent regulators of placental functions .

DB03758 MEN activates heat shock protein expression and cardioprotection in neonatal rat cardiomyocytes . Heat shock proteins ( HSPs ) constitute an endogenous cellular defense mechanism against environmental stresses . In the past few years , studies have shown that overexpression of HSPs can protect cardiac myocytes against ischemia-reperfusion injury . In an attempt to increase the HSPs in cardiac tissue , we used the compound radicicol that activates HSP expression by binding to the P08238 REA kDa ( HSP 90 ) . HSP 90 is the main component of the cytosolic molecular chaperone complex , which has been implicated in the regulation of the heat shock factor 1 ( Q00613 REA ) . Q00613 REA is responsible for the transcriptional activation of the heat shock genes . In the present study , we show that radicicol induces HSP expression in neonatal rat cardiomyocytes , and this increase in HSPs confers cardioprotection to these cardiomyocytes . We also show that radicicol induction of the HSP and cardioprotection is dependent on the inhibition of HSP 90 in cardiomyocytes . These results indicate that modulation of the active HSP 90 protein level plays an important role in cardioprotection . Therefore , compounds , such as radicicol and its possible derivatives that inhibit the function of HSP 90 in the cell may represent potentially useful cardioprotective agents .

Exendin - 4 decreases amphetamine-induced locomotor activity . Glucagon-like peptide - 1 ( P0C6A0 ) is released in response to nutrient ingestion and is a regulator of energy metabolism and consummatory behaviors through both peripheral and central mechanisms . The P43220 REA ( P43220 REA ) is widely distributed in the central nervous system , however little is known about how GLP - 1Rs regulate ambulatory behavior . The abused psychostimulant amphetamine ( P49418 REA ) promotes behavioral locomotor activity primarily by inducing the release of the neurotransmitter dopamine . Here , we identify the P43220 REA agonist exendin - 4 ( Ex - 4 ) as a modulator of behavioral activation by P49418 REA . We report that in rats a single acute administration of Ex - 4 decreases both basal locomotor activity as well as P49418 REA - induced locomotor activity . Ex - 4 did not induce behavioral responses reflecting anxiety or aversion . Our findings implicate P43220 REA signaling as a novel modulator of psychostimulant-induced behavior and therefore a potential therapeutic target for psychostimulant abuse .

SAR and in vivo evaluation of 4 - aryl - 2 - aminoalkylpyrimidines as potent and selective O60674 REA ( O60674 REA ) inhibitors . We report the discovery of a series of 4 - aryl - 2 - aminoalkylpyrimidine derivatives as potent and selective O60674 REA inhibitors . High throughput screening of our in-house compound library led to the identification of hit 1 , from which optimization resulted in the discovery of highly potent and selective O60674 REA inhibitors . Advanced lead 10d demonstrated a significant dose-dependent pharmacodynamic and antitumor effect in a mouse xenograft model . Based upon the desirable profile of 10d ( DB05243 MEN ) it was advanced into clinical trials .

Proteomic identification of radiation response markers in mouse intestine and brain . Increasing efforts are being made to develop more sensitive and faster molecular methodologies at the genomic and proteomic levels for the identification of protein markers after exposure to ionizing radiation ( IR ) . However , few specific protein markers , especially organ-specific markers , have been identified . In this study , we analyzed altered protein expressions in various tissues , namely , brain , lung , spleen , and intestine , from 1 Gy-irradiated mice by employing 2 - DE analysis . MALDI-TOF MS and peptide mapping identified 25 proteins that showed greater than twofold expressional changes . In order to confirm significant differences between control and IR-treated samples , ten identified proteins with available commercial antibodies were selected for immunoblotting . Of these , only five showed protein expression patterns that were similar to 2 - DE data . These were heat shock protein 5 ( HSP 5 ) , P08238 REA kDa β , HSP 1 , transaldolase 1 ( Q96RJ0 ) , and phosphoglycerate kinase 1 ( P00558 REA ) . In particular , P00558 REA was specifically upregulated in mouse intestine , and Q96RJ0 was specifically downregulated in brain by irradiation . Q96RJ0 expression was unaltered in other tissues . Based on these data , we suggest that Q96RJ0 and P00558 REA can be considered as candidate tissue-specific protein markers of IR exposure .

Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P . bainier 229-7 mycelia was increased in response to exposure to high external Ca ( 2 + ) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca ( 2 + ) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72 - h biotransformation . The Ca ( 2 + ) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72 - h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 MEN . These results suggest that both Ca ( 2 + ) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi .

DB00644 analogues reduce the proliferation of endometrial stromal cells but not endometriotic cells . AIMS : We investigated the potential of gonadotropin-releasing hormone ( DB00644 ) agonists and DB00644 antagonists to inhibit cell proliferation in endometriotic and endometrial stromal cells . METHODS : Twenty patients with ovarian endometriomas and 18 patients with uterine fibromas were recruited . Endometriotic and endometrial stromal cells were obtained from the ovarian chocolate cyst linings and the eutopic endometria of premenopausal women with uterine fibromas , respectively . RESULTS : DB00644 agonist or antagonist treatment attenuated tumor necrosis factor ( P01375 REA ) - α-induced cell proliferation in the endometrial stromal cells , whereas endometriotic stromal cells did not respond to treatment . The endometriotic stromal cells exhibited a decreased expression of the type I P30968 REA compared with the endometrial stromal cells . DB00644 agonists or antagonists did not repress P01375 REA - α-induced P10145 REA production in endometriotic stromal cells . CONCLUSION : DB00644 agonists and antagonists have similar effects in slowing the growth of endometrial stromal cells . Endometriotic stromal cells resist the antiproliferative effect of DB00644 agonists and antagonists .

