MH_dev_148

P35372 REA - dependent and independent components in effects of tramadol . DB00193 SUB is thought to induce analgesia via both opioid and non-opioid pathways , although the precise mechanisms remain to be elucidated . In this study , we investigated the roles of the mu-opioid receptor ( MOP ) in analgesic and rewarding effects of tramadol by using MOP knockout ( KO ) mice . DB00193 SUB - induced antinociception , assessed by hot-plate and tail-flick tests , was significantly reduced in heterozygous and homozygous MOP-KO mice when compared with that in wild-type mice . Interestingly , however , tramadol retained its ability to induce significant antinociception in homozygous MOP-KO mice . The tramadol-induced antinociception remaining in homozygous MOP-KO mice was not significantly affected by methysergide , a serotonin receptor antagonist , but was partially blocked by yohimbine , an adrenaline alpha 2 receptor antagonist , and both naloxone , a non-selective opioid receptor antagonist , and yohimbine . In addition , antinociceptive effects of an active tramadol metabolite M1 were abolished or remarkably reduced in MOP-KO mice . On the other hand , neither wild-type nor homozygous MOP-KO mice showed significant place preference for tramadol in a conditioned place preference test , although there were slight tendencies toward preference in wild-type mice and avoidance in homozygous MOP-KO mice . These results strongly support the idea suggested in the previous pharmacological studies that MOP and the adrenaline alpha 2 receptor mediate most of the analgesic properties of tramadol .

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 MEN , 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 .

Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 REA ( P05231 REA ) , C-Reactive Protein ( CRP ) , P00734 REA Fragments 1 and 2 ( F 1 + 2 ) , cortisol and P00747 REA Activator Inhibitor 1 ( P05121 REA ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 REA and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 REA level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge .

Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 REA , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 REA ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 REA ) 293 cells co-expressing P35372 REA and O14939 REA , treatment with ( D-Ala 2 , Me Phe 4 , Glyol 5 ) enkephalin ( DAMGO ) led to an increase in O14939 REA activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 REA . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR 1D , also termed MOP ( 1D ) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR 1D also mediates an agonist-independent ( constitutive ) O14939 REA - activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 REA activity by over-expression of a dominant negative O14939 REA ( nPLD 2 ) blocked the constitutive O14939 REA activation and impaired the endocytosis of MOR 1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 REA ) is also mediated by a O14939 REA - dependent pathway . These data indicate the generally important role for O14939 REA in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis .

[ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 - sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2- 20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2 + . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W - 5 and W - 7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2 + and blocked the effect at higher concentrations of Ca2 + . DB00477 MENMAX DB00477 MEN , another calmodulin antagonist , reduced Ca2 + - stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 REA and DB02527 increased in the presence of 5 microM Ca2 + . AC stimulating effects of P01133 REA , DB02527 and insulin decreased in the presence of 100 microM Ca2 + , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2 + and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T . pyriformis which mediate enzyme stimulation by P01133 REA , DB02527 , insulin , and serotonin .

Clinical pharmacology of serotonin-altering medications for decreasing alcohol consumption . Variations in serotonin neurotransmission influence alcohol consumption ( AC ) . Levels of 5 - HT and metabolites are low in some brain regions of alcohol preferring rats and in P04141 REA of alcoholics . Pharmacological treatments which enhance serotonergic neurotransmission ( uptake inhibitors , releasers , agonists ) consistently reduce AC in rats . Serotonin uptake inhibitors ( SUI ; e . g . , citalopram , fluoxetine ) have been studied extensively in humans . In several double-blind randomized , placebo-controlled clinical trials , SUI have consistently decreased AC by averages of 15 % to 20 % in nondepressed mildly / moderately dependent alcoholics who received no other treatment . Effects were dose-dependent and not related to side effects ( few and mild ) or changes in anxiety or depression ( not observed ) . SUI also decreased desire to drink and liking for alcohol , thus suggesting a mechanism for effects . Other drugs acting on the 5 - HT system have been tested in humans , but results are difficult to interpret . For example , buspirone , a P08908 REA receptor partial agonist , reduced anxiety and alcohol craving , but not AC ; a 5 - HT partial agonist , m-CPP , increased alcohol craving in abstinent alcoholics ; modest reductions in AC were observed with a 5 - Q9H205 REA antagonist , ondansetron ( 0.5 mg / day , but not 4 mg / day ) . The therapeutic potentials of these medications are being studied . For example , SUI effects on AC were enhanced by a brief psychosocial intervention . Since SUI decrease urge to drink , they may be suitable pharmacological adjuncts in relapse prevention strategies . SUI and other serotonin-altering medications are promising new neuropharmacological treatments for reducing AC .