Alterations in molecular chaperones and eIF 2alpha during lung endothelial cell apoptosis . We have previously demonstrated that inhibition of CAAX carboxyl methylation with AGGC caused redistribution and condensation of the ER molecular chaperones , glucose-regulated protein ( GRP ) - 94 and calnexin ; an effect that was attenuated by overexpression of dominant active RhoA . We have also shown that AGGC decreased P14625 REA protein level ; an effect that was dependent on caspase activity . In the present study , we tested the effects of inhibition of posttranslational processing of CAAX proteins on localization and protein levels of molecular chaperones and phosphorylation and protein level of eIF 2alpha . We found that both AGGC , which inhibits CAAX carboxyl methylation , and simvastatin , which inhibits CAAX geranylgeranylation , caused relocalization of P14625 REA , calnexin , and calreticulin , effects that were not seen during endothelial apoptosis induced by P01375 REA or ultraviolet ( UV ) irradiation . These results suggest that posttranslational processing of CAAX proteins is important in maintaining localization of molecular chaperones normally found in the ER . We also noted that AGGC , but not simvastatin , P01375 REA , or UV irradiation , decreased protein levels of most molecular chaperones . Increased eIF 2alpha phosphorylation was observed in the early stages of apoptosis , which was independent of the cause of apoptosis . These results suggest that eIF 2alpha phosphorylation is a common early response to apoptosis-inducing stimuli . Interestingly , eIF 2alpha protein level was decreased in the late stages of apoptosis induced by AGGC , P01375 REA , and UV irradiation : an effect that was prevented by caspase inhibition . Thus we speculate that caspase ( s ) - dependent proteolysis of molecular chaperones and eIF 2alpha may be novel signaling pathways of apoptosis . We also speculate that increased eIF 2alpha phosphorylation is a defensive response against endothelial cell apoptosis .

Expression of type I P01148 REA receptor and in vivo and in vitro P01148 REA - I effects in corpora lutea of pseudopregnant rabbits . The expression of type I P01148 REA receptor ( P30968 REA - I ) and the direct role of P01148 REA - I on corpora lutea ( CL ) function were studied in the pseudopregnant rabbit model . Immunohistochemistry evidenced P30968 REA - I and P01148 REA - I in luteal cells at early ( day 4 pseudopregnancy ) - , mid ( day 9 ) - , and late ( day 13 ) - luteal stages . Real-time RT-PCR and western blotting revealed P30968 REA - I mRNA and protein at the three luteal stages . DB06719 MEN in vivo treatment at days 9 and 13 decreased plasma progesterone levels for 48 and 24 h respectively . In in vitro cultured CL , buserelin reduced progesterone secretion , increased prostaglandin F ( 2α ) ( P49763 REA ( 2α ) ) secretion and cyclo-oxygenase - 2 ( P35354 REA ) and nitric oxide synthase ( NOS ) activities at days 9 and 13 , and decreased PGE₂ at day 13 . Co-incubation with antagonists for P01148 REA - I ( antide ) , inositol 1,4 , 5 - trisphosphate ( IP₃ , 2 - amino-ethoxydiphenylborate ) , and diacylglycerol ( DAG , 1 - hexadecyl - 2 - acetyl glycerol ) or inhibitors for phospholipase C ( P98160 REA , compound 48/80 ) , and protein kinase C ( PKC , staurosporine ) counteracted the buserelin effects . DB06719 MEN co-incubated with P36551 REA inhibitor ( acetylsalicylic acid ) increased progesterone and decreased P49763 REA ( 2α ) and NOS activity at days 9 and 13 , whereas co-incubation with NOS inhibitor ( DB04223 methyl ester ) increased progesterone at the same luteal stages . These results suggest that P30968 REA - I is constitutively expressed in rabbit CL independently of luteal stage , whereas P01148 REA - I down-regulates directly CL progesterone production via P49763 REA ( 2α ) at mid - and late-luteal stages of pseudopregnancy , utilizing its cognate type I receptor with a post-receptorial mechanism that involves P98160 REA , IP₃ , DAG , PKC , P35354 REA , and NOS .

Relatively selective neuronal nitric oxide synthase inhibition by 7 - nitroindazole in monkey isolated cerebral arteries . The selectivity of 7 - nitroindazole in inhibiting endothelial and neuronal nitric oxide synthases ( P29474 REA and P29475 REA ) was investigated by comparing its inhibitory action on relaxations mediated by nitric oxide ( NO ) in response to stimulation of perivascular nerves and in response to histamine in monkey cerebral artery strips . DB02207 MEN at 2 x 10 ( - 5 ) M moderately attenuated the response to transmural electrical stimulation and to nicotine , but did to alter the endothelium-dependent relaxation in response to histamine in cimetidine-treated strips . Raising the concentration of 7 - nitroindazole to 10 ( - 4 ) M abolished the neurogenic response , partially inhibited the histamine-induced relaxation , but did not affect the response to NO . It is concluded that 7 - nitroindazole is a relatively selective P29475 REA inhibitor ; however , at high concentrations , it inhibits P29474 REA in monkey cerebral arteries .

Anticytokine treatment prevents the increase in the activity of DB00171 - ubiquitin - and Ca ( 2 + ) - dependent proteolytic systems in the muscle of tumour-bearing rats . The ascites hepatoma Yoshida AH - 130 induces loss of body weight and tissue waste . Tumour necrosis factor alpha ( P01375 REA ) plays a pivotal role in the pathogenesis of muscle wasting in this model system , but other cytokines , such as interleukin - 6 , may be involved . In order to verify whether a combined anticytokine treatment may synergistically counteract muscle protein degradation , tumour bearing rats were treated with pentoxyfilline ( PTX , an inhibitor of P01375 REA synthesis ) , or with suramin ( Q09428 REA , an antiprotozoal drug blocking the peripheral action of several cytokines including P05231 REA and P01375 REA ) , or both the drugs , and the effects on muscle proteolytic systems were assessed . Muscle protein loss in the AH - 130 - bearing rats was associated with increased activity of both the DB00171 - ubiquitin - and the calpain - dependent proteolytic pathways ( 246 % and 230 % of controls , respectively ) . Both PTX and Q09428 REA , either alone or in combination , prevented the depletion of muscle mass and significantly reduced the activity of muscle proteolytic systems . In particular , treatment with Q09428 REA , either alone or with PTX , induced a decrease in enzymatic activities to values similar to those of controls . The results obtained in the present paper demonstrate that : ( i ) muscle depletion in this model is indeed associated with increased proteasome - and calpain-dependent proteolysis , as previously suggested by increased mRNA expression of molecules pertaining to both pathways ; ( ii ) anticytokine treatments effectively reduce muscle protein loss by down-regulating the activity of at least two major proteolitic systems ; ( iii ) Q09428 REA is more effective than PTX in reducing the activity of proteolytic systems , possibly because of its multiple anticytokine action .

DB00952 MEN . The new P28222 REA / 1D agonist naratriptan , introduced in many countries in 1997 and 1998 for the acute treatment of migraine , was designed to complement the sumatriptan portfolio of offerings ( including the injection , the tablets , the nasal spray , and in some countries , the suppository ) by offering patients excellent tolerability and a sustained duration of action . Clinical studies on naratriptan , including more than 4000 patients treating more than 15,000 migraine attacks , show that naratriptan tablets 2.5 mg are distinguished from other P28222 REA / 1D agonists for migraine on the basis of their excellent tolerability profile , which does not differ from that of placebo . In addition to its tolerability , naratriptan tablets 2.5 mg possess a long duration of action with a low incidence of headache recurrence ( 17-28 % in phase II and III clinical trials ) . With its tolerability profile and long duration of action , naratriptan tablets 2.5 mg may be particularly appropriate as a single-dose alternative to NSAIDs and analgesics , which often are not effective in migraine but are used because of tolerability considerations .

Differential regulation of MeCP 2 phosphorylation in the CNS by dopamine and serotonin . Systemic administration of amphetamine ( P49418 REA ) induces phosphorylation of MeCP 2 at Ser 421 ( pMeCP 2 ) in select populations of neurons in the mesolimbocortical brain regions . Because P49418 REA simultaneously activates multiple monoamine neurotransmitter systems , here we examined the ability of dopamine ( DA ) , serotonin ( 5 - HT ) , and norepinephrine ( NE ) to induce pMeCP 2 . Selective blockade of the Q01959 REA ( Q01959 REA ) or the 5 - HT transporter ( P31645 REA ) , but not the NE transporter ( NET ) , was sufficient to induce pMeCP 2 in the CNS . Q01959 REA blockade induced pMeCP 2 in the prelimbic cortex ( P98160 REA ) and nucleus accumbens ( NAc ) , whereas P31645 REA blockade induced pMeCP 2 only in the NAc . Administration of selective DA and 5 - HT receptor agonists was also sufficient to induce pMeCP 2 ; however , the specific combination of DA and 5 - HT receptors activated determined the regional - and cell-type specificity of pMeCP 2 induction . The D ( 1 ) - class DA receptor agonist SKF 81297 induced pMeCP 2 widely ; however , coadministration of the D ( 2 ) - class agonist quinpirole restricted the induction of pMeCP 2 to GABAergic interneurons of the NAc . Intra-striatal injection of the adenylate cyclase activator forskolin was sufficient to induce pMeCP 2 in medium-spiny neurons , suggesting that the combinatorial regulation of DB02527 by different classes of DA and 5 - HT receptors may contribute to the cell-type specificity of pMeCP 2 induction . Consistent with the regulation of pMeCP 2 by multiple monoamine neurotransmitters , genetic disruption of any single monoamine transporter in Q01959 REA - , P31645 REA - , and NET-knockout mice failed to eliminate P49418 REA - induced pMeCP 2 in the NAc . Together , these studies indicate that combinatorial signaling through DA and 5 - HT receptors can regulate the brain region - and cell-type specific pMeCP 2 in the CNS .

Phase I evaluation of DB05243 MEN , an oral , potent , and selective O60674 REA inhibitor . This phase I study evaluated selective O60674 REA inhibitor DB05243 MEN in 30 patients with myelofibrosis . The initial dose cohorts were 100 , 200 , and 300 mg orally on days 1-21 of a 28 - day cycle . Central and / or peripheral neurotoxicity developed in all patients . Subsequently , patients were treated on lower doses ; neurotoxicity was again observed , leading to study termination . Peripheral neuropathy resolved in 50 % , and central neurotoxicity in all patients within months after therapy cessation . Myelosuppression was minimal . The terminal half-life of DB05243 MEN was approximately 21 h , with steady state reached by Day 8 . International Working Group defined responses were seen in three ( 10 % ) patients .