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 MEN 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 .

Single-prolonged stress induce changes of P62158 / CaMKIIα in the rats of dorsal raphe nucleus . Ca2 + / calmodulin-dependent protein kinase IIα ( CaMKIIα ) is identified as a Ca2 + - dependent kinase in brain involved in the activation of DB00150 hydroxylase ( P17752 REA ) acting through direct phosphorylation of P17752 REA , and playing key roles in the signaling pathways initiated by various G protein-coupled 5 - HT receptors . The goal of this study is to detect whether there are changes of P62158 and CaMKIIα in dorsal raphe nucleus in the rats exposed to single-prolonged stress ( P49903 ) , which is a model employed in post-traumatic stress disorder ( PTSD ) study extensively . A total of 90 male Wistar rats were randomly divided into a normal control group and P49903 groups of 7d , 14d . The changes of P62158 / CaMKIIα were detected by immunohistochemistry , reverse transcription-polymerase chain reaction and western blot . Our results demonstrate that both expressions of P62158 and CaMKIIα significantly increase ( P < 0.001 ) in the P49903 7d group than that in the control group , and then decreased dramatically ( P < 0.001 ) 14 days after P49903 . Our results confirm that P49903 induce changes of P62158 / CaMKIIα in the dorsal raphe nucleus . Changes of P62158 / CaMKIIα may be associated with the activation of P08908 REA receptor , and may contribute to the progress of molecular mechanism of PTSD .

Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 REA ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 REA in the actions of a 5 - HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression / antidepressant response ; and second , by examining the role of the P08908 REA receptor subtype in the regulation of P15692 REA , and the cellular localization of antidepressant regulation of P15692 REA expression . The results show that pharmacological inhibition of P15692 REA receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 REA - Flk - 1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 REA receptors is sufficient to induce P15692 REA expression and that a P08908 REA antagonist blocks both the increase in P15692 REA and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 REA expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 REA is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 REA receptors located on neurons and endothelial cells .

P00734 REA in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 REA ) has not been investigated . We determined the concentration of prothrombin in P04141 REA with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit - - 0.7 ng / ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 REA was 0.55 mg / l ( CV + / - 33 % , range : 0.28- 0.93 mg / l ) , in normal plasma 121.8 mg / l + / - 21 % , resulting in a mean P04141 REA / plasma concentration quotient ( Q ( Proth ) - - 4.5 x 10 ( - 3 ) ( CV + / - 35 % , range : 2.1- 8.3 x 10 ( - 3 ) ) corresponding to a mean albumin quotient in this group of subjects of Q ( Alb ) = 5.8 x 10 ( - 3 ) . Due to the Q ( Proth ) and the molecular weight of prothrombin ( 72 kDa ) - - similar to that of albumin - - we conclude that prothrombin in normal human P04141 REA originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 REA is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 REA if the blood - P04141 REA barrier function is normal .

Candidate genes for cannabis use disorders : findings , challenges and directions . AIM : Twin studies have shown that cannabis use disorders ( abuse / dependence ) are highly heritable . This review aims to : ( i ) review existing linkage studies of cannabis use disorders and ( ii ) review gene association studies , to identify potential candidate genes , including those that have been tested for composite substance use disorders and ( iii ) to highlight challenges in the genomic study of cannabis use disorders . METHODS : Peer-reviewed linkage and candidate gene association studies are reviewed . RESULTS : Four linkage studies are reviewed : results from these have homed in on regions on chromosomes 1 , 3 , 4 , 9 , 14 , 17 and 18 , which harbor candidates of predicted biological relevance , such as monoglyceride lipase ( Q99685 REA ) on chromosome 3 , but also novel genes , including Q9HBW9 [ epidermal growth factor ( P01133 REA ) , latrophilin and seven transmembrane domain containing 1 ] on chromosome 1 . Gene association studies are presented for ( a ) genes posited to have specific influences on cannabis use disorders : P21554 REA , CB2 , FAAH , Q99685 REA , Q8NER1 and Q9Y2T6 REA and ( b ) genes from various neurotransmitter systems that are likely to exert a non-specific influence on risk of cannabis use disorders , e . g . P47869 REA , P14416 REA and P35372 REA . CONCLUSIONS : There are challenges associated with ( i ) understanding biological complexity underlying cannabis use disorders ( including the need to study gene-gene and gene-environment interactions ) , ( ii ) using diagnostic versus quantitative phenotypes , ( iii ) delineating which stage of cannabis involvement ( e . g . use versus misuse ) genes influence and ( iv ) problems of sample ascertainment .