P29475 REA - dependent dilation to flow in coronary arteries of male P29474 REA - KO mice . Flow-induced dilation was examined in isolated coronary arteries of endothelial nitric oxide ( NO ) synthase knockout mice ( P29474 REA - KO ) and wild-type ( WT ) mice . The basal tone of arteries ( percentage of passive diameter ) was significantly greater in P29474 REA - KO than in WT mice ; their flow-induced dilations , however , were similar . Endothelial removal eliminated the dilations in vessels of both strains of mice . In arteries of WT mice , N ( omega ) - nitro-L-arginine methyl ester ( L-NAME ) ( 10 ( - 4 ) M ) or indomethacin ( 10 ( - 5 ) M ) alone , inhibited flow-induced dilation by approximately 50 % , whereas their simultaneous administration abolished the responses . In arteries of P29474 REA - KO mice , flow-induced dilation was inhibited by approximately 40 % with L-NAME . The residual portion ( 60 % ) of the response was eliminated by the additional administration of indomethacin . DB02207 MEN ( 10 ( - 4 ) M ) attenuated flow-induced dilation by approximately 40 % in arteries of P29474 REA - KO mice , but did not affect responses in those of WT mice . 1H - [ 1,2 , 4 ] oxadiazolo [ 4,3- a ] quinoxalin - 1 - one ( 3 x 10 ( - 5 ) M ) inhibited the L-NAME / 7 - nitroindazole-sensitive portion of the responses in arteries of P29474 REA - KO mice . Immunohistochemical evidence confirms the presence of neuronal NOS ( P29475 REA ) in the arterial endothelium of P29474 REA - KO mice . In conclusion , P29475 REA - derived NO , via activation of cGMP , together with prostaglandins , maintains flow-induced dilation in coronary arteries of male P29474 REA - KO mice .

Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 SUB or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling .

Serotonin and dopamine receptor gene polymorphisms and the risk of extrapyramidal side effects in perphenazine-treated schizophrenic patients . RATIONALE : DB00850 MENMAX DB00850 MEN , a classical antipsychotic drug , has the potential to induce extrapyramidal side effects ( EPS ) . Dopaminergic and serotonergic pathways are involved in the therapeutic and adverse effects of the drug . OBJECTIVES : To evaluate the impact of polymorphisms in the dopamine D ( 2 ) and D ( 3 ) and serotonin 2A and 2C receptor genes ( P14416 REA , P35462 REA , P28223 REA , and P28335 REA ) on short-term effects of perphenazine monotherapy in schizophrenic patients . MATERIALS AND METHODS : Forty-seven Estonian inpatients were evaluated before and after 4-6 weeks of treatment by Simpson-Angus rating scale , Barnes scale , and Positive and Negative Symptom Scale . Genotyping was performed for common P14416 REA , P35462 REA , P28223 REA , and P28335 REA gene polymorphisms , previously reported to influence receptor expression and / or function . RESULTS : Most of the patients ( n = 37 ) responded to the treatment and no significant association was observed between the polymorphisms and antipsychotic response . The 102C allele of P28223 REA and the - 697C and 23Ser alleles of P28335 REA were more frequent among patients with EPS ( n = 25 ) compared to patients without EPS ( n = 22 ) ( p = 0.02 , 0.01 , and 0.02 , respectively ) . The difference between patients with and without EPS in variant allele frequencies remained significant after multiple model analyses including age , gender , and duration of antipsychotic treatment as covariants . There was no significant association between EPS occurrence and polymorphisms in the P14416 REA and P35462 REA genes . CONCLUSIONS : An association was observed between polymorphisms in P28223 REA and P28335 REA genes and occurrence of acute EPS in schizophrenic patients treated with perphenazine monotherapy . Larger study populations are needed to confirm our findings .

DB03758 MEN - sensitive peptide binding to the N-terminal portion of P14625 REA . P14625 REA is a molecular chaperone that carries immunologically relevant peptides from cell to cell , transferring them to major histocompatibility proteins for presentation to T cells . Here we examine the binding of several peptides to recombinant P14625 REA and study the regulation and site of peptide binding . We show that P14625 REA contains a peptide-binding site in its N-terminal 355 amino acids . A number of peptides bind to this site with low on - and off-rates and with specificity that is distinct from that of another endoplasmic reticulum chaperone , P11021 REA / P11021 REA . Binding to the N-terminal fragment is sufficient to account for the peptide binding activity of the entire molecule . Peptide binding is inhibited by radicicol , a known inhibitor of the chaperone activities of HSP 90 - family proteins . However , the peptide-binding site is distinct from the radicicol-binding pocket , because both can bind to the N-terminal fragment simultaneously . Furthermore , peptide binding does not cause the same conformational change as does binding of radicicol . When the latter binds to the N-terminal domain , it induces a conformational change in the downstream , acidic domain of P14625 REA , as measured by altered gel mobility and loss of an antibody epitope . These results relate the peptide-binding activity of P14625 REA to its other function as a chaperone .

DB00227 - stimulated superinduction of P16581 REA , P05362 REA and P19320 REA in P01375 REA activated human vascular endothelial cells . Inhibitors of P04035 REA ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 REA , intercellular cell adhesion molecule - 1 ( P05362 REA ) and vascular cell adhesion molecule - 1 ( P19320 REA ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 REA . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1- 2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 REA and CAMs is correlated with a corresponding increase of selectin - and P62158 - specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque .

Dopamine modulating drugs influence striatal ( + ) - [ 11C ] DTBZ binding in rats : Q05940 REA binding is sensitive to changes in vesicular dopamine concentration . Binding of ( + ) - [ 11C ] DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 REA ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of ( + ) - [ 11C ] DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , + 14 % ) or DB01576 SUB ( d - P49418 REA , 20 mg / kg , + 12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , - 16 % ) or levodopa ( - 20 % ) . We suggest that in vivo ( + ) - [ 11C ] DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed .

[ Pharmacological and clinical profile of mitiglinide calcium hydrate ( Glufast ) , a new insulinotropic agent with rapid onset ] . DB01252 MEN calcium hydrate ( mitiglinide , Glufast ) is a new insulinotropic agent of the glinide class with rapid onset . DB01252 MEN is thought to stimulate insulin secretion by closing the DB00171 - sensitive K ( + ) ( K ( DB00171 ) ) channels in pancreatic beta-cells , and its early insulin release and short duration of action would be effective in improving postprandial hyperglycemia . In studies of various cloned K ( DB00171 ) channels , mitiglinide shows a higher selectivity for the beta-cell type of Q09428 REA / Kir 6.2 than the cardiac and smooth muscle types of K ( DB00171 ) channels in comparison with glibenclamide and glimepiride . In vitro and in vivo studies demonstrated the insulinotropic effect of mitiglinide is more potent than that of nateglinide , and mitiglinide surpassed in controlling postprandial hyperglycemia in normal and diabetic animals . In clinical trials , treatment with mitiglinide provided lasting improvement of postprandial hyperglycemia in Type 2 diabetic patients and decreased the fasting plasma glucose levels and HbA ( 1C ) values . The incidence of adverse events related to mitiglinide were nearly equivalent to placebo ; in particular there was no difference with the frequency of hypoglycemia . The results from these studies indicated that mitiglinide could be expected to possess good therapeutic features of being effective in reducing postprandial glucose excursions in the early stage of Type 2 diabetes and less incidence of events suggestive of hypoglycemia .

Exenatide twice daily : a review of its use in the management of patients with type 2 diabetes mellitus . Exenatide , administered subcutaneously twice daily ( DB01276 MEN ( ® ) ) , is a synthetic version of the natural peptide exendin - 4 , which is a glucagon-like peptide - 1 ( P0C6A0 ) receptor agonist ( incretin mimetic ) . Exenatide binds to the P43220 REA with the same affinity as P0C6A0 , but has a much longer half-life , since it is not degraded by the enzyme dipeptidyl peptidase - 4 . Exenatide twice daily enhances glucose-dependent insulin secretion , suppresses inappropriately elevated glucagon secretion , slows gastric emptying and reduces caloric intake . In well-designed clinical trials , adjunctive subcutaneous exenatide 5 or 10 μg twice daily for 16-52 weeks significantly and dose-dependently improved glycaemic control and reduced mean body weight compared with placebo in patients with type 2 diabetes inadequately controlled with oral antihyperglycaemic drugs ( OADs ) and / or basal insulin . The improvements in glycaemic control and reductions in body weight were stably maintained during long-term therapy ( up to 3.5 years ) . The efficacy of adjunctive exenatide twice daily was generally similar to that of basal , prandial or biphasic insulin , sulfonylureas , rosiglitazone and lixisenatide , and less than that of liraglutide , taspoglutide or exenatide once weekly with respect to reductions in glycated haemoglobin . Exenatide twice daily was generally well tolerated ; mild to moderate nausea and vomiting , which decreased with time on therapy , were the most common adverse events . In patients not receiving concomitant sulfonylureas or insulin , the incidence of hypoglycaemia was low ; when it did occur , it was generally mild in severity . Thus , adjunctive exenatide twice daily is a valuable option in the treatment of type 2 diabetes inadequately controlled with OADs and / or basal insulin .

Serotonergic regulation of somatosensory cortical development : lessons from genetic mouse models . Monoaminergic neurotransmitter systems appear early during embryogenesis , suggesting that they could play important roles in brain development . Accumulated evidence indicates that serotonin ( 5 - hydroxytryptamine , 5 - HT ) regulates neural as well as nonneural development , including early aspects of embryonic development , differentiation of neuronal progenitors , and morphogenesis of the craniofacial region , heart and limb . Recent studies using monoamine oxidase-A ( P21397 REA ) , 5 - HT transporter , vesicular monoamine transporter - 2 ( Q05940 REA ) and P28222 REA receptor single , double and triple knockout mice have provided evidence that the serotonergic system plays important roles in barrel field formation in the developing somatosensory cortex . Here we review evidence from these genetic mouse models and , based on the accumulated evidence , propose a testable model for future studies of mechanisms underlying serotonergic regulation of cortical development .