P35372 REA expression is increased in inflammatory bowel diseases : implications for homeostatic intestinal inflammation . BACKGROUND AND AIMS : Recent studies with mu opioid receptor ( MOR ) deficient mice support a physiological anti-inflammatory effect of MOR at the colon interface . To better understand the potential pharmacological effect of certain opiates in inflammatory bowel diseases ( Q9UKU7 ) , we ( 1 ) evaluated the regulation in vivo and in vitro of human MOR expression by inflammation ; and ( 2 ) tested the potential anti-inflammatory function of a specific opiate ( DALDA ) in inflamed and resting human mucosa . PATIENTS AND METHODS : Expression of MOR mRNA and protein was evaluated in healthy and inflamed small bowel and colonic tissues , isolated peripheral blood mononuclear cells and purified monocytes , and P01730 REA + and CD8 + T cells from healthy donors and Q9UKU7 patients . The effect of cytokines and nuclear factor kappaB ( NFkappaB ) activation on MOR expression in lymphocyte T and monocytic human cell lines was assessed . Finally , DALDA induced anti-inflammatory effect was investigated in mucosal explants from controls and Q9UKU7 patients . RESULTS : MOR was expressed in ileal and colonic enteric neurones as well as in immunocytes such as myeloid cells and P01730 REA + and CD8 + T cells . Overexpressed in active Q9UKU7 mucosa , MOR was significantly enhanced by cytokines and repressed by NFkappaB inhibitor in myeloid and lymphocytic cell lines . Furthermore , ex vivo DALDA treatment dampened tumour necrosis factor alpha mRNA expression in the colon of active Q9UKU7 patients . CONCLUSIONS : Given the increased expression of MOR and the ex vivo beneficial effect of DALDA in active Q9UKU7 , natural and / or synthetic opioid agonists could help to prevent overt pathological intestinal inflammation .

Cloning , after cloning , knock-out mice , and physiological functions of MAO A and B . Cloning of MAO A and B has demonstrated clearly that MAO A and B are coded by different proteins with 70 % amino acid identity . With the MAO A and B cDNA clones , we showed the tissue distribution and genomic structure of MAO A and B , the latter suggesting that they are derived from the same ancestral gene . The active sites , the role of cysteine residues , the three-dimensional models and the mitochondria targeting domains of both isoenzymes have been established . The transcriptional regulation of MAO A and B has been studied . MAO A KO mice showed increased levels of serotonin ( 5 - HT ) , norepinephrine ( NE ) , dopamine ( DA ) whereas MAO B KO mice showed increased phenylethylamine ( PEA ) levels only . Both MAO A and B KO mice showed increased response to stress . MAO A KO mice showed increased emotional learning and memory and aggressive behavior , but the vesicular monoamine transporter ( Q05940 REA ) , P08908 REA , 5 - Q13049 REA and P28335 REA receptors were down regulated . 5 - Q13049 REA antagonist , ketanserin and MDL 100907 were able to abolish the aggression , suggesting that the aggressive behavior may be mediated by 5 - Q13049 REA receptor . In contrast , MAO B KO mice are resistant to MPTP , a toxin which induces Parkinson ' s syndromes . Studies of these mice suggest that MAO A and B have distinct biochemical and physiological functions .

Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB / Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB / Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 REA ( IKK ) activity was investigated using purified recombinant O15111 REA and - beta proteins . RESULTS : NF-kappaB / Rel activity induced by tumor necrosis factor alpha , 12 - O-tetradecanoylphorbol - 13 - acetate , or overexpression of NF-kappaB-inducing kinase , O15111 REA , O14920 REA , or constitutively active O15111 REA and O14920 REA mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 REA and O14920 REA in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 MEN had no effect . Activation of extracellular signal-related kinase ( P29323 REA ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 REA and - beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 REA and - beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine .

Arsenic decreases antinociceptive activity of paracetamol : possible involvement of serotonergic and endocannabinoid receptors . We assessed whether repeated arsenic exposure can decrease paracetamol-mediated antinociception by modulating serotonergic and endocannabinoid pathways . Rats were preexposed to elemental arsenic ( 4ppm ) as sodium arsenite through drinking water for 28 days . Next day paracetamol ' s ( 400mg / kg , oral ) antinociceptive activity was assessed through formalin-induced nociception . Serotonin content and gene expression of P08908 REA , 5 - Q13049 REA and P21554 REA receptors were evaluated in brainstem and frontal cortex . Arsenic decreased paracetamol-mediated analgesia . DB00316 , but not arsenic , increased serotonin content in these regions . Arsenic attenuated paracetamol-mediated increase in serotonin level . DB00316 did not alter P08908 REA expression , but caused down-regulation of 5 - Q13049 REA and up-regulation of P21554 REA receptors . Arsenic down-regulated these receptors . However , paracetamol-mediated down-regulation of 5 - Q13049 REA was more pronounced . Arsenic did not modify paracetamol ' s effect on P08908 REA expression , but reduced paracetamol-mediated down-regulation of 5 - Q13049 REA and reversed up-regulation of P21554 REA receptors . Results suggest arsenic reduced paracetamol-induced analgesia possibly by interfering with pronociceptive 5 - Q13049 REA and antinociceptive P21554 REA receptors .

Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 REA receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 REA , CD8 and forkhead box P09131 ( Foxp 3 ) , respectively . P14136 REA , Iba 1 and P34810 REA - immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 REA receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 REA receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed .

DB01095 MEN inhibits growth and alters the malignant phenotype of the P13671 REA glioma cell line . BACKGROUND : DB01095 MEN is a member of the family of P04035 REA inhibitors ( statins ) extensively used in medical practice . Increasing evidence suggests that fluvastatin may be implicated in suppression of cancer growth and development . The aim of the present study was to investigate the anti-cancer potential of fluvastatin in P13671 REA rat malignant glioma cells . METHODS : First , the effects of fluvastatin on cell viability ( MTT assay ) , proliferation ( BrdU assay ) , cell morphology , and cytoskeleton were examined . Subsequently , its effect on extracellular signal regulated kinase 1 and 2 ( P27361 REA / 2 ) and P45983 REA and 2 ( JNK 1/2 ) expression was estimated by Western blot . Finally , the influence of fluvastatin on cell migration and production of P14780 REA and P15692 REA was determined using a wound-healing assay and ELISA test , respectively . RESULTS : The results obtained showed that fluvastatin had a remarkable inhibitory and cytotoxic effect on tumor P13671 REA cells ( IC ( 50 ) = 8.6 μM , 48 h ) , but did not inhibit the growth of normal neuronal cells . The concentrations from 1 to 10 μM induced marked morphologic alterations typical for apoptosis including shrinkage of cytoplasm , chromatin condensation , and nucleus breakdown . CONCLUSION : The inhibitory effects of fluvastatin on cell proliferation seemed to be associated with decreased p - P27361 REA / 2 expression , upregulation of p - P45983 REA / 2 , and reduction in the P14780 REA and P15692 REA concentrations in culture media . The high anticancer ( antiproliferative , proapoptotic , antiinvasive ) activity of fluvastatin and lack of its toxicity against normal cells indicate a potential use of this statin in the treatment of malignant glioma .

DB00193 SUB and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 REA ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 REA ) - deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5 - hydroxy-l-tryptophan ( 5 - HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 REA wild-type ( + / + ) , heterozygous ( + / - ) and knockout ( - / - ) mice . Comparisons were made with P31645 REA mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 REA . The exaggerated response to tramadol in P31645 REA - / - mice was blocked by pretreatment with the P08908 REA antagonist WAY 100635 . Further , we show that morphine - , meperidine - and tramadol-induced analgesia is markedly decreased in P31645 REA - / - mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 REA polymorphisms that may reduce P31645 REA by more than 50 % , the level in P31645 REA + / - mice .

Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase - 5 ( O76074 REA ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 REA inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 REA ) or bilateral P21554 REA , and were left untreated or given retrolingually 5 mg / kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 MEN treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 REA compared with sham-operated rats . DB00820 MEN also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC / collagen , and replication index , and improved the lower collagen III / I ratio and the increase in apoptotic index , caused by P21554 REA , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta 1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction .

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 .

Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5 - fluoruracil-based chemotherapy regimens was analyzed for variants of IL - 1B , P60568 REA , P05231 REA , IL - 6R , P22301 REA , P01375 REA , TGF-b , O60603 REA , O00206 REA , Q9Y6Y9 REA , Q99836 REA , P23560 REA , CRP , ICE , and P35372 REA . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 REA and P01375 REA genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population .

DB00278 MEN - coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 MEN has a high affinity for thrombin and its thrombin binding is reversible . P00734 REA derived from a Ba ( 2 + ) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days .

Genetic contributions to labor pain and progress . Studies on genetic contributions to labor analgesia have essentially evaluated the μ-opioid receptor gene ( P35372 REA ) , with some evidence that p . 118A / G of P35372 REA influences the response to neuraxial opioids . As for labor progress , the β2 - adrenergic receptor gene ( P07550 REA ) is associated with preterm labor and delivery , and impacts the course of labor . Taken together though , there is no evidence that pharmacogenetic testing is needed or beneficial in the context of obstetric anesthesia ; however , realizing the influence of genetic variants on specific phenotypes provides the rationale for a more cautious interpretation of clinical studies that attempt to find a dose-regimen that fits all .

P35372 REA activation of P27361 REA / 2 is P35626 REA and arrestin dependent in striatal neurons . In this study we investigated the mechanisms responsible for Q96HU1 kinase P27361 REA / 2 activation following agonist activation of endogenous mu opioid receptors ( MOR ) normally expressed in cultured striatal neurons . Treatment with the MOR agonist fentanyl caused significant activation of P27361 REA / 2 in neurons derived from wild type mice . Fentanyl effects were blocked by the opioid antagonist naloxone and were not evident in neurons derived from MOR knock-out ( - / - ) mice . In contrast , P27361 REA / 2 activation by fentanyl was not evident in neurons from P35626 REA - / - mice or neurons pretreated with small inhibitory RNA for arrestin 3 . Consistent with this observation , treatment with the opiate morphine ( which is less able to activate arrestin ) did not elicit P27361 REA / 2 activation in wild type neurons ; however , transfection of arrestin 3 - ( R170E ) ( a dominant positive form of arrestin that does not require receptor phosphorylation for activation ) enabled morphine activation of P27361 REA / 2 . In addition , activation of P27361 REA / 2 by fentanyl and morphine was rescued in P35626 REA - / - neurons following transfection with dominant positive arrestin 3 - ( R170E ) . The activation of P27361 REA / 2 appeared to be selective as p38 Q96HU1 kinase activation was not increased by either fentanyl or morphine treatment in neurons from wild type , MOR - / - , or P35626 REA - / - mice . In addition , U0126 ( a selective inhibitor of MEK kinase responsible for P29323 REA phosphorylation ) blocked P27361 REA / 2 activation by fentanyl . These results support the hypothesis that MOR activation of P27361 REA / 2 requires opioid receptor phosphorylation by P35626 REA and association of arrestin 3 to initiate the cascade resulting in P27361 REA / 2 phosphorylation in striatal neurons .