Genetic basis of delay discounting in frequent gamblers : examination of a priori candidates and exploration of a panel of dopamine-related loci . INTRODUCTION : Delay discounting is a behavioral economic index of impulsivity that reflects preferences for small immediate rewards relative to larger delayed rewards . It has been consistently linked to pathological gambling and other forms of addictive behavior , and has been proposed to be a behavioral characteristic that may link genetic variation and risk of developing addictive disorders ( i . e . , an endophenotype ) . Studies to date have revealed significant associations with polymorphisms associated with dopamine neurotransmission . The current study examined associations between delay discounting and both previously linked variants and a novel panel of dopamine-related variants in a sample of frequent gamblers . METHODS : Participants were 175 weekly gamblers of European ancestry who completed the Monetary Choice Questionnaire to assess delay discounting preferences and provided a DNA via saliva . RESULTS : In a priori tests , two loci previously associated with delayed reward discounting ( rs1800497 and rs4680 ) were not replicated , however , the long form of P21917 REA VNTR was significantly associated with lower discounting of delayed rewards . Exploratory analysis of the dopamine-related panel revealed 11 additional significant associations in genes associated with dopamine synthesis , breakdown , reuptake , and receptor function ( P35462 REA , Q01959 REA , DDC , P09172 REA , and Q05940 REA ) . An aggregate genetic risk score from the nominally significant loci accounted for 17 % of the variance in discounting . Mediational analyses largely supported the presence of indirect effects between the associated loci , delay discounting , and pathological gambling severity . CONCLUSIONS : These findings do not replicate previously reported associations but identify several novel candidates and provide preliminary support for a systems biology approach to understand the genetic basis of delay discounting .

P01375 REA inhibits flow and insulin signaling leading to NO production in aortic endothelial cells . Endothelial cells release nitric oxide ( NO ) acutely in response to increased " flow " or fluid shear stress ( FSS ) , and the increase in NO production is correlated with enhanced phosphorylation and activation of endothelial nitric oxide synthase ( P29474 REA ) . Both vascular endothelial growth factor and FSS activate endothelial protein kinase B ( P31749 REA ) by way of incompletely understood pathway ( s ) , and , in turn , P31749 REA phosphorylates P29474 REA at DB00133 - 1179 , causing its activation . In this study , we found that either FSS or insulin stimulated insulin receptor substrate - 1 ( P35568 REA ) tyrosine and serine phosphorylation and increased P35568 REA - associated phosphatidylinositol 3 - kinase activity , phosphorylation of P31749 REA DB00133 - 473 , phosphorylation of P29474 REA DB00133 - 1179 , and NO production . Brief pretreatment of bovine aortic endothelial cells with tumor necrosis factor-alpha ( P01375 REA ) inhibited the above described FSS - or insulin-stimulated protein phosphorylation events and almost totally inhibited FSS - or insulin-stimulated NO production . These data indicate that FSS and insulin regulate P29474 REA phosphorylation and NO production by overlapping mechanisms . This study suggests one potential mechanism for the development of endothelial dysfunction in disease states with alterations in insulin regulation and increased P01375 REA levels .

DB04900 improves severe acute pancreatitis in rats via regulation of peripheral T cell number and cytokine serum level . AIM : The aim of this study was to investigate the effect of thymosin alpha 1 ( Q96RJ0 ) on severe acute pancreatitis ( SAP ) in rats . METHODS : Healthy Sprague-Dawley rats ( n = 72 ) were randomly divided into four groups : control group , SAP group , and two Q96RJ0 treated groups . SAP was induced by injection of 5 % sterile sodium taurocholate into the biliopancreatic duct ( BPD ) , after which Q96RJ0 was given subcutaneously at 0 and 2 h at a dose of 100 microg / kg . The rats were killed at 3 , 6 and 12 h , respectively . Serum amylase and lipase , interleukin ( IL ) - 1beta , tumor necrosis factor-alpha ( P01375 REA ) , pancreatic wet / dry weight ratio and the percentage of CD3 / P01730 REA + / CD8 + T cells in peripheral blood mononuclear cells ( PBMC ) were measured . Next , 30 rats were randomly divided into three groups ( each group containing 10 animals ) : SAP group ( S ) and two Q96RJ0 treated groups . The effects of Q96RJ0 on the survival of SAP were assessed 72 h after the induction of SAP . RESULTS : There was no significant change in the serum amylase and lipase levels after Q96RJ0 administration . Levels of serum IL - 1beta , P01375 REA and pancreatic wet / dry weight ratio were significantly reduced after Q96RJ0 - treatment . Application of Q96RJ0 significantly balanced CD3 / P01730 REA + / CD8 + T cells of PBMC and improved histological scores and the survival rate . CONCLUSION : Q96RJ0 can reduce pancreatic inflammation by regulating differentiation of CD3 / P01730 REA + T cells and decreasing the release of cytokines , thus attenuates pancreatic severity in SAP rats .

DB00952 MEN mitigates CGRP 1 - associated motor neuron degeneration caused by an expanded polyglutamine repeat tract . Spinal and bulbar muscular atrophy ( SBMA ) is a motor neuron disease caused by the expansion of the CAG triplet repeat within the androgen receptor ( AR ) gene . Here , we demonstrated that pathogenic AR upregulates the gene encoding calcitonin gene-related peptide α ( CGRP 1 ) . In neuronal cells , overexpression of CGRP 1 induced cellular damage via the activation of the c-Jun N-terminal kinase ( JNK ) pathway , whereas pharmacological suppression of CGRP 1 or JNK attenuated the neurotoxic effects of pathogenic AR . The depletion of CGRP 1 inactivated JNK and suppressed neurodegeneration in a mouse model of SBMA . DB00952 MEN , a serotonin 1B / 1D ( 5 - hydroxytryptamine 1B / 1D , or P28222 REA / 1D ) receptor agonist , decreased CGRP 1 expression via the induction of dual-specificity protein phosphatase 1 ( P28562 REA ) , attenuated JNK activity and mitigated pathogenic AR-mediated neuronal damage in cellular and mouse SBMA models . These observations suggest that pharmacological activation of the P28222 REA / 1D receptor may be used therapeutically to treat SBMA and other polyglutamine-related neurodegenerative diseases .

Novel P0C6A0 mimetics developed to treat type 2 diabetes promote progenitor cell proliferation in the brain . One of the symptoms of diabetes is the progressive development of neuropathies . One mechanism to replace neurons in the CNS is through the activation of stem cells and neuronal progenitor cells . We have tested the effects of the novel P0C6A0 mimetics exenatide ( exendin - 4 ; DB01276 MEN ) and liraglutide ( DB06655 ; DB06655 ) , which are already on the market as treatments for type 2 diabetes , on the proliferation rate of progenitor cells and differentiation into neurons in the dentate gyrus of brains of mouse models of diabetes . P0C6A0 analogues were injected subcutaneously for 4 , 6 , or 10 weeks once daily in three mouse models of diabetes : ob / ob mice , db / db mice , or high-fat-diet-fed mice . Twenty-four hours before perfusion , animals were injected with 5 ' - bromo - 2 ' - deoxyuridine ( BrdU ) to mark dividing progenitor cells . By using immunohistochemistry and stereological methods , the number of progenitor cells or doublecortin-positive young neurons in the dentate gyrus was estimated . We found that , in all three mouse models , progenitor cell division was enhanced compared with nondiabetic controls after chronic i . p . injection of either liraglutide or exendin - 4 by 100-150 % ( P < 0.001 ) . We also found an increase in young neurons in the DG of high-fat-diet-fed mice after drug treatment ( P < 0.001 ) . The P43220 REA antagonist exendin ( 9-36 ) reduced progenitor cell proliferation in these mice . The results demonstrate that P0C6A0 mimetics show promise as a treatment for neurodegenerative diseases such as Alzheimer ' s disease , because these novel drugs cross the blood-brain barrier and increase neuroneogenesis .

P36888 REA - mutant allelic burden and clinical status are predictive of response to P36888 REA inhibitors in AML . We examined 6 different P07333 REA - like tyrosine kinase - 3 ( P36888 REA ) inhibitors ( lestaurtinib , midostaurin , DB05213 MEN , KW - 2449 , sorafenib , and sunitinib ) for potency against mutant and wild-type P36888 REA , as well as for cytotoxic effect against a series of primary blast samples obtained from patients with acute myeloid leukemia ( AML ) harboring internal tandem duplication ( P36888 REA / ITD ) mutations . We found that inhibition of P36888 REA autophosphorylation in a P36888 REA / ITD specimen does not always induce cell death , suggesting that some P36888 REA / ITD AML may not be addicted to P36888 REA signaling . Relapsed samples and samples with a high mutant allelic burden were more likely to be responsive to cytotoxicity from P36888 REA inhibition compared with the samples obtained at diagnosis or those with a low mutant allelic burden . These P36888 REA inhibitors varied to a considerable degree in their selectivity for P36888 REA , and this selectivity influenced the cytotoxic effect . These results have important implications for the potential therapeutic use of P36888 REA inhibitors in that patients with newly diagnosed P36888 REA - mutant AML might be less likely to respond clinically to highly selective P36888 REA inhibition .

DB00188 prevents the expression of P45452 REA and the degradation of collagen type 2 in human chondrocytes . The structural backbone of extracellular matrix in cartilage is the collagen fibril , which is mainly composed of type II collagen . A measurable increase in type II collagen denaturation and degradation has been found in early Osteoarthritis ( OA ) . Pro-inflammatory cytokine such as P01375 REA - α produced in OA cartilage induced the expression of matrix metalloproteinase - 13 ( P45452 REA ) , which targets and degrades type II collagen . DB00188 is a proteasome inhibitor approved by the FDA for treatment of multiple myeloma and mantel cell lymphoma . The effects of bortezomib in OA have not been reported before . In this study , we found that bortezomib is able to suppress the degradation of type II collagen induced by P01375 REA - α in human chondrocytes . Mechanistically , bortezomib treatment inhibits the expression of P10914 REA through blunting O60674 REA / P42224 REA pathway , thereby prevents the induction of P45452 REA as well as the degradation of type II collagen . Our findings suggest the therapeutic potentials of bortezomib in patients with OA .

P36888 REA inhibition as therapy in acute myeloid leukemia : a record of trials and tribulations . Acute myeloid leukemia ( AML ) is a hematologic malignancy with a poor prognosis . Approximately one quarter of the patients with AML also carry an internal tandem duplication ( ITD ) mutation in the gene encoding P07333 REA - like tyrosine kinase 3 ( P36888 REA ) , which has a significantly deleterious impact on prognosis . The ITD mutation renders P36888 REA constitutively active and leads to uncontrolled proliferation of the leukemic blast . Over the course of the last decade , a variety of compounds have been developed in preclinical and clinical studies as potent inhibitors of P36888 REA . Many of the earlier agents under investigation , such as lestaurtinib , midostaurin , and sunitinib , were initially developed as inhibitors of other tyrosine kinases and as targeted therapies in a variety of malignancies . These compounds have been demonstrated to have some efficacy in clinical trials of AML , mainly manifesting as transient decreases in circulating blasts correlating with effective in vivo suppression of the P36888 REA target . Nevertheless , the cumbersome pharmacokinetics of some compounds and the suboptimal specificity and potency of others have limited their therapeutic efficacy . In the last few years , newer , more potent and specific agents have been under investigation , with the leading example being DB05213 MEN . This agent has shown significant promise in early phases of clinical investigation , and is currently in more advanced clinical trials . Hope remains that P36888 REA inhibition will be become an effective therapeutic adjunct to our current treatment approach to AML .

Effects of mitiglinide ( S 21403 ) on Kir 6.2 / Q09428 REA , Kir 6.2 / SUR 2A and Kir 6.2 / SUR 2B types of DB00171 - sensitive potassium channel . 1 . We have investigated the mechanism of action of the novel anti-diabetic agent mitiglinide ( S 21403 ) on Kir 6.2 / Q09428 REA , Kir 6.2 / SUR 2A and Kir 6.2 / SUR 2B types of DB00171 - sensitive potassium ( K ( DB00171 ) ) channel . These possess a common pore-forming subunit , Kir 6.2 , and different regulatory sulphonylurea receptor ( Q09428 REA ) subunits . It is believed that they correspond to native K ( DB00171 ) channels in pancreatic beta-cells , heart and non-vascular smooth muscle , respectively . 2 . Kir 6.2 was coexpressed with Q09428 REA , SUR 2A or SUR 2B in Xenopus oocytes and macroscopic currents were recorded in giant inside-out membrane patches . DB01252 MEN was added to the intracellular membrane surface . 3 . DB01252 MEN inhibited Kir 6.2 / Q09428 REA currents at two sites : a low-affinity site on Kir 6.2 and a high-affinity site on Q09428 REA . Low-affinity inhibition was similar for all three types of K ( DB00171 ) channel but high-affinity inhibition was greater for Kir 6.2 / Q09428 REA currents ( IC ( 50 ) , 4 nM ) than for Kir 6.2 / SUR 2A or Kir 6.2 / SUR 2B currents ( IC ( 50 ) , 3 and 5 microM , respectively ) . 4 . Inhibition of Kir 6.2 / Q09428 REA currents was only slowly reversible on the time scale of electrophysiological experiments . 5 . Kir 6.2 / Q09428 REA - S1237Y currents , which previously have been shown to lack high affinity tolbutamide inhibition , resembled Kir 6.2 / SUR 2 currents in being unaffected by 100 nM but blocked by 10 microM mitiglinide . 6 . Our results show that mitiglinide is a high-affinity drug that shows a 1000 fold greater affinity for the beta-cell type than the cardiac and smooth muscle types of K ( DB00171 ) channel , when measured in excised patches .

Transduction of P28906 REA + cells with lentiviral vectors enables the production of large quantities of transgene-expressing immature and mature dendritic cells . BACKGROUND : Genetically engineered dendritic cells ( DC ) presenting specific antigens to T cells may be of great interest for immunotherapy . For this reason , the production of transgene-expressing DC derived from P28906 REA + cells transduced either shortly after ex vivo purification or during their differentiation into DC were evaluated . METHODS : P28906 REA + cells were transduced with lentivectors encoding for GFP before or after 21 days of culture with P36888 REA - ligand , thrombopoietin and stem cell factor and induction into DC with GM - P04141 REA + P05112 REA ( G4 ) or G4 + P01375 REA ( GT4 ) . GFP and DC-specific marker expression was assessed by flow cytometry , and allostimulatory capacity was evaluated on GFP + and GFP - sorted cells . RESULTS : Immature ( G4 - induced ) DC obtained from amplified P28906 REA + cells were transducible by lentiviral vectors while mature ( GT4 - induced ) DC were rather refractory . Moreover , since differentiated DC did not proliferate , large quantities of vectors were required to generate transgene-expressing cells with this protocol . In contrast , greater numbers of both immature and mature GFP - expressing DC were obtained with P28906 REA + cells exposed to lentivector shortly after purification . By the time of DC induction , GFP + cells had increased by approximately 170 - fold . After DC induction with G4 , 32 % of CD1a + , HLA-DR + , or P25942 REA + cells expressed GFP . CD1a + P12830 REA + GFP + Langerhans-like DC were also obtained . Incubation with P01375 REA induced mature Q01151 + GFP + DC that displayed a higher allostimulatory capacity than cells induced with G4 alone . CONCLUSION : The transduction of a small number of P28906 REA + cells with minimal doses of lentivector may allow for the production of a large number of DC expressing selected antigens useful for immunotherapy .

Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 REA mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 REA expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR 12909 , serotonin , mazindol , nomifensin and DB01576 SUB , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 REA in packaging monoamine transmitters including histamine .

Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 - R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 REA ) in the presence of cycloheximide ( Reid , T . R . , Torti , F . , and Ringold , G . M . ( 1989 ) J . Biol . Chem . 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 REA or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 REA - resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6 - desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 - R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 REA in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 REA and is essential to the rapid cytotoxic response elicited by P01375 REA in the absence of protein synthesis in wild-type Q96RJ0 cells .