DB05039 MEN inhibits tumor cell invasiveness and P14780 REA expression by suppressing IKK / NF-κB activation . The β2 adrenergic receptor ( P07550 REA ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 REA ) . Although a number of P07550 REA agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2 - agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 REA patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin - 2 mediates the internalization of P07550 REA following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 REA - α ) - induced NF-κB activity by reducing levels of both phosphorylated-IKK and - IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 REA , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 REA - α / NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin 2 - dependent manner , preventing further lung damage and improving lung function in P48444 REA patients .

mu-Opioid receptor agonists differentially regulate the expression of miR - 190 and Q13562 REA . The agonists of mu-opioid receptor ( P35372 REA ) induce extracellular signal-regulated kinase ( P29323 REA ) phosphorylation through different pathways : morphine uses the protein kinase C ( PKC ) - pathway , whereas fentanyl functions in a beta-arrestin 2 - dependent manner . In addition , the two pathways result in the different cellular location of phosphorylated P29323 REA and the activation of different sets of transcriptional factors . In the current study , the influence of the two pathways on the expression of microRNAs ( miRNAs ) was investigated . After treating the primary culture of rat hippocampal neurons and the mouse hippocampi with morphine or fentanyl for 3 days , seven miRNAs regulated by one or two of the agonists were identified . One of the identified miRNAs , miR - 190 , was down-regulated by fentanyl but not by morphine . This down-regulation was attenuated by 1,4- diamino -2,3- dicyano -1,4- bis ( methylthio ) butadiene ( U0126 ) , which blocks the phosphorylation of P29323 REA . When fentanyl-induced but not morphine-induced P29323 REA phosphorylation was blocked in the primary cultures from beta-arrestin 2 ( - / - ) mouse , fentanyl did not decrease the expression of miR - 190 . However , a PKC inhibitor that blocked morphine-induced P29323 REA phosphorylation specifically had no effect on the miR - 190 down-regulation . Therefore the decrease in miR - 190 expression resulted from the agonist-selective P29323 REA phosphorylation . In addition , the expressional changes in one of the miR - 190 targets , neurogenic differentiation 1 ( Q13562 REA ) , correlated with those in miR - 190 expression , suggesting the P35372 REA could regulate the Q13562 REA pathways via the control of miR - 190 expression .

Xaliproden ( SR57746A ) induces P08908 REA receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 REA agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 REA and P28482 REA isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 REA adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT - 1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 REA antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 REA stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 REA . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 REA receptors , P38936 REA Ras and MEK - 1 and by PKC and Akt pathways .

Anti-stress effect of astragaloside IV in immobilized mice . ETHNOPHARMACOLOGICAL RELEVANCE : Astragaloside IV , a major component extracted from the roots of Astragalus membranaceus ( AM ) , possesses anti-inflammatory , anti-oxidative , anti-fibrotic , anti-infarction and immunoregulatory effects . To clarify anti-stress effect of AM , anxiolytic and anti-inflammatory effects of 80 % ethanol extract of AM and astragaloside IV were investigated in immobilization stress model . MATERIALS AND METHODS : The mice were orally administered with AM ( 50 , 200 , and 500 mg / kg ) , astragaloside IV ( 5 , 10 , and 20 mg / kg ) and buspirone , a positive drug , 1h before immobilization treated for 2h . For anxiolytic activity assay , EPM test was performed in mice . For anti-inflammatory activity assay , serum levels of corticosterone , P05231 REA and P01375 REA - α were measured using ELISA kits . RESULTS : AM extract and astragaloside IV increased dose-dependently time spent on open arms and open arm entries in the EPM test . Anxiolytic effects of AM extract ( 500 mg / kg ) and astragaloside IV ( 20 mg / kg ) were comparable to those of buspirone ( 1 mg / kg ) . Their anxiolytic effects were blocked by WAY - 100635 ( 0.5 mg / kg , i . p . ) , a P08908 REA receptor antagonist ( p < 0.01 ) , but not by flumazenil ( 3 mg / kg , i . p . ) and bicuculline ( 0.5 mg / kg , i . p . ) , GABAA receptor antagonists . AM extract and astragaloside IV also reduced serum levels of corticosterone , P05231 REA and P01375 REA - α dose-dependently . CONCLUSIONS : AM , particularly astragaloside IV , may ameliorate immobilized stress-induced anxiety and inflammation .

Statin Modulation of Human T-Cell Proliferation , IL - 1β and Q16552 REA Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 REA inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD 3/28- stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 REA ( + ) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 MEN and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 REA production ( P < 0.01 ) . However , in response to anti-CD 3/28 stimulation , simvastatin significantly upregulated IL - 1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD 3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells .